Chinese Herbal Medicines -->Original articles

Identification of anti-inflammatory components in Panax ginseng of Sijunzi Decoction based on spectrum-effect relationship

2023/1/30 8:18:48

Objective: This study aimed to identify the main medicinal active components of Panax ginseng (P. ginseng) in the compatibility environment of clinical application. For this purpose, the anti-inflammatory ingredients of P. ginseng were investigated based on its therapeutic effect in Sijunzi Decoction (SJD) which is a widely used traditional Chinese formula. Methods: The fingerprints of 10 batches of SJD consisting of different sources of P. ginseng were established by UPLC technique to investigate the chemical components. At the same time, the anti-inflammatory effects of these components were evaluated by dextran sulfate sodium-induced ulcerative colitis mouse model. Grey relational analysis was applied to explore the correlation degree between fingerprints and anti-inflammatory effects in SJD. Lipopolysaccharide-stimulated RAW264.7 murine macrophages were established to evaluate the anti-inflammatory action of the screened effective substances of P. ginseng. Results: According to grey relational analysis, notoginsenoside R1, ginsenoside Rg2 and ginsenoside Rb3 of P. ginseng were the major anti-inflammatory contributions in SJD. They had been proven to be closely associated with the anti-inflammatory process of SJD and displayed a close effect compared with SJD by LPS-stimulated RAW264.7 murine macrophages. Conclusion: Our work provides a general strategy for exploring the pharmacological ingredients of P. ginseng in traditional Chinese formulas which is beneficial for establishing the quality standards of traditional herbs in traditional Chinese medicine prescription based on their clinical therapeutic effect.

Oligomeric Procyanidins Induce Generation of Reactive Oxygen Species and Collapse of Mitochchondrial Membrane Potential in Glioblastoma Cell Lines

Objective The aim of the present study was to clarify the mechanism underlying glioma cell death upon oligomeric procyanidins (F2) exposure. Methods The cytotoxicity of F2 on U87 (human malignant glioblastoma cell line) and C6 (rat glioma cell line) cancer cells was evaluated, and changes of mitochondrial membrane potential (MMP) and production of reactive oxygen species (ROS) in drug-treated cells were monitored. Moreover, morphological changes associated with F2-induced cells death were examined. Results F2 induced a concentration-dependent increase in ROS production and decrease in MMP. Furthermore, pre-incubation with N-acetylcysteine (NAC) and rotenone (Rt), resulted in partial inhibition of F2-induced ROS generation and marked attenuation of cell death and the cytoplasmic vacuolization induced by F2. In addition, pretreatment with Rt markedly attenuated the MMP loss in F2-treated cells. However, pretreatment with NAC only markedly attenuated the MMP loss in F2-treated C6 cells. Conclusion The increase in ROS level is at least one of mechanisms associated with F2-induced glioma cell death as well as the cytoplasmic vacuolization formation that contribute to the cytotoxicity of F2 in glioma cells.

Studies on Absorption and Tansport of Limoninoids from Fructus Evodiae in Caco-2 Cell Monolayer Model

YANG Xiu-wei,TENG Jie,ZHAO Bo and ZHANG Lian-xue

Objective To study the intestinal absorption and transepithelial transport of three limoninoids: evodol (EVO), limonin (LIM), and shihulimonin A (SHIA), isolated from Fructus Evodiae [the unripe fruit of Evodia rutaecarpa and Evodia rutaecarpa var. bodinieri] in the human intestine. Methods The in vitro cultured human colon carcinoma cell line, Caco-2 cell monolayer model, was applied to studying the absorption and transepithelial transport of the three limoninoids from apical (AP) to basolateral (BL) side and from BL to AP side. The three limoninoids were measured by reversed-phase high performance liquid chromatography coupled with ultraviolet absorption detector. Transport parameters and apparent permeability coefficients (Papp) were then calculated and compared with those of Propranolol as a control substance of high permeability and Atenolol as a control substance of poor permeability. Results The Papp value of EVO and LIM from AP to BL side for absorption and transport were 1.78 × 10-5 cm/s and 1.16 × 10-5 cm/s, respectively, which was comparable to that of Propranolol with Papp 2.18 × 10-5 cm/s. Conclusion The absorption and transport of both EVO and LIM are main passive diffusion as the dominating process in Caco-2 cell monolayer model, and they were estimated to be high absorbed compounds. SHIA in Caco-2 cell monolayer model may be involved in metabolism in the transport processes.

Excretion of Four Catechins in Tea Polyphenols in Rats

LI Qiu-sha,ZOU Ling-li,FU Ting,HAN Guo-zhu,Lü Li,ZHOU Qin,LIN Ya-fang and LI Nan

Objective To investigate excretion profiles of the four major anti-oxidant active catechins, (–) epigallo- catechin-3-gallate (EGCG), (–) epicatechin-3-gallate (ECG), (–) epigallocatechin (EGC), and epicatechin (EC) in tea polyphenols (TP) in rats in order to provide experimental data for clinical uses and development of TP as a novel drug. Methods The above four catechins in urine, bile, and feces were simultaneously determined by high performance liquid chromatography coupled with ultraviolet absorption detector (HPLC-UV) assay with a binary gradient elution. The samples were extracted by ethyl acetate prior to HPLC. The quantification was carried out by peak area internal standard method. Following iv dosing TP 100 mg/kg to rats, the samples were collected at different time intervals up to 8 h (urine and bile) and 24 h (feces). Results The urinary Ae, 0-8 h (cumulative excretion amount over 8 h) of EGCG, ECG, EGC, and EC were, on the average, 150.83, 30.75, 116.69, and 254.56 μg, corresponding to fe, 0-8 h (cumulative excretion fraction of dose over 8 h) of 1.45%, 0.84%, 7.88%, and 10.73%, respectively; the biliary Ae, 0-8 h were 12.61, 42.64, 6.61, and 1.24 μg, corresponding to the fe, 0-8 h of 0.12%, 1.16%, 0.45%, and 0.053% , respectively. For fecal excretion, only EGCG and EGC were detected with Ae, 0-24 h of 7.38 μg (fe, 0-24 h of 0.07%) and 157 μg (fe, 0-24 h of 9.99 %), respectively. The fe, total (the total fe of 3 excretory routes) were 18.32%, 10.78%, 2.00%, and 1.64% for EGC, EC, ECG, and EGCG, respectively. Conclusion EGCG and EC are mainly excreted in urine, ECG in bile, and EGC in feces by reference to their Ae and fe. The excretion of the four catechins based on fe, total is ranked in order of EGC > EC > ECG > EGCG. Only small amount of four catechins are recovered in urine, bile, and feces, indicating an extensive metabolic conversion of catechins in the rat body.

Chemical Constituents from Fruits of

HAN Li and FU Hong-zheng

Objective To study chemical constituents from fruits of Gymnocladus chinensis. Methods The compounds were isolated by various chromatographic techniques and their structures were elucidated on the basis of spectral analyses. Results Ten compounds were isolated from the EtOAc extract and were identified as 2-methyl pentyl (2E, 6S)-6-hydroxy-2, 6-dimethyl-2, 7-octadienoate (1), (2E, 6S)-6-?-L-arabino pyranosyloxy-2, 6-dimethyl-2, 7- octadienoic acid (2), ethyl (2E, 6S)-6-hydroxy-2, 6-dimethyl-2, 7-octadienoate (3), 5, 2?, 5?-trihydroxy-3, 7, 4?- trimethoxyflavone (4), (2R, 3R)-3, 3?, 5, 5?, 7-pentahydroxyflavanone (5), 2?, 4?-dihydroxy-4-methoxychalcone (6), kaempferol (7), apigenin (8), 4?, 7-dihydroxyflavone (9), and 5, 7, 4?-trihydroxy-3?-methoxyflavone (10). Conclusion Compound 1 is a new compound, compound 3 is firstly obtained from natural source, and compounds 4 – 10 are isolated from G. chinensis for the first time. Furthermore, the assignments of 13C-NMR data of compounds 3 and 4 are reported in this paper for the first time.

A Specific and Sensitive LC-MS/MS Method for Simultaneous Determination of Four Major Active Catechins of Tea Polyphenols in Rat Plasma and Its Application to Pharmacokinetic Study

WANG Chang-yuan,LI Qiu-sha,HAN Guo-zhu,ZOU Ling-li,LV Li,ZHOU Qin and LI Nan

Objective To develop a liquid chromatography technique coupled with tandem mass spectrometry (LC-MS/MS) for simultaneous determination of four active catechins EGCG, ECG, EGC, and EC of tea polyphenols (TP) in rat plasma in order to further study its multi-component pharmacokinetics. Methods Following a single step liquid-liquid extraction of plasma samples with ethyl acetate, the four catechins were separated on a Hypersil ODS C18 column using an isocratic mobile phase composed of methanol-water (30︰70). The detection using a mass spectrometer was performed under negative ESI in the MRM mode. The analytes were identified by reference to both MRM and tR values and quantified using peak area internal standard method. Results The method was shown to be specific without interference from matrix, metabolites, and impurities present in TP raw material and to be sensitive with LOD and LOQ of 1.5 and 10 ng/mL (EGCG) as well as 0.75 and 5 ng/mL (ECG, EGC, and EC). A good linearity was obtained over a wide range of 10-10 000 ng/mL for EGCG and 5-5000 ng/mL for other three catechins (r > 0.996). The method was validated to be reproducible and reliable, as evidenced by intra-batch and inter-batch precision of less than 10% and 11%, accuracy of 97.13%-106.05% and 99.22%-103.14%, respectively. The recovery of extraction ranged from 72.74% to 89.13%, matrix effect from 88.76% to 105.97% for four catechins. The method was successfully used to study the pharmacokinetics of TP iv administered to rats at a dose of 100 mg/kg. Conclusion This method is shown to completely meet requirements for the multi-component pharmacokinetic study of TP in rats.

The Anti-inflammatory Effects of Components in Shuxiong Tablet and Its Possible Formulary Rationality

WEI Wei,XIE Sheng-rong,FU Ji-hua,LIU Wen,QI Jian-ping and PING Qi-neng

Objective To determine whether the anti-inflammatory properties of Shuxiong Tablet (SXT) and the effective components group of SXT (ECGS) are equivalent and to assess the formulary rationality. Methods ECGS consisted of Panax notoginsen saponion (PNS), hydroxysafflor yellow A, and ferulic acid plus volatile oil of Ligusticum chuanxiong, which was based on the active ingredients and their ratios in SXT. We compared the anti-inflammatory actions of ECGS and SXT using the xylene-induced edema model and the carrageenan-induced edema model, as well as the analgesic activity of them using the acetic acid-induced writhing model. Moreover, cultured macrophages were incubated with media containing serum isolated from SXT-, ECGS-, or every component of ECGS-treated rats, to compare the depress effects on lipopolysaccharide (LPS)-stimulated NO production and inducible nitric oxide synthase (iNOS) expression. Results ECGS and SXT had equivalent anti-inflammatory actions and analgesic effects at an equipotent dosage in a dose-dependent manner. The drug-containing media could inhibit the LPS-stimulated NO production and iNOS expression in cultured macrophages. A 2×2×2 ANOVA revealed that three effective components could produce synergistic effect on the inhibition of NO production, and PNS was the capital component. Conclusion ECGS and SXT display an equivalent anti-inflammatory effect, and the formula follows traditional Chinese medicine compatibility principle, which shows obvious formulary rationality.

Morphological and Chemical Variation of Prunella vulgaris Populations from Different Locations in China

LIAO Li,LIU Li,GUO Qiao-sheng,WANG Zhi-yong and CHEN Yu-hang

Objective To investigate the variation of chemical characteristics with environment and establish a relationship between morphological characters and chemical composition of Prunella vulgaris collected in different areas of China. Methods Twelve phenotypic traits and three chemical compositions were assessed in 28 populations of P. vulgaris collected from different locations in China. Results The variability ranges observed at phenotypic and chemical levels were polymorphic. According to the morphological traits, 28 populations of P. vulgaris could be grouped into six clusters, and two morpho-types could be clearly distinguished. Perceptible differences could be discerned in the plant height, leaf length, corolla length, calyx length, fruiting spikes length, and maturity period. Based on three kinds of components including ursolic acid, total flavonoids, and total polysaccharides, all populations could be identified as four types. Cluster IV showing high content of ursolic acid, total flavonoids, and total polysaccharides could be utilized to develop superior derivatives. Conclusion The variation of chemical characteristics is influenced by the genetic and environmental factors, such as soils, climates, longitudes, and altitudes. It provides a solid basis for efficiently evaluating qualities and establishing good agricultural practices for P. vulgaris.

Control of Stem Blight of Schizonepeta tenuifolia Caused by Phytophthora nicotianae Using Trichoderma spp.

LI Yong and DING Wan-long

Objective To control stem blight disease of Schizonepeta tenuifolia caused by Phytophthora nicotianae. Methods The antagonist effect of 13 Trichoderma strains (including T. viride and T. harzianum) was evaluated upon mycelia growth of P. nicotianae. Trichoderma strains with high antagonistic activities against the pathogen were used to control stem blight of S. tenuifolia in the field. Results Of 13 Trichoderma strains tested, T. viride strain M3 showed maximum mycelia growth inhibition (83.2%) to the pathogen, followed by T. viride strain Tv04-2 (78.2%) and then T. harzianum strain ThB (65.0%), in vitro. Fungal cell wall degrading enzymes, protease, and β-1,3-glucanase were analyzed qualitatively and quantitatively in further study. T. viride strains M3, Tv04-2, and T. harzianum strain ThB efficiently against P. nicotianae were used to control stem blight of S. tenuifolia in the field, and T. viride strain M3 showed the best biocontrol potential. Conclusion Trichoderma spp. can be used as alternatives of pesticides to control stem blight, one of the serious soilborne diseases of S. tenuifolia caused by P. nicotianae. However, though T. viride strains Tv04-2 and T. harzianum strain ThB are also highly against P. nicotianae in vitro, the controlling efficacy of them on stem blight disease is not as excellent as T. viride strains M3 in the field.

A New Triterpene Saponin from Gynostemma pentaphyllum

SHI Lin,CAO Jia-qing,ZHAO Hong and ZHAO Yu-qing

Objective To study the triterpene saponins from Gynostemma pentaphyllum with antitumor activities. Methods The 75% EtOH extract of G. pentaphyllum was used for isolation by silica gel column chromatography and preparative HPLC. The structures of pure compounds isolated were identified by the spectral analysis and chemical evidence. Results Two compounds were isolated and identified as 23(S)-3β,20ξ,21ξ-trihydroxy-19-oxo-21,23- epoxydammar-24-ene 3-O-α-L-rhamnopyranosyl (1→2)-[β-D-xylopyranosyl (1→3)]-β-D-arabinopyranoside (1) and 23(S)-21(R)-O-n-butyl-3β,20ξ-dihydroxy-21,23-epoxydammar-24-ene 3-O-α-L-rhamnopyranosyl (1→2)-[β-D- xylopyranosyl(1→3)]-β-D-arabinopyranoside (2). Conclusion Compound 2 is a new triterpene saponin with moderate antitumor activities against the HL-60, Colon205, and Du145 cell lines.

Optimization of Microwave-assisted Extraction of Polyphenols from Enteromorpha prolifra by Orthogonal Test

LUO Hong-yu,WANG Bin,YU Chun-guang and XU Yin-feng

Objective To optimize microwave-assisted extraction of polyphenols from Enteromorpha prolifra. Methods Based on single-factor tests, an efficient microwave-assisted extraction (MAE) technique was developed to extract bioactive polyphenols from E. prolifra through orthogonal (L16(4)5) test. Results The highest yield (0.923 ± 0.013) mg/g was obtained when microwave power, solvent to raw material ratio, irradiation time, ethanol concentration, and extraction cycles were 500 W, 25 mL/g, 25 min, 40%, and 3, respectively, which was higher than that of Soxhlet extraction with methanol for 6 h, ultrasound-assisted extraction with 40% ethanol for 1 h twice and heat reflux extraction with 40% ethanol for 2 h twice. Conclusion This finding indicates that MAE is a superior technique for the extraction of polyphenols due to less impurity, higher time efficiency and yield.

Inhibition of Emodin on LPS-induced Nitric Oxide Generation by Suppressing PLC-γ Phosphorylation in Rat Peritoneal Macrophages

WANG Xin-yu,CAI Shou-guang,WU Yi-fen,LI Jun-ying,YANG Wen-xiu and HU Fen

Objective To investigate the inhibitory mechanism of emodin on lipopolysaccharide (LPS)-induced nitric oxide (NO) generation in rat peritoneal macrophages. Methods NO production and iNOS expression were measured through nitrite assay and Western blotting assay, respectively. NF-κB activity and nuclei P65 expression were estimated by dual-luciferase and Western blotting assay, respectively. Intracellular free Ca2+ ([Ca2+]i) was detected using the ratiometric fluorescent calcium indicator dye, Fura-2, and a microspectrofluorometer. PLC-γ phosporylation was analyzed by Western blotting assay. Results First, emodin was found playing active roles in suppressing LPS-induced NF-κB activation in rat peritoneal macrophages. Second, emodin down-regulated transient [Ca2+]i and could increase in NF-κB upstream signal. Finally, emodin suppressed phosphorylation of PLC-γ by LPS stimulation in the upstream of [Ca2+]i. Conclusion Suppression of PLC-γ phosphorylation is involved in emodin inhibiting NO generation by LPS stimulation in rat peritoneal macrophages.

Enhancement of Extracts from Celastrus orbiculatus on Maturation and Function of Dendritic Cells in vitro and in vivo

QIAN Ya-yun,ZHANG Hua,YUAN Lin,HOU Ying,LIU Wei-wei and LIU Yan-qing

Objective To examine the immunoregulation of Celastrus orbiculatus extracts (COE), a traditional Chinese medicine, on maturation and function of dendritic cells (DCs) in vitro and in vivo. Methods In vitro, after treated with COE in different nontoxic concentrations (0, 10, 20, 40, 80, and 160 μg/mL) for 5 d, the surface immunological molecules and cytokine secretion of mice bone marrow-derived DCs in response to COE were analyzed by flow cytometric analysis (FACS) and enzyme linked immunosorbent assay (ELISA), respectively. In vivo, mouse hepatoma cells (Hepa1-6, 1 × 106) were injected sc and were treated with different dosages of COE (10, 20 or 40 mg/kg/d). Effects on tumor growth were determined by tumor volume and histology analysis after 28 d administration of COE. The relative proportions of mature DCs and CD8+ T cells were measured in mononuclear cells that had been isolated from spleen by FACS. Results COE stimulated IL-2 and IFN-γ secretion of DCs, simultaneously enhanced the maturation of DCs by enhancing immunological molecule (CD40, CD80, CD86, H-2Kb, and I-Ab) expression in a dose-dependent manner. Furthermore, the chemotactic responses of DCs were significantly higher in COE-treated than untreated DCs, in association with higher chemokine receptor 7 expression. Furthermore, COE increased DCs produce IFN-γ and IL-2 in a dose-dependent manner when the concentration of COE less than 40 μg/mL, decreased DCs produce IL-10 and IL-4 also in a dose-dependent manner. In in vivo studies, COE can not only suppress growth of malignant hepatocellular carcinomas but also stimulate maturation of DCs, associated with strongly enhanced CD8+ CTL responses. Conclusion These data provide new insight into the mechanism of action of COE and indicate that the stimulation of maturation and function of DCs by COE contributes to its immunoregulatory effects.

Anti-oxidation of Tanshinone ⅡA and Prohibitin on Cardiomyocytes

YANG Ping,JIA Yu-hua,LI Jie,ZHOU Feng-hua and LI Li-jun

Objective To investigate the anti-apoptotic mechanism of tanshinone ⅡA and the function of prohibitin (PHB) on myocardial cells apoptosis induced by hydrogen peroxide (H2O2). Methods Myocardial cells were primary cultured neonate rat were cultured in medium with 200 μmol/L H2O2, and the medium was supplemented with tanshinone ⅡA (1 × 10－4 mol/L) in advance for 24 h. PHB in myocardial cells was knocked down by RNA interference, and the expression level of PHB was determined by Western blotting analysis. Flow cytometric analysis was used to detect apoptosis rate, intracellular calcium concentration ([Ca2+]i), and mitochondrial membrane potential (MMP). Results H2O2-mediated cell apoptosis resulted in activation of PHB, increasing of [Ca2+]i, and decreasing of MMP. Tanshinone ⅡA profoundly inhibited myocardial cell apoptosis induced by H2O2, decreased [Ca2+]i, and increased MMP. Speci?c silence of PHB by siRNA down-regulated the expression level of PHB, increased apoptosis rate and [Ca2+]i, and decreased MMP. Conclusion The results demonstrate that tanshinone ⅡA could attenuate apoptosis induced by H2O2, and the activation of PHB induced by H2O2 is the major regulatory pathway of cyto-protective gene expression against oxidative stress.

Different Effects of Mahuang Decoction and Maxing Shigan Decoction on Animal Temperature Tropism and Correlation to Differences of Cold and Hot Nature of Chinese Materia Medica

Objective To establish an objective method for evaluating the intrinsic characteristics between cold and hot nature of Chinese materia medica (CMM) through the different effects of Mahuang decoction (MHD) and Maxing Shigan decoction (MSD) on animal temperature tropism. Methods The equipment with cold/hot pads was used to investigate the variety of the temperature tropism between two groups of mice treated by MHD and MSD, respectively. Meanwhile, the activities of adenosine triphosphatase (ATPase), superoxide dismutase, succinate dehydrogenase, and malondialdehyde were measured. Results After treated by MHD, the macroscopic behavioral index of remaining rate on warm pad (40 ℃) of mice decreased significantly (P < 0.05), suggesting the enhancement of cold tropism, meanwhile, the internal indices of ATPase activity and oxygen consuming volume increased significantly (P < 0.05), suggesting the enhancement of energy metabolism. On the other hand, the above-mentioned indices in MSD group changed on the inverse way. Conclusion The relative drug nature of MHD and MSD revealed in this study is consistent with the theoretical prognostication or definition. It indicates that the internal cold and hot nature of CMM could be reflected in ethological way on the changes of animal temperature tropism which might be internally regulated by body energy metabolism.

Quality Evaluation and Regional Analysis of Psoraleae Fructus by HPLC-DAD-MS/MS plus Chemometrics

YAN Dong-mei,CHANG Yan-xu,KANG Li-yuan and GAO Xiu-mei

Objective To evaluate the quality of Psoralea corylifolia collected from 12 provinces of China. Methods An HPLC-DAD-MS/MS method was used to identify, determine, and estimate 14 representative bioactive compounds in P. corylifolia. Then on the basis of the content data, the chemometrics method was used to differentiate 20 samples from different regions. Results The quality of P. corylifolia from 12 different provinces of China was evaluated by this method. Though the samples showed similar profiles, content of the detected markers varied significantly in different regions and batches. According to the results of the hierarchical cluster analysis and principle component analysis, it can be concluded that the samples from different origins could be clustered reasonably into two groups, as well as successfully distinguished. Conclusion A simple and reliable new method which used HPLC-DAD-MS/MS and chemometrics has been developed to characterize, classify, and control the quality of P. corylifolia.

Plant Regeneration through Somatic Embryo in Herpetospermum pedunculosum, an Endangered Tibetan Medicinal Herb

Objective An effective reproducible protocol for complete plant regeneration via somatic embryogenesis has been developed for Herpetospermum pedunculosum, an endangered Tibetan medicinal herb. Methods The cotyledon explants used in this study were excised from seedlings germinated in vitro. Callus was induced from cotyledon explants on Murashige and Skoog’s medium, supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 0.1–1.0 mg/L) alone or in combination with 6-benzylaminopurine (BA, 0.5, 1.0, and 2.0 mg/L). Results The calli showed differentiation of globular embryos after three weeks of incubation on MS medium supplemented with various combinations of BA and NAA. Sixty-two percent of the embryogenic calli produced somatic embryos in MS basal medium supplemented with BA (1.0 mg/L) + NAA (2.0 mg/L). The addition of KN (0.5 mg/L) to MS medium containing both BA and NAA (2.0 mg/L each) significantly increased the frequency of somatic embryogenesis. The maximum percentage of embryogenic calli formation was 83%, and globular embryos formed and germinated successfully in this medium. Then, transferring the regenerated plants from this medium to hormone-free MS medium will further enhanced the development of the plants, and the healthy plantlets are formed successfully within four weeks. The plantlets were transferred to soil to acclimatize under greenhouse conditions and 75% survived. Conclusion Somatic embryogenesis protocol as reported here can play a key role in the propagation and conservation of this endangered species.

Anti-inflammatory and Analgesic Effects of Extract from Roots and Leaves of Citrullus lanatus

DENG Jia-gang,WANG Shuo,GUO Li-cheng and FAN Li-li

Objective To study anti-inflammatory and analgesic effects of extract from the roots and leaves of Citrullus lanatus and assess their acute toxicity in animals. Methods The mouse model with ear edema induced by xylene and the rat model with paw edema or granuloma by carrageenin or cotton pellet were used for anti-inflammatory effects of the extract. Effects of the extract on analgesia was tested respectively by measuring the latency of mice licking hind foot from hot plates and by counting the times of body twisting in response to acetic acid. The acute toxicity of the extract was determined with the method of Bliss. Results The extract significantly inhibited the ear edema, granuloma hyperplasia, and paw edema. It significantly lifted the pain threshold on mouse hot-plate responses and reduced their writhing times. During the 7 d observation period in its acute toxicity assay, no apparent toxic reaction was shown and all mice survived at a dose of 87 g extract per kg body weight. Conclusion The extract could protecte mice/rates from inflammation and analgesia, and may be safe as an orally administered natural product for humans.

Virtual Evaluation on the Activities of Phthalides and Terpenoids from Angelica sinensis

Objective To elucidate potential activities of phthalides and terpenoids from Angelica sinensis by theoretical docking calculation. Methods Eleven components of phthalides and terpenoids were selected as ligand. The crystalline structures of targets related to common diseases were used as the receptors for calculation. The calculations were conducted with Schr?dinger software package. The grading standard of selectivity was developed according to G-score between ligands and receptors. Results Selective targets of phthalides and terpenoids were related to nevous system diseases, cancer, pain, diabetes, cardiovascular disease, liver cirrhosis, nephrotic syndrome, inflammatory diseases, rheumatoid arthritis, dermatosis, leukemia, microbial inflections, immune diseases, and hypercholesterolemia. In addition to the medical treatments reported in the literature, our research also indicated that these two classes of compounds may be used for tumor, diabetes, rheumatoid arthritis, dermatosis, leukemia, liver cirrhosis, and nephrotic syndrome. According to our research, the effects of phthalides and terpenoids may be not so strong. Conclusion The effects of phthalides and terpenoids on diseases founded through virtual evaluation accord greatly with those reported in experiment and clinic. The combination of computer-aided drug evaluation technique and experiment is definitely an important and fast way to investigate the effects and mechanisms of traditional Chinese medicine.

Simultaneous Analysis of 13 Major Bioactive Components in Evodiae rutaecarpa by HPLC-DAD-MS

XU Hai-yu,ZHANG Tie-jun,XIAO Xue-feng,ZHAO Ping,LIU Chang-xiao and XU Jun

Objective To control the quality of Evodia rutaecarpa better. Methods An HPLC-DAD-MS/MS method was established for the rapid and efficient identification of bioactive constituents and for simultaneous quantitative analysis of four bioactive ingredients including evodiamine, rutaecarpine, dehydroevodiamine, and evodin in E. rutaecarpa, which was applied to evaluating eight samples of E. rutaecarpa and its varieties from different areas. Results Thirteen potentially bioactive constituents including one flavonoid glycoside, one limonin, four indoloquinazoline alkaloids, and seven quinolone alkaloids were identified in all samples and the contents of dehydroevodiamine, evodine, evodiamine, and rutaecarpine varied widely from 0.10% to 0.51%, 0.49% to 3.12%, 0.07% to 1.56%, and 0.10% to 0.69%, respectively. Conclusion This method is found to be convenient, fast, accurate, and it is facilitated to improve the quality control standard of E. rutaecarpa and related products.

Prevention of Combination of Hirsntella sinensis and Panax notoginseng Extracts on Bleomycin-induced Pulmonary Fibrosis in Mice

Objective Hirsntella sinensis (HS) is the anmorph of Ophiocordyceps sinensis (Cordyceps sinensis). O. sinensis and Panax notoginseng are two popular Chinese herbs, commonly used in traditional Chinese prescriptions for the treatment of various diseases. A combination of HS extract with P. notoginseng saponin (PNS) extract demonstrated more prominent lung-protective activity than the two herbs individually used in our preliminary studies. This study further investigated the action of their combination (HSPNS) on anti pulmonary fibrosis using a Bleomycin (BLM) induced mouse model. Methods BLM-treated Kunming mice was given HSPNS daily for 7, 14 or 28 d via ig administration. After treatment, following parameters were monitored using proper methods, respectively. Lung index, serum and lung malondialdehyde (MDA) and hydroxyproline (HYP) contents, lung superoxide dismutase (SOD) activity, transforming growth factor β1 (TGF-β1), and expression levels of collagen Ⅰ (Col-Ⅰ) and collagen Ⅲ (Col-Ⅲ). The lung biopsies were also dissected for semiquantitative histological analysis. Results The results indicated that HSPNS significantly reduced lung index, MDA and HYP contents, and expression levels of TGF-β1, Col-Ⅰ, and Col-Ⅲ. The combination also remarkably enhanced SOD activity compared with BLM-induced group. Moreover, the severe pulmonary fibrosis histopathological changes induced by BLM could be attenuated by HSPNS treatment. Conclusion These results suggest that HSPNS could significantly inhibit the progression of pulmonary fibrosis induced by BLM and its inhibitory effect might associate with its ability to scavenge free radicals, decrease TGF-β1 level, and inhibit collagen synthesis.

Neuroprotection of Glycyrrhizin against Ischemic Vascular Dementia in vivo and Glutamate-induced Damage in vitro

HOU Yuan-yuan,YANG Yang,YAO Yang and BAI Gang

Objective To evaluate the neuroprotective effects of glycyrrhizin (GL) on vascular dementia (VaD) in vivo and Glutamate-induced damage in vitro. Methods Male Wistar rats were subjected to permanent occlusion of the bilateral common carotid arteries. On d 15 postsurgery, rat cognition was assessed using the Morris water maze. The activity of superoxide dismutase and the content of malondialdehyde of brains were also measured. Brain damage was evaluated histologically using HE staining. In vitro cell viability was examined in PC12 cells exposed to Glutamate, and mRNA levels of Bcl-2 and Bax were assessed. Results GL (20 mg/kg for 12 d) improved the performance of learning and memory of VaD rats, decreased the level of lipid peroxidation, and attenuated the pathological alterations in the hippocampal CA1 and CA3 areas. Moreover, GL (0.6 mmol/L) could protect PC12 cell lines from injury induced by Glutamate (10 mmol/L) and inhibit apoptosis of neuronal cells. Conclusion The present findings suggest that GL may have therapeutic potential in treating VaD.

Determination of Shionone in Rat Plasma by HPLC and its Pharmacokinetic study

TIAN Ya-ping,WANG Qiao,YANG Wei,KONG De-zhi and ZHANG Lan-tong

Objective To develop a sensitive, simple, and accurate method for the determination of shionone in rat plasma after ig administration of Asteris Radix petroleum ether extract (RAPE). Methods The separation was achieved by HPLC on a RP18 column (150 mm × 3.9 mm, 5 μm) with a mobile phase composed of acetonitrile-0.05% phosphoric acid water (98︰2) at a flow rate of 1.0 mL/min. UV Detector was set at 200 nm and friedelin was chosen as an internal standard. Results The linear range of the standard curves was (0.3443–22.0) μg/mL with the correlation coefficient of 0.9968. The intra- and inter-day precisions were all below 10% and the relative error was ?3.5%–1.1%. Conclusion The developed method can be successfully applied to the pharmacokinetic study. After ig administration of RAPE, T1/2(ka) is (33.09 ± 7.32) min and T1/2(ke) is (84.95 ± 22.34) min.

Virtual Evaluation on Activities of Flavonoids from Scutellaria baicalensis

Objective To explore the investigation method of complicated and profound traditional Chinese herbal medicine, the potential action mechanisms of flavonoids from Scutellaria baicalensis were studied by docking calculation. Methods In total, eight flavonoids (aglycones and their glicosides) from S. baicalensis were selected as ligands. The crystalline structures of targets related to common diseases were used as the receptors for calculation. The calculations were conducted with Schr?dinger software package. The grading standard of selectivity was developed according to G-score between ligands and receptors. Results Twenty-six pharmacologic actions have been reported. Among all effects in literature, nine of them can be deduced from the docking calculation of aglycone. From glycosides with grade ++, 25 reported effects can be estimated by calculation. Apparently, the target selectivity of aglycones and their glycosides are different form the virtual evaluation. The virtual evaluation results of glycosides were closer to the reported effects. Conclusion Our proposed virtual evaluation method seems an effective way to investigate the complicated system of traditional Chinese herbal medicine. It suggests that aglycones may be effective as the form of glucoside in vivo, and metabolism is a very important factor for virtual evaluation.

Derivative Synthesis of Wanpeinine A, a Major Steroidal Alkaloid from Fritillaria shuchengensis

WEN Juan,ZHOU Xian-li,YAN Ren-long,HUANG Shuai and WANG Yin-hui

Objective To design and synthesize derivatives of wanpeinine A, the main steroidal alkaloid isolated from the plant Fritillaria shuchengensis, and further study on the structure-activity relationship of the steroidal alkaloid. Methods Acylation and alkylation were used to synthesize the derivatives and their structures were identified via NMR and MS. Results The acylation of wanpeinine A (1) produced 3β,6α-diacetylwanpeinine A (2), 3β,6α-dipropionyl- wanpeinine A (3), 3β,6α-dichloracetylwanpeinine A (4), 3β,6α-dibenzoylwanpeinine A (5), and 3β-methoxy- acylwanpeinine A (6). The alkylation of wanpeinine A formed 3β,6α-dimethoxymethylwanpeinine A (7). Conclusion All compounds are new except for 3β,6α-diacetylwanpeinine A .

Triterpenoid Saponins from Roots of Gypsophila pacifica

NIE Wei,LUO Jian-guang and KONG Ling-yi

Objective To study the chemical constituents from the roots of Gypsophila pacifica. Methods The chemical constituents were isolated by various column chromatographic methods and their structures were identified by spectral data together with physicochemical analysis. Results Five compounds were isolated and identified as 3-O-β-D- galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl gypsogenin 28-O-β-D-xylopyranosyl- (1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranoside (1), 3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl- (1→3)]-β-D-glucuronopyranosyl quillaic acid 28-O-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D- fucopyranoside (2), 3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl gypsogenin 28-O-β-D-glucopyranosyl-(1→3)-[β-D-xylopyranosyl-(1→4)]-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranoside (3), 3-O-β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl quillaic acid 28-O-β-D- glucopyranosyl-(1→3)-[β-D-xylopyranosyl-(1→4)]-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranoside (4), 3-O-β-D- galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucuronopyranosyl quillaic acid 28-O-α-L-arabino- pyranosyl-(1→4)-α-L-arabinopyranosyl-(1→3)-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fuco-pyranoside (5). Conclusion The five compounds are isolated from this plant for the first time.

Isolation and Purification of Isoaloeresin D and Aloin from Aloe vera by High-speed Counter-current Chromatography

WAN Jin-zhi,CHEN Xin-xia,QIU Chun-mei and LIN Xiang-cheng

Objective To develop an efficient method to isolate and purify the main components isoaloeresin D and aloin from Aloe vera for its industrial production. Methods High-speed counter-current chromatography was used to isolate isoaloeresin D and aloin in a one-step separation from dried crude extract of A. vera. The biphasic solvent system composed of hexane-ethyl acetate-acetone-water (0.2︰5︰1.5︰5) was used at a flow rate of 1.0 mL/min, while the lipophilic phase was selected as the mobile phase and the apparatus was rotated at 840 r/min. The effluent was detected at 254 nm. Results Isoaloeresin D (53.1 mg) and aloin (106.9 mg) were separated from the crude extract (384.7 mg) with the purities of 98.6% and 99.5%, respectively. Conclusion HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.

Fingerprint Analysis of Four Variants of Chrysanthemi Morifoli Flos by RP-HPLC

SHEN Zhi,ZHANG Wen-ting,HUA Yun-fei and ZHAO Wei-liang

Objective To establish a RP-HPLC fingerprint analysis method for quality evaluation and control of the four variants of Chrysanthemi Morifoli Flos. Methods RP-HPLC was used to establish the fingerprint method. Results Despite of the similarity in terms of holistic HPLC chromatograms, the four variants of Chrysanthemi Morifoli Flos exhibit characteristic fingerprints and can be readily recognized by similarity clusters. Conclusion A simple and reliable HPLC fingerprint method has been developed and validated to authenticate the four variants of Chrysanthemi Morifoli Flos, providing a scientific basis for quality control of Chrysanthemi Morifoli Flos.

Relationship between DNA Barcoding and Chemical Classification of Salvia Medicinal Herbs

Objective To make the identification of medicinal herbs in Salvia L. quickly and accurately. Methods In this work, DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L. First, the nucleotide sequences of the internal transcribed spacer region two amplified from 48 medicinal plants in Salvia L., and three other groups of medicinal plants in Lamiaceae were sequenced. A molecular phylogeny was constructed using the minimum evolution and maximum parsimony methods according to their sequence diversity. Second, the water-solution bioactive components and lipid soluble components were tested by HPLC. Then a chemical phylogeny was built using HPLC fingerprint data. Comparing the molecular and chemical phylogenetic trees revealed many similarities. Results DNA barcoding was sequencing based and could therefore provide more accurate results within a shorter time especially in large-scale studies. Conclusion The results show that ITS2 region is a novel DNA barcode for the authentication of the species in Salvia L. This is the first work to show the relationship between DNA barcoding and chemical components.

Toxicological Assessment of Trans-resveratrol

ZENG Jian-guo,HU Yi-xiu and ZHANG Xiang-qian

Objective To evaluate toxicity and safety of trans-resveratrol (t-RSV). Methods For assays of acute toxicity, genetic toxicity, and sub-chronic toxicity, Ames test, mice bone marrow erythrocyte micronucleus, and mice sperm abnormality were performed. Results In the acute oral toxicity tests, maximum tolerable dose (15 g/kg) in male and female Kunming mice showed no toxicological signs. For 90-d feeding of t-RSV at dosage range of 167–500 mg/(kg?d) in both male and female Sprague-Dawley rats, no noticeable toxicological effects were observed. Conclusion T-RSV has no acute toxicity and no genotoxicity, no harmful effects on the human body at the tested dosage range and thus resveratrol is safe for human consumption.

Transformation of Compound K from Saponins in Leaves of Panax notoginseng by Immobilized β-Glucanase

DONG Hui-juan,JIANG Bin-hui,HAN Ying,GENG Yong and ZHAO Yu-qing

Objective To prepare an active anti-tumor component, compound K (C-K), from saponins in leaves of Panax notoginseng (SLPN) using immobilized β-glucanase. Methods Two entrapments, alginate gel-1 (Alg 1) and alginate gel-2 (Alg 2), were evaluated for their ability to immobilize β-glucanase. The amount and purity of C-K obtained from the transformation process were analyzed by HPLC, and the immobilizing parameters were optimized. Results β-Glucanase can be immobilized and reused with either of the entrapment. However, using Alg 1 resulted in higher enzyme activity than Alg 2. The optimal concentration of the immobilized enzyme was 10%; The optimal crosslinking time was 4–6 h; and the optimal concentration of the crosslinking agent was 6%–7%. Conclusion Immobilized β-glucanase shows sustained enzyme activity, good ethanol tolerance, and was reusable for the preparation of C-K from SLPN.

Effects of Astragaloside Ⅳ Derivative on Heart Failure in Rats

WANG Wei-ting,ZHAO Zhuan-you,HAN Ying-mei,XU Wei-ren and TANG Li-da

Objective Astragaloside Ⅳ derivative (ASId) is one of Astragaloside Ⅳ (ASI) derivatives with higher water-solubility and may have more druggability than ASI. The present study aims at observing the effects of ASId on cardiovascular parameters in chronic heart failure in rats. Methods Using echocardiographic and haemodynamic measurements, the effects of ASId on congestive heart failure (CHF) induced by ligation of the left coronary artery in rats were investigated. Results ASId iv 0.5, 1.0, and 2.0 mg/(kg?d) attenuated the decline of ejection fraction. The peak derivatives of the left ventricle (LV) pressure (dp/dt) in ASId treated groups were significantly increased. Both LV volumes in diastole and in systole were decreased significantly after ASId treatment, accompanied with a trend towards normalization of relative wall thickness at end-systole. ASId 0.5, 1.0, and 2.0 mg/(kg?d) attenuated the increase of LV systolic and diastolic wall stress. ASId treatment also inhibited compensatory hypertrophy of depressed heart. Conclusion ASId could improve cardiac functions and inhibite compensatory hypertrophy and LV remodelling, which suggests the possibility of ASId as a new therapeutic drug for the treatment of CHF.

Influence of Different Extraction and Purification Methods on Astragalus Polysaccharides and Pharmacological Evaluation

YANG Yi-fang,FENG Jing-qian,XU Hai-yan,PENG Dan-ming and HU Jian-ping

Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides (APS) as complementary therapeutic agents prepared by different extraction and purification techniques. Methods Components of APS prepared by different extraction and purification techniques were analyzed, and these APS were used for synergy and attenuation of chemotherapy, radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice, and also used for the regulation of immune function to immunosuppression mice. Results Experimental data were analyzed by means of statistical method to get pharmaco-result: A3 (extracted by microwave assistance and purified by membrane separation) > A4 (extracted by refluxing and purified by membrane separation) > A1 (extracted by refluxing and no purification) ≈ A2 (extracted by microwave assistance and no purification). There were no significant differences on pharmacodynamic action between A1 and A2. However, compared with A1 and A2, it was worth noting that A3 and A4 exhibited good pharmacodynamic action. Then A3-in and A4-in, the samples in dialyzer after dialysis, were separated and purified to get homogeneous APS, which were the principal constituents of APS in dialyzer, with the molecular weight (Mw) of 7669 and 14 142 determined by HPGPC, respectively. The average Mw of APS outside of the dialyzer, A3-out was 3102 and A4-out 3256, which were the main compositions of A3 and A4, accounted for 79.63% and 53.92%, respectively. Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity. Refluxing, microwave assistance extractions, and membrane enrichment techniques bring different cases on Mw distribution, components and pharmacodynamic action, and obviously exhibit relationship among component, Mw distribution, and pharmacological action.

A New Steroidal Glycoside from Roots of Cynanchum auriculatum

YIN Min,FENG Xu,CHEN Yu,WANG Ming,XIA Bing and DONG Yun-fa

Abstract: Objective To study the chemical constituents of Cynanchum auriculatum. Methods Two steroidal glycosides were purified by column chromatography and their structures were elucidated by spectroscopic analysis. Results and Conclusion Compound 1 is a new steroidal glycoside, named kidjoranin-3-O-β-D-cymaropyranoside. Compound 2 is isolated from this plant for the first time.

Chemical Constituents of Ganoderma resinaceum

DING Ping,QIU Jin-ying and LIANG Ying-jiao

Abstract: Objective To study the chemical constituents of Ganoderma resinaceum. Methods Chemical constituents were isolated and purified by silica gel column, Sephadex LH-20, and ODS column chromatography. The structures were identified by means of physicochemical and spectral data. Results From the CHCl3 part of the fruiting bodies of this material, eight compounds were isolated. Their structures were determined to be ergosterol (1), β-sitosterol (2), ganoderic acid E (3), ganoderic acid G (4), ganoderic acid C2 (5), ganoderic acid B (6), ganoderic acid A (7), and lucidenic acid A (8). Conclusion All the compounds were obtained from this fungus for the first time.

Regulation of Fengxiaofang on Expression of Gene Gob-5 in Asthmatic Mice

Abstract: Objective To observe the regulation of Fengxiaofang on the expression of gene Gob-5 in ovalbumin-challenged asthmatic mice. Methods The murine model of allergic asthma established by ovalbumin was used. The model mice were treated experimentally with Fengxiaofang and Dexamethasone as a positive control. The quantitive expression of gene Gob-5 in lung of mice was detected by real-time quantitative PCR. Results The expression of gene Gob-5 in lung of mice was down-regulated by Fengxiaofang. In the experiment, there were no significant differences between the one-fold or ten-fold dosage groups and the normal control group. Gene Gob-5 expression in five-fold dosage was significantly lower than that in the normal control group. Conclusion Fengxiaofang has the effect of lowering the expression of gene Gob-5 which maybe related to the potency on asthmatic disease.

Inhibition of Celastrus orbiculatus Extracts on VEGF Expression in Hepatoma Cells of Mice

QIAN Ya-yun and LIU Yan-qing

Abstract: Objective To discuss the antitumor mechanism preliminarily by observing effects of Celastrus orbiculatus extracts (COE) on vascular endothelial growth factor (VEGF) mRNA and protein expression in hepatoma (Hepa1-6) cells of mice. Methods Hepa1-6 cells were treated with COE at different nontoxic concentration (0, 10, 20, 40, 80, and 160 μg/mL) for 16 h. The mRNA and protein expressions of VEGF were detected by reverse transcription-PCR and Western Blotting, respectively. Results COE significantly inhibited VEGF expression at both mRNA and protein levels in a dose-dependent manner. Conclusion COE can inhibit VEGF expression in Hepa1-6 cells, therefore suggest that VEGF could be chosen as an therapeutic target for COE in the context of cancer chemoprevention and anticancer therapy.

Visualization of Multivariate Metabolomic Data

Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data, the quantitative concepts in both scores plot and loading plot are rather obscure. This article introduced three profiles that assisted the better understanding of metabolomic data. Methods The discriminatory profile, heat map, and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis. Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups, while the statistic profile showed the potential markers of statistic significance. Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.

Inhibition of Corydalis decumbens Alkaloids on Hydrogen Peroxide- induced Apoptosis of PC12 Cells through Down-regulating Caspase-3 Expression

YAN Ren-jie,YANG Yi-fang,LUO Yong-ming and WU Chun-zhen

Objective To extract alkaloids from Corydalis decumbens (AsCD) by supercritical CO2 fluid extraction (SFE) and to evaluate protective effects of AsCD against hydrogen peroxide (H2O2)-induced apoptosis in rat PC12 cells. Methods AsCD were extracted by SFE and oxidative damage PC12 cells model was induced by H2O2. The survival rate of the cells was determined by MTT assay; Lactate dehydrogenase release was determined by ultraviolet spectrophotometry; Flow cytometry was used to detect apoptosis; Caspase-3 mRNA and protein were determined by real-time PCR and Western blotting assay, respectively. Results AsCD remarkably reduced the cytotoxicity, prevented membrane damage, and inhibited cell apoptosis. AsCD inhibited Caspase-3 mRNA and protein expression induced by H2O2 in PC12 cells. Conclusion AsCD possess protective effects against H2O2-induced apoptosis in PC12 cells, and the mechanism of AsCD responsible to the inhibition of apoptosis is possibly attributed to the down-regulating Caspase-3 expression. AsCD might be useful in the treatment of oxidative stress-related neurodegenerative diseases.

“Dose-effect-response” Relationships of Paeoniae Radix Rubra on α-Naphthylisothiocyanate-induced Acute Cholestatic Hepatitis in Rats

Objective To investigate the hepatoprotective effects of Paeoniae Radix Rubra (PRR) at different doses against α-naphthylisothiocyanate (α-NIT)-induced acute cholestatic hepatitis in rats. Methods Rats were ig admini- strated with vehicle or PRR [(1, 9, 18, 36, 54, 72, and 81 g/(kg?d)] 3 d before and 2 d after α-NIT (60 mg/kg) ig administration. The general status of rats, histopathology of liver, serum alanine aminotransaminase, aspartate aminotransaminase, total bilirubin, direct bilirubin, and alkaline phosphatase levels, were observed at respective time points (24 and 48 h) after α-NIT administration. Using cluster analysis and correspondence analysis, the “dose-effect-response” relationships of PRR were evaluated. Results The results showed that compared with model group, the serum biochemistry index significantly decreased with the increasing of PRR dosage (P < 0.01), and the change and necrosis of hepatic cellula, and inflammatory cell infiltration were gradually alleviated. However, the improvement was not obviously found in the low-dose group [1 g/(kg?d)]. The cluster analysis and correspondence analysis results showed that different doses of PRR could significantly ameliorate α-NIT-induced acute cholestatic hepatitis of rats in a dose-dependent manner. Conclusion The experiments show that administration doses of PRR in clinical use should be added properly in order to gain the expectant therapeutic effect, especially in the treatment of heavy acute cholestasis hepatitis.

Simultaneous Determination of Silybin A and Silybin B in Rat Plasma and Pharmacokinetic Study

Objective To investigate the bioavailability and pharmacokinetics of silybin A and silybin B in rats, respectively. Methods Following iv and ig administration of silybin to 20 Wistar rats, the plasma samples were collected at different time points up to 12 h. Sample pretreatment was involved in one-step protein precipitation with acetonitrile. Silybin A and silybin B were simultaneously determined by LC-MS/MS. Results After ig dosing silybin 28, 56, and 112 mg/kg to rats, the t1/2β values were 5.48, 5.08, and 5.73 h for silybin A, and 4.56, 4.12, and 5.53 h for silybin B; The Cmax were 674.3, 1349.4, and 2042.5 ng/mL for silybin A, and 671.0, 1365.4, and 2066.2 ng/mL for silybin B; The Tmax were 0.20, 0.23, and 0.20 h for silybin A, and 0.20, 0.23, and 0.20 h for silybin B; The AUC were 454.4, 845.9, and 1219.5 h?ng/mL for silybin A, and 432.0, 817.1, and 1153.6 h?ng/mL for silybin B. The absolute bioavailabilities of silybin A and silybin B were 2.86% and 1.93%, respectively. Conclusion Silybin A and silybin B have very low bioavailability after ig administration, and there is no significant difference in the pharmacokinetic parameters between silybin A and silybin B, which indicates that the two diastereoisomers have similar pharmacokinetic behavior in rats.

Meta Analysis of Lentinan Injection plus Cisplatin in Treatment of Malignant Pleural Effusion

WANG Quan,HE Xi-ran,WANG Man-cai and TIAN Jin-hui

Objective To evaluate the efficacy and safety of lentinan injection plus cisplatin (LIC) in the treatment of malignant pleural effusion (MPE). Methods We searched the database of Cochrane Library, PubMed, EMBASE, ISI Web of Knowledge, Chinese Biomedical Literature Database, Chinese Scientific Journals Full-text Database, Chinese Journal Full-text, and Google Scholar, etc., up to February 28th, 2011 to identify randomized controlled trials (RCTs) about lentinan injection (LI) for MPE, evaluate the quality of the included studies, and analyze the data by Cochrane Collaboration’s RevMan5.0 software. Results Twenty-nine RCTs involving 1831 patients were included. Meta analysis results suggested that there were some differences when comparing LIC with control groups suffering from MPE, for LIC could improve the near-term curative effect and the quality of life to some extent. Besides, compared with chemotherapy alone, LI plus chemotherapy had an advantage in relieving adverse reactions, such as gastrointestinal reactions, myelosuppression, chest pain, and general malaise. Conclusion The current evidence indicates that LI may have adjuvant therapeutic effects for MPE.

Pharmacokinetic Study of Baicalein and Its Major Metabolites after iv Administration in Dogs

Objective To develop and validate a simple, rapid, sensitive, and reproducible HPLC method for simultaneous determination of baicalein and its metabolite baicalin in dog plasma and for the subsequent pharmacokinetic study after iv administration to dogs. Methods An accurate and reproducible HPLC-UV method was developed and validated for simultaneous determination of baicalein and baicalin in dog plasma, using luteolin as internal standard. The analytes were separated by an Agilent Zorbax SB-C18 column (250 mm × 4.6 mm, 5 μm) and the column temperature was maintained at 40 ℃. The mobile phase was a binary mixture of acetonitrile and water (27:73), containing 0.05% phosphoric acid in water, with a flow rate of 1.0 mL/min. The UV detector was set at 276 nm. Results Linear relationships were validated over the range of 0.05－25 μg/mL for baicalein and 0.05－20 μg/mL for baicalin. The intra- and inter-day precision values for all samples were within 8.0%, using relative standard deviation. This method was successfully applied to the pharmacokinetic studies in dogs after iv administration of baicalein. Baicalein was converted to baicalin quickly. Cmax values were 21.13 μg/mL at 0.05 h for baicalein and 1.57 μg/mL at 0.5 h for baicalin, areas under the plasma concentration-time curve were 4.97 h?μg/mL for baicalein and 0.63 h?μg/mL for baicalin, and the elimination half-life is 0.50 h for baicalein and 0.75 h for baicalin, respectively. Conclusion The method is able and sufficient to be used in drug metabolism and pharmacokinetic studies of baicalein.

Protection of Effective Component Group from Xiaoshuan Tongluo on Brain Injury after Chronic Hypoperfusion in Rats

Objective To investigate the protective effects of purified effective component group in extract from Xiaoshuan Tongluo (CGXT) formula on chronic brain ischemia in rats. Methods CGXT 75, 150, and 300 mg/kg or vehicle were ig administered daily for four weeks to rats with bilateral common carotid arteries ligation (BCCAL). From the day 24 to 28 after BCCAL, Morris water maze was performed to assess the learning and memory impairment of rats. Four weeks after BCCAL, brain gray and white matter damage were assessed. Results In Morris test, the mean escape latency of rats in the CGXT (150 and 300 mg/kg) groups was significantly shorter than that in the vehicle group. CGXT also attenuated the neuronal damage in hippocampus and cortex and reduced the pathological damage in the optic tract and corpus callosum. Conclusion CGXT could improve learning and memory impairment resulted from BCCAL in rats. These results provide the experimental basis for the clinical use of CGXT in stroke treatment and may help in investigation of multimodal therapy strategies in ischemic cerebrovascular diseases including stroke.

Comparative Study of Anti-oxidative Effects of Tibetan Folk Medicine Erigeron multiradiatus during Plant Growth

ZHANG Zhi-feng,LIU Yuan,LUO Pei and ZHANG Hao

Objective To explore the effects of a potential anti-oxidative plant, Erigeron multiradiatus (Asteraceae), plant materials from naturally distributed high-altitude populations at different stages of life cycle were collected. Methods Fifteen extracts obtained from the Ganzi region (Sichuan, China) were studied to assess their radical- scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals and reducing power ability. Moreover, considering that anti-oxidants and free radical scavengers can also exert protective effect on endothelial cells from oxidative injury, these extracts were also evaluated for their anti-oxidative activity against cellular injury in the cultured human endothelial cell line (ECV304) induced by hydrogen peroxide (H2O2). Results All the extracts had radical-scavenging and/or reducing power ability, and the most active extract was found during flowering whereas the lowest appeared during vegetative growth period. The accumulation of anti-oxidative compounds was found to be affected by the altitude of growth environment. Total flavonoid content assay was also performed to support this outcome. Furthermore, these extracts also exhibited different effects on attenuating H2O2-induced cytotoxicity and inhibiting lipid peroxidation and LDH leakage from endothelial cells. Conclusion E. multiradiatus may be an important natural anti-oxidant and this property may contribute to verifying the utilization of this plant in Tibet folk medicine.

Factors Affecting Embryogenic Callus Production and Plant Regeneration in Anther Culture of Bupleurum chinense

Objective To evaluate the influences of the genotypes, anther developmental stages, and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense. Methods The different effects such as four genotypes, plant growth regulators, and temperature condition were compared in the experiments. The histological study was performed with the process of the anther culture. Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao (ZCYH), Z4, and Z5 at the early- to middle-uninucleate stages, except for genotype ZPM1 at the tetrad stage. Cold pretreatment increased the production of the embryogenic callus, in which 4-day cold pretreatment improved the production of embryogenic callus from 0% to 2.2% and 5.0% for genotypes ZPM1 and ZCYH, respectively. No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The highest regeneration rate (34.6%) was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine (BA). The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis. Chromosome counting of regenerated plantlets showed mostly diploid plant (2n = 12) with only one haploid plant (n = 6). Because of the low rate of microspore embryo formation, we only tracked the process of embryogenesis from the connective tissue, instead of microspore by histological observations. Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system. This is the first report on the haploid plantlet through the anther culture of B. chinense.

Development of HPLC Method to Evaluate Drug-processing Technique of Eucommiae Cortex

WANG Jia-long,LIU Er-wei,WU Shuai,YIN Zhao-ye and ZHANG Yi

Objective To establish a RP-HPLC method investigate the processing technique and mechanism of Eucommiae Cortex. Methods The RP-HPLC method was applied to simultaneously determining six ingredients, geniposidic acid, geniposide, genipin, chlorogenic acid, (+)-pinoresinol-di-β-D-glucopyranoside, and (+)-syringaresinol-di-β- D-glucopyranoside, in the different processed barks of Eucommia ulmoides. Results The valid method with good accuracy could be well used to study the processing technique of E. ulmoides; Besides, target ingredients in E. ulmoide were decreased within 6 h when they were processed. Conclusion Established RP-HPLC is a reliable method which could be used to research the processing technique of the barks of E. ulmoides. Moreover, the result of this study could be provided with significant evidence of processed barks of E. ulmoides.

Improvement of Growth and Periplocin Yield of Periploca sepium Adventitious Root Cultures by Altering Nitrogen Source Supply

ZHANG Jian,GAO Wen-yuan,WANG Juan,LI Xing-lin and XIAO Pei-gen

Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source. Methods Firstly, the effects of nitrogen source (NH4+-NO3?) at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P. sepium were investigated, and growth and production media for the two-stage culture based on the above results were established. Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH4+-NO3? (1:2) and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source. By adopting a fed-batch cultivation strategy, the dry weight adventitious root, periplocin content and yield were increased by 136%, 108%, and 389%, respectively when compared with those of the control, reaching up to 8.13 g/L, 157.15 μg/g, and 1277.63 μg/L, respectively. Furthermore, it was found that in the process of two-stage culture, the adventitious roots grew thicker significantly after they were transferred into production medium directly. Conclusion The ultimate yield of periplocin in P. sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.

A New Natural Compound with Cytotoxic Activity from Tripterygium hypoglaucum

LI Wei,WANG Fang and ZHAO Yu-qing

Objective To find antitumor constituents from Tripterygium hypoglaucum. Methods The chloroform extract of T. hypoglaucum was separated by silica gel column chromatography and preparative HPLC. The structures of compounds isolated were identified by spectral analysis and chemical evidence. Results Seven compounds were isolated and identified as rhein ethyl ester (1), chrysophenol (2), physcion (3), emodin (4), wilfordine (5), wilforgine (6), and wilforine (7). The cytotoxic activities of the compounds against cancer cell lines were assayed. Conclusion Compound 1 is a new natural compound with strong activities against human cancer cell lines (A2780 and OVCAR-3). Compounds 2－4 are isolated from this genus plants for the first time. The possible structure-activity relationship among compounds 1－4 shows that the methoxy group or oxyethyl moiety might be responsible for the cytotoxity.

Smashing Tissue Extraction and HPLC Determination of Paclitaxel and 10-Deacetylbaccatin from Taxus x media

LIU Yan-ze,GAO Wen-qin,WANG Ji-wen,ZHANG Yu and ZHAO Yu-qing

Objective To optimize the extraction technology of Taxus x media by using the contents of Paclitaxel and 10-deacetylbaccatin (10-DAB), two representative active diterpene alkaloids of taxane type from T. x media, as evaluation standard. Methods The smashing tissue extraction (STE) of Paclitaxel and 10-DAB from T. x media, was investigated by comparing with ultrasonic extraction (UE) which was one of the modern technologies of extraction. Results STE was more efficient than UE, and the contents of 10-DAB and Paclitaxel in the extracts obtained by STE were higher than those by UE. Conclusion STE is a fast, high-performance, and energy-saving technology for the extraction of diterpene alkaloids of taxane type. STE also provides a simple, component-safe, workable, and highly efficient method for the extraction of active natural product.

Protective Effect of Wenxin Granula on Heart from Myocardial Infarction through Regulating Intracellular Ca2+

Objective To assess the anti-arrhythmic activity and cardioprotective effects of Wenxin Granula, a traditional Chinese formula (consisting of Salviae Miltiorrhizae Radix, Polygonati Rhizoma, Notoginseng Radix et Rhizoma, Nardostachyos Radix et Rhizoma, Angelicae Sinensis Radix, and Succinum), on heart in ischemic-induced myocardial infarction (MI) rats and compare with those of Amiodarone which have been demonstrated in clinic. Methods Rats were randomly divided into Sham-operated (control), MI + Amiodarone [5 mg/(kg?d)] (MI), and MI + Wenxin Granula [10 mg/(kg?d)] groups and left anterior descending coronary artery was occluded in each group. After left anterior descending for 12 h, standard lead II of administration electrocardiogram was recorded in order to analyze the occurrence of arrhythmia. After one month, the size of the infarct area of heart was evaluated by TTC staining method and haemodynamic function was assessed to detect the heart function. Laser scanning confocal microscope and the technique of patch clamp were used to detect the intracellular Ca2+ ([Ca2+]i) and L-type calcium current (ICa-L), respectively. Results Both Wenxin Granula [10 mg/(kg?d)] and Amiodarone [5 mg/(kg?d)] could markedly decrease the incidence of arrhythmia in heart of rats which were subjected to ischemic injury. After one month, Wenxin Granula could significantly decrease mortality to 22.22% and reduce the infarct area (P < 0.05), but Amiodarone did not. The mechanism may involve that Wenxin Granula attenuated [Ca2+]i decreasing in MI rats. Additionally, Wenxin Granula could obviously ameliorate the impaired heart function of MI rats by decreasing the elevated left ventricular end-diastolic pressure and increasing the attenuated maximum change velocity of left ventricular pressure in the isovolumic contraction or relaxation period. On the other hand, electrophysiological experiment results revealed that Wenxin Granula administration one month later also increased the reduced ICa-L density in rat ventricular myocytes in MI rats. The results of LSCM showed that Wenxin Granula could recover the amplitude of [Ca2+]i decreased by heart failure during long term. Conclusion Wenxin Granula could not only inhibit the incidence of arrhythmia but also decrease the mortality, which was accompanied by recovering the amplitude of [Ca2+]i. This protective effect of Wenxin Granula may partially be mediated through changing ICa-L as well as increasing [Ca2+]i.

Protein Tyrosine Phosphatase 1B Inhibitors from Plantago asiatica

CUI Long,LEE Hyun-sun,AHN Jong-seog,YUAN Guang-xin and SUN Ya-nan

Objective To identify the active compounds for protein tyrosine phosphatase 1B (PTP1B) from the seeds of Plantago asiatica. Methods Bioassay-guided fractionation resulted in the isolation of iridoid glucosides (1－5) with PTP1B inhibitory activity. Results Five compounds were identified as desacetylhookerioside (1), melittoside (2), geniposidic acid (3), 10-O-acetyl-geniposidic acid (4), and alpinoside (5). Conclusion Isolated compounds 3－5 inhibit PTP1B with IC50 values ranged from (16.3 ? 1.1) to (19.8 ? 1.2) μmol/L.

Feature Extraction of Chinese Materia Medica Fingerprint Based on Star Plot Representation of Multivariate Data

CUI Jian-xin,HONG Wen-xue,ZHOU Rong-juan and GAO Hai-bo

Objective To study a novel feature extraction method of Chinese materia medica (CMM) fingerprint. Methods On the basis of the radar graphical presentation theory of multivariate, the radar map was used to figure the non-map parameters of the CMM fingerprint, then to extract the map features and to propose the feature fusion. Results Better performance was achieved when using this method to test data. Conclusion This shows that the feature extraction based on radar chart presentation can mine the valuable features that facilitate the identification of Chinese medicine.

Study on Plasma Concentration and Bioavailability of Wogonin in Beagle’s Dogs

LI Jian-chun,CHEN Fei-hu,DONG Hai-jun and GAO Shu — 2011/7/1 0:00:00

Objective To develop an LC-MS/MS method for determining the concentration of wogonin in dog plasma and investigate the pharmacokinetics and bioavailability by different administrations of wogonin in Beagle’s dogs. Methods LC-MS/MS was employed in determining the concentration of wogonin with the selected ion monitoring model after liquid-liquid extraction with ethyl acetate of dog plasma samples. The lower limit of quantification was 0.105 μg/L. Target ions were at m/z 285.0→270.0 for wogonin and 373.3→305.3 for finasteride. In a randomized, self-control, and cross-over study, six male Beagle’s dogs were treated with different administration methods in three test periods. Pharmacokinetic parameters were calculated with DAS software (Ver. 2.0). Results The calibration curve was linear in the range of 0.105－107.36 μg/L for wogonin in dog plasma samples. The main pharmacokinetic parameters of ig administration (native drug of 15 mg/kg and solution preparation of 5 mg/kg) and iv route were as follows: Cmax (2.5 ± 1.1), (7.9 ± 3.3), and (6838.7 ± 1322.1) μg/L, tmax (0.7 ± 0.3) and (0.3 ± 0.2) h for the both former, AUC0-t (7.1 ± 2.0), (21.0 ± 3.2), and (629.7 ± 111.8) μg?h/L. The absolute bioavailability of native and solution of wogonin were (0.59 ± 0.35)% and (3.65 ± 2.00)%, respectively. Conclusion The validated method is convenient, sensitive, and specific, and the improvement of wogonin solubility could remarkably increase the absolute bioavailability.

Analgesic and Anti-inflammatory Effects of Ginger Oil

Objective Ginger (Zingiber officinale) is widely used as a spice in cooking and as a medicinal herb in traditional herbal medicine. The present study was to investigate the analgesic and anti-inflammatory activities of ginger oil in experimental animal models. Methods The analgesic effect of the oils was evaluated by the “acetic acid” and “hot-plate” test models of pain in mice. The anti-inflammatory effect of the oil was investigated in rats, using rat paw edema induced by carrageenan, adjuvant arthritis, and vascular permeability induced by bradykinin, arachidonic acid, and histamine. Indomethacin (1 mg/kg), Aspirin (0.5 g/kg) and Dexamethasone (2.5 mg/kg) were used respectively as reference drugs for comparison. Results The ginger oil (0.25－1.0 g/kg) produced significant analgesic effect against chemically- and thermally-induced nociceptive pain stimuli in mice (P < 0.05, 0.01). And the ginger oil (0.25－1.0 g/kg) also significantly inhibited carrageenan-induced paw edema, adjuvant arthritis, and inflammatory mediators-induced vascular permeability in rats (P < 0.05, 0.001). Conclusion These findings confirm that the ginger oil can be used to treat pain and chronic inflammation such as rheumatic arthritis.

RP-HPLC Determination of 1,3-Dideoxygalactonojirimycin in Bombycis Faeces

QI Hui,ZHAO Hui,DING Zhen-ying and ZHU Yuan-yuan

Objective To establish a simple and rapid method for the determination of 1,3-dideoxygalactonojirimycin in Bombycis Faeces, a potent glucosidase inihibitor, by HPLC. Methods A RP-HPLC method with fluorescence detection has been developed. Results The HPLC method developed in this research has a good reliability including accuracy and precision. The detection limit was less than 72 ng. Conclusion This method is sufficiently sensitive for determining 1,3-dideoxygalactonojirimycin in Bombycis Faeces and other related products.

Inhibition of Sanggenon G Isolated from Morus alba on the Metastasis of Cancer Cell

CUI Long,LEE Hyun-sun,OH Won-keun and AHN Jong-seog

Objective An organic layer prepared from the cortex of Morus alba (Moraceae) was studied in order to identify the active compounds for heparinase. Methods Bioassay-guided fractionation resulted in the isolation of sanggenon G. Results The compound showed inhibitory activity with IC50 of 3.7 μmol/L on heparinase in vitro as well as 24 μmol/L in invasion assay using MDA-MB231 cells. Sanggenon G also had the moderate cytotoxicity at SW 620 (colon) and ACHN (kidney) cancer cell lines with IC50 of 10.96 and 13.44 μmol/L, respectively. Conclusion This is the first time that prenylated flavonoid sanggenon G is described as heparinase inhibitor. Besides, this flavonoid would be expected to be a metastasis inhibitor of cancer cells and also a valuable reagent to explore the mechanism of heparinase/heparanase-mediated metastasis.

Antibacterial Mechanisms of Berberine and Reasons for Little Resistance of Bacteria

JIN Jian-ling,HUA Guo-qiang,MENG Zhen and GAO Pei-ji

Objective To study the antibacterial mechanisms of berberine and try to understand the reasons why bacteria cells difficultly resisted to it. Methods Detecting the minimal inhibitory concentration (MIC) of bacterial cultures incubated under sub-MIC concentration of berberine, Huanglian, and Neomycin for more than 200 generations, in order to analyze the bacteria resistance. Detecting the binding kinetics of berberine to DNA, RNA, and proteins. Observing the changes in bacterial cell surface structure with scanning electron microscopy. Detecting the Ca2+ and K+ released from berberine-treated bacterial cells with atomic absorption spectrum. Detection the absorption of methyl-3H-thymine (3H-dT), 3H-uridine (3H-U), and 3H-tyrosine (3H-Tyr) into berberine-treated bacterial cells. Results MICs of bacterial cultures, growing more than 200 generations in MH medium with 1/2 MIC of berberine (BA200) or Huanglian (HA200), did not increase compared to the control, while remarkably increased in MH medium with 1/2 MIC of Neomycin (NA200). In addition, from the culture NA200 it was easy to isolate resistant mutant strains which could grow in MH medium with more than four times MIC Neomycin, but from the culture BA200 and HA200 it was difficult to isolate berberine or Huanglian mutant strains could grow in MH medium with more than four times MIC berberine or Huanglian. The binding kinetics of berberine to DNA, RNA, and proteins illustrated that berberine could easily and tightly bind to DNA and RNA, and hardly dis-bind from DNA- and RNA-berberine complexes. Berberine could easily bind to protein too, but also easily dis-bind from berberine-protein complex. The bacterial cells treated with berberine sharply decreased the absorption of 3H-dT, 3H-U, and 3H-Tyr, as the radioactive precursors of DNA, RNA, and protein biosynthesis. Berberine could damage bacterial cell surface structure, especially for Gram-negative bacteria. Ca2+ and K+ released from berberine-treated cells increased significantly compared to the control. Conclusion All of above results indicate that bacterial cells could not easily become resistant mutants to berberine. The mechanisms for the bactericidal effect of berberine include: inhibiting DNA duplication, RNA transcription, and protein biosynthesis; influencing or inhibiting enzyme activities; destructing the bacterial cell surface structure and resulting in Ca2+ and K+ released from cells. All of the berberine bactericidal mechanisms are the most essential physiological functions for a live cell, if influenced any one such function, the mutation would be lethal mutation, so that it is difficult to get berberine resistant cells. The results in this paper also prefigure that berberine and its related Chinese medicines would provide a feasible way to control antibiotic resistance problem.

Antitumor Activity of Dichloromethane Extract from Salvia plebeia and Induction of Apoptosis on K562 Cells

REN Jie,PAN Sha-sha,LU Xu-zhang,ZHOU Min and HU Kun

Objective To study the antitumor activity of extract from Salvia plebeia and investigate whether the extract induce apoptosis of K562 cells. Methods The aqueous, petroleum ether, dichloromethane (CH2Cl2), ethyl acetate, and butanol extracts were prepared from the aerial parts of S. plebeia. Taking fluorouracil as reference, the cytotoxic activities of these extracts on HeLa, A549, SGC-7901, HCT-116, K562, LoVo, DU-145, and HepG2 cells were evaluated. To clarify the apoptosis of K562 cells induced by CH2Cl2 extract, the methods of Hoechst 33258 staining, flow cytometry assay, and DNA ladder assay were investigated. Results The CH2Cl2 extract showed the most potent cytotoxic effect against K562 cells, with an IC50 < 15 μg/mL for 3 d treatment. The characteristic apoptotic symptoms such as DNA fragmentation and chromatin condensation were also observed in the K562 cells. Conclusion The CH2Cl2 extract from S. plebeia may inhibit the cancer cell proliferation by inducing cell apoptosis.

Induction of Angiogenesis and Neurogenesis by Serum from Rats Treated with Shunaoxin Dropping Pills

Objective Shunaoxin Dropping Pills (SDPs), a Chinese patent medicine, has been used widely in China for the treatment of headache, amnesia, and insomnia. The aim of the present study is to observe the effect of SDPs on inducing angiogenesis and neurogenesis in vitro. Methods The present testing system using the serum obtained from animals ig treated with SDPs and a co-culture system in vitro was used to investigate if SDPs promotes brain microvascular endothelial cells (BMECs) tube formation and neural differentiation of neural stem/progenitor cells (NSPCs), which plays important roles in angiogenesis and neurogenesis. Results The SDPs serum sampled from rats ig treated with SDPs for 3 d dose-dependently promoted the tube like structure formation of cultured BMECs, and enhanced the fraction of MAP-2 positive cells of NSPCs, which co-cultured with the BMECs and astrocyte. In addition, there was no significant change in the percentage of glial fibrillary acidic protein positive cells. Conclusion Our results show that SDPs serum can induce neural differentiation and BMECs tube formation in vitro.

Anti-diabetic Activity of Zhenqing Recipe and Ligustri Lucidi Fructus in Type 2 Diabetic Rats

Objective To investigate the influence of Zhenqing Recipe (ZQR) and Ligustri Lucidi Fructus (LLF) on diabetic rats and its possible mechanism. Methods The model of type 2 diabetic rats was established by feeding a high-sucrose-high-fat diet and injecting a low dose of Streptozotocin in Wistar rats. The model rats were randomly divided into three groups: diabetic model, ZQR-treated, and LLF-treated groups for 8-weeks treatment. The normal Wistar rats were as a normal control group. Results The level of fasting blood glucose in ZQR and LLF groups was decreased compared with model group (P < 0.01, 0.05, respectively). Both ZQR and LLF markedly reduced serum triglycerides (P < 0.01, 0.05, respectively), and increased the insulin sensitivity index (P < 0.05). Histopathology revealed that ZQR and LLF reduced pancreatic damage. Immunohistochemistry evaluation showed that the percentage of insulin positive cells in pancreatic island was higher than model group (P < 0.01, 0.05, respectively).The mRNA and protein expression of SREBP-1c in pancreas were significantly decreased in ZQR and FLL group (P < 0.01). Conclusion ZQR has therapeutic effect on type 2 diabetes, it ameliorates the histopathological changes of pancreas, protects β cells, improves insulin resistance, and attenuates the expression of SREBP-1c. This study also provides the anti-diabetic evidence of FLL even its effects are weaker than ZQR.

Protection of Astragaloside Derivate on Oxidative Stress and Hypertrophy in Cardiomyocytes

HAO Chun-hua,WANG Wei-ting,ZHAO Zhuan-you and TANG Li-da

Objective The astragaloside IV (ASI) has been proved to play an important role in protecting against cell death on cardiovascular diseases. This study aims to investigate the effect of the astragaloside derivate (ASId) on confronting oxidative stress and hypertrophy in myocardial cells. Methods Following exposure embryonic rat cardiac H9c2 cells to hydrogen peroxide (H2O2) and angiotensin II for developing oxidative stress and hypertrophy, ASId at final concentrations (0.1, 1, and 10 μmol/L) was added to study its role in protecting cardiomyocytes by biochemical detection and cell size measurement. In addition, the mitochondrial permeability transition pore (mPTP) opener atractyloside (20 μmol/L) and inhibitor cyclosporin A (CSA) (1 μmol/L) were employed to investigate the possible mechanisms for anti-oxidation. Results ASId at 1 and 10 μmol/L in cultures suppressed oxidative stress at different degrees, which induced the decrease in LDH activity and MDA content, and also the increase in SOD activity in comparable with the model group; The mPTP opener atractyloside and inhibitor CSA weakened and strengthened the role of ASId, respectively. ASId at 10 μmol/L inhibited cell hypertrophy, and the cell diameter, surface area, and protein content were all decreased in comparable of those cells in model group. Conclusion ASId is involved in the cytoprotective effects on oxidative stress through a pathway mediated by mPTP, and also has a protective effect against hypertrophy.

A New Alkaloid from Bombycis Feculae and Its α-Glucosidase Inhibitory Activity

ZHU Yuan-yuan,WANG Zhi-hong,QI Hui and BAI Gang

Objective To study the chemical constituents of Bombycis Feculae. Methods Chemical constituents were isolated by HPLC-ELSD. The structures of the isolated compounds were determined by spectral means. Results Two compounds were isolated and identified as 1-deoxynojirimycin (1) and (2R,3R,5R)-2-(hydroxymethyl) piperidine-3,5-diol, named as 1,3-dideoxygalatonojirimycin (2). Conclusion Compound 2 is a new alkaloid. The extract of Bombycis Feculae, compound 1 and compound 2 show inhibitory activities against α-glucosidase.

Simultaneous Determination of Multiple Bioactive Constituents in Total Alkaloid of Sophora alopecuroides by HPLC-DAD

ZHAO Wen-chang and SONG Li-jun

Objective To develop a qualitative and quantitative simultaneous determination of multiple bioactive constituents in total alkaloid in Sophora alopecuroides (TASA). Methods In the experiment, a new and simple HPLC-DAD method for the simultaneous determination of multiple constituents in TASA was developed. The separation was performed on a Kromasil C18 column (250 mm × 4.6 mm, 5.0 μm) eluted with 0.02 mol/L potassium dihydrogen phosphate (adjusted pH 4.3 using 1% glacial acetic acid) and acetonitrile (75 : 25) at a flow-rate of 0.7 mL/min. The detection wavelength was set at 210 nm. Results Five constituents (sophoridine, matrine, oxymatrine, aloperine, and lehmannine) were simultaneously analyzed in this study. Four of them were identified and determinated by the developed method. The calibration curves exhibited linear regressions (r2 > 0.9995). The injection precision, the intra-day precision, and the analysis repeatability were validated with the RSD values less than 5.0%. The mean recoveries of the four constituents were ranged from 98.62% to 100.20%, and the RSD values were all less than 3.37%. Conclusion This method is convenient, fast, accurate, and is applicable to analyze the multi-constituents in TASA.

Simultaneous Determination of Four Major Steroidal Saponins in Seven Species of Dioscorea L. by HPLC-ELSD

SHEN Zhi,ZHANG Wen-ting,ZHAO Wei-liang and HUANG Wei

Objective To control the quality of the species in Dioscorea L. better. Methods An HPLC-ELSD method was developed for the first time to simultaneously determine four bioactive ingredients: dioscin, gracillin, protoneodioscin, and protoneogracillin in 31 samples belonging to seven species of Dioscorea L. from different areas. The column was an Inertsil HILIC (250 mm × 4.6 mm, 5 μm). The separation was carried out with a gradient program. The mobile phase was acetonitrile-water at a flow rate of 0.8 mL/min. Results The standard curve was rectilinear in the range of 0.464－12.97 μg (r = 0.9969) for dioscin, 0.310－7.09 μg (r = 0.9953) for gracillin, 0.469－11.66 μg (r = 0.9970) for protoneodioscin, and 0.276－6.87 μg (r = 0.9992) for protoneogracillin. The recoveries of the markers were 98.1%, 100.1%, 97.2%, and 96.4%, respectively. The contents of the four components were quite different among the seven species of Dioscorea L. Conclusion The proposed HPLC-ELSD method is convenient, fast, accurate, and applicable for simultaneous analysis of multiple bioactive components of species in Dioscorea L. for quality control, which could facilitate discovering new natural resources of steroidal saponin.

Smashing Tissue Extraction and GC Analysis of Active Fatty Acids from Oil Cake of Perilla Seeds

SUN Yan-ling,LIU Yan-ze,XIAO Han,WEI Ying-feng and ZHAO Yu-qing

Objective To optimize the extraction technology of perilla seeds oil from the oil cake of perilla seeds (OCPS) by using the contents of active fatty acids as evaluation standard. Methods The fatty acids were extracted from OCPS, the residue of perilla seeds after cold-press, by smashing tissue extraction (STE), the new technology selected through comparing with classical leaching extraction (LE), Soxhlet extraction (SE), ultrasonic extraction (UE), and supercritical-CO2 fluid extraction (SFE). For optimized condition of STE, orthogonal test was designed and completed. The contents of five fatty acids in extracted oil and OCPS were determined by GC. Results The optimized extraction parameters were smashing for 1.5 min under extraction power of 150 W and 1:6 of the material/solvent ratio. The contents of five fatty acids in the oils extracted by five techniques from OCPS and determined by GC were as follows: α-linolenic acid (41.12%－51.81%), linoleic acid (15.38%－16.43%), oleic acid (18.93%－27.28%), stearic acid (2.56%－4.01%), and palmitic acid (7.38%－10.77%). Conclusion The results show that STE is the most efficient technology with the highest yield (LE: 0.57%; SE: 1.03%; UE: 0.61%; SFE: 0.80%; STE: 1.17%) and shortest time (LE: 720 min; SE: 360 min; UE: 30 min; SFE: 120 min; STE: 1.5 min) among five tested extraction technologies. It is first reported using STE to extract herbal oil enriched with active fatty acids.

Quantitative Metabolomics: Analysis on Active Components in Extracts from Kaki Folium

DAI Li-peng,GU Yuan,YIN Ren-jie,LIU Chang-xiao and SI Duan-yun

Objective In order to analyze the active components in the extracts from Kaki Folium (KF), quantitative metabolomics approach was adopted to investigate the number of active components existing among the different extracts and their variation. Methods LC-MS method was established for the quantitative determination of the active components taking the mixture with reference substance as tested sample. Results In terms of the number of active components and amount presented in the different tested samples of KF extracted by many types of solvents, variation was observed. But rutin, astragalin, and kaempferol were presented in all samples. Difference was found between the samples extracted from the products on market and from the raw materials of KF processed by polar solvents with different recipes, however, the three active components were found in all samples examined. Conclusion These results might be valuable as all information could be used for the optimization of raw materials extraction procedure to enhance the productivity.

Material Base of in vivo Invigorating Qi and Enriching Blood of Bazhen Decoction by HPLC-ESI-MS

LI Wen-lan,BAI Jing,SUN Zhi,DU Juan and JI Yu-bin

Objective To identify the in vivo metabolized chemical constituents in Bazhen Decoction (BZD) and to study the relationship between Siwu Decoction (SWD) and Sijunzi Decoction (SJZD) with BZD. Methods Analysis and comparison were carried out by HPLC-ESI-MS. Serum samples after ig administration of preparations such as SWD, SJZD, and BZD, with different ingredients were collected for analysis. Results Twenty-two components were detected after ig administration of BZD, and six of them were metabolites and others were original form of the components contained in BZD. The prototype constituents were atractylenolide I, 5-hydroxymethyl-furoic acid (5-HMFA), oxypaeoniflorin, atractylenolide III, albiflorin, paeoniflorin, liquiritin, ferulic acid, ligustilide, and ginsenosides Rg1 and Rb1. The metabolized constituents were paeonimetabolin I, glycyrrhetic acid mono- glucuronide, glycyrrhetinic acid, and ginsenosides Rh1 and Rd. Conclusion The results also show that 5-HMFA, oxypaeoniflorin, albiflorin, paeoniflorin, ferulic acid, and ligustilide are common components absorbed into blood existing in both BZD and SWD, which are material base of enriching blood; Atractylenolide I, atractylenolide III, and liquiritin are common ingredients absorbed into blood existing in both BZD and SJZD, and they are material base of invigorating Qi. The results provide basic data for the further studies on the effective components, the effecting mechanism, and the quality control of BZD.

Protection of Total Flavonoid Fraction from Nervilia fordii on Lipopolysaccharide-induced Acute Lung Injury in Rats

Objective To investigate the effects of total flavonoid fraction (TFF) from Nervilia fordii on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats, and to explore their protective mechanism. Methods LPS-induced ALI model was established by LPS (5 mg/kg) injection via left cervical vein. Blood samples were collected from the cervical artery of all rats at 5 and 6 h after LPS challenge for arterial blood gas test and cytokines measurements, and pulmonary microvascular permeability (PMP), lung wet/dry weight ratio (W/D), and pathological features were observed. Results Phytochemical study showed that the TFF contained 67.3% of flavonoids expressed in rutin and three flavone glycosides. The TFF pretreatment (6.24 and 12.48 mg/kg) attenuated the partial arterial pressure of oxygen decline in blood significantly, and decreased the PMP and lung W/D ratio in ALI rats. In addition, the TFF (6.24 and 12.48 mg/kg) also ameliorated the LPS-induced lung damages including alveolar edema, neutrophils in?ltration, alveolar hemorrhage, and thickening of the alveolar wall. Furthermore, the treatment with the TFF (6.24 and 12.48 mg/kg) also down-regulated the levels of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and intercellular adhesion molecule-1 (ICAM-1), and up-regulated the level of anti-inflammatory cytokine IL-10 in serum of ALI rats simultaneously. Conclusion These results suggest that the TFF could protect LPS-induced ALI in rats, which may be mediated, at least in part, by adjusting the production of inflammatory cytokines including TNF-α, IL-6, ICAM-1, and IL-10.

Study on Hot Property of Aconiti Lateralis Radix Praeparata and its Compatibility with Zingiberis Rhizoma Based on Animal Temperature Tropism

Objective To establish an objective method for evaluating the intrinsic characteristics between hot and cold properties of herbal drugs and to study the hot property of Aconiti Lateralis Radix Praeparata (ALRP) and its compatibility with Zingiberis Rhizoma (ZR) on animal temperature tropism. Methods The equipment with cold/hot pads was used to investigate the variety of temperature tropism among mice treated by ALRP and its compatibility with ZR. Meanwhile, the activities of adenosine triphosphatase (ATPase), total anti-oxidant capability (T-AOC), and total superoxide dismutase (T-SOD) were measured. Results Compared with the homologous ALRP group, the compatibility between ALRP and ZR (AZ) had a stronger action against the decreasing of remaining rate on hot pad (P < 0.05), suggesting an enhancement of cold tropism. Meanwhile, the internal indexes of Na+-K+-ATPase activity, Ca2+-Mg2+-ATPase activity, and oxygen consuming volume increased significantly (P < 0.05), suggesting an enhancement of energy metabolism. The changes of T-AOC and T-SOD activity suggested that AZ reinforced the anti-oxidative capability of mice. In addition, it could be inferred from cluster analysis that the activity of AZ was concretely stronger than that of homologous ALRP. Conclusion ALRP and ZR are both herbs with hot property, but AZ has an enhanced trend of hot property. This external behavior of cold tropism and the internal activity of energy metabolism and anti-oxidant might reflect the internal hot property in an intuitive and objective way. The changes of ATPase activity of liver tissue might be the mechanism of drug action.

Ethnopharmacological Survey of Medicinal Plants in Baotou, Inner Mongolia, China

LI Min-hui,LIU Yue,WANG Zhen-wang,CUI Zhan-hu,HUANG Lu-qi and XIAO Pei-gen

Objective To document the knowledge and the use of indigenous medicinal plants by traditional healers in Baotou, Inner Mongolia, China. Methods Data were collected from 112 randomly-selected interviewees using semi- structured interviews in wild herbal plant collected from 2007 to 2010. The data from the interviewees were analyzed with two quantitative tools. With the informant consensus factor, the information homology was evaluated and with the fidelity level the most important species from the categories were found. Results One hundred and fifty-two species belonging to 112 genera in 48 families with medicinal values were recorded. The reported medicinal plants species were used to treat 63 kinds of diseases. And the medicinal plants in this district possessed significant potentials for their pharmacological activities in the context of ethnopharmacological knowledge, especially in the treatments of gastrointestinal, dermatological, and cardiovascular diseases. Conclusion In this work, 152 medicinal plants with their ethnopharmacological information are reported. This study demonstrates that many species play an important role in healing practices among inhabitants from Baotou. More ethnopharmacological information of Mongolian medicinal plants should be gathered and documented in further studies, which is a fundamental step toward developing efficacious natural drugs for various diseases.

Effect of Flavonoids in Morus indica on Blood Lipids and Glucose in Hyperlipidemia-diabetic Rats

MA Dong-qing,JIANG Zheng-ju,XU Shi-qiang,YU Xing,HU Xia-min and PAN Hong-yan

Objective To investigate the effect and possible mechanism of flavonoids extracted from Morus indica (FMI) on blood lipids and glucose. Methods The experimental hyperlipidemia-diabetic rats were induced by high-fat diet (HFD) and low dose of Streptozotocin (STZ). Flavonoids-treated rats were pretreated with FMI (50, 100, and 200 mg/kg). The plasma, skeletal muscle, and livers were isolated for biochemical assays, HE staining, immuno- histochemistry, and Western blotting analysis. Results The results showed that the body weight in flavonoids- treated (100 and 200 mg/kg) rats was reduced (P < 0.05, 0.01) compared to HFD-fed rats. FMI obviously reduced total cholesterol (P < 0.01), triglycerides (P < 0.05), and low-density lipoprotein cholesterol (P < 0.05), increased high-density lipoprotein (P < 0.05), and significantly decreased the atherosclerosis index (P < 0.01); FMI (100 and 200 mg/kg) also down-regulated the elevation of blood glucose induced by STZ (P < 0.05, 0.01); At the meantime, FMI increased hepatic superoxide dismutase (SOD) activity and reduced hepatic malondialdehyde (MDA) content obviously. In addition, the results showed that the expression of hepatic CYP2E1 was markedly decreased while the expression of GLUT-4 in skeletal muscles was increased by FMI. Conclusion The above results demonstrate that the effect of FMI is related to its up-regulation of hepatic SOD activity, reduction of hepatic MDA content, down-regulation of hepatic CYP2E1 expression, and increase of GLUT-4 expression in skeletal muscle, which suggests that FMI may prevent or improve hyperlipidemia and hyperglycemia caused by an excessive HFD.

Prophylaxis and Therapy of Isaria felina on Acute Renal Failure Induced by Glycerin in Rats

YANG Xi-hua,ZHANG Sheng-wan,REN Lian-sheng and BAI Xi-hua

Objective To evaluate the prophylaxis and therapy of Isaria felina (IF) on glycerin-induced acute renal failure (ARF) in rats. Methods Forty male Wistar rats were divided into control, model, Uremic Clearance Granule (UCG, positive control), high- and low-dose IF groups. Rats in the high- and low-dose IF groups were ig administered with 200 and 100 mg/kg IF, respectively, while rats were ig administered with 3.6 g/kg UCG successively for 7 d to establish UCG group. The rats in model, control, and drug-treated groups were im injected with 8 mL/kg 50% glycerin after drinking was quitted for 24 h to induce ARF in rats. The drugs were continued to give thereafter. The level of blood urea nitrogen (BUN) and serum creatinine (SCr) were determined 24 and 72 h after the injection of glycerin, also the kidney was dissected for pathology examination. Results Im injection with 8 mL/kg 50% glycerin could successfully induce ARF in rats. The dose of 200 mg/kg IF could reduce the high level of BUN and SCr, and ameliorate the pathological damage of the kidney. Conclusion IF has good protective and therapeutic effects on ARF and it is a potential and valuable Chinese herb for ARF.

Preparation of Glycyrrhetinic Acid Monoglucuronide by Selective Hydrolysis of Glycyrrhizic Acid via Biotransformation

Objective To search for the microorganisms which have the high selectivity of hydrolyzing glycyrrhizic acid (GL) into 18β-glycyrrhetinic acid-3-O-β-D-glucuronide (GAMG) without glycyrrhetinic acid (GA) byproduct. Methods GL was biotransformed by Aspergillus sp., the products were separated by chromatography on reverse phase C18 column and semi-preparative HPLC, and their structures were elucidated on the basis of HR-ESI-MS, 1D NMR (1H-NMR, 13C-NMR, and NOESY) and 2D NMR (1H-1H COSY, HSQC, and HMBC) spectral analyses. Results Aspergillus sp. could partially hydrolyze GL into GAMG (3), along with two minor byproducts, 3-O-β-D-glucurono- pyranosyl-18β-liquiritic acid (1) and 3-O-β-D-glucuronopyranosyl-24-hydroxy-18β-glycyrrhetinic acid (2). Conclusion Aspergillus sp. has the high selectivity of hydrolyzing GL into GAMG without GA byproduct and the yield of GAMG is about 60%. The complete assignments of 1H-NMR and 13C-NMR data for compounds 1 and 2 are reported for the first time.

Influence of Panax ginseng Continuous Cropping on Metabolic Function of Soil Microbial Communities

YING Yi-xin,DING Wan-long,ZHOU Ying-qun and LI Yong

Objective To investigate the influence of Panax ginseng continuous cropping on the carbon substrate metabolic activity of microbes in soils sampled from Dafang, Huangni, and Wulidi in Jilin Province, China. Methods Soil metabolisms of soil communities were characterized by community level physiological profiles using BIOLOGTM EcoPlate. Results Soils sampled from the three sites were analyzed and their metabolic activities were compared. Principal component analysis explored the significant variance in metabolic function of microbial communities in soils, though the Shannon index and the evenness index of them were similar. Futhermore, two principal components (PC1 and PC2), which contributed 67.83% and 10.78% of total variance, were extracted respectively. And also, substrates significantly correlated with PC1 and PC2 at the three sampling sites were identified. Conclusion Characteristic of soil is the primary factor influencing microbial communities, and P. ginseng continuous cropping has significant influence on microbial community. Though soil samples show similar microbial metabolic profiles, microbial communities in rhizosphere soil are changed obviously during the cultivation of P. ginseng, which would finally result in the unbalance of microbial community. Phytopathogens would gradually be the predominants in rhizosphere soil and make P. ginseng sick.

Study on Targeting and in vitro Anti-oxidation of Baicalin Solid Lipid Nanoparticles

Objective To prepare liver-targeted baicalin solid lipid nanoparticles (BSLNs) and to study their in vitro anti-oxidative activity. Methods BSLNs were prepared by emulsification ultrasonic dispersion method and characterized by transmission electron microscopy and laser particle size distribution; The tissue in vivo distribution was detected by pharmacokinetics; In vitro anti-superoxide dismutase (SOD) activity and reduction capacity of BSLNs were determined; The ability of removing hydroxyl radical was determined by phenanthroline-Fe2+ oxidation. Results The best prescription was baicalin-soybean lecithin-glyceryl monostearate-poloxamer 188 (1:5:15:30); The encapsulation efficiency and drug loading were 84.7% and 5.65%, respectively, mean size of particles was (68.6 ± 8) nm, Zeta potential was ?22.13 mV; The in vitro anti-oxidant results showed that BSLNs had a significant inhibitory effect on SOD and a strong reducing capacity as well as a removing hydroxide radical ability. The targeting rate of BSLNs was 6.931 for liver. Conclusion The results demonstrate that BSLNs could enhance the liver targeting ability and in vitro anti-oxidative activity significantly.

Smashing Tissue Extraction and HPLC Determination of Active Saponins from Different Parts of Panax notoginseng

SHI Sheng-ming,LIU Yan-ze,TAI Wen,CHEN Chang-qing and ZHAO Yu-qing

Objective To optimize the extraction technology used for extracting active saponins from the roots, fibrous roots, basal part of stems, root verrucae, fruits, flowers, stems, and leaves of Panax notoginseng based on the contents of ginsengsides Rg1, Rb1, and notoginsengside R1 as evaluation indexes. Methods Different parts of P. notoginseng were extracted by smashing tissue extraction (STE), ultrasound extraction, and reflux extraction. The contents of ginsengsides Rg1, Rb1, and notoginsengside R1 in 24 kinds of extracts were determined by HPLC-UV. Hypersil C18 column (200 mm × 4.6 mm, 5 μm) and acetonitrile-warter (20:80 for 30 min→45:55 for 18 min→70:30 for 2 min→80:20 for 10 min→100:0) were used; UV detector was set at 203 nm; The flow rate was set at 1.0 mL/min. Results STE was the most efficient technology with the highest yield of active saponins among the three tested extraction technologies. Conclusion STE is a fast, effective, and economical method to extract the active saponins from different parts of P. notoginseng. It could significantly shorten the extraction time and simplify the determination of the pre-processing work on identifying P. notoginseng. Such quick and effective extraction provides a powerful tool for analyzing P. notoginseng in the future.

Triterpene Glycosides from Sea Cucumber Holothuria scabra with Cytotoxic Activity

HAN Hua,LI Ling,YI Yang-hua,WANG Xiao-hua and PAN Min-xiang

Objective To study the new triterpene glycosides from sea cucumber Holothuria scabra with cytotoxic activity. Methods Triterpene glycosides from H. scabra were separated and purified by chromatography on DA-101, silica gel, and reversed-phase silica gel column, as well as RP-HPLC. Their structures were elucidated on the basis of spectral data and chemical evidence. Results Three triterpene glycosides were identified as scabraside D (1), fuscocineroside C (2), and 24-dehydroechinoside A (3). Their inhibition on P-388, A549, MKN-28, HCT116, and MCF-7 cells were significant. Conclusion Scabraside D (1) is a new triterpene glycoside, and compounds 2 and 3 are isolated from H. scabra for the first time. The glycosides 1－3 show the in vitro cytotoxicity against five human tumor cell lines in comparison to 10-hydroxycamptothecin.

Inhibition of Ptychopetalum olacoides on Acetylcholinesterase Isoforms in Brain of Mice

Objective To further characterize the acetylcholinesterase inhibitors (AChE-Is) pattern of Ptychopetalum olacoides ethanol extract (POEE) on the cytosolic globular monomer (G1) and membrane bound globular tetramer (G4) AChE isoforms in brain areas relevant for cognition. Methods The G1 and G4 AChE isoforms were prepared according to the reported methods and the determination of AChE activity used was adapted from colorimetric method. Results POEE mostly inhibited G1 in hippocampus (75%), and G4 in frontal cortex (58%) and striatum (75%) (P < 0.05). Kinetic analysis indicated that POEE-induced AChE inhibition in hippocampus was of a competitive nature for G1 but uncompetitive for G4. Conclusion Considering the high density of cholinergic projection to the cortex and striatum, and the usefulness of conserving cytosolic acetylcholine to replenish synaptic vesicles in a highly active cognition site such as hippocampus, we argue that this could be a desirable profile for a clinically relevant AChE-I.

Neuroprotection of n-Butanol Extract from Roots of Potentilla anserina on Hypoxic Injury in Primary Hippocampal Neurons

Objective To investigate the protective effect of n-butanol extract from the roots of Potentilla anserina (NP) on hypoxic hippocampal neurons in neonatal rats. Methods Primary cultured hippocampal neurons were pretreated with different concentration of NP (0.25, 0.0625, and 0.0156 mg/mL) before incubation in a low oxygen (0.1%) environment for 4 h. Cell viability was evaluated by Trypan blue staining assay. Lactate dehydrogenase (LDH) released by neurons into the medium was measured. The activity of superoxide dismutase (SOD) in cell cytosol was determined using nitroblue tetrazolium. Morphological changes and mitochondrial function were observed by transmission electron microscopy. Results Hypoxic injury could decrease the cells viability of neuron, enhance LDH release (P < 0.05), decrease SOD activity, and increase mitochondrial injury. Pretreatment with NP significantly increased cell viability, decreased LDH release (P < 0.05), promoted SOD activity (P < 0.05), and remarkably improved cellular ultra-microstructure compared with the model group. Conclusion NP could protect the primary hippocampal neurons from hypoxic injury by attenuating mitochondrial cell death.

Effects of Two Curcuminoids on Candida albicans

ZHANG Da,LUO Jiao-yang,YAN Dan,JIN Cheng,DONG Xiao-ping and XIAO Xiao-he

Objective To investigate and compare the action of curcuminoids on the causal pathogens of Candida albicans growth. Methods The effects of curcumin (CUR) and demethoxycurcumin (DMC) on C. albicans growth were first investigated and compared by microcalorimetry coupled with multiple analytical methods. The quantitative thermo-kinetic parameters obtained from these curves were analyzed to show difference of the actions. Results By analyzing the main parameters screened from principal component analysis together with 50% inhibiting concentration values, it was demonstrated that both CUR and DMC showed good antifungal activities and CUR was stronger. It was further concluded from structure-activity relationship that the existence of methoxy group might enhance lipophilicity of the mother nucleus, which made it easier for the molecular to enter into the cell membrane of fungi to inhibit its growth. Conclusion This study provides a new method for screening new antifungal agents with high efficacy and low toxicity. Meanwhile, it contributes to the application of curcuminoids as food additive, colorant, and drug. Microcalorimetry is real-time, online, and dynamic, and it could be used to characterize the subtle difference among the effects of synthetic and natural products on the vital process of fungi.

Pharmacokinetic Study on Hyperoside in Beagle’s Dogs

AI Guo,HUANG Zheng-ming and LIU Chang-xiao

Objective To develop and validate a simple, rapid, sensitive, and reproducible HPLC method for determination of hyperoside in plasma of dogs and for the subsequent pharmacokinetic (PK) study. Methods An accurate and reproducible HPLC-UV method was developed and validated for the determination of hyperoside in plasma of dogs, using kaempferol as internal standard. The plasma samples of dogs following ig administration of hyperoside were analyzed for the detection of quercetin after enzymatic hydrolysis treatment with combined β-glucuronidase and sulphatase. The analytes were separated on a Diamonsil C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-buffer solution (0.1 mol/L NH4Ac + 0.3 mmol/L EDTA-Na2)-acetic acid (60:40:1) and was delivered at a flow rate of 1 mL/min. The UV detector was set at 370 nm and the column temperature was maintained at 35 ℃. The sample injection volume was 20 μL. Data were collected and analyzed using the ANASTAR software. PK parameters were calculated with DAS software (2.0). Results Linear relationships were validated over the range of 0.01－1 μg/mL for hyperoside (r = 0.9997). The intra- and inter-day precision values for all samples were within 10.0%, and the accuracies of intra- and inter-day assays were within the range of 92.4%－102.4%. The validated method was successfully used to determine the hyperoside concentration in plasma of dogs for up to 12 h, after a single ig administration (25 mg/kg). The mean PK parameters for male and female dogs were as follows: Cmax (0.18 ± 0.05) and (0.16 ± 0.05) μg/mL, AUC0-∞ (0.79 ± 0.34) and (0.86 ± 0.27) μg/(mL?h), t1/2(ka) (0.89 ± 0.41) and (0.88 ± 0.28) h, respectively. Statistical analysis on the PK of hyperoside in male and female groups showed that sex had no significant impact on the PK of hyperoside (P > 0.05). Conclusion The method is able and sufficient to be used in drug PK studies of hyperoside.

Inhibition of Aloperine on Dextran Sulphate Sodium-induced Chronic Colitis in C57BL/6 Mice

SONG Li-jun,ZHAO Wen-chang and DENG Hong-zhu

Objective To investigate the effects of aloperine (ALO) on a model of dextran sulphate sodium (DSS)-induced chronic colitis in C57BL/6 mice. Methods Repeated colitis was induced by administration of four cycles of 4% DSS. The severity of colitis was assessed on the basis of clinical signs, ratio of colon weight and colon length, and histological grading scores. Moreover, secretory immunoglobulin A (S-IgA) and plasma haptoglobin (HP) were analyzed by enzyme-linked immunosorbent assay, and the changes of mRNA expression of ICAM-1 and MIF gene in colorectal tissue were detected by quantitative reverse transcriptase real-time polymerase chain reaction using SYBR Green I. Results ALO administration significantly attenuated the colon damage, caused substantial reductions of the rise in HP, and maintained the level of cecum S-IgA. ALO inhibited the ICAM-1 mRNA expression and had no effect on MIF mRNA expression. Conclusion The effect of ALO on DSS-induced chronic colitis in mice is investigated for the first time, which suggests that ALO could be an attractive therapeutic candidate in the treatment of inflammatory bowel disease.

Physicochemical Properties and Gastric Mucosa Irritation of Cantharidin-hydroxypropyl-β-cyclodextrin Inclusion Complex

AN Lin-na,DANG Yun-jie,HU Chun-hui and ZHU Chun-yan

Objective To increase the solubility and relieve the mucous irritation of cantharidin (CA) by preparing cantharidin-hydroxypropyl-β-cyclodextrin (CA/HP-β-CD) inclusion complex. Methods The inclusion complex was prepared by co-evaporation method and characterized by differential scanning calorimetry (DSC), X-ray diffractometry (XRD), and nuclear magnetic resonance (NMR). Results The disappearance of CA as well as the shift of exothermic peaks shown in DSC results indicated the complexation phenomenon. XRD results showed that the crystalline CA pattern had disappeared, and in NMR results, H-5 shifted from δ 3.731 to 3.695 after complexation and H-2 shifted from δ 3.626 to 3.598, which suggested that part of the drug had entered the HP-β-CD cavity to form an inclusion complex. The solubility increased 10.3 times after complexation and the mucous irritation of CA was relieved remarkably. Conclusion Through complexation with HP-β-CD, the solubility and dissolution rate of CA are improved significantly, and the irritation of musous is relieved.

Simultaneous Determination of Seven Alkaloids in Fufang Zhenzhu Tiaozhi Capsule by HPLC Coupled with DAD

CHEN Yuan-yuan,GUO Jiao,FAN Hui and HUANG Li-hua

Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids (magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine) in Fufang Zhenzhu Tiaozhi Capsule. Methods Chromatography was performed on a Dionex Acclaim C18 column (250 mm × 4.6 mm, 5.0 μm) at 30 ℃. The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution (0.015 mol/L, 40:60, including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0), with a flow rate of 1.0 mL/min. The detection wavelength was 270 nm. Results The calibration curves of magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine were linear in the range of 1.07－10.65, 0.78－7.55, 0.75－7.50, 1.60－15.95, 2.69－26.85, 2.31－23.10, and 6.04－60.40 mg/mL. The average recoveries of magnoflorine, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, and berberine were 101.0%, 101.2%, 100.1%, 100.0%, 100.1%, 101.1%, and 99.7%, respectively. Conclusion The method could be used for the quantitative determination of the preparation.

Standardization and Identification of Minor Components of Silymarin (MK-001)

LIU Yan-ze and LEE David Yue-wei

Objective To develop a highly effective HPLC method for standardization of silymarin and for characterization of minor components, which has not been reported previously. Methods An HPLC fingerprinting method and 1H-NMR spectroscopy were employed to provide assurance for a standardized silymarin product (MK-001). Results A total of 18 marker compounds were identified and characterized including ten flavonolignans and eight small molecules including adenine, adenosine, uridine, and 3,5,7-trihydroxychromone which were first isolated and characterized from silymarin. The HPLC fingerprinting method and 1H-NMR had been established for complete assignments of HPLC peaks with intensity over 1.0%. Conclusion The successfully established HPLC fingerprinting method and 1H-NMR could be applied for the standardization of commercial silymarin products. MK-001 represents the first standardized silymarin with the highest content of flavonolignans ( > 90%).

HPLC Fingerprint and LC-TOF-MS Analysis on Extract from Roots of Gentiana macrophylla

SU Qi,SHANG Ping-ping,ZHANG Yong-min,JIA Na,HE Jiao,ZHAO Wen-na and SUN Wen-ji

Objective Establishing a ?ngerprint method to identify the characteristic chemicals in the roots of Gentiana macrophylla and evaluate their quality. Methods RP-HPLC was developed for ?ngerprint analysis and determination of four ingredients in G. macrophylla roots from different sources. LC-ESI-TOF-MS was employed to identify the chromatographic peaks of the ?ngerprint. Results Five common peaks were identified by comparing their retention time with reference secoiridoid glucosides. Eight major peaks in chromatographic fingerprint were analyzed by on-line LC-ESI-TOF-MS. Four secoiridoid glucosides were identified based on their MS data. Conclusion The method is specific and could be served for the quality identi?cation and comprehensive evaluation of G. macrophylla.

Simultaneous Determination of Seven Flavonoids in Aerial Parts of Artemisia frigida by HPLC

WANG Qing-hu,AO Wu-li-ji and TAI Wen-quan

Objective To establish an HPLC method for the determination of seven flavonoids from the aerial part of Artemisia frigida. Methods Hypersil ODS-2 (300 mm × 4.6 mm, 5 μm) column was used, with acetonitril-0.2% phosphoric acid (gradient elution) as a mobile phase, and the detection wavelength was at 283 nm with flow rate at 1 mL/min. Results All calibration curves showed good linear regression (r > 0.9990) within the tested range. All average recovery was more than 98.00% and RSD was less than 3.0% (n = 6). Conclusion The method is steady and with good repeatability, and could be used to determine the content of flavonoids in A. frigida from different areas.

Optimization of Smashing Tissue Extraction Technology of Schisandra chinensis Fruits by Orthogonal Test

TANG Yun,LIU Yan-ze,HAN Ling and ZHAO Yu-qing

Objective To optimize the extract technology of active lignins from the fruits of Schisandra chinensis. Methods The content of schizandrin, gomisin A, and deoxyschizandrin were selected as standards to evaluate the efficiency of smashing tissue extraction (STE). Solid-liquid ratio, extracting times, ethanol concentration, and extracting time were investigated through orthogonal test. Results The optimized conditions for STE were ten times amount of 80% EtOH, extracting for three times, and 2 min for each time. Conclusion STE could obtain relatively higher yield, simplicity of operation, and benefit for environment protection. It could be better choice for the extraction of S. chinensis.

In vitro Metabolism of Strychnine by Human Cytochrome P450 and Its Interaction with Glycyrrhetic Acid

LIU Li,XIAO Juan,PENG Zhi-hong,WU Wen-hua,DU Peng and CHEN Yong

Objective To investigate the metabolism of strychnine (STN) and the metabolic interaction between STN and glycyrrhetic acid (GA) in vitro. Methods Human liver microsomes (HLM) and human recombinant cytochrome P450 (CYP) isoforms were employed to study the metabolism of STN and the metabolic interaction of STN with GA in vitro. Results In HLM, the Km, Vmax, and clearance of STN were 88.50 μmol/L, 0.88 nmol/(mg?min), and 9.93 mL/(mg?min), respectively. STN was metabolized mainly by CYP3A4. However, STN noncompetitively inhibited CYP3A4-catalyzed testosterone 6β-hydroxylation with IC50 value of 5.9 μmol/L and Ki value of 5.5 μmol/L. Moreover, GA competitively inhibited STN metabolism with IC50 value of 10.6 μmol/L and Ki value of 17.7 μmol/L. Conclusion Although STN is mainly metabolized by CYP3A4 in vitro, STN has noncompetitive inhibition on CYP3A4-catalyzed testosterone 6β-hydroxylation. Moreover, GA could competitively inhibit STN metabolism. The present work is helpful to elucidate the metabolic interaction between STN and GA.

Antihepatocarcinoma Effect of Solanine and Its Mechanisms

JI Yu-bin and GAO Shi-yong

Objective To explore the antitumor effect of solanine and its mechanisms. Methods The in vivo antitumor effect of solanine was observed using models developed through in vivo transplantation of tumor cells; In vitro lines of sensitive antitumor cells were selected from the digestive system using MTT assay; The effect of solanine on cell morphology was observed using transmission electronic microscopy; The morphology of apoptotic cells was observed using Annexin V/PI double staining and laser confocal scanning microscopy (LCSM); The rate of cell apoptosis was measured using Annexin V/PI double staining and flow cytometry; The concentration of intracellular Ca2+ ([Ca2+]i) was determined using Fluo-3/AM staining and LCSM; The membrane potential of cellular mitochondria was determined using TMRE staining and LCSM; The protein expression of Bcl-2 and Bax was measured using immunological marking and LCSM; And the activity of caspase-3 was measured using the colorimetric method. Results Solanine could inhibit the growth of tumor weight in S180 tumor-bearing mice and prolong the survival time of H22 tumor-bearing mice. MTT assay revealed that HepG2 cells were quite sensitive to solanine because solanine could induce morphological changes in HepG2 cells, with the rate of early apoptosis being 4%, 8.5%, and 20.1%, for HepG2 cells treated for 24 h with solanine at concentration of 0.4, 2, and 10 μg/mL, respectively. Solanine could raise the [Ca2+]i and lower the membrane potential. It could reduce the protein expression of Bcl-2 while increase that of Bax, thus increasing the activity of caspase-3. Conclusion The obvious antitumor activity of solanine in human hepatocarcinoma is demonstrated. This inhibitory effect is achieved through solanine decreasing the Bcl-2/Bax ratio, thus increasing [Ca2+]i, which could enhance the enzymatic activity of the caspase family, thus inducing the apoptosis of HepG2 cells.

Effects of Total Alkaloids in Buxus microphylla Leaves on Aorta Smooth Muscle of Rats and Their Mechanisms

ZHANG Hui-qin,LIU Yan-yan,LI Yong-wen,LI Li and CUI Zhi-qing

Objective To investigate the effects of total alkaloids in Buxus microphylla leaves (ABML) on isolated rats thoracic aorta rings, and then to explore the possible mechanisms underlying the effects. Methods Thoracic aortas of Wistar rats were isolated, removed, and mounted onto an organ bath. The effects of ABML at different concentration on the contraction of isolated thoracic aorta rings (with and without endothelium) precontracted with KCl or PE were observed with organ bath technique. Dose-effect curves of CaCl2 were recorded by organ bath technique. The concentration of intracellular Ca2+ ([Ca2+]i) increased by PE, KCI, and caffeine in the presence of ABML was determined using Ca2+ sensitive fluorescence indicator Fura-2/AM loaded thoracic aorta vascular smooth muscle (VSM) cells of rats. Results In aorta rings precontracted with PE and KCl, ABML produced concentration- dependent relaxation in both intact and denuded endothelium ring groups. There was no difference in the inhibition of contraction between the intact and denuded endothelium ring groups at the same concentration. Exposure of isolated thoracic aorta rings to ABML led to a significant reduction in the contracting response induced by CaCl2, and shifted the cumulative concentration-response curves to right. ABML could significantly inhibit the extracellular Ca2+ influx induced by PE and KCl under [Ca2+]0 of 1.5 mmol/L, with inhibitory ratios of 40.2% and 49.9%, respectively. In the case of Ca2+-free, ABML could significantly inhibit the intracellular Ca2+ release induced by PE, with inhibitory ratio of 72.4%. Conclusion ABML relaxes thoracic aorta VSM cells by suppressing influx of extracellular Ca2+ via voltage-dependent Ca2+ channel and receptor-operated Ca2+ channel.

Effect of n-butanol Extract from Potentilla anserina on Hypoxia- induced Calcium Overload and SERCA2 Expression of Rat Cardiomyocytes

Objective To investigate the effect of n-butanol extract from Potentilla anserina (NP) intervention on hypoxia-induced Ca2+ overload and SERCA2 expression of rat cardiomyocytes. Methods Primary cultured myocardial cell from SD neonatal rat (1－3 d) was used in the establishment of hypoxia model. After hypoxia for 3 h, the Ca2+ concentration of myocardial cells was measured with fura-2/AM fluorescent probe, and the biochemical indicator intracellular Ca2+-ATPase was examined and the mRNA and its protective protein levels of the sarcoplasmic reticulum (SR) Ca2+-ATPases (SERCA2) were assayed with RT-PCR, Western-blotting, and immune-cytochemical staining in each group. Results The results showed that NP decreased Ca2+ concentration, increased the activity of Ca2+-ATPase, and improved the mRNA and protein expression of SERCA2 in hypoxia-injured myocardial cells as compared with the model group. Conclusion These results indicate that NP could attenuate the Ca2+ overload. The mechanism might be explained as that NP could elevate the SERCA2 level, increase the activity of myocardium in rats, and further enhance the capacity of SR Ca2+ re-uptake.

Rational Daily Administration Times of Yinchenhao Decoction in Rats with Jaundice Based on PD/PK

Objective To study the rational daily administration times of Yinchenhao Decoction (YCHD) when it was used to treat experimental jaundice in rats based on pharmacodynamics/pharmacokinetics model. Methods Rats were modeled by 4% 1-naphthylisothiocyanate (75 mg/kg) for 48 h, then YCHD was drenched with doses of 0.324 g/kg (extract, calculated with the clinical dosage) once, 0.162 g/kg twice, and 0.108 g/kg thrice a day, respectively. The total bile and the flow rate of bile were observed after the first administration; Blood samples collected from the orbital sinus at different intervals were used to investigate the levels of liver enzymes (ALT and AST) and bilirubins (TBIL and DBIL), and determine the concentration of 6,7-dimethoxycoumarin (DME) in the plasma using UPLC at the same time, then we obtained the time-effect and time-dose curves. The rational daily administration times of YCHD when treating experimental jaundice were determined based on the comprehensive analysis of time-effect and time-concentration relationships. Results Within 10 h the total bile of rats which were administered once daily (G1) was 1.65 and 1.33 times higher than that of twice and thrice (G2 and G3) a day, respectively, and the four biochemical indexes (TBIL, ALT, DBIL, and AST) of G1 decreased faster than those of G2 and G3 (P < 0.05). On the other hand, the blood drug level of DME when administrated once daily could maintain at a higher level for a longer time, and its Cmax and AUC0→t were higher than those of G2 and G3, which might be the main reason why its effect was the most significant. Conclusion It is more appropriate to administrate once daily when YCHD is used to treat jaundice.

A Quantitative Method for Simultaneous Determination of Four Anthraquinones with One Marker in Rhei Radix et Rhizoma

ZHU Jing-jing,WANG Zhi-min,MA Xin-yu,FENG Wei-hong and ZHANG Qi-wei

Objective To develop a quantitative method for simultaneously determining multi-components in Rhei Radix et Rhizoma using one chemical reference substance. Methods The contents of multi-components were calculated by the UV relative correction factors (RCFs) of chrysophanol, physcion, and rhein to emodin. Results The values of RCFs at 274 nm for rhein, chrysophanol, and physcion to emodin were 0.712, 0.674, and 1.051. The calibration curves were linear over the ranges of 0.02－4.08, 0.02－4.12, 0.07－12.92, and 0.02－3.68 μg/mL for rhein, emodin, chrysophanol, and physcion, respectively. The contents of emodin in 18 samples were determined by the external standard method, and the contents of the other three anthraquinone aglycones were calculated according to their RCFs. Conclusion No significant difference is found in comparison with the classical method, indicating that the RCFs have high reliability within their linear ranges and could be used in quality control of Rhei Radix et Rhizoma. The quantitative analysis of multi-component with a single marker is especially suitable for herbal medicines containing unstable or hard to be purified components as quality control markers.

Simultaneous Determination of Six Quaternary Ammonium Alkaloids in Coptidis Rhizoma by UPLC

Objective To establish a new, rapid, and reliable reversed-phase ultra performance liquid chromatography (RP-UPLC) method for the simultaneous determination of six quaternary ammonium alkaloids (QAAs) in Coptidis Rhizoma. Methods The effect of different experimental parameters on the analysis of QAAs by RP-UPLC was evaluated. Results Optimal resolution was achieved with an Acquity UPLC BEH C18 column using a gradient elution profile and a mobile phase consisting of water spiked with 10 mmol/L ammonium bicarbonate (A, pH adjusted to 10.0 by ammonia water) and acetonitrile (B), at a flow rate of 0.30 mL/min and wavelength of 345 nm. The column temperature was set at 30 ℃. The proposed method was found to be reproducible, precise, and rapid according to the method validation. Conclusion The proposed method, which is compatible with MS analysis and the preparation of QAA, provides some helpful insights into the quality control of Coptidis Rhizoma.

Preparation, Pharmacokinetics, and Tissue Distribution Properties of Icariin-Loaded Stealth Solid Lipid Nanoparticles in Mice

LIU Ke-pan,WANG Li-feng,LI Yang,YANG Bin,DU Chao and WANG Yang

Objective To evaluate the difference of the pharmacokinetic (PK) and tissue distribution properties in mice administrated with lyophilized icariin stealth solid lipid nanoparticles (Ica-SSLN) modified by polyethylene glycol and icariin control solution (Ica-Sol). Meanwhile, to establish a sensitive, specific, and stable HPLC method for the determination of Ica in mice plasma and various tissues. Methods Ica-SSLN was prepared by high temperature melt-cool solidification method. Particle size and Zeta potentials were measured by a ZetaPlus. After iv administration of Ica-SSLN and Ica-Sol at a single dose of 7.46 mg/kg, the blood and tissues including brain, liver, spleen, lung, heart, and kidney were collected at different time points. The obtained concentration from HPLC analysis was statistically treated to determine the PK model and the relevant PK parameters using DAS1.0. Tissue distribution studies of Ica-SSLN were carried out in Kunming mice after iv administration and compared to Ica-Sol. Results The characteristic data showed that the mean particle size of Ica-SSLN was (50.03 ± 0.90) nm, entrapment efficiency was (71.67 ± 1.09)%, and the particles carried negative charge, Zeta potential was (?22.77 ± 1.89) mV. The concentration-time profiles of Ica in mice after iv administrated with Ica-SSLN and Ica-Sol were shown to fit a two-compartment open model. Compared with Ica-Sol, the t1/2β of Ica-SSLN was prolonged by seven times and the AUC was increased by four times. The levels of Ica concentration in the kidney tissues were significantly increased. In addition, compared with Ica-Sol, the relative target efficiency to kidney tissue was 79% and the relative tissue exposure was 16.95. Conclusion It demonstrates that Ica-SSLN has selective targeting to kidney tissue and the kidney targeted Ica-SSLN seems to have significant advantages and good development value.

Quantitative Proteomic Analysis of Bromotetrandrine and Tetrandrine in K562 Cell Line Using 18O-labeling Method

TAN Ying,GE Zhi-qiang and LIU Chang-xiao

Objective To compare quantitative proteomic analysis of bromotetrandrine (W198) which was a Class I new antitumor drug in China and tetrandrine (Tet) in K562 cell line using 18O-labeling method. Methods To illustrate its mechanism, a shotgun quantitative proteomic strategy employing 2D LC-MS-MS and trypsin catalyzed 18O-labeling quantification was carried out in this study. Compared to normal chronic leukemia cell line K562 and K562 induced by Tet, the proteomic changes of K562 induced by W198 were investigated. In order to validate the quantitation by the 18O-labeling, the analysis was done on an equivalent sample composed of the same amount of labeled and unlabeled proteins from normally cultured cells to act as a reference to the comparative sample. Results A threshold of ± 2-fold change for deciding whether a protein concentration was changed was settled for the following experiments. Comparing the 105 identified soluble proteins’ expression levels of the apoptosis starting up K562 cells after W198 induction with the normally cultured cells, 16 proteins were found with significantly altered expression levels after W198 treatment. Eight proteins were up-expressed including HMGB2, peroxiredoxin-2, and eIF4A-I, etc. Eight proteins were down-expressed including TCP-1, GRP94, GST-π, and SFGHs, etc. Compared to K562 induced by Tet, eight proteins of K562 were found with significantly altered expression levels after W198 treatment. Five proteins were up-expressed including HSP 90-β and 40S ribosomal protein S15a, etc. Three proteins were down-expressed including phosphoglycerate kinase 1, isoform 5 of interleukin enhancer-binding factor 3, etc. Conclusion The 18O-labeling MS-MS-based method is ideal as a discovery tool, but it is not suitable for validation using a large number of samples. Other more effective methods, such as Western blotting should be used for further validation of candidate cancer proteins discovered from 18O-labeling samples. In total, 105 soluble proteins were discovered, and 16 proteins were found with significantly altered expression levels after W198 treatment. These repressed or activated proteins are the potential drug targets of W198, which may provide novel targets for future development of biomarkers for cancer therapy.

Pharmacokinetic Effects of Baicalin on Cerebral Ischemia-reperfusion after iv Administration in Rats

MA Shu-wei,ZHAO Ming,LIU Hong-xia,WANG Li-yan and ZHANG Xian-tao

Objective To investigate the pharmacokinetic effects of baicalin on cerebral ischemia-reperfusion (I/R) after iv administration in rats. Methods The cerebral I/R rats were induced by occluding the bilateral carotid arteries of normal rats for 2 h, followed by reperfusion. The resultant animals were immediately iv administrated with baicalin (90 mg/kg), whilst the same dose of baicalin was injected to the normal rats. Plasma samples were collected at different time to construct pharmacokinetic profiles by plotting drug concentration vs time. Quantification of baicalin in rat plasma was achieved using a simple and rapid HPLC method. Results In normal rats, the major parameters of distribution half-life, elimination half-life, area under the plasma concentration-time (AUC), apparent volume of distribution (Vd), and clearance (CL), estimated by an open two-compartmental model, were 0.8868 min, 26.0968 min, 149.6204 mg/min?L, 4.765 L/kg, and 0.5776 L/ kg?min), respectively. However, in I/R rats, the corresponding parameters were 2.084 min, 34.4998 min, 260.0188 μg?min/L, 5.9376 L/kg, and 0.334 L/(kg?min), respectively. Conclusion The cerebral I/R could significantly increase AUC and Vd values, decrease CL values, and prolong the terminal half-life of baicalin. These findings suggest that the injuries of I/R could play an important role in pharmacokinetic process of baicalin.

Gastroprotective Effects of Ascaridole on Gastric Ulcer in Rats

Objective To evaluate the gastroprotective activity of ascaridole. Methods The gastroprotective effect of ascaridole was evaluated on ulcer healing in rats with acetic acid-induced chronic gastric ulcer, pylorus ligation- and Aspirin- induced gastric ulcer. Ascaridole was ig administered with the dosages of 10 and 20 mg/kg once daily for 7 d. Results Ascaridole showed the significant anti-ulcer effects. In acetic acid-induced gastric ulcer rats, the ulcer areas after 10 and 20 mg/kg of ascaridole treatment were (65.1 ± 20.0) and (50.6 ± 11.0) mm2, respectively, which were significant lower (P < 0.01) than that of the control group [(116.7 ± 35.8) mm2]. For pylorus ligation model, ascaridole showed a gastric ulcer healing effect in a dose-dependent manner. Ascaridole at the dose of 20 mg/kg showed 50% ulcer protection and had a significant (P < 0.05) gastroprotective activity since it decreased the total acidity and pepsin activity. Compared to the control group, the two dosages of ascaridole showed the significant reduction (P < 0.05) in the ulcer index on Aspirin-induced ulcer. Conclusion This study provides evidence that ascaridole shows potential efficacy on the healing of gastric ulcers induced by acetic acid, Aspirin, and pylorus ligation.

Analysis on Volatile Constituents in Leaves and Fruits of Ficus carica by GC-MS

LI Jun,TIAN Yu-zeng,SUN Bao-ya,YANG Dan,CHEN Ji-ping and MEN Qi-ming

Objective To identify and analyze the volatile constituents in the leaves and fruits of Ficus carica. Methods Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were used. Results The major components detected in volatile oil of the leaves were psoralen (10.12%), β-damascenone (10.17%), benzyl alcohol (4.56%), behenic acid (4.79%), and bergapten (1.99%), etc. The major components detected in volatile oil of the fruits were furfural (10.55%), 5-methyl-2-furaldehyde (10.1%), and benzeneacetaldehyde (6.59%), etc. Conclusion A total of 121 volatile constituents are identified in the leaves and 108 in the fruits of F. carica, among which 103 constituents are identified for the first time in the leaves and 100 in the fruits. Eighteen volatile constituents are identified in both leaves and fruits.

A New Homogenizing Technology to Obtain Rosmarinic Acid from Perilla Oil Meal

TANG Wei-zhuo,LIU Yan-ze and ZHAO Yu-qing

Objective To optimize the extraction technology of the active component, rosmarinic acid, an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid, in perilla oil meal for the first time by a new homogenizing technology called smashing tissue extraction (STE). Methods Orthogonal design was used to optimize the extraction condition. The content of rosmarinic acid was quantified from the methanol crude extract with the help of HPLC. Results The optimization of STE process to get rosmarinic acid from the perilla oil meal was the ratio of liquid to solid material at 10:1 and the power of extraction at 150 V, extracting twice (2 min for each time). Conclusion STE could be applied to extracting the active ingredients from the oil meals due to its high extraction efficiency. This new homogenizing technology has advantages on saving extraction time, raising extraction efficiency, and maintaining the temperature sensitive constituents.

Protection of Water Extract from Paeoniae Radix Rubra against Myocardial Ischemia in Mice Induced by Isoproterenol

WANG Zhi-bao,YANG Hui and GUO Chun-yan — 2013/11/15 10:48:51

Objective To study the protective effect of the water extract from Paeoniae Radix Rubra (WEPRR) against myocardial ischemia in mice induced by isoproterenol (ISO). Methods The mice were randomly divided into six groups: positive control, normal control, model, low-, mid-, and high-dose [0.3, 0.6, and 1.2 g/(kg·d)] WEPRR groups. The mice in WEPRR groups were ig administered with WEPRR, the mice in the positive control group were ig administered with 0.98% Di’ao Xinxue Kang [(0.195 g/(kg·d)], and the mice in normal and model groups were ig administered with the same volume of physiological saline once daily for consecutive 11 d. On the day 7 from the beginning of the ig administration, the mice in the model and WEPRR groups were ip perfused with 0.02 g/(kg·d) ISO. After 15 min of the last medication, the mice were anesthetized with isoflurane gas, the blood was collected through venous sinus of eye orbit, then the mice were killed. The heart tissues were rapidly removed from the mice, washed in physiological saline, soaked in filter paper, and stored in ?80 ℃ until use. The activities of lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB), creatine kinase (CK) in serum, superoxide dismutase (SOD), and the contents of malondialdehyde (MDA) in the heart of mice were determined, respectively. Results Compared with the model group, the activities of LDH, CK-MB, and CK in serum, and the SOD of mice in the positive control and WEPRR groups were increased and the content of MDA in heart was decreased. Conclusion The WEPRR has the cardioprotective activities on ISO-induced myocardial ischemia.

Effect of Cassia nomame on Small Intestine Movement, Diuresis, and Anti-inflammation in Rats

WANG Dan,WANG Wei-ning,QI Bao-chan,ZHANG Qiu-hua and SUN Gui-yuan — 2013/11/15 10:48:51

Objective To investigate the preliminary pharmacological screening of Cassia nomame. Methods The effect of aqueous extract from C. nomame on gastrointestinal motor function was investigated by assessing the intestinal transit rate (ITR) of charcoal modeled into gastrointestinal motility dysfunction (GMD) by the administration of dopamine, atropine, or noradrenaline to the rats, respectively. Diuresis was studied in vivo by estimating the urine output. The anti-inflammatory activity was expressed as the percentage of swelling reduction by comparison on the mean thickness of ear swelling in mice. Results The ITR in these GMD animals was significantly retarded compared to that in normal animals. The retardation, however, was significantly inhibited by the ig administration of C. nomame (2 g/kg) for all GMD animals. The results suggested that C. nomame had the potential for development into a prokinetic agent that could prevent or alleviate GMD in patients. C. nomame increased urine output and suppressed significantly ear swelling induced by dirnethyl benzene in mice. Conclusion C. nomame could increase the gastrointestinal contractile activity of rats and has the effects of diuresis and anti-inflammation.

Biodiversity and Antimicrobial Activity of Endophytic Fungi in Angelica sinensis

JIANG Shu,QIAN Da-wei,YANG Nian-yun,TAO Jin-hua and DUAN Jin-ao — 2013/11/15 10:48:51

Objective To systematically investigate the biodiversity, ecological distribution, and antimicrobial activities of the endophytic fungi in Angelica sinensis growing in several natural habitats in China. Methods The isolation, culture, and identification of microorganism and mycelium growth inhibition test were adopted, and the relative data were analyzed by the statistical methods. Results A total of 206 isolates of endophytic fungi representing 22 species were collected at different time periods from A. sinensis in three locations. Melanconiaceae (45.1%) was the most prevalent followed by Dematiaceae (34.0%), mycelia sterilia (17.0%), Tuberculariaceae (2.9%), Moniliaceae (0.5%), and Leptostromataceae (0.5%). The main genera were Gongromella Ribaldi, Coniosporieae Lk. ex Fr., Fusella Sacc., Myxormia B. ex Br., Ozonium Lk. ex Fr., Pestalotia de Not., Phacodium Pers. ex Wallr., and Sphaceloma de Barry. Endophytes from the samples of Min county showed more diversity, percentage colonization, and species richness compared to other two locations. Endophytic colonization frequency was also greater in Min county (56.5%) than those in Heqing county (29.5%) and Baoxing county (17.0%). As to endophytic community in different plant tissues, the maximum endophytes species richness and diversity appeared in root tissues rather than in stem and leaf tissues from each location. Leaf samples were colonized by greater numbers of endophytes relative to the stem and root samples of the same location. Antimicrobial assay of the 206 endophytes showed that 20% was capable of inhibiting plant pathogenic fungi and eight strains displayed strong inhibition. Conclusion Endophytic fungi isolated from A. sinensis are specific in location, tissue, and season, and diverse in species. They are potential sources of antimicrobial agents which might improve the stress resistance and growth of this medicinal plant and play the important biological functions in ecosystem. Therefore, endophytic fungi will explore a new way to control plant diseases and develop healthful Chinese crude drugs.

Quality Evaluation of Astragali Radix Products by Quantitative Analysis of Multi-components by Single Marker

2013/11/15 10:48:51

Objective To develop a quantitative analysis of multi-components by single-marker (QAMS) method for the simultaneous determination of eight components in Astragali Radix products, and to examine the feasibility of using the method among the different dosage forms and between two different types of compounds. Methods Eight main effective components, campanulin, genistin, ononin, calycosin, genistein, formononetin, methylnissolin, and astragaloside IV were selected as analytes for the quality control of Astragali Radix products. Calycosin was selected as the internal reference substance, the content of which was determined by external standard method; the relative correction factors (RCFs) of campanulin, genistin, ononin, genistein, formononetin, methylnissolin, and astragaloside IV were calculated. In total, twelve Astragali Radix specimen in decoction pieces, as well as in two different dosage forms, such as granule and oral liquid products, were used for the quality control by both methods of external standard and QAMS. The validity of the QAMS method was evaluated by comparison on the quantitative results of the two methods. Results These RCFs were obtained with good reproducibility (RSD < 6.5%) by using ultra high performance liquid chromatography coupled with diode array detector under various chromatographic conditions. Meanwhile, no obvious differences (RSD < 3.98%) were found in the quantitative results of the seven components in twelve samples of Astragali Radix products determined by the two methods. Conclusion QAMS is a reliable and feasible method in determining the components in products of Astragali Radix.

Complementary Application of HSCCC and Semi-preparative HPLC for Rapid Separation of Phenylethanoid Glycosides from Penstemon digitalis

WANG Qi-yan,XIE Jun,FANG Sheng,SHEN Xiu-lian,LI Qi,WU Xiu-hua and TANG Shao-hu — 2013/11/15 10:48:51

Objective Echinacoside and verbascoside are known for their excellent neuroprotective, anti-inflammatory, and anti-oxidative activities, therefore large amount of pure compounds are urgently needed as authentic standards for various in vivo and in vitro studies. Nowadays, they are abundenty extracted from endangered Cistanche spp. Phytochemical studies revealed that Penstemon spp., comprising about 280 species, were rich natural sources for the exploitation and large scale preparation of phenylethanoid glycosides. Thus, rapid isolation and purification of various phenylethanoid glycosides from Penstemon spp. were necessary. Methods The crude extract of P. digitalis was first enriched by AB-8, and then the high-speed counter-current chromatography (HSCCC) combined with semi- preparative high performance liquid chromatography (HPLC) method was adopted to isolate and purify echinacoside and verbascoside. Results Eventually, verbascoside (67.2 mg) with the purity of 92.6% and echinacoside (3.96 mg) with the purity of 98.9% were obtained from 5 g powdered leaves of P. digitalis. Conclusion The present mode of HSCCC coupled with semi-preparative HPLC could be a powerful method for the rapid isolation and purification of other phenylethanoid glycosides.

Simultaneous Determination of Cephaeline and Emetine in Ipecac and Its Preparations Using RP-HPLC

HAN Gui-ru,WANG Yue-feng,FENG Shu-hua and JIA Yong-xin — 2013/11/15 10:48:51

Objective To control the quality of ipecac and its preparations, and to investigate the simultaneous quantitative determination of cephaeline and emetine. Methods After ultrasonic extraction with acidic methanol solution or direct diluting preparations, cephaeline hydrochloride and emetine hydrochloride in ipecac and its preparations were separated within 20 min using a mixture of acetonitrile-methanol-0.1% phosphoric acid (9:3:88) as the mobile phase on a C18 column by HPLC. UV detector was set at 205 nm. The flow rate was set at 1.0 mL/min. Results The methodological study showed that a good linear correlation existed in the range of 0.014 56—0.2184 μg (r = 0.999 97) for cephaeline hydrochloride and 0.0321—0.321 μg (r = 0.999 97) for emetine hydrochloride, respectively. The average recovery of cephaeline hydrochloride and emetine hydrochloride was 96.93% and 99.47%, and the RSD values (n = 9) were 1.31% and 2.02%, respectively. Conclusion The assay is sensitive, accurate, specific, and applicable to comprehensive evaluation on the quality of ipecac and its preparations.

Screening of Phytoestrogenic Effective Extracts and Dose of Cistanche deserticola

LI Wen-lan,CHEN Qiang,YANG Bo,GAO Shang and ZHANG Jin-jin — 2013/11/15 10:48:51

Objective To screen the phytoestrogenic effective extracts and dose of Cistanche deserticola including estrogenic and anti-estrogenic activities. Methods The effect of phytoestrogen was determined through uterus growth test in low and high estrogen female model mice. Then MTT assay of the estrogen-dependent breast cancer cells MCF-7 was conducted with the medicated serum of mice. Results After ig administration with 95% ethanol extract of C. deserticola [EECD, 30 g/(kg·d)], the uterus coefficient of low estrogen model mice increased. The medicated serum of 30 g/(kg·d) EECD significantly promoted the proliferation of MCF-7 cells. 40 g/(kg·d) EECD + diethylstilbestrol significantly inhibited the growth of the uterus in high estrogen model mice and the proliferation of MCF-7 cells as well. Conclusion With the dose of 30 g/(kg·d), EECD could exert quasi estrogen effect, and with the dose of 40 g/(kg·d), EECD could exert the estrogen antagonistic action. The method established is accurate and reliable, which could be used for the follow-up studies on the phytoestrogen material basis of C. deserticola.

Simultaneous Determination of Four Active Ingredients in Wuji Pellet by HPLC

WANG Qun,LIU Wen,YANG Song,CHEN Da-ye and CHEN Zhong-fen — 2013/11/15 10:48:51

Objective To apply the high performance liquid chromatography (HPLC) method to the determination of four ingredients including paeoniflorin, berberine, evodiamine, and rutaecarpin in Wuji Pellet. Methods Using Thermo-ODS2-Hypersil column (250 mm × 4.6 mm, 5 μm) and taking acetonitrile-0.05 mol/L monopotassium phosphate as mobile phase for elution, the conditions were as follows, flow rate of 1.0 mL/min, UV detector wavelength of 225 nm, and column temperature of 25 ℃. Results The standard curves of paeoniflorin, berberine, evodiamine, and rutaecarpine showed a good linear relationship in 0.0124—0.124, 0.0349—0.349 mg/mL, and 0.6—6, 0.5—5 μg/mL. The average recovery rates were 99.45%, 97.69%, 98.18%, and 98.46%, respectively. Conclusion The method is of high sensitivity and nice reproducibility, and it could be used for the simultaneous determination of the four ingredients in Wuji Pellet.

Comparison on HPLC Fingerprints between Fraxini Cortex and Its Eye Drop

XIE Rui-fang,ZHAO Qiu-hua,LI Zhi-cheng and ZHOU Xin — 2013/11/15 10:48:51

Objective To compare the HPLC fingerprints of Fraxini Cortex and its eye drop. Methods Using esculin and esculetin as reference substances, HPLC method was established for fingerprint chromatography of Fraxini Cortex and its eye drop. The similarity was analyzed by similarity evaluation and system cluster analysis. Results For Fraxini Cortex, eight peaks were separated, among which four common peaks were obtained; for eye drop, eleven peaks were separated including six common peaks. Similarities of both fingerprints were good. Cluster analysis further showed that different sources resulted in the variance of ingredients in crude drug. Both materials and eye drop had some common peaks such as esculin and esculetin, on the other hand, they had some different peaks which might be caused by extracting process. Conclusion HPLC fingerprint chromatogram could be applied for the quality control of Fraxini Cortex and its preparations; the pharmaceutical process may be responsible for the variance of ingredients.

Fingerprint Analysis of Codonopsis Radix by HPLC Coupled with Chemometrics Analysis

CHEN Fang-fang,QI Huan-yang and SHI Yan-ping — 2013/11/15 10:48:51

Objective To establish a validated high performance liquid chromatography (HPLC) for the fingerprint analysis of Codonopsis Radix and for the determination of lobetyolin. Methods HPLC coupled with diode array detection method was employed to establish the fingerprint profile and quantitative determination of lobetyolin in Codonopsis Radix. Principal component analysis method was employed to analyze the 52 Codonopsis Radix samples. Results The reference chromatogram was generated with 25 common peaks showing good separation from adjacent peaks. Conclusion Statistical analysis of the obtained data demonstrates that the developed HPLC fingerprint combined with chemometric is a reliable method for the similar evaluation and quality assessment of Codonopsis Radix, and other traditional Chinese herbs.

Chemical Constituents from Dried Aerial Parts of Eclipta prostrata

HAN Li-feng,ZHAO Jing,ZHANG Yi,KOJO Agyemang,LIU Er-wei and WANG Tao — 2013/11/15 10:48:51

Objective To study the constituents in the dried aerial parts of Eclipta prostrata. Methods The constituents were isolated and purified by column chromatography and their structures were elucidated by spectroscopic methods (1D, 2D NMR, UV, IR, and HRESI-TOF-MS) and chemical analyses. Results Eight compounds were isolated and identified as 7-O-methylorobol-4′-O-β-D-glucopyranoside (1), 3′-hydroxybiochanin A (2), echinocystic acid 28-O-β-D-glucopyranoside (3), ecliptasaponin A (4), eclalbasaponin I (5), eclalbasaponin IV (6), echinocystic acid (7), and 3-oxo-16α-hydroxy-olean-12-en-28-oic acid (8). Conclusion Compound 1 is a new compound and compound 3 is obtained from this genus for the first time.

Sesquiterpenes from Stems of Syringa pinnatifolia var. alashanensis

WANG Qing-hu,AO Wu-li-ji,WU Xiao-lan,TAI Wen-quan and DAI Nayintai — 2013/11/15 10:48:51

Objective To study the chemical constituents of the volatile oil from the stems of Syringa pinnatifolia var. alashanensis. Methods The volatile oil from the stems of S. pinnatifolia var. alashanensis was obtained by steam distillation. They were isolated by chromatography on silica gel, Sephadex LH-20 columns and TLC, etc. The structures were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques. Results Five sesquiterpenes were isolated from the volatile oil. Conclusion Compound 1 is a new sesquiterpene.

A New Diarylheptanoid from Barks of Mangifera indica

YU Xiao-xia,XIAO Yue,XU Zu-jiang and LI Guo-cheng — 2013/11/15 10:48:51

Objective To study the chemical constituents from the barks of Mangifera indica. Methods The constituents were separated and purified by different methods of chromatography, and their structures were elucidated by IR, MS, 1D and 2D NMR techniques. Results Six compounds were isolated from the barks of M. indica. Their structures were identified as mangiferone (1), mangiferin (2), myricetin (3), myricitrin (4), rutin (5), and quercetin (6). Conclusion Mangiferone (1) is a new diarylheptanoid compound isolated from the barks of M. indica.

Cloning and Expression Analysis on 3-Hydroxy-3-methyl-glutaryl-coenzyme A Reductase from Aquilaria sinensis

2013/8/17 18:57:33

Objective To clone the full-length cDNA of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) from Aquilaria sinensis (AsHMGR1) and to analyze its expression profile in different tissues and in response to different treatments. HMGR is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate pathway. Methods RT-PCR and RACE were used to clone the full-length cDNA of HMGR from A. sinensis based on the conserved HMGR gene fragments. The bioinformatic analysis was performed on its nucleic acid and protein sequence. The expression profile of AsHMGR1 in different tissues and in response to different treatments was analyzed by quantitative RT-PCR. Results The full-length AsHMGR1 cDNA was 2026 bp, containing a 1719 bp open reading frame which encoded a protein of 572 amino acids. Amino acid sequence homology alignment and phylogenetic analysis demonstrated that AsHMGR1 belonged to the HMGR gene family. The detection of tissue expression patterns showed that AsHMGR1 was mainly expressed in the stem, followed by roots and branches. AsHMGR1 could be stimulated by methyl jasmonate and H2O2 to varying degrees in a time-dependent manner. Conclusion These data will provide a foundation for further investigation on AsHMGR1 functions and regulatory mechanisms in sesquiterpene synthesis in A. sinensis.

Toxicity of Danshen Injection in Beagle’s Dogs by Repeated iv Injection

2013/8/17 18:57:33

Objective To re-evaluate the potential toxicity of Danshen Injection (DI) in Beagle’s dogs by repeated iv injection. Methods DI was iv given to the dogs at the doses of 0, 1.6, 5.4, and 16.0 g/(kg·d) (4 per sex per group) for 13 weeks. During the test period, the clinical signs, mortality, body weights, food consumption, rectal temperature, ophthalmoscopy, electrocardiography, urinalysis, hematology, serum biochemistry, organ weights, gross findings, and histopathology were examined. Results Dogs iv given with DI at the doses of 0, 1.6, 5.4, and 16.0 g/(kg·d) for 13 weeks had no drug-related changes in mortality, body weight, food consumption, temperature, electrocardio- graphy, ophthalmoscopy, urinalysis parameters, and organ weights. The hematological parameter data showed a significant decrease in red blood cells and hemoglobin concentration in the high-dose group and a significant increase in activated partial thromboplastin time suggesting an effect on haemopoiesis. For biochemical parameters, a significant decrease in glucose and a significant increase in total bilirubin were observed in the high-dose group, and the latter was considered to be toxicologically insignificant as lack of histopathological correlate. However, the histopathological examinations of the injection site showed that DI could cause dose-dependent focal inflammation. Conclusion That the iv injection with DI into dogs at 16 g/(kg·d) for 13 weeks could cause the decreases in red blood cell parameters and glucose, as well as the lesions of the injection site. The no observed adverse effect level is 5.4 g/(kg·d), which suggests that safe clinical dosing be possible.

Prevention of Cistanche salsa Extract on Hepatic Fibrosis Induced by Carbon Tetrachloride in Rats

YANG Feng-rui,WEN Du-su,FANG Bu-wu,LOU Jian-shi and MENG Lin — 2013/8/17 18:57:33

Objective To explore the antifibrotic effect of echinacoside on carbon tetrachloride (CCl4)-induced hepatic fibrosis in rats. Methods Male Wistar rats were randomly divided into normal control (n = 8), model (n = 14), and echinacoside treatment (n = 14) groups. The hepatic fibrosis model was induced by CCl4 compositor. The rats were ig administered with echinacoside at a daily dose of 50 mg/kg. The anti-oxidant status, liver function parameters, and hepatic hydroxyproline content were detected by chromatometry. The serum levels of hyaluronic acid (HA), type IV collagen (CIV), type III precollagen (PIIIP), and laminin (LN) were assayed with radioimmunoassay. The hpatic injury was detected by haematoxylin-eosine staining. The deposition of collagen was observed with Masson staining. Results Echinacoside increased the superoxide dismutase activity and reduced the levels of malondialdehyde, aspartate aminotransferase, alanine aminotransferase, HA, CIV, PIIIP, and LN in serum. Echinacoside could also reduce the hydroxyproline content in liver, alleviate hepatic injury, and inhibit collagen deposition. Conclusion Echinacoside possesses antihepatic fibrosis effect.

Protection of Citrullus colocynthis Fruit Extracts on Carbon Tetrachloride-induced and Bacillus Calmette-Guerin plus Lipopolysaccharide-induced Hepatotoxicity in Mice

2013/8/17 18:57:34

Objective To investigate the hepatoprotective activities of the extracts from Citrullus colocynthis (ECC), a native plant used as traditional Uigur Medicine on acute liver injury in mice. Methods The activities of ECC of petroleum ether (ECCPE), chloroform (ECCC), ethyl acetate (ECCEA), n-butyl alcohol (ECCBA), and water (ECCW) were evaluated in vivo using two experimental models, carbon tetrachloride (CCl4)- and bacillus calmette-guerin (BCG) plus lipopolysaccharide (LPS)-induced acute hepatotoxicity in mice. The contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum were determined and the liver histological examination was carried out, respectively. Results The pretreatment with ECC for 7 d obviously reduced the impact of CCl4 toxicity on the serum markers of liver damage, ECCEA and ECCC with a significant difference of AST (P < 0.01, 0.05, respectively) and ALT (P < 0.05, 0.01, respectively). The protective activity was reconfirmed against BCG + LPS-induced injury and the serum enzymatic levels were obviously elevated, for ECCEA and ECCC with a significant difference of AST (P < 0.05, 0.01, respectively) and ALT (P < 0.01, 0.05, respectively). Conclusion That ECCEA and ECCC are the potent hepatoprotective extracts that could protect liver against the acute injury, and this ability might be attributed to their hepatoprotective potentials.

Simultaneous Determination of Eight Bioactive Constituents in Shensong Yangxin Capsule by UPLC

JIANG Xin-gang,JIA Ji-ming and LI Ye-shuang — 2013/8/17 18:57:34

Objective To develop a ultra performance liquid chromatography (UPLC) method for the quality evaluation of Shensong Yangxin Capsule (SYC). Methods The Waters Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) was used. Acetonitrile and water containing 0.1% phosphoric acid were used as mobile phases of UPLC with gradient elution. The detection wavelengths were set at 203 (ginsenoside Rb1), 286 (salvianolic acid B), 230 (paeoniflorin), 221 (schisantherin A), 280 (sodium danshensu), 327 (chlorognenic acid), 335 (spinosin), and 345 nm (berberine hydrochloride), respectively. The flow rate was set at 0.4 mL/min and column temperature was 35 ℃. Results The contents of paeoniflorin, salvianolic acid B, schisantherin A, sodium danshensu, chlorognenic acid, spinosin, berberine hydrochloride, and ginsenoside Rb1 were determined from 10 batches of SYC. Conclusion The method of the quality evaluation of SYC has acceptable precision, reproducibility, and stability, which could be used as a new method for the quality control of SYC.

Optimization for Preparation of Lemon-catalyzed Ginsenoside Rg3 by Response Surface Method

SUN Cheng-peng,GAO Wei-ping,ZHAO Bao-zhong and CHENG Le-qin — 2013/8/17 18:57:34

Objective Ginsenoside Rg3 was prepared by converting protopanaxadiol (PPD)-type saponins using lemon juice as the catalyst. Methods Preparation of ginsenoside Rg3 was optimized by response surface method (RSM) based on a four-factor and five-level central composite design. Results The optimal yield of ginsenoside Rg3 was predicted to be 75.98% in the combination of the factors (PPD-type saponins concentration of 23.6 mg/mL, lemon juice concentration of 97.6% at 85.7 ℃ for 130.0 min) through the canonical analysis and ridge analysis with maximum responses. Under the optimum conditions, the actual yield of ginsenoside Rg3 was 75.57%. Conclusion RSM is effective to optimize the preparation of ginsenoside Rg3 by lemon-catalyzed PPD-type saponins. The achievement for the preparation of ginsenoside Rg3 would promote the development of health-care production.

A Rapid and Effective HPLC Method for Identification of Bovis Calculus in Chinese Patent Medicines

XIA Jing,XIA Li,WANG Wei-yi,SHA Yi-wei,WANG Ke and JI Shen — 2013/8/17 18:57:34

Objective Due to the limited resource and the large demand, many kinds of Bovis Calculus (BC) including artificial Bovis Calculus (ABC), in vivo cultured Bovis Calculus (in vivo CBC), and in vitro cultured Bovis Calculus (in vitro CBC) were used in Chinese patent medicines (CPMs). Previous studies have shown that the chemical constituents of ABC and their properties were different from other BC. The two types of CBC with much higher price than ABC were approximately equivalent with natural Bovis Calculus in quality and clinical effect. The aim of the study is to establish a rapid and effective method for the identification of BC in CPMs. Methods An HPLC method with the higher specificity for analyzing bilirubin was established to distinguish ABC from other three kinds of BC by comparing the change of bilirubin content with the addition of EDTA-2Na as the extraction solvent and stabilizer. Results The bilirubin content in CPMs containing ABC was basically unchanged, while that in CPMs containing other kinds of BC showed significant difference. The proposed method was employed to analyze a variety of CPMs containing Bovis Calculus (CPMBCs) and proven to be universal. Conclusion An effective analytical method is established for the quality control of CPMBCs and further ensures the safety and efficacy of these drugs in clinical practice.

A New Cucurbitane Triterpene in Acid-treated Ethanol Extract from Momordica charantia

CAO Jia-qing,ZHANG Bo-yu and ZHAO Yu-qing — 2013/8/17 18:57:34

Objective To study the chemical constituents in the acid-hydrolyzed ethanol extract from Momordica charantia. Methods The ethanol extract from M. charantia was hydrolyzed by 36% HCl and the hydrolysate was isolated by silica gel column chromatography and preparative HPLC. The structures of the isolated compounds were identified by spectral analyses, physical constants, and chemical evidences. Results Two cucurbitane triterpenoids were isolated and identified as 5β,19-epoxy-cucurbita-6,22E,24-trien-3β-ol (1) and cucurbita-6,22(E),24-trien-3β-ol- 19,5β-olide (2). Conclusion Compound 1 is a new compound.

A New Secoiridoid Glycoside from Roots of Picrorhiza scrophulariiflora

2013/8/17 18:57:34

Objective To study the chemical constituents in the roots of Picrorhiza scrophulariiflora. Methods The chemical constituents in the roots of P. scrophulariiflora were separated and purified with chromatographic methods. The structures were elucidated by spectroscopic methods and chemical analyses. Results A new secoiridoid glycoside, picrogentioside II (1) was successfully isolated from the roots of P. scrophulariiflora. Conclusion Compound 1 is a new secoiridoid glycoside.

Phenolic Compounds from Roots of Imperata cylindrica var. major

LIU Rong-hua,CHEN Shi-sheng,REN Gang,SHAO Feng and HUANG Hui-lian — 2013/8/17 18:57:34

Objective To study the chemical constituents from the roots of Imperata cylindrica var. major. Methods The chemical constituents were isolated and purified by combination of silica gel, Sephadex LH-20, polyamide, and ODS column chromatography. The structures of the isolated compounds were identified by means of spectral data and physicochemical property. Results Twelve phenolic compounds were isolated from I. cylindrica var. major and identified as 4,7-dimethoxy-5-methylcoumarin (1), 7-hydroxy-4-methoxy-5-methylcoumarin (2), 7-O-β-D-glucopyranosyl- 4-methoxy-5-methylcoumarin (3), 6-hydroxy-5-methoxyflavone (4), 5-methoxyflavone (5), 5,7-dihydroxy-8- methoxyflavone (6), 4-hydroxybenzaldehyde (7), 4-hydroxy-cinnamic acid (8), 4-hydroxy-3-methoxybenzoic acid (9), 3,4-dimethoxyphenyl-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (10), impecyloside (11), and graminone A (12). Conclusion Compounds 2—4, 6, and 10 are obtained from this plant for the first time.

A Novel Bihomoflavanonol with an Unprecedented Skeleton from Pteridium aquilinum

CHEN Nai-dong,CHEN Nai-fu,CHEN Cun-wu and ZHANG Li

Objective To seek the flavonoids with the unique structure and to investigate the chemical ingredients in the flavonoid-rich plant—Pteridium aquilinum. Methods The 80% EthOH extract from the degreased powder of P. aquilinum was partitioned by petroleum ether, CHCl3, EtOAc, n-butanol, and water, respectively. The EtOAc fraction was sequentially subjected to silica gel column, repeated Sephadex LH-20 column, and preparative TLC to give a new compound. The antitumor activity of the novel flavonoid was primarily evaluated by MTT. Results Compound 1, a biflavonoid with the unique structure named as pteridium III with an unprecedented bihomoflavanonol skeleton, was isolated from P. aquilinum. Compound 1 showed the in vitro antitumor activity against lung cancer cell NCI-H46, melanoma cell A375, and glioma cell U-7MG corresponding to the IC50 values of 22.9, 106.7, and 1540.5 μmol/L, respectively. No inhibition on gastric carcinoma SGC-7901 and prostatic carcinoma PC-3 was observed in the experiment. Conclusion A rare bihomoflavononol derivative, pteridium III, is obtained from the plant, which could enrich our knowedge on the chemical structures of flavonoids and bioactive constituents in P. aquilinum.

A New Prenylated Xanthone from Root Barks of Cudrania cochinchinensis

LIU Zhi-ping,WEI Wan-xing,ZHOU Min,GAN Chun-fang and LIU Sheng

Objective To study the chemical constituents from the root barks of Cudrania cochinchinensis. Methods The chemical constituents were isolated and purified by silica gel column chromatography. The structures of the compounds were identified on the basis of spectral data (MS, 1H-NMR, 13C-NMR, and 2D NMR) and by the comparison of spectroscopic data with the reported values in the literatures. Results A new xanthone, 1,6,7-trihydroxy-4-(1,1- dimethylallyl)-3-methoxyxanthone (1) and a known prenylated xanthone 1,5,6-trihydroxy-4-(1,1-dimethylallyl)-3- methoxyxanthone (isocudraniaxanthone B, 2) were isolated from the root barks of C. cochinchinensis. Conclusion Compound 1 is a new prenylated xanthone. Isomers 1 and 2 are obtained from this plant for the first time.

Chemical Constituents in Essential Oils from Elsholtzia ciliata and Their Antimicrobial Activities

TIAN Guang-hui

Objective To compare the chemical constituents in the essential oils from the leaves, flowers, and seeds of Elsholtzia ciliata and their antimicrobial activities. Methods The chemical constituents in essential oils were extracted by the hydro-distillation method and analyzed by GC-MS. The chemical constituents in essential oils were identified on the basis of comparison on their retention indices and MS spectrum with published data. Moreover, the antimicrobial activities of the chemical constituents in the oils against the growth of six bacteria strains and one pathogenic yeast strain were evaluated by using minimum inhibitory concentration and minimum bactericidal concentration methods. Results A total of 58 compounds were identified, while compounds 31, 35, and 36 were identified in the essential oils from the leaves, flowers, and seeds, respectively. Fifteen compounds were identified as shared constituents in the leaves, flowers, and seeds. The chemical constituents in the essential oils showed the inhibitory activities against the six bacteria strains and the yeast strain. Conclusion The major constituents are different in the essential oils of the leaves, flowers, and seeds. The major chemical constituents in the essential oils are monoterpenoids and sesquiterpenoids. And the chemical constituents in the essential oils obtained from the leaves show higher inhibitory activities especially against Bacillus subtillis CMCC63501 and Escherichia coli ATCC25922.

Pharmacology and Toxicology of Extract from Arcangelisia gusanlung

HU Hai-zhen,DONG Zhi,ZHU Yi and CHEN Guo-biao

Objective To study the pharmacology and toxicology of the extracts from Arcangelisia gusanlung (EAG). Methods The anti-inflammatory activities were investigated using various inflammatory models including ear edema induced by xylene in mice, paw edema induced by carrageenan, and cotton pellet granuloma in rats. The analgesic effect was observed in hot-plate test and writhing test in mice and the antipyretic effect was observed in rat fever model induced by yeast. The antitussive action was tested in mice by sequential method and expectorant action was evaluated by tracheal excretion of phenol red. The antidiarrhea function was observed on normal intestinal propulsion of mouse model of diarrhea induced by decoction of Sennae Folium. The toxicity was measured by toxicological experiment. Results Each dose of EAG could significantly inhibit the paw edema, cotton pellet granuloma, and intestinal propulsion. EAG significantly reduced writhing times and amount of wet manure. Obvious antipyretic action to fevered rat was observed. EAG obviously increased the tracheal excretion of phenol red and prolonged the latency of cough. No toxic reaction was shown in the observed period, and the maximum tolerance dose of mice was equivalent to 1360 times of common-used dose in human. Conclusion The clinical dosage of EAG is safe, and its anti-inflammatory, analgesia, antipyresis, antitussive, expectorant, and antidiarrhea effects are significant

Pharmacokinetics and Tissue Distribution of Orientin from Trollius chinensis in Rabbits

TAN Xiao-hong,QU Cai-hong,WANG Shu-hua and AN Fang

Objective To study the pharmacokinetics (PK) and tissue distribution of orientin in rabbits. Methods The high-performance liquid chromatography method was developed and validated for the determination of orientin in rabbit plasma and tissues. The PK characteristics of orientin in rabbits were compared by three kinds of administration, iv, im, and ip injection, using 3P97 pharmacokinetic software to examine the PK parameters. Results The elimination half-life (t1/2β) of three kinds of administration was higher than the distribution half-life (t1/2α). The PK of orientin eliminating in rabbit fitted the two compartment model. Orientin was mainly eliminated in rabbit. The distribution of orientin in tissue after iv injection was investigated, and the order of orientin AUC in each group was kidney > liver > lung > spleen > heart > brain. Conclusion Orientin is distributed extensively in tissues such as kidney, liver, and lung. It is difficult for orientin to cross the blood brain barrier.

Immunological Effects of Total Flavones from Leaves of Choerospondias axillaris on Mice

LIU Xin-yuan,YANG Yu-mei,HAN Feng,ZHANG Hao-nan,DU Jian-xi and HAO Xin-min

Objective To investigate the effect of total flavones from the leaves of Choerospondias axillaris (TFLCA) on the immune function of normal mice and to provide the experimental basis for the reasonable application of C. axillaris. Methods The carbon clearance method, cutaneous delayed hypersensitivity reaction method, serum hemolysin method, and index of immune organs were used to study the effect of TFLCA on the immune function of mice. Results TFLCA could enhance the phagocytic function of mononuclear macrophage and the cutaneous delayed hypersensitivity reaction of mice, and increase the content of hemolysin antibody and the thymus index in mice. Conclusion TFLCA could improve the celiac macrophage activity and specific immunity of mice, and TFLCA, consisting with the total flavones of Choerospondiatis Fructus (TFCF), has the effect on the immune function of mice. So both TFLCA and TFCF have the regulatory effects on the immune function of mice.

Anticancer Effects of Zaoxiu Compound Decoction on H22 Mice with Heps

JIANG Yan-qing,ZHANG Wen-yun and YAN Sheng-ying

Objective To investigate the anticancer effects of the Zaoxiu Compound Decoction (ZCD) on H22 mice with Heps and the influence on the construction of H22 cells. Methods With in vivo techniques, the antineoplastic drugs were given to hepatoma H22 model mice which were randomly divided into negative control (physiological saline), positive control (cyclophosphamide), high- and low-dose ZCD groups. After 10-d administration, the pathological examinations of tumor tissue, livers, and kidneys in mice were carried out and the inhibitory rate on tumor was calculated. Results The high- and low-dose ZCD had obvious inhibition on H22 cells. The pathological observation showed that high- and low-dose ZCD had a strong inhibitory effect on H22 cells as well as positive control, with few toxic effects to the livers and kidneys in mice. Conclusion ZCD has the anticancer effect and has no obvious toxic effects on the model mice.

Effect of Jiaosanxian on Reducing Blood Lipid

YIN Ai-wu,TIAN Run and ZHAO Jiao

Objective To investigate the effects of Jiaosanxian (JSX, consisted of stir-baked Hordei Fructus Germinatus- Crataegi Fructus-Massa Fermentata Medicinalis) on reducing blood lipid. Methods The model of hyperlipidemic mice was established by feeding high-fat diet. Kunming mice were randomly divided into six groups: blank, hyperlipidemic, Zhibituo, low-, mid-, and high-dose [100, 200, and 400 mg/(kg·d)] JSX groups, and were continuously ig administered for 28 d. The contents of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) in serum and liver and liver index were determined. Results The contents of TC, TG, and LDL-C in serum or liver were decreased significantly but the contents of HDL-C were increased more significantly in the mid- and high-dose JSX groups than those in hyperlipemic group. The weight and liver index were decreased significantly with the dose increasing of JSX. The lipid-decreasing effects were improved with the dose increasing. Conclusion These results suggest that JSX has the significant effects on hyperlipidemia. It could provide the experimental basis for the clinical use of JSX for the treatment of hyperlipidemia.

A New Procedure for Separation and Purification of Scutellarin from Erigeron multiradiatus

ZHANG Zhi-feng,LU Lu-yang and LIU Yuan

Objective To establish a new procedure for isolating scutellarin from Erigeron multiradiatus. Methods A proposed method was developed by combining macroporous resins with ODS column. Firstly, E. multiradiatus was extracted by ultrasound with 80% methanol. Preliminary separation was performed on macroporous resin column. The performance and adsorption characteristics of three macroporous resins, D140, D141, and D605, were compared and the enrichment procedure was optimized. Further purification was carried out by medium pressure liquid chromatography (MPLC) with ODS column. Results It was demonstrated that D141 had better extractive effects on scutellarin. The MPLC conditions were optimized as follows: 15% ethanol aqueous as mobile phase with flow rate at 2.5 mL/min. The yield and purity of the isolated scutellarin were 1.20 mg/g and 96.5%, respectively. Conclusion The overall procedure is efficient and low-cost, which is considered suitable for the separation and purification of scutellarin from E. multiradiatus. The results provide the scientific basis for developing and using scutellarin in clinic.

Optimization of Jiawei Qing’e Oral Fast Disintegrating Tablets Based on Response Surface-Central Composite Design

Objective To apply the response surface-central composite design to developing and optimizing the oral fast disintegrating tablets (ODT) formulation for Jiawei Qing’e, a kind of prescription of Chinese herbal medicine. Methods The bitterness of Jiawei Qing’e was masked using Eudragit E-100 by solvent evaporation technique. Response surface approach was applied to investigating the interaction of formulation parameters in optimizing the formulation. The independent variables were Eudragit E-100/drug ratio (X1), amount of disintegrants (X2), and the amount of diluents (X3). The disintegration time (Y1), hardness (Y2), and weight variations of the tablets were characterized. Results The models predicted levels of X1 = 4.63%, X2 = 5.25%, and X3 = 34.33%, for the optimal formulation having a hardness of 3.0 kg with the disintegration time of 30 s within experimental region. The observed response of Y1 = 26.5 s and Y2 = 3.14 kg reasonably agreed with the predicted response. Conclusion Response surface methodology shows the good predictability and reliability in optimizing the formulation. The optimized ODT of Jiawei Qing’e has acceptable taste, rapid disintegrating ability, and good mechanical strength.

Seasonal Variation, Microscopic and Chromatographic Analysis of Leaves in Malus hupehensis: A Protocol for Its Quality Control

SHEN Tao,XIANG Lan,REN Dong-mei,WANG Shu-qi,YANG Ming-ren and LOU Hong-xiang

Objective To establish a quality control protocol based on microscopic, TLC, and HPLC methods, and to verify the optimal harvesting time for the leaves of Malus hupehensis (LMH). Methods The LMH were pulverized into powder for microscopic identification or TLC and HPLC analysis after ultrasonic extraction with methanol. Seasonal variations of the phlorizin content and average leaf weight were determined by HPLC analysis and weighing up the leaves collected from May to October. Results Microscopic and macromorphologic characteristics have been described for the leaf identification. A qualitative TLC assay and a quantitative HPLC method have been established for the quality control of LMH. Phlorizin was selected as a reference marker, which resolved at Rf 0.53 in TLC assay and at 14.0 min in HPLC assay. The content of phlorizin decreased gradually from 17.0% in leaves collected in May to 7.5% in October. The average leaf weight reached the level of 0.6 g in August and maintained until its falling. Conclusion These methods are simple, selective, accurate, and reliable for the quality control of LMH. The period from late August to early September is suggested as the optimal harvesting time of the LMH.

Effects of Postharvest Processing and Geographical Source on Phytochemical Variation of Corydalis Rhizoma

ZHAO Yun-peng,LI Jing-hui,YANG Shu-ting,FAN Jie and FU Cheng-xin

Objective To assess the relative contributions of postharvest processing and geographical source to phytochemical variation of Corydalis Rhizoma, and rhizome of Corydalis yanhusuo, and to examine what phytochemical components are the most sensitive to the differences of each factor and how they change. Methods HPLC fingerprinting and LC-MS coupled with chemometric approaches were applied. Results The results of principal component analysis (PCA) and hierarchical cluster analysis (HCA) explicitly demonstrated the postharvest processing could produce a greater impact on the phytochemical profiles of Corydalis Rhizoma than geographical source. The contents of most compounds increased after water boiling while decreased after sulphur-fumigation. Protopine, coptisine, and palmatine were the most variable components in processing. Geographical sources also led to a remarkable phytochemical differentiation, in which the environmental variation of the three regions might play a role. Dehydrocorybulbine, coptisine, dehydrocorydaline, and protopine varied most among the three production regions and decreased sequentially in Zhejiang, Shaanxi, and Jiangsu provinces, China. Conclusion Both postharvest processing and geographical source should be enhanced with the priority for the former in the quality control of Corydalis Rhizoma. The application of boiling is supported but the consistency should be improved in practice. Sulphur-fumigation is strongly suggested to be abandoned.

Preparation and Crystal Structure of Acetyl Hemerocallin and Structural Revision of Hemerocallin

LIU Yan-ze,WU Jie,ZHU Yu,DU Chen-xia,CHANG Jun-biao and XIE Jing-xi

Objective To revise the structure of hemerocallin, an active and toxic compound isolated from the roots of Hemerocallis fulva, which was first reported in 1974 based on the chemical and spectral data. Methods The structure of acetyl hemerocallin was determined through the acetyl derivative preparing and X-ray diffraction. Results The title compound acetyl hemerocallin (1) has been prepared and structurally characterized by X-ray single-crystal diffraction method. X-ray analysis revealed that the two naphthalene rings in compound 1 were significantly twisted with an average dihedral angle of 44.1°. The two acetate groups in each naphthalene ring adopted anti-parallel conformation. In addition, the π-π stacking interactions were found in the crystal structure, the molecules were linked into a supramolecular network, and the crystal structure was stabilized. Conclusion The structure of hemerocallin is identified as 2,2′-bi(1,8-dihydroxy-6-methyl-7-acetylnaphthalene) (3)

Alleviation of PEGylated Puerarin on Erythrocyte Hemolysis Induced by Puerarin in Glucose-6-phosphate Dehydrogenase- deficient Rats

Objective To explore and analyze the reducing hemolytic effects of PEGylated puerarin (PEG-PUE) on erythrocytes induced by PUE in glucose-6-phosphate dehydrogenase (G6PD)-deficient rats. Methods The rat model with G6PD-deficiency was established via sc injecting 1% acetylphenyl-hydrazine. Then the G6PD-deficient erythrocyte suspension obtained from this rat model was used to evaluate the hemolytic effects of PUE and the reducing hemolytic effects of PEG-PUE via hemolytic activity and erythrocyte osmotic fragility assay. Results It was found that PUE could cause a serious hemolysis to the erythrocyte suspension with the increase of drug concentration and the prolongation of drug incubation time, the hemolytic rate of PUE was up to 40%, while the addition of PEG-PUE to the erythrocyte suspension revealed no significant hemolysis. Additionally, the result of erythrocyte osmotic fragility indicated that PEG-PUE exerted a slight effect on the erythrocyte membranes, and the NaCl concentration that induced 50% hemolysis (32 mmol/L) was about one-third PUE. Conclusion These results demonstrate that PEG-PUE could play a significant role in reducing the side effect of hemolysis induced by PUE. The low hemolytic activity of PEG-PUE makes it a favorable candidate for in vivo tests and PEG-PUE could also provide the useful insight for the further formulation development as an innovative drug.

Anti-hepatitis B Virus Activity of 8-epi-Kingiside in Jasminum officinale var. grandiflorum

ZHAO Gui-qin,YIN Zhi-feng,LIU Li-yan,MAO Xiao-xia and SU Zhan-hui

Objective To evaluate the effect of 8-epi-kingiside (8-Epik) derived from the buds of Jasminum officinale var. grandiflorum (JOG) on hepatitis B virus (HBV) replication in HepG2 2.2.15 cell line in vitro and duck hepatitis B virus (DHBV) replication in ducklings in vivo. Methods The concentration of extracellular hepatitis B e antigen and hepatitis B surface antigen (HBsAg) in cell culture medium was determined by ELISA, respectively. The anti-HBV effects of 8-Epik were also demonstrated in the model of DHBV. 8-Epik was ip given (20, 40, and 80 mg/kg, twice daily) to the DHBV-infected ducklings for 10 d. The isotonic saline liquid diet was ip given as negative control and Lamivudine (50 mg/kg, twice daily) was given as positive control. DHBV DNA was measured at days 0 (T0), 5 (T5), 10 (T10), and day 3 after cessation of treatment (P3) by dot blotting. Results 8-Epik effectively blocked HBsAg secretion in HepG2 2.2.15 cells in a dose-dependent manner [IC50 = (19.4 ± 1.04) μg/mL]. 8-Epik (40 or 80 mg/kg, ip, twice daily) also reduced viremia in DHBV-infected ducks. Conclusion Therefore, 8-Epik is warranted as a potential therapeutic agent for HBV infection.

Effect of Qinglongyi Polysaccharides on Complex Mobility of Erythrocytes in S180 Mice

JI Chen-feng,YUE Lei and JI Yu-bin

Objective To study the effect of Qinglongyi polysaccharides (QP) in the exocarp of Juglans mandshurica on the complex mobility of erythrocytes in S180 mice. Methods Erythrocytes were collected and prepared into suspensions, and the complex mobility of cells was measured using high performance capillary electrophoresis (HPCE). Optimized experimental conditions were as follows: 50 cm × 75 μm capillary, buffer for electrophoresis; phosphate solution containing hydroxypropylmethyl cellulose (0.1 mol/L, pH 7.4), injection pressure 3.448 kPa, injection time 10 s, separation voltage 20 kV, and column temperature 25 ℃. Results The migration time of erythrocytes in S180 mice was longer than that in normal mice, which was 18.09 min for the model group and 12.11 min for the control group, and the complex mobility of erythrocytes in S180 mice was lower than that in normal mice, which was 0.92 × 10?4 cm2/(V?s) for the model group and 1.38 × 10?4 cm2/(V?s) for the control group. It was also found that S180 mice treated by QP could shorten the migration time and increase the complex mobility of erythrocytes. Conclusion QP could improve the complex mobility of erythrocytes in S180 mice, and HPCE could be used as a powerful tool for determining the physiological state and functions of erythrocytes.

Protective Effects of Lentinan against T Lymphocytes Injury in Mice under Chronic Radiation Stress

WANG Yong,LI Ming-chun and FU Qing-jie

Objective To study the effects of lentinan (LTN) on mice exposed to chronic radiation. Methods Animals were divided into three groups (n = 10), they were animals exposed to radiation (Rad), normal control animals (Ctr), and irradiated animals treated with LTN (Rad + LTN). Animal model of chronic radiation stress injury was induced by irradiating mice with 60Co γ-ray for 6 weeks from Monday to Friday consecutively. Before radiation, the mice in Rad + LTN group were ip injected with 0.5 mL LTN (0.01 mg/mL), whereas mice in other groups were injected with 0.9% physiological saline. The effects of LTN treatment on irradiated mice were examined by histological analysis on the spleen. The cell numbers and viability of T lymphocytes, which were isolated from the spleen, were determined by Trypan blue staining. Nitric oxide (NO) production and interleukin-2 (IL-2) secretion in T lymphocytes were also measured. Results Chronic radiation significantly reduced the body weights and the spleen and thymus indexes, associated with reduced T lymphocytes viability and functions, and elevated NO production. Treatment with LTN significantly normalized the elevated NO production, and attenuated the negative outcomes resulting from radiation mentioned above. Conclusion The results suggest that radioprotective effect of LTN may be contributed by improved T lymphocytes viability and functions via regulating the NO and IL-2 production in T lymphocytes.

Aqueous Two-phase Systems with Ultrasonic Extraction Used for Extracting Phenolic Compounds from Inonotus obliquus

ZHAO Yan-xia,LIU Yu-bing,LIU Feng and ZHENG Wei-fa

Objective To optimize the extracting technology of assessing the maximum yield of phenolic compounds (PC) from Inonotus obliquus by single factor experiments and orthogonal array design methods through aqueous two-phase systems combined with ultrasonic extraction. Methods The range of the independent variables, namely levels of acetone and ammonium sulfate, and ultrasonic time were identified by a first set of single factor experiments. The actual values of the independent variables coded at four levels and three factors were selected based on the results of the single factor experiments. Subsequently, the levels of acetone and ammonium sulfate, and ultrasonic time were optimized using the orthogonal array method. Results The optimum conditions for the extraction of PC were found to use 7.0 mL acetone, 5.5 mg ammonium sulfate, with ultrasonic time for 5 min. Under these optimized conditions, the experimental maximum yield of PC was 37.8 mg/g, much higher than that of the traditional ultrasonic extraction (UE, 29.0 mg/g). And the PC obtained by this method had stronger anti-oxidative activities than those by traditional UE method. Conclusion These results indicate the suitability of the models developed and the success in optimizing the extraction conditions. This is an economical and efficient method for extracting polyphenols from I. obliquus.

GC-MS Analysis on Volatile Oil from Branches of Picrasma quassioides

HUANG Lin-fang,WANG Zeng-hui and CHEN Shi-lin

Objective To analyze the volatile oil from the branches of Picrasma quassioides. Methods Volatile oil was extracted from the branches of P. quassioides by steam distillation. GC-MS method was used to analyze the components. Results A total of 49 compounds were isolated, and 46 compounds were successfully identified, which represented over 98% of the total oil composition. The major components of the volatile oil in the branches of P. quassioides included caryophyllene (12.83%), 12-oxabicyclo[9.1.0]dodeca-3,7-diene,1,5,5,8-tetramethyl-[1R- (1R*,3E,7E,11-R*)] (12.29%), 1-hexanol (9.96%), naphthalene, 1,2,3,5,6,7,8,8a-octahydro-1,8a-dimethyl-7- (1-methyletheyl)-[1S-(1a,7a,8aa)] (7.32%), aromadendrene oxide-(2) (6.69%), and α-caryophyllene (3.88%). Conclusion The major components in volatile oil are terpenoids, hydroxy compounds, and other acyclic alkane compounds.

Quality Evaluation of Astragali Radix based on DPPH Radical Scavenging Activity and Chemical Analysis

2014/11/12 16:19:17

Objective To assess the quality of Astragali Radix from different areas based on the biological evaluation and chemical analysis. Methods The bioassay method of 1,1- diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity for Astragali Radix was established. The parameters of DPPH assay including sample extraction time, reaction time, repeatability, and stability were detected. Furthermore, a method of HPLC-MS was developed to simultaneously determine calycosin-7-O-glucoside, ononin, formononetin, and astragaloside IV in Astragali Radix samples. And the total flavonoids and total saponins were detected by spectrophotometry. The relationship between DPPH evaluation and chemical analysis was studied by Pearson correlation analysis. Results Twelve batches of Astragali Radix from different origins showed a wide range of DPPH radical scavenging activities (IC50 = 1.395?9.894 μg/mL). Based on DPPH assay, Sample 10 derived from Inner Mongolia Autonomous Region (IC50 = 1.395 μg/mL) showed the best quality of all samples. Chemical analysis showed that different compounds selected as indices would cause different results for quality evaluation. Pearson correlation analysis revealed that the contents of total flavonoids (P = 0.032), calycosin-7-glucoside (P = 0.035), and astragaloside IV (P = 0.010) were positively correlated with DPPH radical scavenging activity. Conclusion Except for chemical analysis, DPPH radical scavenging activity can be used as a good alternative to assess and control the quality of Astragali Radix.

Protection of Shengmai Recipe on Improving Cardiac Function and Attenuating Kidney Injury in Pressure Overload Rats

2014/11/12 16:19:18

Objective Shengmai Recipe (SMR) is a Chinese patent medicine used for the treatment of chronic heart disease. In order to further assess the renal-protective effect against ischemia lesion of SMR, the cardioprotective effect of SMR on pressure overload-induced left ventricular (LV) systolic dysfunction and the potential mechanism on alleviating myocardial damage, myocardial fibrosis, and renal ischemia lesion in chronic heart failure (CHF) rats were investigated. Methods Rats with partially ligated abdominal aorta were randomly divided into model, Sham, and SMR groups. One week after recovery from surgery, animals were preventively ig administered with SMR at the dose of 810 mg/kg once daily for 8 weeks. Cardiac function and structure, endogenous biomarkers (CK-MB and LDH), myocardial fibrosis, and organ pathological change were estimated by echocardiography, immunodepression and velocity method, hematoxylin- eosin staining, and masson’s trichrome staining, respectively. Results The administration of SMR significantly decreased serum CK-MB and LDH levels and reduced myocardial fibrosis. Interestingly, SMR not only improved cardiac function but also ameliorated kidney injury induced by ischemia in CHF rats. Conclusion SMR could enhance the LV contractile function, reduce myocardial necrosis, and reverse LV remodeling in CHF rats, and most importantly, SMR could be used to treat the renal ischemia injury in pressure overload rats.

Effects of Soothing Liver and Invigorating Spleen Recipe on Lipid Metabolism Disorders in Kupffer Cells of NAFLD Rats by LXRα/SREBP-1c Signal Pathway

2014/11/12 16:19:18

Objective To investigate the mechanism of lipid metabolism disorders in Kupffer cells (KCs) of non-alcoholic fatty liver disease (NAFLD) rats mediated by LXRα-SREBP-1c pathway and the interference of soothing liver and invigorating spleen recipe (SLISR) on it. Methods SD male rats were randomly divided into five groups: normal, model, soothing liver recipe (SLR), invigorating spleen recipe (ISR), and soothing liver and invigorating spleen recipe (SLISR) groups. The rats in treatment groups were administered for 8 weeks. The liver tissue was stained with H&E and oil red O. The levels of hepatic lipid and blood lipid were measured by biochemical analyzer. KCs were isolated from the livers of rats to evaluate the expression of LXRα, SREBP-1C, and FAS mRNA by real-time fluorescence quantitative PCR tests; LXRα, SREBP-1C, and FAS proteins were measured by Western blotting. Results The H&E and oil red O staining results showed that the model rats successfully reproduced typical pathogenetic and histopathological features of NAFLD. The levels of hepatic lipid and blood lipid in the model rats were dramatically increased. Compared with the model group, the values of hepatic lipid and blood lipid in the treatment groups were significantly ameliorated (P < 0.05, 0.01). The yields of purified KCs from each rat were 2×107-3×107. The viability of KCs was higher than 95%, with the purity over 90.18%. Compared with the model group, the expression of LXRα, SREBP-1C, and FAS mRNA and proteins was decreased in all treatment groups, especially in the SLR group (P < 0.05). Conclusion SLISR may protect liver against injury included by lipid metabolism disorders in KCs through LXRα/ SREBP-1c signaling pathway, which may be an important mechanism for the prevention and treatment of NAFLD.

Cost-effectiveness Assessment of Commonly-used Drugs for Hepatoprotection and Enzymes Reduction Based on Decision Tree and Multi-utility Theory

Jia-qi Xu,Rui-fang Xie,Zhi-cheng Li,Jing-yi Tang and Xin Zhou — 2014/11/12 16:19:18

Objective Drugs for hepatoprotection and enzymes reduction are widely used in China but their economic analysis has been ignored in a rather long period of time. A suitable protocol for hepatoprotection and enzymes reduction was recommended in Longhua Hospital. Methods This study was conducted as a retrospective piece. Three therapeutic protocols (compound glycyrrhizic glycoside combined with aspartic ornithine injection, compound glycyrrhizic glycoside combined with phospha- tidylcholine, and compound glycyrrhizic glycoside combined with tiopronin) were selected. Seventy inpatient cases from January 2011 to February 2012 were enrolled and divided into three groups according to different regimens. The cost effectiveness of the three groups was respectively evaluated by incremental cost-effectiveness ratios (ICERs). A decision tree model and multi attribution utility theory were also adopted to analyze the data. Results All the three regimens exhibited good effects on protecting liver functions and reducing the levels of enzymes. Among them, the protocol of compound glycyrrhizic glycoside combined with tioproni expressed the least ICER, the lowest cost but the highest score in the multi-utility. Conclusion The therapeutic method of compound glycyrrhizic glycoside combined with tiopronin is the most cost-effective option in this study.

Influence of Allelochemicals on Microbial Community in Ginseng Cultivating Soil

Yong Li,Yi-xin Ying,Dong-yue Zhao and Wan-long Ding — 2014/11/12 16:19:18

Objective To study the influence of allelochemicals such as benzoic acid, diisobutyl phthalate, diisobutyl succinate, palmitic acid, and 2,2-bis-(4-hydroxyphenyl) propane on the microbial community of ginseng cultivating soil. Methods Soil samples were sprayed with five allelochemicals (100 mg/L) and their mixture (20 mg/L), respectively. Carbon metabolic ability variances were analyzed by Biolog method and genetic polymorphism variance was analyzed by RAPD method. The Nei’s genetic diversity index and Shannon’s information index were calculated. Results Significant differences in carbon metabolic activity were found between allelochemical-treated soils and control. Further analysis indicated that although the carbon-utilizing ability and genetic polymorphism of soils treated with di-isobutyl phthalate, di-isobutyl succinate, and allelochemical mixtures were lower than those of the other treatments, genetic similarities of soils treated with di-isobutyl phthalate, diisobutyl succinate, and allelochemical mixtures were much higher than those of the other treatments. Conclusion Allelochemicals significantly declined the genetic diversity and carbon metabolic activity of microorganisms in newly reclaimed forest soil for ginseng cultivation.

C21 Steroidal Glycosides from Acidic Hydrolysate of Cynanchum otophyllum

Yi-bin Zhao,Quan-shui Fan,Gui-li Xu and Xiao-jiang Hao — 2014/11/12 16:19:18

Objective To investigate the structures of compounds in the rhizome of Cynanchum otophyllum (Asclepiadaceae), and to find new C21 steroidal glycosides. Methods The ethyl acetate extract from the rhizome was subjected to acidic hydrolysis and isolated by column chromatography; The structures of the purified compounds were determined by spectral methods. Literature search confirmed whether those compounds were of new structures. Results Three compounds were isolated and their structures were deacetylmetaplexigenin 3-O-β-D-oleandropyranosyl-(1→4)-α-D-oleandropyranosyl- (1→4)-α-D-oleandropyranoside (1), deacetylmetaplexigenin 3-O-α-D-oleandropyranosyl- (1→4)-β-D-thevetopyranosyl-(1→4)-α-D-oleandropyranoside (2), and deacetylmeta- plexigenin 3-O-β-D-cymaropyranosyl-(1→4)-α-D-oleandropyranoside (3), respectively. Conclusion Compounds 1?3 are new compounds.

Chemical Constituents with Antihyperlipidemic Activities from Desmodium triquetrum

2014/11/12 16:19:18

Objective To study the chemical constituents from Desmodium triquetrum and their antihyperlipidemic activities. Methods The constituents of D. triquetrum were isolated and purified using various column chromatographies. Their chemical structures were elucidated using extensive spectroscopic methods. The lipid-lowering effects of the isolates were evaluated in HepG2 cells. Results Nine compounds were obtained from the ethanol extract of D. triquetrum and determined to be 6′-O-cis-p-coumaroyl- 3,5-dihydroxyphenyl-β-D-glucopyranoside (1), tadehaginoside (2), rutin (3), quercetin-3-O-β-D-glucopyranoside (4), quercetin-3-O-β-D-galactopyranoside (5), 6-O-(E )-p-hydroxy-cinnamoyl-β-glucose (6), 6-O-(E )-p-hydroxy-cinnamoyl-α-glucose (7), kaempferol-3-O-β-D-rutinoside (8), and 3-O-β-D-galacopyranosyl (6-1)-α-L- rhamnosyl quercetin (9). Compounds 1 and 2 significantly reduced the intracellular content of total cholesterols and triglycerides. Conclusion Compound 1 is a new phenolic compound and exhibits potent anti-hyperlipidemic activity. Additionally, compounds 6 and 7 are isolated from D. triquetrum for the first time.

Bioanalysis and Pharmacokinetics of Eight Active Components from Huanglian Jiedu Decoction in Rat Plasma by LC-ESI-MS/MS Method

2014/8/28 9:46:22

Objective To develop a sensitive and rapid liquid chromatography?electrospray ionization?tandem mass spectrometry (LC-ESI-MS/MS) method for the simultaneous determination of wogonin, coptisine, berberine, palmatine, jatrorrhizine, phello- dendrine, magnoflorine, and wogonoside in rat plasma and to evaluate the pharmacokinetic characteristics of Huanglian Jiedu Decoction (HJD). Methods LC separation was performed on an Acquity HSS T3 column (100 mm × 2.1 mm, 1.8 μm) using gradient elution with the mobile phase consisting of acetonitrile and 0.1% formic acid water. The detection was accomplished by using positive electrospray ionization in multiple-reaction monitoring mode. Plasma samples were pretreated by protein precipitation. Results The method showed a good linearity over a wide concentration range (r2 > 0.99). The lower limits of quantification were 0.20 ng/mL for coptisine and phellodendrine, 0.48 ng/mL for berberine, 0.10 ng/mL for jatrorrhizine, 0.32 ng/mL for magnoflorine, 0.30 ng/mL for palmatine, and 4.80 ng/mL for wogonin and wogonoside, respectively. The intra- and inter-day precision of the analytes was less than 12.11%, while the accuracy was between ?14.46% and 4.86%. The mean recovery of all the analytes ranged from 93.10% to 110.91%. Conclusion This validated method offers the advantages of high sensitivity. It is successfully applied to evaluating the pharmacokinetic properties of HJD.

Quantitative Analysis of Multi-components and Volatile Contituents in Watermelon Frost Powder by HPLC and GC

Chao Zhang and Jing-hua Su — 2014/8/28 9:46:22

Objective Watermelon frost powder (WFP, Mirabilitum Praeparatum) is a Chinese patent drug commonly used for external application, consisting of dozens of Chinese materia medica. To determine the contents of the multi-components in WFP. Methods Quantitative analysis of 10 principal constituents and three principal volatile constituents had been taken in this experiment to investigate the quality of Glycyrrhizae Radix, Coptidis Rhizoma, Phellodendri Cortex, Rhei Radix et Rhizoma, Scutellariae Radix, Borneolum, and menthol in WFP by HPLC and GC. Results Comparing with raw materials, the contents of liquirtin and glycyrrhetinic acid were distinct, and the contents of rhein in WFP and raw Rhei Radix et Rhizoma were different. The possible reasons might lie in the input status of raw materials in the processes of production or others. In GC test, isoborneol, the constituent in Borneolum syntheticum, had been detected out, and should not exist in nature Borneolum. Conclusion The inspection standard of WFP should be perfected and some weakness involved in this experiment requires further explanation and research.

Rapid Analysis of Polar Components in Ophiocordyceps sinensis by Conventional Liquid Chromatography System

Wen-qing Li,Wen-jia Li,Zheng-ming Qian,Shi-lin Chen and Li Xiang — 2014/8/28 9:46:22

Objective To develop a rapid high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) method for the simultaneous determination of six polar compounds in Ophiocordyceps sinensis. Methods A poroshell SB Aq column (50 mm × 4.6 mm, 2.7 μm) and gradient elution were used; The detection wavelength of compounds was set at 260 nm. The chromatographic peaks of the six investigated compounds in sample were identified by comparing their retention times with reference compounds. Results All calibration curves showed good linearity (r > 0.999) within the tested ranges. The intra- and inter-day precisions of the six analytes were less than 0.8% and 2.1%, respectively, and the recoveries of the six analytes were between 95% and 103%. The validated method was successfully applied to the determination of six polar compounds in O. sinensis samples. Conclusion The poroshell SB Aq column is suitable for the rapid analysis of polar components in Chinese materia medica on conventional HPLC system and the developed HPLC method is also helpful to the quality control of O. sinensis.

Protective Effects of Tannins in Sanguisorbae Radix on Myelosuppression Mice

2014/8/28 9:46:22

Objective To explore the protective effects of tannins in Sanguisorba Radix (TSR) on myelosuppression mice induced by cyclophosphamide (CTX). Methods TSR was ig given at the dose of 20 mg/kg for 10 d after ip administration of CTX (200 mg/kg). Results TSR could significantly increase the numbers of white blood cells, red blood cells, and platelets of myelosuppression in mice. And it could accelerate bone marrow haemopoietic stem/progenitor cells (HSPCs) in myelosuppression mice and enhance cell proliferation by promoting cell cycles from G0/G1 phase to access into S and G2/M phases, then the reduced number of HSPCs induced by CTX was reversed. Moreover, TSR could increase the mRNA and protein expression levels of O(6)-methylguanine-DNA methyltransferase (MGMT) in HSPCs of myelosuppression mice. Conclsion TSR has a protective function against CTX-induced myelosuppression. The mechanism might be related to protecting hematopoietic stem cells of bone marrow, stimulating hematopoiesis recovery, as well as preventing the apoptosis of hematopoietic stem cells induced by CTX.

Synergistic Effects of CO2 and LED Lighting on Accumulation of Terpenes in Roots of Gynura bicolor

Jin Ren,Shuang-sheng Guo,Quan-yong Cheng,Yun-ze Shen and Hong-qi Feng — 2014/8/28 9:46:22

Objective To investigate the essential oil profile and terpenes accumulation in the roots of Gynura bicolor (Asteracese) treated by CO2 and LED lighting. Methods G. bicolor herbs were treated by CO2 at the levels of 450 (control) and 1200 (elevated) μmol/mol and LED lighting with white light, RB20 (red/blue=8/2) and RB40 (red/blue=6/4). Headspace solid-phase micro-extraction-GC MS was employed to analyze terpenes from the essential oil of the roots. Results In all treated-roots, the major components of terpenes were (E)-β-farnesene, α-, β-caryophyllene, δ-, β-, γ-elemene, and α-pinene. Increasing CO2 significantly decreased mono- and sesqui-terpenes in the roots under all light conditions, leading to the decreasing yields of the essential oils. Terpenes were at a higher level in RB20-treated roots than that in RB40-treated ones under both control and elevated CO2. Conclusion CO2 (450 μmol/mol) and 20 % blue LED lights are more conducive to the accumulation of terpenes in the roots than 1200 μmol/mol CO2 and 40% blue LED lights.

Regeneration of Carthamus tinctorius from Jimsar

Li-jiao Fan and Mei-li Guo — 2014/8/28 9:46:22

Objective To investigate the factors significantly affecting the shoot buds regeneration of Carthamus tinctorius from Jimsar (CTJ). Methods Through tissue culture experiments, the influences of culture temperature, illuminance, relative humidity, explants type, seedlings age, and plant growth regulators added in culture media on in vitro regeneration of CTJ were investigated. Results The culture temperature was set at 24 oC in daytime and 16 oC at night, illuminance at 9000 lx, and 60% relative humidity were suitable for CTJ regeneration. Cotyledons excised from 6-8 d old seedlings were more responsive than even-aged euphyll, hypocotyl, and root explants because of inducing adventitious buds. The highest percentage of regenerated shoots (79.1%) with about five adventitious buds per responding explant was obtained from MS basal medium containing 12.0 mg/L TDZ, 2.5 mg/L IBA, and 1.5 mg/L 2-ip (No. 14 medium). Regenerated shoot buds (80%) could elongate successfully after 1-2 weeks transferred to shoot elongation medium. Conclusion Suitable factors of shoot buds regeneration for CTJ were determined. It may also make a useful reference for regeneration researches of other C. tinctorius varieties in China.

Chemical Constituents in Extracts from Leaves of Lantana trifolia and Their In Vitro Anti-oxidative Activity

2014/8/28 9:46:22

Objective To isolate, purify, and analyze the anti-oxidants from the leaves of Lantana trifolia. Methods The anti-oxidative activities of the crude extracts from liquid-liquid extraction of L. trifolia leaves were assayed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) method to assess their radical scavenging and reducing abilities. The total flavonoids and phenol contents in the ethyl acetate fraction were determined by colorimetric and Folin–Ciocalteu methods, respectively. Chemical constituents were isolated from the ethyl acetate fraction and repeatedly purified using silica gel, Sephadex LH-20 column chromatography, and HPLC, respectively. The chemical structures isolated were identified by spectral analysis and chemical evidence. Results Ethyl acetate partition from liquid-liquid extraction exhibited the highest anti-oxidative activity with an IC50 value of 4.94 μg/mL, close to that of the standard (vitamin C, VC, 4.23 μg/mL). The extract was proved to contain total flavonoids and phenol contents with values of (39.0 ± 1.6) and (29.27 ± 1.46) mg/g, respectively. Six compounds were isolated and identified as kaempferol-3,7-dimethyl ether (1), verbascoside (2), apigenin (3), umuhengerin (4), ladanetin (5), and scutellarein-7-O-β-D-apiofuranoside (6). Conclusion The ethyl acetate extract from the leaves of L. trifolia possesses the potent anti-oxidative and free radical scavenging activities which are directly proportional to the concentration of phenolic contents. The anti-oxidative activity of the extract from the leaves of L. trifolia is due to its proton donating ability that converts free radicals to more stable products and terminates chain reactions. Compound 1 is isolated from the plants of Lantana Linn. for the first time. The mechanisms may be related to the therapeutic benefits of the certain traditional claims of wild L. trifolia.

Hepatoprotective Effects of Yintian Granule on Experimental Liver Injury in Mice

Yan-jun Li,Sheng Yang,Yan-jing Zhou,Fang-hong Shang and Xiao-yu Xu — 2014/8/28 9:46:22

Objective Yintian Granule (YTG), as a type of local preparation and applied for Chinese patent, is mainly composed of several traditional Chinese herbs used as both drug and food such as Lonicera macranthoides, Gardenia jasminoides, and Asparagus cochinchinenis, and has been reported to demonstrate the beneficial effects on human health in other researches. In this paper, the protective effects of YTG against experimental acute liver injury of mice were investigated to assess the value of this innovative Chinese herbal compound. Methods Carbon tetrachloride (CCl4) and 50% ethanol were used respectively to induce the acute liver injury model in mice pre- administered with YTG. Lai’s method was used to detect the level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, Coomassie brilliant blue method was used for the determination of superoxide dismutase (SOD) activity and malondialdehyde (MDA) content, and hematoxylin-eosin (HE) staining was used for the observation of liver histomorphometry. Results YTG significantly lowered the elevated ALT and AST levels, increased the SOD activity, decreased the MDA content, and inhibited the deterioration of liver. Conclusion YTG exerts protective effected against hepatocyte damage in mice induced by CCl4 and 50% ethanol, respectively.

Antihypertensive Effects of Extract from Flower Buds of Coreopsis tinctoria on Spontaneously Hypertensive Rats

2014/5/12 9:56:44

Objective To provide experimental evidence for the antihypertensive activity of the ?avonoids in flower buds of Coreopsis tinctoria (CT-F) and to investigate the underlying mechanism. Methods The spontaneously hypertensive rats (SHRs) were divided into model, captopril (positive control), and CT-F groups, and the Wistar-Kyoto rats were set as control group, eight in each group. The blood pressure of SHRs, the activity of angiotensin II (Ang-II) in plasma, nitric oxide (NO), malondialdehyde (MDA) and thoracic aorta media thickness in SHRs were measured by tail-cuff method, radioimmunity method, nitrate reductase method, thibabituric acid (TBA) method, and the hematoxylin-eosin staining method. Q-PCR analysis was performed to determine the relative quantity of angiotensin-converting enzyme (ACE), ACEII, angiotensin type 1 receptor (AT1R), and TGF-β1 mRNA in left ventricle. Results CT-F could lower the systolic blood pressure of SHRs dramatically (P < 0.01). The levels of MDA in serum and Ang II in plasma of SHRs treated with CT-F decreased markedly (P < 0.05, 0.01), the level of NO in serum increased significantly (P < 0.01). In addition, thoracic aorta media thickness in SHRs treated with CT-F was thinner than that of the model group (P < 0.05). The mRNA expression of ACE, AT1R, and TGF-β1 in left ventricle was markedly decreased (P < 0.05), while that of ACE II was increased (P < 0.05). Conclusion CT-F is effective to lower the blood pressure of SHRs, and its antihypertensive effect is probably associated with lowering the oxidative stress by reducing MDA, ameliorating aorta remodeling, dilating vessel by increasing NO and decreasing Ang-II, and regulating the expression of rennin-angiotensin System-related various genes.

Effects of Puerarin on Experimental Model of Retinal Vein Occlusion in Rats

Chun-hua Hao,Wei-ting Wang,Ming-xiang Shao,Zhuan-you Zhao and Li-da Tang — 2014/5/12 9:56:44

Objective To establish a model of retinal vein occlusion (RVO) in rats and to study the effect of puerarin on ischemic retinal disease and the corresponding mechanism. Methods RVO was induced in 10 adult male Sprague-Dawley (SD) rats by laser photothrombosis. Retinal blood flow was examined before and after 1 h of the operation, and model rats with the retinal vein blood flow decreasing by 50% compared to the basic value were chosen and then puerarins (20, 40, and 80 mg/kg) were given. The levels of vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β), and nitric oxide (NO) were analyzed. In addition, the histopathology of RVO-eyes was performed. Results RVO-eyes displayed the signs of retinal damage and ischemia on Doppler Flowmeter and histopathology. Puerarin (20, 40, and 80 mg/kg) increased blood flow by 9.3% (P < 0.05), 33.1% (P < 0.001), and 41.5% (P < 0.001)， respectively. On the other hand, the histological changes were less severe at different degrees, relieving the symptoms such as edematous and thick neuroretinal layers, lax, edematous, and disorganized optic fibers layers, swollen and confused inner and outer nuclear layer. Besides, dose-dependent decrease of VEGF and IL-1β and increase of NO in vitreous fluid were observed, with respect to the model group. Conclusion A rat model of laser photochemical-induced RVO is established and a decrease in the retinal blood flow and histological damage is detected. The puerarin has therapeutic benefit in the rat model of RVO, through the pathway of neovascularization, anti-inflammation, and increase of NO.

Anti-oxidative Activities of Ethanol Extracts from Both Wild Plant and Suspension Cell Cultures of Rheum franzenbachii

Jun-li Wang,Ming-fei Liu,Xiao-xu Li,Qian Wang,Lu Zhang and Jian-fei Li — 2014/5/12 9:56:44

Objective To evaluate the content of rhaponticin and anti-oxidative activities of the ethanol extracts from both the wild plants and suspension cell cultures of Rheum franzenbachii. Methods Quantitative analysis of rhaponticin was performed by HPLC. The anti-oxidative activities of the ethanol extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assays. Results The content of rhaponticin in the roots of the wild plant was 4.36 mg/g, while the content was only 1.59 mg/g in the leaves. The content of rhaponticin in suspension cells cultured on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 6-benzylaminopurine (6-BAP) and 2.0 mg/L 2,4-dicholorophenoxy acetic acid (2,4-D) was 17.64 mg/g, which increased by 4.05 times compared with the content in the roots of the wild plants. The roots of wild plants displayed the strongest anti-oxidative activity, followed by the suspension cells 5 and 6, and the scavenging percent was 91.96%, 91.23%, and 89.27%, respectively, at the concentration of 100 μg/mL. The IC50 values were 2.477, 15.644, and 31.415 μg/mL, respectively. In particular, the DPPH scavenging activity of the ethanol extracts from the roots of the wild plant was generally comparable to the control of ascorbic acid (VC), and the IC50 value of the extracts was lower than that of VC (2.502 μg/mL). Conclusion Rhaponticin production in the cell culture can be modulated and the accumulation can be increased. The roots of the wild plant display the strongest anti-oxidative activity. These results suggest that R. franzenbachii could hold a good potential source for human health.

Effects of Sijunzi Dripping Pill on Gastrointestinal Motility of Mice

Ying-li Wang and Yan Wang — 2014/5/12 9:56:44

Objective To study the effects of Sijunzi Dripping Pill (SDP) on gastrointestinal motility of mice. Methods The diarrhea and swimming model of mice was made by Rhei Radix et Rhizoma-induced spleen deficiency. The intestinal transit, gastric emptying test, serum motilin (MTL), vasoactive intestinal peptide (VIP), and substance P (SP) were chosen to observe the effects of high-, mid-, and low-dose SDP on stomach movements, and the water extractive of Sijunzi Decoction was used as positive control. Results Compared with the control group, the gastric emptying rate in the gastrointestinal motility group was significantly decreased, the intestinal propulsion rate was obviously increased, the levels of MTL, prostaglandin E2 (PGE2), and SP were increased (P < 0.05), while the level of VIP was decreased (P < 0.05). Compared with the model group, SDP could decrease the intestinal transit rate, whereas increase the gastric emptying rate and the level of MTL (P < 0.05); The high-dose SDP could decrease the level of PGE2 (P < 0.05) and the low-dose SDP could decrease the level of VIP (P < 0.05); Each group had no significant effect on SP. Conclusion SDP has the good effect on increasing the gastrointestinal motility of mice, and its function may partly relate to the regulation of the levels of MTL and VIP as well as PGE2.

Determination of Protopine in Rat Brain Tissues by RRLC-ESI/Q-TOF-MS Method

2014/5/12 9:56:44

Objective To analyse the quantification of protopine from Corydalis Decumbentis Rhizoma (CDR) extract in brain tissues of rats. Methods A rapid, sensitive, and accurate analytical method based on rapid resolution liquid chromatography (RRLC) coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS) was developed for the quantification of protopine in brain of rats. A simple liquid-liquid extraction process was employed for the sample preparation. Chromatographic separation was achieved using 1.8 μm porous particle columns. Results The calibration curve of protopine was linear in the range of 12–897 ng/mL. The relative standard deviations of intra- and inter-day precision values were less than 10%. The extraction recoveries were 96.4%, 99.6%, and 98.5%, for protopine at the concentration of 598.0, 119.6, and 12.0 ng/mL, respectively, and internal standard (1.27 μg/mL) was (98.60 ± 0.02)%. Conclusion The validated method is successfully applied for the determination of protopine in brain tissue of rats after ig administration of CDR extract.

Research on Hypnotic and Anticonvulsant Activities of Total Alkaloids in Leaves of Eucommia ulmoides

Xin Li,Li-peng Yang,Xin-ling Li,Li-ping Wang,Shao-yang Liu and Yan Liu — 2014/5/12 9:56:44

Objective To study the hypnotic and anticonvulsant activities of the leaves of Eucommia ulmoides (LEU), the total alkaloids were extracted from LEU using water-acid method. Methods Mice were divided into five groups, including groups contained total alkaloids in three different doses, negative and positive control groups. Direct hypnotic experiment in mice, pentobarbital sodium synergistic experiment, and anticonvulsant experiment were used. The numbers of spontaneous activities, sleep rate, sleep latency, sleep time, convulsion rate, and convulsion latency of the mice were recorded and analyzed. Results The total alkaloids from LEU could increase the sleep rate, significantly lengthen the sleep time, and shorten the sleep latency of mice. Even the low dose of total alkaloids (0.33 g/kg) showed significantly different activities with negative control group (physiological saline) with the synergistic effect of the superthreshold dose of pentobarbital sodium. Furthermore, the total alkaloids efficiently inhibited the convulsion caused by nikethamide. Conclusion The total alkaloids from LEU have the excellent sedative, hypnotic, and anticonvulsant activities in mice, with high safety and little drug side effects. Therefore, they have the protential development prospects in sedative-hypnotic drugs.

Antihyperglycemic Effect of Various Fractions from Residues of Blumea balsamifera

Yan Xia,Jian Zuo,Xiang Li and Jian-wei Chen — 2014/5/12 9:56:44

Objective To evaluate the antihyperglycemic activities of various fractions from the residues of Blumea balsamifera (BB), and to properly utilize the waste resource. Methods The antihyperglycemic activities were evaluated by the suppression on serum glucose level in vivo and α-glucosidase inhibition assay in vitro. The high-, mid-, and low-dose (1, 0.5, and 0.25 g/kg of the herb) fractions were ig given to mice for 8 d. The serum glucose was monitored at 1 and 12 h after feeding. Results The fasting and postprandial serum glucose levels of mice treated with high-dose petroleum ether fraction, ethyl acetate fraction, butyl alcohol fraction, methanol fraction, and water extract from BB were 4.45, 4.39, 4.43, 4.15, 3.74 mmol/L and 6.98, 6.23, 6.45, 6.26, 5.88 mmol/L, respectively, while those in vehicle control group were 5.63 and 7.50 mmol/L. There are four different inhibiting manners by the results of α-glucosidase inhibition assay. Conclusion The residues of BB have anti-diabetes activities after steam distillation.

Purification of Puerarin from Pueraria lobata by FCPC versus HSCCC Using Small-volume Columns

2014/5/12 9:56:44

Objective To develop an efficient method for separating and purifying puerarin from the roots of Pueraria lobata. Methods Separation by fast centrifugal partition chromatography (FCPC) was processed with a biphasic solvent system composed of ethyl acetate-n-butanol-water (2:1:3). The separation conditions were determined as follows: sample loading of 10 mg, flow rate of 2 mL/min, rotation speed of 2200 r/min, ascending mode, and detection wavelength of 254 nm. High speed countercurrent chromatography (HSCCC) was used as a comparative method with the rotation speed of 800 r/min, flow rate of 2.0 mL/min, and sample loading of 10 mg in tail-to-head mode. Results Puerarin was obtained by FCPC with a resolution of 0.90 and a purity above 99%, while a resolution below 0.50 and a purity below 90% by HSCCC. Compared with HSCCC, FCPC has the advantages with higher purity and better resolution. Conclusion FCPC is a powerful method to separate and purify puerarin.

Changes of Secondary Metabolites and Trace Elements in Gentiana macrophylla Flowers: A Potential Medicinal Plant Part

Xiao-xue Niu,Xiao-wen Chen,He Su,A. Egrinya Eneji,Yu-hai Guo and Xue-hui Dong — 2014/5/12 9:56:44

Objective To search for the potential medicinal plant part of Gentiana macrophylla based on changes of secondary metabolites and trace elements in the flowers of G. macrophylla. Methods HPLC was used to detect the changes of the active constituents (longanic acid, sweroside, gentiopicroside, and swertiamarin) and ICP-AES was used for mineral nutrients in G. macrophylla during flower development. And soluble sugar, starch, crude protein, hemicelluloses, cellulose, and lignin were determined. Results Biomass of flower in full bloom (D2) phase was considerable during flower development, in which the contents of longanic acid and gentiopicroside were at the highest levels with 2.65 and 2.88 times higher than those recorded in Chinese Pharmacopoeia 2010, sweroside and swertiamarin in the flowers were reaching 6.06 and 1.25 times higher than those in roots. Florescence is the most valuable stage during flower development. The concentration of Fe, Mg, K, P, and B was higher in the flowers than that in roots. The accumulation of active constituents in the plant was influenced by the contents of metabolically linked carbon and nitrogen compounds. Conclusion The secondary metabolites, mineral nutrients, and physicochemical indicators are tightly regulated by flower organ development, D2 is an important stage for both biomass and extraction of active constituents such as longanic acid. The flowers of G. macrophylla could be used as a potential medicinal plant part for longanic acid at a high level.

Estimation of Nuclear DNA Content in Tannin-rich Medicinal Plant Cornus officinalis by Flow Cytometry

Bo Cao,Lin-lin Zhang and Cheng-ke Bai — 2014/5/12 9:56:44

Objective The amount of nuclear DNA (C-value) is a key biodiversity character that provides strong unifying elements in revealing the phylogenetic regularity and relationship between genome size and functional traits for plant resource. The estimation of C-values could primarily extend our knowledge on the genetic background and genome diversity for medicinal plants, and thereby the variation of pharmacological constituents and phylogenetic mechanism of medicinal plant taxa will be revealed. However, a large number of medicinal plants (e.g. Cornus officinalis) typically contain a series of secondary metabolites, especially tannic acid, which would significantly affect the estimation of DNA content by flow cytometry (FCM). Methodological discussions and improvement need to be made to solve this problem. Methods Two isolation buffers LB01 and Otto 1 were selected to prepare nuclear suspension with additional treatments of pre-soaking and centrifugation combination of gradient centrifugal force and duration. The best isolation and estimation methods were determined by FCM measurement in C. officinalis. Results The dry leaves were pre-soaked in Otto I buffer for 15 min and the Otto I nuclear suspension was centrifugated at 1.0×103 g for 2 min. The results showed that debris and nuclei were better separated and the scatterplots of good quality were obtained with low coefficient of variation (CV). Contrarily, the nuclear DNA content of C. officinalis could not be accurately estimated for nuclei extracted by LB01 buffer. Finally, 2C-value and genome size of C. officinalis were first estimated as 5.92 pg and 2893 Mbp, respectively. Conclusion The new methods proposed here are able to accurately estimate DNA content of C. officinalis, which provides valuable references for the estimation of genome size in other tannin-rich medicinal plants.

Saponins from Roots of Panax notoginseng

Li-feng Han,Kaunda Joseph Sakah,Li-li Liu,Agyemang Kojo,Tao Wang and Yi Zhang — 2014/5/12 9:56:44

Objective To study the chemical constituents in the dried roots of Panax notoginseng. Methods The constituents were isolated and purified with chromatographic methods. Their structures were elucidated by spectroscopic methods (1D, 2D NMR, UV, IR, [a]D, and HRESI-TOF-MS) and chemical analyses. Results Twenty saponins including 20(S)-ginsenoside Rh1 (1), 6-O-b-D-(6′-acetyl)-glucopyranosyl-24-ene-dammar-3b, 6a,12b,20S-tetraol (2), ginseno- side Rf (3), notoginsenoside R2 (4), ginsenoside Rg2 (5), ginsenoside Rg1 (6), notoginsenoside Rt (7), koryoginsenoside R1 (8), 6-O-(b-D- glucopyranosyl)-20-O-(b-D-xylopyranosyl)-3b,6α,12b,20(S)-tetrahydroxy-dammar- 24-ene (9), pseudoginsenoside Rt3 (10), notoginsenoside R1 (11), ginsenoside Re (12), notoginsenoside N (13), ginsenoside F1 (14), ginsenoside U (15), ginsenoside Rk3 (16), 3b,12b-dihydroxydammar-(E)-20(22),24-diene-6-O-b-D-xylopyranosyl-(1→2)-b-D- glucopyranoside (17), ginsenoside Rh4 (18), pseudoginsenoside Rt5 (19), and vinaginsenoside R22 (20) were obtained. Conclusion Compounds 2, 19, and 20 are isolated from this species for the first time. The 1H-NMR data of compound 19 and 1H-NMR and 13C-NMR data of compound 20 are first reported. Meanwhile, the NMR data of b-D-xylopyranosyl group in compound 9 is corrected.

Diacylglycerol Compounds from Barks of Betula platyphylla with Inhibitory Activity against Acyltransferase

Nan Zhang,Na Li,Ya-nan Sun,Jia-lin Li,Shan-shan Xing,Zhen-dong Tuo and Long Cui — 2014/5/12 9:56:44

Objective To identify the active compounds from the barks of Betula platyphylla for inhibitory on diacylglycerol acyltransferase (DGAT1). Methods Bioassay-guided fractionation resulted in the isolation of DGAT1 inhibitory activity of lupane triterpenes. Results Ten compounds were identified as lupenone (1), lupeol (2), betulinic acid (3), betulinaldehyde (4), betulin (5), 3-deoxybetulonic acid (6), glochidonol (7), lup-20/29- ene-1β/3β-diol (8), 3α-hydroxy-lup-20(29)-en-23,28-dioic acid (9), and 3α,11α- dihydroxy-23-oxo-lup-20(29)-en-28-oic acid (10). Compounds 3-6, 9, and 10 inhibited DGAT1 with IC50 values ranging from (11.2 ± 0.3) to (38.6 ± 1.2) μmol/L. Conclusion Compounds 6, 9, and 10 are first isolated from the barks of B. platyphylla., and compounds 3-6, 9, and 10 from the barks of B. platyphylla are responsible for the inhibition on DGAT1.

A New Furan Flavonol Glycoside from Epimedium koreanum

Jin-dong Zhong,Cheng Zhou,Hong-mei Li and Rong-tao Li — 2014/5/12 9:56:45

Objective To study the chemical constituents of Epimedium koreanum. Methods Separation was carried out through silica gel and Sephadex LH-20 column chromatography and HPLC method. The chemical structures were elucidated by spectroscopic method including 1D-NMR and 2D-NMR. Results A new furanflavonol glycoside (1) was isolated and identified. Conclusion Compound 1 is a new furanflavonol glycoside, and its structure is corroborated as 5,4′-dihydroxyfurano [2″,3″:7,8] flavonol 3-O-α-L-rhamnoside.

Toxicity of Five Herbs in Aconitum L. on Tetrahymena thermophila Based on Spectrum-effect Relationship

2014/2/14 13:48:49

Objective To explore the active components with toxic effects in five Aconitum L. herbal medicines on Tetrahymena thermophila. Methods The fingerprints of five Aconitum L. herbal medicines were established by ultra-high performance liquid chromatography (UPLC) and the toxicity was evaluated by using a TAM Air Isothermal Calorimeter on Tetrahymena thermophila SB110. Results By analyzing the spectrum- effect relationships between UPLC fingerprints and toxic effects, the active components which had the toxic effects were obtained. Conclusion This work provides a general model of the combination of UPLC and microcalorimetry to study the spectrum-effect relationships of the five Aconitum L. herbal medicines, which could be used to evaluate the toxic effects and analyze the principal toxic components of the five Aconitum L. herbal medicines. On the whole, this result provides the experimental basis for the safe use of the five Aconitum L. herbal medicines in clinic.

Protective Effect of Chlorogenic Acid against Carbon Tetrachloride-induced Acute Liver Damage in Rats

Zong-xi Sun,Song Liu,Zhi-quan Zhao and Rui-qiang Su — 2014/2/14 13:48:50

Objective To evaluate the protective effect of chlorogenic acid (CGA) on carbon tetrachloride (CCl4)-induced liver injury of rats. Methods The anti-oxidative activity of CGA was investigated with several established in vitro systems. The hepatoprotective activity of CGA against CCl4-induced acute liver injury in rats was studied. The levels of alanine aminotranferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bilirubin (TB) were measured. The histopathological examination was carried out to supplement the biochemical results. Results CGA possessed strong anti-oxidative ability in vitro. The CCl4-induced liver toxicity experiment showed that the rats pretreated with CGA (300 or 500 mg/kg) had lower levels of ALT, AST, ALP, and TB than those of the CCl4-treated group. These data were supplemented with histopathological examination of rat liver sections. CGA did not show any mortality at the dose up to 5000 mg/kg. Conclusion CGA could protect the liver against CCl4-induced oxidative damage in rats, and the possible mechanism of the activity may be due to its free radical-scavenging and anti-oxidative activity.

Antidiabetic and Anti-oxidative Effects of Honokiol on Diabetic Rats Induced by High-fat Diet and Streptozotocin

Jun-jun Wang,Rong Zhao,Ji-chao Liang and Yong Chen — 2014/2/14 13:48:50

Objective To study the antidiabetic and anti-oxidative effects of honokiol (Hon) in Magnolia officinalis and its underlying molecular mechanism in diabetic rats induced by high-fat diet (HFD) and streptozotocin (STZ). Methods After ig administration with Hon [25, 50, and 100 mg/(kg·d)] to diabetic rats for consecutive 10 weeks, the levels of blood glucose (BG), oral glucose tolerance (OGT), blood lipids including total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), and low density lipoprotein-cholesterol (LDL-C), hepatic oxidative stress including the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), methane dicarboxylic aldehyde (MDA), and cytochrome P4502E1 (CYP2E1) in diabetic rats were measured. Results Compared to the diabetic control rats, ig administration of Hon resulted in significant decrease in BG, TC, TG, and LDL-C levels in serum, as well as hepatic CYP2E1 activity and MDA content in diabetic rats, whereas the level of OGT and activities of hepatic CAT, SOD, and GSH-Px in diabetic rats were significantly increased. Conclusion Hon could alleviate hyperglycemia, hyperlipemia, hepatic oxidative damage, and insulin resistance in diabetic rats by inhibiting hepatic CYP2E1 activity.

Separation of Five Quinolone Alkaloids from Fruits of Evodia rutaecarpa by High-speed Counter-current Chromatography

2014/2/14 13:48:50

Objective To develop a suitable method for the large-scale separation of quinolone alkaloids from the fruits of Evodia rutaecarpa by high-speed counter-current chromatography (HSCCC). Methods Two solvent systems were developed for the separation method. The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase at 35 oC with the flow rate of 2 mL/min and rotation speed of 855 r/min. Results Using the described method, 1-methyl-2-undecyl- 4(1H)-quinolone (1), evocarpine (2), 1-methy-2-[(6Z,9Z)]-6,9-pentade-cadienyl-4- (1H)-quinolone (3), dihydroevocarpine (4), and the mixture (5) of 1-methyl-2-[(Z)- 10-pentadecenyl]-4(1H)-quinolone (Va) and 1-methyl-2-[(Z)-6-pentadecenyl]- 4(1H)-quinolone (Vb) could be isolated from a petroleum ether extract. They were identified by 1H-NMR, 13 C-NMR, and MS/MS, and the purities were 94.3%, 95.2%, 96.8%, 98.3%, and 96.8%, respectively. Conclusion Five quinolone alkaloids from the fruits of E. rutaecarpa could be systematically isolated and purified using HSCCC. The presented method is simple and efficient with good potentials on the preparation of reference substances, especially on the quality control of Chinese materia medica.

Rapid Detection of Acetylbritannilactone from Inula britannica in Plasma of Rats by Online Sweeping-Micellar Electrokinetic Chromatography

Jing-shan Zhao,Li-xin Chen,Ya-lei Liu and Ai-ying Li — 2014/2/14 13:48:50

Objective To establish a rapid method for detecting acetylbritannilactone (ABL) by online sweeping-micellar electrokinetic chromatography (MEKC) and to elevate the sensitivity of the detection. Methods The combination of online sweeping technique with MEKC was used to determine the content of ABL in the extract of Inula britannica in plasma of rats. Results ABL was completely separated within 15 min in running buffer and sample buffer. The optimal conditions were as follows: on uncoated fused quartz silica capillary, with separation voltage of 23 kV, capillary temperature of 25 oC, and detection wavelength of 195 nm. The regression equations revealed good linear relationships between the peak area and concentration of ABL (r = 0.998), with the detection limits of 0.005–0.15 mg/mL. The relative standard deviations of migration time and peak areas for intra- and inter-batch were < 2.45% and < 2.26%, respectively. The recovery rate of this method was 96.3%–97.2%. Conclusion This method provides some advantages in separation speed, testing sensitivity, and operating convenience, with low sample and reagent consumption. The online sweeping-MEKC is an effective method for pharmacokinetic study and analysis on tracing biological samples.

Adsorption Properties for Separation of Apigenin from Viola yedoensis on LSA-10 Resin

Ai-wen Dong,Qi Zhuo,Xiao-ying Bu,Shen-hai Zhu and He Wang — 2014/2/14 13:48:50

Objective To explore the adsorption properties for the separation of apigenin from Viola yedoensis on LSA-10 resin. Methods After different types of macroporous resins were optimized, the effects of initial concentration, temperature, pH value, and other factors on resin adsorption were studied, and the kinetics and thermodynamics in the process of the static adsorption of LSA-10 resin for the apigenin separation from V. yedoensis were also investigated. Results The initial concentration of 4.0 mg/mL, temperature of 50 oC, and pH 5 were suitable for the resin adsorption, the experimental data of adsorption isotherms of LSA-10 resin were validated to fit the Freunclich and Langmuir equation, the adsorption process of apigenin was fitted to the first order adsorption kinetics equation, and the adsorption rate was mainly affected by film diffusion. The thermodynamic parameters such as adsorption enthalpy change (ΔH > 0), adsorption free energy change (ΔG < 0), and adsorption entropy change (ΔS > 0) were investigated. Conclusion The adsorption for the separation of apigenin on LSA-10 resin was an entropy-driven spontaneous process of decalescence and entropy increase, which belongs to physical adsorption. LSA-10 resin is suitable for the industrial separation of apigenin from V. yedoensis.

Identification of Icaritin Metabolites in Rats by LC-MS/MS

Lin-hu Ye,Bing-xin Xiao,Fang-rui Cao,Ying Zheng,Rui-le Pan and Qi Chang — 2015/11/25 14:14:26

Objective Icaritin is the main aglycone and also active intestinal metabolite of prenylflavonoids from the Chinese materia medica Epimedii Herba. Modern pharmacological studies have demonstrated that icaritin has a wide range of biological activities. However, its metabolites and biotransformation pathways have not yet been comprehensively investigated. The present study aims to identify icaritin metabolites in rats by using a sensitive and effective LC-MS/MS method. Methods The plasma and urine samples of rats were collected before(blank) and after oral administration of icaritin, and subjected to liquid-liquid extraction with ethyl acetate. The full-scan LC-MS chromatograms of the plasma and urine samples were compared with those of blank samples to detect the possible metabolites, which were later detected by their product ion spectra. Results A total of 23 metabolites were identified, and conjugated icaritins produced by glucuronidation, glycosylation, and sulfation were its major metabolites. Minor demethylation, hydrogenation, and oxidation metabolites were also found. Conclusion Phase II metabolism is the main metabolic pathway of icaritin.

Protection of Salidroside on Primary Astrocytes from Cell Death by Attenuating Oxidative Stress

Cun-hua Zhao,Zhi-hui Zhu,Ye-ling Wang,Georg Reiser and Li Tang — 2015/11/25 14:14:26

Objective To investigate whether salidroside(SAL) has protective and anti-oxidative effects on astrocytes. Methods Firstly, SAL was extracted from the roots of Rhodiola rosea with 70% ethanol and butanol to obtain crude phenylethyl alcohol glycosides which have been known as bioactive part of R. rosea; Secondly, WST-1 assay was carried out to assess the cell viability of astrocytes and cortical neurons under the treatment of the purified(>95%) SAL. Moreover, WST-1 assay was also used to evaluate the cytoprotective effects of SAL preventing astrocytes from staurosporine-induced cell death; Thirdly, we examined the spontaneous reactive oxygen species(ROS) and staurosporine-induced ROS generation in astrocytes in the absence or presence of SAL. Results SAL was observed to improve the astrocytes viability but not cortical neurons. In addition, SAL was able to ameliorate staurosporine-induced cell death. Moreover, SAL was able to attenuate the spontaneous ROS and staurosporine-induced ROS generation. Conclusion We here confirm that the anti-oxidative effect of SAL on primary astrocytes might be an important mechanism accounting for the cytoprotective effects from SAL.

Hypoglycemic and Hypolipidemic Effects of Lycium barbarum Polysaccharide in Diabetic Rats

Rong Zhao,Rui Jin,Yong Chen and Feng-mei Han — 2015/11/25 14:14:26

Objective To study the antidiabetic effects and the underlying molecular mechanisms of Lycium barbarum polysaccharide(LBP) and its DEAE cellulose elution fraction LBP-IV in diabetic rats induced by high fat diet(HFD) and streptozotocin(STZ). Methods After ig administration of LBP-IV[50, 100, and 200 mg/(kg·d)] and LBP[100 mg/(kg·d)] once daily for consecutive 4 weeks to diabetic rats, the glucose and lipids in blood, mRNA expression of phosphoenolpyruvate carboxykinase(PEPCK), sterol regulatory element binding-protein-1c(SREBP-1c), and fatty acid synthase(FAS) in liver were determined. Results Ig administration of LBP and LBP-IV significantly decreased the levels of blood glucose, HbA1c, TC, TG, and LDL-C, as well as the hepatic mRNA expression of PEPCK, SREBP-1c, and FAS, whereas significantly increased the oral glucose tolerance of diabetic rats. Conclusion The findings suggest that the antidiabetic effects of LBP and LBP-IV are associated with the decreased hepatic mRNA expression of PEPCK, SREBP-1c, and FAS in HFD-STZ induced diabetic rats.

Pharmacokinetics of Tetramethylpyrazine Hydrochloride in Rabbits Blood after Intranasal Administration

Jin-fan Yan,Feng Han,Li Ma,Yan-jing Cheng,Jing Gao,Jin-li Deng and Xia Feng — 2015/11/25 14:14:26

Objective To study the pharmacokinetic behavior of tetramethylpyrazine hydrochloride(TMPH) in plasma of rabbits after intranasal administration and the relationship between absorption and dosage, furthermore, to illustrate the effects of borneol and musk used in combination with TMPH on the plasma concentration profile of TMPH in rabbits. Methods The concentration of TMPH was determined by RP-HPLC method. Coumarin was used as an internal standard. Sample preparation was carried out by extraction and precipitation with methanol. The pharmacokinetic parameters were computed by software program DAS.3.1.4. Results Blood pharmacokinetics of TMPH fitted best to a non-compartment model. After intranasal administration with single dose at 10, 20, and 40 mg/kg of TMPH, the average values of C_max were 8.075, 16.537, and 33.115μg/mL, and the average values of AUC0-t were 228.93, 399.273, and 728.917 mg/(L·min), respectively. C_max of TMPH in plasma was increased by 31.136% and 38.786% compared with those without borneol and musk, and intranasal bioavailability were increased by 21.587% and 40.633% after intranasal administration of TMPH in combination with borneol, or with borneol and musk. Conclusion Borneol and musk could enhance the intranasal absorption of TMPH and increase the concentration of TMPH in blood of rabbits, especially in the early period. This work also shows the rational compatibility between borneol and musk.

Antipyretic Effects of Eupatorium chinense and Its Mechanism

Yi-ping Jiang and Jiang-ping Xu — 2015/11/25 14:14:26

Objective To investigate the antipyretic effect of Eupatorium chinense and its mechanism. Methods The content of arginine vasopressin(AVP) in ventral septal area(VSA) and blood plasma, and cyclic adenosine monophosphate(cAMP) levels of hypothalamus and blood plasma were determined by enzyme linked immunosorbent assay. Results The body temperature(Tb) was decreased at 1 h after administration of E. chinense(3 and 6 g/kg) and Aspirin(0.3 g/kg) respectively, which was significantly different from the temperature of fever model group. The antipyretic effect of Aspirin and E. chinense lasted for longer time. Aspirin(0.3 g/kg) and E. chinense(3 and 6 g/kg) reduced the level of cAMP in hypothalamus of fever rats and increased AVP content in plasma. The changes in cAMP content in plasma of all drug treatment groups were not obvious. Conclusion E. chinense has strong antipyretic effect and may affect the production of AVP and cAMP in fever rats.

Investigation of Mechanism of Premature Ovarian Failure Regulation by Kidney-tonifying Herbs and Liver-clearing Herbs in Dingjing Decoction

Xiu-feng Wang,Lei Zhang,Qing-hua Wu,Jian-xin Min,Na Ma and Lai-cheng Luo — 2015/11/25 14:14:26

Objective To investigate the mechanisms through which kidney-tonifying herbs(KTHs) and liver-clearing herbs(LCHs) in Dingjing Decoction(DJD) regulate premature ovarian failure(POF). Methods One hundred and fifty Sprague-Dawley rats were randomly divided into five groups such as control, model, KTHs, LCHs, and DJD groups. POF-related biological molecules were examined. Factor analysis was performed to investigate the regulatory networks and key biomolecules involved in mediating POF after treatment with KTHs and LCHs. Results The master regulatory factors in the reproductive endocrine network associated with KTHs intervention included four molecules in the pituitary-ovarian axis, cortisol(CORT) in the target gland of pituitary-adrenal axis, and some molecules in the hypothalamus. In contrast, the master regulatory factors associated with LCHs intervention included four molecules in the pituitary-ovarian axis and some molecules in the hypothalamus; No biomolecules in the pituitary-adrenal axis were involved in the LCH-mediated mechanisms. Gonadotropin-releasing hormone(GnRH), which was identified as a common biological molecule in the hypothalamus, was involved in regulating the reproductive endocrine network in association with KTHs intervention. Conclusion KTHs directly regulates biological molecules in the pituitary-adrenal axis and indirectly regulates those in the pituitary-adrenal axis through the hypothalamus, while the LCHs only exert its effects indirectly. GnRH is the key biological molecule associated with KTHs intervention.

Treatment of Kangxian Pills to Chronic Liver Injury in Mice Induced with Carbon Tetrachloride

Huan-tian Cui,Li Wang,Yu-hong Bian,Pei-dong Liu,Hui-fang Zhou and Zan-hong Pan — 2015/11/25 14:14:26

Objective Kangxian Pills, containing Angelicae Sinensis, Chuanxiong Rhizoma, radix paeoniae rubra, and 12 other kinds of Chinese materia medica, have the functions of softening and dispersing blood stasis. It has been used for liver injury and liver fibrosis. The current study was designed to evaluate the anti-hepatic injury activity and the mechanism of Kangxian Pills on a CCl₄-induced animal model. Methods To induce chronic liver injury, mice were treated with CCl₄ twice a week for four weeks. Kangxian Pills(6 or 12 g/kg) and Compound Biejia Ruangan Tablet(0.901 g/kg) were ig given to mice once daily for four weeks after CCl₄ was withdrawal. The anti-hepatic injury activities and mechanisms of Kangxian Pills were assessed by hepatic histology and by measuring the levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), albumin(ALB), and total protein(TP) of serum as well as superoxide dismutase(SOD) and glycogen(Gn) of the liver. Results Kangxian Pills significantly decreased the levels of liver index, ALT, and AST in mice liver injury models in treatment group. Moreover, Kangxian Pills and Compound Biejia Ruangan Tablet inhibited the CCl₄-induced reduction of SOD and Gn levels in the liver. The histological study showed that Kangxian Pills could reduce cellular swelling and infiltration of inflammatory cells in liver injury. Conclusion Kangxian Pills possess the potent abilities to alleviate chronic liver injury, suggesting that Kangxian Pills exert this effect by enhancing the anti-oxidant ability and metabolism of the liver.

Effect of Chinese Patent Medicine Naodesheng against Repeated Transient Global Cerebral Ischemia in Mice

Chun-hua Hao,Wen-gong Xi,Hai-juan Zheng,Wei-ting Wang and Zhuan-you Zhao — 2015/11/25 14:14:26

Objective To investigate the therapeutic effect and possible mechanisms of Chinese ptent medicine Naodesheng(NDS) on repeated transient global cerebral ischemia(GCI) in mice. Methods The repeated transient GCI mice were induced by bilateral carotid arteries ligation, and were randomly divided into model group, Sham group without arteries ligation, NDS groups(1.25 and 2.5 g/kg) and positive control(vinpocetine 3.1 mg/kg, VP) group. After oral administration once daily for successive 7 d, the transient GCI was induced. The degree of neurological deficit, histological changes, and neurons loss in the hippocampus were evaluated. In order to investigate the possible mechanisms, the oxidative stress and inflammatory factor were measured after 24 h of GCI. Comparison among multiple groups was performed with one-way analysis of variance(ANOVA). Results NDS could significantly alleviate the neurological function impairment, histological injury, and neurons loss, increase the superoxide dismutase(SOD) activity, decrease the content of malondialdehyde(MDA), and reduce inflammatory factor in the ischemic brain tissue. Conclusion NDS could significantly reduce brain injury induced by global ischemia, and its mechanism is closely associated with anti-oxidation and anti-inflammation.

Efficacy and Safety of Berberine in Patients with Type 2 Diabetes Mellitus:A Meta-Analysis

Xiao-chen Wei,Li-qin Zhu and Chun-ge Wang — 2015/11/25 14:14:27

Objective To assess the efficacy and safety of berberine(BBR) in patients with type 2 diabetes mellitus(T2DM) by performing a systematic review. Methods PubMed, Cochrane Library, Embase, CNKI, and CBM were searched until May 2014. The randomized controlled trials(RCTs) of the effects of BBR on blood glucose in patients with T2DM were included. The quality of RCTs was assessed by the Jadad scale, and the Review Manager 5.1 software was used for data syntheses and analyses. Results Seventeen RCTs involving 1198 patients were included. The methodological quality of these RCTs was generally low. Compared with the control groups(placebo or no intervention with medicine), BBR suggested the statistically significant benefits in improving fasting blood glucose(FBG), postprandial blood glucose(PBG), glycosylated hemoglobin, and homeostasis model assessment of insulin resistance. Subgroups analysis of BBR compared with metformin(MET) showed that 1.5g/d MET was significantly better than BBR(0.9-1.5 g/d) in lowering FBG and PBG. However, there was no significant difference between 1.5g/d BBR and 0.75g/d MET groups in blood glucose profiles. In comparison with rosiglitazone, BBR suggested the statistically significant benefits in lowering FBG. And there was no significant difference between BBR and glipizide groups in blood glucose profiles. In addition, the combination therapy of BBR and oral hypoglycemic agents had the advantages over oral hypoglycemic agents alone. No serious adverse effects of BBR have been reported. Conclusion BBR may have the beneficial effects in the control of blood glucose levels, though the efficacy of BBR is not superior to MET. BBR appeares to have advantages over rosiglitazone in improving FBG levels. In addition, the combination therapy of BBR and oral hypoglycemic agents may be a new attempt. However, the efficacy of BBR in patients with T2DM should be further evaluated by more RCTs in a larger population of patients.

Comparison on Traditional and Machinery Decoctions for Da-cheng-qi Decoction Based on Chemical Ingredients, Pharmacological Functions, and Clinical Trials

2015/11/25 14:14:27

Objective Patients and doctors often have questions about the equivalence of traditional and machinery decoctions. In this article, using Da-cheng-qi Decoction(DCQD) as a model of formula, traditional decoction(TD), machinery decoction under high pressure(MDHP), and machinery decoction under normal pressure(MDNP) were compared. Methods For chemical components, HPLC fingerprints were established and evaluated using AHP combined with CRITIC weighing method; For animals' effects, the experiments of small intestinal propulsion were conducted; For clinical effects, a randomized clinical trial(RCT) was designed and performed. Results Although there were some differences between TD and MDNP in chemical ingredients, there was no significant difference in animal experiments and clinical trials(P>0.05). Conclusion The traditional and machinery decoctions of DCQD could be used bioequivalently.

Simultaneous Determination of Four Substances in Plasma and Dermal Microdialysates of Guinea Pig after Different Acupoints Administration of Fufang Baijiezi Gel

Guo-hua Wang,Bao-xian Zhang,Qi-xia Nie,Chen Zang and Qiong Zhu — 2015/11/25 14:14:27

Objective To study the pharmacokinetic properties of Fufang Baijiezi Gel(FBG) after different acupoints administration. Methods Sinapine thiocyanate, tetrahydro-palmatine, 6-gingerol, and asarinin, which were four substances of FBG, were determined by a sensitive liquid chromatography tandem mass spectrometry method(LC-MS) both in plasma and dermal microdialysates of guinea pig simultaneously. Microdialysates were separated on an Ultimate® XB-Phenyl analytical column(150 mm×2.1 mm, 5μm) and detected by electrospray ionization(ESI) in selected ion monitoring(SIM) mode. The method was validated in terms of selectivity, linearity, sensitivity, and recovery. Result A significant difference was observed in main pharmacokinetic parameters of C_max, t_max, and AUC between acupoints administration and nonacupoints administration. Conclusion Acupoints administration resulted in a more obvious increase in bioavailability of sinapine thiocyanate, tetrahydropalmatine, 6-gingerol, and asarinin than nonacupoints administration.

Ethnopharmacological Investigation and Rapid Authentication of Mongolian Patent Medicines Digeda

2015/8/19 15:08:59

Objective To investigate Mongolian medicinal plants called Digeda and the prescriptions in Inner Mongolia region and to establish a molecular method for authentication of Digeda Mongolian patent medicines (MPMs). Methods A field investigation was conducted on traditional uses of Digeda. After interviewed traditional healers in Mongolian, ethnopharmacological information of Digeda prescriptions was recorded in detail, including names, compositions, and traditional uses. And the total DNA from 10 MPMs has been amplified by three pairs of specific primers. Specific PCR products were further identified by sequence alignment with the known sequences already submitted in GenBank or own sequences. Results Fifteen Digeda plants and 29 Digeda prescriptions with their ethnopharmacological knowledge were collected. Ten MPM samples containing Lomatogonium rotatum, Viola philippica, and Corydalis bungeana were successfully evidenced by PCR with specific bands as raw materials. Conclusion Digeda should be further investigated in ethnopharmacology, which is a fundamental step toward developing efficacious natural drugs for various diseases. PCR amplification of specific allele is an easy and economical method, which can be used to identify highly processed MPMs and will assist in monitoring their qualities and legalities.

Neuroprotection of Stilbenes from Leaves of Cajanus cajan against Oxidative Damage Induced by Corticosterone and Glutamate in Differentiated PC12 Cells

Ya-min Liu,Sheng-nan Shen,Fang-bo Xia,Qi Chang,Xin-min Liu and Rui-le Pan — 2015/8/19 15:08:59

Objective To investigate the neuroprotective effects of four stilbenes, namely cajaninstilbene acid (CSA), longistyline A, longistyline C, and cajanolactone A, isolated from the leaves of Cajanus cajan. Methods Neuroprotective effects of the four stilbenes were evaluated using rat pheochromocytoma cell line (PC12 cells) damage models induced by corticosterone (Cort) or glutamate. In order to elaborate whether the neuroprotective effects of stilbenes are related to anti-oxidant properties, both oxidant and anti-oxidant parameters were measured. Results The results of MTT assay and LDH release assay demonstrated that the four stilbenes possessed neuroprotective effects. Moreover, the treatment on PC12 cells with Cort or glutamate (Glu) could significantly increase the levels of ROS and MDA with decreasing the activities of SOD and CAT. However, the four tested stilbenes could significantly alleviate such situation by dropping out the levels of ROS and MDA, as well as enhancing the activities of SOD and CAT. Conclusion These results provide a scientific basis for further studies to explore the potential neuroprotective effects on neurodegenerative diseases.

Comparative Study for Pharmacological Action of Corydalis Rhizoma before and after Processing

Rong Li,Yan-bing Niu and Zhi-ying Dou — 2015/8/19 15:09:00

Objective To clarify the rationality and to provide reasonable usage references of processed Corydalis Rhizoma (CR) in clinic by comparing respectively pharmacological actions including analgesic, anti-inflammation, antiplatelet aggregation and spasmolysis of Yuanhu Zhitong Tablet (YZT) and Yanhusuo Decoction (YD) constituted with raw CR (RCR) or processed CR (PCR). Methods The hot-board, acetic-acid- induced twisting experiments and mouse auricular swelling model by injecting xylene in the abdomen of mouse were adopted to investigate the analgesic and anti-inflammatory effects of the four formulas on Kunming mice; The experiment of contractile activity on isolated rat intestine smooth muscle on Wistar rats was applied to observing the spasmolysis effect of the four formulas; The experiments of platelet aggregation induced by ADP or collagen on Wistar rats by turbidimetry method were used to observe the anti-platelet aggregation effect of the four formulas. Results Compared with RCR in two prescriptions, PCR displayed better analgesic effect (P < 0.05, 0.01), while there was no difference on anti-inflammatory effect; Inhibition of the two prescriptions on smooth muscle spasm was presented well, and the prescription YD with PCR was remarkable (P < 0.05, 0.01); Compared with the blank group, the two prescriptions showed outstanding effects on anti-platelet aggregation (P < 0.01), but no difference between RCR and PCR in two prescriptions. Conclusion PCR can improve the analgesic effect of prescription YZT and intensify the spasmolysis effect of prescription YD, which can be explained by the Chinese medicine processing regulation. Therefore, PCR is suggested to be chosen in YZT and YD used to cure pain symptoms.

Oleuropein Improves Long Term Potentiation at Perforant Path-dentate Gyrus Synapses in vivo

2015/8/19 15:09:00

Objective To investigate the effect of oleuropein (OE) on long term potentiation (LTP) at hippocampal perforant path-dentate gyrus synapses in vivo. Methods An outer guide cannula, a monopolar recording electrode, and a bipolar stimulating electrode were implanted in the skull and extracellular recording technique was used to record the population spike in the dentate gyrus of anesthetized rats. Results Oleuropein significantly increased the basal synaptic transmission and the amplitude of population spike was increased from (117.6 ± 2.3)% to (134.9 ± 3.7)% after administration with OE. OE also accelerated LTP induction and maintenance, the population spike amplitude after high frequency stimulation was increased from (167.2 ± 12.8)% to (225.5 ± 15.5)% and the maintenance phase of LTP was from (182.1 ± 15.1)% to (210.5 ± 9.0)% respectively after administration with OE. Conclusion Present study showed that OE significantly improved different stages of LTP, which could be the molecular mechanism of its efficacy on attenuating AD-like pathology and delaying cognitive decline. OE can be a promising drug for AD and dementia.

Protection of Sophocarpine on Colonic Barrier in DSS-induced Acute Colitis in Mice by Increasing Expression of HNF4α

2015/8/19 15:09:00

Objective To investigate the possible protective effects of sophocarpine on mucosal injury and epithelial barrier disruption on dextran sulfate sodium (DSS)-induced acute colitis. Methods Male BALB/c mice were randomly divided into three groups. The mice in normal group were given normal water, and those in model and sophocarpine- treated groups were given 2.5% DSS for 6 d to induce acute colitis. Sophocarpine (30 mg/kg) was ip administered once daily during the study period. Severity of colitis was evaluated by disease activity index (DAI), histological injury and inflammatory cytokine production including tumor necrosis factor-α (TNF-α), interleukin-1b (IL-1b), and monocyte chemoattractant protein-1 (MCP-1). The colonic barrier disruption was assessed by testing the expression of junctional adhesion molecule-1 (JAM-1), E-cadherin (E-CAD), and desmocollin-2 (DSC-2) in colon mucosa. Expression of HNF4α in colon mucosa was detected by immunohistochemistry staining and real-time RT-PCR, respectively. Results Compared with normal group, DAI, colonic shortening, and histopathological injury in model group were elevated (P < 0.05), but reduced in sophocarpine-treated group (P < 0.05). Compared with model group, the mRNA expression of inflammatory cytokines (TNF-α, IL-1β, MCP-1) were obviously lower in sophocarpine-treated group (P < 0.05), while the cellular junction proteins (E-CAD, JAM-1, and DSC-2) were higher (P < 0.05). The expression of HNF4α at mRNA and protein levels was decreased significantly in model group, but increased apparently in sophocarpine-treated group. Conclusion Sophocarpine can enhance the expression of HNF4α, promote the expression of colonic intrecellular junctions, thus, maintain the integrity of the colonic barrier and inhibit the colitis process. We suggest that sophocarpine could enhance the production of cellular junction proteins to protect the intestinal barrier fuction, at least partly, in HNF4α-dependent pathway.

Preparative Separation of Four Alkaloids from Gelsemium elegans by High-speed Counter-current Chromatography

Qiu-ping Zhang,Zheng-tao Wang and Gui-xin Chou — 2015/8/19 15:09:00

Objective To develop an efficient preparative method for the separation of Gelsemium alkaloids from Gelsemium elegans. Methods High-speed counter-current chromatography (HSCCC) with several two-phase solvent systems was investigated for the separation of Gelsemium alkaloids. The purity and structure identification of the purified compounds were performed with HPLC and NMR spectra, respectively. Results In a single operation, 206.6 mg of crude alkaloid sample was separated to yield 28.7 mg of koumine, 24.9 mg of gelsemine, 26.9 mg of humantenine, and 7.2 mg of gelsevirine, with the purities of 97.8%, 95.4%, 97.4%, and 93.5%, respectively. Conclusion A preparative HSCCC method is successfully established for the separation of four Gelsemium alkaloids from G. elegans with a modified two-phase solvent system composed of n-hexane-ethyl acetate-ethanol-0.5% triethylamine-H₂O (3:5:3:4).

C₂₁ Steroidal Saponins from Cynanchum otophyllum

2015/8/19 15:09:00

Objective To study the chemical structures of saponins in the rhizome of Cynanchum otophyllum (Asclepiadaceae), and to find new compounds. Methods The total saponins of the rhizome were separated by silica gel column chromatography. The chemical structures of obtained compounds were determined by NMR and FAB-MS spectra. Results Three C₂₁ steroidal saponins were separated. Their structures were determined as caudatin 3-O-β-d-glucopyranosyl-(1→4)-β-d-digitoxopyranosyl-(1→4)-β-d-digino- pyranosyl-(1→4)-α-d-oleandropyranoside (1), caudatin 3-O-β-d-oleandropyranosyl- (1→4)-α-d-oleandropyranosyl-(1→4)-α-d-oleandropyranoside (2), and caudatin 3-O- β-d-glucopyranosyl-(1→4)-α-d-oleandropyranosyl-(1→4)-β-d-diginopyranosyl-(1→4)- α-d-oleandropyranoside (3), respectively. Conclusion Saponins 1-3 are new compounds.

Xiyanping Injection in Treatment of Viral Pneumonia in Children: A Meta-analysis of Random Control Trials

2015/5/25 11:22:43

Objective To evaluate the efficacy and safety of Xiyanping Injection in the treatment of viral pneumonia in children. Methods We searched several databases, including PubMed, CNKI, VIP and Wanfang Data (January— June, 2014). The references of all selected studies were also retrieved to collect the relevantly randomized controlled trials (RCTs) on Xiyanping Injection for viral pneumonia in children. Two authors screened the literatures in accordance with the inclusive criteria, extracted the data and assessed the methodological quality of the included studies. We used RevMan 5.2 software for meta-analysis. Results Meta-analysis on the 10 included RCTs showed that the effective rates of defervescing and vanishing of the rashes and cough in the Xiyanping Injection group were better than those in the Ribavirin Injection group. There was no significant difference between the two groups in the incidence of adverse drug reaction. Conclusion The existing research indicated that Xiyanping Injection is a secure and efficient scheme for viral pneumonia in children. Because of the poor quality of present researches, these results should be verified by strictly-designed and large-scale sample RCTs.

Study on Integral Dissolution Model Based on Biological Potency for Compound Chinese Materia Medica

2015/5/25 11:22:42

Objective To investigate the integral dissolution model based on biological potency in order to evaluate the dissolution of Compound Chinese materia medica (CCMM) in vitro. Methods The contents of paeoniflorin, phillyrin, ginsenoside Rg1, and adenosine of ten batches of Compound Biejia Ruangan Tablet (CBRT) were determined at different times. The self-defined weighting coefficient based on the contents has been created to establish the integral dissolution model. In addition, the biological potency of CBRT was measured by MTT assay. Then, the f2 similar factor was used to evaluate the similarity of the batches. Results Compared with batch a, some batches’ f2 values of paeoniflorin and adenosine were less than 50, while f2 values of ginsenoside Rg1, phillyrin, and integral component were more than 50. Likewise, ginsenoside Rg1, phillyrin, and integral component were all in good correlation with biological dissolution. Conclusion The results of the integral dissolution based on biological test of CBRT demonstrate that the bioassay method may be a promising supplement for its quality evaluation.

Effects of Kaixin Powder on Expression of 5-HT Receptor in Hippocampus of Depressed Rats Induced by CUMS

2015/5/25 11:22:42

Objective To observe the influence of Kaixin Powder on ethology,content of 5-HT in the hippocampus, expression of mRNA, and protein in 5-HT1A and 5-HT2A receptors in the hippocampus of depressed rats induced by chronic unpredictable mild stress (CUMS). Methods Twenty-four male Wistar rats were randomly divided into blank, model, Trazodone, and Kaixin Powder groups, six rats in each group. In addition to the blank control group, other groups were established the depression model induced by CUMS combined with isolated feeding. At the same time, Trazodone group and Kaixin Powder group were treated with corresponding drugs for 3 weeks. After 3 weeks of administration, the rats were sacrificed, and a series of indexes were measured such as the contents of 5-HT, mRNA expression levels of 5-HT1A and 5-HT2A receptors, protein expression levels of 5-HT1A and 5-HT2A receptors, and so on. Results A series of indexes in the model group were decreased significantly such as the body weight growth, the sugar water intake, the score of Open Field Test, the content of 5-HT in the hippocampus, expression of mRNA, and protein in 5-HT1A receptor, while the expression of mRNA and protein in 5-HT2A receptor were increased significantly. Compared with the model group, the indexes were ameliorated in Trazodone and Kaixin Powder groups. Kaixin Powder is better than Trazodone in decreasing the level of protein in 5-HT2A receptor. Conclusion The result indicated that the depression performance of depressed rats induced by CUMS can be ameliorated by Kaixin Powder, and the mechanism maybe concerned with increasing the contents of 5-HT, exciting 5-HT1A receptor, and antagonising 5 - HT2A receptor.

Anti-inflammation of Tripterygium wilfordii Polycoride on Macrophages and Its Regulation to Inflammation via TLR4/NF-?B

2015/5/25 11:22:42

Objective To investigate the inhibitory effect of Tripterygium wilfordii polycoride (TWP) towards the pro-inflammatory factors (TNF-α and IL-1β) on the inflammatory reaction in macrophages induced by LPS and its regulatory effect and influence on the inflammation via TLR4/NF-kB. Methods The MTT method was adopted to test the effect of drugs, TWP, dexamethasone (DXM) and azathioprine (AZA) on cell growth and to select the appropriate concentration. LPS was used to induce the inflammatory reaction in RAW264.7 cell line of mice. Elisa kit was adopted to test the levels of TNF-α and IL-1β. Western blotting was adopted to test the protein expression of TNF-α and IL-1β. RT-PCR was adopted to test the expression of TLR4 and NF-?B. Results The inhibiting effect of TWP on the release of TNF-α and IL-1β in a dose dependent manner. The inhibitory effect of three different TWP dose groups is weaker than that in DXM group. However, TWP in high dose is better than AZA on TNF-α and is as strong as AZA on IL-1β. The dose dependent manner also exits in the effect on the expression of TLR4 and NF-?B, the effect is not weaker, but even stronger than that of DXM and AZA. Conclusion The research shows that down regulation of TLR4 and NF-kB p65 may be one of the mechanisms about the TWP inhibitory effect on TNF-α and IL-1β.

Total Flavonoids from Flowers of Abelmoschus manihot for Amelioration of α-naphthylisothiocyanate-induced Cholestasis by Regulating Expression of Transporters

Guo Ai1, 2,Zheng-ming Huang2 and Qing-chuan Liu2 — 2015/5/25 11:22:43

Objective To explore the molecular mechanisms of the total flavonoids extracted from the flowers of Abelmoschus manihot (TFA) against α-naphthylisothiocyanate (ANIT)-induced cholestasis. Methods The hepatoprotective activities of TFA (125, 250 and 500 mg/kg) were investigated on ANIT-induced cholestatic liver injury in rats. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), total bile acid (TBA), and bile flow were measured to evaluate the protective effect of TFA. Furthermore, the hepatic mRNA and protein levels of transports, multidrug resistance-associated protein 2 (MRP2), bile salt export pump (BSEP), and Na+-taurocholate cotransporting polypeptide (NTCP) were investigated to elucidate the protective mechanisms of TFA against ANIT-induced cholestasis. Results Pretreatment of TFA significantly and dose-dependently decreased the ANIT-induced elevation of serum ALT, AST, TBIL, and TBA levels and increased the ANIT-induced suppression of bile flow. Moreover, TFA was found to increase the expression of liver MRP2, BSEP, and NTCP in both protein and mRNA levels in ANIT-induced liver injury in rats with cholestasis. Conclusion TFA exerts a therapeutic effect on ANIT-induced liver injury in rats with cholestasis, possibly through regulating the expressions of hepatic transporters.

Inhibition of Magnolol and Honokiol on Cytochrome P450 Enzymes in Rat and Human Liver Microsomes

Jin Duan,Juan Xiao,Yong Chen and Feng-mei Han — 2015/5/25 11:22:43

Objective The purpose of this work is to evaluate the in vitro inhibitory effect of magnolol (MN) and honokiol (HN) on rat / human cytochrome P450 (CYP) enzymes (1A2/1A2, 2D/2D6, 3A/3A4, 2E1/2E1, and 2C/2C9). Methods Rat liver microsomes (RLM) and human liver microsomes (HLM) were used as the enzyme sources. After the probe substrate of each CYP isoforms was co-incubated individually with MN or HN in RLM or HLM, the metabolite production of each probe substrate in RLM and HLM incubation medium was determined and used to evaluate the activity of corresponding CYP isoforms. Results MN inhibited rat CYP1A2 and human CYP3A4 with the IC50 values of 10.0 and 56.2 μmol/L, respectively. HN inhibited rat CYP1A2 and CYP2E1, human CYP1A2 and CYP3A4 with the IC50 values of 12.1, 12.6, 17.8, and 43.9 μmol/L, respectively. Conclusion HN is a moderate or weak inhibitor of human CYP1A2. Both MN and HN are weak or non inhibitors of the other tested human CYP isoforms. The results suggest that no significant metabolic interaction seems likely to occur when the substrate drugs of CYP isoforms tested in the present work are co-administered with MN and HN.

Saponin Accumulation in Flower Buds of Panax notoginseng

2015/5/25 11:22:43

Objective Panax notoginseng is an important Chinese medicinal plant. Saponin accumulation is higher in the flower buds than in the other parts of P. notoginseng. However, the flower bud compositions have not yet been quantified. The aim of this study is to investigate the formation and accumulation of saponins in the flower buds of P. notoginseng from different populations and at different growth years. Methods Fourteen types of P. notoginseng with different growing durations and from different areas of Wenshan County, Yunnan Province were collected. We separated P. notoginseng individually into the flower buds, stems, leaves, and roots at the places where it has the highest saponin content. An efficient high-performance liquid chromatography (HPLC) method was developed for simultaneously quantifying two active saponins, ginsenoside Rb1 and ginsenoside Rb3, in the flower buds of P. notoginseng. The total saponin content was determined by using a quantitative vanillin-sulfuric acid colorimetric method. HPLC method was used to establish the fingerprints of 13 saponins and then quantify the composition in the whole plant of P. notoginseng. Results The saponin contents of different parts differ significantly, and the total saponin content and those of Rb1 and Rb3 do not entirely correlated. The flower buds of P. notoginseng contain 27.79% of total saponins, which is the highest saponin content in the whole plant. Fingerprint result showed that different saponins were appeared in different parts of the plant, i.e. flower buds, stems, leaves, and roots. Conclusion The saponin contents from the flower buds of P. notoginseng vary depending on the growth area and duration. The fingerprints show that the saponin contents and compositions vary depending on the part of P. notoginseng. These results are useful for the pharmacological evaluation and quality control of P. notoginseng.

Simultaneous Determination of Two Major Triterpenoid Saponins: Celosins ? and II in Celosiae Semen by HPLC-ELSD

Rui-bing Chen1,Yu-yang Zhang2,Jian-ming He1,Wen-dan Wu1 and Mei-li Guo1 — 2015/5/25 11:22:43

Objective To establish an analytical method for the simultaneous determination of celosin ? and celosin ??, two major triterpenoids in Celosia Semen and compare the contents of celosin ? and celosin ?? from different habitats to screen the resources of elite germplasm for further applications. Methods A sensitive and simple high- performance liquid chromatography coupled with evaporative light-scattering detector (HPLC-ELSD) method was developed for the first time for the simultaneous determination of celosin ? and celosin ??. Using this method, 21 batches of Celosiae Semen were determined from different habitats in China. Results There was an obvious difference in the contents of celosin ? and celosin ?? among Celosiae Semen species from various habitats across China. The crude drug from Yongzhou, Hunan province, Zhuhai, Guangdong province, and Nanning, Guangxi province showed the highest contents of all habitats, while Anguo, Hebei province, Haidian, Beijing, and Zhengzhou, Henan province showed the lowest content. The results also showed that geographical location had a great influence on the contents of the two compounds. The batches from lower latitudes were higher in contents of celosin ? and celosin ??. Conclusion The sun light may be a key factor influencing the contents of the two saponins, indicating that the environment has a great impact on the quality of Celosiae Semen.

A New Geranylated 2-Arylbenzofuran from Morus alba

Da-song Yang1,Zi-lei Li1,Yong-ping Yang1,Wei-lie Xiao2 and Xiao-li Li1 — 2015/5/25 11:22:43

Objective To study the chemical constituents from the root barks of Morus alba. Methods Various chromatographic techniques were used to separate the constituents and their structures were elucidated on the basis of extensive spectroscopic interpretation. Results Six compounds were isolated from the root barks of M. alba. The structure of new compound, named as albafuran D (1), was established together with known compounds sanggenon H (2), kuwanon E (3), kuwanon U (4), kuwanon F (5), and benzokuwanon E (6). Conclusion Compound 1 is a new geranylated 2-arylbenzofuran.

In Silico Molecular Docking Study of Repensine and Bentysrepinine against HBV DNA Polymerase

2015/2/26 17:16:13

Bentysrepinine (Y101), a derivative of repensine, is a novel di-peptide structure isolated from Dichondra repens. In vitro and in vivo tests exhibited that bentysrepinine markedly inhibited DNA-HBV and cccDNA activities. The binding mode of Y101 and repensine with DNA polymerase was driven by hydrophobic interactions. This might provide novel recognition of inhibitory effect of Y101 against HBV, though its inhibition mechanism needs to be validated by bio-assay at cellular level and of polymerase activity. Preliminary docking study suggested that Y101 might be able to inhibit HIV inverse transcriptase, also have the potential to interact with DNA polymerase and HCV NS5B polymerase.

Native Medicinal Plant Richness Distribution Patterns and Environmental Determinants of Xinjiang, Northwest China

2015/2/26 17:16:13

Objective To comprehensively map the distribution patterns of native medicinal plants of Xinjiang Uygur Autonomous Region, China; To find the environmental determinants and to give suggestions for the conservation planning of medicinal plants in Xinjiang region. Methods First, we compiled the distribution data of native medicinal plants at a county level in Xinjiang region, including the source plants of traditional Chinese medicine (TCM), Uygur medicine (UM) and Kazak medicine (KM); Second, we divided the distribution data into grid with a resolution of 0.1° × 0.1° and overlaid it on the topography and climate data in Arcgis 10.0; Finally, we analyzed the correlations of medicinal plant richness and environmental variables with ordinary least square (OLS) regressions and partial regressions. Results UM has more non-native species of Xinjiang than KM. The species richness of medicinal plants is high in Altay Mountains, western Tianshan Mountains and part of Kunlun Mountains. The richness of medicinal plants is highly correlated with the vascular plant species richness and climate, and further, the independent effects of vascular plant species richness are more than the independent effects of climate. The whole plant, root and rhizome, and seed and fruit are more frequently used than stem and leaf for TCM in Xinjiang region. Conclusion The distribution patterns of medicinal plants are concordant with vascular plant species, which could be carefully considered in the conservation planning of this region. Current nature reserve is a low-costing approach to conservation of medicinal plants although they were not originally established for medicinal plant protection. Nevertheless, it is urgent to further study distributions and protection status of medicinal plants in the nature reserves of Xinjiang region.

Chemical Analysis of Xueshuantong Lyophilized Powder by LC-MS Profiling

Ming Chen,Wen-zhi Yang,Wan-ying Wu and De-an Guo — 2015/2/26 17:16:14

Objective To elucidate the chemical components of Xueshuantong (XST) Lyophilized Powder and primarily disclose the chemical difference between XST and Panax notoginseng roots. Methods Liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MSn) was used to profile the saponins in XST and P. notoginseng. Structural elucidation was based on spectral analyses of negative and positive ESI-MS3 data, and the comparison of the retention behaviors. Results The optimized LC-MS profiling approach enabled resolution of major saponins. The negative mode ESI-MS3 fragmentation gave diagnostic information on the nature (neutral loss 162 Da for Glc, 146 Da for Rha, and 132 Da for a pentose) and sequence (priority: terminal > inner) of sugars and sapogenins (m/z 475 for protopanaxatriol; m/z 459 for protopanaxadiol), while the intact glycosyl portion could be characterized by characteristic Z0α+, Cnα+, and Cnβ+ (n = 2 or 3) obtained in the positive mode. Ultimately, a total of 30 saponins were characterized from XST. Compared with the roots of P. notoginseng, three malonyl-ginsenosides, ginsenoside Rd, and gyponoside XVII (or its isomer) were almost undetectable, and showed potential significance for their differentiation. Conclusion The established LC-MS profiling approach is powerful for the chemical analysis of P. notoginseng and its preparations such as XST.

Analysis on Chemical Constituents of Chinese Materia Medica Formulation Sini Decoction by HPLC-DAD-MS/MS

Liang Li and Jun-shan Yang — 2015/2/26 17:16:14

Objectives To develop a high-performance liquid chromatography (HPLC) tandem mass spectrometry method for screening and identifying the multiple constituents in the complex Chinese materia medica (CMM) formulation Sini Decoction. Methods The chemical constituents in Sini prescription as an entirety were isolated and analyzed by HPLC coupled with electrospray ionization ion-trap mass spectrometry (ESI-IT-MS) in order to clarify the chemical components of Sini Decoction and explore the chemical methods of studying CMM prescriptions. Results Twenty-five compounds were isolated from the water extract of this decoction. Through chemical and spectral analyses, the structures of 20 compounds were elucidated, including 15 flavonoids, one triterpene, two coumarins, and two other types of compounds. By using HPLC-DAD-MSn technique, the MSn cleavage behaviors of the flavonoids have been summarized as well as the origin of the isolated compounds was assigned. Conclusion The phytochemical compositon of Sini prescription is tentatively characterized by HPLC-DAD-ESI-IT-MSn. The results of the experiments demonstrate that the combination of HPLC and ESI-MSn is a sensitive and effective tool for rapidly investigating the complex CMM formulations.

Concentration Prediction of Total Flavonoids in Aurantii Fructus Extraction Process: Locally Weighted Regression versus Kinetic Model Equation Based on Fick’s Law

Yang Chen — 2015/2/26 17:16:14

Objective To predict the total flavonoids concentration of Aurantii Fructus fried with bran in its extraction process. Methods Ultraviolet spectrophotometry was used to determine the concentration of total flavonoids in different extraction time (t ) and solvent load (M ). Then the predicted procedure was carried out using the following data: 1) based on Ficks second law, the parameters of the kinetic model could be deduced and the equation was established; 2) Locally weighted regression (LWR) code was developed in the WEKA software environment to predict the concentration. And then we used both methods to predict the concentration of total flavonoids in new experiments. Results After comparing the predicted results with the experimental data, the LWR model had better accuracy and performance in the prediction. Conclusion LWR is applied to analyze the extraction process of Chinese herb for the first time, and it is totally fit for the extraction. LWR-based system is a more simple and accurate way to predict than the established equation. It is a good choice especially for a process which exists no clear rules, and can be used in the real-time control during the process.

Simultaneous Determination of Five Hydrophilic and Lipophilic Components from roots of Salvia miltiorrhiza by HPLC

Li-lan Lu — 2015/2/26 17:16:14

Objective A reversed-phase HPLC method was established for the simultaneous determination of five hydrophilic and lipophilic components in the roots of Salvia miltiorrhiza. Methods Hydrophilic components including danshensu, protocatechuic aldehyde, and salvianolic acid B, and lipophilic components such as cryptotanshinone and tanshinone IIA, were successfully separated on a Waters Symmetry C18 reverse- phase column (250 mm × 4.6 mm, 5 μm), with acetonitrile-0.5% phosphoric acid (gradient elution) as mobile phase, the detection wavelength was set at 281 nm with flow rate of 1.0 mL/min, and the column temperature was maintained at 30 °C. Results The recovery of the method was in the range of 95.1%–102.5% and the precision was less than 3% for all five analytes. All the compounds showed good linearity (R2 > 0.9990) in a relatively wide concentration range. Therefore, this HPLC method demonstrated good reproducibility, stability, and accuracy in validation studies. Conclusion Simultaneous quanti?cation of the multiple components by HPLC would be a better strategy for the quality evaluation on the roots of S. miltiorrhiza.

Antipyretic Effects of Liposoluble Fractions of Viola yedoensis

Ying-yi Pan — 2015/2/26 17:16:14

Objective To clarify the antipyretic effect of the Chinese materia medica, Violae Herba (Viola yedoensis), and its active fractions by examining the effects of V. yedoensis extracts with differing polarities on body temperature, total white blood cell (WBC) count, WBC differential count, and total serum complement of rabbits with lipopolysaccharide (LPS)-induced fever. Methods The rabbits were treated with water and ethanolic extracts of V. yedoensis, as well as petroleum ether, ethyl acetate, and n-butanol fractions of the ethanolic extract at low-, mid- and high- doses. The LPS was injected via the ear vein of rabbits in model and treatment groups 30 min post-gavage. Their body temperature was measured at 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, and 6.0 h after the LPS challenge to calculate the temperature changes and thermal response index. After the last temperature measurement, blood samples were collected to determine the blood cell counts and total serum complement (CH50) level. Results Compared with the model group, body temperature was significantly lower in the low-dose ethanolic extract group, low- and mid-dose petroleum ether fraction groups, and all three ethyl acetate fraction groups. Serum CH50 levels were lower in all treatment groups, except the ethanolic extract groups, than that in the model group, with no significant difference. V. yedoensis had no significant effect on the blood cells of febrile rabbits challenged with LPS for 6 h. Conclusion The antipyretic effects of V. yedoensis are strong, and its active fractions are the petroleum ether and ethyl acetate fractions of ethanolic extract.

Effect of Autotoxicity and Soil Microbes in Continuous Cropping Soil on Angelica sinensis Seedling Growth and Rhizosphere Soil Microbial Population

Xin-hui Zhang — 2015/2/26 17:16:14

Objective To study the effect of autotoxicity and the co-effect of autotoxicity and soil microbes from continuous cropping soil on Angelica sinensis growth, root yield and content of essential oils, and soil microbal population. Methods The pot experiments were conducted upon A. sinensis seedlings in continuous cropping soil. At the different growth stages, we determined the seedlings in growth parameters, root yield, content of essential oils, ethanol extract, and cultivable microbial populations in rhizosphere soil. Results A. sinensis seedlings were significantly inhibited in growth, root yield and quality. Compared with the control, the composition and structure of soil microbes were changed and the diversity indexes of bacteria functional groups were reduced in rhizosphere soil of A. sinensis. A. sinensis cropping problems were more seriously after the treatment with combination of autotoxicity and soil microbes than with autotoxicity alone. Conclusion The autotoxicity and soil microbes from continuous cropping soil of A. sinensis could cause the continuous cropping obstacle together.

Optimization of Supercritical Fluid Extraction and Rapid Resolution LC-MS/ESI Identification of Chromones from Saposhnikoviae Radix through Orthogonal Array Design

Zhong-ming Han,Xue Wang,Miao-miao Xu,Yun-he Wang,Li-min Yang and Mei Han — 2016/10/13 10:54:13

Objective To establish a rapid and effective supercritical fluid extraction (SFE) and rapid resolution liquid chromatography method coupled with diode-array detector (RRLC-DAD) to quantify the chromones in a species. Methods The effects of four parameters including ethanol concentration (50%-90%), pressure (25-45 MPa), temperature (40-60 oC), and time (30-90 min) on the chromones yields, namely prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, and sec-O-gluco- sylhamaudol, were investigated using SFE system with orthogonal array design (OAD). Furthermore, the extracts were analyzed using rapid resolution liquid chromatography coupled with diode-array detector (RRLC-DAD) system to confirm the results. Results Under the optimized conditions, i. e., 35 MPa of pressure, 60 °C of temperature, 70% ethanol, and 60 min of time, the yields of prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, sec-O-glucosylhamaudol, and total chromones were 3.514, 0.132, 6.242, 0.342, and 10.231 mg/g, respectively. In comparison with ultrasonic assisted extraction (UAE), SFE was able to yield a 20.7% increase in the total chromones from Saposhnikoviae Radix. Conclusion SFE is an alternative and promising method to extract chromones from this species, and the established RRLC-DAD method could serve as a rapid and effective method for the identification of chromones from Saposhnikoviae Radix.

Qualitative Analysis and Quality Evaluation of Cnidium monnieri Using UHPLC-ESI-Q-TOF/MS

Min Jia,Yi-min Li,Xin Zhai,Yang Yang and Chun-yan Li — 2016/10/13 10:54:13

Objective To identify the coumarins constituents in Cnidium monnieri and classify ten samples into three groups and this helpful chemical information could be used for the further pharmacological and clinical study on C. monnieri. Methods Qualitative analysis of coumarins in C. monnieri was detected by UHPLC-ESI-Q-TOF/MS. Quadrupole TOF/MS in either full scan mode or extracted ion mode was used for the qualitative analysis of the constituents. Relative peak area of each component was used for the hierarchical cluster analysis. Results According to UHPLC-ESI-QTOF-MS data, chemical structures of 28 coumarins in the fruits of C. monnieri were identified, including 19 simple coumarins, seven linear coumarins, and two angular coumarins. Among these constituents, ten coumarins were firstly identified in C. monnieri. In addition, hierarchical cluster analysis suggested that C. monnieri from different regions could be classified into four groups, and this clustering was correlated to the distribution significantly, Xuzhou could be regarded as the genuine producing area. Conclusion UHPLC-ESI-Q-TOF-MS is a viable method for qualitative analysis and quality evaluation of coumarins from the fruit of C. monnieri. Coumarins in C. monnieri exists intra-species variance, which indicates significant meaning for the quality control and choice of famous region drug for C. monnieri in the clinic medication.

Simultaneous Determination of Eight Active Components in Liuwei Wuling Tablet Using HPLC

2016/10/13 10:54:13

Objective To develop an effective method for simultaneous determination of eight major components in Liuwei Wuling Tablet (LWT). Methods Reversed phase HPLC method was used with variously improved conditions. Results Salidroside, forsythoside A, specnuezhenide, phillyrin, schisandrol A, schizandrol A, schizandrin A, and schizandrin B in LWT were successfully separated on a Kromasil 100-5-C18 reverse phase column (250 mm × 4.6 mm, 5 μm) using acetonitrile-0.1% phosphoric acid (gradient elution) as mobile phase. The detection wavelength was 275 nm with flow rate of 1.0 mL/min, and the column temperature was maintained at 35 oC. The recovery rate of the method was within the range of 95.13%?104.56% and the precision (RSD) was less than 3% for all eight analytes. All the compounds showed good linearities (r 2 > 0.9980) in a relatively wide concentration range. Conclusion Simultaneous quantification of the multiple components by HPLC would be a better strategy for the quality evaluation of LWT.

Simultaneous Determination of Pinoresinol Di-gluco- pyranoside and Pinoresinol Glucoside in Rat Plasma by HPLC-tandem MS/MS for Pharmacokinetic Study

2016/10/13 10:54:14

Objective Pinoresinol di-glucopyranoside (PDG) is one of the main active lignans of Eucommiae Cortex considered to be a high-quality antihypertensive drug. In this study the pharmacokinetic process of PDG and its primary in vivo metabolite pinoresinol glucoside (PG) in the portal and jugular vein were surveyed and evaluated simultaneously. Methods A sensitive high-performance liquid chromatography coupled with tandem quadruple mass spectrometry (HPLC-MS/MS) method and sample preparation protocol were developed and validated in method of selectivity, sensitivity, precision, stability, and extraction recovery for the simultaneous determination of PDG and its primary metabolite PG in rat plasma. The double intubation technique was used to simultaneously collect blood from common jugular vein and hepatic portal vein after single ig administration of PDG. Results Using this method, the quantification linearity ranges of PDG and PG in rat plasma were both 0.05-100 ng/mL. This method was successfully applied to the evaluation of the absolute oral bioavailability of PDG and determination of the pharmacokinetic properties of PDG and PG after ig administration of single dose in rats. The bioavailability of PDG at common jugular vein was 51.3% compared to that of 91.6% at hepatic portal vein. Conclusion We conclude that liver is the major conversion site of PDG to PG.

Cloning and Bioinformatic Analysis of HMGS and HMGR Genes from Panax notoginseng

2016/10/13 10:54:14

Objective To clone and analyze 3-hydroxy-3-methylglutaryl coenzyme-A synthase (HMGS) and 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR) genes from Panax notoginseng of four-year old during the flowering period, the key genes involved in the mevalonic acid pathway for saponin biosynthesis. Methods The cDNA sequences of PnHMGS1 and PnHMGR2 were obtained by reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) methods and were analyzed in their secondary structures, subcellular localizations, domains, and the three-dimensional structures of putative proteins by the bioinformatics tools. Fusion genes were constructed by the prokaryotic expression system. Results The two genes were cloned, named as PnHMGS1 and PnHMGR2, respectively, and were both predicted to be located in the chloroplast. PnHMGS1 (1410 bp) encoded a predictive unstable protein with 469 amino acids and covered hydroxymethylglutaryl-CoA synthase domain. PnHMGR2 (1690 bp) also encoded an unstable protein with 589 amino acids and possessed a hydroxymethylglutaryl-coenzyme A reductase domain and two transmembrane regions. Both of the genes were expressed most in flowers followed by roots, stems, and least in leaves. Conclusion PnHMGS1 and PnHMGR2 are firstly cloned from P. notoginseng as the new member of the HMGR family, and they show the same expression profile as P. ginseng and P. quinquefolius.

ITS2, a Better DNA Barcode than ITS in Identification of Species in Artemisia L.

Xiao-yue Wang,Si-hao Zheng,Yang Liu and Jian-ping Han — 2016/10/13 10:54:14

Objective To select a more suitable DNA barcode to identify the species in Artemisia L. Methods ITS, ITS2, and three other major universal barcode candidates (matK, rbcL, and psbA-trnH ) were evaluated in the identification efficiency using a total of 1433 sequences downloaded from GenBank representing 343 species in Artemisia L. ITS and ITS2 were evaluated in the PCR and sequencing rate, sequencing peak quality (Q value), and misread rate. One hundred and twelve A. annua samples were collected from 11 populations across over China, which were amplified with universal primers on the ITS and ITS2 regions. Results ITS and ITS2 shared a higher identification efficiency and exhibited 71.43% and 64.11% detectability at the species level, respectively. The Q values of ITS and ITS2 showed that the direct PCR sequencing data were reliable for the ITS2 region and ITS exhibited poor sequencing trace quality. In certain sites, the ITS sequences exhibited reading ambiguities and errors, indicating that the misread and deletion sites in the ITS region would incorrectly inflate the identification ratio. Conclusion ITS2 is a suitable barcode for identification of species in Artemisia L., which enlarges the optimal range of divergence levels for taxonomic inferences using ITS2 sequences.

Effects of Berberine on Hepatic Sirtuin 1-uncoupling Protein 2 Pathway in Non-alcoholic Fatty Liver Disease Rats Induced by High-fat Diet

Yuan-jun Deng1,Yu-pei Zhang1,Qin-he Yang1,Li Han2 and Yin-ji Liang1 — 2016/10/24 15:31:17

Objective To investigate the involvement of sirtuin 1 (SIRT1)-uncoupling protein 2 (UCP2) pathway in the development of non-alcoholic fatty liver disease and whether berberine exerts its effects by regulating this pathway. Methods Male SD rats were divided into three groups: normal control group, high-fat diet group, and berberine supplement group. The rats in the normal control group were given normal diet while the rats in the other two groups were fed with high-fat diet. Rats in the berberine supplement group were concurrently given berberine (100 mg/kg body weight) once daily. After 16 weeks, the levels of serum, liver lipids, and serum aminotransferase were measured using an automatic biochemical analyzer. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the liver were measured using commercial kits. Histopathological changes of liver tissues were observed by hematoxylin and eosin (HE) staining and Oil Red O staining. The hepatic mRNA and protein levels of SIRT1 and UCP2 were assayed by reverse transcription polymerase chain reaction (RT-PCR) or Western blotting. Results Berberine supplement could significantly decrease the serum and liver lipid contents in rats fed with high-fat diet. Meanwhile, SOD level was significantly elevated, but MDA level was reduced in the liver. The results of HE and Oil Red O staining showed that the hepatic steatosis was alleviated in berberine supplement group. Furthermore, berberine induced an increase in SIRT1 expression but a decrease in UCP2 expression. Conclusion The regulation of hepatic SIRT1-UCP2 pathway may be an important mechanism by which berberine exerts the beneficial effects in NAFLD rats.

1,5-O-Dicaffeoyl-quinic Acid as a Novel Potential NMDA Receptor Inhibitor from Traditional Chinese Medicine Database by Virtual Screening

2016/10/24 15:31:17

Objective Neurodegenerative diseases, such as ischemia, traumatic injury, Alzheimer’s disease, and Parkinson’s disease are characterized by neuronal loss and dysfunction. It is known that glutamate-induced toxicity plays an important role in neurodegenerative diseases. Glutamate toxicity seems to be mediated by excessive influx of Ca2+ into neuronal cells through activation of N-methyl-D-aspartate (NMDA) receptor. To search for potential NMDA receptor inhibitors in traditional Chinese medicine. Methods A series of computer methods including drug-likeness evaluation, ADMET tests as well as molecular docking have been used. Results 1,5-O-dicaffeoyl-quinic acid was identified as NMDA receptor inhibitor by virtual screening. Its neuroprotective activity was further confirmed by in vitro test. 1,5-O-dicaffeoyl-quinic acid showed strong neuroprotection against NMDA-induced cell injury. Conclusion 1,5-O-Dicaffeoyl- quinic acid may be regarded as a potential NMDA receptor inhibitor for the prevention and treatment of neurodegenerative disorders.

Preparation of Rutin-liposome Drug Delivery Systems and Evaluation on Their in vitro Antioxidant Activity

2016/10/24 15:31:17

Objective To prepare, characterize and evaluate the antioxidant activity of rutin-liposome (RL). Methods Liposomes of rutin were prepared by film dispersion method and the encapsulation efficiency (EE) was determined by RP-HPLC. Human umbilical vein endothelial cells (HUVECs) were injuried by H2O2 and treated with either free aqueous rutin or the RL delivery systems. The viability of HUVECs was determined by MTT and ELISA. Results The drug delivery system showed uniform rutin loaded nanoparticles with average particle size of (147.20 ± 1.42) nm, polydispersity index of (0.191 ± 0.003) nm, Zeta potential of (-20.0 ± 1.0) mV, and the drug EE was closed to 90.0%. The antioxidant effect of the drug delivery system to H2O2-damaged HUVECs showed that RL could increase injury cells viability compared to free aqueous rutin, which was accompanied with an obvious decrease in malondialdehyde (MDA), lactate dehydrogenase (LDH) while increase the level of nitrogen oxide (NOS). Conclusion The nanostructured RL is improved on the antioxidant effect and may be treating the different diseases caused by free radicals.

Effects of Coptidis Rhizoma on Cell Cycle, DNA Damage, and Apoptosis in L929 Murine Fibroblast Cells

Yan-fei Huang,Man-man Gu,Jing Xu,Chun-yang Han,Teng-fei Liu and Cui-yan Liu — 2016/7/21 17:23:33

Objective Coptidis Rhizoma (CR), a widely used traditional Chinese herbal medicine, is commonly believed to be non-toxic. However, little is known about its cytotoxicity and relevant mechanisms at cellular and genetic levels. The present study was conducted to explore the cytotoxicity of CR and its mechanisms related to cell cycle arrest, DNA damage, cell apoptosis, and mitochondrial membrane potential in L929 murine fibroblast cells. Methods The cells were cultured and treated with different concentration of CR aqueous extract for 24 h. Cell viability was determined by CCK-8 method, morphological changes, and mitochondrial membrane potential were observed with an inverted microscope, cell cycle and cell apoptosis were examined by flow cytometry and DNA damages were detected by comet assay. Results Our results showed that cell viability was significantly decreased in a dose-dependent manner when concentration was higher than 0.2 mg/mL. A concentration above 1 mg/mL altered the cells morphology. Each DNA damage indicator score increased in the groups with the concentration of above 0.1 mg/mL. Cells at G2/M phase, cell apoptosis and mitochondrial membrane potential changed in the 2 mg/mL group. Conclusion Overall, our study suggests that CR at a high dosage exhibits cytotoxicity on L929 cells, which is likely to be the consequences of cell cycle arrest, DNA damage, cell apoptosis and mitochondrial membrane potential reduction.

Effects of Zhengtian Pills on Migraine Headache in Rats via Transient Receptor Potential Vanilloid 1

2016/7/21 17:23:33

Effect of Polygoni Cuspidati Rhizoma et Radix and Its Ingredient Resveratrol on Experimental Autoimmune Myasthenia Gravis by Suppressing Immune Response

Shen Liu1,Xin-xin Zhang2,Shan Zhuang3,Chun-hong Li4 and Yan-bin Li5 — 2016/7/21 17:23:33

Objective To investigate the effects of Polygoni Cuspidati Rhizoma et Radix (PCRR) and its ingredient resveratrol (Res) on experimental autoimmune myasthenia gravis (EAMG). Methods EAMG was induced in Lewis rats by the immunization of a synthetic peptide corresponding to region 97–116 of the rat acetylcholine receptor (AChR) α subunit (R97-116). EAMG rats were randomly divided into PCRR group, Res group, and control (C) group, and were ig administered respectively with PCRR (2 g/kg), Res (20 mg/kg), and DMSO (0.4 mL/kg) every day from day 5 after immunization to day 42. Clinical evaluation, lymphocyte proliferation, cytokines, and anti-97-116 antibodies were performed for examination of their therapeutic effects. Results Treatments with PCRR and Res significantly ameliorated clinical symptoms, down-regulated TNF-α and up-regulated IL-10 in serum and culture supernatants of lymphocytes stimulated with R97-116, and decreased levels of anti-R97-116 IgG1 and IgG2a in serum compared with C group. Unexpectedly, PCRR but not Res inhibited lymphocyte proliferation compared with C group. Conclusion PCRR and Res ameliorating EAMG is associated with suppressing immune response, and indicates a therapeutic potential for EAMG and even human myasthenia gravis (MG). Res may be the main effective ingredient from PCRR ameliorating EAMG, but further experiments are necessary.

Inhibition of Ovalitenin A on Proliferation of HeLa Cells via Apoptosis, G2/M Cell Cycle Arrest, and Down- regulation of COX-2

Dong-yu Liu1,Yan Guo1, 2,Jian-yong Si1,Gui-bo Sun1,Bing Zhang2 and Li Cao1 — 2016/7/21 17:23:33

Objective Ovalitenin A (1-(4-methoxybenzofuran-5-yl)-3-phenyl-2-propen-1-one) is a chalcone isolated from Millettia pulchra. The aim of the study was to investigate the antitumor effect of ovalitenin A on apoptosis in vitro and in vivo and to identify the mechanism involved. Methods The effect of ovalitenin A in human cervical cancer HeLa cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, morphological observation, flow cytometric measurement, Western blotting, and xenograft model. Results Ovalitenin A inhibited the proliferation of HeLa cells in a dose-dependent manner in vitro and in vivo and induced the apoptosis evidenced by characteristic apoptotic morphological changes, phosphatidylserine externalization, and activation of caspase-3. In addition, ovalitenin A induced G2/M cell cycle arrest and up-regulation of the Bax/Bcl-2 ratio. Furthermore, ovalitenin A decreased protein level of COX-2 and induced the loss of mitochondrial membrane potential. Conclusion These data suggest that ovalitenin A has the potential of anticancer properties for the treatment of cervical cancer.

Effects of Zhengtian Pills on Migraine Headache in Rats via Transient Receptor Potential Vanilloid 1

2016/7/21 17:23:33

Objective To investigate the effects and molecular mechanism of Zhengtian Pills (ZTP) on migraine headache. Methods All rats were randomly divided into control, positive control, migraine model, low- and high-dose ZTP groups, and glyceryl trinitrate was injected to induce migraine headache. The time of ears turning red, frequency of scratching head, climbing the cage, and head-twitching were used to evaluate rat behaviors. After 10 d administration of ZTP, the expression levels of transient receptor potential vanilloid 1 (TRPV1) both in cortex and hippocampus were determined by Western blotting. Results After 2 min of glyceryl trinitrate injection, rats showed headache phenomena that parallels the clinical symptoms of migraine, which peaked in 30 min, and lasted for 60 min. Frequency of head-twitching and numbers of scratching head in glyceryl trinitrate (GTN) group were significantly increased. In contrast, after ZTP (1.08 g/kg, ig) treatment, the numbers of scratching head with fore-limb, hind-limb and the frequency of head-twitching were significantly decreased. Flunarizine (FLU) and low-dose ZTP (0.54 g/kg) also showed a trend to decrease the numbers of scratching head and head-twitching frequency, but no significant difference. Besides, ZTP significantly decreased the up-regulated TRPV1 protein expression level both in cortex and hippocampus. Conclusion The present study shows that ZTP could significantly improve the migraine symptoms of headache in rats and TRPV1 might be one of the important molecular mechanisms. This is the first report about the effect of ZTP on TRPV1 protein expression level both in cortex and hippocampus of rats.

Evaluation of Antinociceptive Activity of Methanol Extract from Cleome rutidosperma in Mice

2016/7/21 17:23:33

Objective Cleome rutidosperma (Capparidaceae), commonly known as “Fringed Spider Flower”, is a medicinal plant found in Southeast Asia. C. rutidosperma is used in folk medicine for diuretic, laxative, analgesic, anti-inflammatory, antipyretic, antimicrobial, anti-oxidant, hypoglycemic, and anthelmintic activities. We have evaluated the anti-nociceptive properties of methanol extract from C. rutidosperma (MECR) in vivo. Methods Thermal method (hot plate test and tail flick test) was induced to judge the anti-inflammatory effect and couple of chemical method also used (formalin induced licking test; writhing test carried by acetic acid) to evaluate analgesic effect. Both of these tests were made over animal models, like mice and rats. Two different doses (100 and 200 mg/kg) were used for each case of test, while morphine sulphate (5mg/kg, ip) was used as reference drug. Results MECR demonstrated the significantly anti-nociceptive activity in the analgesic and anti-inflammatory tests by reducing nociception in mice models (P < 0.001). In the hot-plate and tail-flick tests, MECR significantly elongated the time to response to the thermal stimuli (100 and 200 mg/kg with P < 0.05, 0.001). The remarkable increase in the latency was observed at 90 and 120 min. In acetic acid-induced writhing test and formalin induced licking test for anti-inflammatory activity, MECR at 100 and 200 mg/kg doses exhibited significant (P < 0.001) reduction of writhing and licking response. Conclusion The anti-inflammatory and analgesic effects of C. rutidosperma propose that this effect may be a result of both peripheral and central mechanisms. Further study is required to ensure the proper mechanism of action as well as the active ingredient.

Simultaneous Determination of Eight Constituents in Fruits of Rubus chingii by UPLC

2016/7/21 17:23:33

Objective To develop a simple, efficient, and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii, which is widely used in Chinese materia medica (CMM), known as Fupenzi (FPZ) in Chinese. Methods An ultra performance liquid chromatography-photo diode array (UPLC-PDA) system was employed for simultaneous quantification of eight compounds, i. e. adenosine, gallic acid, brevifolin carboxylic acid, ethyl gallate, ellagic acid, kaempferol-3-O-rutinoside, kaempferol-3-O-b-D-glucopyranoside, and tiliroside. The chromatographic analysis was performed on a C18 column using a gradient elution of acetonitrile -0.1% formic acid aqueous solution within a runtime of 25 min. Results All calibration curves were linear (R2 > 0.9997) over the tested ranges. The intra- and inter-day precisions as determined from sample solutions were both less than 2.45% and 2.78%, respectively. The average recoveries for the eight constituents ranged from 94.77% to 101.35% with RSD ￡ 4.41%. The newly-developed method was applied to the quality assessment of various R. chingii samples, including both ripe and unripe fruits of R. chingii from different habitats. Conclusion The relative levels of the investigated compounds vary remarkably in the fruits of R. chingii collected from different habitats. As only two of the eight compounds, adenosine and ellagic acid, are determined in the ripe fruits of R. chingii, the results may explain the reason why only the unripe fruits can be used in CMM.

Comprehensive Evaluation of Powdered Chinese Herbal Medicines-An Exemplification of Isatidis Radix

2016/7/21 17:23:33

Objective Currently, powdered Chinese herbal medicines (CHMs) were mainly evaluated through physical property, chemical dissolution, and bioactivity independently. It could not reflect the quality comprehensively. This paper was to explore and establish a comprehensive evaluation method for powdered CHMs. Methods Isatidis Radix was chosen as an exemple. Firstly, powdered Isatidis Radix in different particle size was prepared. Then, their physical properties were characterized. The dissolution of index component epigoitrin was determined, and their antiviral activities were evaluated by neuraminidase-based bioassay. Results As the particle size decreased, powder distribution tended to be uniform, and the dissolution of epigoitrin increased, antiviral activity enhanced. According to cluster analysis of above results, the sequence of evaluation consequence was ultrafine powder S2 (D90: 32.80 ± 0.29) > ultrafine powder S1 (D90: 52.08 ± 0.53) > fine powder S0 (D90: 118.16 ± 0.76) (from the superior to the inferior). Conclusion Overall, the comprehensive evaluation for powdered CHMs based on the physical characterization, chemical dissolution, and bioassay could not only be used to evaluate powdered herbs, but also guide the screening and optimization of the particle size of powder.

Screening and Validation of Active Ingredients in Sini Decoction by Combination Method of Pharmacophore Modeling and Molecular Docking

Lang-dong Chen,Dong-yao Wang,Yan Cao,Zhen-yu Zhu and Yi-feng Chai — 2016/4/28 14:29:36

Objective To screen the active compounds in Sini Decoction showing the potential to inhibit tumor necrosis factor α (TNFα) to alleviate Doxorubicin (DOX)-induced heart failure. Methods A chemical database of Sini Decoction was constructed from literature research. The generated pharmacophore models based on TNF-α used to screen active ingredients of Sini Decoction in the database by Discovery Studio 2.5. Molecular docking by Autodock 4.2 was adopted to demonstrate the hit compounds’ affinities with TNFα. Furthermore, DOX-induced heart failure model on H9c2 cell line was constructed and cell viability was detected by CCK-8 to validate the therapeutic effect of potential active compounds. Results The higenamine showed potential cardiovascular protective effect through virtual screening. And the activity was identified in vitro. Conclusion In this study, we found that higenamine may inhibit TNF-α through virtual docking and validated that higenamine may have the potential of treatment for heart failure in the model of doxorubicin-induced myocardial toxicity to H9c2 cells.

Investigation on Endogenous Metabolites in Pancreas of Diabetic Rats after Treatment by Genipin through 1H-NMR-based Metabolomic Profiles

Ting Ling-hu1, 2,Xiao-li Shen1, 2,Jun-sheng Tian1 and Xue-mei Qin1 — 2016/4/28 14:29:36

Objective To investigate the change rules of endogenous metabolites in pancreas of the diabetic rats, and to explore the mechanism of genipin treatment for diabetes rats. Methods Metabolomic method based on 1H-NMR was applied, the diabetic rat model was prepared by ip injecting alloxan, and the high-, mid-, and low-dose genipin or metformin hydrochloride was ig injected as well as the rats in control and model groups were given the same volume of normal saline for 2 weeks. The pancreases of rats were collected and 1H-NMR test was conducted, the metabolomic technology was adopted to analyze the endogenous metabolite changes in pancreas. Results The high-dose genipin possessed a better hypoglycemic effect, which could increase the contents of isoleucine, phosphatidylcholine, and phosphatidylethanolamine, and significantly reduce the contents of lactic acid, alanine, glutamine, aspartic acid, and creatine in pancreas of diabetic rats. Conclusion This study provided a theoretical basis for further exploration on the pathogenesis of diabetes and the mechanism of genipin for treatment of diabetes.

Molecular Simulation Study on Bentysrepinine Metabolites Improving Binding Affinity of HBV DNA Polymerase

2016/4/28 14:29:36

Objective To study the effect of bentysrepinine (Y101) metabolites on improving binding affinity of HBV DNA polymerase. Methods The binding mode of Y101 and its metabolites with DNA polymerase has been driven by hydrophobic interaction. Results Two compounds, T2 and T4, exhibited the improvement of the binding affinity to HBV DNA polymerase protein, which suggests that the inhibitory activity against HBV DNA polymerase protein can be enhanced. Conclusion The variant docking poses of T2 and T4 might imply the novel recognition of inhibitory effects of T2 and T4, in comparison with Y101.

Improving Effects of Astragalus Polysaccharides on Cardiac Function via Keap1/Nrf2-ARE Signal Pathway in Adjuvant Arthritis Rats

Yue Sun1,Jian Liu2,Lei Wan2,Fang Wang1,Xiao-jun Zhang1 and Ya-jun Qi1 — 2016/4/28 14:29:36

Objective To observe the effect of Astragalus polysaccharides (APS) on cardiac function and expression of Keap1/Nrf2-ARE signal pathway in adjuvant arthritis (AA) rats. Methods Forty-eight rats were randomly divided into normal control (NC) group, model control (MC) group, leflunomide (LEF) group, and APS group. The AA model rats were induced with Freund’s complete adjuvant, then administration began from day 19 after modeling for 30 d. The paw swelling and arthritic index (AI), the cardiac function indexes, the expression of Keap1, maf, Nrf2 mRNAs, and HO-1, r-GCS proteins in cardiac tissues were observed. Oxidation-related substances (SOD, MDA, ROS, and TAC) and cytokines (IL-10 and TNF-α) were also determined. Results Compared with NC group, the paw swelling and AI were increased, and the body weight was decreased in MC group. HR, HI, LVSP, LVEDP, and levels of MDA, TAC, ROS, RNS, and TNF-α in MC group were increased, while dp/dtmax and levels of GSH, TRX, and IL-10 were decreased. Compared with MC group, LEF group showed higher HR and ± dp/dtmax, and lower LVEDP. In APS group, the HR, LVSP, and LVEDP were decreased and ±dp/dtmax was increased. TNF-α was decreased, TAC, ROS, MDA, and IL-10 were increased in both LEF and APS groups. APS and LEF groups also showed less MDA, TAC, ROS, and SOD, and the differences were statistically significant. Spearman correlation analysis showed that the cardiac function parameters were positively correlated with Keap1, anti-oxidant indicators, and anti-inflammation cytokines, and negatively correlated with Nrf2, oxidation indictors, pro-inflammation cytokines, and inflammation indictors. Conclusion APS can adjust the expression of Keap1/Nrf2-ARE signal pathway in AA rats, and can also improve their cardiac function. The mechanism may be involved in increasing myocardial anti-oxidant capacity, reducing oxidative stress, and inhibiting inflammation.

Rapid Analysis and Identification of Absorbed Components and Their Metabolites of Yuanhu Zhitong Dropping Pill in Rat Plasma and Brain Tissue Using UPLC-Q-TOF/MS with Multivariate Statistical Analysis

2016/4/28 14:29:36

Objective To establish an effective approach for rapid and comprehensive analysis on the absorbed and metabolic components in rats after ig administration of Yuanhu Zhitong Dropping Pill (YHZT). Methods Based on the combination of UPLC-Q-TOF/MS and multivariate statistical analysis, the absorbed prototype constituents and their metabolites in rat plasma were rapidly analyzed and identified, and the components absorbed into brain were further identified by comparing the extracted ion chromatograms (EICs) of control and brain tissue samples of dosed rats. Results A total of 38 YHZT-related xenobiotic compounds were detected and identified as the potential bioactive constituents in rat plasma, including 24 absorbed prototype constituents and 14 metabolites. In particular, of all prototype constituents, 14 were also detected in rat brain tissue, indicating that they could penetrate the blood-brain barrier and enter into brain. Conclusion An effective method is established and applied to analyze the potential bioactive constituents in YHZT, which provides a pathway to further investigate the pharmacological pattern and mechanism of YHZT.

Induction of Solasonine on Apoptosis of Human Breast Cancer Bcap-37 Cells through Mitochondria-Mediated Pathway

Na Li1, 2,Liang Cao1, 2,Yan-ru Wang2,Xiao-qian Tao2 and Zhen-zhong Wang1, 2 — 2016/4/28 14:29:37

Objective To study the in vitro antiproliferative effect and probable mechanism of solasonine on human breast cancer Bcap-37 cells, meanwhile, make comparison with solamargine. Methods The cytotoxicity was evaluated by MTT assay. The cell damage and type of cell death were examined through Hoechst33342/PI and Annexin V/PI staining, respectively. Mitochondrial membrane potential was detected by JC-1 staining. The expression of Bcl-2, Bcl-xL, Bax, and cytochrome c was determined by immunoblot method, and the activation of caspase-3 was analyzed by immunocytochemistry method. Results Solasonine showed the different extents of cytotoxicity on eight human tumor cell lines as well as four human normal cell lines, and the IC50 values of solasonine ranged from 12.73 to 37.15 μmol/L. Cell apoptosis and mitochondria depolarization were observed in Bcap-37 cells after treatment with solasonine for 24 h, respectively. In immunoblot and immunocytochemistry analysis, solasonine obviously induced the up-regulation of Bax and down-regulation of Bcl-2 and Bcl-xL, caused the release of cytochrome c from mitochondria into cytosol, and increased the expression of both pro- and cleaved caspase-3. Solamargine exhibited stronger antipoliferative activity than solasonine, but the similar mechanism in Bcap-37 cells in this study. Conclusion Solasonine possesses the antiproliferative effect on tumor cells. Regulation of the levels of Bcl-2, Bcl-xL, Bax, and activation of mitochondria cytochrome c-dependent apoptosis pathway might be one of its main antitumor mechanisms against breast cancer cells. In view of the cytotoxic effect of solasonine and solamargine also shown on normal cells, the safety needs concern when the antitumor activity is studied.

Effect of Chinese Materia Medica with Tonifying Kidney Function on Transplantation of Multipotency Mesenchymal Stem Cells from Human Umbilical Cord in Mice Model of Acute Kidney Injury

Xiao-qian Chu1,Li Wang2,Wen Li1,Paulina Duya3 and Yu-hong Bian1 — 2016/4/28 14:29:37

Objective Mesenchymal stem cells (MSCs) represent a promising population for supporting new clinical concepts in cellular therapy, and they can be reprogrammed into induced pluripotent stem cells (iPSCs) by defined factors. Methods This method opened up a new era of stem cell research, because the transplantation rejection of iPSCs is the bottleneck of its clinical application, so seeking alternative compounds and animal origin diagnostic reagents to achieve full chemical iPSCs is to be done to solve this problem. Results The application of these iPSCs has largely been associated with well known undesirable effects such as the development of cancers in certain experimental models. This has called for the search and use of reprogramming factors that are safe. Chinese materia medica (CMM) with tonifying kidney function (TKF) offers an alternative source. On the other hand, human umbilical cord mesenchymal stem cells (hUMSCs) are known to be a “young” source of MSCs, hUMSCs transplantation is an attractive approach for acute kidney injury repair. Therefore, In this study, we investigated whether the treatment of CMM with TKF on hUMSCs could enhance the repair in mice model of acute kidney injury after transplantation. Conclusion Our results showed that the treatment of hUMSCs with kidney tonifying CMM increased their multipotency, improved the renal function of mice and enhanced subsequent homing to the injured kidney in an acute kidney injury mice model.

Enzyme-assisted Extraction and Enrichment of Galanthamine from Lycoris aurea

2016/4/28 14:29:37

Objective To explore the optimum condition for complex enzyme-assisted extraction of galanthamine from Lycoris aurea by L9 (34) orthogonal array design and separation effect of cation exchange resin on galanthamine. Methods Cellulase and pectinase solution was used as the extraction solvent. The effects of pH value of enzyme, amount of complex enzyme, dissociation time, and enzymatic hydrolysis temperature on the extraction results were investigated. Results The optimal conditions were obtained as follows: ratio of solid to liquid (g: mL) 1:10, pH value 4.5, amount of complex enzymes 4%, enzymatic hydrolysis temperature 50 oC, and reaction time 2.0 h. Under these conditions, the extraction yield of galanthamine was 0.0294%. In addition, D-001 cation exchange resin was selected for separation of galanthamine. The separation conditions were that adsorption flow rate was 3 BV/h with reagent of pH 2 and the desorption flow rate was 3 BV/h with 70% ethanol solution containing 1.5 mol/L ammonia. After separation, the content of galanthamine was increased to 12.31%. Conclusion The results provide a reference for industrial production of galanthamine.

Cell Cycle Arrest, Apoptosis, and Autophagy Induced by Chabamide in Human Leukemia Cells

Kun Hu,Meng Yang,Yuan-yuan Xu,Kun Wei and Jie Ren — 2016/1/28 16:54:55

Objective To investigate the antitumor effect of chabamide in K562 (human leukemia cell line) cells. Methods The cytotoxicity was assessed by a standard colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The morphological changes were observed by Hoechst 33258 staining. Induction of apoptosis, loss of the mitochondrial membrane potential (ΔΨm), and cell cycle analysis were evaluated by flow cytometry (FCM) analysis. Levels of apoptosis-related proteins, cell cycle-related proteins, and LC3 protein were detected by Western blotting. Moreover, the autophagy induced by chabamide was also detected by MDC fluorescent staining. Results Chabamide significantly inhibited cell proliferation by cell cycle arrest in the G0/G1 phase. This phenomenon was associated with an obvious increase in p21 expression and decrease in cyclin D1 and cyclin-dependent kinase 2/4/6 protein expression. Moreover, chabamide could regulate the changes in mitochondrial membrane potential, increase the expression of apoptosis-related proteins, such as Bax and cytochrome C, and decrease the protein expression of Bcl-2, caspase-9, caspase-3, and PARP-1. JNK, ERK1/2, and p38 were also regulated by chabamide in K562 cells. Furthermore, induction of autophagy, marked by autophagic vacuole formation, was detected. Conversion of LC3-I to LC3-II, a marker of active autophagosome formation, was also detected following chabamide treatment. Conclusion The antitumor activity of chabamide with the results of apoptosis and autophagy induction was confirmed in K562 cells.

PERK Signaling of Unfolded Protein Response Activated in Acute Hypobaric Hypoxia and Effect of Ginsenoside Rb1

2016/1/28 16:54:55

Objective To observe pancreatic ER kinase (PERK) signaling of unfolded protein response (UPR). Methods The rats were divided into control, model, and ginsenoside Rb1 (Rb1) groups, and put into hypoxia chamber to establish the acute plateau stress model. The water solutions of Rb1 were given to rats in Rb1 group for 7 d. After that, The behavior of rats was observed by Y-maze and passive avoidance test and the rats were sacrificed in a batch for detection by Western blotting. Results Hypobaric hypoxia mediated UPR pathway accompanied the activation of protective pathways such as PERK-eIF2a-ATF4 and Grp78/Bip pathways. On the other hand, Rb1, the extract from herbal medicine ginseng, increased on the expression of PERK, eIF2a, ATF4, and Grp78/Bip in rats. Conclusion The results indicate that PERK-eIF2a-ATF4-GRP78 pathway is a potential target for therapeutic applications in high altitude diseases and Rb1 can attenuate the injury to memory function caused by hypobaric hypoxia neurotoxicity.

Preparation of Colon-specific and Synchronous Release Pellet Containing Total Alkaloids of Sophora alopecuroides

2016/1/28 16:54:55

Objective To deliver multiple component drugs to colon site and sustain a synchronous release for better therapeutic effect. For achieving this purpose, colon specific pellet containing total alkaloids of Sophora alopecuroides (TASA) was prepared. Methods The pellet was prepared by extrasion-spheronizing and subsequently coated with three layers of two polymers. Results The pellet core consisted of 40% TASA, 1:2 in ratio of Bletilla striata polysaccharide (BSP), an enzyme-degradable material, to microcrystalline cellulose (MCC), filler, and 1% CMC-Na solution as binder by optimization. Concerning of the three coated layers, the outer layer was coated with Eudragit RS30D for controlling drug release in colon, the intermediate layer and the inner layer were coated with same polymer, Eudragit S100, for preventing drug release in upper gastrointestinal tract, which required 23.2%, 21.7%, and 9.3% weigh gain, respectively. The coated pellets released 1.20% of sophoridine and 1.98% of matrine in media mimicking the stomach condition for 2 h, and 23.88% of sophoridine and 22.91% of matrine in media mimicking the intestine for 3 h and finally 90.25% of sophoridine and 89.94% of matrine in colonic conditions within 24 h. And the similarity factor f2 of sophoridine and matrine of release curve for investigated formulation was internal in (50?100) and > 80, demonstrating that sophoridine and matrine in formulation achieved a synchronous release. Conclusion The coated pellets achieve a certain colon-specific release and synchronous release.

LC–MS/MS Method for Quantification of Liquiritigenin in Rat Plasma: Application to Pharmacokinetic Study of Liquiritin

2016/1/28 16:54:55

Objective A simple, sensitive, and rapid LC-MS/MS method has been established and validated for the determination of liquiritigenin (LG) in rat plasma. Methods Naringenin was chosen as internal standard (IS). LG and IS were separated on a Diamonsil C18 analytical column with a mobile phase of methanol-10% methanol in water containing 0.5 mmol/L ammonium formate and 0.2% formic acid (55:45) at the isocratic flow rate of 0.6 mL/min for 10 min. The multiple reaction monitoring (MRM) was performed on a mass spectrometer in the negative ion mode with electro-spray ionization (ESI) source and the transition from precursor ion to product ion was m/z 255.0→119.0 for LG and m/z 271.0→151.0 for IS, respectively. Results The linearity was acceptable in the range of 5-5000 ng/mL (r = 0.9973). The inter-day and intra-day accuracies were in the ranges of ?0.09%?3.25% and ?5.02%?9.21%, respectively. The precision was in the ranges of 3.60%?12.4% and 0.909%?6.89%, respectively. LG was stable in the course of analysis and storage. Conclusion The LC-MS/MS method was successfully applied to the pharmacokinetic study for the first time in rats after ig and iv administration of liquiritin (LQ), a glycoside of LG, at pharmacologically effective levels.

Hemolytic assay for Huangqi Injection

Hang Xiao,Yu-bin Xu,Xia Liu and De-qiang Dou — 2016/1/28 16:54:55

Objective Huangqi Injection is a preparation with an extract of Astragali Radix which has a long history of being used as a tonic to strengthen the body’s immunity. Anaphylaxis and hemolysis are two main adverse drug reaction (ADR) of injections. Our study was aimed to establish an approach for the (ADR) prediagnosis of Huangqi Injection. Methods An in vitro model for anaphylactoid assay of Huangqi Injection based on the release rate of histamine and β-hexosaminidase of RBL-2H3 cells induced by injections and a colorimetric method based on the detection of hemoglobin resulted in the erythrocyte hemolysis for prediagnostic assaying the hemolytic ADR of injections were established. Results Both histamine and β-hexosaminidase are the anaphylactiod mediators, but β-hexosaminidase release induced by Huangqi Injection could not be determined by spectrophotometry due to the interference of the injection itself. In addition, normal hemolysis and abnormal hemolysis were discovered during the experiment. The fingerprints and tannins in different batches of injections showed obvious differences, indicating that the content of tannins was related to abnormal hemolysis and higher histamine-secreting from RBL-2H3 cells. Conclusion The results indicate that the hemolytic assaying method is not only suitable for prediagnostic assaying of hemolytic ADR of herbal medicine injection, but also partly reflects the anaphylaxis of herbal injections, and tannins may be the major factors causing abnormal hemolysis.

Molecular Cloning and Expression of Squalene Epoxidase from a Medicinal Plant, Bupleurum chinense

Ke Gao,Jie-sen Xu,Jing Sun,Yan-hong Xu,Jian-he Wei and Chun Sui — 2016/1/28 16:54:55

Objective In plant, squalene epoxidase (SE) catalyzes the first oxygenation step in the biosynthetic pathway of triterpenoid and phytosterol, representing one of the rate-limiting enzymes in this pathway. Bupleurum chinense is an important medicinal herb with its major active constituents such as triterpenoid saponins and saikosaponins. In order to obtain the series of enzymatic genes involved in saikosaponin biosynthesis, a cDNA of SE, designated BcSE1, was cloned from B. chinense. Methods The BcSE1 gene was cloned by homology-based PCR and 5’/3’ RACE methods from the adventitious roots of B. chinense. The physical and chemical parameters of BcSE1 protein were predicted by protparam. In order to discover hints in amino acid sequences on the dominant functions in the biosynthesis of saponin or phytosterol, sequences of SE from other plants were downloaded from NCBI for sequences alignment and phylogenetic analysis. BcSE1 was cloned into a yeast mutant KLN1 (MATa, erg1::URA3, leu2, ura3, and trp1) to verify the enzyme activity of BcSE1. Additionally, the tissue-specific expression and methyl jasmonate (MeJA) inducibility of BcSE1 were investigated using quantitative real-time PCR. Results The predicted protein of BcSE1 is highly similar to SEs from other plants sharing amino acid sequence identities of up to 88%. The BcSE1 can functionally complement with yeast SE gene (ERG1) when expressed in the KLN1 mutant (MATa, erg1::URA3, leu2, ura3, and trp1). Using as controls with β-amyrin synthase (β-AS) which is presumed to catalyze the first committed step in saikosaponin biosynthesis and a cycloartenol synthase (CAS) relating to the phytosterol biosynthesis, the transcript of BcSE1 was significantly elevated by MeJA in adventitious roots of B. chinense and the transcript of BcSE1 was most abundant in the fruits and flowers of plants, followed by that in the leaves and roots, and least in stems. Conclusion It is the first time to illustrate the molecular information of SE in B. chinense and to clone the full-length SE gene in plants of genus Bupleurum L.

Isolation and Chemotaxonomic Significance of Chemical Constituents from Rubus parvifolius

2016/1/28 16:54:55

Objective To study the chemical constituents from the roots of Rubus parvifolius. Methods The chemical constituents were extracted and purified by silica gel column chromatography. NMR spectra were used for structural identification. Results Phytochemical study on the roots of R. parvifolius led to the isolation of one ceramide (1), two anthraquinones (2 and 3), four triterpenoids (4-7), two flavonoids (8 and 9), one fatty acid ester (10), and two sterols (11 and 12). Conclusion Compound 1 is isolated from the plants of family Rosaceae for the first time, and compounds 2-5 are isolated from genus Rubus for the first time. Though R. parvifolius shares the same major chemical types (triterpenoid, flavonoid, and anthraquinone) with those of R. alceaefolius, a substituent of R. parvifolius, their individual constituents are different. In addition, R. parvifolius contains ceramide (1) with high concentration, while caffeoylquinic acid reported in R. alceaefolius has not been found in R. parvifolius. Furthermore, the results from our phytochemical study are consistent with the DNA phylogenic relationship between R. parvifolius and R. alceaefolius (two separated subgenera), suggesting that the substitution of the former species with the latter one in folk medicine might not be suitable.

Chemical Constituents from Leaves of Camellia nitidissima and Their Potential Cytotoxicity on SGC7901 Cells

2016/1/28 16:54:55

Objective To isolate and identify the bioactive phytochemicals from the leaves of Camellia nitidissima. Methods The chemical constituents were isolated and purified by repeated silica gel, Sephadex LH-20, MCI gel columns, recrystallization， and semi-preparative HPLC techniques. The chemicl structures of these compounds were identified on the basis of spectral data including NMR and MS. Then quorum sensing inhibition (QSI) activities of these compounds were tested using Chromobacterium violaceum CV026 as the bioindicator strain. The antitumor activities of these compounds were measured using SGC7901 as cell proliferation and cytotoxicity. Results α-Spinasteryl-β-D-glucopyranoside (1), stigmasta-7,22-diene-3-O-[α-L-arabinopyranosyl (1→2)]-β-D-galactopyranoside (2), kaempferol 3-O-[2-O-(trans-p-coumaroyl)-3-O- α-D-glucopyranosyl]-α-D-glucopyranoside (3), aromadendrin (4), catechin (5), phlorizin 4′-O-β-D-glucopyranoside (6), (3R,6R,7E)-3-hydroxy-4,7-megastigmadien- 9-one (7), dodecanoic acid (8), 3β-acetoxy-20-lupanol (9), and 3β,6α,13β- trihydroxyolean- 7-one (10) were successively isolated from the leaves of C. nitidissima. Unfortunately, these compounds had no QSI activity. Based on Cell Counting Kit-8 (CCK-8) assay, compound 10 showed the best anti-tumor activity of all compounds (IC50 = 91.7 μg/mL). Conclusion Apart from compounds 4 and 5, other eight compounds are reported in this plant for the first time. All compounds show no QSI activity, compound 10 shows potential cytotoxic activity on SGC7901 cells in vitro.

Flavonoids from Heartwood of Dalbergia cochinchinensis

2016/1/28 16:54:55

Objective To study the flavonoids from the heartwood of Dalbergia cochinchinensis. Methods The chemical constituents were isolated and purified by combination of silica gel, macroporous resin, Sephadex LH-20, and ODS column chromatography. Their structures were identified by means of spectral analysis. Results Fifteen flavonoids were isolated and identified as pinocembrin (1), liquiritigenin (2), galangin (3), 7-hydroxy- 6-methoxyflavone (4), naringenin (5), alpinetin (6), 2,3-dimethoxyxanthone (7), 6,4′-dihydroxy-7-methoxy-flavan (8), mucronulatol (9), 7,8-dihydroxyflavanone (10), 5,7,3′,5′-tetrahydroxyflavanone (11), 4,2′,5′-trihydroxy-4′-methoxychalcone (12), isoliquiritigenin (13), butein (14), and 3′,5′,5,7-tetrahydroxy-6-C-β-D-glucopyranosyl- flavanone (15), respectively. Conclusion Compounds 7, 8, 10, and 15 are isolated from the plants of Dalbergia L. f. for the first time, and compounds 1, 3, 5, 6, 9, 11, 12, and 14 are isolated from this plant for the first time.

Research on Quality Markers of Moutan Cortex: Quality Evaluation and Quality Standards of Moutan Cortex

2017/10/17 0:00:00

Objective To identify the quality markers of Moutan Cortex (MC) and establish the quality evaluation methods for multi-component assay and fingerprinting of MC. Methods The chemical constituents in MC were identified by HPLC-QTOF-MS. UPLC was employed for the multi-component assay and fingerprinting of MC. Furthermore, text mining was carried out to review the biosynthesis pathways and pharmacological and pharmacokinetic studies related to MC, and in silico target fishing was conducted to construct compound-target networks for MC. Results Sixteen compounds were clearly identified in MC and their structures were confirmed through comparison with literature data. In addition, the biosynthetic pathways and component specificities of the identified compounds were summarized and confirmed by text mining. Pharmacological activities, including traditional usage and modern pharmacological studies were summarized. A total of 282 targets from Homo sapiens were fished for 13 compounds. In addition, pharmacokinetic studies of different compounds were synopsized. Finally, multi-component assay and fingerprint of MC were established. Conclusion Eight major components are selected as quality markers of MC, such as oxypaeoniflorin, apiopaeonoside, albiflorin, paeonolide, paeoniflorin, 1,2,3,4,6- penta-O-galloyl-β-D-glucose, mudanpioside C and paeonol. These eight quality markers are successfully applied to the quality evaluation of MC, and could be useful in improving the current quality standards of MC.

Tyrosinase Inhibitory Activity of Total Triterpenes and Poricoic Acid A Isolated from Poria cocos

Shu-hua Hu, Gao Zhou and You-wei Wang — 2017/10/17 0:00:00

Objective With the improvement of people’s living standards, people’s requests for beauty are increasing. Skin whitening and lightening have become the pursuit of many women, and whitening and removing freckles have become the focus of scientific research. At present, widely used whitening agents, such as kojic acid, vitamin C, and its derivatives, have shortcomings such as poor stability and retarded effect. Therefore, safer and more effective whitening products from herbs are urgently needed. To explore the possibility of triterpenes as whitening active substance, the effects of total triterpenes of Poria (TTP) and poricoic acid A (PAA) on mushroom tyrosinase activities and B16 cells were investigated, and their mechanisms on mushroom tyrosinase were also studied. Methods Using arbutin and nicotinamide as reference substances, we determinated the inhibitory effects of TTP and PAA on mushroom tyrosinase and tyrosinase in B16 cells and then studied the inhibitory mechanism on mushroom tyrosinase. Results TTP and PAA exhibited good inhibitory effects on the activities of monophenolase and diphenolase in mushroom tyrosinase, as well as a certain inhibitory effect on tyrosinase in B16 cells. Conclusion TTP and PAA are potential whitening active ingredients.

Variation of Sweet Chemicals in Different Ripening Stages of Wolfberry Fruits

Dan Qian, Jian Yang, Li-ping Kang, Rui-feng Ji and Lu-qi Huang — 2017/10/17 0:00:00

Objective To evaluate the variation of sweet chemicals (sugars and betaine) in fruits of six species in genus Lycium L. i.e. Lycium truncatum, L. cylindricum, L. dasystemum, L. dasystemum var. rubricaulium, L. chinense, and L. barbarum harvested at three different ripening stages. Methods A simple and effective method based on UPLC-ELSD was developed for the simultaneous determination of two monosaccharides (glucose and fructose), two disaccharides (sucrose and maltose), one sugar alcohol (xylitol), and betaine in wolfberry fruits (goji berries) of genus Lycium L. Results The six species of wolfberry fruits harvested at the three different ripening stages were evaluated in sugars and betaine contents. Fructose and glucose were the predominant sugars in mature wolfberry fruits. Fructose, glucose, and betaine, as well as total sugar contents, increased continually over the ripening process. L. truncatum and L. dasystemum had higher contents of sugars and betaine than the other species. Conclusion UPLC-ELSD is a simple, reliable and effective method for analysis of the sweet chemicals in wolfberries. Wolfberry fruits at the different ripening stages were significant different in sweetness. L. truncatum and L. dasystemum could be the potential sources of wolfberry fruit sources.

Protective Effect of Salvianolic Acid A on Brain Endothelial Cells after Treatment with Deprivation and Reperfusion of Oxygen-glucose

2017/10/17 0:00:00

Objective Salvianolic acid A (SAA) has a significant protective effect on ischemia/reperfusion injury of brain. However, it is not clear for SAA to exert its cerebral protection by targeting at the microvascular endothelial cells of blood brain barrier (BBB). Our previous study demonstrated that SAA could hardly pass through the BBB. This present study was therefore designed to investigate the protective effect of SAA on brain microvascular endothelial cells (BMECs) induced by deprivation and reperfusion with oxygen-glucose. Methods Rat BMECs were treated with oxygen glucose deprivation (OGD), followed by reperfusion (OGD/R). Cell viability was assessed by MTT and the content of reactive oxygen species (ROS) in cells after OGD/R in the absence or presence of SAA. GC-MS based metabolomic platform was applied to evaluate the regulation of SAA on the cellular metabolic perturbation induced by OGD/R. Results OGD/R significantly increased the production of intracellular reactive oxygen species (ROS), and decreased the activity of cells. SAA significantly reduced ROS and improve the cell viability. Metabolomic study revealed distinct perturbation of metabolic pathways of energy metabolism in the BMEC induced by OGD/R, while SAA significantly regulated the perturbed metabolism involved in energy metabolism pathways, especially for intermediates in TCA cycle. Conclusion SAA shows protective effects on BMECs involved in central nervous system.

Amelioration of Tripterygium wilfordii Polycoride on TNBS/Ethanol-induced Ulcerative Colitis via Inhibiting Lipid Peroxidation and Its Downstream Inflammatory Meditors

Yi-jun Zhou, Dan-ping Qin, Yao-dong Wang, Qiang Yang, Chun-li Zhang4 and Qun Dai — 2017/10/17 0:00:00

Objective To study the protective effect of Tripterygium wilfordii polycoride (TWP) against TNBS/ethanol-induced ulcerative colitis (UC) rat model. Methods TNBS and ethanol enema were adopted to build TNBS/ethanol-induced UC rat model. Ninety male Wistar rats were divided into six groups: normal, model, low-, medium-, high-dose TWP and azathioprine (AZA) groups, each for 15 rats. All rats were administered by corresponding medicine for 14 d. After 14 d, corresponding colon tissues underwent general and microscopic evaluation. Blood samples were taken from heart and serum was separated by centrifugation. MDA, SOD, GSH, IL-1β and TNF-α levels in serum were tested by ELISA. Colonic samples underwent RT-PCR and Western blotting analysis. Results DAI, general and microscopic evaluation all showed that TWP could promote colonic mucosa healing and such effect was equal to AZA. ELISA results about lipid peroxidation indicated that TWP could decrease MDA level and increase SOD and GSH levels in a dose-dependent manner. TWP with high dose could strongly decrease the MDA level and increase the SOD and GSH levels (P ＜ 0.01). ELISA results about inflammatory cytokines indicated that TWP could inhibit the expression of IL-1β and TNF-α in a dose-dependent manner. Western blotting analysis and RT-PCR all indicated that no matter in mRNA level or protein level, TWP could inhibit the expression of NF-κB, TNF-α, IL-1β, and IFN-γ in a dose-dependent manner. The inhibitory effect of AZA towards NF-κB was slightly weaker than TWP with high dose (P ＞ 0.05), whereas slightly stronger towards terminal inflammatory cytokines (P ＞ 0.05). Conclusion TWP could significantly lower the infiltration of inflammatory cells under microscope, eventually led to mucosa healing, the mechanism of which was to inhibit lipid peroxidation, then further inhibit NF-κB activation, eventually lower the expression of inflammatory meditors locally and systemically.

Synergistic Antimicrobial Activity of Berberine Hydrochloride, Baicalein and Borneol against Candida albicans

Yan Liu, Yu Zhao, Da-liang Guo, Wen-wei Liu and Yu-xuan Liu — 2017/10/17 0:00:00

Objective To determine the synergistic effects of berberine hydrochloride, baicalein, and borneol in different combinations on Candida albicans. Methods The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the three agents, and the checkerboard method was simultaneously used to determine the MIC and fractional inhibitory concentration index (FICI) of the combination of three antimicrobial agents to study their extracorporeal effects. Results Berberine hydrochloride was the most potent inhibitor of C. albicans (MIC and MBC of 0.160 and 0.640 mg/mL), followed by borneol (MIC and MBC of 0.320 and 0.640 mg/mL) and baicalein (MIC and MBC of 1.28 and 20.48 mg/mL). Moreover, the antifungal effect of the combination was significantly stronger than that tested alone. Further in vivo study showed that the mortality rate of tainted mice reduced over 50% compared with the control group. Conclusion The results of experiments in vitro and in vivo indicate the synergistic effect of the combination of three antimicrobial agents on C. albicans, which can make reference for the future clinical treatment.

Anti-inflammatory and Anti-oxidant Effects of Licorice Flavonoids on Ulcerative Colitis in Mouse Model

Dong-yu Liu, Li Gao, Juan Zhang, Xiao-wei Huo, Hui Ni and Li Cao — 2017/10/17 0:00:00

Licorice (Glycyrrhizae Radix or Liquiritiae Radix) is traditionally used to treat various diseases including inflammation and gastric ulcers. Licorice is rich in flavonoid compounds and possesses anti-inflammatory activities. To investigate the protective effects of licorice flavonoids (LFs) in both acetic acid-induced and dextran sulphate sodium (DSS)-induced ulcerative colitis (UC) mouse model and its underlying mechanism. Acute UC was induced by intra-rectal acetic acid (4% v/v) after pretreatment with LFs (100, 200, and 400 mg/kg, p.o.), 0.9% saline (20 mL/kg, p.o.) or Sulfasalazine (SASP) (600 mg/kg, p.o.) for 10 d. Quantitative analysis of chemical components of LFs was also conducted by HPLC. Our results showed that pre-treatment with LFs significantly reduced the wet weight/length ratio of colon, percentage of affected area, macroscopic and histological damage scores in acid-induced UC mice. LFs also significantly decreased the oxidative stress and pro-inflammatory cytokines, upregulated nuclear factor erythroid 2-related factor 2 (Nrf2) pathway and downregulated nuclear transcription factor kappa B (NF-κB) pathway. At last, LFs also showed obvious antiulcer effect on the DSS-induced UC model. The major components of LFs were licochalcone A, glabrone, licoflavone, and licoflavone B. This study demonstrates that the protective effect of LFs may at least in part be due to its anti-oxidant activity through Nrf2 pathway and anti-inflammatory activity through NF-κB pathway.

Metabolomic Analysis of Biochemical Changes in Urine of Osteoarthritis Rat and Interventional Effects of Bushen-Huoxue Herb Couple

Hua Yin, Ling Wang, Qing-lin Li, Jian-hua Zhang, Lin Zhang and Xuan Wang — 2017/10/17 0:00:00

Objective Osteoarthritis (OA) is a kind of degenerative arthritis, involving degradation of joints, including articular cartilage and subchondral bone. Bushen-Huoxue Herb Couple (BHHC) is a Chinese herb pair with invigorating the kidney and promoting blood circulation. In our study, a Gas Chromatography-Mass Spectrometer (GC-MS) based metabolomics was developed to achieve a systematic observation of the progression of OA and evaluate the efficacy of BHHC in OA rats. Methods Forty Sprague-Dawley rats (half males and half females) were randomly divided into five groups, including normal group, OA model group, sham-operated group, BHHC group, positive medicine (PM) group after anterior cruciate ligament reconstruction and meniscectomy on rats. Urine samples for analysis were collected on week 0, 2, 4, 6, 8, 10, 12, and 14 during the experiment. Results The histological results showed that BHHC had a strong action of repairment and reconstruction of joint damage. By combining with partial least squares discriminant analysis (PLS-DA), 14 biomarkers were identified in urine samples. The biomarkers were mainly involved 10 metabolism pathways, such as tricarboxylic acid cycle metabolism, acid amino metabolism, tryptophan metabolism, lipid metabolism and phenylalanine metabolism. Using these potential biomarkers as a screening index, it indicated that BHHC could reverse the pathological process of OA through regulating amino acid metabolism, lipid metabolism, nucleotide metabolism and energy metabolism. Conclusions This study demonstrates that a metabolomics is a useful tool for identifying potential OA biomarkers and investigating the underlying mechanisms of Traditional Chinese Medicine in OA treatment.

Determination and Pharmacokinetic Study of Nitidine Chloride in Rat Plasma after Intragastrical Administration by LC-ESI-MS/MS Method

Dong-mei Wang, Jing-fan Zhou, Xiao-bin Zhong and Jie Feng — 2017/10/17 0:00:00

Objective To study the pharmacokinetics of nitidine chloride (NC) in rat plasma after intragastrical (i.g.) administration. Methods A liquid chromatography-electrospray ionization-mass/mass sprectrometry (LC-ESI-MS/MS) was used and carbamazepine was used as an intermal standard (I.S.). The rat plasma samples were deproteinized with acetonitrile and the resultant supernatant was assayed on an analytical DiamonsilTM ODS C18 column (2.1 mm × 150 mm) equipped with a C18 guard column (4 mm × 20 mm) with a mobile phase of acetonitrile–10 mM ammonium acetate buffer–formic acid (35: 65: 0.2, v/v/v) at the flow rate of 0.25 mL/min. The LC–MS was carried out on a triple-quadrupole mass spectrometry equipped with an ESI and positive selected-ion monitoring. Target ions were monitored at [M - Cl]+ m/z 348.2 for NC and [M + H]+ m/z 237.2 for I.S., respectively. Results The simple one step deproteinize and rapid analysis method were successfully used in pharmacokinetic study on NC after i.g. administration. The linear relationship was good over the range of 2.5 – 1000.0 ng/ml (r2 = 0.999 2) in rat plasma. The lower limit of quantification and detection were 2.5 and 1.6 ng/ml, respectively. The extraction recovery was in the range of 86.54 – 98.60%. The intra- and inter-day precisions (relative standard deviation) were less than 6.00%, with accuracies deviation between 89.40 to 95.57%. A two-compartment pharmacokinetic open model was proposed and validated to explain the apparent biphasic disposition of NC in rat plasma after i.g. administration. Conclusion This study was successfully applied to a pharmacokinetic study of NC in rats plasma following i.g. administration and could be used for preclinical and clinical pharmacokinetic evaluation of NC.

Protective Effects and Underlying Mechanisms of Compound Herba Houttuyniae in db/db Mice

Yun Fang and Hai-ying Wang — 2017/10/17 0:00:00

Objective Houttuynia cordata Thunb and Fructus Arctii are two traditional Chinese medicine (TCM) compounds widely used for reducing heat and toxins in the body. In addition, Fructus Arctii has been used in clinical therapy for diabetic nephropathy. On the basis of the TCM theory of heat in diabetes mellitus and the clinical efficacy of Fructus Arctii, the two compounds were used to develop a formula named Compound Herba Houttuyniae. This study evaluated the renoprotective effects of different Compound Herba Houttuyniae extracts and assessed their mechanisms through the Janus kinase/signal transducer and activator of transcription (JAK/STAT)–suppressors of cytokine signaling (SOCS)-1 pathway. Methods The normal group comprised db/m mice (n = 8); db/db mice were randomly divided into six groups according to the applied treatment method: model group (no treatment), AG490 group, petroleum ether extract (PEE) group, ethyl acetate extract (EAE) group, N-butanol extract (NE) group, and water extract (WE) group (n = 6 in each group). The general status, biochemical indicators, and renal histological changes in the mice were evaluated, and the JAK/STAT–SOCS-1 pathway was assessed. Results The NE and WE groups showed a decrease in 24-h urinary protein and albumin levels as well as serum uric acid and fibronectin levels but showed no changes in fasting plasma glucose and hemoglobin A1c levels. The protein expression of JAK2, STAT3, P-JAK2, and P-STAT3 and the gene expression of JAK2 and STAT3 in kidney tissues were significantly increased (P < 0.05) in the model group. Compared with the model group, the expression of P-JAK2 and P-STAT3 was downregulated in the extract treatment groups. The expression of SOCS-1 was increased in the NE and WE groups (P < 0.05). However, the expression of JAK2, STAT3, JAK2mRNA, and STAT3mRNA in the treatment groups did not show any significant differences. Conclusion Compound Herba Houttuyniae showed renoprotective effects in db/db mice, and the strongest effects were observed in the WE and NE groups. The underlying mechanism for these effects might involve the regulation of the JAK/STAT–SOCS-1 pathway.

Simultaneous Determination of 10 Anthraquinones in Rhubarb Based on HPLC-Q-HR/MS

2017/10/17 0:00:00

Objective To develop a simple, sensitive, and precise method for simultaneous determination of 10 anthraquinones in Rhubarb. Methods HPLC-Q-HR/MS was employed for simultaneous quantification of free anthraquinones (aloe-emodin, emodin, chrysophanol, physcion, and rhein) and their glycosides. Chromatographic analysis was performed on an XBridgeTM C18 column (2.1 mm × 150 mm, 5 μm) with mobile phases consisting of 3 mmol/L ammonium acetate (A) and methanol (B) at a flow rate of 0.3 mL/min. Results All calibration curves exhibited good linear relationship (R2 > 0.999). The limits of detection (LOD) and quantification (LOQ) were in the range of 0.39-2.97 ng/mL and 0.56-8.90 ng/mL, respectively. The overall intra- and inter-day precisions of analytes presented as relative standard deviations (RSDs) were less than 2.79%. Relative recoveries varied between 97.83% and 104.28%. The validated method was applied to assess the quality of Rhubarb collected from different regions of China. Results showed that chrysophanol and rhein-8-O-β-D-glucoside was the largest portion of free anthraquinones and anthraquinone glycosides in Rhubarb, respectively. The total content of anthraquinones was higher in Rhubarbs from Sichuan, Qinghai, Yunnan, and Gansu provinces than that in those from Shandong and Henan provinces, while no significant variability existed in different regions of the same province. Conclusion HPLC-Q-HR/MS method is accurate and reliable for simultaneous quantification of above free anthraquinones and their glycosides in Rhubarb and can be applied to standardize the quality of Rhubarb and its quality control in different regions.

Authenticity Survey of Cuscutae Semen on Markets Using DNA Barcoding

Zi-tong Gao,Li-li Wang,Xiao-yue Wang,Yang Liu and Jian-ping Han — 2017/8/9 0:00:00

Objective Using authentic raw herbal materials is fundamental to herbal medicine quality. Cuscuta chinensis and C. australis are two important species of Cuscutae Semen recorded in Chinese Pharmacopoeia. Due to having tiny bodies of seeds, it is extremely difficult to differentiate them from adulterants and closely related species by morphologic characteristics, leading to serious safety problems. Methods In this study, we developed a fast and efficient method to identify Cuscutae Semen on the market. First, a total of 207 ITS2 sequences representing 45 related species of Cuscutae Semen were collected to construct a standard DNA barcode database, then 33 commercial samples purchased from markets were analyzed by BLAST, and Neighbor-joining tree was used to verify the efficacy of the database. Results The percentage of counterfeits and adulterants in the 33 commercial samples were up to 69.7%, and only 10 commercial products were found to be genuine. The adulterated species included 11 species (Amaranthus hybridus, Brassica carinata, Brassica juncea var. megarrhiza, Chenopodium album, Corispermum heptapotamicum, Cuscuta alata, Cuscuta japonica, Cuscuta monogyna, Foeniculum vulgare, Glycine max, and Medicago sativa). Conclusion DNA barcoding is a fast and efficient method to identify Cuscutae Semen on the market.

Validation of Reliable Reference Genes for Accurate Normalization in RT-qPCR Analysis of Codonopsis pilosula

Ling-ya Cao 1,Xiao-xiao Li 2,Dong Wang2,Hai-feng Sun3 and Jian-ping Gao2 — 2017/8/9 0:00:00

Objective To identify reliable reference genes (RGs) for normalization of real-time PCR (RT-qPCR) data in Codonopsis pilosula. Methods The expression profiles of 10 candidate RGs (GAPDH, ACT, α-TUB, β-TUB, UBQ, CYP, EF-1α, NAC, F-box and PP2A) were examined in C. pilosula during the phenological period. The raw materials examined included roots, stems, leaves, and flower buds at flowering and boll-forming stages, five growth stages of untreated and treated roots with plant growth retardant. The best-suited RGs were accessed using geNorm, NormFinder, BestKeeper, and RefFinder algorithms. Results The best-ranked references genes differed across the samples. GAPDH and PP2A were the most suitable for expression analysis in untreated tissues while GAPDH, α-TUB, and PP2A were ranked as the three most stably expressed genes in untreated roots, while NAC and CYP were the most stably expressed genes in stressed (i.e., treated) roots. The expression of UGPase, a key enzyme for CPP biosynthesis, was determined to further validate the selected RGs. Conclusion A total of 10 RGs can be used as reference genes of C. pilosula, however the appropriate one should be used as it may chance.

Sensitive Analysis and Pharmacokinetic Study of Berberrubine Using LC-MS/MS

2017/8/9 0:00:00

Objective To develop a sensitive and reproducible liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to evaluate the pharmacokinetic behavior of berberrubine (BRB) and its glucuronide (BRBG) in rats. Methods BRB, BRBG and tetrahydroberberine (THB, internal standard) were isolated by liquid-liquid extraction in rat biological samples. Chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C18 (2.1 mm × 50 mm, 3.5-Micron) with a gradient mobile phases primarily containing acetonitrile, water with 0.1% formic acid and 5 mm ammonium acetate. The analytes were monitored by MS/MS in positive electrospray ionization mode. Herein, the feasibility of new developed method was validated with respect to specificity, linearity, precision, accuracy, stability, extraction efficiency and matrix effect. The appropriate method was used for the pharmacokinetic study in rats. Results The new developed method could be applied to the pharmacokinetic study of BRB in rats. BRB and BRBG showed good linearity over the ranges of 2-1000 ng/mL and 5-2000 ng/mL, respectively, and precision was no more than 15%. The accuracy, specificity and stability could be acceptable. Conclusion The new method is sensitive and reproducible. In pharmacokinetic study, BRB showed nonlinear elimination property. Meanwhile, BRB was rapidly absorbed and widely distributed in various tissues with the highest exposure of BRB in kidney and liver. The absolute bioavailability of BRB was determined to be 8.2% and at the dose of 40 mg/kg, a total of 44% BRB was excreted in urine and feces.

Rapid Determination of Total Content of Five Major Anthraquinones in Rhei Radix et Rhizoma by NIR Spectroscopy

2017/8/9 0:00:00

Objective To establish a new method with near-infrared (NIR) spectroscopy to determine the total content of emodin, chrysophanol, rhein, aloeemodin, and physcion. NIR was used in this study to provide rapid and nondestructive analysis results. Methods In the first place, HPLC was used to measure the total content of emodin, chrysophanol, rhein, aloe-emodin and physcion in Rhei Radix et Rhizoma (RR) as a reference. In the second place, the spectral regions, regression methods, pretreatment methods, and partial least squares (PLS) factors were compared to increase the feasibility of the model. In the last, the root mean square error of calibration (RMSEC), root mean square error of cross validation (RMSECV), root mean square error of prediction (RMSEP), and correlation coefficient (r ) were used as assessment parameters. Results PLS with first derivative pretreatment in the ranges of 4242-5581 cm-1, 5885-6233 cm-1 and 6394-7011 cm-1 provided the best results. The RMSEC and RMSEP obtained were 0.134 and 0.226 respectively. The according determination coefficients of the quantitative model were 0.99 and 0.94. Conclusion NIR spectroscopy as a quick and nondestructive analytical method may be used to determine the total content of emodin, chrysophanol, rhein, aloeemodin, and physcion for the quality control of RR.

UFLC-DAD-ESI-IT-TOFMSn Analysis on Biotransformation of Tongmai Formula Incubated with Human Intestinal Bacteria

Shuai Wu,Qi-le Zhou and Xiu-wei Yang — 2017/8/9 0:00:00

Objective To investigate the biotransformation of Tongmai formula (TMF) in incubated system of human intestinal flora (HIF). Methods The technique of ultra fast liquid chromatography with diode array detector and coupled with electrospray ionization ion trap time-of-flight multistage mass spectrometry (UFLC-DAD-ESI- IT-TOFMSn) was adopted to determine the products of TMF biotransformed by HIF. Results Totally 66 constituents were detected and identified according to the accurate mass measurements (< 5 ppm) and effective MSn fragment ions. Meanwhile, the potential biotransformational pathways of compounds in TMF transformed by HIF were firstly proposed. Desugarization, hydroxylation, and methylation were the major reactions in the biotransformation mechanism of TMF by HIF. Conclusion This study will be helpful to clarify the material basis of pharmacological activities from TMF in vivo.

Simultaneous Determination of Saponins in Dripping Pills Made from Astragali Radix and Panax notoginseng by UPLC-ELSD

Jing-jing Peng1,Dong-xiang Li2,Jing-yi Huang2,Ling Tong2 and Bo-yang Yu1 — 2017/8/9 0:00:00

Objective To develop a simple and fast method for removing polyethylene glycol (PEG) and simultaneous determination of fives saponins, i.e. astragaloside IV, noto- ginsenoside R1, ginsenoside Rg1, ginsenoside Rb1, and ginsenoside Rd in dripping pills made from Astragali Radix and Panax notoginseng. Methods The extraction method was based on a liquid-liquid extraction using water-saturated n-butanol and the quantitative determination was based on ultra-performance liquid chromatography coupled with evaporative light scattering detection (UPLC-ELSD). The chromatographic analysis was performed on an Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) with a gradient elution of acetonitrile-0.1% formic acid aqueous solution within a runtime of 15 min. Results Compared to different methods, the proposed method could remove the interference of PEG in formulation. And the calibration curves showed good linearity during the test ranges. The method was validated for limits of detection and quantification, precision, and reproducibility. The recoveries were within the range of 96.87% - 99.97%. In addition, the verified method was firstly applied to determination of the five active ingredients in Qishen Yiqi Dripping Pills (QYDP) simultaneously. Conclusion The contents of five active ingredients are stable and homogeneous in QYDP, which indicates that the method could be readily utilized as a quality evaluation method for this traditional Chinese medicine dripping pills made from Astragali Radix and Panax notoginseng.

Comprehensive Evaluation on Effect of IMPX977 on Expression of Methyl-CpG-binding Protein 2 in Rats

2017/8/9 0:00:00

Objective To investigate the effect of IMPX977 on methyl-CpG-binding protein 2 (MeCP2) expression in rats. Methods Forty-eight SD rats were randomly divided into four groups: normal control group, olive oil (negative control, 5 mL/kg oil) group, and 10 mg/kg and 30 mg/kg IMPX977 administration groups. All rats were given corresponding dose of drugs each other day and administered orally for two weeks. Tissues including cortex and cerebellum were collected from rats to assay the expression of MeCP2 by quantitative RT-PCR and Western blotting. Results The IMPX977 supplement showed no significant effect on the body weight of rats. In normal rats, MeCP2 was highly expressed in cerebellum, cortex and hippocampus, and less expressed in heart, spleen and lung. In addition to male rats, compared with the control group, the expression of MeCP2 mRNA was significantly increased in cerebellum after 30 mg/kg IMPX977 treatment and contrarily, absolutely decreased in cortex of all treatment groups. Furthermore, in female rats MeCP2 mRNA was reduced in cortex of both olive oil and 30 mg/kg IMPX977 treatment groups compared with control group. Meanwhile, MeCP2 protein level was significantly elevated in cerebellum of treated male rats compared to the control group. In contrast to the control group, the expression of MeCP2 protein in both cerebellum and cortex of female rats in other three groups was increased. Conclusion IMPX977 treatment (10 mg/kg) may elevate the expression of MeCP2, which establishes experimental foundation for the further research on rat models of Rett syndrome.

Protective Effects of Hydroxysafflor Yellow A against Oxidative Damage of β-Mercaptoethanol During Neural Differentiation of Mesenchymal Stem Cells

Xiao-qing Song1,Li-ning Su1,Hui-ping Wei1,Ying-hui Liu2 and Hai-feng Yin1 — 2017/8/9 0:00:00

Objective To study the protective effects of hydroxysafflor yellow A (HSYA) against the oxidative damage caused by β-mercaptoethanol (BME) during neural differentiation of mesenchymal stem cells (MSCs) in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. G1: normal group which was cultured using basic medium (DMEM containing 10% FBS all the time); G2: unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium (DMEM containing 10% FBS and 1 mmol/L BME); G3: protected group which was firstly cultured using protective medium (DMEM containing 10% FBS and 160 mg/L HSYA) for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.

Phytochemical, Anti-oxidant, Larvicidal, and Antimicrobial Activities of Castor (Ricinus communis) Synthesized Silver Nanoparticles

Namita Soni and Ramesh C. Dhiman — 2017/8/9 0:00:00

Objective Castor possesses the medicinal properties. Hence, castor extract has the reducing power agent was used in nanoparticles synthesis. Extract of castor also produces secondary metabolites (phenolics, reducing agent) and exhibits anti-oxidant property, which was used as larvicide and antimicrobial agent. In the present study the silver nanoparticles (AgNPs) were synthesized using aqueous leaf extract of castor. Methods The synthesized AgNPs were characterized with UV-visible spectroscopy and transmission electron microscopy (TEM). The phenolics, reducing sugar and total sugar were evaluated performing Folin-Ciocalteu reagent method, DNS method and Phenol-sulphuric acid method. Anti-oxidant activity of synthesized nanoparticles was evaluated using DPPH assay. The larvicidal activity of synthesized AgNPs was evaluated against the third instar of Anopheles stephensi and Aedes aegypti. The mortality was observed after 24 h by probit analysis. Further, the antimicrobial activity was reported against Staphylococcus aureus (Gram positive) and Pseudomonas aeruginosa (Gram negative). Results The UV spectra showed maximum absorbance at 445 nm and TEM analysis indicated spherical shape of nanoparticles with average size of 8.96 nm. The synthesized nanoparticles also possessed anti-oxidant potentials. Thus, it could be used as potential free radical scavenger. The larvae of A. stephensi were found more susceptible to AgNPs than larvae of A. aegypti. Conclusion Due to the medicinal properties and reducing power activity, in the present study AgNPs have been synthesized using the aqueous leaves extract of castor and access their phytochemical, anti-oxidant, larvicidal and antimicrobial properties.

Inhibition of Cytochrome P450 by Nomilin and Obacunone and Potential Mechanism in Human Liver Microsomes

2017/8/9 0:00:00

Objective Nomilin and obacunone are two important limonoids that are well known for their anticancer effect. Previous studies showed that limonoids had inhibitory effect on cytochrome P450 3A4 (CYP3A4). However these effects are inconclusive with regards to prediction of potential drug interactions. Methods Nomilin or obacunone was pre-incubated with HLMs for 30 min. Following 10-fold dilution from the pre-incubation concentration, a second incubation was performed in the presence of NADPH and cytochrome P450 substrates for 15 min. The reaction was quenched and the supernatants were analyzed by chromatography/mass spectrometry. Results In this study, nomilin and obacunone showed potent inhibitory effect on CYP3A4 with the IC50 values of 3.50 and 6.08 μmol/L, respectively. The inhibition of CYP3A4 was in a time-, concentration- and NADPH-dependent manner with Ki values of 2.92 and 1.25 μmol/L and Kinact values of 0.033 and 0.078 min-1 for nomilin and obacunone respectively. These results elucidated that they were time-dependent inhibitors for CYP3A4. Conclusion Concomitant use of limonoids and other drugs may call for extra caution for purposes of clinical safety.

Decomposition and Reformulation of Banxia Baizhu Tianma Decoction: A Vasodilatory Approach

2017/5/2 17:04:13

Objective To decompose and reformulate Banxia Baizhu Tianma Decoction (BBT) in order to optimize its ratio for vasodilatory effect. The fingerprints of these six component herbs were also studied via Tri-step FTIR analysis. Methods The herbal components of BBT were extracted separately with different solvents, such as 95% ethanol, 50% ethanol, and distilled water. The vasodilatory effect of the extracts was studied using the in vitro pre-contracted aortic ring model. Among these extracts, the water extracts of Pinellia ternata [PT(W)] and Citri reticulatae [CR(W)], 50% ethanol extracts of Gastrodia elata [GE(50)] and Glycyrrhiza uralensis [GU(50)], and 95% ethanol extracts of Atractylodes macrocephala [AM(E)], and Poria cocos [PC(E)] were selected for the orthogonal stimulus-response compatibility group study due to their high effectiveness in vasodilation. Results The best combination ratio (G2) was GE(50), GU(50), AM(E), PC(E), CR(W), and PT(W) at EC0, EC5, EC5, EC5, EC5, and EC5, with EC50 of (0.06 ± 0.01) mg/mL. Comparison between the vasodilatory effects of extracts derived from G2 and BBT were also studied. From the results, the 50% ethanol extract following G2 ratio was found to be most potent extract with the highest vasodilatory effect compared with other extracts derived from G2 or BBT, with EC50 of (0.05 ± 0.005) mg/mL. Conclusion The optimum ratio and appropriate extraction method for the new BBT formulae are obtained. G2 is determined as the best formulae with the highest potency in producing the vasodilatory effect on isolated rat aortic rings. The results obtained suggest that 50% ethanol is found to be the best solvent to extract the vasoactive compounds of G2 in order to exhibit the highest efficacy of vasodilatory effect.

UPLC-Q-TOF/MS-based Metabolic Profiles of Bioactive Components in Rehmannia glutinosa and Cornus officinalis Herb Pair by Rat Intestinal Bacteria

Jin-hua Tao1,Min Zhao2,Yong Ling1,Shu Jiang2 and Wen-qian Qiu1 — 2017/5/2 17:04:14

Objective To investigate the metabolic routes and metabolites of Rehmannia glutinosa and Cornus officinalis herb pair produced by gut microbiome from rats. Methods A rapid and sensitive ultra-performance liquid chromatography/quadrupole-time-of- flight mass spectrometry (UPLC-Q-TOF/MS) technique combined with MetabolynxTM software was established and successfully applied to identify the metabolites of the main bioactive components in the herb pair extract by rat intestinal bacteria. Results Four parent compounds (loganin, morroniside, catalpol, and acteoside) and their eight corresponding metabolites were detected and tentatively identified by the characteristics of their protonated ions. Hydrogenated and demethylated loganetin, dehydroxylated morronisid aglycone, caffeic acid, and its methylated product were the main metabolites. These metabolites suggested that the glycosides were firstly hydrolyzed to their aglycones by hydrolytic enzymes of the enteric microbial flora and subsequently to the other metabolites through hydrogenation, (de)-methylation, and de-hydroxylation. Conclusion The results may be helpful for the further investigation of the pharmacokinetic study of R. glutinosa and C. officinalis herb pair in vivo.

Identification of WRKY Transcription Factors Related to Saikosaponin Biosynthesis in Adventitious Roots of Bupleurum chinense

Su-rui Wu,Ke Gao,Xuan Liu,Jiao Xu,Jian-he Wei and Chun Sui — 2017/5/2 17:04:14

Objective To identify the genes of WRKY transcription factors (TFs) from roots of Bupleurum chinense and genes that potentially regulate saikosaponin (SS) biosynthesis. Methods Firstly, the subfamily cluster analysis was mainly based on Arabidopsis thaliana WRKYs for 27 putative WRKY TFs selected from previous transcriptome sequencing data. Secondly, qPCR was used to screen such genes of WRKY TFs that could be induced by NaCl and PEG6000 in adventitious roots of B. chinense. Meanwhile, saikosaponins (SSs) in treated adventitious roots were determined by HPLC. The roots were collected at 0, 2, 4, 8, 12, 24, 48, and 72 h after treatments, and 120 h only for PEG. Finally, the tissue-specific expression was analyzed on screened genes by qPCR. Results The 27 genes were grouped into three categories: There were nine in Group I, 15 in Group II, and two in Group III. Four genes of WRKYTFs, BCWRKY6, BCWRKY16, BCWRKY32, and BCWRKY35 were obviously induced by NaCl in adventitious roots of B. chinense, while only BCWRKY32 was induced by PEG. The content of SSs increased at different levels in NaCl and PEG6000 treatment. Three genes including BCWRKY6, BCWRKY32, and BCWRKY35, expressed most in roots, were similar to the accumulation pattern of SS. Conclusion The three WRKY genes, BCWRKY6, BCWRKY32, and BCWRKY35, may be involved in the biosynthesis of SS.

Efficacy of Traditional Chinese Medicines in Preventing Oxaliplatin-induced Peripheral Neurotoxicity in Cancer Patients: A Network Meta-analysis

Xiao-chen Wei1,Li-qin Zhu1,Hui Wang2,Chun-ge Wang1,Qi Deng1 and Xin Li1 — 2017/5/2 17:04:14

Objective Oxaliplatin-induced peripheral neurotoxicity (OIPN) is the main limitation for its continuation in cancer patients. Traditional Chinese medicines (TCMs) have been used to prevent OIPN in China and have been demonstrated to be effective. However, due to the lack of direct comparison among TCMs, it remains unclear that which TCM is the best for OIPN prevention. Consequently, the present study aimed to compare the relative efficacies of TCMs to find out the best TCM by applying a network meta-analysis. Methods Studies were identified by searching PubMed, EMbase, Cochrane Libraries, CNKI, WanFang, and WeiPu database from January 1990 to May 2016. Randomized controlled trials (RCTs) that evaluated the efficacy of TCMs in preventing OIPN in cancer patients were included. Statistical analysis was performed with ADDIS 1.16.6. Results Twenty-five RCTs (1572 patients) involving five TCMs were included. The results of network meta-analyses showed that compared with oxaliplatin-based chemotherapy alone, the combination with Huangqi Injection (HQI), Shenmai Injection (SMI), Shenfu Injection (SFI), Buyang Huanwu Decoction (BHD), and Huangqi Guizhi Wuwu Decoction (HGWD) could decrease the overall OIPN incidence and the severe OIPN incidence in cancer patients. In addition, probability ranking results showed the order of efficacy in preventing overall OIPN incidence was HQI > HGWD > SFI = SMI > BHD, while the order of efficacy in preventing severe OIPN incidence was HQI > HGWD > BHD > SFI = SMI. Conclusion All five TCMs are effective neuroprotective agents against OIPN. Among these TCMs, HQI and HGWD were superior to others in clinical efficacy. Moreover, Astragalus membranaceus might be a more promising herb for the OIPN prevention. However, more direct head-to-head RCTs with high quality and large sample size are still needed to further confirm the conclusion.

Screening of Eleven Commonly Used Traditional Chinese Medicines for Inhibitory Effects on Human Cytochrome P450 Enzymes

2017/5/2 17:04:14

Objective Traditional Chinese medicines (TCMs) as natural therapeutic agents have been widely used and received great attention over the world. However, the information on them to cause herb-drug interactions mediated by cytochrome P450 (CYP) enzymes is limited. The present study aims to evaluate the inhibitory effects of 11 commonly used TCMs, including Nelumbinis Folium, Ginseng Radix et Rhizoma, Ziziphi Spinosae Semen, Chrysanthemi Flos, Lonicera Japonica Flos, Lycii Fructus, Cassiae Semen, Poria, Crataegi Fructus, Schisandrae Chinensis Fructus, and Polygonati Rhizoma, on the five human major CYP isoenzymes 2C19, 2D6, 3A4, 2E1, and 2C9. Methods An in vitro cocktail method was used. Results The aqueous extracts of all tested TCMs were found no significant inhibitory effect, with IC50 values higher than 100 μg/mL. While, the ethanolic extracts of some herbs showed more potent inhibition. These herbs include Nelumbinis Folium with IC50 values of 12.05 and 61.43 μg/mL on CYP2D6 and CYP2E1, Schisandrae Chinensis Fructus with IC50 values of 64.42 and 47.24 μg/mL on CYP2C19 and CYP3A4, and Chrysanthemi Flos with IC50 values of 45.25 μg/mL on CYP2C9, respectively. Conclusion Co-administration of these TCMs and their products with conventional medicines should be paid much attention to, and their potential inhibitory effects on CYP enzyme activities need to be further investigated.

Anti-inflammatory Effects of Saxifrage Cream in Hapten-induced Chronic Dermatitis-eczema in Mice

Jun Yang,Yong-Ai Xiong,Shi-Ji Xiao and Ying-Shu Xu — 2017/5/2 17:04:14

Objective Chronic dermatitis-eczema is a kind of inflammatory skin disease characterized by pruritic and eczematous skin lesions. In the present study, we investigated the anti-allergic effect of Saxifrage cream on eczema-like skin lesions induced by 2,4-dinitrochlorobenzene (DNCB) in mice. Methods KM mice were randomly divided into eight groups: normal group (Norm), model group (DNCB), positive group 1 (Dexamethasone cream, DEX), positive group 2 (Chinese herbal medicine Paeonol cream, PA), Saxifrage cream groups (equivalent to crude herb 1.25, 2.50, 5.00 g/g, S1, S2, S4), and negative group (Norm + S4). Except for the normal control group and negative control group, right ears of mice in other groups were repeatedly induced with 0.5% DNCB for four times for 3 d and previously sensitized with 7% DNCB to induce eczema. Then mice in different groups were treated by DEX, PA, S1, S2, and S4, respectively. After mice were sacrificed, weight difference of two ears, immune organ change, and pathological change of right ear were used to assess the effects of the drugs. Results Compared with model group, DEX, PA, S2, and S4 could significantly lower mice ear thickness and swelling degree, and improve eczema skin symptoms. Meanwhile, the index of thymus and spleen in treatment groups were significantly reduced (P < 0.05). What’s more, the histological analysis demonstrated that thickening of the skin lesions were significantly reduced in DEX, S2, and S4-treated group, and the levels of IL-4 and IL-6 in serum could be reduced by DEX, S2, and S4. Conclusion Saxifrage cream has a protective function against DNCB-induced eczematous. The mechanism might be related to alleviate eczematous inflammatory reaction.

Inhibition of Yuyihe Powder on Tumor Growth in Mice Models of Sarcoma 180

2017/5/2 17:04:14

Objective To explore the antitumor effect of Yuyihe Powder (Yu Yi He San, YYHS) and its antitumor mechanism. Methods After treatment, tumor weight, immune apparatus weight, the life span of transplanted animals, spleen lymphocyte proliferation assays, and IL-2 concentration in mouse serum were recorded or detected. Results YYHS showed strong antitumor ability. Compared with control group, mid-dose YYHS (1.0 g/kg) could inhibit the tumor growth, prolong the life span of S180-bearing mice to some extent, significantly increase the thymic and splenic indices of S180 mice, and strongly promote the secretion of IL-2 in blood; The inhibitory rate on tumor growth and life prolongation rate were 37.1% and 38.37%, respectively. Conclusion YYHS could not only significantly inhibit the growth of S180 cells, but also markedly prolong the survival time of S180 bearing mice. The mechanism of antitumor effect could obviously enhance immunologic function of the S180 bearing mice to inhibit the growth of S180 cells.

Distribution of Metabolites in Root Barks of Seven Tree Peony Cultivars for Quality Assessment Using NMR-based Metabolomics

Pei Wang,Ze-ming Rong,Cui-xia Ma,Xin-feng Zhao,Chao-ni Xiao and Xiao-hui Zheng — 2017/1/23 9:58:31

Objective To determine the distribution of metabolites in the root barks of different tree peony cultivars for quality assessment. Methods Seven tree peony phenotypic cultivars with different colors were systematically analyzed using NMR-based metabolomics. Results A total of 16 metabolites from their methanol extracts were simultaneously identified and quantified, including one primary metabolite (sucrose) and 15 secondary ones (acetophenones, phenolics, monoterpene glycosides, flavonoids, and unsaturated fatty acids). The quantitative data indicated that sucrose (90-180 mg/g) and acetophenones (15-100 mg/g), and non-phenolics, monoterpene glycosides, flavonoids, and unsaturated fatty acids (2-15 mg/g) were the major metabolites in these tree peony cultivars. The significantly increasing levels of paeonoside with bioactivity were observed in “Xiangyu”, “Wujinyaohui”, “Roufurong”, “Yaohuang”, “Zhaofen”, “Doul?”, and “Yingrihong” in order. Opposite trends in the levels of paeonoside and paeonol were observed in “Xiangyu” and “Yingrihong”, suggesting that the changes of the secondary metabolites in plants were influenced by primary metabolites, such as sucrose/glucose, and the different physiological processes occurred in different tree peony cultivars. Conclusion “Yingrihong” with red flower has the highest medicine quality whereas “Xiangyu” with white flower has the worst one based on the content of paeonoside.

Simultaneous Determination of Seven Components in Gamboge and Its Processed Products Using a Single Reference Standard

2017/1/23 9:58:31

Objective To establish a quality control method for simultaneous determination of multiple components in gamboge. Methods A single reference standard for the determination of multiple components (SSDMC) with HPLC was proposed. Seven major components of gamboge including gambogenic acid (S), β-morellic acid (C1), 2R-30-hydroxygambogic acid (C2), isogambogenic acid (C3), gambogellic acid (C4), 2R-gambogic acid (C5), and 2S-gambogic acid (C6) were simultaneously analyzed using gambogenic acid as reference standard. The credibility and feasibility of SSDMC method were validated with respect to linearity, limits of detection and quantification, precision, stability, repeatability, accuracy, ruggedness, and robustness. The relative conversion factors (RCFs) of S and C1-6 were calculated. Twelve batches of gamboge including crude and processed products were successfully analyzed by applying the SSDMC and traditional external standard (ES) methods. Results The SSDMC method was credible and feasible. The RCFs of S and C1-6 were 1.000, 0.913, 0.864, 1.064, 0.777, 0.921, and 0.919, respectively. No significant difference was observed in the contents of the seven components between SSDMC and ES methods. The heat-processing technique caused a reduction in the seven components. Conclusion SSDMC is a simple, reliable, and effective method for the analysis of the complex multiple components in gamboge, and it is also a practical and economical approach.

Analysis on Isozyme and Diterpene Lactones Variability during Ontogenesis of Andrographis paniculata

Jun-li Gao,Yan-hua Shao,Xiao-ping Lai and Ping Ding — 2017/1/23 9:58:31

Objective To understand the relationship between isozyme activities and diterpene lactone biosynthesis of Andrographis paniculata. Methods Plants were collected during ontogeny from seeding to seed maturity, and separated into leaves and stems for determination. Morphological and yield parameters were used to describe plant growing states. Isozyme changes were tested by polyacrylamide gel electrophoresis. HPLC was used to develop the fingerprints as well as to determine the diterpene lactone content. Results Significant increases were observed in the activities of isozymes, such as aspartate aminotransferase (AST), malate dehydrogenase (MDH), peroxidase (POD), and catalase (CAT), around the early stage of bud in leaves, and the activities of these four kinds of isozymes increased gradually as time progressed in stems. The content changes of diterpene lactones in leaves and stem were various. In the leaves, andrographolide (1) was recorded the highest [(23.63 ± 1.06) mg/g] at the early stage of bud, whereas deoxyandrographolide (2) was the lowest [(6.78 ± 0.27) mg/g] at this period and it reached the highest level at the seeding stage [(26.05 ± 1.04) mg/g]. Dehydroandrographolide (3) and neoandrographolide (4) fluctuated during growing stages. Meanwhile, the HPLC fingerprint showed that the content changes of two unknown compounds were related to that of dehydroandrographolide in leaves. In stems, andrographolide had increased gradually until the bud stage [(8.26 ± 0.33) mg/g], and other three diterpene lactones showed a trend of fluctuation. The yield of total diterpene lactones in aerial part reached the highest at the first flowering stage (806.71 mg/plant). Conclusion These results lay the foundation for the future research on the relationship of isozymes and diterpene lactones, and for determining the most favorable time for harvesting A. paniculata.

2-(2-Phenylethyl)chromones from Endophytic Fungal Strain Botryosphaeria rhodina A13 from Aquilaria sinensis

2017/1/23 9:58:31

Objective To study the characteristic 2-(2-phenylethyl)chromone components of endophytic fungal strain of Aquilaria sinensis by solid culture. Methods The compounds were isolated by various chromatographic methods such as silica gel, reverse-phase silica gel, Sephadex-LH20 column chromatography as well as crystallization. Results Seven 2-(2-phenylethyl)chromone analogues were isolated from the solid culture of Botryosphaeria rhodina A13. Their structures were established by spectral data as well as physicochemical properties, and identified as 6-hydroxy-7-methoxy-2-(2-phenylethyl)chromone (1), 6,7-dimethoxy-2-(2-phenylethyl) chromone (2), (5S,6R,7S,8R)-2-(2-phenylethyl)-5,6,7,8-tetrahydrchromone (3), 6- hydroxy-2-(2-phenylethyl)chromone (4), 4′-hydroxy-2-(2-phenylethyl)chromone (5), 6-methoxy-2-phenethyl-4H-chromen-4-one (6), and 6-methoxy-2-(4′-methoxy- phenethyl)-4H-chromen-4-one (7). Conclusion All of the compounds are isolated for the first time from the genus Botryosphaeria. This research opens up a new vista to produce the characteristic components of agarwood by endophytic fungi.

Antifungal Activity of Aspidin BB from Dryopteris fragrans against Trichophyton rubrum Involved Inhibition of Ergosterol Biosynthesis

2017/1/23 9:58:31

Objectives Aspidin BB, a typical phloroglucinol derivative from Dryopteris fragrans, possesses significant antifungal property. This study aimed to investigate potential mechanism of antifungal activity of Aspidin BB against Trichophyton rubrum which is the most common pathogens responsible for chronic dermatophytosis. Methods The minimum inhibitory concentration (MIC) of Aspidin BB against strains was determined by broth microdilution. The effects of Aspidin BB on ergosterol biosynthesis were investigated by content determination based on UPLC method. Besides, the effects of drugs on squalene epoxidase (SE) in T. rubrum cell membrane were analyzed. Results MIC value of Aspidin BB against T. rubrum was 25.0 μg/mL. Aspidin BB reduced ergosterol content significantly, but no notable effect on squalene epoxidase activity. Conclusion The results suggested that Aspidin BB inhibited ergosterol biosynthesis. However, it was not squalene epoxidase but other components may sever as possible targets in ergosterol biosynthesis pathway.

Bioactive Study on Mixtures of Epimedin C and Icariin Monomers with Invariant Molarity to Zebrafish Osteoporosis Model

2017/1/23 9:58:31

Objective To evaluate the anti-osteoporosis effect of the mixtures of epimedin C and icariin monomers with invariant molarity on zebrafish osteoporosis model. Methods The zebrafishes, fertilized 4 d ago, were exposed in 11 groups of nutrient solutions with prednisolone (25 μmol/L) as well as epimedin C and icariin (15 μmol/L) of various contents. The ratio of epimedin C and icariin in the 11groups were as follows: A (10:0), B (9:1), C (8:2), D (7:3), E (6:4), F (5:5), G (4:6), H (3:7), I (2:8), J (1:9), and K (0:10). Meanwhile, a negative control group with prednisolone (25 μmol/L) was prepare as S. The selected zebrafish fetus was put into the 24-hole culture plate, and ensure every 5 zebrafishes in 1 hole and 2 holes as a group. They were placed in incubator at 28.5 oC, and the daily changes of fluid were investigated until they were put to death on day 8 and then fixed. After dyeing with alizarin red, the segmental venter of zebrafish skulls was observed and quantitative analysis of dyed area was conducted. Results Compared with the negative control group S, the integrated optical density (IOD) values of cranial dyed area in all groups increased significantly (P < 0.05); Compared with group S, the IOD value of cranial dyed area in mixtures of epimedium monomers increased significantly (P < 0.05). The mixtures of epimedium monomers were all effective in facilitating zebrafish cranial mineralization and preventing prednisolone from inducing osteoporosis. According to mixtures of A-K groups, zebrafish cranial mineralization gradually decreased with gradually reduced content of epimedin C, with significant difference among groups (P < 0.05). Conclusion The higher the content of epimedin C in mixtures with invariant molarity is, the more active the anti-osteoporosis effect of epimedinC to zebrafish osteoporosis model is.

Developmental Toxicity and Potential Teratogenicity of Compound Danshen Tablet, Angong Niuhuang Pill, and Lidan Paishi Tablet in Zebrafish Embryos

2017/1/23 9:58:31

Objective Herbal medicines containing toxic herbs or minerals such as Compound Danshen Tablet (CDT), Angong Niuhuang Pill (ANP), and Lidan Paishi Tablet (LPT) are avoided or used with caution for pregnant women because of potential teratogenicity. To understand their mechanism, they were chosen as model subjects for the research. Methods Zebrafish embryos were used to evaluate their potential teratogenic risk in vitro. Results All of them showed teratogenic and lethal effects in zebrafish embryos, with the EC50 values at 351, 793, and 220 μg/mL, and LC50 values at 417, 596, and 380 μg/mL, respectively. CDT and LPT, displaying week potential teratogenicity as their teratogenicity indexes were greater than 1, induced tail malformation and cardiac edema mainly in zebrafish embryos, respectively. Conclusion The results provide the significant guidance of clinical safety of medication.

Anti-flu Effect of Compound Yizhihao Granule and Its Effective Components

Hua Huang,Hua Yao,Lin-lin Wang,Li-jun Si,Qiao-li Yang and Zheng-yi Gu — 2017/1/23 9:58:31

Objective To research the anti-influenza effect of active ingredients in Compound Yizhihao Granule (CYG). Methods The cytotoxicity and cytopathic effects (CPE) were observed under the phase-contrast microscope, besides 50% toxicity concentration (TC50) and 50% inhibitory concentration (IC50) were also calculated using Reed-Muench method, then the antiviral activity in vitro according to Selection Index (SI = TC50/IC50 ) was evaluated. In PR8 virus-infected mice, survival time, death rate, and lung index were observed in order to evaluate the protective effect. Besides, the effective ingredients were determined using HPLC method, and their contents were calculated by external standard method. Results CYG could inhibit the influenza virus-induced CPE, with IC50 of 4.6 mg/mL (equal to herbal extracts 262.2 μg/mL), and no direct cytotoxic effect at this concentration. PR8-infected mice were ig given CYG, the lung index and mortality were significantly reduced, and survival time was obviously prolonged. HPLC analysis indicated CYG contained many kinds of antivirus active components, including rupestonic acid, epigoitrin, and adenosine. Conclusion CYG is an effective natural anti-influenza medicine. Its antiviral effect should be the synergic effect of a variety of antiviral active ingredients.

Pharmacognostical study of Cynanchum stauntonii and Cynanchum glaucescens, Botanical Sources of TCM Baiqian

2018/10/8 0:00:00

Objective Cynanchum stauntonii and Cynanchum glaucescens are botanical species of Baiqian (Cynanchi Stauntonii Rhizoma et Radix) in Chinese Pharmacopoeia, in which, however, there are no microscopic identification. So we provided the morphological and microscopic identification of the crude drug for updating Chinese Pharmacopoeia. Methods Twelve batches of C. stauntonii and three batches of C. glaucescens and their crude drugs were taxonomically, morphologically, and microscopically examined. Results Taxonomically, C. stauntonii had narrowly lanceolate leaves with acuminate apex and 5 mm long petiole; Whereas C. glaucescens was oblong-lanceolate or oblong with rounded or acute apex in leaves, and had very short or no petiole. Morphologically, rhizomes of C. stauntonii and C. glaucescens both had hollow pith, but the hollow pith occupied about a half of the rhizome’s diameter in C. stauntonii, whereas only a very small proportion of the overall diameter in C. glaucescens. Moreover, microscopic observation showed the difference in the proportion of xylem and in rhizome transverse-sections of the two species along with the difference in the size of the pith. Finally, laticifers and rhizome epidermal secretory cells were present in the powders of C. stauntonii, but absent from C. glaucescens. Conclusion Based on observation of morphological and microscopic characteristics, the two species can be distinguished by the size of the pith, proportion of xylem of rhizomes, and crude drug powder characters such as laticifers and secretory cells.

Quality assessment of crude and processed Leonuri Fructus by chemical and color analysis combined with chemometric method

2018/10/8 0:00:00

Objective: Crude Leonuri Fructus (CLF), the fruits of the Leonurus japonicus, and processed Leonuri Fructus (PLF) by stir-baking as the important Chinese herbal medicines, have been used in China and other Asian countries for thousands of years. The objective of this research is to reveal the difference between CLF and PLF. Methods: The sensory technologies of the colorimetry, sensitive and validated HPLC-ELSD and GC combined with flame ionization detector (GC-FID) were employed to discriminate CLF and its processed product PLF. The color parameters of the samples were determined by colorimetric instrument CR-410. Moreover, the content of stachydrine and six fatty acids were determined by HPLC and GC. Subsequently, analysis of variance (ANOVA), principal components analysis (PCA), hierarchical cluster analysis (HCA), and Kendall’s correlation test were performed for data analysis. Results: The CLF and PLF were divided into two categories by PCA and HCA in terms of their component content and color. The results distinctly demonstrated significant changes in color and the content of indicative components between CLF and PLF. Conclusion: The study revealed that HPLC, GC, and colorimetric method in combination with chemometric method could be used as comprehensive quality evaluation for CLF and PLF.

Multivariate modeling to evaluate effect of fabricating process on product profile of high shear granulation

Zheng Yan, Ying-ying Gong, Liang-shan Ming and Yi-hui Xie — 2018/10/8 0:00:00

Objective: This study aims to investigate the feasibility of high shear granulation (HSG) for agglomeration of Crataegi Folium extract (Chinese edible herbal), and explore the effect of process variables on granule critical quality attributes (CQAs) by three different models, i.e., response surface methodology (RSM), multilayer perceptron neural networks (MLP), and partial least squares method (PLS). Method: RSM, MLP, and PLS complementary to design of experiment (DoE) were utilized to investigate the influence of granulation time, impeller speed, and binder amount on the products. Crataegi Folium extract was employed as granulation powder. RSM was further carried out to develop design space of HSG. Results: The results indicated that RSM, MLP, and PLS modeling techniques enhanced the understanding and controlling of granules produced via HSG. The granule CQAs were mainly influenced by granulation time, impeller speed, and binder amount. Overlay plots of the RSM indicated that design space for the operating ranges of impeller speed and binder amount at high levels of granulation time was the smallest. Conclusion: The study showed that these models were useful to characterize the granulation process, and was particularly important to understand the process.

Mechanism of Chinese herbal formula QHF against breast cancer MCF-7 cells invasion and migration

2018/10/8 0:00:00

Objective: To investigate the effects of Chinese herbal formula Qinghuofu (QHF) on the migration and invasion of breast cancer MCF-7 cells and its possible molecular mechanisms, thereby providing a theoretical basis to find effective anti-cancer medicine and therapeutic targets for the treatment of anti-migration and anti-invasion of breast cancer. Methods: Breast cancer MCF-7 cells were treated with different QHF and other different reagents, CCK8 assay was used to detect the influence of the reagents on the proliferation of MCF-7 cells; Scrape migration and Transwell assay were used to quantitatively determine the migration and invasion effects of QHF and HGF on the MCF-7 cells. Subsequently, the c-Met inhibitor and its downstream ERK and PI3K inhibitors were used to investigate the relationship between the migration and invasion of MCF-7 cells, as well as its downstream MAPK/ERK and PI3K/Akt signaling pathways. The expression levels of HGF, c-Met, ERK, p-Akt, p-c-Met, p-ERK, p-Akt, MMP2, MMP9, and VEGF in breast cancer MCF-7 cells treated with QHF and other reagents were also examined. Results: The result indicated that formula QHF not only significantly inhibited the proliferation of MCF-7 cells, but also significantly suppressed the effects of (hepatocyte growth factor) HGF (40 ng/mL) on the proliferation and movement of MCF-7 cells, reducing the ability of the cells to invade and migrate. Western blot analysis indicated that QHF and c-Met inhibitor significantly decreased the expression of p-c-Met, p-ERK1, p-ERK2, p-Akt, MMP-2, MMP-9, and VEGF, while HGF could significantly increase the expression of p-c-Met in MCF-7 cells; c-Met downstream ERK and PI3K inhibitors could also significantly decreased the expression of MMP-2, MMP-9, and VEGF in MCF-7 cells; But the difference among c-Met, PI3K, ERK, and QHF group were not statistically significant. Conclusion: QHF can prevent the proliferation, migration, and invasion of MCF-7 cells by inhibiting the HGF/c-Met and its downstream PI3K/Akt and MAPK/ERK signaling pathways; Thereby down-regulating the expresssion of HGF, p-Met, p-ERK1, p-ERK2, p-Akt, MMP-2, MMP-9, and VEGF.

Antifibrosis effects of Shenge Yangfei Capsules on bleomycin-induced pulmonary fibrosis in rats

2018/10/8 0:00:00

Objective: The present study, for the first time, was designed to examine the antifibrosis effects of Shenge Yangfei Capsules (SGYF) on bleomycin-induced pulmonary injury and fibrosis in rats. Methods: Animals were divided into six groups randomly: saline control group, bleomycin group, and SGYC (70, 420 or 850 mg/kg/d, ig) group, and positive control group. Animals were sacrificed on day of 7th, 14th, and 28th post bleomycin administration, and lungs were removed respectively. For histological evaluation of lungs injury and fibrosis, lung specimens were stained with hematoxylin and eosin (HE) and Masson trichrome. Body weight and lung index from various groups were measured, and the concentrations of monoamine (MAO) and transforming growth factor-β1(TGF-β1) were also detected in lung homogenates. Results: SGYC attenuated the bleomycin-induced weight loss and the lung index. Histologica levidence showed the ability of SGYC to decrease bleomycin-induced lung fibrosis and consolidation. SGYC reduced the MAO and TGF-β1 activityin lung tissues. Conclusions: These findings suggest that SGYC may be a promising candidate to prevent bleomycin-induced lung damage or other interstitial lesions.

Optimization of enzyme assisted extraction of polysaccharides from pomegranate peel by response surface methodology and their antioxidant potential

2018/10/8 0:00:00

Objective To optimize the enzymatic extraction conditions of polysaccharides from pomegranate peel using response surface methodology (RSM), and invest the anti-oxidant activity of pomegranate peel polysaccharide in vitro for seeking novel biological components used as pharmaceutical products and functional foods. Methods Effects of enzymolysis time, ratio of water to raw material, and the dosage of cellulase on the extraction yield of pomegranate peel polysaccharide were fully examined by Box-Behnken design of RSM. DPPH radical scavenging activity, hydroxyl radicals scavenging activity, superoxide anion radical scavenging activity, and reducing power assay were measured by microplate reader. Results The optimal extraction conditions were as follows: enzymolysis temperature 55 ?C, pH 5.0, enzymolysis time 88 min, ratio of water to raw material 22:1 mL/g, and dosage of cellulose 0.93%. Under the optimal extraction conditions, the yield of pomegranate peel polysaccharide was (22.31 ± 0.07)%, which was well matched with the predicted value 22.35% of the Box-Behnken design model. Also, pomegranate peel polysaccharide demonstrated appreciable anti-oxidant potential on DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging and reducing powerin vitro. Conclusions An optimized enzymolysis assisted method was proposed for the extraction of pomegranate peel polysaccharide, which can be used as a good anti-oxidant.

Intervention effects of Compound Houttuyniae Herba to diabetic renal damage based on SOCS-JAK/STAT negative feedback regulation

Yun Fang, Sai-cong Shao and Hai-ying Wang — 2018/10/8 0:00:00

Objective To research the protective effects of different extracts from Compound Houttuyniae Herba (CHH) and its mechanism through JAK/STAT-SOCS-1 signaling pathway. Methods The normal group comprised db/m mice (n = 8). db/db mice were randomly divided into seven groups with eight mice in each group according to the applied treatment method: model group, metformin hydrochloride (MH) group, AG490 group, water extract (WE) group, ethanol extract (EE) group, volatile oil (VO) group, and mixture (MG) group (mixture of above three extract) of CHH. After 8 weeks, the general status, biochemical indicators, and renal histological changes in the mice were evaluated, the total RNA and protein were collected and RT-PCR method was used to examine the effect on mRNA expression of JAK2, STAT3, SOCS1, and Western blot method was used to detect the protein expression of JAK2, P-JAK2, STAT3, P-STAT3, SOCS1, c-fos, and c-jun. Immunofluorescence was used to observe the protein expression of c-fos and c-jun in kidney tissue. Results Compared with normal group, the serum level of TGF-β1, FN of EE, VO, and MG groups were decreased (P < 0.05). Renal function was also improved but not significantly. Also, the renal histology was improved especially in the mixture group. The protein expression of JAK2, STAT3, P-JAK2, P-STAT3, and the genetic expression of JAK2 and STAT3 in kidney tissue were significantly increased in the model group (P < 0.05). Compared with the model group, the expression of P-JAK2 and P-STAT3 was down-regulated in treatment groups; The expression of SOCS-1 of VO and mixture groups were elevated (P < 0.05). Conclusion CHH has beneficial effects on diabetic renal injury, which may protect and improve the kidney function and reduce urinary protein, maintain the integrity of the kidney structure and function.

Potential mechanism of cholagogic effect of Gardenia jasminoides Zhizi: Reduction of hepatic bile acids level by increasing urinary excretion via up-regulating renal Mrp2 in normal rats

2018/10/8 0:00:00

Objectives Gardenia Jasminoides (Zhizi in Chinese), belonging to Rubiaceae family, has been traditionally used to treat cholestasis and jaundice for centuries in Asia. In the theory of traditional Chinese medicines, Zhizi could dispel dampness and heat via the urine to execute its choleretic effects. However, the potential molecular mechanism is still poorly clarified. Here, we investigated the effect of different dose of Zhizi aqueous extracts powder on urinary excretion of bile acids (BAs), and explored the potential mechanism via renal BAs efflux transporters Mrp2 and Mrp4 in normal rats. Methods Male Wistar rats were orally administrated with 0.3 or 0.9 g/kg/d dose of Zhizi aqueous extract powder for 2 weeks, then body weight, serum aminotransferase, total BAs concentrations in liver, bile, serum, kidney, and urine, 1 h bile flow, 12-h urinary volume, biliary, and urinary excretion amount of total BAs as well as protein expression of major renal BAs efflux transporter Mrp2 and Mrp4, were all evaluated. Results Zhizi especially the high dose of Zhizi aqueous extract powder reduced hepatic total BAs concentration. Additionally, bile flow and biliary excretion had no significant difference, but the remarkable increased urinary excretion of BAs and 2 ? 3 folds up-regulated renal Mrp2 expression were observed after administrated with Zhizi as compared with the control group. Conclusion The findings indicated that Zhizi reduced hepatic total BAs level by increasing urinary excretion rather than the biliary excretion of BAs, which in turn ascribed to elevated protein expression of Mrp2 at apical membrane of renal tubular epithelial cells.

Xanthotoxin induces apoptosis in SGC-7901 cells through death receptor pathway

Xiu-juan Zhang, Cang Wei, Li-juan He and Jian An — 2018/10/8 0:00:00

Objective To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro. Methods SGC-7901, HepG-2, MCF-7, and A549 cells were treated with different concentrations of xanthotoxin (10, 20, 60, 80, 100, 120, 140, and 160 μg/mL) for 48 h, and the cell viability (IC50) was determined by MTT assay; Xanthotoxin-induced apoptosis in cells was observed by using Hoechst 33258 Staining Kit and Annexin V-FITC Apoptosis Detection Kit; Flow cytometry was used to detect apoptosis related proteins of Fas/FasL, Bid, and DR5/TRAIL proteins in human gastric carcinoma SGC-7901 cells after being treated by xanthotoxin; The influence of xanthotoxin on Caspase-8 protein expression in the cells was determined by Flouormetric Assay Kit. Results Xanthotoxin obviously inhibited SGC-7901, HepG-2, MCF-7, and A549 cells proliferation, and its inhibition was in a concentration-dependened manner; Flow cytometry results showed that in a certain concentration range, xanthotoxin can increase the expression levels of Fas/FasL and DR5/TRAIL proteins in a concentration-dependence manner. The content of bid protein in cells was increased, and it showed concentration-dependence. Conclusion Xanthotoxin may induce SGC-7901 cells apoptosis in a certain concentration range through the Fas/FasL protein mediated death receptor pathway, or by DR5/TRAIL mediated death receptor pathway, and increase the expression level of death receptor protein, activation Caspase-8, activating downstream effect factor, inducing cell apoptosis, or activate Caspase-8 cutting activate protein Bid, and then enter the mitochondrial pathway, induction of apoptosis.

Effect of cinnamaldehyde on 1, 3-β-d-glucans in cell wall of Candida albicans in immunosuppressed BALB/c mice

Jie-hua Deng, Gang-sheng Wang, Ji-hong Li and Xiao-lu Wang — 2018/10/8 0:00:00

Objective: Invasive pulmonary candidiasis is a disease with high incidence, difficult treatment, poor prognosis, and high mortality. The present study analyzed the influence of cinnamaldehyde on 1,3-β-D-glucans in the cell wall of Candida albicans in order to provide a theoretical basis for the research of antifungal drugs. Methods: An immunosuppressed BALB/c mouse model with invasive pulmonary candidiasis was established by nasal perfusion of 50 μL of C. albicans suspension (107 cfu/mL). 1,3-β-D-glucans examination and electron microscopy were carried out. Fluconazole was used as the control. Results: Cinnamaldehyde was administered at a dose of 240 mg/kg/d for 14 consecutive days, and the measured value of 1,3-β-D-glucans was (1160.62 ± 89.65) pg/mL, whereas that of fluconazole was (4285.87 ± 215.62) pg/mL. The difference between the two groups was statistically significant (P < 0.05). Electron microscopy observation indicated that the 2?3 layers outside the cell wall of C. albicans (1,3-β-D-glucans layer) were rough, deformed, and incomplete, although the cell membrane was clear and intact. Conclusion: Cinnamaldehyde demonstrated special efficacy on 1,3-β-D-glucans in the cell wall of C. albicans.

Mechanism underlying bergapten-mediated regulation of vincristine transport in MDCK-MDR1 cells

2018/8/3 0:00:00

Objectives To investigate the effects of bergapten of Angelicae Dahuricae Radix on the transport of vincristine and its possible mechanism. Methods The apparent permeability coefficient (Papp) for the transport of vincristine through the membrane of MDCK-MDR1 cells was used as an indicator of the effect of bergapten on vincristine transport. Molecular docking was employed to predict the binding force between bergapten and P-glycoprotein (P-gp). The effects of bergapten on P-gp function and P-gp ATPase activity were determined by rhodamine 123 (Rho123) accumulation and activity analysis, respectively. 1,6-Diphenyl-1,3,5-hexatriene (DPH) was used to study the effects of bergapten on membrane fluidity, and Western blotting and quantitative real-time PCR assays were performed to analyze the effect of bergapten on the protein and mRNA expression of P-gp, respectively. These experiments clarified the effects of bergapten on the transport of vincristine and allowed exploration of the possible mechanism underlying the effects of bergapten. Results The results showed that bergapten could inhibit the transport of vincristine in MDCK-MDR1 cells, and the binding force between bergapten and P-gp was weaker. Bergapten could reduce the accumulation of Rh123 in MDCK-MDR1 cells, increase the membrane fluidity, and upregulate P-gp protein and mRNA expression but it had no effect on P-gp ATPase activity. Conclusions Overall, we concluded that the possible mechanism through which bergapten inhibits vincristine transport was related to the bergapten-mediated upregulation of P-gp protein and mRNA expression, membrane fluidity or P-gp enzyme activity.

Transcriptional activity and subcellular location of SmWRKY42-like and its response to gibberellin and ethylene treatments in Salvia miltiorrhiza hairy roots

Zhen-qing Baia,b, Wen-rui Lid,e, Zi-yun Zhoua and Zong-suo Lianga,c — 2018/8/3 0:00:00

Objective: The exogenous gibberellin (GA) and ethylene (ET) treatment can improve the medicinal ingredients of Salvia miltiorrhiza. Interestingly, many reports pointed out that WRKY transcription factors played an important regulatory role in these treatment responses. However, whether the SmWRKY mediate these treatment signalings in S. miltiorrhiza remains largely elusive. Methods qRT-PCR was used for SmWRKY42-like in response to exogenous GA and ethephon (Eth) treatment. The subcellular location of SmWRKY42-like was transiently transformed into onion epidermal cells by particle bombardment. The self-activating activity of SmWRKY42-like was verified in AH109 yeast strain. Results: SmWRKY42-like was a WRKY family gene in S. miltiorrhiza. The subcellular localization and transcriptional activity results of the SmWRKY42-like protein indicated that SmWRKY42-like mainly enriched in nucleus and might be a transcription factor in S. miltiorrhiza. In the meantime, the SmWRKY42-like gene significantly responded to exogenous GA and Eth treatment. Conclusion: These results collectively indicated the SmWRKY42-like gene functions, as an important hormone-responsive gene, might play a potentially role in ET and GA signaling pathways.

Medicinal plants of Chinese Pharmacopoeia and Daodi: insights from phylogeny and biogeography

Di Lei1, Jie Wu2, Christine Leon3, Lin-fang Huang 4 and Julie A. Hawkins1 — 2018/8/3 0:00:00

Objective The Chinese Pharmacopoeia (2015) includes 584 plant medicines, of which 284 also contain high quality subsets, so called “Daodi” components, where Daodi denotes superior clinical properties compared to non-Daodi counterparts despite being sourced from the same species. Commercial and clinical drivers of selection for Daodi have been described elsewhere. Our objective is to investigate the overall composition of Daodi to determine in what ways medicines with Daodi as a whole differ from the other plants of the Chinese Pharmacopoeia. A further objective is to characterise the Chinese Pharmacopoeia and Daodi in terms of the plant species including their traits and distribution.Methods We used trait analysis to identify whether Daodi species were significantly different from the remaining Chinese Pharmacopoeia plant species in any traits. We used biogeographic methods and an existing classification of Daodi into 10 regions to identify spatial patterns amongst the species. Regression and binomial analyses were used to test for over- and under-use of plant families and endemic species. Preferences for lineages were visualized using phylogenetic mapping. Results We found Daodi species (species with any Daodi subset) were more likely to be roots that are ‘hot’ or ‘warm’, and less likely to be ‘toxic’, according to traditional Chinese medicine (TCM) concepts. Roots were over-represented in the Bei region, and whole plants over-represented in Guang. Both the Chinese Pharmacopoeia and Daodi indicated preferences for families not common in previously studied ethnopharmacopoeias, and fewer endemic species were represented than expected by chance.Conclusion Using the phylogenetic and biogeographical methods, we highlighted patterns of plant use, and the biological characters of Daodi medicinal plants. Our study points towards cultural preferences in need of scientific explanation.

Antifatigue effects of peptide isolated from sheep placenta

2018/8/3 0:00:00

Objective: Fatigue has become one of the major threats to human health in the 21st century. Traditional Chinese medicine (TCM), which proved to be safer and more effective, has become a hot spot in antifatigue research. Human placenta, also called “Ziheche”, has drawn great attention in the antifatigue effect since the Tang dynasty. However, the shortage of human placenta restricts wide usage of it. According to the theory of TCM, sheep placenta (SP) also has the effect of nourishing blood, tranquilization, nourishing skin, and prolongation of life. The aim of this study was to examine the antifatigue effects of sheep placenta peptide (SPP), an extract of sheep placenta, in mice and the mechanism was also studied. Methods: Peptide from fresh SP was extracted via enzymolysis. SPP (0.13 g/kg) and soybean peptide (0.65g/kg) were administrated orally and daily to mice for four weeks. Antifatigue effects of SPP were estimated on weight-loaded swimming test; A non-weight-loaded swimming test was conducted to elucidate underlying the mechanisms of the anti-fatigue effects. Results: Administration of SPP prolonged the weight-loaded swimming time in mice. In addition, SPP significantly decreased the levels of muscle malondialdehyde (MDA) and serum lactic acid (LD), and increased the activities of muscle glutathione peroxidase (GSH), and superoxide dismutase (SOD) and liver glycogen in mice after non-weight-loaded swimming test. Moreover, the electron microscope observation showed that the muscle fiber bundle ranked neatly, the H band, I band, Z line as well as M line were clear and the numbers of mitochondria was normal though some of the mitochondria were swell in SPP treated mice after non-weight-loaded swimming test. Conclusion: SPP possesses potent abilities for antifatigue; Increasing the anti-oxidant activities and energy reserve as well as improving the ultrastructures in gastrocnemius muscle cells, which may be the mechanisms of SPP exerting its antifatigue effects.

Polyclonal antibody preparation of BcbZIP134 for transcriptional regulation analysis on saikosaponin biosynthesis

Jiao Xu, Xuan Liu, Su-rui Wu, Chun Sui and Jian-He Wei — 2018/8/3 0:00:00

Objective The aim of the present study was to verify how a transcription factor BcbZIP134 regulates the synthesis of saikosaponin using specific antibody. However, it is hard to obtain this soluble protein expressed in vitro, a prerequisite for antibody preparation. So the condition in which the soluble protein can efficiently express followed by preparation of the specific polyclonal antibody were explored. Methods Firstly, the cDNA of BcbZIP134 from Bupleurum chinense was expressed in Transetta (DE3) E. coli. Different concentrations of IPTG (0.05 and 0.5 mmol/L) and different culture temperatures (16 and 37 °C) were explored for efficient expression of target protein. Then, the expressed protein with His Tag was purified using Ni Sepharose 6 Fast Flow. Different concentrations of imidazole elution (15, 60, and 300 mmol/L) were used to elute the target protein. The purified protein of BcbZIP134 was used to immunize rabbit. Using the purified polyclonal antibody, the expression of BcbZIP134 in transgenic B. chinense hairy root lines was analyzed by Western Blot. Results Under the following conditions: IPTG 0.5 mmol/L, 16 °C, and overnight, recombinant protein of BcbZIP134 was obtained in high content using pEASY?-Blunt E1 vector and Transetta (DE3) E. coli. Anti-serum against BcbZIP134 was obtained with a high titer of 1: 51 200 after immunization in rabbit. The polyclonal antibody could react with BcbZIP134 overexpressed in transgenic B. chinense hairy root lines. Conclusion An efficient protein expression system in E. coli was constructed and the soluble recombinant protein for BcbZIP134 was obtained. By using this protein, the specific and high-performance polyclonal antibody was prepared. Furthermore, by using BcbZIP134 overexpressed hairy roots, the availability of the polyclonal antibody was verified by Western Blot analysis.

Moisture sorption and diffusion determination of Chinese Herbal Granules: moisture-resistant effects of fluidized bed granulation with dextrin

2018/8/3 0:00:00

Objective To investigate the effects of fluidized bed granulation with dextrin on moisture sorption and diffusion of Zexie Decoction granules. Methods The particle characterization was studied by the particle size, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and Fourier transform infrared (FTIR). The moisture sorption isotherm, equilibrium moisture content (EMC), and moisture diffusion coefficients were determined by using the saturated salt solution method. Results The particle size increased from 6.04 μm (powder) to 1201.47 μm (granule). The glass transition temperature of dextrin, Zexie Decoction powder, and granule was 107.13 oC, 94.82 oC, and 126.25 oC. As the increase of temperature, the initial rate of moisture sorption become higher. Furthermore, the initial rate of moisture sorption of Zexie Decoction granules was lower than those of powders and dextrin. The EMC and moisture diffusion coefficients were reduced significantly after granulation (p<0.01). Critical relative humidity and diffusion activation energy of granules were higer than powders. Conclusion Results suggested that fluidized bed granulation with dextrin could reduce the hygroscopicity of the Zexie Decoction extract powders and inhibite moisturediffusion, which is mainly related to the microstructure reorganization by fluidized bed granulation and anti-plasticizing effects of dextrin.

Zhengtian Pills accelerated long term potentiation both in schaffer collateral -CA1 and perforant path-dentate gyrus synapses

2018/8/3 0:00:00

Objective To investigate the effects of Zhengtian Pills (ZTP) on long term potentiation (LTP) both in schaffer-CA1 in vitro and perforant path-dentate gyrus (PP-DG) synapses in vivo. Methods Sprague-Dawley rats were randomly divided into five groups: control, positive control, migraine model, low-, and high-dose ZTP groups. Glyceryl trinitrate (10 mg/kg) was injected subcutaneously to make migraine rat model. Flunarizine (0.9 mg/kg) was set as positive control. Extracellular recording technique in vivo was used to record the effects of ZTP on LTP of PP-DG pathway in anesthetized rats; Using extracellular recording technique in vitro, the effects of ZTP on LTP of Schaffer Collateral-CA1 pathway in rat hippocampal slices were investigated. Results Compared to the control group, ZTP (1.08 g/kg) had significantly enhanced population spike amplitude in PP-DG pathway; Glyceryl trinitrate (10 mg/kg) significantly reduced population spike amplitude in PP-DG pathway; Neither ZTP (0.54 g/kg) nor Flunarizine (0.9 mg/kg) had significant effects on LTP in PP-DG pathway. Compared to the model group, ZTP (1.08 g/kg), ZTP (0.54 g/kg), and flunarizine (0.9 mg/kg) had significantly enhanced population spike amplitude in PP-DG pathway. Compared to the control group, ZTP (1.08 g/kg) significantly enhanced field excitatory postsynaptic potential (fEPSP) slope in Schaffer Collateral-CA1 pathway; Glyceryl trinitrate (10 mg/kg) significantly reduced fEPSP slope in Schaffer Collateral-CA1 pathway; Neither ZTP (0.54 g/kg) nor flunarizine (0.9 mg/kg) had significant effects on LTP in Schaffer Collateral-CA1 pathway, Compared to the model group, ZTP (1.08 g/kg), ZTP (0.54 g/kg), and Flunarizine (0.9 mg/kg) had significantly enhanced fEPSP slope in Schaffer Collateral-CA1 pathway. Conclusion: Combined with the previous study, the results gave a clue that the effects of ZTP on TRPV1 and hippocampal LTP or their interactions could be the important molecular mechanisms of ZTP acting as migraine and headache medication.

Pharmacokinetic study of ginsenoside Re after vaginal administration in rabbits by UPLC-MS/MS determination

2018/8/3 0:00:00

Objective A selective and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was employed to study the pharmacokinetics of ginsenoside Re (GRe) in rabbits after vaginal administration of Xiaomi suppository to evaluate the systemic exposure of the suppository for the local treatment. Methods Chromatographic separation was on an ACQUITY UPLC? BEH C18 column, and acetonitrile-0.1% formic acid was used as mobile phase in gradient elution. The plasma samples were deproteinized by acetonitrile, and the pharmacokinetic parameters were calculated by Winnonlin 6.4. Results Calibration curve of GRe showed a good linearity over the concentration range from 5 ng/mL to 500 ng/mL (r = 0.9999). The low limit of quantification of 5 ng/mL could satisfy the experimental requirement. The intra-day and inter-day precision of GRe at three concentrations were less than 1.96%, and the average recoveries of GRe were >64.0%. The pharmacokinetic parameters for vaginal administration were as follows: Tmax, 0.5 h; Cmax, 20.88 ng/mL; AUC0-t, 64.71 h·ng/mL and the residence time was 3.06 h. By using deconvolution calculation method, the cumulative absorption fraction of GRe was about 0.89%. Conclusion The systemic exposure of GRe was precious little after the vaginal administration of Xiaomi suppository.

Preparation and evaluation of Paclitaxel and Brucea Javanica oil core-matched nanoemulsions to treat cancer in vitro and in vivo

Shu-qing Cao1, Kuan-yun Zhang1,2, Xia Yan1 and Yan Ma1 — 2018/8/3 0:00:00

Objective Developed the core-matched nanoemulsions (CMNEs) to co-delivery paclitaxel-oleic acid prodrug (PTX-OA) and brucea javanica oil (BJO) for increasing the antitumor effect.. Methods Antitumor effects and mechanism of PTX-OA/BJO CMNEs that the combination therapy which based on core-matched technology (CMT) were evaluated in vitro and in vivo. Results The PTX-OA/BJO CMNEs were of nanoscale particle size (108.7 ± 2.3) nm and with entrapment efficiency of > 95%. The PTX-OA/BJO CMNEs displayed concentration and time-dependent cytotoxicity against HepG-2 cells and increased G2/M phase block. More importantly, a significant reduction of the tumor volume with no obvious toxicity was observed in nude mice model following administration of PTX-OA/BJO CMNEs compared with the control treated with normal saline (P < 0.05), which suggested the excellent efficacy in vivo. It was further found that the enhanced effectiveness of PTX-OA/BJO CMNEs were associated with the ability of inducing apoptosis of the tumor cells, as well as obviously inhibiting tumor cell proliferation and the activity of TOPO Ⅱ. Conclusion Co-encapsulation of two drugs with different mechanisms allows simultaneous interruption of diverse anticancer pathways, resulting in increased therapeutic response and lower toxicity.

Extraction and verification of miRNA from ginseng decoction

2018/8/3 0:00:00

Objective To verify the existence of microRNAs (miRNAs) extracted from fresh ginseng decoction. Methods Fresh ginseng was prepared into decoction according to the conventional method. The miRNA were extracted from the condensed ginseng decoction by plant microRNA extraction kit. Then miRNA were treated by DNase I and subjected to agarose gel electrophoresis and Agilent 2100 bioanalysis. MiR-159 and miR-6135, which were highly expressed in ginseng, were selected and verified by real-time quantitative PCR to detect the expression in the decoction. Results Ginseng miRNA were successfully extracted from fresh decoction. MiR-159 and miR-6135 were expressed in fresh decoction with lower levels than those of fresh ginseng. Conclusion miRNAs stably existed after processing, and retained some stability after high-temperature treatment. The findings provide a valuables basis for the further studies on ginseng miRNAs.

Synthesis and anti-tumor activities of novel 7-O-amino acids chrysin derivatives

2018/8/3 0:00:00

Objective To design and synthesize a series of chrysin derivatives and evaluate the antitumor activities with MTT assay, so as to investigate molecular structure-activity relationship with molecular docking. Methods Target products were synthesized with high yield by substitution reaction, hydrolysis reaction, esterification reaction, and saponification reaction in sequence, and activities of all compounds were evaluated with human gastric carcinoma cell lines MGC-803 and human breast carcinoma cell lines MCF-7 through standard MTT assay. Molecular docking results were calculated with Surflex Geom X programme of Sybyl X-2.0 version workstation. Results 7-O-amino acids chrysin derivatives 6a-6l were synthesized and their inhibitory effects were evaluated by comparing the material chrysin with positive control drug 5-fluorouracil (5-FU). Among these derivatives, compound 5b (IC50 = 24.50 ± 2.26 μmol/L), 5k (IC50 = 24.30 ± 2.19 μmol/L), and 6f (IC50 = 24.61 ± 2.01 μmol/L) showed better inhibitory activities against MGC-803 cell lines, and compound 5g (IC50 = 13.15 ± 1.73 μmol/L) and 5j (IC50 = 12.34 ± 1.25 μmol/L) showed better inhibitory activities against MCF-7 cell lines than chrysin and 5-FU. Molecular docking scores showed a credible consistency compared with MTT results. Conclusion Compounds 5b, 5d, 5g, 5j, 5k, and 6f showed good antiproliferative effects on specific tumor cells, and compound 5g should be researched further when according to molecular docking.

Synergistic Effect and Different Toxicities of Adjuvant Components of Realgar-Indigo naturalis Formula

2018/5/16 0:00:00

Objective Realgar-Indigo naturalis formula (RIF) is a well-known arsenic-containingpreparationthat is used to treat acute promyelocytic leukemia (APL) in China. In recent multicenter clinical trials, complete remission rates in APL patients have ranged from 96.08% to 100%. RIF has a satisfactory therapeutic effect, but its safety isa widespread concern, since the preparation contains arsenic, a wide-rangingand naturally occurring toxicant. In this study, in order to determine the toxic potential of RIF, acute toxicity and subchronic toxicity assayswere performed to evaluate the toxic potential of realgar and the adjuvant components of RIFin addition to realgar’s synergy with theseadjuvant components. Methods KM mice and Wistar rats were selected forthese experiments. To evaluate acute toxicity, the toxic effects of a single dose ofa gradient of concentrations of realgarwere firstly determined. Then, the toxic effects of combinations of gradient doses of realgar and fixed doses of I. naturalis and S. miltiorrhizawere evaluated.Results The results showed that when realgar was used alone, the LD50 was 2756.73 mg/kg (equivalent to 23.6mg/kg As2O3).However, theLD50dropped to 936.90 mg/kg when realgar was used with I. naturalis.By contrast, the LD50increased to 7538.86 mg/kg when realgar was used with S. miltiorrhiza. Hence, I. naturalis strengthened the toxicity of realgar, whereas S. miltiorrhizadisplayed the opposite effect. The subchronic toxicity assessment results revealed a trend that was consistent with acute toxicity.Changes in the levels of different valence states of arsenic were also taken into account.The test results of the effects of in vitro combinations of realgar and adjuvant components on soluble arsenic dissolution showed that I. naturalis increased the level of soluble arsenic in realgar extractsand I. naturalis suspensions when the realgar/I. naturalis ratio was 2, 1.5, and 1.0.However, S. miltiorrhizadid not affect it. Conclusion Based on the collective experimental results presented here, it can be concluded that the toxicity of RIF is the result of the soluble arsenic in realgar and that the I. naturalis and S. miltiorrhiza in the RIF exert completely opposite effects on the toxicity of realgar. This maybe an intelligent explanation for the compatibility of this formula, and this RIF study may therefore be viewed as a classic case of traditional Chinese medicine research on compatibility.

Toxicity-efficacy converting of ginseng combined with Fuzi Banxia incompatibility in heart failure stage of Cor pulmonale

2018/5/16 0:00:00

Objective Fuzi Banxia is one of eighteen antagonisms, previous studies have shown that the incompatibility could play special effects in the specific condition of diseases and appropriate compatible environment. The present study aims to evaluate the toxicity-efficacy of ginseng combined with Fuzi Banxia incompatibility intervening in the heart failure stage of cor pulmonale and to explore its mechanism. Methods Monocrotaline (MCT)-induced cor pulmonale were used in this study. Ultra high-resolution small animal ultrasound real-time imaging system and the right heart catheterization were used to estimate cardiac function. Semi automatic biochemical analyzer was used to test myocardial enzyme LDH, CK, and CK-MB in serum. The heart tissues were stained with HE, and TUNEL assay was used to assess the pathomorphological changes and myocardial apoptosis. The expression of hypertrophy and apoptosis associated genes: ANP, BNP, β-MHC, Bax, and Bcl-2 in the right ventricle were determined by RT-PCR. Results Fuzi Banxia combined with ginseng obviously attenuated mortality, decreased RVHI, and increased cardiac index; RVSP and mPAP were significantly reduced, and EF and FS were raised obviously; Myocardial enzymes LDH, CK, and CK-MB were pronounced attenuated; heart diameter reduced, right ventricular dilatation was significantly decreased, inflammatory cell infiltration notably reduced, and cardiac apoptosis rate was decreased obviously. Meanwhile the expression of hypertrophy-related ANP, BNP, and β-MHC mRNA were up-regulated, the expression of apoptosis-related Bax mRNA was down-regulated, and the expression of anti-apoptosis-related Bcl-2 mRNA and Bcl-2/Bax ratio were up-regulated. Conclusion Ginseng compatible environment could attenuate cardiac toxicity of Fuzi Banxia incompatibility intervening in the heart failure stage of cor pulmonale, and improve cardiac function, which may be related to the expression of hypertrophy and apoptosis associated genes, and thus delay the occurrence and development of heart failure.

Larvicidal Potential of Selected Indigenous Lichens against Three Mosquito Species- Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi

2018/5/16 0:00:00

Objective Mosquitoes are the major transmitting vectors of serious human diseases, causing millions of deaths every year with undesirable effects, including toxicity to non-target organisms. Some plants with insecticidal properties have been used in recent years for the control of a variety of pest insects and vectors. In the quest for alternative natural biological control agents against mosquito larvae lichens were selected. Method Larvicidal activity was assessed with methanolic extracts of Parmotrema reticulatum, Parmotrema kamatti, Parmotrema tinctorum, Parmelia erumpens, Leptogium papilosum,and Roccella montagnei against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. The standard WHO protocols with minor modifications were adopted and the bioassay was evaluated at the concentrations of 100-500 μg/mL for each lichen. Since all the lichen extracts showed complete mortality against C. quinquefasciatus in 100 μg/mL, the concentrations were decreased to 100, 50, 25, 12.5, and 6.25 μg/mL for C. quinquefasciatus. Larval mortality was observed for 24 h after treatment. Results All the lichen extracts exhibited activity against third instar larvae of A. aegypti and A. stephensi at 100 μg/mL, and 100% mortality was observed against the vector C. quinquefasciatus at 100 μg/mL. The highest larvicidal activity was found with L. papilosum against A. aegypti (LC50 = 81.127 μg/mL) and A. stephensi (LC50=89.10 μg/mL). Similarly, P. tinctorum and R. montagnei when tested against C. quinquefasciatus with minimum concentration <100 μg/mL exhibited significant activity with LC50 valuesof 5.32 and 6.97 μg/mL. Conclusion The bioassay results revealed larvicidal potential of lichens especially against C. quinquefasciatus with high mortality even at lower concertation. Hence, lichens can be used as an ideal sustainable approach for the control of lymphatic filariasis caused by vector C. quinquefasciatus.

Inhibition of metabolic disorders in vivo and in vitro by main constituent of Coreopsis tinctoria

2018/5/16 0:00:00

Objective To investigate the effects of the ethyl acetate extract of Coreopsis tinctoria (EAEC) on insulin resistance (IR) in rats fed a high-fat diet. Methods Male Sprague-Dawley (SD) rats were fed a HFD (60% fat) supplemented with EAEC for 8 weeks. The administration of EAEC to the rats with HFD-induced insulin resistance reduced hyperglycemia, plasma levels of insulin, and steatosis in the liver. Metabolomic study was used to analyse the metabolic levels of the high glucose-treated cells, control cells and marein-treated cells. Results High glucose and high fat conditions caused a significant increase in blood glucose, insulin, serum TC, TG and LDL-C levels, leading to abnormal IR in rats. However, treatment with EAEC protects against HFD-induced IR by improving the fasting serum glucose homeostasis and lipid homeostasis. The high glucose conditions significantly decreased glycogen synthesis and increased PEPCK, G6Pase and Krebs cycle-related enzyme protein levels, leading to an abnormal metabolic state in HepG2 cells. However, treatment with marein improved IR by increasing glucose uptake and glycogen synthesis and by downregulating PEPCK and G6Pase protein levels. The statistical analysis of the HPLC/MS data demonstrated that marein could restore the normal metabolic state. Conclusion The results revealed that EAEC ameliorates IR in rats, and marein has the potential to improve IR by ameliorating glucose metabolism disorders.

Salidroside shows a particular pharmacokinetic property in the model rats of myocardial ischemia

2018/5/16 0:00:00

Objective Salidroside showed potential pharmacological effect on plateau hypoxia and cardiovascular disease like myocardial ischemia. However, pharmacokinetic differences have not been assessed between the pathological model and the normal animals. This study focused on evaluating the pharmacokinetic properties of salidroside in animals with myocardial ischemia. Methods A reproducible and sensitive method was established and optimized based on liquid chromatography tandem mass spectrometry (LC–MS/MS) to determine salidroside in rats plasma. The data showed the AUC0–∞ and Cmax of salidroside proportionally increased along with dose elevation after singly intragastric administration of salidroside at a dose of 20, 50, and 100 mg/kg. Results Compared to the single dose, the Cmax, and AUC0–8h of salidroside markedly decreased while CL/F and V/F increased after multiple dosing. However, the Cmax and AUC0–8h of ischemic model rats were 0.35 and 0.39 fold lower than those in normal rats after a single dose at 50 mg/kg, with an increased CL/F and V/F. Surprisingly, after a consecutive administration of salidroside for 7 d, the mean Cmax, AUC0–8h increased 2.89 and 2.61 fold higher than a single dose in model rats, and even 2.28 and 4.03 fold higher than the normal controls after multiple doses. All the above fold values were statistically different (P < 0.01). Conclusion The particular PK properties of salidroside in ischemic model rats were presented in our study for the first time, suggesting that myocardial ischemia greatly affected pharmacokinetics exposure of the orally administrated salidroside after a single or multiple doses.

Identification and quality analysis of Panax notoginseng and Panax vietnamensis var. fuscidicus through integrated DNA barcoding and HPLC

2018/5/16 0:00:00

Objective Root or rhizome of Panax notoginseng (Sanqi) is known for its eutherapeutic effects. Panax vietnamensis var. fuscidicus, called Yesanqi or Yuenan sanqi by local residents, is also commercially available. They are similar in morphology, leading to serious safety problems in clinical medication. It is necessary to find the rapid and efficient methods to identify them. Methods P. notoginseng and P. vietnamensis var. fuscidicus were identified by DNA barcoding based on the ITS2 sequence. Notoginsenoside R1 and ginsenosides (Rb1, Rg1, Re, Rd, Rc, and Rb2) were analyzed in the roots, fibrils, stems, leaves, and flowers of P. notoginseng and P. vietnamensis var. fuscidicus using high-performance liquid chromatography (HPLC). Results P. notoginseng and P. vietnamensis var. fuscidicus were separated into branches of divergent clusters, and P. vietnamensis var. fuscidicus and Panax vietnamensis were clustered into a clade with 98% similarity according to DNA barcoding analysis. The chemical compositions of P. notoginseng and P. vietnamensis var. fuscidicus were similar in roots; while their compositions and contents of the notoginsenoside R1 and ginsenosides in flowers, leaves, stems, and fibrils were different. Conclusion ITS2 is a rapid and efficient method to identify P. notoginseng and P. vietnamensis var. fuscidicus. HPLC analysis indicated that pharmacological action might be different between P. notoginseng and P. vietnamensis var. fuscidicus.

Molecular Mechanism of Quercitrin on Osteogenic Differentiation and Adipogenic Differentiation of Rat Bone Marrow Stromal Stem Cells（rBMSCs）

2018/5/16 0:00:00

Objective The study was designed to investigate the molecular mechanism of quercitrin on osteogenic differentiation and adipogenic differentiation of rBMSCs. Methods rBMSCs were harvested from SD rats, and determination of alkaline phosphatase (ALP) activity, quantification of mineralization by Alizarin Red S staining, and the mRNA expression of osteogenic differentiation markers (Runx2, BMP-2, and OSX) by RT-PCR after rBMSCs stimulated by osteogenic induction with (0.1-10) μg/mL of quercitrin, quantification of Lipid droplet by Oil Red O staining and the mRNA expression of adipogenic differentiation marker (PPARγ, C/EBPα, and aP2) by RT-PCR after rBMSCs stimulated by adipogenic induction with (0.1-10) μg/mL of quercitrin. Results Quercitrin can up-regulate the mRNA expression of osteogenic differentiation markers (Runx2, BMP-2, and OSX) and increase ALP activity and mineralization after osteogenic induction, on the other hand quercitrin can suppress the mRNA expression of adipogenic differentiation markers (PPARγ, C/EBPα, and aP2) and decrease lipid droplet after adipogenic induction. Conclusion This study suggested that quercitrin not only stimulated osteogenic differentiation but also inhibited adipogenic differentiation of rBMSCs, which was associated with the up-regulation of Runx2, BMP-2, and OSX mRNA expression and the down-regulation of PPARγ, C/EBPα, and aP2 mRNA expression.

Protection of Shuxuetong combined with aspirin against cerebral ischemia/reperfusion injury through ameliorating coagulation and fibrinolysis system

2018/5/16 0:00:00

Objective To explore the influence of the combination of Shuxuetong (SXT) and aspirin on coagulation and fibrinolytic system of rats. Methods Suture method was applied to establish focal cerebral ischemia-reperfusion injury models in rats. A total of 50 SD rats were randomly divided into sham group, middle cerebral artery occlusion/reperfusion (MCAO/R) group, aspirin (10 mg/kg) group, SXT (0.6 mL/kg) group, and SXT (0.6 mg/kg) + aspirin (10 mg/kg) group. The neurological deficits were assessed according to Longa's grade 5 scoring method. The cerebral edema was detected by measuring the content of water in brain tissue. The volume of cerebral infarction was observed by 2,3,5-Triphenyltetrazolium chloride (TTC) staining. Blood plasma was collected by abdominal aortic method to test maximum platelet aggregation rate and four blood coagulation. CD61, CD62p, 6-keto prostaglandin F1α (6-keto-PGF1α), antithrombin III (AT-III), D-dimer, plasminogen activator inhibitor-1 (PAI-1), tissue factor (TF), tissue plasminogen activator (t-PA), platelet thromboxane B2 (TXB2), and von Willebrand factor (vWF) content in rat plasma were detected by ELISA. Results SXT combined with aspirin could improve the neurological deficits, alleviate cerebral edema, and decrease the cerebral infarct value. Compared with the sham operation group, fibrinogen (FIB), 6-PGF1α, AT-III, and t-PA in model group were significantly decreased; Compared with the model group, the above-mentioned indexes in SXT and aspirin treatment group were significantly increased. The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), D-dimer, PAI-1, TF, TXB2, and vWF of the model group were significantly increased; the above-mentioned indexes in blood SXT + aspirin treated group were significantly decreased. There was a significant difference between the combined group and SXT group. The maximum concentration of plateletsin aspirin treated rats was significantly decreased, however, MPAR was reversed in SXT + aspirin treated group. Conclusion SXT combined with aspirin can effectively inhibit platelet activation, regulate the maximum concentration of platelets, and improve coagulation function and fibrinolysis system.

Effect of high exposure of chlorogenic acid on lipid accumulation and oxidative stress in oleic acid-treated HepG2 cells

Ya-yun Liu, Ting Zhai, Qing-qing Yu, Jing Zhu and Yong Chen — 2018/5/16 0:00:00

Objective To evaluate the effect of high concentration of chlorogenic acid (CGA) on non-alcoholic fatty liver disease (NAFLD) in normal and oleic acid (OA) treated HepG2 cells, as well as the underlying mechanism involved in the fat accumulation, oxidative stress and insulin resistance (IR) induced by CGA treatment. Methods OA (0.5 mmol/L) induced hepatic steatosis was established in HepG2 cells as an in vitro model of NAFLD. The normal and OA-treated HepG2 cells were treated by CGA (0, 0.5, 1, and 2 mmol/L) for 24 h, then cellular lipid droplets, reactive oxygen species (ROS), and glucose uptake were evaluated by Oil Red O staining and cellular biochemical assays, respectively. Signaling pathways involved in?adipogenesis including SREBP-1c and PNPLA3, oxidative stress, and IR including CYP2E1 and CYP4A, were investigated by Western blot and RT-qPCR. Results CGA (0.5, 1, and 2 mmol/L) treatment increased the cellular lipid droplets and the expression of SREBP-1c and PNPLA3 in the tested cells. Additionally, 2-NBDG uptake was significantly increased, whereas the cellular ROS and protein levels of CYP2E1 and CYP4A were significantly decreased in OA-treated cells. Conclusion Our results suggest that high concentrations of CGA ameliorate OA-induced oxidative damage and IR likely by inhibiting the expression of CYP2E1 and CYP4A, promote lipid accumulation by inducing the expression of SREBP-1c and PNPLA3 in the tested cells.

Comparative Brain Pharmacokinetic Study of Jiaotai Pills in Normal and Insomnic Rats using Brain Microdialysis Combinated with LC-MS/MS

2018/5/16 0:00:00

Objective To compare the brain pharmacokinetics of five protoberberine-type alkaloids (i.e. berberine, palmatine, coptisine, epiberberine, and jatrorrhizine), which were the main bioactive constituents of Jiaotai Pills (JTP), in normal and insomnic rats orally administrated with the JTP. Methods The detection was conducted by a fully validated liquid chromatography-tandem mass spectrometry combinated with brain microdialysis method. Brain microdialysis probes were inserted into the hippocampus of rats. JTP extracts were administrated intragastrically and then brain microdialysates were collected at 30 min time intervals for 10 h. The separation of the five protoberberine-type alkaloids was carried out on a BDS Hypersil C18 column using a mobile phase consisting of acetonitrile and water (containing 5 mmol ammonium acetate adjusted to pH 5.0) within 4 min. The quanti?cation was performed by multiple reaction monitoring with the transitions of m/z 336.0-320.1 for berberine, m/z 352.0-336.1 for palmatine, m/z 338.0-322.1 for jatrorrhizine, m/z 336.0-320.1 for epiberberine, m/z 320.0-292.1 for coptisine and m/z 356.4-192.1 for IS. Results The lower limit of quanti?cation for five protoberberine-type alkaloids was 0.05 ng/mL. Linearity, accuracy, precision, stability and matrix effect of five analytes were all satisfactory. Five protoberberine-type alkaloids were quickly distributed in the brain. Moreover, signi?cant differences in the principal pharmacokinetic parameters such as AUC and T1/2 of the analytes were observed between two groups. Conclusion The LC-MS/MS method combinated with microdialysis is useful in the brain pharmacokinetic study of five protoberberine-type alkaloids. The results indicated that the rates of analytes absorption in insomnic rats were signi?cantly higher than those in normal rats. Besides, the protoberberine-type alkaloids could bring a direct effect on the neuron in the hippocampus.

Hederacolchiside A1 suppresses proliferation of tumor cells by inducing apoptosis through modulating PI3K/Akt/mTOR signaling pathway

2018/5/16 0:00:00

Hederacolchiside A1, exhibits cytostatic and cytotoxic activity against various cancer cells in vitro, however, the mechanism is not well understood. In this study, Hederacolchiside A1 from Pulsatilla chinensis was isolated and tested its anticancer activity and mechanism. Hederacolchiside A1 could inhibit proliferation of A549, SMMC-7721, BEL-7402, and MCF-7 cells by MTT assay. Investigations of apoptosis of treated cancer cells were identified in hederacolchiside A1 by flow cytometric analysis of annexin V expression. Based on the results of western blotting and JC-1 staining, hederacolchiside A1 reduced the mitochondrial membrane potential and Bcl-2 protein levels, whereas cleaved caspase-3 was higher. Furthermore, hederacolchiside A1 effectively inhibited the phosphorylations of phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt), and mammalian target of rapamycin (mTOR). In vivo study showed that hederacolchiside A1 (3.0, 4.5, and 6.0 mg/kg, ip) could significantly inhibit the weight of tumor in an H22 xenograft model. Similar inhibitory activities were observed when the compound (3.25, 7.5, and 15.0 mg/kg, ig) was tested in nude mice xenograft tumor models using human breast carcinoma MCF-7 cells. These data indicated that hederacolchiside A1 suppressed the proliferation of human tumor cells by inducing apoptosis through modulating the PI3K/Akt/mTOR signaling pathway.

Protective effects of combination of Xuesaitong and aspirin on cerebral ischemia and reperfusion injury in rats

2018/5/16 0:00:00

Objective To investigate the protective effects of the combination of Xuesaitong (XST) and aspirin on cerebral ischemia and reperfusion injury (CIRI) in rats, and further explore the underlying mechanisms. Methods A total of 150 male Sprague-Dawley (SD) rats were randomly divided into five groups with 30 rats in each group: sham group, middle cerebral artery occlusion/reperfusion (MCAO/R) model group, XST group, aspirin group, and XST + aspirin group. Rats were pretreated with XST, aspirin, or XST + aspirin for 7 d. One hour after the last administration, a model of CIRI was induced by MCAO/R. Neurological deficits were assessed using Longa’s five-point scale. Cerebral edema was detected by the measurement of brain water content. The volume of cerebral infarction was determined by 2,3,5-triphenyltetrazolium chloride (TTC) staining. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), as well as levels of malonaldehyde (MDA) were detected by commercial kits. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of interleukin-1 (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1), and kynurenine in serum, cerebral cortex, and hippocampus of MCAO/R rats. The protein expression of nuclear factor erythroid 2-related factor (Nrf2), heme oxygenase-1 (HO-1), I-kappa B alpha (IκBα), and nuclear factor kappa B (NF-κB)/p65 in the cortex were analyzed by western blotting. Results Treatment of XST, aspirin, and XST + aspirin significantly alleviated the neurological deficits, cerebral edema, and cerebral infarct volume induced by MCAO/R. Treatment of XST, aspirin, and XST + aspirin also reduced MDA, IL-1β, IL-6,TNF-α, MCP-1, and kynurenine levels, and increased SOD, CAT, GSH-Px, IL-4, and IL-10 levels in serum, cerebral cortex, and hippocampus of MCAO/R rats. Furthermore, treatment of XST, aspirin, and XST + aspirin decreased the expression of nuclear NF-κB/p65 and increased the expression of IκBα, nuclear Nrf2, and HO-1. Importantly, the combination of XST and aspirin enhanced the protective effects of XST or aspirin treatment alone on CIRI in rats. Conclusion The combination of XST and aspirin significantly inhibited oxidative stress and inflammation in serum, cerebral cortex, and hippocampus of MCAO/R rats. The combination of XST and aspirin exerted more protective effects than XST or aspirin treatment alone. The combination of XST and aspirin might provide the synergistic therapeutic effects on CIRI, and deserve further clinical investigation.

Ergosta-7,22-diene-2β,3α,9α-triol(EGDT) from Ganoderma lucidum inhibits nasopharyngeal carcinoma cells by blocking EGFR signaling pathway

Wen-wei Wu, Zi-jing Ye, Yi-mou Xu, Jin Zhang and Jin-senTang — 2018/2/5 0:00:00

Objective To investigate the efficacy and mechanism of EGDT against NPC cell lines. Methods MTT assay was used to assess cell proliferation inhibition of EGDT; The apoptotic induction and cell cycle arrest were detected by flow cytometry; Western blot was adopted to detect the protein levels; Quantitative Real-time PCR was used to determine the mRNA expressions; The NPC xenografts were established to evaluate the tumor growth inhibition of EGDT; Immunohistochemistry was applied to analyze the EGFR expression in the tumor tissues. Results EGDT showed proliferation inhibition on the NPC cell, induced G0/G1 phase arrest and cell apoptosis in vitro. EGDT decreased the protein and mRNA levels of EGFR and its downstream RAF/MEK/ERK and PI3K/AKTpathways in time- and dose-dependent manner. Furthermore, EGDT also showed a sound antitumor activity in NPC xenograft in vivo. Conclusion The treatment of EGDT displays EGFR and its mediated downstream signaling pathway blockade through decreasing the protein and mRNA levels, suggesting a promising strategy in treating human NPC.

Effect of Light Quality on Total Gypenosides Accumulation and Related Key Enzyme Gene Expression in Gynostemma pentaphyllum

2018/2/5 0:00:00

Objective Light quality has effect on the accumulation of gypenosides in the medicinal plant Gynostemma pentaphyllum in the family Cucurbitaceae, while the squalene synthase (SS) and squalene epoxidase (SE) are the key enzymes for gypenoside biosynthesis. The objective of this study was to elucidate the relationship between light quality and biosynthesis key enzyme involving the regulation of gypenoside accumulation. Methods The content of total gypenosides was measured by colorimetric method and the expression of SS and SE gene was determined by quantitative Real-time PCR in the seedlings of G. pentaphyllum which were grown with different light quality. Results Light quality showed remarkable impacts on the accumulation of total gypenosides. The highest content of total gypenosides was determined in the plant under red light condition, followed by blue light and white light, while the lowest content was recorded under dark condition. qRT-PCR analysis proved that the expression levels of SS and SE genes were also affected by light quality. The high-level gene expressions of SS and SE were found in the plant under red light condition, followed by blue light, with the least content in darkness. The statistical analysis revealed that the total gypenosides were significantly different in different light treatment and the content of total gypenosides was positively related to the expression of SS and SE genes. Conclusions Light quality regulates gypenoside accumulation via altering the expression of SS and SE in G. pentaphyllum.

Protective effect of icaritin on focal cerebral ischemic-reperfusion mice

2018/2/5 0:00:00

Objective The purpose of this study was to investigate the protective effects of icaritin (ICT), one of the active ingredients in Epimedii Folium, on mouse model of cerebral ischemia-reperfusion (I/R) in vivo. Methods ICR mice were subjected to an 1 h transient middle cerebral artery occlusion (MCAO) and followed by 24 h of reperfusion. Neurological deficits, infarct volume, brain edema and survive rate were measured, respectively. The levels of brain IL-1β, TNF-α, ROS and DNA-binding activity of NF-κB p65 were measured by ELISA kits. The levels of malondialdehyde (MDA) and activities of superoxide dismutase (SOD) were detected by spectrophotometry, and the release of nitric oxide (NO) were detected by Griess kit. Results The results showed that ICT markedly reduced the neurological deficit scores, brain edema, infarct volume and increased the survival rate of the cerebral I/R mice. The expression of IL-1β, TNF-α, NO, MDA and DNA-binding activity of NF-κB p65 were significantly inhibited by ICT, while the activity of SOD were up-regulated at the same time. Conclusion These results suggested that ICT possessed significant neuroprotective effects in cerebral I/R mice, which might be related to prevent neuroinflammatory and oxidative damage.

Protection of Salvianolate Lyophilized Injection combined with Xueshuantong Injection (Lyophilized) against focal cerebral ischemia/reperfusion injury in rats through suppression of inflammatory response

2018/2/5 0:00:00

Objective Inflammatory reactions induced by microglia in the brain play an important part in the pathogenesis of focal cerebral ischemia/reperfusion (I/R) injury, resulting in neuronal death. Salvianolate Lyophilized Injection (SLI) and Xueshuantong Injection (Lyophilized) (XST), which have been widely used in the treatment of acutely cerebral infarction clinically in China, exhibit various biological activities. In this study, we investigated the neuroprotective properties of SLI combined with XST in a rat model of middle cerebral artery occlusion- reperfusion (MCAO/R). Methods In this study, male Wistar rats were subjected to 1.5 h of middle cerebral artery occlusion followed by reperfusion for 24 h. The rats were randomly divided into the following six groups: normal group (NOR), model group (MOD), SLI group (21mg/kg, SLI), XST group (100mg/kg, XST), SLI combined with XST (XST 100 mg/kg: SLI 21 mg/kg, 1X1S), and Edaravone (as a positive control drug, 6 mL/kg, EDI), once a day for 3 d. We observed the neuronal injury, the expression of glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule 1 (IBA-1), and the changes of pro-inflammatory mediators interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) and anti-inflammatory mediator interleukin-10 (IL-10). Results 1X1S treatment significantly increased the number of neuron, compared with the MOD group, SLI group and XST group. Gliosis (GFAP and IBA-1) and expression of pro-inflammatory mediators IL-6 and TNF-α were significantly reduced. Meanwhile, 1X1S significantly increased the expression of anti-inflammatory mediator IL-10 in the brains of MCAO/R rats, compared with the MOD group, SLI and XST groups. SLI and XST also remarkably down-regulated the expression of IL-6 and TNF-α compared with the MOD group. Conclusions This study shows that SLI combined with XST (1X1S) can protect cerebral ischemia-reperfusion injury due to its anti-inflammatory property, and may provide a potential promising new therapeutic strategy for acute ischemic stroke.

Synthesis and Biological Evaluation of New Peroxo-bridged Diosgenin Derivatives

2018/2/5 0:00:00

Objective: In order to find lead compound with anti-HBV activity from peroxo-bridged diosgenin deriva- tives obtained with Eosin Y as the photosensitizer. Method: Eosin Y was used as the photosensitizer to activate the oxygen in the air to synthesize novel diosgenin derivatives with peroxo-bridge.The structures of synthesized compounds were identified by NMR and HR-MS. Their cytotoxicity and anti-hepatitis B activity were evaluated via MTS assay and ELISA method, respectively. Results: Six diosgenin derivatives were synthesized, three of which contain peroxo-bridge, and their structures were confirmed by spectroscopy. It showed that 5 α,8 α-peroxo-6-alkenyl-diosgenin( 7 ) could suppress the production of HBsAg on transfected HepG2.2.15 cells at low-toxic concentration and the inhibition rate (%) on HepG2.2.15 cells was 18.28% at 12.50 μg/mL, better than that of 3TC (7.30% at 12.50 μg/mL) and others. Conclusion: Due to its lower cytotoxicity and potential anti-hepatitis B activity, compound 7 could be developed as the promising candidate of anti-hepatitis B drug. It also indicated that the peroxo-bridged derivatives had potential biological values for developing clinical agents.

Analgesic and Anti-inflammatory Activities and Mechanisms of 70% Ethanol Extract of Zygophyllum macropodum in Animals

Xiao-rong Yanga,b, Xiang-feng Zhanga, Xue-mei Zhanga and Hong-yan Gaob,c — 2018/2/5 0:00:00

Objective: To screen the preliminary phytochemical components, to investigate the acute oral toxicity, the analgesic and anti-inflammatory effects, and to analyze inflammatory factors on mice or rats of 70% ethanol extract of Z. macropodum aerial parts (ZME). Methods: Preliminary phytochemical screening was carried out by colour reaction. Acute oral toxicity was investigated by body weight, relative organ weight and other toxic signs. Acetic acid induced writhing and hot plate test were used to determine analgesic effect. Acetic acid induced vascular permeability and carrageenan induced paw edema were used to confirm anti-inflammation. Protein in pleural effusion, prostaglandin E2 (PGE2) and tumor necrosis factor alpha (TNF-α) in serum of pleuritic rats induced by carrageenan were analyzed to explore the action mechanisms. The test groups received ZME with 100, 300, 600 mg/kg, the positive control with aspirin (ASP) 200 mg/kg for mice, and ZME with 70, 210, 420 mg/kg and ASP 150 mg/kg for rats orally. The control (C) or negative control (NC) groups received 2% Tween 80 of 10 mL/kg orally. Results: ZME contained flavonoids, saponins, phenols and tannins, steroids, terpenoids, fats and oils, glycosides, carbohydrates, and reducing sugar, but no alkaloids. The lethal dose 50% (LD50) of ZME was greater than 2000 mg/kg and no toxic or deleterious effects and death during 14 days. Oral administration 300 and 600 mg/kg of ZME produced analgesic and anti-inflammatory effects significantly (Pp < 0.05 or Pp < 0.001) vs. NC. It can reduce the writhing number, prolong the heat resisting time, suppress the permeability of the capillary wall increasing, mitigate the paw edema, reduce the content of protein in pleural effusion, and reduce PGE2 and TNF-α in blood. Conclusions: ZME possessed analgesic and anti-inflammatory activities which related to inhibit the production of protein, PGE2 and TNF-α. The LD50 of ZME treated orally to mice was greater than 2000 mg/kg.

Effects of water extracts of Davallia formosana on ovariectomized mice

Chia-Feng Wua,b, Cheng-Yu Chenb, Ming-Chang Wua and Jen-Shinn Lina — 2018/2/5 0:00:00

Objective: The Taiwanese native fern, Davallia formosana Hayata, is used to treat bone diseases in traditional Chinese medicine, but, very few animal tests to verify. To study how this fern and its active component alter bone metabolism. Methods: It was examined that the effects of low and high doses of D. formosana water extracts (DFW, 50 and 200 mg?kg-1?d-1) and [-] epicatechin-3-O-D-allopyranoside isolates of DFW (ECAP, 0.06 and 2.5 mg?kg-1?d-1) on protein levels of important indicators of osteoclast and osteoblast activities. Ovariectomized mice were used as a model of osteoporosis. Bone morphogenetic protein 2 (BMP-2), collagen 1 (COL-1), alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN) were analyzed using western blotting of femur tissue. Interleukin (IL)-1β and tumor necrosis factor (TNF)-α were measured in the serum, and hematoxylin and eosin staining was used to assess bone morphology. Results: Treatment with DFW or ECAP significantly induced BMP-2, COL-1, ALP, RUNX2, OCN, and OPN protein expression, indicating stimulation of osteogenesis. The treatments also reduced IL-1β and TNF-α expression, indicating inhibition of osteoclastogenesis. In histological examinations, mice treated with DFW or ECAP had more bone trabeculae. The results demonstrate that DFW and ECAP inhibit osteoclast differentiation and promote osteoblast differentiation, and effectively ameliorate ovariectomy (OVX)-induced osteoporosis. Conclusion: ECAP is a major component of DFW, which may have therapeutic potential for the treatment of diseases associated with excessive osteoclastic and insufficient osteogenic activity. ECAP may not be the specific active molecule because it is unstable in the stomach. However, the major active molecule needs to be confirmed using a relevant pharmacological model.

Metabolite Characterization of Penta-O-galloyl-??￠-D-glucose in Rat Biofluids by HPLC/QT of MS

2018/2/5 0:00:00

Objective To understand the in vivo metabolic fate of 1,2,3,4,6-Penta-O-galloyl-?-D-glucose (PGG) naturally existed in many medicinal herbal and food plants such as Rhus chinensis Mill, Paeonia suffruticosa, Paeonia lactiflora and Mango. Methods The metabolites of PGG in rat biofluids were characterized using high performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry (HPLC/QTof-MS). Results Ten metabolites in urine, five metabolites in feces and two metabolites in plasma, were observed when the rats were administrated with a single intravenous injection of PGG (20 mg/kg). Conclusion PGG is firstly metabolized to gallic acid, then gallic acid undergoes sulfation, glucuronidation and methylation by rat liver. The determination of metabolites and the proposed metabolic pathway of PGG in vivo will be benefit to gain deeper insights into its pharmacological activities.

Evaluation and Prediction of Ecological Suitability of Medicinal Plant American Ginseng (Panax quinquefolius)

Qin Zhanga, Jian Wenb, Zi-qian Changc, Cai-xiang Xiea and Jing-yuan Songa — 2018/2/5 0:00:00

Objective American ginseng is a medicinal plant with large market demand, however, its producing areas are shrinking because of the continuous cropping obstacles in China. Therefore, it is urgent to establish a suitable model to determine the new producing areas. Here we evaluated and predict the suitable areas of American ginseng using the maximum entropy model (MaxEnt). Methods Based on the 37 environmental variables over thirty years from 1970 to 2000 and 226 global distribution points of American ginseng, MaxEnt was used to determine the global ecological suitable areas for American ginseng. The Receiver Operating Curve (ROC) was used to evaluate the model prediction accuracy. Meanwhile, an innovative ecological variable, the precipitation–temperature ratio, was established to indicate the climate characteristic in the American ginseng suitable areas based on the monthly precipitation and temperature. Results The potential ecological suitable areas of American ginseng were primarily in Appalachian Mountain in America and Changbai Mountain in China, about in the range of 35°N-50°N, 60°W-120°W and 35°N-50°N,110°E-145°E, respectively, including the United States, Canada, China, North Korea, South Korea, Russia and Japan. South Korea and Japan were the potential producing regions. The precipitation–temperature ratios were stable at (0.22, 0.56) of the vigorous growth period (April to October) in the best suitable areas of American ginseng, serving as characteristic parameters to optimize the prediction model. The model showed that the common soil parameters were pH 4.5-7.2, Base Saturation (BS) above 80%, Cation Exchange Capacity (CEC) 10-20 cmol/kg, Organic Carbon (OC) ＜ 1.4%, and the soil types were sandy loam or loam. Conclusion An optimized MaxEnt model was established to predict the producing area for American ginseng that needs to be validated by a field test.

Comparative chemical characters of Sparganii Rhizoma from different regions in China

2018/2/5 0:00:00

Objective To compare the chemical characters of Sparganii Rhizoma from different areas via chromatographic analysis and to establish a sensitive LC/MS method for quality assessment of Sparganii Rhizoma. Methods Under the optimised HPLC-PDA chromatographic conditions, twenty batches of Sparganii Rhizoma were analyzed by HPLC fingerprints. Principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (OPLS-DA) and hierarchical cluster analysis (HCA) were performed based on all peak areas of Sparganii Rhizoma fingerprints. Meanwhile, part of common peaks were subsequently quantified by UFLC-QTRAP-MS. Results The similarity values of HPLC fingerprints fluctuated in a wide range of 0.511-0.973, which showed variable differences of chemical characters among Sparganii Rhizoma from twenty habitats. PCA, OPLS-DA and HCA indicated that samples could be divided into five groups with different chemical characters, which generally corresponded with their geographical distributions. A total of 31 peaks in HPLC fingerprints were marked, and eight of them were identified and quantified. The quantitative result was generally in agreement with the classifications based on HPLC fingerprints, which indicated that Sparganii Rhizoma samples from eastern China mostly contained more contents including phenolic acids and flavonoids. Conclusion This study not only proved that there are relationships between geographic distributions and internal chemical compositions of plants, which could provide evidence to the traditional Chinese medicine concept “geo-authentic”, but also supplied a sensitive and rapid simultaneous quantitive method for the quality estimation of Sparganii Rhizoma.

A phytochemical study on Dichocarpum auriculatum???an endangered medicinal plant peculiar to China

Pei Li, Shuang-shuang Liu, Qing Liu, Jei Shen, Chun-nian He and Pei-gen Xiao — 2019/11/12 9:40:23

Objective: Dichocarpum auriculatum, an endangered perennial herb, is endemic to China and has been used as folk medicines for the treatment of cough, hepatitis, scrofula, and epilepsy. However, there is no phytochemical report on this herbal so far. For the resource development and protective importance of this endangered medicinal plant, a phytochemical study was undertaken. Methods: The chemical constituents were purified by silica gel column, Sephadex LH-20 column, and semi-preparative reversed phase HPLC. NMR and MS spectra were used for structural identification. Results: Thirteen compounds were isolated from D. auriculatum. Their structures were characterized as jatrorrhizine (1), berberine (2), steponine (3), magnoflorine (4), coclauril (5), menisdaurin (6), menisdaurilide (7), aquilegiolide (8), (6R, 9S)-3-oxo-α-ionol-β-D-glucopyranoside (9), blumenol C glucoside (10), palmitic acid (11), dibutylphthalate (12), and auriculatum A (13). Conclusion: Compound 13 is a new diester terephthalate derivative. All the compounds are obtained from the genus Dichocarpum for the first time, and compounds 9 and 10 have potential chemotaxonomic significance to the genus Dichocarpum.

Gynura procumbens aqueous extract alleviates nonalcoholic steatohepatitis through CFLAR-JNK pathway in vivo and in vitro

2019/11/12 9:40:23

The pre-sent work was to in-ves-ti-gate the pro-tec-tive ef-fects of the aque-ous ex-tract of Gy-nura procum-bens (GPAE) against non-al-co-holic steato-hep-ati-tis (NASH) in mice and NCTC-1469 cells. Methods C57BL/?6J mice were fed with me-thio-n-ine and choline-de-fi-cient (MCD) diet and ad-min-is-tered si-mul-ta-ne-ously with GPAE (500 and 1000?mg/?kg/?d, re-spec-tively) by gav-age for six weeks. The bio-mark-ers of NASH in serum and liver were de-ter-mined. NCTC-1469 cells were pre-treated with 0.25?mmol/?L palmitic acid (PA) plus 0.5?mmol/?L oleic acid (OA) for 24?h or treated with ade-n-ovirus ex-press-ing short-hair-pin RNA against CFLAR (Ad-shCFLAR) for 24?h and then treated with GPAE (80 and 160?μg/?mL, re-spec-tively) for 24?h, and the con-tent of cel-lu-lar bio-mark-ers of NASH was de-tected. Results In mice treated with MCD, GPAE could de-crease the lev-els of serum ALT, AST, the con-tent of he-patic TG, TC and MDA, re-press the ac-tiv-i-ties and pro-tein ex-pres-sion of CYP2E1 and CYP4A and the phos-pho-ry-la-tion of JNK, in-crease the ac-tiv-i-ties of HO-1, CAT and GSH-Px, up-reg-u-late the mRNA ex-pres-sion of PPARα, FABP5, CPT1α, ACOX, SCD-1, mG-PAT, MTTP and the pro-tein ex-pres-sion of CFLAR and NRF2. In NCTC-1469 cells treated with PA and OA, GPAE could de-crease the con-tent of cel-lu-lar TG and ROS, pro-mote the up-take of 2-NBDG, up-reg-u-late the pro-tein ex-pres-sion of CFLAR and NRF2. In NCTC-1469 cells treated with Ad-shCFLAR, GPAE up-reg-u-lated the mRNA and pro-tein ex-pres-sion of CFLAR, down-reg-u-lated the phos-pho-ry-la-tion of JNK, and in-creased the pro-tein ex-pres-sion of NRF2 and pIRS1. Conclusion These re-sults in-di-cated that the ac-ti-va-tion on CFLAR-JNK path-way might be the main anti-NASH mech-a-nism of GPAE, which on the one hand pro-mote the β-ox-i-da-tion and ef-flux of fatty acids in liver, and fi-nally re-duce he-patic lipid ac-cu-mu-la-tion, on the other hand in-crease the ac-tiv-i-ties of anti-ox-i-dant en-zymes and in-hibit the ac-tiv-i-ties of ROS gen-er-a-tion en-zymes by ac-ti-vat-ing NRF2, and there-fore at-ten-u-ates he-patic ox-ida-tive stress dam-age.

Selection of quality markers of Jasminum amplexicaule based on its anti-diarrheal and anti-inflammatory activities: Effect-target affiliation-traceability-pharmacokinetics strategy

2019/11/12 9:40:23

Objective: To investigate therapeutic mechanism in Jasminum amplexicaule (Oleaceae) and verify its main active component as quality control markers Methods: Established mouse models of diarrhea, intestinal angina, and inflammation were first used to select herb fractions with optimum efficacy, followed by an in vitro experiment to determine key targets associated with effects of J. amplexicaule extract. Further, the selected fractions were isolated and purified, its components were identified, and the obtained compounds were verified for their effects on NF-κB and iNOS. Finally, effective compounds were administered to rats, their plasma pharmacokinetic parameters were calculated, and quality markers (QMs) reflecting therapeutic activities of J. amplexicaule were confirmed. Results: Trichloromethane and ethyl acetate fractions had significant anti-diarrheal, anti-inflammatory, and analgesic effects. The trichloromethane fraction also reduced BDNF, p38 MAPK, p-p38 MAPK, NF-κB p65, and p-NF-κB p65 levels in the ileum in a rhubarb-induced diarrhea mouse model. Additionally, it inhibited LPS-induced NF-κB transcription and nitric oxide (NO) production in RAW264.7 macrophages, which suppressed iNOS expression. Therefore, the trichloromethane fraction was further investigated. QMs candidate selection identified 17 compounds, and results of in-vitro therapeutic validation indicated that methyl caffeate and isochlorogenic acid B had the strongest anti-diarrheal, anti-inflammatory, and analgesic activities. After being validated by a UHPLC-MS-MS method, concentrations of these target compounds were accurately determined in the rat plasma and pharmacokinetic parameters were calculated. Cmax, Tmax, and T1/2 were respectively 575.35 ng/mL (2.963 nmol/mL), 0.5 h, and 0.45 h for methyl caffeate and 262.03 ng/mL (0.5034 nmol/mL), 0.25 h, and 2.03 h for isochlorogenic acid B. Because these candidate compounds exhibited favorable pharmacokinetics, they were considered as QMs of J. amplexicaule. Conclusions: The present study accurately and effectively identified QMs of J. amplexicaule that act as indicators of efficacy and quality.

Hypoglycemic and antioxidant activities of Clerodendrum inerme leaf extract on streptozotocin-induced diabetic models in mice

2019/11/12 9:40:23

Objective: Recently, much attention has been paid to natural product-derived compounds for antidiabetic drug discovery. More recent studies are being focused on clarifying the bioactivity of plants and derived products. The aim of the present study was to investigate the anti-oxidant and antidiabetic activities of Clerodendrum inerme leaf extract (CILE) in streptozotocin-induced diabetic mice. Methods: C. inerme leaves were analyzed for preliminary phytochemical properties and the content of total phenolic and flavonoid were determined. In vitro antioxidant activity was measured using DPPH assay. Streptozotocin-induced diabetic model in mice was applied for in vivo study by the effect of CILE at two dose levels (343 and 686?mg/kg b.w.). Results: The results showed that C. inerme leaves contained the major constituents of flavonoids, alkaloids, tannins, triterpenes, and saponins. CILE exhibited the total polyphenol and flavonoid content with 120.458 mg gallic acid equivalent/g dry weight and 4.494 mg hispidulin equivalent/g dry weight, respectively. The anti-oxidant activity of CILE was expressed with IC50 = 25.28 μg/mL. CILE at the doses of 343 mg/kg and 686 mg/kg after 7 d administration exerted a decrease in plasma glucose, protected the liver, kidneys against oxidation stress via increasing glutathione content in the liver, and reduced malondialdehyde content in the liver and kidneys. Pancreatic histological analysis in diabetic mice treated with CILE also showed the pancreatic β-cells regeneration via increasing the size and number of pancreatic islets. Conclusion: These findings suggested that C. inerme leaves have potent antidiabetic and anti-oxidant activities. The results provide reliable scientific base, which is the premise for further research and development of CILE as supplements.

In vitro anti-Helicobacter pylori activity of aqueous extract from Persian Oak testa

2019/11/12 9:40:23

Background: Growing problem of antibiotic resistance in Helicobacter pylori, as a common cause of chronic gastritis and even stomach cancer, demands searching for novel candidates of herbal sources. Objective: This study is aimed at assessing the antimicrobial activity of aqueous extract obtained from Quercus brantii var. persica seed coat (Testa) on H. pylori isolated from gastric biopsy specimens. Methods: Such specimens were collected from 100 patients presenting with endoscopic gastroduodenal findings. Testa extracts were prepared from Persian oak forests in the province of Kohgiluyeh and Boyer-Ahmad, IRAN. H. pylori isolates were obtained by a series of standard bacteriology tests and cell culture, then were confirmed by PCR. The activity of testa extracts towards 25 H. pylori isolates was assessed by well diffusion method, microdilution assay, and a disk diffusion assay in vitro. Results were analyzed statistically by one-way ANOVA analysis. Results: Aqueous extract of testa demonstrated an antimicrobial activity with zone diameters of inhibition ranged from 0 mm to 40 mm. Its inhibitory activity increased simultaneously with increasing extract concentration. The lowest MIC and MBC were both recorded as 2 μg/mL. Anti-H. pylori activity of testa extract was approximately close to tetracycline and metronidazole and less than amoxicillin. A potent extract of testa possessed significant inhibitory activity (P < 0.05). Conclusion: Testa extract is suggested as a natural therapeutic source against the gastric H. pylori infection. However, evaluating the in vivo activity of this extract is necessary too.

Dendrobium orchid polysaccharide extract: Preparation, characterization and in vivo skin hydrating efficacy

Mayuree Kanlayavattanakul, Tamonpatch Pawakongbun and Nattaya Lourith — 2019/11/12 0:00:00

Objective: Dendrobium spp., the major globally commercializing tropical orchid, has been continuously used in Chinese medicine. However, preparation and standardization of this therapeutic orchid including its clinical evidence for topical application are sparely to be exploited. Methods: Maceration of the white orchid (Dendrobium cv. Khao Sanan) in water was varied on the extraction time. The extraction yields and total polysaccharide content of the extracts were compared. The best extract condition was selected, standardized, and evaluated on safety and skin hydrating efficacy in human volunteers. Results: The extraction for 3 h gave the extract with polysaccharide content that significantly (P < 0.001) greater than the other extraction times with the following specification: yield [(35.60 ± 2.51)%], moisture content [(7.65 ± 0.21)%], total polysaccharide, sugar, reducing sugar and ash content [(72.95 ± 2.37)%, (34.38 ± 0.00)%, (16.28 ± 0.00)%, and (0.12 ± 0.00)%], pH (4.42 ± 0.01), viscosity [(15.07 ± 0.12) cps] swelling and water absorption capacities [(10.00 ± 0.00)%, (1.19 ± 0.24) g/g]. This antioxidative polysaccharide caused no skin irritation with a better skin hydrating efficacy than the untreated skin and the benchmark as examined in 22 Thai volunteers. Conclusion: White Dendrobium is ready to be supplied as a specialty ingredient with a safe and efficient profile for skin dryness therapy. This innovative application of the identified traditional Chinese medicinal herb flows in the main stream of the consumers’ preferences and demand upon natural derived products.

Monosaccharide analysis and fingerprinting identification of polysaccharides from Poria cocos and Polyporus umbellatus by HPLC combined with chemometrics methods

Jie Liu, Jing Zhou, Qian-qian Zhang and Min-hang Zhu — 2019/11/12 9:40:23

Objective: Poria cocos and Polyporus umbellatus are similar medicinal fungi in traditional Chinese medicines. A method for fingerprint analysis of monosaccharide composition of polysaccharides by HPLC combined with chemometrics methods has been developed for characterization and discrimination of them in this research. Methods: The polysaccharides were extracted by decocting in water, and then completely hydrolyzed with hydrochloride. Monosaccharides in the hydrolyzates were derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP) for HPLC analysis. More than 20 batches of P. cocos and P. umbellatus from different regions were analyzed. Results: The fingerprints of P. cocos showed five common characteristic peaks, which were identified by comparing with the reference substances. The five peaks corresponded to the derivatives of mannose, ribose, glucose, galactose, and fucose. At the same time, the fingerprints of P. umbellatus showed eight common characteristic peaks, of which seven were identified as the derivatives of mannose, ribose, rhamnose, glucose, galactose, xylose, and fucose. Moreover, the similarity of their fingerprints was respectively calculated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM published by China Pharmacopoeia Committee (Version 2004A). And the data were further processed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). The similarity evaluation and HCA indicated that there were no significant difference in P. cocos or P. umbellatus samples from different geographical regions, but PCA was performed to characterize the difference in monosaccharide constituents between P. cocos and P. umbellatus, and linear discriminant analysis (LDA) showed the overall correct classification rate was 100%. Conclusion: The fingerprint analysis method of monosaccharide composition of water-soluble polysaccharides can distinguish Poria cocos and Polyporus umbellatus, and can be applied for the authentication or quality control for Poria cocos and Polyporus umbellatus.

Synergistic effect of α-mangostin on antibacterial activity of tetracycline, erythromycin, and clindamycin against acne involved bacteria

Md Iftekhar Ahmad, James E Keach, Tapan Behl and Pharkphoom Panichayupakaranant — 2019/5/7 0:00:00

Objective To evaluate the synergistic effect of α-mangostin with tetracycline, erythromycin, and clindamycin against bacteria involved in acne production. Methods A broth microdilution method was used to determine the minimum inhibitory concentration (MIC) of α-mangostin and a range of antibiotics. Synergistic effects on antibacterial activity were determined based on their own MIC, and then by using a checkerboard method and a time-kill assay at 37 °C for 24 h. Results α-Mangostin exhibited antibacterial activity against Propionibacterium acnes, Staphylococcus aureus, S. epidermidis and S. pyogenes with MIC values of 0.78, 3.13, 0.78, and 6.25 μg/mL, respectively. The results of the checkerboard assay showed that α-mangostin produced synergistic effects with tetracycline, erythromycin and clindamycin against all tested bacteria, with a fractional inhibitory concentration index (FICI) between 0.09 and 0.32. Moreover, time-kill curve data indicated that α-mangostin increased the antibacterial activity of tetracycline, erythromycin and clindamycin. Conclusion These findings suggested that α-mangostin may be used to enhance the antibacterial activity of some antibiotics against bacteria involved in acne production.

Boost anti-oxidant activity of yogurt with extract and hydrolysate of cinnamon residues

2019/5/7 15:22:54

Objective: This study was conducted to explore the potential use of cinnamon residues (twigs and leaves) in boosting the anti-oxidant activity of yogurt. Methods: The cinnamon bark was used as the benchmark. The extracts of cinnamon bark (BW), twigs residue (TW), and leaves residue (LW) were prepared by using water, whereas the hydrolysates of cinnamon bark (BE), twigs residue (TE) and leaves residue (LE) were prepared via cellulase hydrolysis. The extracts and hydrolysates were then co-fermented respectively with the skimmed milk to produce yogurt. Results: Results obtained indicated that BW and TE yogurt possessed the highest anti-oxidant activity. In vitro digestion improved the anti-oxidant activity of yogurt significantly (P < 0.05). DPPH activity of the LW yogurt was improved drastically after in vitro digestion. Although the total phenolic content (TPC) and total flavonoids content (TFC) of LW were lower than BW, the anti-oxidant activity of LW yogurt was not significantly different (P < 0.05) with the BW yogurt after digestion. Conclusion: This study suggested that the anti-oxidant activity of the cinnamon yogurt was influenced by complex protein-phenolic interactions.

Chemical constituents from heartwoods of Caesalpinia sappan with antiplatelet aggregation activities

2019/5/7 15:22:54

Objective To clarify the active constituents of the heartwoods of Caesalpinia sappan, a traditional Chinese medicine with the functions of promoting blood circulation (Huoxue in Chinese) and removing blood stasis (Quyu in Chinese). Methods The chemical constituents were isolated and purified by combination of silica gel and Sephadex LH-20 column chromatography, along with semipreparative HPLC. Their chemical structures were established by multiple spectroscopic methods and comparison with literature data. The in vitro antiplatelet aggregation activities were evaluated using mouse platelet induced by AYPGKF-NH2, a gold agonist of protease-activated receptor 4 (PAR4). Results Two new phenols, methyl 2-(4,4′,5′-trihydroxy-2′-(methoxymethyl) biphenyl-2-yloxy) acetate (1) and 1′-methylcaesalpin J (2), together with 24 known compounds (3-26), were isolated from the heartwoods of C. sappan. Among them, sappanchalcone (16) and brazilin (20) showed inhibitory activities against mouse platelet aggregation with IC50 values of 114.8 μmol/L and 100.8 μmol/L, respectively. Conclusion Antiplatelet compounds from C. sappan targeting at PAR4 are reported for the first time.

In vivo investigation of antimicrobial effects of cinnamaldehyde using immunosuppressed ICR mice with invasive pulmonary aspergillosis

2019/5/7 0:00:00

Objective: Growing problem of antibiotic resistance in Helicobacter pylori, as a common cause of chronic gastritis and even stomach cancer, demands searching for novel candidates of herbal sources. This study is aimed at assessing the antimicrobial activity of aqueous extract obtained from Quercus brantii var. persica seed coat (Testa) on H. pylori isolated from gastric biopsy specimens. Methods: Such specimens were collected from 100 patients presenting with endoscopic gastroduodenal findings. Testa extracts were prepared from Persian Oak forests in the province of Kohgiluyeh and Boyer-Ahmad, IRAN. H. pylori isolates were obtained by a series of standard bacteriology tests and cell culture, then were confirmed by PCR. The activity of testa extracts towards 25 H. pylori isolates was assessed by well diffusion method, microdilution assay, and a disk diffusion assay in vitro. Results were analyzed statistically by one-way ANOVA analysis. Results: Aqueous extract of testa demonstrated an antimicrobial activity with zone diameters of inhibition ranged from 0 mm to 40 mm. Its inhibitory activity increased simultaneously with increasing extract concentration. The lowest MIC and MBC were both recorded as 2 μg/mL. Anti-H. pylori activity of testa extract was approximately close to tetracycline and metronidazole and less than amoxicillin. A potent extract of testa possessed significant inhibitory activity (P < 0.05). Conclusion: Testa extract is suggested as a natural therapeutic source against the gastric H. pylori infection. However, evaluating the in vivo activity of this extract is necessary too.

Bioactivity of methanolic extract of Brassica juncea in animal model of diabetes mellitus

Kumar Anuj, Rana Akhilesh, Singh Amit and Singh Alok — 2019/5/7 15:22:54

Objective To study the bioactivity of methanolic extract of Brassica junceaon animal model of diabetes mellitus along with its effect on diabetic and metabolic parameters. Methods Diabetesmellitus was induced in rats by injecting streptozotocin (60mg/kg) intraperitonealy. Blood glucose was measured on day 3by GOD-POD method to confirm the diabetes mellitus. Rats having fasting blood glucose >250 mg/dLwere further selected for study and they were divided into four groups, control, control+streptozotocin, streptozotocin+ metformin(75mg/kg) and streptozotocin+ extract of B. juncea(450mg/kg). Each group consistedof six rats of either sex. Metformin and experimental extract were administered for 21 d. Triglyceride, cholesterol level were measured on day 21 by commercially available kit. Blood glucose was measured on days 7 and 21. Anti-oxidant potential was assessed by estimating extent of lipid peroxidation (LPO) by malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD) and glutathione (GSH) in liver, kidney, pancreas, muscle tissues on day 21. Unpaired and paired student’s t-test were applied for statistical analysis. Results The extract of B. junceashowed significant decrease in blood glucose level on day 21. The treatment group showed significant difference in oxidative stress by increasing SOD and GSH and decreasing LPO and NO activity on day 21. The treatment didn’t show statistically significant difference of cholesterol, and triglycerides level on day 21. Conclusion The study showed anti-hyperglycemic and anti-oxidative properties of methanolic extract of B.juncea.

Ginsenoside Rd improves behavioral impairment of rats with acute plateau status by modulating synaptic plasticity

2019/11/12 9:40:23

Objective: To investigate the protective effect of ginsenoside Rd on the improvement of the behavior and synaptic plasticity in rats with acute plateau status. Methods: A total of 60 Wistar rats were randomly divided into the control group, the model group, and the intervention group, with 20 rats in each group. The model was established in low-pressure oxygen chamber simulating the plateau, and the intervention group was administered with ginsenoside. Electron microscope was used to observe synaptic ultrastructure of hippocampal CA1 area, and analyze the structural parameters on the Gray I synaptic interface. Morris water maze and Y electric maze experiment were used for behavioral detection. Results Compared with the control group, the number of electrical stimulation required for rat to avoid was increased in the model group, the latency in the Morris water maze was prolonged, the swimming distance was increased, and the frequency of crossing the platform was decreased. Under the electron microscope, the synaptic cleft was increased, the length of the synaptic active area was shorter, the postsynaptic density (PSD) was thinner, the flat synapse was increased, and the concave and perforated types were significantly reduced. Compared with the model group, the number of electrical stimulation required for rat to avoid was decreased in the intervention group, the latency in the Morris water maze was shortened, the swimming distance was decreased, and the frequency of crossing the platform was increased. Under the electron microscope, the synaptic cleft was decreased, PSD was thicker, the flat synapse was decreased, and the concave and perforated types were increased. Conclusion Low pressure and low oxygen environment of plateau damages the plasticity changes of the synaptic structure and function. And to a certain extent, ginsenoside Rd reverses Gray I synaptic interface structure parameters, so as to improve the behavior performance of model rats at high altitude condition.

Towards a scientific rationale for traditional properties of Chinese medicinal plants: a€?naturesa€? and a€?flavorsa€?

2019/7/31 15:54:06

Objective According to theory of Chinese medicine property, the ecological environment shapes properties (natures and flavors) of Chinese materia medica (CMM) and there are close relationships between certain natures and flavors. However, to date these observations have not been validated scientifically in the context of the whole flora of a region. The present study aims to address this gap. Methods We collected geographical distributions of 3637 vascular plant species native to the Xinjiang region of northwest China, of which around 812 are medicinal plants. The CMM property characters of these medicinal plants were also collected. All medicinal plants were then analyzed in the occurrence of their natures (cold, cool, neutral, warm, and hot) and flavors (sweet, bitter, sour, salty, and pungent). Possible correlations between these properties and environmental factors, notably climate, as well as correlations with plant species richness of the wider native flora of Xinjiang were then investigated using permutation test and regression. Results Cold species (38.5%) were more numerous than warm (21.0%) and neutral (22.3%), while bitter species (43.3%) were more numerous than pungent (25.6%), sweet (22.4%) and sour (6.7%) ones. Species with bitter flavor were most commonly also cold (48.6%) in nature, sour flavor usually coincided with cold (44.2%) or cool nature (26.0%), whereas pungent or sweet species tended to be warm (34.4%, 28.4%) or neutral (27.8%, 28.4%). The percentages of species of cold/cool nature, bitter flavor were higher in mountainous regions, and the percentages of warm/neutral and pungent/sweet/sour were higher in basins. The percentages of species of bitter, cold/cool, bitter were positively correlated with actual evapotranspiration (AET), plant species richness andbut had a bell-shaped relationship with potential evapotranspiration (PET), whereas the inverse situation was shown for warm/neutral and pungent/sweet/sour species. Conclusion Analysis of the data in Xinjiang region of China supports the qualitative observations concerning natures and flavors in traditional theory of Chinese medicine property. Specifically, (i) certain natures and flavors co-occur more frequently than if randomly distributed; (ii) correlations between proportions of natures and flavors and climatic variables suggest a role for the environment in shaping the properties of CMM.

Parasitic relationship of Cistanche deserticola and host-plant Haloxylon ammodendron based on genetic variation of host

2019/7/31 15:54:06

Objective Cistanche deserticola is a famous and endangered medicinal plant that is parasitic upon Haloxylon ammodendron with rather low parasitic rates. It is important to find high affinity germplasms for increasing the survival of C. deserticola. However, little is known in genetic variation and high affinity populations of H. ammodendron in China. Methods In this study, 98 accessions of H. ammodendron seeds were collected from five regions covering almost the entire natural distribution of H. ammodendron in China. Their genetic variations were analyzed using AFLP and ITS by the maximum parsimony method, and a dendrogram was constructed using the unweighted pair-group method with arithmetic average (UPGMA). The parasitic rates of C. deserticola on different accessions of H. ammodendron were calculated in the field experiment. Results Both AFLP and ITS methods consistently revealed that there was a high level of genetic diversity in the natural populations of H. ammodendron. Hierarchical population structure analysis uncovered a clear pattern that all populations were grouped into three main clusters, and eight populations from eastern region were genetically clustered together. These regions were significantly differentiated (P < 0.05), 13.10% of variation occurred among populations, and 86.90% within populations was revealed by analysis of molecular variance (AMOVA). The populations of Inner Mongolia had the highest parasitic rates followed by Ganjiahu Reserve and Yongning Plantation for the top three, which were not completely related to the genetic variation. Conclusion Genetic characteristics of H. ammodendron in China were clarified and the order of affinity of different populations was given, which were primers for discovering high affinity germplasms.

Identification of flower herbs in Chinese pharmacopoeia based on DNA barcoding

Xue-min Wei, Xiao-yue Wang, Zi-tong Gao, Pei Cao and Jian-ping Han — 2019/7/31 15:54:06

Objective Flower herbs are an important category of traditional Chinese medicinal materials, some of which are used as healthcare tea in folk. However, the increasing adulteration of medicinal herbs is threatening consumer safety. The adulteration of flower herbs and their healthcare tea products in the market were investigated. Methods A total of 33 flower herb samples from several retail pharmacies in China were randomly collected and 27 flower healthcare tea samples were purchased online. They were identified using ITS2-based Traditional Chinese Medicine Database (TCMD). Additionally, Lonicerae Japonicae Flos and the adulterants were compared in the ITS2 secondary structures. Results There were one adulterant (Inulae Flos) in flower herb materials and eight adulterants in healthcare tea samples. Inula linariifolia was an adulterate species of Inulae Flos, Robinia pseudoacacia was of Sophorae Flos, and Lonicera macranthoides was of Lonicerae Japonicae Flos. Sophorae Flos and Lonicerae Japonicae Flos were two healthcare tea products with high adulteration rates. Conclusion The TCMD is powerful tool to identify flower herbs and the adulterants that frequently occurred in the flower herb market, especially online shops.

Mechanisms of Chinese medical formula Fangji Huangqi Decoction as an effective treatment of nephrotic syndrome based on systems pharmacology

Xue-min Wei, Xiao-yue Wang, Zi-tong Gao, Pei Cao and Jian-ping Han — 2019/7/31 15:54:06

Objective: Flower herbs are an important category of traditional Chinese medicinal materials, some of which are used as healthcare tea in folk. However, the increasing adulteration of medicinal herbs is threatening consumer safety. The adulteration of flower herbs and their healthcare tea products in the market were investigated. Methods: A total of 33 flower herb samples from several retail pharmacies in China were randomly collected and 27 flower healthcare tea samples were purchased online. They were identified using ITS2-based Traditional Chinese Medicine Database (TCMD). Additionally, Lonicerae Japonicae Flos and the adulterants were compared in the ITS2 secondary structures. Results: There were one adulterant (Inulae Flos) in flower herb materials and eight adulterants in healthcare tea samples. Inula linariifolia was an adulterate species of Inulae Flos, Robinia pseudoacacia was of Sophorae Flos, and Lonicera macranthoides was of Lonicerae Japonicae Flos. Sophorae Flos and Lonicerae Japonicae Flos were two healthcare tea products with high adulteration rates. Conclusion: The TCMD is powerful tool to identify flower herbs and the adulterants that frequently occurred in the flower herb market, especially online shops.

Berberine reverses LPS-induced repression of CYP7A1 through an anti-inflammatory effect

2019/7/31 15:54:06

In vivo antiplasmodial potential of three herbal methanolic extracts in mice infected with Plasmodium berghei NK65

2019/7/31 15:54:06

Objective The present study deals with the investigation of antiplasmodial potential of leaf methanolic extract of Aegle marmelos, Aristolochia indica and Cassia auriculata against Plasmodium berghei (NK65) infected mice. Methods The chloroquine-sensitive parasites P. berghei (1?×?106) were inoculated into Swiss albino mice intraperitoneally. The methanol extracts of three herbal plants were orally administered in P. berghei infected mice which were further assessed using the 4 d suppressive test at different doses of 150, 300 and 600?mg/kg per d. Chloroquine (CQ) was used as the standard drug with of 1.25, 2.5 and 5?mg/kg concentrations and was orally administered. Results The leaves of A. marmelos, A. indica, and C. auriculata were found to suppress P. berghei parasitaemia in Swiss albino mice by (67.0 0?±?4.02) %, (72.0 0?± 8.44)% and (52.7 7?±?2.06)% at 600?mg/kg/d with ED50 values of 284.73, 233.77 and 562.48?mg/kg, respectively. These herbal plants increased the mean survival time of infected mice and prevented body weight loss. GC-MS analysis revealed the presence of hentriacontan-16-one (C31H62O) in A. indica extract. The histopathology study showed non-toxic to kidney and liver at 600?mg/kg/body weight. Conclusions Overall results revealed that herbal plants may be active in the development of novel and cheap antimalarial compounds.

Effects of Raphani Semen on anti-fatigue and pharmacokinetics of Panax ginseng

Jian-bo?Chen, Mei-jia?Li, Li-xue?Chen and Yin-shi?Sun — 2019/7/31 15:54:06

Objective To explain the phenomenon that Panax ginseng is not compatible with Raphani Semen based on pharmacodynamics and pharmacokinetics. Methods The forced swimming time and biochemical parameters such as blood lactate (BLA), serum urea nitrogen (SUN), and hepatic glycogen (GLU) were determined for anti-fatigue experiment. The UPLC-MS/MS was used to analyze the pharmacokinetics of Rg1, Re, Rb1, and Rd after orally administration of P. ginseng and P. ginseng combined with Raphani Semen to rats. Pharmacokinetic differences of four ginsenosides between single uses of P. ginseng and combined with Raphani Semen were investigated. Results The results showed that Raphani Semen tended to significantly reduce the anti-fatigue activity of P. ginseng. Co-administration of P. ginseng and Raphani Semen had significant effects on the pharmacokinetics of the four ginsenosides in rats compared to that observed with P. ginseng extract alone. The AUC0–12?h values of the four ginsenosides in PG group were higher than the corresponding values in the PR group. It can be inferred that Raphani Semen decreased the blood exposure of the four ginsenosides in rats when it combined with P. ginseng. Conclusion The anti-fatigue activity and pharmacokinetic results showed that Raphani Semen may reduce the pharmacological actions of P. ginseng.

Comparative pharmacokinetics of naringin and neohesperidin after oral administration of flavonoid glycosides from Aurantii Fructus Immaturus in normal and gastrointestinal motility disorders mice

2019/7/31 15:54:06

Objective: To compare the pharmacokinetics of naringin and neohesperidin after oral administration of Zhishi total flavonoid glycosides (ZSTFG) extracted from Aurantii Fructus Immaturus in normal and gastrointestinal motility disorders (GMD) mice. Methods: ZSTFG was orally given to normal and GMD mice induced by atropine or dopamine. The plasma samples were incubated with β-glucuronidase/sulfatase, the total (free?+?conjugated) naringenin and hesperitin were extracted with acetonitrile. The validated HPLC-MS/MS method was successfully applied to the pharmacokinetic study. Results: The results showed that, compared with the normal group, AUC0–∞, AUC0–t and Cmax for total naringenin and hesperitin were significantly higher (P < 0.01 or P < 0.05), while CLZ/F for total naringenin and hesperitin was significantly lower (P < 0.01) in the GMD group. Tmax, t1/2z, MRT0-t, and MRT0-∞ for naringenin were longer (P < 0.01) in the GMD group than those in the normal group. Conclusion: The results showed that there were significant differences in pharmacokinetic parameters of naringenin and hesperitin between normal and GMD groups. It was suggested that the absorption of naringenin and hesperitin was increased, and the elimination processes of naringenin and hesperitin were slower in the GMD group than the normal group. The data are of value for further pharmacological studies of ZSTFG and would be useful to provide a reference for improving the therapeutic regimen of ZSTFG in clinical trials.

Synergistic activity of lawsone methyl ether in combination with some antibiotics and artocarpin against methicillin-resistant Staphylococcus aureus, Candida albicans, and Trychophyton rubrum

2019/7/31 15:54:06

Antiobesity and antihyperlipodemic effect of Ixora coccinea on Triton X-100 induced hyperlipidemia in rats: An approach to evaluate asymmetrical temperature distribution analysis using thermography

2019/7/31 15:54:06

Objective: Obesity and hyperlipidemia is the major cause of many pathological diseases with an increase side effects using allopathic drugs. The present study focuses on the effect of Ixora coccinea on Triton X-100 induced hyperlipidemia in rats and associated complications. Methodology: In vitro radical scavenging activity of I. coccinea was assessed using DPPH, FRAP and hydrogen peroxide. In vivo antiobesity and antihyperlipidimic activity of I. coccinea was tested in Triton X-100 induced hyperlipidemic rats and assessed for its biochemical parameters in blood and tissue samples. The relationship between physiological responses and regulation of body temperature was investigated by using animal surface temperature images captured with infrared camera. Results: The results of mineral analysis, antioxidant, total flavonoid and phenolic content represented high amount of mineral and had the potential to scavenge free radicals tested with DPPH, FRAP and hydrogen peroxide radicals with dose dependent activity. The highest activity was observed in aqueous extract, DPPH with 71.5% inhibition, FRAP with 56.8%, H2O2 with 33% activity at 100?μg/mL concentration. Triton X-100 induced hyperlipidemic rats when treated with I. coccinea aqueous extract showed significant activity by regulating the biochemical parameters and maintaining the lipid profile by decreasing TC, LDL-C, VLDL-C, TG and improving HDL-C levels. Similarly, the elevated levels of creatinine, urea, uric acid, AST, ALT, ALP due to induction of hyperlipidemia, were brought back to near normal levels after treatment with I. coccinea. The levels of tissue anti-oxidants enzymes like SOD and CAT were also found to be improved in treated I. coccinea groups. The whole body asymmetrical temperature distribution analysis showed that significant decreases in temperature was observed in obesity induced groups but a gradual increase in temperature (2% – –5%) was observed after treatment. Conclusion: Thus, the results indicated that I. coccinea can be a drug of choice to decrease the risk of complications associated with hyperlipidemia and obesity.

A guinea pig model of Ciwujia Injection-induced anaphylaxis for allergic substance screening

2019/7/31 15:54:07

Objective: Though especially efficient for cardiovascular and cerebrovascular diseases treatment, many serious anaphylactic diseases could be induced by Ciwujia Injection (CWJI). However, study of the mechanism and detection of allergies have been investigated by the unknown sources of allergenic substances. In this study, a guinea pig model which could mimic the symptoms of anaphylactic reactions induced by Ciwujia Injection (CWJI) was modeled and used to screen the allergenic substance of CWJI. Methods: Guinea pigs were sensitized three times every other day with CWJI and excitated 14 d after the last sensitization administration. Then, the histamine, trypsin, IL-4 and IFN-γ levels, the Annexin V positive rate of peritoneal mast cells (PMC) were detected, the numbers of B lymphocyte and the pathological changes were also analyzed to verify the guinea pig allergy model, PCA test and IgE antibody levels were determined to study the mechanism. Results: The levels of total IgE, histamine, and trypsin were significantly increased after CWJI sensitization, IL-4 level was elevated, Annexin V positive of PMC cell rate, local skin reactions, and declined IFN-γ were observed after excitation. Histological examination showed that mild pathological changes in lungs were found. Conclusion: This guinea pig model may provide a powerful tool to study the mechanism in CWJI induced anaphylaxis and screen the allergic source of CWJI.

Formulation optimization, in situ intestinal absorption and permeability of psoralen and isopsoralen

2019/7/31 15:54:07

Objective To optimize a self-emulsifying drug delivery system (SEDDS) formulation for psoralen and isopsoralen (PSO and IPSO) isolated from Psoraleae Fructus. Methods A D-optimal design was used to investigate the influence of oil percentage, surfactant percentage and cosurfactant percentage on several properties of SEDDS including particle size, polydispersity, equilibrium solubility, in situ intestine absorption rate and intestinal permeability. Furthermore, the desirability function approach was applied to obtain the optimal formulation for the system. Results The oil percentage, surfactant percentage and cosurfactant percentage were optimized to be 53.6%, 35.7% and 10.7%, respectively, which means the model is available. Conclusions The D-optimal design is valuable to optimize the SEDDS formulation and understand formulation compositions’ functions on SEDDS properties.

Quinoline alkaloids isolated from Scolopendra subspinipes mutilans

2019/7/31 15:54:07

Objective To study the quinoline alkaloids from the ethanol extract of Scolopendra subspinipes mutilans (SSM). Methods The chemical constituents were isolated and purified by macroporous resin column, medium pressure preparation chromatography, and semi-preparative HPLC. Their structures were elucidated by IR, MS, and NMR experiments. Results Three quinolone alkaloids were obtained and identified as 3-hydroxy-4-methoxyquinolin-8-yl hydrogen sulfate (1), jineol-8-sulfate (2), and jineol (3), respectively. Conclusion Compound 1 is a new compound from SSM.

Quantitative and HPLC fingerprint analysis combined with chemometrics for quality evaluation of Codonopsis Radix processed with different methods

2019/5/7 0:00:00

Objective: To observe the effect of different processing methods on the quality of Codonopsis Radix and provide data reference for the reasonable processing in the producing areas. Methods: An analytical strategy that combined quantitative and HPLC ?ngerprint analysis with chemometrics was developed for the quality evaluation of Codonopsis Radix. Lobetyolin, polysaccharide, sucrose, glucose, and fructose were detected simultaneously in Codonopsis Radix samples treated with different processing methods including dryer-drying, sun-drying, shade-drying, sulfur fumigation and kneading, etc. Results: The results showed that different processing methods had an obvious impact on the quality of Codonopsis Radix. Sun-drying or shade-drying was recommended, and sulfur fumigation should be avoided. And dryer-drying at 40 °C ? 50 °C was suggested to meet the requirement for large scale processing. In addition, based on the content of polysaccharide, glucose, and fructose, it was advised to knead for three times. As for lobetyolin and sucrose, kneading should be avoided. Conclusions: Our ?ndings demonstrated that processing method had a big impact on the quality of Codonopsis Radix. Based on the results, the appropriate processing methods directed at different requirements were recommended. The study could lay a foundation for the reasonable processing of Codonopsis Radix in producing areas.

Effect of Marsdenia tenacissima extract on G2/M cell cycle arrest by upregulating 14-3-3σ and downregulating c-myc in vitro and in vivo

2019/5/7 0:00:00

Objective: Marsdenia tenacissima extract (MTE) is a traditional Chinese herbal medicine with anti-cancer activity. In some previous studies, different mechanism actions of the anti-cancer effect of MTE have been revealed. In this study, we first observed that MTE exhibits G2/M cell cycle arrest on two different human breast cancer cell lines, MDA-MB-231 and MCF-7 by mediating 14-3-3σ and c-myc. Methods: The effect of MTE on G2/M cell cycle arrest was evaluated in MDA-MB-231 and MCF-7 cell lines. MTT assay was done for evaluation of cell viability, Flow cytometry was employed for cell cycle analysis, Western blotting analysis and immunohistochemistry were performed to analyze the expression of G2/M cell cycle-related key protein in cells and tissue samples. Animal studies have been conducted to elucidate the anti-tumor effect of MTE. Results: Cell cycle is the backbone for developing cancer. Cell cycle proteins play a major role in the progression of cell cycle and cell proliferation. However, some key proteins directly or indirectly modulate the action of cell cycle protein that highly affect cell cycle regulation. In order to investigate cellular proliferation of cancer, we observed that MTE induced the upregulation of 14-3-3σ and downregulation of c-myc, and then reduced the expression of G2/M cell cycle associated key protein, leading to the inhibition of cellular entry into mitosis phase. We also confirmed that MTE exerted a significant antitumor effect on the MDA-MB-231 xenograft model in vivo. Conclusion: G2/M cell cycle arrest occurred by the action of MTE, mediated by the upregulation of 14-3-3σ as well as downregulation of c-myc in MDA-MB-231 and MCF-7 cell lines.

Effects and mechanism of total phenols of Magnolia officinalis combined with Maijunan Tablets on blood pressure of spontaneous hypertensive rats

Ming-li Zhu, Wei Wang, Feng Qin and Yong Chen* — 2019/5/7 0:00:00

Objective: Maijunan (MJA) Tablets is a protected variety of traditional Chinese medicine (TCM) consisted of Pueraria lobata, hydrochlorothiazide (HTCZ), Uncaria rhynchophylla (366:1:980) and excipient. In the present work, MJA was consisted of the total flavones of P. lobata, HCTZ and total alkaloids of U. rhynchophylla (40:11:75). The combination of MJA and the total phenols of Magnolia officinalis (M-MJA) was consisted of the total flavones of P. lobata, the total phenols of M. officinalis, HCTZ and the total alkaloids of U. rhynchophylla (40:40:11:75). The aim of this work was to examine the effect and mechanism of M-MJA on the blood pressure of spontaneous hypertensive rats (SHRs). Methods: Adult male SHRs were randomly divided into control group, MJA group (180 mg/kg·d), and the M-MJA group (218 mg/kg·d) (n = 5). SHRs were orally administered with M-MJA and MJA respectively once a day for 8 weeks, the blood pressure of SHRs was measured every two weeks, and the biochemical indicators related to blood pressures were detected at the last dosing. Results: After oral administration of M-MJA to SHRs once a day for 8 weeks, the systolic and diastolic blood pressures of SHRs were deceased significantly. M-MJA affected renin-angiotensin-aldosterone system by decreasing the levels of Ren, Ang II and ALD, affected the endothelial function by decreasing the levels of ET-1 and 20-HETE, and increasing the level of eNOS, affected the oxidative stress by increasing the protein expression of Nrf2 and the activities of HO-1 and GSH-Px, and decreasing the protein expression of CYP2E1 and CYP4A, as well as the content of MDA. Conclusion: These results indicated that M-MJA could regulate the renin-angiotensin-aldosterone system, improve endothelial function, and inhibit CYP4A activity to reduce the production of 20-HETE, alleviate the oxidative stress disorder of the visceral organs, and eventually exert antihypertensive effect. Additionally, the anti-oxidant ability, regulating the renin-angiotensin-aldosterone system and improving endothelial function of M-MJA are more powerful than that of MJA, suggesting that M-MJA may have a better anti-hypertensive effect than MJA.

Metabolic profiling of icaritin in rats using UHPLC-Q/TOF-MS

2019/5/7 15:22:54

Objective: To identify the in vivo metabolites of icaritin and speculate its metabolic profiling in rats. Methods: The plasma, bile, urine, and feces of rats were collected after orally administration of icaritin at a dose of 100 mg/kg and detected by an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS/MS) in both positive and negative modes. The data of treated and control groups were compared and analyzed with the aid of Metabolynx XS software. Results: A total of 25 metabolites were identified in the biosamples, and 14 of them were reported for the first time to our knowledge. Conclusion: The main metabolite types of icaritin in rats were glucuronide conjugation, methylation, hydroxylation, reduction, and acetylation.

Over-expressing root-specific β-amyrin synthase gene increases glycyrrhizic acid content in hairy roots of Glycyrrhiza uralensis

Mo-li Hua and Yun-hui Xu — 2019/5/7 0:00:00

Objective: Glycyrrhizia uralensis, one of the most widely-used traditional Chinese medicines, is mainly cropped in China. However, many cultivars are less in glycyrrhizic acid than Chinese Pharmacopoeia requires. In this paper, we improved glycyrrhizic acid by regulating β-amyrin synthase gene (GuBAS). Methods: Tobacco root-specific promoter TobRB7 and GuBAS cDNA were obtained and combined with linearized pCAMBIA1305.1 to construct root-specific plant expression vector which was later transformed into Agrobacterium rhizogenes ACCC10060 by electrotransformation. The cotyledons and hypocotyls of G. uralensis were infected by the recombinant A. rhizogenes ACCC10060 to induce hairy roots. The GA content was quantified by HPLC. Results: The PCR and sequencing results both showed that three transgenic hairy root lines were obtained. The copy number of GuBAS in these transgenic hairy roots was intended by qRT-PCR to be 3, 7, and 4. GA was detected in the three transgenic hairy roots but in wild hairy roots. Conclusion: Over-expressing GuBAS root-specifically in hairy roots of G. uralensis enhanced GA accumulation.

Simultaneous determination of 15 flavonoids from different parts of Scutellaria baicalensis and its chemometrics analysis

2019/1/14 0:00:00

Objective The aerial parts of Scutellaria baicalensis were used as Huangqin Tea for thousands of years and mainly contain flavonoids which contribute to its bioactivities. However, there is no appropriate quality evaluation method of Huangqin Tea, and three flavanones of isocarthamidin-7-O-β-D-glucuronide, carthamidin-7-O-β-D-glucuronide, and isoscutellarein-8-O-β-D-glucuronide with high contents in the aerial parts have never been defined quantitatively. Here, an HPLC-DAD method for simultaneous determination of 15 flavonoids and systematically compared their contents and distribution in the roots, stems, leaves, and flowers of S. baicalensis was established. Methods Under the HPLC-DAD chromatographic conditions, 77 batches of samples of S. baicalensis were analyzed. Meanwhile, the chromatographic fingerprint of different parts of S. baicalensis was established. Subsequently, principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (OPLS-DA), and clustering heat map were performed based on the contents of 15 flavonoids in different parts of S. baicalensis. Results The results showed significant differences in the contents and distributions of 15 flavonoids among the different parts of S. baicalensis. The chemical composition of stems showed some similarities to leaves, and their contents were all lower than leaves. The contents of isocarthamidin-7-O-β-D-glucuronide [(106.66 ± 22.68) mg/g], carthamidin-7-O-β-D-glucuronide [(19.82 ± 11.17) mg/g], and isoscutellarein-8-O-β-D-glucuronide [(3.10 ± 1.73) mg/g] were the highest in leaves. The content of apigenin-7-O-β-D-glucopyranoside and chrysin-7-O-β-D-glucuronide were the highest in flowers. The contents of baicalin, baicalein, wogonoside, wogonin, alpinetin, and oroxylin A were higher in roots than in other parts. Conclusion The method was fully validated and could be effectively used to characterize the contents and distributions of main flavonoids in the different parts of S. baicalensis. It may lay a foundation to establish the quality evaluation system for Huangqin Tea.

Pharmacokinetic and NMR metabolomics approach to evaluate therapeutic effect of berberine and Coptidis Rhizoma for sepsis

2019/1/14 0:00:00

Objective: Sepsis, a systemic response to infection, often leads to end-organ dysfunction. Despite its high rates of mortality and morbidity, its pathophysiology is still poorly understood. Coptidis Rhizoma and its main active alkaloid compound, berberine, have been as anti-bacterial and anti-inflammatory drugs used in clinic. The objective of this study was to gain more insights towards understanding the sepsis associated with drug absorption and disposition and treatments of berberine and Coptidis Rhizoma dynamically. Methods: Pharmacokinetic and metabolomic studies of Coptidis Rhizoma and its main active component berberine have been performed. Results: Cecal ligation and puncture (CLP) induced sepsis showed marked changes of metabolites concerning energy metabolism and amino acids metabolisms, which could be reversed towards the normal state by Coptidis Rhizoma and berberine. Conclusion: Berberine exhibited an equivalent and even better therapeutic effect than Coptidis Rhizoma.

Positive effects of low dose IMPX977 on Rett syndrome related MeCP2 targeted-genes

2019/1/14 0:00:00

Objective To investigate the effect of IMPX977 on MeCP2 targeted-genes and the feasibility of IMPX977 acting as a therapeutic candidate drug for Rett syndrome by genomewide transcription profiling. Methods Rats’ cortex of control group, IMPX977-treated low-dose group (10 mg/kg), and IMPX977-treated high-dose group (30 mg/kg) were collected and RNA was extracted from the tissues. Then, RNA was subjected to RNA-sequencing. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) were used in functional enrichment analysis of differentially expressed genes. Results Six MeCP2 targeted-genes were identified in the low/control categories, but not in the high/control categories. Conclusion Low-dose treatment of IMPX977 (10 mg/kg) showed a positive effect on MeCP2 targeted-genes and it may serve as a drug candidate for Rett syndrome therapy with proper dosage.

Neuroprotective effect of Sanqi Tongshuan Tablets on sequelae post-stroke in rats

2019/1/14 0:00:00

Objective To identify the therapeutic effect and possible mechanisms of Chinese medicine Sanqi Tongshuan Tablets (SQTS) on sequelae post-stroke in rats. Methods The rat cerebral ischemia sequelae post-stroke models were successfully induced by blocking the middle cerebral artery with electric coagulator after the seventh week and balance beam test ≤4. The rats were then received with SQTS (0.5, 1, and 2 g/kg) and Naodesheng (NDS, 1.24 g/kg), Vinpocetine (VP, 1.55 mg/kg) for 30 d. The beam-walking test and shuttle test were performed before and after 10, 20, and 30 d of administration. In addition, histopathology changes and GAP-43, GFAP were measured by H&E staining and immunohistochemisty. Results The model displayed signs of brain damage on motor function, learning and memory function and histopathology. After 30 d of treatment, SQTS at different dose (0.5, 1.0, and 2.0 g/kg) restored the beam-walking scores by 21.7% (P > 0.05), 30.4% (P > 0.05), and 39.1% (P < 0.05); Decreased electric shock by 35.0% (P > 0.05), 50.0% (P > 0.05), and 75.0% (P < 0.05), respectively. On the other hand, the histological changes were less severe and the GAP-43 expression increased in hippocampal CA1 and cortical region. Conclusion SQTS showed therapeutic benefits on sequelae post-stroke in rats, which might be through the pathway of regeneration or neuroplasticity.

QR code labeling system for Xueteng-related herbs based on DNA barcode

2019/1/14 0:00:00

Objective Xueteng-related herbs include Jixueteng, Daxueteng, Dianjixueteng, and other medicinal plant stems that release reddish-brown juices when cut. However, similarity in phonetic spelling and sound leads to their misuse in clinic or commercial circulation. Accurate labeling is imperative as well as effective species identification. Methods In this study, the ITS2 sequences of 76 samples of Xueteng-related herbs were obtained and analyzed to identify them. And then they were converted into QR codes using the open source PHP QR code. Also, a DNA barcode reference library was established according to these sequences and was used to authenticate the 25 samples of Xueteng-related herbs collected from the market. Results The lengths of the ITS2 sequences of different Xueteng-related herbs ranged from 207 to 235 bp and the GC contents were 57.5%?71.0%. Jixueteng, Daxueteng, and Dianjixueteng were clustered into three clades respectively in the neighbor-joining (NJ) phylogenetic tree, and the efficiency of the BLAST method was 100%. The ITS2 sequences of different Xueteng-related herbs were presented vividly and specifically in QR code. Twenty-two of all 25 commercial samples were consistent with the original labels, whereas three samples marked “Dianjixueteng” were authenticated as “Jixueteng”. Conclusion QR code labeling system based on DNA barcode is an effective labeling system of Xueteng-related herbs for their circulation regulation.

Screening potential α-glucosidase inhibitors from Anemarrhena asphodeloides using response surface methodology coupled with grey relational analysis

2019/1/14 0:00:00

Objective: To screen potential α-glucosidase inhibitors from Anemarrhena asphodeloides. Methods: Response surface methodology employing Box-Behnken design was used to optimize conditions for the extraction of α-glucosidase inhibitory active compounds from A. asphodeloides. The powders (20.0 g) of A. asphodeloides. were extracted under the optimized conditions. The extract was applied to a D-101 macroporous resin column. It was eluted with ethanol and water to give six fractions. Compounds from the active fraction were identified by UPLC-Q-TOF-MS. The structure-activity relationship was discussed based on grey relational analysis. Results: The optimum extraction conditions were as follows: ethanol concentration, 100%; extraction temperature, 51°C; and solvent to solid ratio, 23 mL/g. It indicated that the active compounds were concentrated into 80% ethanol fraction. Twenty five steroid saponins from 80% ethanol fraction were identified by UPLC-Q-TOF-MS. Peaks 19 and 23 were tentatively identified as new structures. The predicted α-glucosidase inhibitory activities of the compounds were 7 > 2 > 1 >22 >23 > 3 > 9 > 21 > 24 > 4 > 13 > 8 > 14 > 16 >17 > 25 > 6 >19. Conclusion: The fraction eluted by 80% ethanol showed the best inhibitory activity. After analyzing the data of UPLC-Q-TOF-MS, 25 steroid saponins were tentatively identified in this fraction.

Synergistic interactions of catalpol and stachyose in STZ-HFD induced diabetic mice: synergism in regulation of blood glucose, lipids, and hepatic and renal function

Xiao-fang Chen, Deng-qun Liao, Zhen-xian Qin and Xian-en Li — 2019/1/14 0:00:00

Objective Rehmanniae Radix has been traditionally used to treat diabetes. Catalpol (CAT) and stachyose (STA) are two of the main bioactive compounds in Rehmannia Radix and found to have similar therapeutic effects on diabetes and its complications. In this paper, we aimed to investigate whether there were synergistic therapeutic effects of CAT and STA on diabetes. Methods Streptozotocin (STZ) with the feeding of high-sugar-high-fat diet (HFD) was applied to induce diabetic C57BL/6 mice. STZ-HFD induced diabetic mice were then divided into model and six medical-treated groups: metformin (MET), STA, CAT, and three combinations of CAT:STA (1:1, 1:2, 2:1). Blood, liver, and kidney samples were isolated after six-week oral administration for biochemical assays of serum lipids, the indicators of kidney and liver functions and HE staining for liver tissues. Results It turned out that CAT, STA and their three combinations (1:1, 1:2, 2:1) could effectively control body weight, blood glucose, kidney weight and liver weight index and well regulate levels of TC, HDL-c, TG, ALT, and TBA. In addition, CAT and its combination with STA at the ratio of 2:1 could significantly improve albumin content, compared to that in model group. STA and CAT and their combinations showed the improvements on kidney function in terms of urinary creatinine (Ucr). However, there were no such consistent observations on serum creatinine (Scr) and creatinine clearance rate (Ccr). The combination of CAT and STA at the ratio of 1:1 exhibited the better adjusting effects on kidney weight and liver weight indexes and the levels of ALT, Ucr, Scr, and Ccr. Our results demonstrated that the combinations of CAT and STA especially 1:1 showed similar or better improvements on diabetes-associated complications, compared to the sole CAT or STA treatment. Conclusion Thus, we concluded that there were synergistic therapeutic effects between CAT and STA on STZ/HFD-induced type 2 diabetes. This project provided insights and technical supports for the innovation of discovering bioactive constituents in Rehmannia Radix and studying its integrative mechanism in curing diabetes.

Effect of bioadhesive excipients on absorption of total flavonids from Puerariae Lobatae Radix transporting across Caco-2 cell monolayer

Ying Li, Yi-qun Song and Chun-yan Zhu — 2019/1/14 0:00:00

Objective: Pueraria total flavonids (PTF) can treat cardiovascular and cerebrovascular diseases, but it has poor membrane permeability and oral bioavailability. Some excipients, such as carbomer, chitosan, and hydroxypropyl methylcellulose, can improve the oral bioavailability. Traditional in vitro evaluation techniques, including the rat intestinal perfusion and cell line models, cannot evaluate PTF absorption and holistic transporters. Methods: This study evaluated excipients’ adhesiveness and effect on PTF transport across Caco-2 cell monolayer. cDNA microarrays identified gene expression changes in Caco-2 cells exposed to PTF and PTF with excipients, and revealed the mechanism underlying the effect of excipients on PTF absorption. Results: In vitro adhesion and transport experiments across Caco-2 showed that excipients had higher adhesiveness to gastric mucosa and transport efficiency across Caco-2 cells than PTF alone. The interaction of PTF with excipients significantly changed the expression of some genes, which might influence the absorption rate of PTF. Conclusion: Different bioadhesive polymers can improve intestinal absorption of PTF, which was related to some genes affiliated to the ATP-binding cassette (ABC) and solute carrier transporter (SLC) to some extent.

Neuroprotective effect of Paeoniae Radix Rubra on hippocampal CA1

2019/1/14 0:00:00

Objective: Stroke is the second leading cause of death worldwide. This study aimed to investigate the neuroprotective effect of Paeoniae Radix Rubra (PRR) on ischemic stroke of mice. Methods: The focal ischemic stroke model was produced via middle cerebral artery occlusion. The experimental mice were divided into four groups: vehicle-sham group, PRR-sham group, vehicle-ischemia group, and PRR-treated ischemia group. The cerebral infarction volume was detected with TTC staining. The number of neurons in the hippocampal CA1 of the ischemic side, and the activation of astrocytes and microglia were observed via immunohistochemical staining. Western blotting was used to determine the expression changes of SOD1, SOD2, and Catalase protein levels in the hippocampus. Results: PRR significantly reduced the cerebral infarct volume induced by ischemic injury and inhibited the astrocytes and microglia activation in the hippocampal CA1 region. The decreased levels of SOD1, SOD2, and Catalase that was induced by ischemic reperfusion were simultaneously improved after PRR treatment. Conclusion: PRR improved neuronal injuries that were induced by transient cerebral ischemia via inhibiting gliosis and elevating antioxidants.

Monitoring unbound warfarin in drug combination therapy by pharmacokinetics and fluorospectrometry

2019/1/14 0:00:00

Objective Monitoring the unbound drug concentration in blood in combination therapy is necessary, because its concentration determines the efficacy of drug therapy. This study was designed to explore the effect of Dan Hong Injection (DHI) on the unbound warfarin using two approaches including an in vivo pharmacokinetic and in vitro fluorescence studies. Methods The effect of DHI on the pharmacokinetic properties of the unbound warfarin was investigated by a microdialysis sampling method coupled with LC-MS/MS. The effect of DHI and salvianolic acid B (SaB) on warfarin binding with bovine serum albumin (BSA) was conducted by fluorescence spectrometry. Results The AUC0-tn of warfarin with DHI group was higher than that of warfarin alone group. The result showed that DHI could increase the concentration of unbound warfarin in rat blood, which may be due to the competition between warfarin and DHI as well as some its components binding to serum albumin. The competition process was demonstrated by fluorescence study. Conclusion Combination therapy of DHI with warfarin could enhance the release profile of warfarin from serum protein.

Typha angustifolia extract reduces diet-induced hyperlipidemia in rats

Zheng Gong, An-rong Jiang, Xiao-gang Peng and Man Yuan — 2019/1/14 0:00:00

Objective To observe the lipid-lowering effect of Typha angustifolia extract (TAE) in Sprague Dawley rats. Methods The rats were randomly divided into six groups, including the hypercholesterolemic diet (HYD) group. Ad libitum food was given to the animals for 60 d to establish dyslipidemia models. The rats were received an ig administration of 1 mL medicine daily. After 120 d the animals were sacrificed by cervical dislocation; Blood was collected to measure total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), nitric oxide (NO), and endothelin (ET) plasma concentration; Livers were collected to measure ApoE mRNA and protein expression; And morphologic changes of aorta ventralis tissue were observed. Results Compared with HYD group, TAE had the ability of reducing TC, TG, LDL-C, NO, and ET (P < 0.01), thereby increasing ApoE mRNA and protein expression of the liver (P < 0.01). Conclusion These results suggest that TAE is capable of effectively decreasing the circulating lipids levels and enhancing the protective effects of artery.

Scutellarin protect human cardiac microvascular endothelial cells by hypoxia-reoxygenation injury involved JAK2/STAT3 signal pathway

2019/1/14 0:00:00

Objective To investigate the antagonistic cell injury effect and molecular mechanism of scutellarin (SCU) in hypoxia reoxygenation (HR) treated human cardiac microvascular endothelial cells (HCMECs). Methods The method of 12 h hypoxia following by 12 h reoxygenation was used to culture HCMECs in vitro to built cell injury model. The groups were divided into control group, model (HR) group, and HR + SCU (0.1 μmol/L, 1 μmol/L, and 10 μmol/L) group. The cell viability was determined by MTT, and oxidative stress was detected by malondialdehyde (MDA) levels by biochemical assay kit. Protein expression of JAK2/p-JAK2 and STAT3/p-STAT3 were evaluated by Western blot. Results The results of MTT and MDA showed that HR decreased the cell viability (P < 0.05) and increased MDA level significantly (P < 0.05), SCU played a contrary role in these processes. Western blot analysis indicates that, the expression of JAK2 and p-JAK2, STAT3, and p-STAT3 were increased in model group when compared with control group (P < 0.05); compared with model group, their expression were reduced by SCU (P < 0.05). Conclusion SCU took a protective effect on HR-treated HCMECs, and the molecular mechanism may be associated with the inhibition of JAK2/STAT3 signal transduction pathway.

Anti-inflammatory activities of gentiopicroside against iNOS and COX-2 targets

2019/1/14 0:00:00

Objective To isolate high-purity gentiopicroside from the Chinese herbal Gentiana officinalis and investigate its anti-inflammatory activity against iNOS and COX-2 targets. Methods The purity and structures of gentiopicroside was determined by HPLC, IR, NMR, and MS. We investigated the anti-inflammatory effects of gentiopicroside by in vivo， in vitro and molecular experiments. Results In vitro experiment results showed that gentiopicroside inhibited nitric oxide (NO), prostaglandin E2 (PGE2), and interleukin-6 (IL-6) production in mouse macrophages RAW 264.7 stimulated by lipopolysaccharide. In vivo experiment found xylene-induced mouse ear swelling was inhibited by gentiopicroside with an inhibition rate of 34.17%. Molecular docking of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) with gentiopicroside showed that hydrogen bonds (H-bonds) were formed between the sugar fragments in gentiopicroside structure with Tyr355, Ser353, Leu352, Ser530, Arg120, and His90 of COX-2, and Glu377, Asp382, Tyr373, Tyr347, Gln263, Asn370, and Gly371 of iNOS. Thus, gentiopicroside had a lower docking score and displayed satisfactory anti-inflammatory activities. Conclusion These results suggested that the mechanism of anti-inflammatory activity of gentiopicroside was associated with the downregulation of inflammatory cytokines, such as NO, PGE2, and IL-6, and the suppression of iNOS and COX-2. Therefore, gentiopicroside is a potential and selective iNOS and COX-2 inhibitor.

A new coumarin glucoside ester from seeds oil leavings of Xanthoceras sorbifolia

2019/1/14 0:00:00

Objective: To study the coumarin from the seeds oil leavings of Xanthoceras Sorbifolia. Methods: The chemical constituents were isolated and purified by combination of silica gel, macroporous resin, Sephadex LH-20, and ODS column chromatography. Their structures were elucidated by spectral and chemical methods. The cytotoxicity of the new compound against ten selected human cancer cell lines was assayed. Results: Five comarins were isolated and identified as fraxetin-7-O-β-D-[6'-(3''-hydroxyl-3''- methylglutaryl)] glucopyranoside (1), fraxoside (2), fraxetin (3), scopoline (4), and esculetin (5), respectively. Conclusion: Compound 1 is a new compound. Unfortunately, this compound exhibited no cytotoxicity with tested cell lines.

Combined application of extended depth of field imaging, image stitching, and polarized microscopy techniques in identification of Spatholobus suberectus

2020/11/9 13:45:56

Objective As traditional techniques for microscopic identification of Chinese medicines currently lack objective and high-quality reference images, here we developed a systemic procedure to be used in microscopic identification of Chinese medicines, which would lead to more objective, effective and accurate identification process. Methods Spatholobi Caulis (Jixueteng in Chinese) was used as the specimen in the development of such procedure. Jixueteng samples were microscopically examined in bright- and dark-field microscopy. Microscopic images were obtained by regular, EDF, and image stitching techniques. Results The microscopic images of the characteristics in pulverized Jixueteng were captured, thanks to EDF imaging and image stitching techniques which allowed the detailed and full sighting of each characteristic to be obtained simultaneously. Different layers in anatomical transverse section, including cork, phelloderm, cortex, phloem, cambium, xylem and pith, were distinctively observed. Moreover, by comparing images of bright- and dark-field microscopy, birefringent and non- birefringent components could readily be distinguished. Conclusion With application of the developed procedure, high-definition, panoramic, microscopic images were acquired, which could be used as the reference images for microscopic identification of Chinese medicines.

Complete chloroplast genome sequence of Amomum villosum and comparative analysis with other Zingiberaceae plants

Li Yang 1, Chong Feng 1, Miao-miao Cai 1, Jie-hu Chen 2, Ping Ding 1 — 2020/11/9 13:45:56

Objective Amomum villosum (AV) is an herb whose dried fruit has been extensively used in modern medicine to treat digestive system diseases such as dysentery, vomiting and abdominal pain. This paper aims to supplement chloroplast (cp) genomic resources and to be used in phylogenetic studies and identification of AV related plants. Methods High-throughput sequencing technology was used to determine the complete sequence of the AV cp genome, and the sequence was then compared with three related species. Results The genome size of AV we obtained was 163,968 bp with an obvious tetrad structure. The AV cp genome was observed to contain 125 unique genes and 81 simple sequence repeat (SSRs) had been determined and the majority of which were adenine-thymine (AT)-rich. Comparative analysis of genome sequence of four ginger plants showed that the atpF, clpP and rpl32 genes are potential markers for identifying Amomum species. Phylogenetic analysis suggested that AV was closely related to A. kravanh and A. compactum. Conclusion These results have brought useful genetic resources for further identification researches, DNA barcoding, resolving taxonomy and understanding the evolutionary mode of Zingiberaceae cp genome.

Enzymatic synthesis of myricetin 3-O-galactoside through a whole-cell biocatalyst

Zhen Xu1,2, Wei-qing He3, Chun-sheng Liu2, Jian-qiang Kong1 — 2020/11/9 13:45:56

Objective: Myricetin 3-O-galactoside is an active compound with pharmaceutical potential. The insufficient supply of this compound becomes a bottleneck in the druggability study of myricetin 3-O-galactoside. Thus, it is necessary to develop a biosynthetic process for myricetin 3-O-galactoside through metabolic engineering. Methods: Two genes OcSUS1 and OcUGE1 encoding sucrose synthase and UDP-glucose 4-epimerase were introduced into BL21(DE3) to reconstruct a UDP-D-galactose (UDP-Gal) biosynthetic pathway in Escherichia coli. The resultant chassis strain was able to produce UDP-Gal. Subsequently, a flavonol 3-O-galactosyltransferase DkFGT gene was transformed into the chassis strain producing UDP-Gal. An artificial pathway for myricetin 3-O-galactoside biosynthesis was thus constructed in E. coli. Results: The obtained engineered strain was demonstrated to be capable of producing myricetin 3-O-galactoside, reaching 29.7 mg/L. Conclusion: Biosynthesis of myricetin 3-O-galactoside through engineered E. coli could be achieved. This result lays the foundation for the large-scale preparation of myricetin 3-O-galactoside.

Total alkaloids of Sophora alopecuroides- and matrine-induced reactive oxygen species impair biofilm formation of Staphylococcus epidermidis and increase bacterial susceptibility to ciprofloxacin

Fang Jiaa, Mei-yang Sunb, Xiang-jun Zhanga, Xue-zhang Zhoua — 2020/11/9 13:45:56

Antihyperglycemic and antihyperlipidemic activities of wild musk melon (Cucumis melo var. agrestis) in streptozotocin-nicotinamide induced diabetic rats

2020/11/9 13:45:56

Objective: Wild musk melon (Cucumis melo var. agrestis, CMA) is one of the edible plants form Tamil Nadu. Traditionally, this plant was used as diabetic diet (leaves of CMA with Momordica charantia leaves), but there is no scientific report on antidiabetic action of this plant material. Hence, the current research work was designed to evaluate its evaluate the anti-hyperglycemic and antihyperlipidemic effect of hydroalcoholic extract of CMA leaves (HALEC) in streptozotocin (STZ)-nicotinamide (NIC)-induced diabetic rats. Methods: Diabetes was induced by administration of STZ (60 mg/kg, i.p.) after 15 min of NIC (120 mg/kg i.p.) administration. The diabetic rats were treated with HALEC (300 and 600 mg/kg, p.o., respectively) for 21 d. Results: After the management with HALEC, blood glucose, HbA1c levels, total cholesterol, LDL cholesterol, triglycerides levels, glycogen phosphorylase and glucose-6-phosphatase levels were significantly diminished in diabetic rats. However, haemoglobin level, HDL cholesterol, liver glycogen, total protein, hexokinase, glucose-6-phosphate dehydrogenase levels were significantly increased in HALEC treated diabetic rats. The histopathological studies of the pancreas in HALEC-treated diabetic rats showed almost normal appearance. L6 cell line study revealed the increased glucose uptake activity of HALEC. High performance thin layer chromatography (HPTLC) analysis confirms the presence of active principles such as rutin, gallic acid and quercetin in HALEC. Conclusion: The results indicated that HALEC possess significant antihyperglycemic and antihyperlipidemic activity in STZ-NIC-induced type II diabetic rats with protective effect. This research work will be useful for the isolation of active principles and development of herbal formulation in phytopharmaceuticals.

In vitro and in silico anti-oxidant, cytotoxicity and biological activities of Ficus benghalensis and Duranta repens

Pukar Khanal, Basanagouda M. Patil — 2020/11/9 13:45:56

Objective: To report in vitro anti-oxidant activity and cytotoxicity of hydroalcoholic extract of Ficus benghalensis (bark) and Duranta repens(whole plant), and present the probable biological spectrum of major anti-oxidants from both plants. Methods: The coarse powder of both plants was first extracted with 70% ethanol (maceration) and followed by 99% ethanol (Soxhlet-extraction). Anti-oxidant activity of the extracts was evaluated by using DPPH, H2O2, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), NO scavenging assay, total antioxidant capacity, cupric reducing antioxidant capacity (CUPRAC), and metal chelating assay. Cytotoxicity of both extracts was evaluated by using MTT assay in both tumor and normal cell lines i.e. Chinese hamster ovary cells (CHO) and A549 cells. Biological activity of individual anti-oxidant compounds was identified by using prediction of activity spectra for substances and a docking study was performed by using autodock4.0. Results: Hydroalcoholic extract of F. benghalensis and D. repens showed the highest free radical scavenging capacity (ABTS) and chelating capacity respectively. Both extracts showed minimum cytotoxicity in normal cell lines compared to tumor cell lines. Computer imitation hits reflected the multiple biological activities agreeing with the folk use and some scientific reports. Further, we found the potent binding affinity of predicted anti-oxidant compounds with multiple protein molecules involved in oxidative stress. Conclusion: The present study reports the probable anti-oxidant mechanism for two folk agents and also presents probable pharmacological activities via computer simulations.

Long-term consumption of virgin coconut (Cocos nucifera) oil diet impairs learning and memory in CD1 mice

2020/11/9 13:45:56

Objective: Virgin coconut oil (VCO) has been used in the management of dementia in Alzheimer’s disease (AD). Therefore, this research investigated the effect of long-term consumption of VCO diet on learning and memory in CD1 mice. Methods: Thirty male CD1 mice (divided into three groups, n = 10) were fed with standard rodent chow (control), 5% and 20% VCO diets (respectively) for 28 d. The Morris Water Maze (MWM) test was used to test the effect of VCO on visuo-spatial learning and memory, while the Novel Object Recognition Test (NORT) was used to measure short- and long-term recognition memory. Results: Learning performance of mice did not differ in the MWM. During the probe trial, duration in the retention quadrant and annulus crossings were lower (P < 0.05) in the 5% and 20% VCO diet groups compared to the control diet group, showing that VCO impaired visuo-spatial memory. During the NORT, mice showed more total approaches in the 20% VCO diet group (P < 0.05) compared to control and the 5% VCO diet groups during the short-term memory test. During the long-term memory retention test, the total approaches were also higher in the 20% VCO group compared to control and 5% VCO group (P > 0.05). The discrimination index was also lower in the 20% VCO group compared to control and 5% VCO diet groups indicating impaired long-term cognitive memory in mice given 20% VCO diet. Histological examination of brains showed damage within the CA1 pyramidal cell layer of the hippocampus in the 20% VCO diet group, in line with the behavioural observations. Conclusion: Long-term consumption of virgin coconut oil diet impairs memory in mice.

Shenqi Xingnao Granules ameliorates cognitive impairments and Alzheimer’s disease-like pathologies in APP/PS1 mouse model

2020/11/9 13:45:56

Objective: Alzheimer's disease?(AD) is along with cognitive decline due to amyloid-β (Aβ) plaques, tau hyperphosphorylation, and neuron loss.?Shenqi Xingnao Granules (SQXN), a traditional Chinese medicine, significantly ameliorated the cognitive function and daily living abilities of patients with AD. However, till date, no study has investigated the mechanism of action of SQXN on AD. The present study aimed to verify the effects of SQXN treatment on cognitive impairments and AD-like pathologies in APP/PS1 mice. Methods: Four-month-old?APP/PS1?transgenic (Tg) mice were randomly divided into a model group and SQXN-treated (3.5, 7, 14 g/kg per day) groups. Learning-memory abilities were determined by Morris water maze and object recognition test. All mice were sacrificed and the brain samples were collected after 75 d. The soluble Aβ contents were detected by Elisa kit; The levels of expression of NeuN, APP, phosphorylated tau and related protein were measured by Western blotting; The inflammation factors were detected by the proinflammatory panel kit. Results: Four-month-old?APP/PS1?mice were administered SQXN by oral gavage for 2.5 months. Using the Morris water maze tests and Novel object recognition, we found that SQXN restored behavioral deficits in the experimental group of Tg mice when compared with the controls. SQXN also inhibited neuronal loss?(NeuN?marker). SQXN treatment decreased soluble Aβ42 through inhibiting the expression of sAPPβ and BACE-1 without regulating full-length amyloid precursor protein (FL APP). Insulin degrading enzyme (IDE), the Aβ degrading enzyme, were increased by SQXN. In addition, SQXN reduced hyperphosphorylated tau protein levels and prevented excessive activation of p-GSK-3β in the brain of APP/PS1 mice. Compared with APP/PS1 transgenic negative mice, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-12p70, KC/GRO and TNF-α were not obviously changed in the brain of 6.5-month-old APP/PS1?transgenic (Tg) mice. However, SQXN could inhibited the expression of IL-2. Conclusion: These results demonstrate that SQXN ameliorates the cognitive impairments in APP/PS1 mice. The possible mechanisms involve its inhibition of neuronal loss, soluble Aβ deposition, tau hyperphosphorylation and inflammation.

Quantitative determination of multi-class bioactive constituents for quality assessment of ten Anoectochilus, four Goodyera and one Ludisia species in China

2020/11/9 13:45:56

Objective: To establish multi-class bioactive constituents’ determination of ten Anoectochilus, four Goodyera and one Ludisia species, and provide reference for the improvement of their quality control. Methods: HPLC-ELSD and phenol-sulphuric acid methods were used for the quantitative determination of lactone glycosides (kinsenoside and its diastereoisomer, gooderoside A) and polysaccharides, respectively, while an efficient iHPLC–MS/MS method was established for rapid determination of other minor constituents in ten Anoectochilus species and five related species. Results: The contents of kinsenoside, goodyeroside A, polysaccharides and flavonoids varied notably almost in all tested samples, including both wild plants and tissue cultures. In particular, kinsenoside was the major lactone glycoside in A. roxburghii, A. formosanus, A. xingrenensis, A. nandanensis, A. brevilabris and A. burmannicus, whereas goodyeroside A was the predominant constituent in A. lylei, A. longilobus, A. elatus, A. zhejiangensis, G. schlechtendaliana, G. biflora, G. yangmeishanensi, G. repens and Ludisia discolor. Conclusion: Our present study suggested that A. lylei, A. longilobus, A. elatus, A. zhejiangensis, Ludisia discolor and Goodyera species cannot be used as alternatives for A. roxburghii, and goodyeroside A may be reasonably used as a diagnostic marker for distinguishing A. roxburghii from A. lylei, A. longilobus, A. elatus and A. zhejiangensis, Goodyera and Ludisia species. The established method thus could be potentially used for the quality evaluation and control of Anoectochilus and some related species.

UGT88B2: A promiscuous O-glycosyltransferase from Carthamus tinctorius

Song-yang Suia, Rui-mingqian Guoa, Ke-bo Xieb, Lin Yanga — 2020/11/9 13:45:57

Objective: In order to obtain new glycosyltransferases with highly efficient catalysis, the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined. Methods: A new glycosyltransferase gene (UGT88B2) with full length was obtained by PCR and further transformed into Escherichia coli for heterologous expression. The catalytic activity of recombinant UGT88B2 was determined by HPLC-MSn. The structures of representative catalytic products were elucidated by MS and NMR. Results: UGT88B2 exhibited catalytic promiscuity and various patterns in glycosylation of flavonoids with high efficiency. Conclusion: A new glycosyltransferase named UGT88B2 was successfully mined and can be employed as enzymatic tools in glycosylation of flavonoids.

Comparative pharmacokinetics of baicalin and geniposide in juvenile and adult rats after oral administration of Qingkailing Granules

2020/11/9 13:45:57

Objective: To explore the effect of age on Qingkailing Granules disposition by comparing the pharmacokinetics of geniposide and baicalin in juvenile and adult rats. Methods: A simple and rapid LC-MS/MS method was developed and validated to simultaneously determine geniposide and baicalin in rat plasma after a simple protein precipitation. The analytes were separated on an Agilent ZORBAX Extend-C18 column. The mobile phase consisted of acetonitrile and water with 0.1% (volume percent) formic acid at a flow rate of 0.6 mL/min. The ionization was conducted using an ESI source in negative ion mode. Multiple reaction monitoring was used for quantification at transitions of m/z 445.0 → m/z 268.9 for baicalin, m/z 433.2 → m/z 225.0 for geniposide, m/z 431.0 → m/z 341.0 for vitexin (IS). Juvenile and adult rats were administrated Qingkailing Granules (3 g/kg) orally. Plasma concentrations of baicalin and geniposide were determined by LC-MS/MS. Results: The linear ranges of the analytes were 1–1000 ng/mL for baicalin and 2–2000 ng/mL for geniposide. The method was successfully applied to compare the pharmacokinetics of the analytes between juvenile and adult rats after oral administration of Qingkailing Granules. AUC was bigger in adult rats, while t1/2 was longer in juvenile rats. Conclusion: These results suggested that the absorption and elimination of baicalin and geniposide in juvenile rats was lower than that in adult rats. Additional attention should be paid to the pharmacokinetic difference when Qingkailing Granules were used in children.

Yanangdaengin, a dihydrochalcone glucoside galloyl ester as active antioxidative agent from leaves of Lysiphyllum strychnifolium (syn. Bauhinia strychnifolia)

2020/11/9 13:45:57

Objective: To isolate and identify the major bioactive components from the leaves of Lysiphyllum strychnifolium, an indigenous herb used in traditional Thai medicine for detoxification, longevity, and some other health related issues. Methods: Comparative HPLC analyses of the crude extracts from three provenances were carried out for an overview of characteristic compound profiles. Isolation of the major compounds was undertaken with chromatographic methods. Chemical structures were elucidated by NMR spectroscopic techniques and mass spectrometry. DPPH scavenging assay was carried out to determine the free radical scavenging activity of isolated compounds. Results: Yanangdaengin (3), a dihydrochalcone glucoside galloyl ester, has been isolated together with its corresponding dihydrochalcone glucoside trilobatin (2) as major compounds from the leaves of L. strychnifolium. Additionally, gallic acid (1) was co-chromatographically identified. Free radical scavenging activity of isolated compounds were determined. Compound 3 exhibited higher free radical scavenging activities in comparison to Trolox and quercetin. Conclusion: The isolated compounds could be used as chemical markers for quality assessment. The present work could promote the quality control and herbal medicinal product development of this plant.

Cymbogogoncitratus aqueous leaf extract attenuates neurobehavioral and biochemical changes induced by social defeat stress in mice

2020/7/19 22:28:19

Objective Psychosocial stress has been implicated in the genesis of psychiatric disorders such as memory deficits, depression, anxiety and addiction. Aqueous leaf extract of Cymbopogon citratus (CYC) otherwise known as lemongrass tea has antidepressant, anxiolytic and anti-amnesic effects in rodents. This study was designed to evaluate if C. citratus could reverse the neurobehavioral and biochemical derangements induced by social defeat stress (SDS) in the resident/intruder paradigm. Methods Intruder male mice were divided into five groups (n = 7): group 1 received saline (10 mL/kg, p.o.; non-stress control), group 2 also received saline (10 mL/kg, p.o.; SDS control) while groups 3?5 had C. citratus (50, 100 and 200 mg/kg, p.o.) daily for 14 d. The SDS was carried out 30 min after each treatment from day 7 to day 14 by exposing each intruder mouse in groups 25 to a 10 min confrontation in the home cage of an aggressive resident counterpart. The neurobehavioral features (spontaneous motor activity-SMA, anxiety, memory, social avoidance and depression were then evaluated. The concentrations of nitrite, malondialdehyde and glutathione as well as acetylcholinesterase activity in the brain tissues were also determined. Results C. citratus (50, 100 and 200 mg/kg) attenuated hypolocomotion, heightened anxiety, depressive-like symptom, memory deficit and social avoidance induced by SDS. The altered levels of oxidative stress and acetyl-cholinesterase in SDS-mice were positively modulated by C. citratus. Conclusion The results of this study suggest that C. citratus might mitigate psychosocial stress-induced neurologic diseases in susceptible individuals.

Sterols possess immunomodulatory property: Activity guided isolation and in vitro screening of phytoconstituents of Pongamia glabra and Ficus glomerata

2020/7/19 22:28:19

Objective: To isolate the phytoconstituents from the methanolic extracts of the stem bark of Pongamia glabra and Ficus glomerata., characterize spectroscopically and screen for in vitro immunomodulatory activity on human neurophils. Methods: A flavonoid (PGF) and an alkaloidal compound (PGA) from the extract of P. glabra and a steroidal compound (FGS) and tannin fraction (FGT) from the extract of F. glomerata were isolated using column chromatography technique and were subjected for the spectroscopic (FT-IR, 1HNMR and LC-MS) and TLC studies to identify the compounds. The isolated compounds were screened for in vitro immunomodulatory activity on human neutrophils using nitroblue tetrazolium (NBT) dye test, phagocytosis of Candida albicans and neutrophil locomotion and chemotaxis assay at the concentration range of 100, 50, 25, 12.50 and 5.00 μg/mL. Results: From the spectroscopic and TLC studies data, the isolated compounds were identified as glabrin (PGA), karanjin (PGF), β-sitosterol (FGS), and tannin fraction (FGT). The isolated compounds PGA, PGF, FGS, and FGT exhibited significant (P < 0.05) in vitro immunomodulatory activity in all the parameters studied. Conclusion: The steroidal compound, i.e. FGS was found to be more immunopotent than all constituents alkaloid, flavonoid and tannins. Hence, these constituents could be attributed to the immunomodulatory property of the plants. Objective: To isolate the phytoconstituents from the methanolic extracts of the stem bark of Pongamia glabra and Ficus glomerata., characterize spectroscopically and screen for in vitro immunomodulatory activity on human neurophils. Methods: A flavonoid (PGF) and an alkaloidal compound (PGA) from the extract of P. glabra and a steroidal compound (FGS) and tannin fraction (FGT) from the extract of F. glomerata were isolated using column chromatography technique and were subjected for the spectroscopic (FT-IR, 1HNMR and LC-MS) and TLC studies to identify the compounds. The isolated compounds were screened for in vitro immunomodulatory activity on human neutrophils using nitroblue tetrazolium (NBT) dye test, phagocytosis of Candida albicans and neutrophil locomotion and chemotaxis assay at the concentration range of 100, 50, 25, 12.50 and 5.00 μg/mL. Results: From the spectroscopic and TLC studies data, the isolated compounds were identified as glabrin (PGA), karanjin (PGF), β-sitosterol (FGS), and tannin fraction (FGT). The isolated compounds PGA, PGF, FGS, and FGT exhibited significant (P < 0.05) in vitro immunomodulatory activity in all the parameters studied. Conclusion: The steroidal compound, i.e. FGS was found to be more immunopotent than all constituents alkaloid, flavonoid and tannins. Hence, these constituents could be attributed to the immunomodulatory property of the plants. Objective: To isolate the phytoconstituents from the methanolic extracts of the stem bark of Pongamia glabra and Ficus glomerata., characterize spectroscopically and screen for in vitro immunomodulatory activity on human neurophils. Methods: A flavonoid (PGF) and an alkaloidal compound (PGA) from the extract of P. glabra and a steroidal compound (FGS) and tannin fraction (FGT) from the extract of F. glomerata were isolated using column chromatography technique and were subjected for the spectroscopic (FT-IR, 1HNMR and LC-MS) and TLC studies to identify the compounds. The isolated compounds were screened for in vitro immunomodulatory activity on human neutrophils using nitroblue tetrazolium (NBT) dye test, phagocytosis of Candida albicans and neutrophil locomotion and chemotaxis assay at the concentration range of 100, 50, 25, 12.50 and 5.00 μg/mL. Results: From the spectroscopic and TLC studies data, the isolated compounds were identified as glabrin (PGA), karanjin (PGF), β-sitosterol (FGS), and tannin fraction (FGT). The isolated compounds PGA, PGF, FGS, and FGT exhibited significant (P < 0.05) in vitro immunomodulatory activity in all the parameters studied. Conclusion: The steroidal compound, i.e. FGS was found to be more immunopotent than all constituents alkaloid, flavonoid and tannins. Hence, these constituents could be attributed to the immunomodulatory property of the plants.

Substance basis of warm nature of Poria cocos

Lin-lin Liu, Ya-nan Wang, Cheng Tan, Xue-ying Han, De-qiang Dou — 2020/7/19 22:28:19

Objective The research indicated that the nature of Chinese medicine is mainly related to body’s substance and energy metabolism. The purpose of the study is to elucidate the substance basis for warm nature of Poria cocos (called Fuling (FL) in Chinese). Methods In terms of the effects of its separated fractions on the substance and energy metabolism in rat models of cold-deficiency with Aconiti Lateralis Radix Praeparata (called Fuzi (FZ) in Chinese), with hot nature, as reference drug. Biochemical indexes in the material metabolism, energy metabolism, endocrine system, nervous system and nucleotide system were determined, then analyzed by additive, cluster and principal component analysis (PCA). Results The medicinal natures of oligosaccharides and amino acids fractions were attributable to plain and crude polysaccharides, volatile oils and triterpenoids fractions were attributable to mild warm. Conclusion The nature of FL was regarded as mild warm based on the old records of Chinese medicine and fractions of crude polysaccharides, volatile oils and triterpenoids might be the main substance basis for the warm nature of FL. It is the first time that substance basis of FL was elucidated from view point of medicinal nature.

Phytochemicals and antioxidant activity of alcoholic/hydroalcoholic extract of Trifolium pratense

Mohsen Akbaribazm, Mohammad Rasoul Khazaei, Mozafar Khazaei — 2020/7/19 22:28:19

Objective: Trifolium pratense has many healing properties, including fewer complications of menopause, cancer cell suppression, reducing blood glucose and lipids, as well as cardiovascular beneficial effects. The purpose of this study was to identify the phytochemical and mineral composition of T. pratense. Methods: Plant aerial parts were harvested and dried, and then hydroalcoholic and alcoholic extracts were prepared. Gas chromatography–mass spectrometry (GC-MS) analytical method was used to identify volatile compounds then liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) was used to identify polyphenols and the mineral elements were identify by inductively coupled plasma atomic emission spectrometer/ICP-AES and scanning electron microscope-energy-dispersive X-ray spectroscopy (SEM-EDS) methods. Total phenolic content (TPC) was determined based on colorimetric method, and total flavonoid content (TFC) was established based on the folin-chiocalteau reagent. Furthermore, two assays (DPPH and FRAP) were used to measure the antioxidant capacity of T. pratense ethanolic extract. Results: A total of 37 polyphenol and 107 peaks were identified by LC-ESI-MS analysis, and the GC/MS method also detected 21 volatile compounds, the most important of which were methylcyclopentane, dimethylpentanal and hexadecanol. A total of 18 mineral elements, including K, Mg, Al, Si, Zn, Ni, Cu, Se, Co, Fe, Mn, and Ca in the plant, were identified ICP-AES and SEM-EDS analysis. Conclusion: T. pratense has many therapeutic compounds such as polyphenol (isoflavone and flavonoids), volatile compounds, and essential mineral elements, which can be formulated purely and used in the pharmaceutical and traditional medicine industries.

Antioxidation and active constituents analysis of flower residue of Rosa damascen

2020/7/19 22:28:19

Objective: To make full usage of resource and turn waste into treasure, the chemical constituents and bioactivity were firstly investigated on Damask rose (Rosa damascena) flower residue (DRFR). Methods: DPPH and ABTS experiments were applied to assess the antioxidant activity of DRFR. Then, column chromatography was used to purify compounds from an antioxidation extract (DRFR-A), and the chemical structure was identified using NMR. The total phenolic acid content was measured by Folin-Ciocalteu colorimetric method, and the content of gallic acid of the indicator ingredient was detected by HPLC. Results: DRFR-A was found to show a high activity both on DPPH (IC50: 2.760 μg/mL) and ABTS (IC50: 2.258 μg/mL) compared to positive control VC. Ten compounds were isolated and identified as quercetin (1), kaempferol (2), gallic acid (3), protocatechuic acid (4), pyrogallic acid (5), 2-phenylethyl 3,4,5-trihydroxybenzoate (6), methyl gallate (7), p-hydroxybenzoic acid (8), p-hydroxyphenethyl alcohol (9) and astragalin (10) from DRFR-A. Among them, pyrogallic acid, 2-phenylethyl-3, 4, 5-trihydroxybenzoate, p-hydroxybenzoic acid and p-hydroxyphenethyl alcohol are obtained from the plant for the first time. The content of total phenolic acids and gallic acid, main ingredient in DRFR-A was determined as 63.73% and 24.67%, respectively. Conclusion: This study provides a reliable data and lays the foundation for the development and utilization of rose residue, and hence for the full utilization of rose resources.

New phenolic constituents obtained from Polygonum multiflorum

2020/7/19 22:28:19

Objective: To isolate the phenolic compounds obtained from the dried roots of Polygonum multiflorum and investigate their pharmacological activities. Methods: The chemical constituents were isolated and purified by combining them with a macroporous resin (DM-8), MCI gel, and Sephadex LH-20 and by performing ODS column chromatography. Their structures were elucidated by 1D and 2D NMR analyses, as well as mass spectrometry. The isolated compounds were evaluated to determine their hepatoprotective and α-glucosidase inhibitory activities in vitro. Results: Two phenolic compounds, namely, polygonimitin E (1) and polygonimitin F (2), were isolated from the dried roots of P. multiflorum. Compound 2 (10 μmol/L) only showed moderate hepatoprotective activity against N-acetyl-p-aminophenol (APAP)-induced HepG2 cell damage. Unfortunately, these two compounds exhibited no α-glucosidase inhibitory activity. Conclusion: Compounds 1 and 2 were new compounds. Compound 2 could be one of the potential hepatoprotective constituents of P. multiflorum.

Critical process parameter identiﬁcation of manufacturing processes of Astragali Radix extract with a weighted determination coeﬃcient method

2020/5/18 17:53:42

Objective: Critical process parameters (CPPs) identification is an important step of the implementation of quality by design (QbD) concept. There are many CPP identification methods, such as risk analysis method, sensitivity analysis method, multiple linear regression method, standard partial regression coefficient (SPRC) method, and so on. The SPRC method can consider multiple process critical quality attributes (CQAs) simultaneously, but the determination of CPP number is subjective. Therefore, new CPP identification method is still required. Methods: The manufacturing process of Astragali Radix extract, which contained water reflux extraction, concentration, and ethanol precipitation, was used as an example. First, the multiple process CQAs were determined to be the yield of pigment, dry matter, sugars, and active ingredients. Second, the potential CPPs were determined by a knowledge organization method. Plackett-Burman designed experiments were then performed. A weighted determination coefficient () method was presented to identify CPPs. In this method, the importance of different CQAs was considered. Process parameters were removed one-by-one according to their importance index. The decrease in was used to characterize the importance of the removed parameter. If the decrease of was less than a preset threshold, the removed parameter was not a CPP. Results: During the manufacturing process of Astragali Radix extract, the potential CPPs determined by the knowledge organization method were water consumption, reflux extraction time, extraction frequency, ethanol content, ethanol consumption, and concentration endpoint. Reflux extraction time, the first ethanol consumption, the second ethanol consumption, and the second ethanol precipitation refrigeration temperature were found to be CPPs using the weighted determination coefficient method with the threshold of 10%. Conclusion: Using the weighted determination coefficient method, CPPs can be determined with all the CQAs considered based on their importance. The determination of CPP number is more objective compared with the SPRC method.

Mining lycodine-type alkaloid biosynthetic genes and genetic markers from transcriptome of Lycopodiastrum casuarinoides

Gang Chena,b, Qiu-mei Lina, Lin Zenga, Yi-ping Zoua — 2020/5/18 17:53:42

Alhagi maurorum aqueous extract protects against norfloxacin-induced hepato-nephrotoxicity in rats

Hesham A. Khalifa1, Shimaa I. Shalaby2, Ahmed S. Abdelaziz1 — 2020/5/18 17:53:42

Objectives: While the protective effects of Alhagi maurorum have been shown against various ailments, its role against norfloxacin-induced adverse effects has not been studied. The current study was conducted to determine the effect of A. maurorum aqueous extract against norfloxacin-induced side effects in rats. Methods: Twenty four male albino rats were randomly assigned into four groups, which received normal saline, norfloxacin (50 mg/kg b.wt orally once a day), A. maurorum aqueous extract (300 mg/kg b.wt orally once a day), and norfloxacin with A. maurorum aqueous extract by the same previous mentioned dosages. Blood samples were collected for hematological examination to evaluate liver and kidney function tests. Hepatic and renal tissue samples were obtained to assess antioxidant activity and histopathological examination. Results: A. maurorum aqueous extract significantly ameliorated norfloxacin-induced elevation in tissue malondialdehyde, and reduction in tissue antioxidant enzymes such as catalase, glutathione peroxidase, and superoxide dismutase activities as well as reduced glutathione concentration. Concurrent administration of A. maurorum aqueous extract with norfloxacin significantly reduced serum alkaline phosphatase, aminotransferases, urea, creatinine, and uric acid and increased RBCs count, Hb concentration, PCV, leucocyte, and monocyte counts compared with the norfloxacin-treated group. Co-administration of A. maurorum aqueous extract with norfloxacin prevented the degenerative changes induced by norfloxacin alone in liver and kidney tissues. The phytochemical profile of the extract showed the presence of carbohydrates, alkaloids, saponins, tannins, phenolics, and flavonoids. Conclusion: These findings indicated that A. maurorum possesses potent antioxidant activities and could be used to attenuate norfloxacin-induced side effects.

Methanol extract of Basella alba leaves alleviate stress in rats

2020/5/18 17:53:42

Objective: Stress is becoming an unavoidable threat in recent times, there has been increasing interest by researchers in the use of naturally occurring biologically active compounds with medicinal value to cure body ailments. The present work was carried out to investigate the effect of methanol extract of Basella alba leaves on stress in Wistar rats (Rattus norvegicus). Methods: A total of 35 male rats were used in this study. They were grouped into seven groups of 5 rats each. Group 1 (normal control) was received 10 mL/kg normal saline. Group 2 contained restraint stress rats only. Group 3 contained forced swim stress rats only. Group 4 and 5 were treated with 60 mg/kg of B. alba extract (BAE) thereafter subjected to restraint and forced swim stresses respectively. Group 6 and 7 were treated with 120 mg/kg of BAE thereafter subjected to restraint and forced swim stresses respectively. Stress procedures were carried out at the end of first and third weeks. Results: In the stressed rats, there were significant increases (P < 0.05) in fasting blood glucose and white blood cell count while there were significant decreases in superoxide dismutase activity and glutathione concentration when compared to group 1. There were significant decreases (P < 0.05) in blood glucose and white blood cell count and significant increases in superoxide dismutase and glutathione concentrations in BAE treated rats when compared to group 2 and 3. Some of the significant differences were either dose or duration dependent. Conclusion: In conclusion, results from this research suggest that BAE alleviates hyperglycaemia, chronic activation of immune system and generation of free radicals due to stress in Wistar rats.

Effect of Vigna radiata, Tamarix ramosissima and Carthamus lanatus extracts on Leishmania major and Leishmania tropica: An in vitro study

2020/5/18 17:53:42

Objective: Current therapy strategies of leishmaniasis have some problems such as high cost, toxicity and side effects. Plant extracts can be a source of drugs to control leishmaniasis. In this study, the effect of hydroalcoholic and chloroformic extracts of Vigna radiata, Tamarix ramosissima, and Carthamus lanatus on Leishmania major and L. tropica was studied. Methods: The plant samples were collected from west of Iran and their extracts were prepared. Anti-promastigote activity assay of all extracts was done using tetrazolium-dye assay. Results: Only high concentrations of V. radiata and C. lanatus were able to inhibit Leishmania, while both high and low concentrations of T. ramosissima had antileishmanial effect. No difference was observed between hydroalcoholic with chloroformic extract of each plant. Conclusion: Altogether, the results revealed the antileishmanial activity of T. ramosissima extracts against L. major and L. tropica, indicating its potential as an antileishmanial agent.

Human pulp tissue dissolution ability of different extracts of Sapindus mukorossi: An in vitro study

?znur Gü?lüer a, Esin Akarsu b, Emre Yavuz b, Kür?at Er a, Alper Ku?tarc? a — 2020/5/18 17:53:42

Objective: Due to the many negative properties of sodium hypochlorite used in current root canal treatment, interest in biocompatible natural agents is increasing day by day. The aim of this study was to evaluate whether various extract solutions of Sapindus mukorossi have dissolution effects on human pulp tissues. Methods: Primarily powder extracts were obtained by extracting fruit shells of S. mukorossi in different solvents (ethanol, methanol, buthanol and distilled water). The test solutions were prepared and randomly separated into six groups with 10 samples in each group: ethanol extract, methanol extract, butanol extract, distilled water extract of S. mukorossi, sodium hypochlorite (NaOCl) and the control group. Among these, S. mukorossi extracts were separated into two subgroups, depending on their concentration level (50 μg/mL and 100 μg/mL). The pulp tissues of freshly extracted human molars were used for dissolution test. The weights of the pulpal tissues were measured and recorded for two times after the samples were placed in the solutions. Statistical analysis for all descriptive statistics was performed using SPSS 22 (P < 0.05). Results: Our results showed that maximum percent yield of preparation was obtained in methanol extract of S. mukorossi. Among all of the groups, the best dissolution capacity was seen in the NaOCl group (positive control group). Among S. mukorossi groups, the best tissue solvent solution was found in SMM group at 50 μg/mL and SMB group at 100 μg/mL. Conclusion: The different extracts of S. mukorossi had a capacity to dissolve pulp tissue but this capacity was less than NaOCl. Therefore, further studies will enable the creation of a commercial solution for clinical use by increasing the effectiveness of S. mukorossi while combining it with other endodontic irrigation solutions.

Two new terpenoids from Reduning Injection

2020/5/18 17:53:42

Objective To study the antipyretic and anti-inflammatory constituents from the active fraction of Reduning (RDN) Injection. Methods In this study, the active fraction of RDN injection was screened by the LPS-induced mouse endotoxin shock model. The chemical constituents were isolated by chromatography on HP-20 macroporous adsorptive resins, silica gel, ODS columns and reverse phase MPLC and HPLC repeatedly, and their structures were elucidated based on spectroscopic analysis (HR-ESI-MS, NMR, ECD) and chemical methods. Meanwhile, we evaluated the anti-inflammatory activities of the isolates by measuring their inhibitory effects on TNF-α production in LPS stimulated RAW 264.7 macrophages. Results The 95% ethanol eluate of RDN injection by the macroporous adsorption resin column was proved to be the antipyretic and anti-inflammatory active fraction of this injection. A novel iridoid, named jasminoide A (1), and a new guaiane sesquiterpenoid, named (1R,7R,8S,10R)-7,8,11-trihydroxy-4-guaien-3-one (2), were isolated from Reduning injection, and compound 2 can inhibit TNF-α production with IC50 values of 72.24 μmol/L. Conclusion In this study, Two new terpenoids were isolated from Reduning injection, and compound 2 showed inhibitory activity against LPS-activated TNF-α production in RAW 264.7 cells in vitro.

A new strategy based on acid-alkali complexation for rapidly and accurately fishing phytochemicals in Sennae Folium

2020/5/18 17:53:43

Objective: There are some anthracenone, anthraquinones and flavonones in Sennae Folium which exhibited significant acidity, such as sennoside A/B and sennoside C/D. The current strategy used in separating these components are mainly based on conventional column chromatography which is time consuming, laborious and costly. This study is aimed at exploring a method of precipitation extraction of acid components in Sennae Folium. Using alkaloid as a “hook”, it is reasonable to use the principle of “acid-alkali complexation” to "fish" the acidic components in Sennae Folium. Methods: Isothermal titration calorimeter (ITC) was used to measure the extraction efficiency of different alkaloids. Then, alkaloid determined by ITC was mixed with extracting solution of Sennae Folium to form complex. High performance liquid chromatography coupled with mass spectrometry (HPLC-MS2) was used to investigate the ingredients “fished” by berberine (Ber). The mechanism of “fishing” process was explained by ITC, optical activity, fluorescence spectrometry and scanning electron microscope. Results: The ITC results proved that the choice of “hook” was particularly important in the process of “fishing”. Among the hooks, the fishing efficiency of the isoquinoline alkaloids (Ber) was the highest, reaching 10.3%. Nine ingredients were detected and determined by HPLC-MS2, and the main components were sennoside A/B and sennoside C/D. Based on ITC test of Ber and sennoside A, the combination mechanism of the two ingredients was a chemical reaction with a nearly binding ratio (2:1). Fluorescence and optical properties of the active ingredients were changed after complexation. By scanning electron microscope, we found that two types of components had obviously self-assembled behavior during the formation process. Conclusion: Ber successfully “fished” the main acidic components, sennoside A/B and sennoside C/D, from Sennae Folium. Combined with different characterizations, the “fishing” process was determined as a chemical association reaction induced by electrostatic interaction or π-π stacking. Therefore, with special identification ability, the “fishing” process had the potential of practical application.

Chemical constituents from acid hydrolyzates of Panax quinquefolius total saponins and their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B

2020/5/18 17:53:43

Objective To investigate the hypoglycemic components from the acid hydrolyzates of Panax quinquefolius total saponins，and screen the active compounds by in vitro inhibitory activities to α-glycosidase enzymes and protein tyrosine phosphatase-1B (PTP1B). Methods The hydrolyzates were chromatographed repeatedly over silica gel column, and the structures of the compounds were determined by means of NMR. The in vitro bioassay was performed through the inhibitory effects on α-glucosidase or/and PTP1B. Results Eight compounds were isolated, which identified as 20(S)-panaxadiol (1), (20S,24R)-dammarane-20,24-epoxy-3β,6α,12β,25-tetraol (2), 20(R)-dammarane-3β,12β,20,25-tetraol (3), 20(S)-dammarane-3β,6α,12β,20,25-pentol (4), 20(R)-dammarane-3β,12β,20,25-tetrahydroxy-3β-O-β-D-glucopyranoside (5), β-sitosterol (6), oleanolic acid (7) and 20(S)-protopanaxadiol (8). Compound 5 was ginseng triterpenoid isolated from the acid hydrolysates of total saponins from P. quinquefolius for the first time. In this paper, the possible in vitro inhibitory activities were investigated. Compound 5 exhibited significantly inhibitory activity against α-glucosidase, and the IC50 value [(0.22 ± 0.21) μmol/L] was about 43-fold lower than positive control. For the PTP1B inhibition assay, compound 5 indicated the strongest inhibitory effect with IC50 of (5.91 ± 0.38) μmol/L, followed by compound 4 with IC50 of (6.21 ± 0.21) μmol/L, which were all showed competitive inhibitory pattern by using a Lineweaver-Burk plot. Conclusion These results supported the potential application of dammaranes from acid hydrolyzates of P. quinquefolius total saponins can be used as ingredients of ancillary anti-diabetic agent or functional factor.

A validated HPTLC method for quantitative analysis of morin in Maclura cochinchinensis heartwood

Savita Chewchindaa, Sumet Kongkiatpaiboonb — 2020/5/18 17:53:43

Objective: The study was conducted to develop and validate a high performance thin-layer chromatography (HPTLC)-densitometric method for the quantitative analysis of morin in Maclura cochinchinensis heartwood collected from different locations in Thailand. Methods: HPTLC analysis was performed on an aluminium sheet of silica gel 60 F254 using toluene: ethyl acetate: formic acid (36:12:7, volume percent) as a mobile phase. The densitometric scanning was performed at the wavelength 410 nm. HPTLC method was validated according to ICH guideline. Results: The proposed HPTLC method showed acceptable validation parameters. The content of morin in M. cochinchinensis heartwood collected from eight different provinces in Thailand were in the ranges of 1.53%?2.73%. Conclusion: The simple and sensitive HPTLC method was successfully developed and validated for determination of morin in M. cochinchinensis heartwood. The proposed HPTLC method was found to be simple, fast and inexpensive, and can be used for the routine quality control of raw materials.

Integrated chemical and transcriptomic analyses unveils synthetic characteristics of different medicinal root parts of Angelica sinensis

2020/2/18 16:22:10

Objective: Why are different medicinal parts including heads, bodies and tails of Angelicae Sinensis Radix (ASR) distinct in pharmaceutical activities? Here we explored their discrepancy in chemical constituents and transcriptome. Methods: ASR were separated into three medicinal parts: heads (rootstocks with petiole traces of ASR), bodies (taproots of ASR) and tails (lateral roots of ASR), and chemical and transcriptomic analyses were conducted simultaneously. Results: High performance liquid chromatography (HPLC) fingerprint results showed that five widely used active ingredients (ferulic acid, senkyunolide H, senkyunolide A, n-butylphathlide, and ligustilide) were distributed unevenly in the three ASR medicinal parts. Partial least squares-discriminant analysis (PLS-DA) demonstrated that the heads can be differentiated from the two other root parts due to different amounts of the main components. However, the content of ferulic acid (a main quality marker) was significantly higher in tails than in the heads and bodies. The transcriptome analysis found that 25,062, 10,148 and 29,504 unigenes were specifically expressed in the heads, bodies and tails, respectively. WGCNA analysis identified 17 co-expression modules, which were constructed from the 19,198 genes in the nine sam- ples of ASR. Additionally, we identified 28 unigenes involved in two phenylpropanoid biosynthesis (PB) pathways about ferulic acid metabolism pathways, of which 17 unigenes (60.7%) in the PB pathway were highly expressed in the tails. The expression levels of PAL, C3H, and CQT transcripts were significantly higher in the tails than in other root parts. RT-qPCR analysis confirmed that PAL, C3H, and CQT genes were predominantly expressed in the tail parts, especially PAL, whose expression was more than doubled as compared with that in other root parts. Conclusion: Chemical and transcriptomic analyses revealed the distribution contents and pivotal tran- scripts of the ferulic acid biosynthesis-related pathways. The spatial gene expression pattern partially explained the discrepancy of integral medicinal activities of three medicinal root parts.

Critical pharmaceutical process identification considering chemical composition, biological activity, and batch-to-batch consistency: A case study of notoginseng total saponins

2020/2/18 16:22:10

Objective: Critical pharmaceutical process identification (CPPI) is an important step in the implementa- tion of quality by design concept to traditional Chinese medicines (TCMs). Risk assessment methods are usually used in CPPI. However, risk evaluation is usually subjective. The purpose of this work is to present a more objective CPPI method. Methods: A CPPI method considering chemical composition, biological activity, and batch-to-batch con- sistency was presented in this work. The manufacturing process of notoginseng total saponins (NTS) was investigated as an example. The changes of chemical composition, biological activity, and chemical com- position consistency after main processes were measured and compared. A significant change of them indicated a critical process. Results: After extraction process and chromatography process, saponin purity and chemical composition similarity remarkably increased, and saponin content variations decreased. Thrombin inhibitory activity was remarkably decreased after chromatography process. Because of the large influences on NTS quality, extraction process and chromatography process were identified to be critical processes of NTS. Conclusion: Based on a comprehensive and objective examination of the role of each process, critical pharmaceutical processes can be identified. A similar method can also be applied to other TCM processes.

Mangiferin exerts neuroprotective activity against lead-induced toxicity and oxidative stress via Nrf2 pathway

2020/2/18 16:22:11

Objective: The study was designed to assess the beneficial role of mangiferin (MGN) in lead (Pb)-induced neurological damages in the activation of Nrf2-governed enzymes, genes and proteins. Methods: A total of 96 weaned Wistar rats (48 males and 48 females, 26- to 27-day-old), weighing 50?80 g were used. The experiment was performed in six groups: normal group (control, n = 16), model group (chronic Pb exposed, n = 16), Dimercaptosuccinic acid (DMSA)-treated group (positive control, Pb + DMSA, n = 16), three MGN-treated groups with different doses (Pb + MGN, n = 48). Normal group freely had access to purified water. DMSA-treated group was given DMSA, which was clinically used as the standard treatment for moderate Pb poisoning, at 50 mg/kg (2 mL suspension with purified water) by intragastric gavage (ig) 4 continual days a week for 4 weeks, MGN-treated groups were given MGN at 50, 100, or 200 mg/kg (2 mL suspension with purified water) by ig daily for 4 weeks. At the end of the treatment, all rats were sacrificed and the brain samples were collected. The haematoxylin and eosin (H&E) staining was used for observation of histopathology. Commercial kit, real-time quantitative poly- merase chain reaction (RT-qPCR), Western-blot and immunohistochemistry (IHC) detection were used to detect the mRNA and protein expression. Results: Eight weeks exposure to Pb-containing water resulted in pathological alterations, anti-oxidative system disorder in the brain, all of which were blocked by MGN in a Nrf2-dependent manner. Nrf2 down- stream enzymes such as HO-1, NQO1, γ -GCS were activated. Nrf2, GCLC, GCLM, HO-1 mRNA and total Nrf2, Nuclear Nrf2, γ -GCS, HO-1 protein expression were affected too. Conclusion: MGN ameliorated morphological damage in the hippocampus. Its neuroprotective effects were achieved by the activation of the Nrf2 downstream genes. The data from this in vitro study indi- cates that MGN targeting Nrf2 activation is a feasible approach to reduce adverse health effects associated with Pb exposure. Thus, MGN could be an effective candidate agent for the Pb-induced oxidative stress and neurotoxicity in the human body.

Phytochemical and anti-neuropathic investigations of Crocus sativus via alleviating inflammation, oxidative stress and pancreatic beta-cells regeneration

Karim Raafat, Maha Aboul-Ela, Abdalla El-Lakany — 2020/2/18 16:22:11

Objective: The aim of this study is to investigate the phytochemical and the long-term anti-neuropathic potentials of Crocus sativus cultivated in the University botanical garden, and explore its most bioactive compounds and their underlying mechanisms of action. Methods: Phytochemical analysis and bio-guided isolation-procedures including RP-HPLC and 1 H and 13 C NMR utilizing biological models of diabetes, inflammation, and diabetic-neuropathy were used. Cultivated saffron (S-RCED) and Spanish-saffron stigma (S-SP) alone or in combination with Camellia sinus (CS) were investigated. Results: The RP-HPLC analyses showed the presence of picrocrocin, crocin I, crocin II, crocin I’, crocin II’, and safranal (SAF) in both S-SP and S-RCED extracts with higher-concentrations. It had been shown that SAF was the most bioactive-compound in Crocus sativus. Both S-SP and S-RCED possessed significant (P < 0.05) anti-diabetic activities in acute (6 h), subchronic (8 d) and chronic (8 weeks) models. S-RCED had been proven with more hypoglycemic potentials when compared to S-SP and SAF. S-SP, S-RCED, and SAF produced significant anti-inflammatory and anti-nociceptive activities against carrageenan-induced inflammatory, hyperalgesic and tactile diabetic-neuropathy models, respectively. S-SP, S-RCED, and SAF elevated serum catalase, reduced glutathione, and insulin serum levels, ameliorated lipid peroxidation and HbA1c levels, and histopathologically regenerated the pancreatic beta-cells. Combinations with CS showed more significant efficacy than the single component. Conclusion: The oxidative stress reduction, insulin secretagogue, and pancreatic beta-cells regeneration potentials might be responsible for the mechanism underlying the anti-diabetic, anti-inflammatory and anti-diabetic neuropathy activities. Thus, the cultivated Crocus sativus might be clinically useful for pro- tecting against many serious-disorders.

Quantification of Chinese yam processing methods based on pyrolysis characteristics and its relation to Maillard reaction

2020/2/18 16:22:11

Objective: Chinese yam (Shanyao in Chinese, SY) as one of the representatives for Chinese medicines can be used as both of medicine and food with rich nutritional and medicinal value. Most of Chinese herbal medicines need to be processed prior to be used in clinical practice. SY was divided into Maoshanyao (Hairy Shanyao, MSY) and Guangshanyao (Smooth Shanyao, GSY) based on different processing meth- ods at the place of origin, and it also could be processed as stir-fried SY and bran stir-fried SY to meet the different clinical use. Moreover, during the processing of Chinese herbal medicines, more compli- cated Maillard reaction occurs compared to food processing. Therefore, the objective of this research is to quantify the firepower of SY processing, and combined this with the relevant parameters of Maillard reaction. Methods: The MSY and GSY produced in Shanxi and Henan Provinces were chosen as the research ob- jects. By using thermal analysis technology, we first established the correlation between pyrolysis and processing of SY and its mixtures. We also quantified the firepower of Shaoyao processing, and com- bined this with the relevant parameters of Maillard reaction (pH value, amino acid, and 5-HMF) and the changes in medicinal ingredients (allantoin). Results: The SY was mainly fried with moderate-fire (190 °C?200 °C), and the starting temperatures of different SY–ingredient mixtures were (176.3 ± 5.33) °C for (honey) bran, and (205.9 ± 8.05) °C for rice. The upper limits of processing temperature were (289.9 ± 6.47) °C for (honey) bran and (298.9 ± 1.15) °C for rice. The cooking time was (10.80 ± 1.76) min for soil stir-fry, (10.31 ± 1.06) min for bran stir-fry, and (8.43 ± 0.68) min for rice stir-fry. Moreover, the pH values and the content of 5-HMF were increased (P < 0.001), while the content of glycine was decreased significantly (P < 0.001) after processing. Conclusion: The results verified and quantified the firepower of traditional processing of SY, and also provided scientific reference for other studies related to SY processing.

Effects of total iridoid glycosides of Picrorhiza scrophulariiflora against non-alcoholic steatohepatitis rats induced by high-fat and high-sugar diet through regulation of lipid metabolism

2020/2/18 16:22:12

Objective: To investigate the therapeutic effect of total iridoid glycosides of Picrorhiza scrophulariiflora (TIGP) on non-alcoholic steatohepatitis (NASH). Methods: SD rats were fed with high-fat and high-sugar diet for 8 weeks to establish NASH. TIGP were given orally at doses of 20, 40 and 80 mg/kg/d for 4 weeks. Triglycerides assay (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspar- tate aminotransferase (AST), alanine aminotransferase (ALT), fasting plasma glucose (FPG), fasting insulin (FINS), tumor necrosis factor- α (TNF- α ), interleukin-6 (IL-6), chemokine-1 (MCP-1), leptin (LEP) in serum were tested. TG, TC, superoxide dismutase (SOD), malondialdehyde (MDA), and free fatty acid (FFA) in liver tissue were determined by colorimetric methods. Steatosis of hepatocytes and inflammation was performed by pathological examination. Results: The results showed that TIGP significantly decreased TC, TG and FFA in liver tissue, increased SOD activity, decreased MDA content, decreased serum levels of TG, TC, HDL-C/LDL-C, ALT, AST, GLU, HOMA-IR, TNF- α and LEP, and in addition, improved steatosis of liver cells compared to NASH. Conclusion: TIGP had anti-fatty liver effect against NASH rats induced by high-fat and high-sugar diet. Its mechanism was related to the regulation of lipid metabolism and reduction of insulin resistance, through inhibition of oxidative stress and inflammation.

Attenuation of inflammation in streptozotocin-induced diabetic rabbits by Matricaria chamomilla oil: A focus on targeting NF- κ B and NLRP3 signaling pathways

2020/2/18 16:22:12

Objectives: The present study was conducted to investigate the protective effects of chamomile oil from Matricaria chamomilla against type 1 diabetes mellitus (T1DM) and its potential mechanisms. Methods: T1DM was established in male New Zealand white rabbits via a single intraperitoneal infusion of streptozotocin (STZ) (80 mg/kg body weight ?1 , dissolved in 0.2 mL of normal saline). Different doses of chamomile oil (25, 50 and 100 mg/kg) were orally administrated to STZ induced diabetic rabbits for 21 consecutive days. The expression of pro-inflammatory cytokines was determined using ELISA assay. The expression of NF- κ B and NLRP3 was measured using Western blot assay. Results: Compared with normal rabbits, STZ-induced diabetic rabbits exhibited significant increased levels of blood glucose and decreased levels of serum insulin that were reversed using middle and high tested dose of chamomile oil. Likewise, STZ-induced diabetic rabbits showed a significant increased expression of NF- κ B and NLRP3 proteins in the pancreas tissue that was reversed by high tested dose of chamomile oil. Conclusion: Collectively, our findings demonstrated that chamomile oil possesses anti-hyperglycemic, and anti-inflammatory activities in STZ-induced diabetic rabbits by targeting inflammatory cytokines and NF- κ B and NLRP3 signaling pathways.

Pharmacokinetic studies of multi-bioactive components in rat plasma after oral administration of Xintiantai I extract and effects of guide drug borneol on pharmacokinetics

2020/2/18 16:22:12

Objective: To investigate the in vivo pharmacokinetic characteristics of 17 bioactive components includ- ing ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, dehydroevodiamine, rutaecarpine, limonin, hyperin, curcumin, demethoxycurcumin and bis- demethoxycurcumin in rat plasma after oral administration of Xintiantai I extract powder (XI) and Xin- tiantai I without guide drug borneol extract powder (XI without borneol), and study the compatibility effects of guide drug borneol on the pharmacokinetics. Methods: A UHPLC-MS/MS method was established and fully validated for the comparative pharmacoki- netics of 17 bioactive components. The pharmacokinetics parameters of 17 bioactive components after oral administration of XI and XI without borneol were calculated by the software of DAS 3.0 and inter- compared. Results: The specificity, linearity, lower limit of quantification (LLOQ), precision, accuracy, extraction re- covery rates, matrix effects, and stability of the UHPLC-MS/MS assay were good within the acceptance criteria from FDA guidelines. Guide drug borneol can significantly increase AUC of G-Rd, palmatine, hy- perin, curcumin, demethoxycurcumin, bisdemethoxycurcumin and C max of 16 bioactive components ex- cept for dehydroevodiamine (P < 0.05), decrease T max of G-Rd, berberine, columbamin, coptisine, limonin and MRT of 17 bioactive components in XI group (P < 0.05). Conclusion: Guide drug borneol enhanced the absorption of G-Rd, palmatine, hyperin, curcumin, demethoxycurcumin and bisdemethoxycurcumin.

A reliable and rapid pharmacokinetic study of pueraria isoflavones using pueraria reference extractive substance in beagle plasma: Application to study of Yufeng Ningxin Tablets

2020/2/18 16:22:13

Objective: In order to evaluate the reliability and feasibility of pueraria reference extractive substance (RES) used in biological sample, the pharmacokinetics of 3 ? –hydroxy puerarin (3 ? -HP), puerarin, 3 ? – methoxy puerarin (3 ? -MP), and daidzein-8-C-apiosyl-(1-6)-glucoside (DAG) in beagle plasma following oral administration of Yufeng Ningxin Tablet were quantitated. Methods: A reliable and sensitive high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS) method developed with chromatographic separation was oper- ated on a Merck C 18 column, and acetonitrile-5 mmol/L ammonium was used as mobile phase in gradient elution. The plasma samples were deproteinized by acetone, detected by triple quadrupole mass spec- trometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. The pharmacokinetic parameters were calculated by Winnonlin 4.1. Results: The calibration curves of the reference extractive substance and standard substance methods were linear over the ranges 0.0417–11.3309 μg/mL and 0.0394–10.0000 μg/mL. The intra-day and inter- day precision of the two methods at three concentrations were less than 13.63%, and the average recov- eries of 3 ? -HP, puerarin, 3 ? -MP, and DAG were more than 70.67%. The RSD of the mean plasma concentra- tions of the analytes calculated by the two methods was less than 5%, and cos (?) = =1.000. Among the analytes, puerarin showed the highest blood concentration [(940 ± 185) ng/mL] and the longest retention time [(5 ± 1) h] in the dog’s bodies. Conclusion: Pueraria reference extractive substance can be seen as an alternative to the standard sub- stance to overcome the scarcity of standard substance for the analysis of biological samples.

Analysis on oral medication rules of traditional Chinese medicine prescriptions for prevention of COVID-19

2021/11/17 13:46:22

Objective: To analyze the medication rules of traditional Chinese medicine (TCM) oral prescriptions for the prevention of COVID-19. Methods: The preventive oral prescriptions for COVID-19 published by national and provincial health and wellness committees, administrations of TCM, medical institutions at all levels, and medical masters and Chinese medicine experts were collected to establish a database, manual screening was carried out according to the inclusion and exclusion criteria, and frequency statistics, association rule analysis. The mutual information method, entropy hierarchical clustering and other methods were improved through Excel and the TCM inheritance auxiliary platform V2.5 to mine the rules and characteristics of medication. Results: The selected 157 prescriptions contained a total of 130 TCMs. The top five TCMs with the highest use frequency were Glycyrrhizae Radix et Rhizoma (86), Astragali Radix (80), Lonicerea Japonicae Flos (70), Atractylodis Macrocephalae Rhizoma (62), Saposhnikoviae Radix (60). In accordance with TCM efficacy classification, most of them were medicines for qi-tonifying (279), followed by medicines for clearing heat and drying dampness (163), dispelling pathogenic wind-cold (126), resolving dampness (111), as well as dispelling pathogenic wind-heat (99). The characteristics of four-natures of the selected medicines are as follows: most of them were cold (59), followed by warm (38) and mild (21). In terms of five-taste, most of them were sweet (26) and acrid-and-bitter (24), followed by sweet-and-bitter (20), bitter (20) and acrid (15). For the meridian attribution, the five-zang organs and six-fu organs were all involved, most of them attributed to lung meridian (80), followed by stomach meridian (57) and spleen meridian (40). Based on association rule analysis, 12 commonly used medicine combinations with two or three TCMs were found. The commonly used medicinal pairs included Astragali Radix and Saposhnikoviae Radix (51), Astragali Radix and Atractylodis Macrocephalae Rhizoma (46), Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix (43), Astragali Radix and Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix (38), Forsythiae Fructus and Astragali Radix (37), and so on. In addition, 14 core combinations of medicines were obtained by complex system entropy cluster analysis, on this basis, six new prescriptions were screened out based on unsupervised entropy hierarchical clustering analysis. According to The Catalogue of Edible Traditional?Chinese?Medicinal?Materials, Traditional?Chinese?Medicinal?Materials for Health Food, and New Resources of Food published by National Health Commission of the People’s Republic of China, there are 35 species belonging to the group of edible traditional?Chinese?medicinal?materials, 20 species belonging to the group of new resources of food, 31 species belonging to the group of traditional?Chinese?medicinal?materials for health food, 19.11% of the preventive oral prescriptions for COVID-19 were composed of the medicines belonging to the above three groups. Besides, there are 11 toxic species, and 24.84% of the preventive oral prescriptions for COVID-19 contained toxic traditional Chinese medicines. Conclusion: We found that invigorating qi and resolving dampness were the main treatment used to prevent for COVID-19, combined with the methods for strengthening vital energy and eliminating pathogenic factors. Most of the preventive oral prescriptions for COVID-19 were treated in lung, spleen and stomach meridians. In the process of selecting prescriptions and using TCMs to prevent for COVID-19, the safety of preventive medicines was also emphasized. And the theory of "Preventive Treatment of Disease" was embodied in these preventive oral prescriptions for COVID-19. For the prescriptions containing toxic TCMs, special?attention?should be?paid?to their safety in clinical application.

Analysis on application law of dampness-removing Chinese medicine in treatment of coronavirus disease 2019

2021/11/17 13:46:22

Objective: Coronavirus Disease 2019 (COVID-19) has been diagnosed as an epidemic disease characterized by cold and dampness pathogens in TCM clinic. Due to many Chinese medicines with different functions were used in the treatment of COVID-19, it is very important to find the law of application of damp-removing Chinese medicine with high frequency application, with view to providing a reference for the use and research of Chinese medicine to further control the pandemic. Methods: The publicly released diagnosis and treatment programs issued by the National Health Commission and Health Commission of provinces, autonomous regions and municipalities, and Chinese herbs prescription information in these were collected, a database was established, and Excel and Graphpad 8.0 software were used to analyze the frequency of use of various Chinese medicines, the frequency and property characters including five flavors (bitter, pungent, sweet, sour, and salty) and four qi (warm, hot, cool, and cold) and channel tropisms of Chinese medicine for removing dampness. Results: A total of 137 prescriptions of Chinese medicine for treating COVID-19 were collected, including 178 TCMs showing functions of resolving phlegm, relieving cough and asthma, resolving dampness, clearing damp and inducing dieresis, clearing heat, tonifying deficiency, and relieving exterior syndrome, in which the TCMs with the first four functions that we called the removing-dampness TCMs, accounted for 35.78%. Also a number of TCMs in the rest functions showed removing-dampness. The first four functions were divided into subfunctions including aromatic resolving?dampness, clearing heat and drying dampness, drying dampness and tonifying spleen qi, drying dampness and removing phlegm, inducing diuresis and relieving swelling, inducing diuresis and relieving exterior syndrome. Among them, the most frequently used TCMs was Ephedrae Herba, followed by Citri Reticulatae Pericarpium, Pogostemonis Herba, Pinelliae Rhizoma, Poria, Scutellariae Radix, and Atractylodis Rhizoma. The property character analysis in the removing-dampness TCMs showed that bitter and pungent were largely present and sour and astringent were absent, warm and hot were dominant; And the lung, spleen, stomach, large intestine, bladder were main channel tropisms. Conclusion: This study may provide meaningful and useful information on further research to investigate the effective compounds from the dampness-removing Chinese medicine with high frequency application, and also provide a reference for the clinical treatment of COVID-19 accurately against dampness evil with dampness-removing Chinese medicine.

Common mechanism of Citrus Grandis Exocarpium in treatment of chronic obstructive pulmonary disease and lung cancer

2021/11/17 13:46:22

Objective: “Same treatment for different diseases” is a unique treatment strategy in traditional Chinese medicine. Two kinds of malignant respiratory diseases endanger human health-chronic obstructive pulmonary disease (COPD) and lung cancer. Citrus Grandis Exocarpium (Huajuhong in Chinese, HJH), a famous herbal, is always applied by Chinese medicine practitioners to dispersion the lung toresolve phlegm based on “syndrome differentiation and treatment” theory. However, the common mechanism for HJH’s treatment of COPD and lung cancer is not clear. Methods: In this study, based on network pharmacology and molecular docking technology, the common mechanism of HJH in the treatment of COPD and lung cancer was studied. The active ingredients and related targets of HJH were integrated from TCMSP, BATMAN-TAM, STP, and Pubchem databases. The standard names of these targets were united by UniProt database. Targets of COPD and lung cancer were enriched through GeneCards, NCBI (Gene), Therapeutic Target Database, and DisGeNET (v7.0) databases. Then the intersection targets of HJH and diseases were obtained. The STRING network and the Cytoscape 3.7.2 were used to construct PPI network, the DAVID database was used to perform GO and KEGG analysis. Then Cytoscape 3.6.1 was used to build “ingredient-target-signal pathway” network. Finally, AutoDock 1.5.6 software was used to perform molecular docking of key proteins and molecules. Results: Eleven active ingredients in HJH were obtained by searching the database, corresponding to 184 HJH-COPD-lung cancer targets intersection. The results of biological network analysis showed that naringenin, the active component in HJH, could mainly act on target proteins such as AKT1, EGFR. Then through positive regulation of vasoconstriction and other biological processes, naringenin could regulate estrogen signaling pathway, VEGF signaling pathway, HIF-1 signaling pathway, ErbB signaling pathway, PI3K-Akt signaling pathway to play an important role in the treatment of both COPD and lung cancer. Conclusion: Network pharmacology was employed to systematically investigate the active ingredients and targets of HJH in treatment of COPD and lung cancer. And then, the common pharmacodynamic network of HJH for the two malignant respiratory diseases was firstly described. Furthermore, naringenin was proved to strongly bind with AKT1 and EGFR. It may provide the scientific basis for understanding the “Same treatment for different diseases” strategy in traditional Chinese medicine and inspirit subsequent drug discovery for COPD, lung cancer and other malignant lung diseases.

High water-soluble curcuminoids-rich extract regulates osteogenic differentiation of MC3T3-E1 cells: Involvement of Wnt/Î²-catenin and BMP signaling pathway

2021/11/17 13:46:22

Objective The present study aimed to evaluate the effect of a high water-soluble curcuminoids-rich extract (CRE) in a solid dispersion form (CRE-SD) using polyvinylpyrrolidone K30 on osteogenic induction of MC3T3-E1 cells. Methods CRE was pre-purified using a microwave assisted extraction couple with a Diaion? HP-20 column chromatography. The osteoblastic cell proliferation and differentiation potentials of CRE-SD in MC3T3-E1 cells were tested by cell viability, alkaline phosphatase (ALP) activity, and Alizarin red S activity assays. The mRNA expressions of osteoblast-specific genes and underline mechanisms were assessed by a real time PCR and western blot analysis. Results CRE-SD 50 μg/mL increased alkaline phosphatase (ALP) activity, an early differentiation marker of osteoblasts in both MC3T3-E1 cells and non-osteogenic mouse pluripotent cell line, C3H10T1/2, indicating the action of CRE-SD was not cell-type specific. Alizarin red S activity showed a significant amount of calcium deposition in cells treated with CRE-SD. CRE-SD also upregulated the mRNA expression levels of transcription factors that favor osteoblast differentiation including Bmp-2, Runx2 and Collagen 1a, in a dose dependent manner. Western blot analysis revealed that noggin attenuated CRE-SD-promoted expressions of Bmp-2 and Runx2 proteins. siRNA mediated blocking of Wnt/β-catenin signaling pathway also annulled the influence of CRE-SD, indicating Wnt/β-catenin dependent activity. Inhibition of the different signaling pathways abolished the influence of CRE-SD on ALP activity, confirming that CRE-SD induced MC3T3-E1 cells into osteoblasts through Wnt/β-catenin and BMP signaling pathway. Conclusion These results collectively demonstrate that CRE-SD may be a potential therapeutic agent for the treatment of osteoporosis.

In vitro and in vivo antihypertensive and antioxidant activities of fermented roots of Allium hookeri

Tao Tonga, Yanan Wanga, Cheng-Mei Zhangb, Seong-Gook Kangc — 2021/11/17 13:46:22

Objective: To evaluate antihypertensive and antioxidant activities of Allium hookeri root (AHR) fermented with Lactobacillus plantarum, Leuconostoc mesenteroides, and Weissella cibaria. Methods: The novel fermented AHR products using L. plantarum, L. mesenteroides, and W. cibaria were developed and ACE inhibitory activity, DPPH radical scavenging activity, hydroxyl radical scavenging activity, superoxide anion radical scavenging activity, total phenolic content, and total thiosulfinate content were determined. The antihypertensive and antioxidant effects of fermented AHR were further investigated in spontaneously hypertensive rats (SHRs). Results: Administration of fermented AHR to SHRs had an attenuating effect on both diastolic and systolic blood pressure. The SHRs treated with fermented AHR showed lower plasma ACE activity and higher plasma NO levels. Furthermore, fermented AHR administration led to parallel improvements in plasma oxidative stress biomarkers in SHRs. Conclusion: Our results highlight the potential usefulness of fermented AHR for the prevention of hypertension.

Combined enzymatic hydrolysis and herbal extracts fortification to boost in vitro antioxidant activity of edible birdâ€™s nest solution

Pei Ling Tanga, Hooi Shin Goha, Swee Seng Siab — 2021/11/17 13:46:22

Objective Edible bird’s nest (EBN) is a popular traditional tonic food in Chinese population for centuries. Malaysia is one of the main EBN suppliers in the world. This study aims to explore the best strategy to boost the antioxidant potential of EBN solution. Methods In this study, the raw EBN (4%, mass to volume ratio) was initially enzymatic hydrolyzed using papain enzyme to produce EBN hydrolysate (EBNH), then spray-dried into powdered form. Next, 4% (mass to volume ratio) of EBNH powder was dissolved in ginger extract (GE), mulberry leaf extract (MLE) and cinnamon twig extract (CTE) to detect the changes of antioxidant activities, respectively. Results Results obtained suggest that enzymatic hydrolysis significantly reduced the viscosity of 4% EBN solution from (68.12 ± 0.69) mPa.s to (7.84 ± 0.31) mPa.s. Besides, the total phenolic content (TPC), total flavonoid content (TFC), total soluble protein, DPPH scavenging activity and ferric reducing antioxidant power (FRAP) were substantially increased following EBN hydrolysis using papain enzyme. In addition, fortification with GE, MLE and CTE had further improved the TPC, TFC, DPPH scavenging activity and FRAP of the EBNH solution. Among the samples, MLE-EBNH solution showed the most superior antioxidant potential at (86.39 ± 1.66)% of DPPH scavenging activity and (19.79 ± 2.96) mmol/L FeSO4 of FRAP. Conclusion This study proved that combined enzymatic hydrolysis and MLE fortification is the best strategy to produce EBN product with prominent in vitro antioxidant potential. This preliminary study provides new insight into the compatibility of EBN with different herbal extracts for future health food production.

Qualitative and quantitative analysis of Arnebiae Radix and Dictamni Cortex and efficacy study of herbal extracts on allergic contact dermatitis using 3D human reconstructed epidermis

2021/11/17 13:46:22

Objective: To evaluate the quality of Arnebiae Radix (AR) and Dictamni Cortex (DC) and study the efficacy of herbal extracts of these two herbs on the treatment of allergic contact dermatitis (ACD). Methods: Qualitative and quantitative analysis of effective components was performed using High Performance Thin Layer Chromatography (HPTLC), High Performance Liquid Chromatography (HPLC), and HPLC-Quadrupole Time of Flight-Mass Spectrometry (HPLC-QTOF-MS). In vitro allergic ACD 3D model was established by incubating 3D reconstructed human epidermis (RHE) with skin sensitizer, potassium dichromate. A total of 65 gene expression that were associated with ACD, which included 24 antioxidant responsive element (ARE) and 41 SENS-IS genes were quantified by qRT-PCR. More than or equal to 10 ARE genes and 18 SENN-IS genes were induced by 1.3-fold, demonstrating the successful establishment of in vitro ACD model. Oil extracts of AR and DC were applied on the in vitro ACD model to study the efficacy. Results: Batch 3 of AR and batch 2 of DC showed presence of all active ingredients with the highest concentrations. Active ingredients of the herbs were extracted using a special oil and formulated into herbal oil extracts. The herbal oil extracts were able to down regulate the induced genes in the in-vitro ACD skin model, bringing the tissue back to homeostatic status. Conclusion: The oil extracts showed the potent efficacy of using AR and DC in ACD treatment. The combination study will be done to optimize the formulation ratio which will be developed into a topical cream.

Submerged fermentation improves bioactivity of mulberry fruits and leaves

Hui Qian Chuah, Pei Ling Tang, Ni Jing Ang, Hui Yin Tan — 2021/11/17 13:46:22

Objective Mulberry (Morus spp.) fruits and leaves have been proven to possess nutraceutical properties. Due to its fast and easy growing characteristics, mulberry fruits (MF) and leaves (ML) potentially emerge as a great source of functional foods. This study aims to enhance bioactivities (antioxidant, anti-inflammation, and hypoglycemic activity) of MF and ML via submerged fermentation using bacteria (Lactobacillus plantarum TAR 4), yeast (Baker’s yeast and red yeast) and fungi (Tempeh and Tapai starter). Methods In this study, 25% (mass to volume ratio) of MF and ML were fermented (48 h) with 1% (mass to volume ratio) of different microbial cultures, respectively. Effects of different fermentations on MF and ML were determined based on the changes of total phenolics (TPC), flavonoids (TFC), anthocyanins, total sugar, DPPH activity, ferric reducing antioxidant power (FRAP), albumin denaturation inhibition activity (ADI), anti-lipoxygenase activity and α-amylase inhibition activity (AI). Results Generally, ML had higher AI than MF. However, MF exhibited higher DPPH, FRAP and anti-lipoxygenase activity than ML. After all forms of fermentation, DPPH and AI activity of MF and ML were increased significantly (P < 0.05). However, the effects of fermentation on TPC, FRAP, ADI and anti-lipoxygenase activity of MF were in contrast with ML. TPC, FRAP and anti-lipoxygenase activity of ML were enhanced, but reduced in MF after fermentation. Although the effects exerted by different microorganisms in MF and ML fermentation were different, the bioactivities of MF and ML were generally improved after fermentation. Fermentation by Tempeh starter enhanced TPC (by 2-fold), FRAP (by 2.3-fold), AI (at 10% increment) and anti-lipoxygenase activity (by 5-fold) of ML, whereas Tapai fermentation effectively enhanced the DPPH (at 17% increment) and ADI (by 2-fold) activity of MF. Conclusion Findings of this study provide an insight into the future process design of MF and ML processing into novel functional foods.

Systems pharmacology dissection of action mechanisms for herbs in osteoporosis treatment

2021/8/2 9:37:07

Objective: Osteoporosis has become the biggest cause of non-fatal health issue. Currently, the limitations of traditional anti-osteoporosis drugs such as long-term ill-effects and drug resistance, have raised concerns toward complementary and alternative therapies, particularly herbal medicines and their natural active compounds. Thus, this study aimed to provide an integrative analysis of active chemicals, drug targets and interacting pathways of the herbs for osteoporosis treatment. Methods: Here, we introduced a systematic pharmacology model, combining the absorption, distribution, metabolism, and excretion (ADME) screening model, drug targeting and network pharmacology, to probe into the therapeutic mechanisms of herbs in osteoporosis. Results: We obtained 86 natural compounds with favorable pharmacokinetic profiles and their 58 targets from seven osteoporosis-related herbs. Network analysis revealed that they probably synergistically work through multiple mechanisms, such as suppressing inflammatory response, maintaining bone metabolism or improving organism immunity, to benefit patients with osteoporosis. Furthermore, experimental results showed that all the five compounds (calycosin, asperosaponin VI, hederagenin, betulinic acid and luteolin) enhanced osteoblast proliferation and differentiation in vitro, which corroborated the validity of this system pharmacology approach. Notably, gentisin and aureusidin among the identified compounds were first predicted to be associated with osteoporosis. Conclusion: Herbs and their natural compounds, being characterized as the classical combination therapies, might be engaged in multiple mechanisms to coordinately improve the osteoporosis symptoms. This work may contribute to offer novel strategies and clues for the therapy and drug discovery of osteoporosis and other complex diseases.

Comparison of anti-inflammatory effects of Lonicerae Japonicae Flos and Lonicerae Flos based on network pharmacology

Yu Gao, Feng-xue Wang, Qing Liu, Yao-dong Qi, Qiu-ling Wang, Hai-bo Liu* — 2021/8/2 9:37:07

Objective In TCM history, Lonicerae Japonicae Flos and Lonicerae Flos were used clinically as one drug, but now they are admitted as two herbal medicines in Chinese Pharmacopoeia (2010 edition). This study used network pharmacology to investigate whether the two can be used interchangeably for the treatment of inflammatory diseases in TCM clinical practice. Methods Lonicerae Japonicae Flos and Lonicerae Flos were compared in the inflammation mechanism including core targets, Gene Ontology (GO), pathway and principle chemical components by the method of network pharmacology. Results Both herbs shared in six targets accounting for 66.7% of the entire core targets and more than half of the GO terms and pathways are similar. Organic acids are dominent compounds responsible for anti-inflammatory effects. Three of the compounds that bind to core targets including luteolin, quercetin and kaempferol, are shared in both herbs. Conclusion Due to high similarity between Lonicerae Japonicae Flos and Lonicerae Flos, we believe that they can be used interchangeably for the inflammation in clinical treatment.

Investigating pharmacological mechanisms of andrographolide on non-alcoholic steatohepatitis (NASH): A bioinformatics approach of network pharmacology

Lei Li a,b, Sheng-he Li a, Jin-peng Jiang a, Chang Liu a, *, Li-li Ji b,* — 2021/8/2 9:37:07

Objective: To investigate the mechanisms of andrographolide against non-alcoholic steatohepatitis (NASH) based on network pharmacology, so as to provide a reference for further study of andrographolide in the treatment of NASH and other metabolic diseases. Methods: The methionine- and choline-deficient (MCD) diet-induced NASH mice were treated by administration of andrographolide, and serum transaminase and pathological changes were analyzed. The network pharmacology-based bioinformatic strategy was then used to search the potential targets, construct protein-protein interaction (PPI) network, analyze gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment, and conduct molecular docking to explore the molecular mechanisms. Results: The predicted core targets TNF, MAPK8, IL6, IL1B and AKT1 were enriched in non-alcoholic fatty liver disease (NAFLD) signaling pathway and against NASH by regulation of de novo fatty acids synthesis, anti-inflammation and anti-oxidation. Conclusion: This work provides a scientific basis for further demonstration of the anti-NASH mechanisms of andrographolide.

Mechanism of Huoxue Tongluo Decoction in treatment of erectile dysfunction caused by ischemic stroke based on network pharmacology

2021/8/2 9:37:07

Objective: To study the therapeutic effect of Huoxue Tongluo Decoction (HXTLD) on erectile dysfunction caused by ischemic stroke and identify the mechanisms involved. Methods: Network pharmacology was used to predict the key active ingredients and targets of HXTLD. Surgical methods were used to create a rat model of ischemic stroke. The rats were then given a suspension of HXTLD by ig administration. Erectile function was evaluated by Apomorphine (APO) induction. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (Real-time PCR) and Western blotting were used to detect the expression of related mRNAs and proteins in rat penile corpus cavernous tissue and brain tissue. Hematoxylin & Eosin (HE) staining was used to investigate structural changes in the penile cavernous tissue. Results: Network pharmacology showed that tumor necrosis factor (TNF), nitric oxide synthase 3 (eNOS), and vascular endothelial growth factor (VEGF) were the key targets of HXTLD in the treatment of erectile dysfunction caused by ischemic stroke. Experimental studies showed that HXTLD improved erectile dysfunction caused by ischemic stroke. HE results showed that HXTLD improved the structure of the corpus cavernosa. HXTLD also inhibited the expression of TNF and VEGF proteins in penile tissue (P < 0.05) and enhanced the expression of eNOS protein in penile tissue (P < 0.05). Conclusion: HXTLD improved the erectile function of rats with erectile dysfunction caused by ischemic stroke by regulating the mRNA and protein levels of TNF, eNOS and VEGF.

Dual inhibition of COVID-19 spike glycoprotein and main protease 3CLpro by Withanone from Withania somnifera

2021/8/2 9:37:08

Objective To identify the safe and effective natural inhibitors of spike glycoprotein and main protease 3CLpro using potential natural antiviral compounds which are studied under various animal models and viral cell lines. Methods First, compounds were retrieved from the PubChem database and predicted for their druggability using the MolSoft web server, and compounds having drug-like property were predicted for major adverse drug reactions like cardiotoxicity, hepatotoxicity, arrhythmia, myocardial infarction, and nephrotoxicity using ADVERpred. Docking of nontoxic antiviral compounds with spike glycoprotein and main protease 3CLpro was performed using AutoDock vina by PyRx 0.8 version. The stability of compound-protein interactions was checked by molecular dynamic (MD) simulation using Schrodinger Desmond software. Results Based on the druggable and nontoxic profile, nine compounds were selected. Among them, Withanone from Withania somnifera showed the highest binding affinity and best fit at active sites 1 of spike glycoprotein (glycosylation site) and main protease 3CLpro via interacting with active site amino acid residues before and after MD simulation at 50 ns. Withanone, which may reduce the glycosylation of SARS-CoV-2 via interacting with Asn343 and inhibit viral replication. Conclusion The current study reports Withanone as a non-toxic antiviral against SARS-CoV-2 and serve as a potential lead hit for further experimental validation.

Self-assembled aggregations in Coptis chinensis decoction dynamically regulate intestinal tissue permeability through Peyer's patch-associated immunity

2021/8/2 9:37:08

Objective To investigate the dynamic regulation of self-assembled aggregations (SAA) in Coptis chinensis (C. chinensis) decoction on the permeability of intestinal tissue and the mechanism underlying. Methods The effects of SAA on berberine (Ber) absorption were respectively analyzed in an in situ intestinal perfusion model and in an Ussing Chamber jejunum model with or without Peyer's patches (PPs). The expression levels of ZO-1, Occludin and Claudin-1 were detected by immunofluorescence to evaluate the tight junction (TJ) between intestinal epithelium cells. The expression levels of T-box-containing protein expressed in T cells, signal transducers and activators of tranion-6, retinoic acid receptor-related orphan receptor γt and forkhead box P3 in PPs were detected by the reverse transcription-polymerase chain reaction and the secretions of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-β (TGF-β) in PPs were evaluated by immunohistochemistry, to reflect the differentiation of T lymphocyte in PPs to helper T (Th) cell 1, Th2, Th17 and regulatory T (Treg) cell. To confirm the correlation between SAA in C. chinensis decoction, PPs-associated immunity and intestinal epithelium permeability, SAA were administrated on an Ussing Chamber jejunum model with immunosuppressed PPs and evaluated its influences on intestinal tissue permeability and TJ proteins expression. Results SAA in C. chinensis decoction could dose-dependently promote Ber absorption in jejunum segment, with the participation of PPs. The dose-dependent and dynamical regulations of SAA on permeability of intestinal tissue and TJ proteins expression level between intestinal epithelium cells occurred along with the dynamically changed T lymphocyte differentiation and immune effectors secretion in PPs. The administration of SAA on immunosuppressed PPs exhibited dose-dependent PPs activation, inducing dynamic promotion on intestinal tissue permeability and inhibition on TJ proteins expression. Conclusion SAA can improve the Ber absorption in small intestine, through the PPs-associated immunity induced dynamic regulation on intestinal tissue permeability and TJ proteins expression. These findings might enlighten the research of traditional Chinese medicine decoction.

Polyphenols from Securidaca inappendiculata alleviated acute lung injury in rats by inhibiting oxidative stress sensitive pathways

2021/8/2 9:37:08

Objective: Securidaca inappendiculata is a medicinal plant frequently used in the treatment of inflammatory diseases in south China. In this study, we aimed to explore its bioactive constituent which contributes to the anti-inflammatory activity. Methods: Polyphenol-enriched and polyphenol-deprived fractions (PRF and PDF, respectively) were separated from the ethanolic extract by HPD300 macroporous resin-based method, and their anti-inflammatory activities were investigated on a lipopolysaccharide (LPS)-induced acute lung injury (ALI) model in rats. The possible mechanism of action in alleviating acute inflammation was studied using RAW264.7 cells. Results: Both Folin-Ciocalteu and 1H nuclear magnetic resonance (NMR) analyses showed that polyphenolic content in PRF was approximately 10 times higher than that of PDF, and this observation reflected in their antioxidative capacities. PRF but not PDF significantly decreased the level of malondialdehyde, suppressed the expression of nicotinamide phosphoribosyltransferase (NAMPT) protein, and improved the severity of ALI in rats. PRF at 10 μg/mL effectively downregulated the expression of proteins NAMPT, HMGB1, TLR4, and p-p65, and scavenged the intracellular reactive oxygen species (ROS) in LPS- primed RAW264.7 cells. N-acetyl-L-cysteine exhibited similar inhibitory effects on ROS production and NAMPT-mediated TLR4/NF-κB activation in vitro, whereas nicotinamide mononucleotide antagonized all the changes induced by PRF during cotreatments. Conclusion: As an antioxidant, PRF exhibited potent anti-inflammatory activity under both in vivo and in vitro conditions by downregulating NAMPT and TLR4/NF-κB. Accordingly, polyphenols were identified as important bioactive constituents in S. inappendiculata targeting oxidative stress-sensitive pro-inflammatory pathways.

Discovery and identification of quality markers of Sparganii Rhizoma based on zebrafish thrombosis model

Nan Xu1, Rong Sun2, Yin-ping Shi3, Li-wen Han4, Hai-yan Shi3,* — 2021/8/2 9:37:08

Objective: The aim of the present study was to determine the Q-markers of Sparganii Rhizoma against thrombus through an integration of investigations on its antithrombotic effect, content determination and spectrum-effect correlation analysis. Methods: Based on the concept of Q-Marker, Sparganii Rhizoma was investigated for the identification of chemical component. The pharmacological effects on arachidonic acid-induced thrombosis in zebrafish were also investigated. The material basis in ethanol extract was determined by HPLC-UV. Furthermore, the potential Q-markers were analyzed and predicted according to the effect-chemical correlation analysis. Finally, the anti-thrombotic Q-markers were verified through the anti-thrombotic test of monomer components. Results: The model of thrombosis zebrafish was established with larvae exposed to 100 μmol/L arachidonic acid for 1 h. Nine ingredients in Sparganii Rhizoma were identified as 5-hydroxymethylfurfural, vanillic acid, ferulic acid, p-hydroxybenzaldehyde, p-hydroxybenzoic acid, vanillin, protocatechuic acid, p-coumaric acid and isoferulic acid. According to the determination effect of zebrafish thrombosis model and HPLC content analysis results, all the other contents present positive correlation except 5-hydroxymethylfurfural, and the P values of three representative potential Q-markers (ferulic acid, protocatechuic acid and p-coumaric acid) were 0.002, 0.001 and 0.026, respectively. Conclusion: Sparganii Rhizoma showed a dose-dependent effect on the recovery of reducing cardiac red blood cell on zebrafish model. Three phenolic acids (ferulic acid, protocatechuic acid and p-coumaric acid) were proved to possess the anti-thrombotic effects which could be regarded as the potential Q-markers for quality assessment of Sparganii Rhizoma.

Molecular mechanism of Chuanxiong Rhizoma in treating coronary artery diseases

2021/8/2 9:37:08

Objective: Most of the studies on the herb Chuanxiong Rhizoma (CR) have focused on the l-arginine-nitric oxide (NO) pathway, but the nitrate-nitrite-NO (NO3?-NO2?-NO) pathway was rarely investigated. Therefore, the aim of this study was to evaluate the effects and mechanisms of action of CR in coronary artery disease (CAD). Methods: The NO3?, NO2? and NO levels were examined in the NO3?-NO2?-NO pathway. High-performance ion chromatography was used to quantify NO3? and NO2? levels. Then, NO was quantified using a multifunctional enzyme marker with a fluorescent probe. The tension of aortic rings was measured using a multi myograph system. Results: High content of NO3? and low content of NO2? was found in CR, and which could potently convert NO3? to NO2? in the presence of endogenous reductase enzyme. Incubating human coronary artery endothelial cells (HCAECs) with CR-containing serum showed that CR significantly decreased the NO3? content and increased the levels of NO2? and NO in the cells under hypoxic conditions. In addition, CR significantly relaxed isolated aortic rings when the l-arginine -NO pathway was blocked. The optimal concentration of CR for relaxation was 200 mg/mL. Conclusion: CR supplements large amounts of NO in cells and vessels to achieve relaxation via the NO3?-NO2?-NO pathway, thereby making up for the deficiency caused by the lack of NO after the l-arginine-NO pathway is suppressed. This study also supports the potential use of a traditional Chinese herb for future drug development.

Anaphylaxis effect and substance basis of honeysuckle extract

Yu-fei Fenga, Guo-zhao Qina, Zhong-xu Jingb, Yan-hong Wanga, Yan-yan Zhoua,* — 2021/8/2 9:37:08

Objective: To explore the anaphylaxis effect and anaphylaxis substances of honeysuckle. Methods: Rat peritoneal mast cells (PMC) were separated and purified, the cells were incubated with compound 48/80 (0.02 g/L), physiological saline and honeysuckle extract (120 g/L) at 37 °C for 0, 15, 30, 45 and 60 min. Degranulation were observed by optical microscope and transmission electron microscope. Annexin?V?positive?cell?rate was detected by?flow cytometry to reflect the degranulation rate of PMC. SD rats were supplied with honeysuckle extract by?intravenous?injection at a dose of 2.25 g/L. After administration, different parameters were analyzed, including the symptoms, histamine (HIS) and tryptase (MCT) levels, which were determined to explore the effect of anaphylaxis. Regression analysis was used to calculate the relationships between the peaks and the pharmacological effects to explore potentially anaphylactoid components. Results: The percentage of Annxin V positive cells and the degranulation ratio were markedly elevated in PMC treated with honeysuckle extract for more than 15 min (P < 0.05). HIS and MCT level were significantly elevated after injection of honeysuckle extract for more than 15 min. Morphology of PMC and systemic symptoms were also changed compared with the controlled group (P < 0.05). Regression analysis was used to calculate the relationship between peaks and pharmacological effects, and to determine peaks 7, 10 and 13 as possible anaphylactoid ingredients. Conclusion: This study established a prospective method to clarify the anaphylactoid components of honeysuckle extract, which would provide guidance for screening anaphylactoid components in traditional Chinese medicine injections containing honeysuckle in the prescription.

Ethanol extract of Ardisiae Japonicae Herba inhibits hepatoma carcinoma cell proliferation in vitro through regulating lipid metabolism

2021/8/2 9:37:08

Objective: The aim of this study is to discover the possible working mechanisms of Ardisiae Japonicae Herba (AJH) on hepatoma carcinoma (HCC). Methods: In this study, ethanol extract of AJH was prepared and used to treat HCC cell in vitro. Furthermore, a genomic wide RNA sequencing (RNA-seq) was performed to screen deregulated genes in HCC cells after the treatment of AJH extract. The gene and protein expression related to lipid metabolism in HCC cells were also investigated to validate the results obtained from RNA-seq. Results: AJH extract could inhibit HCC cell proliferation in vitro. RNA-seq analysis has identified 1,601 differentially expressed genes (DEGs, fold change ≥ 2.0 or fold change ≤ 0.5, P < 0.05) in HCC after AJH extract treatment, which included 225 up-regulated genes and 1,376 down-regulated genes. KEGG pathway analysis of DEGs demonstrated that lipid metabolism was a potential pathway related to AJH treatment. In agreement with the RNA-seq data, qPCR and Western-blot analysis indicated that expression of genes and proteins related to lipid metabolism (SREBP1, ACC, ACLY and FASN) were significantly down-regulated in AJH treatment group as compared with the control group. Furthermore, AJH extract could also decrease lipid contents and cellular free fatty acid levels in HCC cells. Conclusion: Ethanol extract of AJH could inhibit HCC cell proliferation in vitro, the possible mechanism may be related to the inhibition of lipid metabolism.

Genome sizes of four important medicinal species in Kadsura by flow cytometry

2021/8/2 9:37:08

Objective: Dianjixueteng is a geoherb in Yunnan Province, the source plant of which is Kadsura interior. However, the formation of this geoherb is not clear in genetic mechanism, in which genome size is the first step that should be known on the genomic level. In this study we aimed to estimate the genome sizes of source plants of K. interior compared with three related herbs K. heteroclita, K. longipedunculata, and K. coccinea by flow cytometry (FCM). Methods: The genome sizes of K. interior, K. heteroclita, K. longipedunculata and K. coccinea, i.e., the source plants of K. interior and its relative medicinal materials, were estimated by FCM. The nuclei of K. interior were isolated using modified LB01 buffer, for the rest species, by the Galbraith’s buffer. Results: The genome sizes of K. interior, K. heteroclita, K. longipedunculata, and K. coccinea were 7.36, 7.12, 7.01, and 5.15 pg/1C, respectively. Genome size of K. interior had no significant variation with those of K. heteroclita and K. longipedunculata (P = 0.296), which were larger than that of K. coccinea. Conclusion: Genome size did not distinguish K. interior, K. heteroclita and K. longipedunculata, but did distinguish from K. coccinea, which lays the foundation for future studies on genetic mechanism of the geoherb formation.

A novel technology to reduce astringency of tea polyphenols extract and its mechanism

Jin-yan Wan 1, Yu-lu Zhang 1, Yan Xiang, Nan Li, Ding-kun Zhang — 2021/8/2 9:37:08

Objective: Tea polyphenols are natural extracts used widely throughout the world. However, the severe astringency of tea polyphenols has reduced patient compliance. Based on the analysis of the formation mechanism of astringency, this paper hopes to propose a new method to control the astringency of tea polyphenols and improve patient compliance without changing its effect. Methods: Artificial saliva was used to prepare the tea polyphenols solution with different pH, using β-casein to imitate salivary protein, and preparing 1.2 mg/mL β-casein solution. A fluorescence quenching test was used to study the interaction between tea polyphenols and β-casein, combined with the stability test results of the compound, we can choose the pH with weak binding but good stability as the best pH for masking astringency. The taste-masking tablets were prepared under the best pH conditions, and the Xinnaojian Original Tablets were prepared according to the conventional preparation method. The disintegration time limit and solubility were tested respectively. The astringency of Xinnaojian original tablets and taste-masking tablets was evaluated by visual analogue scale (VAS). Results: The result of the fluorescence quenching test prompted that the combination force was the weakest when the pH was 4.9. Further synchronous fluorescence analysis showed that an increase in pH resulted in a decrease of the binding sites between tea polyphenols and β-casein, and this decrease was closely related to changes in tryptophan residues in β-casein. Both original and taste-masking Xinnaojian Tablets were prepared. Volunteers’ VAS scores illustrated that the astringency improved significantly with the masking tablets (P < 0.05). Conclusion: This pH-adjusting masking treatment had little effect on the recovery of polyphenols from the tablets or the dissolution of the tablets. This study provides a novel and feasible astringency masking technology for tea polyphenols and its preparation.

A strategy for rapid discovery of traceable chemical markers in herbal prod-ucts using MZmine 2 data processing toolbox: A case of Jing Liqueur

2021/8/2 9:37:09

Objective: The quality evaluation of herbal products remains a big challenge. Traceable markers are the core concept of the authentication of herbal products. However, the discovery of traceable markers is labor-intensive and time-consuming. The aim of this study is to develop a convenient approach to rapidly screen the traceable markers for herbal product authentication. Methods: Commercial Jing Liqueur and its 22 species of herbal ingredients were analyzed using HPLC-QTOF-MS and GC-MS to characterize nonvolatile and volatile chemicals. The acquired data were imported into MZmine 2 software for mass detection, chromatogram building, deconvolution and alignment. The aligned data were exported into a csv file and then traceable markers were selected using the built-in filter function in Excel. Finally, the traceable markers were identified by searching against online databases or publications, some of which were confirmed by reference standards. Results: A total of 288 chemical features transferred from herbal materials to Jing Liqueur product were rapidly screened out. Among them, 52 markers detected by HPLC-QTOF-MS were annotated, while nine volatile markers detected by GC-MS were annotated. Moreover, 30 of these markers were confirmed by comparing with reference standards. A chemical fingerprint consisting of traceable markers was finally generated to ensure the authentication and quality consistency of Jing Liqueur. Conclusion: A strategy for rapid discovery of traceable markers in herbal products using MZmine 2 software was developed.

Identification and verification of effective components of Huanghuai for dysfunctional uterine bleeding based on network pharmacology and molecular docking

2021/6/10 15:43:18

Objective: The Huanghuai (HH), which is made from the dried roots of Scutellaria baicalensis (Huangqin in Chinese) and the dried flowers and buds of Sophora japonica (Huaihua in Chinese), is a traditional Chinese formula used to treat dysfunctional uterine bleeding (DUB) (Benglou in Chinese) and proven to treat hemostasis effectively in our previous study. Network pharmacology and molecule docking were performed to study the underlying mechanism of Huanghuai (HH), and pharmacodynamic experiments were conducted to verify its curative effect. Methods: TCMSP, UniProt, GeneCards, STRING, DAVID databases, and Cytoscape 3.7.2 were utilized for the construction of a compound-target-pathway network. Docking the potential effective components with potential targets. The HPLC analysis of the potential effective components was performed. In vivo, the hot plate test model was used to study the analgesic activity, the egg white was used to study the swollen reaction in the sole in mice, and the hemostasis effect was studied by the capillary method, tail-breaking method and abortion uterus test. Results: The results showed that six compounds (acacetin, beta-sitosterol, wogonin, baicalein, kaempferol and quercetin) and four potential targets (PTGS2, AKT1, TP53 and TNF) in the compound-target-pathway network were the potential material basis for HH to treat DUB. It can be seen that the binding energy of the acacetin, wogonin, baicalein, beta-sitosterol, kaempferol and quercetin in HH docked with the receptor proteins PTGS2, AKT1, TP53, and TNF were far less than -5.0 kJ/mol, which means the molecules have low conformational energy, stable structure and high binding activity. And the result of HPLC analysis showed that acacetin, wogonin, baicalein, kaempferol and quercetin were the potential effective components of the hemostasis mechanism of HH, beta-sitosterol was removed due to low content. In vivo testing of the potential effective components, it revealed that the group of potential effective components identified by HPLC could increase the pain threshold, inhibit the swelling hind paws of mice induced by egg white, reduce the bleeding time and clotting time, reduce uterine bleeding, decrease the uterine weight, increase the content of Ca and ET-1, and reduce the content of NO in uterine homogenate tissue, and decrease of E2 and P content in uterine serum in aborted rats, whose efficacy was equal to HH. Conclusion: The results indicated that HH and potential active ingredient groups obtained from network pharmacology can treat DUB and play a hemostatic effect. The results obtained by network pharmacology have certain reliability. This study provides new indications for further mechanism research of HH on DUB and the development of HH or its components as an alternative therapy for patients with DUB. At the same time, the application of network pharmacology strategy may provide a powerful tool for exploring the mechanism of traditional Chinese medicine and discovering new biologically active ingredients.

Network pharmacology unveils spleen-fortifying effect of Codonopsis Radix on different gastric diseases based on theory of "Same treatment for different diseases" in traditional Chinese medicine

Ru-pu He, Zheng Jin, Ru-yun Ma, Fang-di Hu, Jian-ye Dai — 2021/6/10 15:43:18

Objective: "Same treatment for different diseases" is a unique treatment strategy under the guidance of traditional Chinese medicine (TCM) theory. Codonopsis Radix (Codonopsis pilosula, Dangshen in Chinese) with spleen-fortifying effect was employed to understand the strategy of "Same treatment for different diseases", based on its common mechanism in the treatment of gastric diseases including gastric ulcer, gastritis and gastric cancer via network pharmacology research. Methods: Network pharmacology research methods were used to analyze the interaction network and potential mechanisms of Dangshen in treating gastric ulcer, gastritis and gastric cancer. The active components and their target proteins of Dangshen were integrated from TCMSP, BATMAN-TCM databases. The targets of gastric ulcer, gastritis and gastric cancer were collected through GeneCards, PubMed, TDD and DisGeNET Database. Through screening, the key components and the key targets of Dangshen in treating gastric ulcer, gastritis and gastric cancer were obtained. After KEGG pathway analysis and GO analysis, the important pathways and biological processes were analyzed. Results: Through data and literature mining, the common and specific pharmaceutical effects and mechanism of Dangshen were summarized in these three gastric lesions. It was shown that Dangshen mainly acted on gastric ulcer, gastritis and gastric cancer through the overall regulation of the PI3K-AKT signaling pathway. With the development of the disease, it will gradually increase the control of inflammation through TNF, NF-κB and other inflammation-related signaling pathways to reduce inflammatory damage. For tumorigenesis, it pays more attention to inhibiting the ErbB signaling pathways to reduce the proliferation and migration of tumor cells. In addition, Dangshen's regulation of HIF-1 signaling pathway may also be beneficial for the treatment of gastric ulcer, gastritis and gastric cancer. Conclusion: Dangshen achieves spleen-fortifying effect on gastric diseases including gastric ulcer, gastritis and gastric cancer through multiple targets in multiple pathways, especially PI3K-AKT pathway and HIF-1 pathway. It could provide a scientific basis for understanding the strategy of "Same treatment for different diseases" in traditional Chinese medicine.

Hypoglycemic effects of Tibetan medicine Huidouba in STZ-induced diabetic mice and db/db mice

2021/6/10 15:43:18

Objective: Huidouba (HDB) is a Chinese folk medicine used to treat diabetes in Sichuan Province, China. Therefore, we investigated the anti-diabetic effects of HDB and its underlying mechanisms. We hypothesized that HDB treatment could enhance glucose tolerance and insulin sensitivity, and thus prevent a hyperglycemia state. Methods: To test the hypothesis, streptozotocin (STZ)-induced diabetic mice and db/db mice, widely used models of hyperglycemia and insulin-resistant diabetes, were either treated with HDB, metformin, or acarbose. Blood glucose, oral glucose tolerance test, insulin tolerance test, pancreatic histopathology and serum biochemistry were detected to assess the hypoglycemic effect of HDB. Results: HDB treatments were found to show the effect in reducing glucose levels. HDB also resulted in a significant reduction in body weight and food intake in the STZ-induced diabetic mouse model. Furthermore, it significantly improved glucose and insulin tolerance in the two diabetic mouse models. Importantly, insulin, glucagon, pancreatic polypeptide, and somatostatin immunohistochemistry revealed that HDB treatment improved the function and the location of the cells in the islets compared with the other two treatments. HDB treatment resulted in significant restoration of islet function. Our results illustrated the underlying mechanism of HDB in the progression of diabetes, and HDB can be an effective agent for the treatment of diabetes. Conclusion: The results of this study suggest that HDB can reduce blood glucose levels in STZ-induced hyperglycemic mice and db/db mice.

A six-herb Chinese medicine composition ointment as a promising candidate for treatment of hypertrophic scars

2021/6/10 15:43:18

Objective: To study the anti-hypertrophic scar effect of the six-herb Chinese medicine composition (SCMC) ointment on the rabbit ear hypertrophic scar models. Methods: The optimal formulation of SCMC ointment matrix was screened by the orthogonal designs and a series of evaluation tests. The SCMC ointment was prepared through emulsifying method. The rabbit ear hypertrophic scar models were established and used to investigate the anti-hypertrophic scar effect of SCMC ointment. Results: Our results demonstrated that all the quality control indications of the SCMC ointment met the requirements. Anti-hypertrophic scar activity results showed that all the rabbit ear scar tissues appeared different degrees of shrink and fading, and took an unobvious but palpable shift from hard to soft texture with the low, middle and high concentration SCMC ointments treatments in vivo. Additionally, on 21st day the scar area and thickness in different concentrations of SCMC ointment groups were significantly reduced than control group, in a concentration-dependent manner. The immunohistochemical results also indicated that the SCMC ointment had good anti-hypertrophic scar properties and could inhibit hypertrophic scar formation. Conclusion: The SCMC ointment could improve the blood circulation condition of hypertrophic scar tissues. Our research has demonstrated the Chinese medicine composition ointment with good anti-hypertrophic scar properties that could be used to treat hypertrophic scars. Meanwhile, it provides a theoretical basis for further clinical application.

Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells

Yu Xie, Ling Zhang, Yuan-yuan Li, Dian He, Li-fang Zheng — 2021/6/10 15:43:18

Objective: Chrysophanol (Chry) displays potent anticancer activity in human cancer cells and animal models, but the cellular targets of Chry have not been fully defined. Herein, we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity. Methods: Human liver cancer cell line HepG2 was incubated. The cytotoxicity was evaluated by MTT assay. Mitochondria localization was evaluated by a confocal microscopy. Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer. The levels of ATP, mitochondrial superoxide anions, and GSH/GSSG were determined according to the assay kits. The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining, respectively. The expression of cyclophilin D (CyPD) was determined by immunoblot method, and the interaction between CyPD and Chry was analyzed by molecule docking procedure. Results: Chry itself mainly localized in mitochondria to cause mitochondrial dysfunction and cell death in HepG2 cells. As regard to the mechanism, cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore (mPTP) moderately suppressed cell death, indicating mPTP involved in the process of cell death. Further, Chry enhanced the protein expression of Cyclophilin D (CyPD) which is a molecular componentry and a modulator of mPTP, while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD. Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with -11.94 kal/mol of the binding affinity value. Besides, the mtDNA-deficient HepG2-ρ0 cells were much resistant to Chry-induced cell death, indicating mtDNA at least partly participated in cell death. A combination of Chry and VP-16 produced the synergism effect toward cell viability and ΔΨm, while Chry combined with Cis-Pt elicited the antagonism effect. Conclusion: Taken together, enrichment in mitochondria and actions on mPTP, CyPD and mtDNA provides an insight into the anticancer mechanism of Chry. The combination therapy for Chry with clinical drugs may deserve to further explore.

Co-cultured adventitious roots of Echinacea pallida and Echinacea purpurea inhibit lipopolysaccharide-induced inflammation via MAPK pathway in mouse peritoneal macrophages

Ming-zhi Fan, Xiao-han Wu, Xue-feng Li, Xuan-chun Piao, Jun Jiang, Mei-lan Lian — 2021/6/10 15:43:19

Objective: In order to elucidate the biological activity of the co-cultured adventitious roots (ARs) of Echinacea pallida and Echinacea purpurea and provide theoretical basis for its application, and the anti-inflammatory activities and potential mechanisms of co-cultured ARs were studied. Methods: The experimental materials were obtained by bioreactor co-culture technology and used in the activity research. In this study, mouse macrophages induced by lipopolysaccharide (LPS) were used as in vitro model. Different concentrations of AR extract (50?400 g/mL) were used to treat cells. The expression of pro-inflammatory cytokines was determined using enzyme linked immunosorbent assay. The inducible nitric oxide synthase and cyclooxygenase-2 expression, mitogen-activated protein kinase (MAPK) phosphorylation, and the inhibitor of nuclear factor-kappa B-α levels were determined by the Western blot analysis. Results: In the co-cultured ARs, total flavonoids and total caffeic acid were determined, and the contents of both bioactive compounds were significantly higher than those ARs from the single-species culture. Compared with the control group, the large amount of pro-inflammatory mediators was released after LPS stimulation. However, in the extract groups with different concentrations (25, 50, and 100 g/mL), the production of these pro-inflammatory mediators was inhibited in a dose-dependent manner. Furthermore, the levels of phosphorylation of MAPK proteins, including p-p38, p-c-Jun N-terminal kinase, and p-extracellular regulated protein kinases were significantly (P < 0.05) decreased in the extract groups, revealing that the AR extract probably involved in regulating the MAPK signaling pathway. Conclusion: Collectively, our findings suggested that the co-cultured ARs of E. pallida and E. purpurea can inhibit production of pro-inflammatory mediators in mouse peritoneal macrophages and possess the anti-inflammatory effect by regulating MAPK signaling pathways.

Traditional Chinese herbal medicine Astragalus Radix and its effects on intestinal absorption of aconite alkaloids in rats

2021/6/10 15:43:19

Objective Astragalus Radix (AR, Huangqi in Chinese) has been widely used as a qi (energy) restoring herb that is thought to act through reinvigorating the spleen and lung. Aconite is used to rebalance the body temperature during illness and played an irreplaceable role in disease control since ancient times, but it is limited by its strong neuro and cardiotoxicity. Since the Song Dynasty (1227), the two herbs have been commonly used as herbal pairs including in the famous Qifu Decotion, from the “Wei's Family Prescription”. However, many ancient texts also record that they are not compatible using together, suggesting they can have negative outcomes when mixed. This study investigated whether Astragali Radix had either positive or negative effects on absorption of six different active alkaloids derived from aconite. Methods Single intestinal perfusion model was used to study the effects of Astragali Radix on aconite alkaloids absorption. Response of ABC transporters and distribution of three tight junction proteins on the surface of intestinal enothelium were assessed by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), Western blot and immunofluorescence microscopy, respectively. Results The results showed that aconite alkaloids absorption could be inhibited, and different concentrations of Astragali Radix considerably increased the expression levels of the ABC transporters and tight junction proteins with Astragali Radix treatment. Conclusion These results suggest that Astragali Radix can block absorption of aconite alkaloids through the upregulation expression of ATP-binding cassette transporters (ABC transporters) and tight junction proteins. It demonstrates that co-administration of Astragali Radix with other drugs might change the absorption profile of the second drug which is important to know in clinic therapy.

Evaluation of anti-inflammatory and anti-arthritic property of ethanolic extract of Clitoria ternatea

K. P. Swathia, Saravanan Jayaramb, Deepa Sugumarb, Emdormi Rymbaib — 2021/6/10 15:43:19

Objective: Clitoria ternatea is a well-known bioactive plant used to treat several inflammatory ailments in Ayurvedic system of medicine in India. The present investigation aimed to determine the anti-inflammatory and anti-arthritic activity of ethanolic extract of Clitoria ternatea roots (EECT) in animal models. Methods: The anti-inflammatory activity of the EECT was evaluated by carrageenan and histamine-induced paw edema. Results: EECT showed a significant reduction in mean paw edema volume in both carrageenan and histamine-induced inflammation. The efficacy of EECT in rheumatoid arthritis was tested against Freund’s complete adjuvant (CFA) induced arthritic models in Wistar rats. The anti-arthritic effect of EECT was determined by systematic scoring of arthritis symptoms and measuring paw edema. A considerable decrease in paw diameter was observed in the EECT (200 and 400 mg/kg) and diclofenac (10 mg/kg) treated groups after day 7. Diclofenac (10 mg/kg) and EECT (400 mg/kg) showed a significant reduction in paw diameter from day 14 compared with CFA control (P < 0.001). The anti-arthritic activity was also confirmed from the altered biochemical, haematological (Hb, RBC and WBC) and anti-oxidant parameters (SOD, MDA, CAT, and GSH). EECT (400 and 200 mg/kg) also showed a marked inhibition of joint destruction. Conclusion: This study provides a pharmacological rationale for the traditional use of C. ternatea against inflammation and rheumatoid arthritis in India.

Insights for integrative medicinal potentials of Ethiopian Kale (Brassica carinata): Investigation of antibacterial, antioxidant potential and phytocompounds composition of its leaves

2021/6/10 15:43:19

Objective: To compare the phytochemical and antimicrobial activity of Ethiopian Kale leaves infusions, investigate the antioxidant activity and profile the major phytochemicals existing in the better solvent system. Methods: Ethiopian Kale leaves were collected from Addis Ababa, Ethiopia, and extracted using different solvents. The qualitative phytochemical analysis, antibacterial assays, and Fourier Transform Infrared (FTIR) analysis are executed for all extracts. Antioxidant assay and Gas Chromatography-Mass Spectrometry (GC-MS) analysis are carried out for the solvent system, which showed better activity in preliminary studies. Results: The qualitative phytochemical analysis exposed the presence of different classes of phytoconstituents in most of the tested extracts. The broad spectrum of antibacterial activity (7–15 mm) was noted against the tested bacterial species. The functional groups of the extracts are reported by FTIR analysis. The antioxidant ability of ethanol extract was found to be (62.92 ± 0.34)% for 2,2-diphenyl-1-picrylhydrazyl (DPPH*) assay and (71.12 ± 0.41)% for 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assay. More than 17 major phytocompounds in ethanol extract was profiled by GC-MS analysis. Conclusion: The ethanol extract of Ethiopian Kale leaves contain a good source of phytochemicals and it can be a significant source for various functional applications.

Tetrahydroxystilbene glucoside protects against sodium azide-induced mitochondrial dysfunction in human neuroblastoma cells

Ru-yi Zhanga, Xu Zhangb, Lan Zhanga, Yan-chuan Wub, Xue-jing Sunc, Lin Lia — 2021/6/10 15:43:19

Objective: Mitochondrial dysfunction is evident in the early stage of Alzheimer’s disease (AD). Therefore development of drugs that protect mitochondrial function is a promising strategy for AD. The present work was to investigate the effects of 2, 3, 5, 4’-Tetrahydroxystilbene-2-O-β-d-glucosides (TSG) on a mitochondrial dysfunction cell model induced by sodium azide and elucidate the underlying mechanisms. Methods: Mitochondrial membrane potential (MMP) was detected by a fluorescence method. Cellular adenosine triphosphate (ATP) level was measured using a firefly luciferase-based kit. Reactive oxygen species (ROS) was detected using dichlorofluorescin diacetate (DCFH-DA). The expression levels of Bcl-2 and Bax were measured by Western blotting assay. Flow cytometry was utilized to measure apoptosis. Results: Pretreatment of TSG (25?200 mol/L) for 24 h significantly elevated MMP and ATP content, reduced ROS level and Bax/Bcl-2 ratio, and inhibited apoptosis in SH-SY5Y cells exposed to sodium azide. Conclusion: These results suggest that TSG protects SH-SY5Y cells against sodium azide-induced mitochondrial dysfunction and apoptosis. These findings are helpful to understand the protective effect of TSG on mitochondria, which are involved in the early stage of AD.

Collagen derived species-specific peptides for distinguishing donkey-hide gelatin (Asini Corii Colla)

2021/6/10 15:43:19

Objective: As an important food therapy product with traditional Chinese medicine (TCM) applications, donkey-hide gelatin (Asini Corii Colla, ACC) has been used for thousands of years. However, till now no effective strategy had been proposed to distinguish ACC from other animal hide gelatins, especially closely related horse- and mule-hide gelatins, which was an embarrassment of ACC quality control. Methods: Combined mass spectrometry and bioinformatic methods have been applied to identify and verify two ACC-specific peptides (Pep-1 and Pep-2) capable of distinguishing ACC from other closely related animal gelatins with high selectivity. Results: It confirmed that these two peptides could be not only used for distinguishing ACC from highly homologous horse-hide and mule-hide gelatins as well as other animal hide gelatins. Conclusion: The present study provides a simple method for species-specific peptides discovery, which can be used for assessing the quality of animal gelatin products, and ensure they are authenticable and traceable.

Suitable soil moisture contents for water use efficiency and saponins accumulation in Panax notoginseng

2021/6/10 15:43:19

Objective: The moisture content in the soil directly affects the yield and quality of Panax notoginseng, especially at the age of three years old. However, the suitable moisture for the growth of P. notoginseng is unknown. In this study, the effects of different soil moisture on the growth of P. notoginseng were studied. Methods: Four different water treatments (0.45 field capacity (FC), 0.60 FC, 0.70 FC, and 0.85 FC) were set up in Shilin County, Yunnan Province, China. The water consumption and daily dynamic of water consumption were determined daily (from April 21 to October 18, 2012), and the daily dynamic of water consumption under different weather conditions (sunny and rainy) was determined. The transpiration coefficient and water use efficiency were calculated through dry matter accumulation and total water consumption. Accumulation of saponins of roots of P. notoginseng were analyzed by HPLC after treated, and the soil moisture content suitable for the growth of P. notoginseng was estimated by regression fitting of the active ingredient accumulation and the soil moisture content. Results: The water consumption of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.89, 3.68, 3.37 and 2.73 kg/plant per day, respectively. The water consumption of P. notoginseng from June to August was greater than other months. The daily dynamic of water consumption on sunny days and sunny days after rain showed a “double peak” feature, and it showed a “single peak” feature on rainy days. The water uses efficiency (WUE) of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.51, 3.32, 4.59, 3.39 gDW/kg H2O, respectively. The increase of soil moisture content would reduce the WUE of P. notoginseng. With the increase of soil water content, the content of notoginsenoside R1 and ginsenoside Rg1 did not change significantly, while the content of ginsenoside Rb1 and Rd showed a decreasing trend. Conclusion: Soil moisture content significantly affected the water consumption of P. notoginseng, and when it was 56.4% of the maximum water holding capacity in the field, the sum of the four saponins of 100 strains of P. notoginseng was the highest.

Formulation optimization and characterization of transdermal film of curcumin by response surface methodology

Priyanka Kriplani1,2, Kumar Guarve1, Uttam Singh Baghel3 — 2021/6/10 15:43:19

Objective: India is referred as goldmine of herbal drugs but still lack of optimization of herbal drugs, which has kept us on the back foot. The rationale of the study is to prepare optimized transdermal drug delivery system of curcumin employing response surface methodology to study the collective effect of independent variables like concentration of ethyl cellulose, hydroxyl propyl methyl cellulose and dibutyl phthalate which significantly influenced characteristics like percentage elongation and in vitro drug release. Method: Twenty formulations containing varying concentrations of polymers and permeation enhancer were prepared using solvent casting technique. Result: The study revealed that the effect of dibutyl phthalate (DBP) concentration was the highest on percentage elongation (P < 0.0001), while hydroxy propyl methyl cellulose (HPMC) concentration exhibited pronounced effect on drug release (P < 0.0001) through dialysis membrane. Linear model fitted the best for curcumin release and elongation for all formulations. According to Derringer’s desirability prediction tool, the composition of optimized film was found to be 242.14% of HPMC, 109.61% of ethyl cellulose (EC), and 1.03% of DBP. Under these conditions, the optimized patch exhibited a predicted value of %elongation and in vitro drug release of 94.35% and 80.0306%, respectively, which was comparable to the actual values of percent elongation and in vitro drug release i.e. 95.02% and 81.03% respectively. FTIR and thermal studies were also performed which revealed no interaction or complexation between drug and excipients. The ex vivo study performed using rat skin showed that the cumulative drug release from the optimized patch showed flux of (30.68 ± 18) μg/cm2/h. Conclusion: It can be concluded that in future if proper optimization of herbal formulations is carried out, they can become the first choice for patients as compare to synthetic drugs.

Gene mining and efficient biosynthesis of a fungal peptidyl alkaloid

2021/1/18 13:59:00

Objective: Peptidyl alkaloids, a series of important natural products can be assembled by fungal non-ribosomal peptide synthetases (NRPSs). However, many of the NRPSs associated gene clusters are silent under laboratory conditions, and the traditional chemical separation yields are low. In this study, we aim to discovery and efficiently prepare fungal peptidyl alkaloids assembled by fungal NRPSs. Methods: Bioinformatics analysis of gene cluster containing NRPSs from the genome of Penicillium thymicola, and heterologous expression of the putative gene cluster in Aspergillus nidulans were performed. Isolation, structural identification, and biological evaluation of the product from heterologous expression were carried out. Results: The putative tri-modular NRPS AncA was heterologous-expressed in A. nidulans to give anacine (1) with high yield, which showed moderate and selective cytotoxic activity against A549 cell line. Conclusion: Heterologous expression in A. nidulans is an efficient strategy for mining fungal peptidyl alkaloids.

Neuroprotective effects of kukoamine A on 6-OHDA-induced Parkinson's model through apoptosis and iron accumulation inhibition

2021/1/18 13:59:00

Objective: Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in substantia nigra (SN). Our previous study demonstrated kukoamine A (KuA) to exhibit strong neuroprotective effects through antioxidative stress, and autophagy in MPTP/MPP+-induced PD models in vivo and in vitro. It is necessary to evaluate the efficacy of the anti-PD effects under various models. Methods: In the present study, total chemical synthesis was used to obtain KuA, which performed low content in Lycii Cortex. Then, 6-OHDA-induced PD model of PC12 cells was used to investigate the effects of KuA on PD. Results: Our results demonstrated that KuA ameliorated cell loss and mitochondrial membrane potential (MMP) loss, and inhibited Bax/Bcl-2 ratio increase that were induced by 6-OHDA. Iron accumulation in SN is thought to participate in neuronal death in PD, which subsequently resulted in oxidative stress and overexpression of α-synuclein caused by iron metabolism protein disorder. In our study, KuA could chelate cellular iron content and decrease iron influx. Moreover, KuA could upregulate the expression of ferroportin1 and Hephaestin, downregulate the expression of DMT1, TfR, and Ferritin to maintain cellular iron homeostasis avoiding neuronal death from cellular iron deposition. Moreover, KuA could decrease the expression of a-synuclein in cells. All the results indicated that KuA protected against neurotoxin-induced PD due to the apoptosis inhibition and iron homeostasis maintaining. Conlusion: KuA treatment might represent a neuroprotective treatment for PD.

Berberine exerts antidepressant-like effects via regulating miR-34a-synaptotagmin1/Bcl-2 axis

Li-tao Yi a, Ji-xiao Zhub, Shu-qi Donga, Min Chena, Cheng-fu Lic — 2021/1/18 13:59:00

Objective: Berberine, a cationic alkaloid first isolated in 1917, has been approved by the China Drug Administration for decades. Accumulating evidence demonstrated its antidepressant-like activities in vivo. Our previous study has shown that chronic stress leads to the upregulation of miR-34a in the hippocampus of mice. This study aims to evaluate the underlying miR-34a mediated mechanism of berberine in chronic stress-induced depression in mice. Methods: In the present study, mice were administered with berberine during chronic stress. Levels of miR-34a, dendritic density, mitochondrial morphology, and neurogenesis were assessed in the hippocampus. Subsequently, miR-34a agomir was used as a pharmacological intervention for the investigation of berberine. Results: The results showed that berberine reversed the decrease in sucrose preference and the increase in latency to feed without altering total food consumption. Furthermore, chronic stress-induced overexpression of miR-34a decreased synaptotagmin-1 and Bcl-2 levels, thereby impairing spinal morphology, mitochondria and neurogenesis. Berberine inhibited miR-34a expression, in turn restored synaptotagmin-1 and Bcl-2 levels, and thus improved spinal morphology, mitochondria and neurogenesis in the hippocampus. However, the improvements induced by berberine were totally blocked by the pretreatment of miR-34a agomir, which caused the elevation of miR-34a levels in the hippocampus. Conclusion: This finding demonstrated that miR-34a downregulation was involved in the antidepressant-like effects of berberine in mice exposed to chronic stress.

Cytotoxicity of oleanane type triterpene from leaf extract of Pterospermum acerifolium (in vitro) and theoretical investigation of inhibitory signaling pathway

Mohd Rehan, Shafiullah Sultanat — 2021/1/18 13:59:00

Objective: To evaluate the cytotoxic activity of taraxerol isolated from the leaves of Pterospermum acerifolium and its EtOH extract against human breast, colon, and lung cancer cell lines and docking studies. Methods: The structures of the isolated compounds were elucidated by several spectroscopic methods such as 1H-NMR, 13C-NMR, DEPT-135, COSY, HSQC, and HMBC. The extract and isolated compound were analyzed for cytotoxic activity on MDA-MB-231, BT-549, A-549, and SW-480 cancer cell lines by MTT assay. Molecular docking was performed on software such as Chem3D pro 12.0.2.1076, Discovery Studio Visualizer, Auto Dock Tools-1.5.6 and Auto dock vina. Results: The extract and isolated compound taraxerol both displayed excellent inhibitory activity (IC50: 80 μg/mL for extract and 160 g/mL for compound) on breast cancer cell (MDA-MB-231). The docking studies show a strong affinity with PI3K (-9.8 Kcal/mol) and mTOR (-10.0 Kcal/mol). Conclusion: The results confirm that the extract and compound exhibited strong cytotoxicity on the MDA-MB-231 cancer cell. So, the extract and the compound may be useful in human chemotherapies.

Induction of long-term potentiation at Schaffer collateral-CA1 synapses in mice hippocampus after IMPX977 administration

2021/1/18 13:59:00

Objective To investigate the effects of IMPX977 on long term potentiation (LTP) at Schaffer collateral-CA1 synapses in vitro and on methyl CpG binding protein 2 (Mecp2) expression in mice cortex and hippocampus. Methods Thirty-two C57BL/6 mice were randomly divided into four groups: control, olive oil (vehicle), IMPX977 low (5 mg/kg) and high (15 mg/kg) groups. Mice were administrated every other day orally for two weeks. Extracellular recording technique in vitro was used to record the effects of IMPX977 on Schaffer collateral-CA1 LTP pathway in acute mice hippocampal slices. The Mecp2 protein expression level was detected by Western blotting. Results Compared to the control group, vehicle did not alter the synaptic transmission in Schaffer collateral-CA1 synapses, however, IMPX977 at concentrations of 5 mg/kg and 15 mg/kg significantly enhanced fEPSP (field excitatory postsynaptic potential) slope in Schaffer collateral-CA1 pathway to (179.6 ± 17.8)% and (191.4 ± 21.4)%, individually 60 min after HFS, IMPX977 improved LTP induction significantly at Schaffer collateral-CA1 pathway at least. Also, IMPX977 significantly elevated MeCP2 protein level in cortex and hippocampus. Conclusion The effects of IMPX977 on synaptic transmission and Mecp2 protein expression provided convincing evidence that IMPX977 could be promising new drug candidates for Rett syndrome treatment.

Effect of green tea consumption in treatment of mild to moderate depression in Iranian patients living with HIV: A double-blind randomized clinical trial

2021/1/18 13:59:00

Objective: Depression affects people living with HIV (PLWH) compliance leading to poor control infection. Previous observational studies showed an anti-depression effect of green tea extract (GTE). The therapeutic effect of GTE on depression were investigated in PLWH receiving antiretroviral therapy (ART). Methods: Fifty PLWH on ART with diagnose of mild to moderate of depression, participated in a double-blind, placebo-controlled trial and underwent 12 weeks of treatment with either 400 mg GTE capsules or placebo twice daily. The Hamilton depression scale of patients was measured before, 6 weeks and 12 weeks after treatment in two groups. The primary outcome measure was performed to evaluate the efficacy of GTE in improving depressive symptoms. Results: The mean of Hamilton score showed a significant difference between the two groups after 12 weeks (P = 0.035). Repeated measures ANOVA test showed a significant effect for time × treatment interaction on the Hamilton mean score between the two groups (P = 0.000). Conclusion: It seems the use of GTE capsules in PLWH on ART is safe and could lead to greater and more rapid improvement in depressive symptoms than placebo. Thus it can be considered as an alternative therapy for mild to moderate depression. Further studies with higher sample size and longer follow-up and comparisons with other antidepressive drugs are warranted.

Comparative authentication of Semiliquidambar cathayensis and its substituted species via macroscopic and microscopic features

Dan Zhua, Xincheng Qub, Xuemei Suna, Shihuan Yana, Hongwei Guoa, Yaoli Lic — 2022/11/18 8:20:25

Objective: Ban Fenghe recorded in the Quality Standard of Yao Medicine of Guangxi Zhuang Autonomous Region (Volume 1) is derived from the dried stems and leaves of Semiliquidambar cathayensis. It is usually confused with medicinal herbs from Pterospermum heterophyllum and Dendropanax dentiger. However they are very different in chemical composition, and should not be used as the same drug. To ensure their safety and efficacy, a method based on macroscopic and microscopic characteristics was developed to distinguish them. Methods: A total of 14 batches of Ban Fenghe samples from three species were collected from different producing areas in China. The macroscopic characteristics were examined by observing external traits. The tissue structures of transverse sections of stems and leaves, the leaf epidermis, and the powder were observed microscopically. Results: The branchlets and leaf surfaces of S. cathayensis and P. heterophyllum were hairy, especially the lower leaf surfaces of P. heterophyllum were densely covered with hairs, but those of D. dentiger were hairless. The pericyclic fibers of S. cathayensis stems were intermittently distributed in a circular shape and accompanied by stone cells, whereas those of P. heterophyllum and D. dentiger were bundled without stone cells. So stone cells and hairs were present in S. cathayensis powder, stone cells were not found in P. heterophyllum and D. dentiger powder, and hairs were not present in D. dentiger powder. The distribution sites, sizes and types of secretory tissues of these three species were also different in transverse sections of stems and leaves. Stomata on the lower epidermis of S. cathayensis leaves were paracytic, whereas those of P. heterophyllum and D. dentiger were anomocytic. Conclusion: Ban Fenghe drugs derived from S. cathayensis could readily be distinguished from those of P. heterophyllum and D. dentiger by macroscopic and microscopic features.

Specification and grade of Saposhnikoviae Radix (Saposhnikovia divaricata)

2022/11/18 8:20:25

Objective Saposhnikoviae Radix (Fangfeng in Chinese), the roots of Saposhnikovia divaricata, lacks commodity specification and grade standardization in the current market. This study investigated the existing specifications and grades of Saposhnikoviae Radix to provide a standardized scientific reference for its market use. Methods Based on a textual research of Chinese herbal medicine from the Han Dynasty to the present, medicinal materials of different specifications and grades obtained from Saposhnikoviae Radix in the main producing areas of China were collected and the markets for these materials were investigated. Field investigations were performed in the major producing areas such as Northeast China, Hebei Province, and Inner Mongolia. Four major Chinese herbal medicine markets in China were investigated. Sensory indices were used to categorize the two specifications (wild and cultivated) according to the shape, color, texture, and cross-section. High-performance liquid chromatography was performed to determine the active components. Vernier calipers and measuring tape were used to measure the diameter and length, respectively, of 41 samples. Using Excel and the R Language software, cluster analysis and descriptive statistical analysis were performed to assist in the application of new specifications and grades based on physical characteristics, pharmacological activity, and chemical composition. Results The two specifications (wild and cultivated) of Saposhnikoviae Radix were divided into three grades each based on the length and diameter. Prim-O-glucosylcimifugin, 5-O-methylvisamminoside, and the length of Saposhnikoviae Radix can be used as a basis for classifying the commodity specifications and grades. The specifications and grade standards of Saposhnikoviae Radix were established based on the following eight aspects: shape, surface characteristics, texture, cross section, taste, prim-O-glucosylcimifugin content, 5-O-methylvisamminoside content and length. Conclusion The formulation of this standard stipulates the commodity specification level of Saposhnikoviae Radix. It is also suitable for the evaluation of commodity specifications in the process of production, circulation and use of Saposhnikoviae Radix.

Taxifolin attenuates inflammation via suppressing MAPK signal pathway in vitro and in silico analysis

2022/11/18 8:20:25

Objective Taxifolin is a natural flavonoid compound that can be isolated from onions, grapes, oranges and grapefruit. It also acts as a medicine food homology with extraordinary antioxidant and anti-inflammatory activity. This study aims to explain the protective effects and potential mechanisms of taxifolin against inflammatory reaction. Methods Levels of interleukin (IL)-6, IL-1β and intracellular reactive oxygen species (ROS)were assessed in different time after the treatment of taxifolin in RAW264.7 cells induced by lipopolysaccharide (LPS). Subsequently, the mRNA and protein levels of inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α and the phosphorylation expression levels of the MAPK signal pathway were also evaluated. A silico analysis was used to explain the binding situation for the investigation of taxifolin and MAPK signal pathway. And then MAPK inhibitors were used to reveal the expression level of iNOS, VEGF, COX-2 and TNF-α in RAW264.7 cells. Results It was demonstrated that cell inflammatory damage induced by LPS was significantly alleviated after the treatment of taxifolin. Then, the mRNA and protein levels of iNOS, VEGF, COX-2 and TNF-α were reduced and the phosphorylation expression levels of the MAPK signal pathway were down-regulated remarkably as well. In silico analysis, taxifolin could form a relatively stable combination with MAPK signal pathway. MAPK inhibitors showed increasing or decreasing effect in the mRNA levels of iNOS, VEGF, COX-2 and TNF-α, which suggesting that taxifolin down-regulated iNOS, VEGF, COX-2 and TNF-α expressions were not entirely through the MAPK pathway. Conclusion This finding demonstrated that taxifolin improved the inflammatory responses that partly involved in the phosphorylation expression level of MAPK signal pathway in RAW264.7 cells exposed to acute stress.

Integrating systematic pharmacology-based strategy and experimental validation to explore mechanism of Tripterygium glycoside on cholangiocyte-related liver injury

Yajing Li, Sen Lia, Xiaoyong Xuea, Ting Wangb, Xiaojiaoyang Lia — 2022/11/18 8:20:25

Objective: Tripterygium glycoside(TG) is widely used in clinical practice for its multiple bioactivities including anti-inflammatory and immunosuppressive effects. However, emerging studies have frequently reported TG-induced adverse reactions to multiple organs, especially liver. Here, this studyaimed to investigate the mechanism of liver damage induced by TG and explore representative components to reflect TG hepatotoxicity. Methods: Network pharmacology was used to determine the potential targets of bile duct injury caused by TG. Next, the hepatotoxic effects of TG, triptolide (TP) and celastrol (CEL) were investigated and compared in vivo and in vitro. Liver function was determined by measuring serum transaminase and histopathology staining. The cell proliferation and apoptosis were determined by cell viability assay, scratch assay and flow cytometry. The expression of gene of interest was determined by qPCR and Western blot. Results: Based on the network pharmacological analysis of 12 bioactive ingredients found in TG, a total of 35 targets and 15 pathways related to bile duct injury were obtained. Both TG and TP resulted in cholangiocyte damage and liver injury, as illustrated by increased levels of serum transaminase and oxidative stress, stimulated portal edema and lymphocytic infiltration and decreased expression of cholangiocyte marker, cytoskeletal 19. In addition, TG and TP inhibited cell proliferation and migration, arrested cell cycle and promoted Caspase-dependent apoptosis of cholangiocytes via suppressing the phosphorylation of extracellular regulated protein kinases 1/2 (ERK1/2) and protein kinase B (AKT). While, CEL at equivalent dosage had no obvious hepatotoxicity. Conclusion: We revealed that TG-stimulated liver injury was specifically characterized by cholangiocyte damage and TP might be the decisive ingredient to reflect TG hepatotoxicity. Our results not only provide novel insights into the mechanism underlying the hepatotoxicity effects of TG but also offer reference for clinical rational use of TG.

Lithocarpus polystachyus (Sweet Tea) water extract promotes human hepatocytes HL7702 proliferation through activation of HGF/AKT/ERK signaling pathway

Ming Leia,b, Nana Chena,b, Yingshu Xua, Qihai Gongb, Jianmei Gaoa,b — 2022/11/18 8:20:25

Objective: Sweet Tea (ST), derived from the leaves of Lithocarpus polystachyus, is a Chinese folk medicine with wide pharmacological activities. However, the promotive effects of ST water extract on hepatocytes proliferation and its underlying mechanism remains still unknown. In the present study, the beneficial effects of ST water extract on human hepatocytes and its possible mechanism were investigated. Methods: MTT assay was used to detect the safety range of ST; HL7702 cells were divided into four groups: control group, ST low- (50 μg/mL), medium- (200 μg/mL) and high-concentration (800 μg/mL) groups; BrdU ELISA and EDU staining were used to observe DNA content and cell proliferation; Moreover, flow cytometry was applied to analyze the distribution of cell cycle. Furthermore, the expression of cyclin D1, CDK4, HGF/c-Met, Akt, Erk1/2 were detected by Western blot. Results: It was found that ST water extract concentration-dependent promoted human hepatocytes HL7702 cell proliferation within 72 h through accumulating the cells in S phase and G2/M phase. Furthermore, ST water extract up-regulated expression of Cyclin D1 and CDK4 proteins. Moreover, ST water extract not only increased HGF expression and phosphorylation of c-Met level, but also activated the phosphorylation levels of AKT, ERK1/2. Interestingly, both of AKT inhibitor A6730 and ERK1/2 inhibitor U0126 reversed the promotive effects of ST water extract, which further confirmed that activation of AKT and ERK1/2 were involved. Conclusion: The findings reveal that ST water extract promoted HL7702 cells proliferation through the stimulation of cell cycle mediated by activating the AKT- and ERK1/2-related pathway.

Reduning Injection prevents carrageenan-induced inflammation in rats by serum and urine metabolomics analysis

2022/11/18 8:20:25

Objective: To elucidate the anti-inflammatory mechanism of Reduning Injection (RDN) by analyzing the potential biomarkers and metabolic pathways of the carrageenan-induced inflammatory model from the overall metabolic level. Methods: Rat inflammatory model was established by carrageenan. UPLC-Q-TOF/MS was used to detect and analyze changes of endogenous metabolites in the serum and urine of carrageenan-induced inflammatory rats. Combined with multivariate analysis and databases analysis, inflammatory-related potential biomarkers were screened and identified to analyze possible metabolic pathways. The reliability and biological significance of these biomarkers was verified by metabolic network analysis and correlation analysis with pharmacodynamic indicators. Results: A total of 16 potential biomarkers were screened and identified by multivariate analysis and metabolite databases, among which 13 species could be adjusted by RDN. The metabolism pathway analysis revealed that histidine metabolism, sphingolipid metabolism, and tyrosine metabolism were greatly disturbed. Their biomarkers involved urocanic acid, sphingosine, and norepinephrine, all of which showed a callback trend after RDN treatment. The three biomarkers had a certain correlation with some known inflammatory-related small molecules (histamine, arachidonic acid, Leukotriene B4, and PGE2) and pharmacodynamic indicators (IL-6, IL-1β, PGE2 and TNF-α), which indicated that the selected biomarkers had certain reliability and biological significance. Conclusion: RDN has a good regulation of the metabolic disorder of endogenous components in carrageenan-induced inflammatory rats. And its anti-inflammatory mechanism is mainly related to the regulation of amino acid and lipid metabolism. This research method is conducive to the interpretation of the overall pharmacological mechanism of Chinese medicine.

Variation of photosynthesis, secondary metabolites and antioxidant activities in third generation of spaceflight-induced Salvia miltiorrhiza

Liang Penga,c, Mei Rud, Zongsuo Liangb,c — 2022/11/18 8:20:25

Objective: Spaceflight has long been perceived as an effective way to improve the quantity and quality of plants with wide applications. In order to obtain stable and inheritable descendants of spaceflight-induced Salvia miltiorrhiza lines, we investigated and analyzed four lines m16, m50, m51, m57 (three individuals of each line) and the ground control (three individuals) of the third generation of spaceflight-induced S. miltiorrhiza from primary/secondary metabolism and antioxidative abilities. Methods: A portable photosynthesis system (Li-6400) with red/blue LED light source was used to perform the photosynthetic characteristics to evaluate their primary productivity. The secondary metabolites (phenolic acids, tanshinones, total phenolics and flavonoids) and antioxidant activity of roots were analyzed to assess their quality. Results: Compared with control, line m16 presented weak photosynthetic ability, but high apparent quantum yield (AQY), higher contents of secondary metabolites, and stronger antioxidative abilities. Line m57 had a strong gas exchange ability, relatively higher secondary metabolites contents, and ascending antioxidative abilities. Lines m50 and m51 were in the middle level of lines m16 and m57. The principal component analysis for all the original data revealed three components including a root-related index, a leaf-related index, and a CO2 response parameter could be used to distinguish spaceflight-induced S. miltiorrhiza lines. Conclusion: Line m57 could be an appropriate material for the investigation of targeted breeding towards high production, and line m16 could be used to identify essential genes and unravel sophisticated pathways underlying the secondary metabolisms.

Metabolic regulation mechanism of Aconiti Radix Cocta extract in rats based on 1H-NMR metabonomics

2022/11/18 8:20:25

Objective To establish a metabonomics research technique based on the combination of 1H-NMR and multivariate statistical analysis, so as to explore the metabolic regulation mechanism of Aconiti Radix Cocta extract (ARCE) in rat tissues and serum. Methods SD rats were randomly divided into blank group, female group and male group. The 1H-NMR technique was used to collect the information of rat tissues and serum samples in each group. The principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and other methods were used for data pattern recognition, so as to screen out potential differential metabolites and metabolic pathways, and then network analysis and KEGG database were used to analyze the relationship between metabolites, metabolic pathways and diseases. Results The external features and 1H-NMR analysis showed that the sex of rats had no obvious effect on the drug action. A total of 15 potential differential metabolites and six metabolic pathways were screened out through data pattern recognition. Through network analysis and KEGG pathway analysis, three target diseases closely related to differential metabolites were found, and the metabolic pathway related to lung cancer was the central carbon metabolism of cancer. Conclusion This study shows that Aconiti Radix Cocta (ARC) may regulate the energy metabolism of the body by influencing arginine synthesis, so as to play the roles of anti-inflammation, analgesia, anti-tumor and immune regulation.

Identification of Q-Markers from Hedan Tablet by employing "spider-web" mode and taking compounds hepatotoxicity into account

2022/11/18 0:00:00

Objective: As a classic prescription in traditional Chinese medicine, Xuefu Zhuyu Decoction (XFZYD) has been widely used in the clinical treatment of cardiovascular and cerebrovascular diseases. In order to unveil the potentially effective compounds, a rapid ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) method was established to identify prototype components and their metabolites from XFZYD in rats' serum. Methods: The serum from rats after intragastric administration of XFZYD aqueous extract was analyzed by UPLC-Q-TOF/MS method. The prototype components and their metabolites were identified by comparison with the reference standards and tentatively characterized by comprehensively analyzing the retention time, MS data, characteristic MS fragmentation pattern and retrieving literatures. Results: A total of 175 components (24 prototype components and 151 metabolites) were identified and tentatively characterized. The metabolic pathways of prototype components in vivo were also summarized, including glucuronidation, hydrolyzation, sulfation, demethylation, and hydroxylation, and so on. Conclusion: In this study, a UPLC-Q-TOF/MS technique was developed to analyze prototype components and their metabolites from XFZYD in serum, which would provide the evidence for further studying the effective compounds of XFZYD.

Trace heavy metals and harmful elements in roots and rhizomes of herbs: Screening level analysis and health risk assessment

Xiaoli Wu, Peiling Wu, Mengying Gu, Jian Xue — 2022/11/18 8:20:25

Objective: Heavy metal and harmful element contamination are frequently reported in Chinese herbal medicines (CHMs), and roots and rhizomes parts showed a higher content than other parts. To investigate the residue level and assess the potential human health risk of heavy metals and harmful elements in roots and rhizomes, 720 batches of the sample representing 20 species of herbs from different sources were collected. Methods: The content of Pb, Cd, As, Hg, and Cu in the digests was determined using ICP-MS. The chronic hazard index estimate based on non-cancer hazard quotient (HQ) was applied for potential health risk assessment of Pb, Cd, As, Hg, and Cu via consumption of CHMs. Results: Compared with the Chinese limit standard (Chinese Pharmacopoeia Commission, 2020 edition) of Pb, Cd, As, Hg, and Cu in Ginseng Radix et Rhizoma, the exceedance percentage of Pb in total samples was 14.1%, which were generally far higher than Cd, As, Hg, and Cu. Health risk assessment results based on hazard quotient calculating showed that total HQ of Cu, Pb, As, Cd, and Hg in Pulsatillae Radix and Clematidis Radix et Rhizoma exceeded 1, with the value of 1.543 and 1.235. Besides, Arsenic had the highest HQ value (0.957) in Pulsatillae Radix. Conclusion: Consuming raw materials of Pulsatillae Radix and Clematidis Radix et Rhizoma may pose a potential risk and Arsenic residues in Pulsatillae Radix deserved special attention.

Comparative study on brain pharmacokinetics of Buyang Huanwu Decoction in normal and cerebral ischemia rats using brain microdialysis combined with LC-MS/MS

Wanyi Liu, Keqi Zeng, Xin Zhou, Yingfeng Zhang, Cong Nie — 2022/11/18 8:20:25

Objective: To conduct a comparative study on the brain pharmacokinetics of seven ingredients (i. e. senkyunolide A, ferulic acid, formononetin, calycosin, ononin, calycosin-O-β-D-glucopyranoside, and paeoniflorin), which were the compounds of Buyang Huanwu Decoction (BHD), in normal and cerebral ischemia rats administrated intragastrically with BHD. Methods: The samples of normal and permanent middle cerebral artery occlusion (pMCAO) rats were collected by using brain microdialysis technique. The concentrations of seven ingredients were determined by the HPLC-MS/MS method. After the BHD were administrated intragastrically to the rats for seven consecutive days, brain microdialysis probes were inserted into the hippocampus of rats, and then the brain microdialysates were collected at 20 min time intervals for 5 h. The separation of the seven ingredients and internal standard (IS) was carried out on an ACQUITY UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm) chromatographic column, using a mobile phase consisting of acetonitrile (containing 0.1% formic acid) and water (containing 0.1% formic acid) for gradient elution within 13 min. The ionization was conducted using an ESI source in positive ion mode. Multiple reaction monitoring mode was used for quantification of ingredients in BHD. Results: Linearity, accuracy, precision, matrix effect and stability of LC-MS/MS method were all satisfactory, successfully applied to compare the pharmacokinetics of the analytes between normal and model rats after intragastric administration of BHD. Compared with the normal group, the model group after the administration of the BHD showed that T1/2 of formononetin and ononin were longer, and except for calycosin-O-β-D-glucopyranoside (P < 0.01), there was no significant difference between the normal group and the model group. The Cmax of senkyunolide A and calycosin of model group were increased, while the Tmax of senkyunolide A was decreased, and except for the Tmax of PF, the differences between the two groups were statistically significant (P < 0.01). Conclusion: The LC-MS/MS method combined with microdialysis was successfully applied to the comparative study of brain pharmacokinetics of seven ingredients in BHD. After intragastric administration of BHD, there were differences in the pharmacokinetics of seven ingredients in the brain hippocampus between normal rats and model rats, probably related to the characteristics of the ingredients and the effects of cerebral ischemia on the absorption and distribution of the ingredients.

Camganoids A and B, two new sesquiterpenes with different carbon skeletons isolated from fruits of Cinnamomum migao

2022/11/18 8:20:25

Objective: To isolate and identify the undescribed compounds from the fruits of Cinnamomum migao and evaluate its nitric oxide inhibition potential. Methods: The chromatographic techniques of silica gel, sephadex, and HPLC were used for isolation and purification of the compounds, while HR-ESI-MS, 1D-NMR, 2D-NMR, ECD, and X-ray diffraction techniques were used to characterize and confirm the isolated compounds. Moreover, the anti-inflammatory activity of the isolated compounds was carried out to check inhibitory potential against the production of nitric oxide with RAW264.7 cells stimulated by LPS. Results: Camganoid A (1), a novel sesquiterpene possessing an unprecedented skeleton, and camganoid B (2), containing a unique eight-membered sesquiterpene moiety with a new carbon skeleton, were isolated and identified from the fruits of C. migao. The absolute configurations of 1 and 2 were confirmed by single crystal X-ray diffraction and electronic circular dichroism (ECD) calculations. Among these compounds, compound 1 exhibited potent inhibitory activity against the production of nitric oxide with IC50 value of 4.59 μmol/L in RAW264.7 cells stimulated by LPS. Conclusion: The isolation of two new skeletons from the fruits part of C. migao possessed unique skeletons which have not been reported before.

Effect of date palm pollen supplementation on female sexual function in non-menopausal women: A double blind randomized clinical trial

2022/11/18 8:20:25

Objective: Despite numerous experimental studies in the literature, there are few clinical trials regarding the effect of date palm pollen (DPP) supplementation on sexual function improvement. In the present study, we sought to evaluate the impact of DPP on female sexual function in Iranian non-menopausal women. Methods: Between October 2019 and December 2019, health centers in the city of Khalkhal, volunteers meeting the inclusion criteria were recruited in randomized clinical trials. Sixty-eight women were randomly stratified and assigned to one of the two study groups: placebo group (n = 35) and palm pollen group (n = 35), and received a starch or palm pollen capsule (300 mg per day), respectively, for 35 d. The Female Sexual Function Index (FSFI) instrument was used to assess female sexual function. Results: After DPP supplementation, the increase in desire, lubrication, and the overall score, was statistically significant compared to the placebo group (P = 0.002, P = 0.000, and P = 0.042; respectively); Whilst there was no significant differences in the remaining domains (arousal: P = 0.763; orgasm: P = 0.370; satisfaction: P =0.474; pain: P = 0.259). There was a statistically significant positive correlation between the coitus and preintervention levels of desire (r = 0.298, P = 0.038), arousal (r = 0.328, P = 0.021), lubrication (r = 0.361, P = 0.011), orgasm (r = 0.320, P = 0.025), satisfaction (r = 0.327, P = 0.022), and overall scores (r = 0.338, P = 0.018). Conclusion: This study suggests that DPP (300 mg supplementation for 35 d), given to non-menopausal women, could improve the lubrication and desire domains of FSFI.

Metabolomics investigation on antiobesity effects of Corydalis bungeana on high-fat high-sugar diet-induced obese rats

2022/9/1 15:43:52

Objective: Corydalis bungeana (CB) is a well-used medicinal herb in Mongolian folk medicine and has been traditionally applied?as?an antiobesity agent. However, the evidence-based pharmacological effects of CB and its specific metabolic alterations?in the obese model are not entirely understood. This study aimed to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite alterations associated with weight loss and lipid metabolism in obese rats. Methods: A high-fat high-sugar (HFHS) diet was used to induce obese models in rats. CB extract was orally gavaged at 0.18, 0.9 and 1.8 g/kg doses for six weeks, and feed intake, body weight, fat pad weight, and blood indexes were measured. Blood serum metabolites were evaluated by gas chromatography/quadrupole time-of-flight tandem mass spectrometry (GC-TOF/MS). Results: The results showed that compared with the obese group, the administration of CB extract caused significant decreases in body weight (P < 0.05), feed intake, Lee’s index, and perirenal, mesenteric, epididymal fat weight. CB extract also reduced blood triglyceride and total cholesterol levels (P < 0.05) of obese rats. Metabolomic findings showed that nine differential metabolites, including pyruvic acid, D-glucuronic acid, malic acid, dimethylglycine, oxoglutaric acid, pantothenic acid, sorbitol acid, fumaric acid and glucose 6-phosphate were identified under CB treatment and altered metabolic pathways such as TCA cycle, pantothenate and CoA biosynthesis, and glycolysis/gluconeogenesis. Conclusion: This study demonstrated weight loss and lipid lowering effects of CB on HFHS diet-induced obese rats and identified nine metabolites as potential biomarkers for evaluating the favorable therapeutic mechanism of CB via regulation of lipid and glucose metabolism.

Metabonomics study of liver and kidney subacute toxicity induced by garidi-5 in rats

2022/9/1 15:43:52

Objective Metabonomics was used to analyze and explore the biomarkers and possible mechanisms of liver and kidney subacute toxicity induced by garidi-5 in rats. Methods Taking garidi-5 as the target drug and SD rats as the research objects, each administration group except the normal group was intragastric administration of the corresponding drug solution for 28 d. The serum, liver and kidney samples of rats were detected by metabolomics and characterized by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) to identify the sensitive markers and metabolic pathways of liver and kidney subacute toxicity. Results Metabolomics analysis showed that compared with the normal group (Z), the 52, 64 and 54 different metabolites were identified in the serum, liver and kidney samples of garidi-5 high dose group (GG), which were mainly enriched in ABC transporters, arginine and proline metabolism, nicotinate and nicotinamide metabolism, central carbon metabolism in cancer pathways. Conclusion The preliminarily suggested that garidi-5 can damage the liver and kidney by affecting the ABC transporters, arginine and proline metabolism, nicotinate and nicotinamide metabolism pathways, etc. Trimethylamine N-oxide, L-pyroglutamic acid, glycine-betaine, xanthine, glutathione, L-leucine, cytidine, L-arginine, spermidine, urea, 5-aminovaleric acid, creatine, L-glutamic acid, 1-methylnicotinamide and S-adenosyl-L-methionine can be used as potential biomarkers of liver and kidney toxicity sensitivity.

Assessment of pulmonary infectious disease treatment with Mongolian medicine formulae based on data mining, network pharmacology and molecular docking

Baochang Zhou a, Zhanhong Qian a, Qinyu Li b, Yuan Gao c,*, Minhui Li a,b,d,e,* — 2022/9/1 15:43:52

Objective: Pulmonary infectious diseases (PID) include viral pneumonia (VP) and pulmonary tuberculosis (PT). Mongolian medicine (MM) is an effective treatment option in China, however, the core group medicines (CGMs) in the treatment of PID and their underlying therapeutic mechanisms remain unclear. In this study, through the method of data mining, the CGMs of MM for the treatment of PID were excavated, and the possible mechanism of action of the CGMs in the treatment of PID was explored by using network pharmacology. Methods: First, 89 MM formulae for the treatment of pulmonary infectious diseases collected from Gan Lu Si Bu, Meng Yi Jin Kui, People's Republic of China Ministry of Health Drug Standards (Mongolian Medicine Volume), Standard of Mongolian Medicine Preparations in Inner Mongolia (2007 Edition), and Standard of Mongolian Medicine Preparations in Inner Mongolia (2014 Edition). The CGMs of MM for PID were excavated through association rule analysis and cluster analysis. Then, the active ingredients and potential targets of the CGMs were obtained from TCMSP, TCMIP, BATMAN-TCM databases. PID targets information was collected from OMIM, GeneCards, and DrugBank databases. The possible targets of CGMs treatment for PID were obtained by intersection. The PPI network was constructed through the STRING database, and the topology analysis of the network was performed. Through the enrichment analysis of the intersection targets by R language, the main action pathways and related target proteins of CGMs in the treatment of PID were screened out. The results were verified by molecular docking. Results: A total of 89 formulae were included, involving 164 MM herbs. The efficacy of the drugs was mainly cough-suppressing and panting-calming herbs, and heat-clearing herbs. The nature and flavor were mainly bitter and cold. The CGMs of MM to treatment of PID was excavated as the classic famous formula Sanzi Decoction (Toosendan Fructus-Chebulae Fructus-Gardeniae Fructus). A total of 28 candidate components and 237 predicted targets of CGMs were collected, and 61 common targets with PID were obtained, including key compounds such as quercetin, kaempferol, β-sitosterol and stigmastero and key targets such as VEGFA, IL6, TP53, AKT1. KEGG enrichment analysis yielded AGE-RAGE signaling pathways, IL-17 signaling pathways, and TNF signaling pathways. Molecular docking results showed that the key targets were well matched with the potential active ingredients of CGMs. Conclusion: This study found that MM commonly used cough-suppressing and panting-calming herbs in combination with heat-clearing herbs to treat PID, and the CGMs for the treatment of PID is “Toosendan Fructus-Chebulae Fructus-Gardeniae Fructus”. CGMs mainly play a role in the treatment of PID by acting on VEGFA, IL6, TP53, AKT1 and other targets, regulating AGE-RAGE signaling pathways, IL-17 signaling pathways, and TNF signaling pathways.

Tea and Citrus maxima complex induces apoptosis of human liver cancer cells via PI3K/AKT/mTOR pathway in vitro

2022/9/1 15:43:52

Objective In this study, black tea and Citrus maxima (BT-CM), yellow tea and C. maxima (YT-CM), green tea and C. maxima (GT-CM) as subjects, the active ingredient content and antioxidant activity of three tea and C. maxima (T-CM) were analyzed. The effects of three T-CMs on apoptosis of liver cells in vitro and its mechanism were further explored. Methods National standard method and HPLC were used for active ingredient analysis. MTT, cell flow cytometry and Western blot were used to analyze the effects of three T-CMs on cell proliferation, apoptosis, and its underlying molecular mechanism. Results The content of tea polyphenols, free amino acids, ratio of polyphenols and amino acids, ester catechins, non-ester catechins and caffeine in YT-CM and GT-CM was significantly higher than that of BT-CM. The in vitro antioxidant capacity of YT-CM and GT-CM was also significantly stronger than that of BT-CM. Three T-CMs had the effects of inhibiting proliferation, arresting cell cycle and inducing apoptosis in HepG2 and Bel7402 cells, especially YT-CM and GT-CM. Western blot analysis showed three T-CMs activated PI3K/AKT/mTOR signaling pathway and regulated the expression levels of apoptosis-related proteins Bax, Bcl-2 and Caspase-3/9. YT-CM and GT-CM had better ability to change the signal pathway than BT-CM. Conclusion In short, T-CMs, which combined different degrees of fermentation tea with C. maxima, were rich in nutrients and biologically active substances. T-CMs, especially YT-CM and GT-CM, are healthy drinks that help to prevent and treat liver cancer.

Chemical constituents of Lomatogonium carinthiacum and Halenia corniculata

2022/9/1 15:43:52

Objective: To study the chemical constituents from traditional characteristic Lomatogonium carinthiacum and Halenia corniculate. Methods: The chemical constituents were isolated and purified by silicagel column, Sephadex LH-20, ODS and high performance liquid chromategramphy. The structures were identified by NMR and MS analysis technics. Results: Twelve compounds were isolated and identified as isovitexin (1), Luteolin-5-O-β-D-glucoside (2), Isosaponarin (3), Luteolin-7-O-β-D-glucoside (4,7), 1,4,8-Trimethoxy-xanthone-6-O-β-D-glucoronyl-(1→6)O-β-Dglucoside (5), friginosideD (6), 1-hydroxy-2,3,5-trimethoxyxanthone (8), 1-hydroxy-2,3,4,5-tetramethoxyxanthone (9), 1-hydroxy-2,3,4,7-tetramethoxyxanthone(10), 1-hydroxy-2,3,4,5,7-pentamethoxyxanthone (11) and usnic acid (12). Conclusion: Compounds 6 and 12 are obtained from this medicine for the first time.

Rapid quantitative analysis of 12 chemical constituents in wild-simulated and cultivated Astragali Radix based on UHPLC-MS

Dan Li a,b, Yuetao Liu a,b,*, Xuemei Qin a,b,* — 2022/9/1 15:43:52

Objective: Astragali Radix (AR) is one of the most widely used traditional Chinese medicines (TCMs) for tonic, which can be divided into wild-simulated and cultivated AR according to its cultivation method. However, whether cultivated AR can replace wild-simulated AR has always been a concern. Methods: In this study, a rapid, highly sensitive and specific analytical method using ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS) was developed to quantitatively measure 12 chemical constituents of AR in the different cultivation methods. Results: AR samples were analyzed with a good linear regression relationship (R2, 0.9983?0.9995), precisions (relative standard deviation (RSD), 1.31%?2.36%), repeatability (RSD, 2.65%?4.92%), stability (RSD, 1.50%?4.05%), and recovery (95.13%?106.52%). Through the determination of AR samples, we found the components of flavonoids in wild-simulated AR were higher than cultivated AR, the saponins in cultivated AR were higher, the ratio of saponins/flavonoids in cultivated AR was higher than wild-simulated AR. Conclusion: Based on this research, it could provide guidance for the quality control of AR.

Asari Radix et Rhizoma consumption lacks relevance for hepatocellular carcinoma in patients: A retrospective cohort study

2022/9/1 15:43:52

Objective: Although some studies have linked Asari Radix et Rhizoma (Asari Radix) administration to hepatocellular carcinoma (HCC), few studies have examined the association between the development of HCC and the use of Asari Radix among patients in mainland China. This study aimed to evaluate the real-world association between Asari Radix and HCC in patients to strengthen the understanding of Asari Radix safety. Methods: A retrospective cohort study among hepatitis B?virus (HBV)-monoinfected patients and non-HBV-monoinfected patients were performed. Patients over 18 years of age were eligible for inclusion. Prescription records of inpatients and outpatients were inquired to distinguish Asari Radix users and nonusers. The risk of developing HCC among Asari Radix users and nonusers in the HBV cohort and the non-HBV cohort was analyzed. Results: There were 49 500 HBV and 133 148 non-HBV patients involved in the two cohorts. Among HBV patients (2 901 users; 46 599 nonusers), the prevalence of HCC in Asari Radix users was lower than that in nonusers (145.70 vs. 265.43 per 105). Among non-HBV patients (5 042 users; 128 106 nonusers), the prevalence of HCC in Asari Radix users was lower than that in nonusers (81.62 vs. 134.11 per 105). None of the hazard ratios (HRs) of Asari Radix exposure ranging from 1 g to 200 g in the two cohorts showed correlation between Asari Radix exposure and hepatocarcinogenesis. Conclusion: An obvious irrelevancy was found between the consumption of Asari Radix and HCC development both in patients with and in those without HBV infection. Use of Asari Radix under 200 g appears safe in terms of HCC risk in the Chinese population; further prospective studies are needed to confirm our results.

6,6′-bieckol induces apoptosis and suppresses TGF-β-induced epithelial-mesenchymal transition in non-small lung cancer cells

Yong-xin Li, Man Liu, Kun-long Yang, Jun Tian — 2022/4/21 13:15:46

Objective: In this study, the aim was to investigate the inhibitory effect of 6,6′-bieckol on the migration and epithelial-mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC) cells, and explore its potential molecular mechanisms. Methods: Cell migration was measured using a CCK8, wound healing, and transwell migration assay. Apoptosis was determined using an Annexin V/propidium iodide staining. Western blotting and immunofluorescence were used to examine the expression level of apoptosis-related proteins and EMT marker proteins. Results: The results showed that 6,6′-bieckol inhibited migration and induced apoptosis of NSCLC cells. Furthermore, 6,6′-bieckol had significantly up-regulated the E-cadherin and down-regulated Snail1 and Twist1 transcriptional levels. 6,6′-Bieckol might inhibit TGF-β-induced EMT by down-regulating Snail1 and Twist1 and up-regulating E-cadherin in lung cancer cells. Conclusion: It was suggested that 6,6′-bieckol has the potential to be developed as a therapeutic candidate for lung cancer.

Extracts of Portulaca oleracea promote wound healing by enhancing angiology regeneration and inhibiting iron accumulation in mice

2022/4/21 13:15:46

Objective: To investigate the role of Portulaca oleracea (POL) in promoting revascularization and re-epithelization as well as inhibiting iron aggregation and inflammation of deep tissue pressure injury (DTPI). Methods: The hydroalcoholic extract of POL (P) and aqueous phase fraction of POL (PD) were prepared based on maceration and liquid-liquid extraction. The number of new blood vessels and VEGF-A expression level were assessed using H&E stain and Western blot on injured muscle to examine the role of POL different extracts in vascularization. The iron distribution and total elemental iron of injured muscle were detected using laser ablation inductively coupled plasma mass spectrometry (ICP-MS) and Perls' stainning to determine whether POL extracts can inhibit the iron accumulation. Besides, the ability of POL extracts to promote wound healing by combining re-epithelization time, inflammation degree and collagen deposition area were comprehensively evaluated. Results: In vitro, we observed a significant increase in HUVEC cell viability, migration rate and the number of the tube after P and PD treatment (P < 0.05). In vivo, administration of P and PD impacted vascularization and iron accumulation on injured tissue, evident from more new blood vessels, higher expression of VEGF-A and decreased muscle iron concentration of treatment groups compared with no-treatment groups (P < 0.05). Besides, shorter re-epithelization time, reduced inflammatory infiltration and distinct collagen deposition were associated with administration of P and PD (P < 0.05). Conclusion: POL extract administration groups have high-quality wound healing, which is associated with increased new blood vessels, collagen deposition and re-epithelization, along with decreased iron accumulation and inflammatory infiltration. Our results suggest that that POL extract is beneficial to repair injured muscle after ischemia-reperfusion, highlighting the potential of POL in the DTPI treatment.

Network pharmacology analysis to explore mechanism of Three Flower Tea against nonalcoholic fatty liver disease with experimental support using high-fat diet-induced rats

2022/4/21 13:15:46

Objective: Nonalcoholic fatty liver disease (NAFLD) has become a common chronic liver disease that is harmful to human health. Moreover, there is currently no FDA-approved first-line drug for the treatment of nonalcoholic steatohepatitis (NASH) or NAFLD. Traditional Chinese medicine (TCM) is widely used to ameliorate liver diseases, such as the traditional ancient recipe called Three Flower Tea (TFT), which consists of double rose (Rosa rugosa), white chrysanthemum (Chrysanthemum morifolium), and Daidaihua (Citrus aurantium). However, the mechanisms of the action of TFT are not clear. Therefore, this study aimed to elucidate the mechanisms of TFT against NAFLD in high-fat diet (HFD)-induced rats. Methods: This study utilized bioinformatics and network pharmacology to establish the active and potential ingredient-target networks of TFT. Furthermore, a protein-protein interaction (PPI) network was constructed, and enrichment analysis was performed to determine the key targets of TFT against NAFLD. Furthermore, an animal experiment was conducted to evaluate the therapeutic effect and confirm the key targets of TFT against NAFLD. Results: A total of 576 NAFLD-related genes were searched in GeneCards, and under the screening criteria of oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18, a total of 19 active ingredients and 210 targets were identified in TFT. Network pharmacology analysis suggested that 55 matching targets in PPIs were closely associated with roles for NAFLD treatment. Through the evaluation of network topology parameters, four key central genes, PPARγ, SREBP, AKT, and RELA, were identified. Furthermore, animal experiments indicated that TFT could reduce plasma lipid profiles, hepatic lipid profiles and hepatic fat accumulation, improve liver function, suppress inflammatory factors, and reduce oxidative stress. Through immunoblotting and immunofluorescence analysis, PPARγ, SREBP, AKT, and RELA were confirmed as targets of TFT in HFD-induced rats. Conclusion: In summary, our results indicated that TFT could prevent and treat NAFLD via multiple targets, including lipid accumulation, antioxidation, insulin sensitivity, and inflammation.

Exploration of Hanshi Zufei prescription for treatment of COVID-19 based on network pharmacology

2022/4/21 13:15:46

Objective: Network pharmacology combines drug and disease targets with biological information networks based on the integrity and systematicness of the interactions between drugs and disease targets. This study aims to explore the molecular basis of Hanshi Zufei formula for treatment of COVID-19 based on network pharmacology and molecular docking techniques. Methods: Using TCMSP, the chemical constituents and molecular targets of Atractylodis Rhizoma, Citri Reticulatae Pericarpium, Magnoliae Officinalis Cortex, Pogostemonis Herba, Tsaoko Fructus, Ephedrae Herba, Notopterygii Rhizoma et Radix, Zingiberis Rhizoma Recens, and Arecae Semen were investigated. The predicted targets of novel coronavirus were screened using the NCBI and GeneCards databases. To further screen the drug-disease core targets network, the corresponding target proteins were queried using multiple databases (Biogrid, DIP, and HPRD), a protein interaction network graph was constructed, and the network topology was analyzed. The molecular docking studies were also performed between the network’s top 15 compounds and the coronavirus (SARS-CoV-2) 3CL hydrolytic enzyme and angiotensin conversion enzyme II (ACE2). Results: The herb-active ingredient-target network contained nine drugs, 86 compounds, and 49 drug-disease targets. Gene ontology (GO) enrichment analysis resulted in 1566 GO items (P < 0.05), among which 1438 were biological process items, 35 were cell composition items, and 93 were molecular function items. Fourteen signal pathways were obtained by enrichment screening of the KEGG pathway database (P < 0.05). The molecular docking results showed that the affinity of the core active compounds with the SARS-CoV-2 3CL hydrolase was better than for the other compounds. Conclusion: Several core compounds can regulate multiple signaling pathways by binding with 3CL hydrolase and ACE2, which might contribute to the treatment of COVID-19.

Functional and binding studies of gallic acid showing platelet aggregation inhibitory effect as a thrombin inhibitor

2022/4/21 13:15:46

Objective: This study was devoted to identifying natural thrombin inhibitors from traditional Chinese medicine (TCM) and evaluating its biological activity in vitro and binding characteristics. Methods: A combination strategy containing molecular docking, thrombin inhibition assay, surface plasmon resonance (SPR) and molecular dynamics simulation were applied to verify the study result. Results: Gallic acid was confirmed as a direct thrombin inhibitor with IC50 of 9.07 μmol/L and showed a significant inhibitory effect on thrombin induced platelet aggregation. SPR-based binding studies demonstrated that gallic acid interacted with thrombin with a KD value of 8.29 μmol/L. Molecular dynamics and binding free energy analysis revealed that thrombin-gallic acid system attained equilibrium rapidly with very low fluctuations, the calculated binding free energies was ?14.61 kcal/mol. Ala230, Glu232, Ser235, Gly258 and Gly260 were the main amino acid residues responsible for thrombin inhibition by gallic acid, providing a mechanistic basis for further optimization. Conclusion: This study proved that gallic acid is a direct thrombin inhibitor with platelet aggregation inhibitory effect, which could provide a basis for the follow-up research and development for novel thrombin inhibitors.

Preparation and pharmacokinetics in vivo of linarin solid dispersion and liposome

2022/4/21 13:15:47

Objective: The current investigation aimed to determine the appropriate dosage form by comparing solid dispersion and liposome to achieve the purpose of improving the solubility and bioavailability of linarin. Methods: Linarin solid dispersion (LSD) and linarin liposome (LL) were developed via the solvent method and the thin film hydration method respectively. The Transwell chamber model of Caco-2 cells was established to evaluate the absorption of drug. The pharmacokinetics of linarin, LSD and LL in rats after ig administration were carried out by high performance liquid chromatography (HPLC) method. Results: The solubility of LSD and LL was severally 3.29 times and 3.09 times than that of linarin. The permeation coefficients of LSD and LL were greater than 10-6, indicating that the absorption of LSD and LL were both better than linarin. The bioavailability of the LSD was 3.363 times higher than that of linarin, and the bioavailability of LL was 0.9886 times higher than that of linarin. Conclusion: The linarin was more suitable for making solid dispersion to enhance its solubility and bioavailability.

Difference analysis of different parts of chicory based on HPLC fingerprint and multi-component content determination

Mengzhen Yana,b, Zhenling Zhanga,b,c,*, Yanze Liud,* — 2022/4/21 13:15:47

Objective: To establish HPLC fingerprints of different parts of chicory stems, leaves, roots, flowers and seeds, and compare the similarities and differences of chemical components in different parts, so as to provide a scientific basis for the comprehensive utilization of chicory. Methods: To establish the HPLC fingerprint of chicory, the chromatographic column was chosen with Agilent ZORBAX Eclipse XDB-C18, the mobile phase was methanol (A) - 0.2% formic acid (B), the flow rate was 1 mL/min, the column temperature was 30 °C, and the detection wavelength was 254 nm. The Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine (2012 Edition) was used to evaluate the similarity of different parts of decoction pieces, and the determination method of multi-component content was established based on fingerprint identification chromatographic peaks, and the determination results were analyzed. Results: The HPLC fingerprinting method of chicory was established. Sixteen chromatographic peaks were identified and 10 of them were identified as: caftaric acid (1), esculin (2), chlorogenic acid (3), esculetin (4), caffeic acid (5), cichoric acid (8), hyperoside (11), rutin (12), isochlorogenic acid C (14) and luteolin (16). The similarity of different parts was 0.084?0.701. At the same time, the total content of detected chemical components was ranked as flower > leaf > stem > root > seed. Roots did not contain caftaric acid, rutin, and luteolin, flowers did not contain luteolin, and seeds did not contain caftaric acid, cichoric acid, and luteolin. The content of cichoric acid in leaves was the most, and esculin in flowers was the most. Conclusion: The results of HPLC fingerprint and multi-component content determination revealed the similarity and difference of different parts of chicory from chemical composition, indicating that there were certain differences in different parts of chicory. The established HPLC fingerprinting method can provide a reference for quality control and evaluation of different parts of the chicory.

Quantitative determination of multi-class bioactive constituents for quality control of Yiqi Jiangzhi Granules

2022/4/21 13:15:47

Objective: To establish a reliable and sensitive method for evaluating quality of Yiqi Jiangzhi Granules (YQJZG). Methods: Ultra performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) was employed for simultaneous determination of eight marker components. Separation was performed on an AQUITY UPLC? HSS T3 column, the mobile phase consisted of acetonitrile as the organic phase and 0.1% (volume percentage) formic acid as the aqueous. Eight marker components, ginsenoside Rg1 (GRg1), ginsenoside Re (GRe), ginsenoside Rb1 (Gb1), typhaneoside (TEO), isorhamnetin-3-O-neohespeidoside (IN), hesperidin (HPD), aurantio-obtusin-6-O-β-D-glucoside (AG) and curcumin (CCM), were detected by multiple reaction monitoring (MRM) mode. The Chinese Pharmacopoeia (2020 edition) was regarded as the guidance document for this method validation. Results: The method showed good linearity (R2 ≥ 0.9990). The relative standard deviation (RSD) values for the instrument precision, intermediate precision and repeatability were less than 2.91%, 2.88%, and 3.54%, respectively. The average recovery varied from 91.08% to 103.89%, with RSD below 3.81%. Sample solutions were found to be stable within 24 h at 4 °C (RSD < 2.85%). Eight marker components were successfully determined from three batches of YQJZG. Conclusion: The proposed UPLC-ESI-MS/MS method was found to be simple, fast and sensitive, and can be used for the routine quality assessment of YQJZG. Simultaneously, this method may provide a new and powerful tool of quality control for other traditional Chinese medicine analogous formulae.

Cathayanalactone G and other constituents from leaves and twigs of Callicarpa cathayana

2022/4/21 13:15:47

Objective: To study the chemical constituents from the leaves and twigs of Callicarpa cathayana. Methods: The chemical constituents were isolated and purified by column chromatography on silica gel, MCI gel CHP 20P/P120, Sephadex LH-20, and HPLC. The structures of the compounds were determined by HR-ESI-MS, 1D and 2D NMR data. Results: A total of 24 compounds were isolated from the 85% methanol extract of leaves and twigs of C. cathayana. They were identified as cathayanalactone G (1), a new diterpene, and 23 known compounds as patagonic acid (2), (-)-16-hydroxycledroda-3,13-dien-16,15-olide-18-oic acid (3), 15-methoxypatagonic acid (4), oleanolic acid (5), ursolic acid (6), siaresinolic acid (7), pomolic acid (8), α-amyrin (9), tormentic acid (10), lupeol (11), 5,7-dihydroxy-3,4'-dimethoxyflavone (12), 5,4'-dihydroxy-3,7,3'-dimethoxyfla-vone (13), 5-hydroxy-3,6,7,4'- tetramethoxyflavone (14), salvigenin (15), kaemferol (16), astragalin (17), pinoresinol 4-O-β-D-glucopyranoside (18), paulownin (19), β-sitosterol (20), β-sitosterol β-D-glucopyranoside (21), 5-hydroxy-coumarin (22), isocopoletin (23), and 4-hydroxycinnamic acid (24). Conclusion: Compound 1 is a new labdane diterpene. Compounds 10, 13, 16 and 17 are isolated from the genus Callicarpa for the first time. Compounds 7, 8, 9, 12, 14, 23 and 24 are reported from C. cathayana for the first time.

Three new triterpenoid saponins from Aralia echinocaulis

Mengyun Yan, Yajia Sun, Ling Ding, Jiuxu Sun, Jingzhu Song, Wenbin Zhou — 2022/4/21 13:15:47

Objective: To study the active ingredients in the root bark of Aralia echinocaulis. Methods: Three triterpenoid saponins were separated from the 70% ethanol extracts and purified by column chromatography and their structures were determined by spectroscopic analysis. Compound 1 and 3 were evaluated for antioxidant activity by the in vitro DPPH free radical scavenging ability and the protective effect of OH- induced DNA oxidative damage. Results: Compound 1 was a new type of triterpenoid saponin, named as echinocaulisaglycone 3-O-β-D-glucopyranoside (echinocaulisaponin A), and it had good antioxidant activity. Compound 2 was similar to compound 1, named as 1-hydroxyl-echinocaulisaglycone 3-O-β-D-glucopyranoside (echinocaulisaponin B). Compound 3 was also a new type of triterpenoid saponin, named as echinocaulisaglycone II 3-O-α-L-arabinopyranosyl-(1”→4’)-β-D-glucopyranosiduronic acid (echinocaulisaponin C), and its antioxidant activity was weaker than compound 1. Conclusion: In this study, three new compounds were discovered and two of them were carried out in vitro anti-oxidation studies, laying the foundation for further research on the treatment of related diseases (cardiovascular disease, arthritis, age-related macular degeneration, etc.) through anti-oxidation or quenching free radical function.

Genome-wide identification and transcriptional profiling analysis of PIN/PILS auxin transporter gene families in Panax ginseng

2022/2/21 14:11:41

Objective: Plant hormones act as chemical messengers in the regulation of plant development and metabolism. The production of ginsenosides in Panax hybrid is promoted by auxins that are transported and accumulated by PIN-FORMED (PIN) and PIN-LIKES (PILS) auxin transporters. However, genome-wide studies of PIN/PILS of ginseng are still scarce. In current study, identification and transcriptional profiling of PIN/PILS gene families, as well as their potential relationship with ginsenoside biosynthesis in Panax ginseng were investigated. Methods: PIN/PILS genes in P. ginseng was identified via in silico genome-wide analysis, followed by phylogenetic relationships, gene structure, and protein profiles investigation. Moreover, previously reported RNA-sequence data from various tissues and roots after infection were utilized for PIN/PILS genes expression pattern analysis. The Pearson’s correlation analysis of specific PIN/PILS genes expression level and main ginsenoside contents were taken to reveal the potential relationship between auxin transports and ginsenoside biosynthesis in P. ginseng. Results: A genome-wide search of P. ginseng genome for homologous auxin transporter genes identified a total of 17 PIN and 11 PILS genes. Sequence alignment, putative motif organization, and sub-cellular localization indicated redundant and complementary biological functions of these PIN/PILS genes. Most PIN/PILS genes were differentially expressed in a tissue-specific manner, and showed significant correlations with ginsenoside content correspondingly. Eight auxin transporter genes, including both PIN and PILS subfamily members, were positively correlated with ginsenoside content (cor > 0.60; P-value < 0.05). The expression levels of eleven auxin transporter genes were increased dramatically in the early stage (0?0.5 DPI) after Cylindrocarpon destructans infection, accompanied with various overall expression patterns, implying the dynamic auxin transport in response to biotic stress. Conclusions: Based on the results, we speculate that the accumulation or depletion in temporal or spatial manner of auxin by PIN/PILS transporters involved in the regulation of HMGR activity and subsequent ginsenoside biosynthesis.

Microbial inoculants and garbage fermentation liquid reduced root-knot nematode disease and As uptake in Panax quinquefolium cultivation by modulating rhizosphere microbiota community

Pei Caoa, Xue-min Wei a, Gang Wanga, Xiao-chen Chen, Jian-ping Hana, Yuan Lib — 2022/2/21 14:11:41

Objective: To find a suitable ecological cultivation measure to solve the problem of root-knot nematode disease of Panax quinquefolium (Panacis Quinquefolii Radix) and the heavy metals accumulating in its roots. Methods: Three-year-old P. quinquefolium was treated with four different combinations of microbial inoculant (MI) and garbage fermentation liquid (GFL) [the joint application of ‘TuXiu’ MI and Fifty potassium MI (TF), the combination use of ‘No. 1’ MI and Fifty potassium MI (NF), ‘Gulefeng’ poly-γ-glutamic acid MI (PGA), GFL], and the untreated control (CK). Here, high-throughput sequencing, ICP-MS and UPLC were employed to systematically characterize changes of microbial diversity and structure composition, heavy metals (As, Cd and Pb) content and ginsenoside content among different treatments. Results: The results revealed that different MIs and GFL could increase the root dry weight of P. quinquefolium, PGA enhanced it by 83.24%, followed by GFL (49.93%), meanwhile, PGA and GFL were able to lessen root-knot nematode disease incidence by 57.25% and 64.35%. The treatment of PGA and GFL can also effectively reduce heavy metals in roots. The As content in GFL and PGA was decreased by 52.17% and 43.48% respectively, while the Cd and Pb contents of GFL and PGA was decreased somewhat. Additionally, the content of total ginsenosides was increased by 42.14% and 42.07%, in response to TF and NF, respectively. Our metagenomic analysis showed that the relative abundance of particular soil microbial community members related to the biocontrol of root-knot nematode disease and plant pathogen (i.e., Chaetomium in NF, Xylari in GFL, and Microascus in PGA), heavy metal bioremediation (Hyphomacrobium in PGA and Xylaria in GFL), and nitrogen fixation (Nordella and Nitrospira in TF) was significantly increased; notably, potential harmful microflora, such as Plectosaphaerella and Rhizobacter, were more abundant in the control group. Conclusion: MI and GFL could improve the quality of P. quinquefolium by modifying its rhizosphere microbial community structure and composition, both of them are beneficial to the development of ecological cultivation of P. quinquefolium.

Didymin attenuates doxorubicin-induced cardiotoxicity by inhibiting oxidative stress

2022/2/21 14:11:41

Objective: This study was designed to investigate the protective effects of didymin (Did) on doxorubicin (DOX)-induced cardiotoxicity. Methods: After pretreatment with Did (2, 4, 8 mg/kg intraperitoneal i.p.) for 7 d, the male C57 mice were injected with single dose of DOX (20 mg/kg i.p.). The cardioprotective effect of Did was observed on the 7th day after DOX treatment. Results: DOX delayed body growth and caused cardiac tissue injury, oxidative stress, and mitochondrial dysfunction. Similar experiments in H9C2 cardiomyocytes showed that DOX reduced cell viability, increased generation of reactive oxygen species (ROS) and fragmentation of DNA, decreased mitochondrial membrane potential, and induced cardiomyocyte apoptosis. However, all of these adverse effects were suppressed by Did pretreatment. Did increased protein expression of glutamate-L-cysteine ligase catalytic subunit (GCL), heme oxygenase 1 (HO-1), and nuclear factor erythroid 2-related factor 2 (Nrf2). Besides, Did also induced activation of PI3K/AKT. Conclusion: These findings indicated Did prevented DOX-induced cardiac injury and apoptosis via activating PI3K/AKT/Nrf2 signaling pathway.

He-Wei Granule enhances anti-tumor activity of cyclophosphamide by changing tumor microenvironment

2022/2/21 14:11:41

Objective: He-Wei Granule (HWKL) is a modern product derived from the modified formulation of traditional Chinese medicine Banxia Xiexin Decoction (BXD), which remarkedly enhanced the anti-proliferation activity of cyclophosphamide (CTX) on HepG2 and SGC-7901 cell lines in vitro in our previous research. The aim of the study was to investigate the synergistic effects of HWKL and CTX using a transplanted H22 hepatocellular carcinoma mouse model. Methods: The CTX-toxic-reducing efficacy of HWKL was evaluated by hematology indexes, organ indexes and marrow DNA detection. To investigate the underlying mechanisms, histopathology test, immunohistochemistry test and TUNEL staining were conducted. The efficacy of HWKL on the micro-vessel density (MVD) in tumor tissue was also evaluated by measuring CD34 level. Results: High dose HWKL (6.75 g/kg) markedly attenuated CTX-induced hepatotoxicity and myelosuppression while significantly enhanced CTX anticancer efficacy in vivo. Further mechanism investigation suggested that high dose HWKL significantly increased cleaved Caspase 3 level and promoted apoptosis in tumor tissue by up-regulating Bax expression and down-regulating Bcl-2 and FasL expressions. Compared with CTX alone group, the decrease in LC-3B and Beclin 1 levels suggested that the autophagy in H22 carcinoma was significantly inhibited with addition of high dose HWKL. ELISA assay results indicated that the autophagy inhibition was achieved by decreasing p53 expression, blocking PI3K/AKT/mTOR pathway and recovering Th1/Th2 cytokine balance. In addition, CD34 and EGFR immunohistochemistry assay suggest that high dose HWKL could significantly decrease micro-vessel density (MVD) and inhibit angiogenesis in H22 carcinoma. Conclusion: It could be concluded that high-dose HWKL enhanced CTX efficacy by promoting apoptosis, inhibiting autophagy and angiogenesis in tumor tissue while significantly alleviated CTX-induced toxicity, and could be applied along with CTX in clinical treatment as a supplement agent.

Immunomodulatory activity of polysaccharides from Brassica rapa by activating Akt/NF-ÎºB signaling

2022/2/21 14:11:41

Objective: To investigate the immunomodulatory activity of polysaccharides from the roots of Brassica rapa. Methods: The crude polysaccharide from roots of B. rapa (BRP) was extracted and purified to further investigate the active fraction of BRT for inducing macrophage phagocytosis. Results: Effects on RAW264.7 cells demonstrated that BRP behaved better phagocytic capacity and had potent immunomodulatory activity, including increasing production of nitric oxide (NO), tumor necrosis factor α (TNFα) and upregulating mRNA levels of inducible NO synthase (iNOS) and TNFα. Furthermore, modulation of macrophage by BRP was indicated to be mediated via the activation of Akt and nuclear factor-kappa B (NF-κB). Conclusion: The beneficial effects of BRP could be used as an immunotherapeutic adjuvant in treatment of inflammatory diseases.

Anti-diabetic effects of linarin from Chrysanthemi Indici Flos via AMPK activation

2022/2/21 14:11:41

Objective: The purpose of this study is to investigate the anti-diabetic effects of linarin, a flavonoid extracted from Chrysanthemi Indici Flos (CIF), and its potential mechanisms. Methods: The effects of linarin on cell viability and glucose consumption in HepG2 cells were measured. Meanwhile, monosodium glutamate (MSG) mouse model was constructed to monitor the changes of insulin tolerance, glucose tolerance, triglyceride and cholesterol. The protein expression levels of p-AMPK, p-ACC, PEPCK and p-GS were detected by Western blot. Results: Linarin could increase the relative glucose consumption of HepG2 cells, improve insulin tolerance and glucose tolerance, and decrease the levels of triglyceride and cholesterol of MSG mice. Simultaneously, the expression levels of p-AMPK and p-ACC in HepG2 cells and the liver tissue of MSG mice were increased, while the expression levels of PEPCK and p-GS were decreased after treatment with linarin. Conclusion: Insulin resistance could be ameliorated by linarin in type 2 diabetes, and its mechanism may be related to AMPK signaling pathway.

Effects of novel Fufang Biejia Ruangan Tablets with sheep placenta as substitute for hominis placenta on CCl4-induced liver fibrosis

2022/2/21 14:11:41

Objective Fufang Biejia Ruangan Tablet (FBRT) is widely used for the treatment of liver fibrosis. However, Hominis Placenta (HP), as an important adjuvant of FBRT, has been restricted for medicinal using due to the limited availability, ethical controversy and safety issues. The present study aimed to investigate the therapeutic effects of novel FBRT (N-FBRT) with sheep placenta (SP) as substitute for HP on liver fibrosis and explore its possible mechanisms. Different dosages of SP in N-FBRT were also evaluated. Methods Rats were subcutaneously injected with CCl4 to induce liver fibrosis and then treated with N-FBRT and FBRT. The anti-hepatic fibrosis effect was determined based on biomarkers analysis of liver function and hepatic fibrosis, and the liver pathology was visualized by H&E staining and Masson staining. The oxidative stress and inflammatory cytokines were also detected. Immunohistochemical staining of α-SMA, real time PCR and Western blotting were performed to evaluate hepatic stellate cells (HSCs) activation and TGF-β1/Smad signaling pathway. Results N-FBRT and FBRT could ameliorate CCl4-induced liver fibrosis and improve liver function, as evidenced by lowering serum biomarkers levels of liver function and hepatic fibrosis, and decreasing hepatic Hyp content and collagen deposition, and improving the hepatic morphology and architecture changes. Moreover, the anti-liver fibrosis effect was better when the dosage of SP used in N-FBRT was 1/2 of HP in FBRT. Administration of N-FBRT markedly alleviated oxidative stress and inflammatory cytokines, and inhibited α-SMA expression. Furthermore, the mRNA expression of Col I, Col III, α-SMA and TGF-β1, and proteins expression of α-SMA, TGF-β1, Smad2/3 and p-Smad2/3 were significantly down-regulated by N-FBRT treatment. Conclusion SP can be used as substitute for HP to prepare N-FBRT for the treatment of liver fibrosis and the anti-liver fibrosis effect of N-FBRT is achieved by eliminating oxidative stress and inflammation, and inhibiting HSCs activation and ECM production by blocking TGF-β1/Smad signaling pathway.

Capilliposide from Lysimachia capillipes promotes terminal differentiations and reverses paclitaxel resistance in A2780T cells of human ovarian cancer by regulating Fos/Jun pathway

Ke Zhanga, Han-yue Yingb, Ru-ping Zhaoa, Yuan-yuan Chena, Qing-hua Denga — 2022/2/21 14:11:41

Objective: To investigate the potential effect of Lysimachia capillipes capilliposide (LCC) on the chemo sensitivity and the stemness of human ovarian cancer cells. Methods: Cell Counting Kit-8 (CCK8) was used to measure the IC50 values. The apoptosis of cells was measured through flow cytometry. Evaluation of the stemness and differentiation markers was performed by the immunoblotting and the immunostaining assays. RNA-seq was performed through the Illumina HiSeq PE150 platform and differentially expressed genes (DEGs) were screened out through the bioinformation analysis. Overexpression or knockdown of Fos gene was achieved by shRNA transfection. Results: Pre-exposure of A2780T cells with 10 μg/mL LCC sensitized them to paclitaxel, of which the IC50 value reduced from 8.644 μmol/L (95%CI: 7.315?10.082 μmol/L) to 2.5 μmol/L (95%CI: 2.233?2.7882 μmol/L). Exposure with LCC enhanced the paclitaxel-induced apoptosis and inhibited the colony formation of A2780T cells. LCC exposure reduced the expression of cancer stemness markers, ALDH1, Myd88 and CD44, while promoting that of terminal differentiation markers, NFATc1, Cathepsin K and MMP9. RNA-seq analysis revealed that the expressions of FOS and JUN were upregulated in LCC-treated A2780T cells. A2780T cells overexpressing Fos gene displayed increased paclitaxel-sensitivity and reduced cell stemness, and shared common phenotypes with LCC-treated A2780T cells. Conclusion: These findings suggested that LCC promoted terminal differentiations of ovarian cancer cells and sensitized them to paclitaxel through activating the Fos/Jun pathway. LCC might become a novel therapy that targets at cancer stem cells and enhances the chemotherapeutic effect of ovarian cancer treatments.

Effect and molecular mechanism research of Astragalus membranaceus on inhibiting intestinal absorption of six alkaloids of Aconitum carmichaelii in spleen deficiency rats

2022/2/21 14:11:41

Objective To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods Four-week-old Male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.

Analgesic alkaloids from Urticae Fissae Herba

Xiao-ru Fenga, Pei-jun Jub, Yan Chena, Xiao-bo Lia, Meng-yue Wanga — 2022/2/21 14:11:41

Objective: To investigate the analgesic substances in the aerial part of Urtica fissa (Urticae Fissae Herba), commonly used for rheumatoid and rheumatism arthritis. Methods: The analgesic constituents were isolated with the active guidance of hot plate and acetic acid writhing models, and identified by comprehensive spectroscopic analysis. Results: Thirteen alkaloids (1?13), two lignans (14, 15), and three amides (16?18) were isolated from the active fractions. Among them, compound 1 was a new alkaloid, and compound 6 was a new natural product. The activity evaluation in vivo indicated that various pyrrole alkaloids (1, 3, 6, and 12) possessed significant analgesic activities, they could significantly inhibit the mice pain response induced by acetic acid and hot plate at the dosage of 2 mg/kg BW. Conclusion: The study revealed that the pyrrole alkaloids played important roles in the analgesic activities of Urticae Fissae Herba.

Xiaoke Decoction in treatment of type II diabetes: A Meta-analysis

2022/2/21 14:11:41

Objective: To systematically evaluate the clinical effect of Xiaoke Decoction in the treatment of type 2 diabetes. Methods: Chinese databases such as CNKI, Wanfang, Weipu Chinese Biomedical Journal Database, and Chinese Medical Biological Literature Database, PubMed, Cochrane Library, Embase, and Web of Science were searched for english language literature from their inception until November 2019. A Meta-analysis was performed using RevMan 5.3 and Stata 12.0. Results: Thirty-eight studies were included in this study, with a total of 3757 patients. It was found that adding Xiaoke Decoction could improve total efficiency. The Xiaoke Decoction groups surpassed the western medicine groups regarding improvement in total efficiency (OR=3.49; 95% CI: 2.78–4.39, P < 0.00001). Adding Xiaoke Decoction could lower the fasting plasma glucose (FPG) level. The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in FPG levels (MD=-1.14; 95% CI: -1.36－-0.92, P < 0.00001). Adding Xiaoke Decoction could lower the 2h postprandial blood glucose (2hPG) level. The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in 2hPG (MD=-1.40; 95% CI: -1.61－-1.19, P < 0.00001). Adding Xiaoke Decoction could lower glycated hemoglobin (HbA1c). The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in HbA1c (MD=-0.77; 95% CI: -0.95－-0.58, P < 0.00001). It was found that adding Xiaoke Decoction could lower the traditional Chinese medicine (TCM) syndrome score. The TCM syndrome scores among patients in the Xiaoke Decoction group were lower than those among patients in the control group after treatment (MD=-4.90; 95% CI: -7.22－-2.57, P < 0.0001). At the same time, we conducted a subgroup and sensitivity analysis of age and intervention duration on the heterogeneity of total efficiency, FPG, 2hPG, HbAlc, and TCM syndrome score outcome indicators. For detecting publication bias, an egger test was conducted. Conclusion: Compared with western medicine alone, Xiaoke Decoction has more advantages for the treatment of type 2 diabetes with respect to total efficiency, FPG, 2hPG, HbAlc, and TCM syndrome score.

Systematic review and Meta-analysis of 26 randomized controlled clinical trials of Compound Danshen Dripping Pill for non-proliferating diabetic retinopathy

Yu Wang, Lei Hao, Zhi-peng Huo, Yuan-xue Liu, Yu-jing Sun, Zhao-hui Song — 2022/2/21 14:11:41

Objective: Diabetic retinopathy (DR) is the retinal consequence of chronic progressive diabetic microvascular leakage and occlusion. Non-proliferating diabetic retinopathy (NPDR) is the early stage of DR. It eventually occurs to some degree in all patients with diabetes mellitus. In recent years, many clinical trials have shown that Compound Danshen Dripping Pill (CDDP) may be associated with the improvement of NPDR symptoms. The aim of this study was to quantitatively summarize the association between CDDP and the therapeutic effects of NPDR. Methods: It was conducted that a systematic literature search of PubMed, Web of Science, CNKI, VIP and Wanfang Data updated in June 2020 with the following search terms: “diabetic retinopathy” or “retinopathy” or “DR” or “NPDR”, in combination with “Compound Danshen Dripping Pill” or “Salvia miltiorrhiza” or “Danshen”. Risk ratio (RR) and weighted mean difference (WMD) with their 95% confidence interval (CI) was calculated between treatment and control groups. The sensitivity analyses were undertaken by removing each individual study when high heterogeneity appeared. Subgroup analysis, Meta-regression, and publication bias analysis were also conducted. The strength of evidence was evaluated with the Grading of Recommendation, Assessment, Development, and Evaluation (GRADE) method. Results: Twenty-six RCTs involving 2047 subjects were included to conduct a Meta-analysis after screening the studies, extracting the data, and assessing the study quality. The Stata15.0 software was utilized for processing. Meta-analysis indicated that curative effects of treatment group with CDDP was significantly better than control (RR=0.54, 95% CI (0.40, 0.73); moderate-quality evidence). In addition, the results showed that CDDP was significantly associated with improving retinal hemorrhages (WMD=?0.62, 95% CI (?0.78, ?0.46); low-quality evidence), the vision (WMD=0.14, and 95% CI (0.09, 0.19), low-quality evidence), fundus fluorescence angiography (RR=0.37 and 95% CI (0.23, 0.60); low-quality evidence), reduction of retinal microaneurysm (WMD=-3.74 and 95% CI (?4.38, ?3.11); moderate-quality evidence), hemangioma volume (WMD=?3.15, 95%CI (?3.45, ?2.85); moderate-quality evidence), macular thickness (WMD=?5.52, 95%CI=(?64.27, ?48.78); low-quality evidence), mean defect (WMD=?1.65 and 95% CI (?1.95, ?1.34); very low-quality evidence), fasting blooding glucose (WMD=?0.95, 95% CI (?1.19, ?0.70); low-quality evidence), hemoglobin A1c (WMD=?0.62, 95% CI (?0.93, ?0.30); low-quality evidence), high sensitive C reaction protein (WMD=?5.66, 95% CI (?8.01, ?3.31); low-quality evidence). Sensitivity, subgroup, and meta-regression analyses were also assessed. Conclusion: The study demonstrated that CDDP has beneficial clinical effects for treating NPDR and improve the vision. Moreover, it indicated that oral CDDP in NPDR patients led to significant regulation of serum level of fasting blooding glucose, hemoglobin A1c and high sensitive C reaction protein, which was associated with the pathogenesis of NPDR. However, high-quality and large randomized clinical trials will be needed to prove the consequence in future.

Safety and efficacy of Compound Huangdai Tablets combined with all-trans retinoic acid for treatment of acute promyelocytic leukemia: Clinical evidence and potential mechanisms

2022/2/21 14:11:42

Objective: To evaluate the safety and efficacy of Compound Huangdai Tablets (Realgar-Indigo Naturalis formula, RIF) combined with all-trans retinoic acid (ATRA) to treat acute promyelocytic leukemia (APL). Methods: This study was registered in PROSPERO (CRD42018108118). The relevant literatures on RIF treatment of APL were systematically searched in the following databases: China National Knowledge Infrastructure, Wanfang, VIP Medical Information System, Chinese Biomedical Database, EMBASE, Cochrane Library, and PubMed. The quality of the included studies was evaluated and Review Manager 5.3 software and Stata 13.0 software were used to perform the Meta-analysis. In addition, this study will use the method of network pharmacology to conduct a preliminary exploration of the mechanism of RIF on APL. Results: The study included 12 studies involving totally of 775 APL patients. The Meta-analysis showed that there was no significant difference (P ＞ 0.05) between the RIF group and the arsenic trioxide (ATO) group for primary outcomes, secondary outcomes apart from liver dysfunction. The incidence of liver dysfunction (P = 0.006) in the RIF group were significantly lower than those in the ATO group. In addition, the cost of maintenance therapy in the RIF group was significantly lower (P ＜ 0.05) than the ATO group. Besides, the active ingredients in RIF mainly act on targets proteins such as ACHE, NCOA2, RXRA, then regulate TP53 Regulates Transcription of Cell Cycle Genes, nuclear receptor transcription pathway and other ways to play a role in treating APL. Conclusion: There was no significant difference in efficacy of oral RIF combined with ATRA compared with intravenous ATO combined with ATRA for the treatment of APL. The oral RIF exposed patients to less risk, offered more convenience and had lower prices. RIF can treat APL by multi-target and multi-pathway interventions that inducing apoptosis of APL cells and inhibiting the proliferation of APL cells, and so on. Therefore, oral RIF in the treatment of APL is worthy of further research and development.

Isolation, characterization and modulatory potentials of β-stigmasterol, ergosterol and xylopic acid from Anchomanes difformis on mitochondrial permeability transition pore in vitro

2023/11/9 13:22:26

Objective: Secondary metabolites and polyphenolic compounds from medicinal plants have been demonstrated to have multiple biological functions with promising research and development prospects. This study examined the effect of β-stigmasterol (with ergosterol) and xylopic acid isolated from Anchomanes difformis on liver mitochondrial permeability transition pore (mPTP). Methods: The compounds were isolated by vacuum liquid chromatography. Mitochondrial swelling was assessed as changes in absorbance under succinate-energized conditions. Results: 1H and 13C NMR spectroscopic elucidation of the isolates affirmed the presence of β-stigmasterol with ergosterol (1:0.3) and xylopic acid. The isolates reversed the increase in lipid peroxidation and inhibited the opening of mitochondrial permeability transition pores caused by calcium and glucose. Pharmacological inhibition of mPTP offers a promising therapeutic target for the treatment of mitochondrial-associated disorders. Conclusion: Reduction in the activity of calcium ATPase and the expression of Caspase-3 and-9 were observed, suggesting that they could play a role in protecting physicochemical properties of membrane bilayers from free radical-induced severe cellular damage and be useful in the management of diseases where much apoptosis occurs.

Scutellarin alleviates liver injury in type 2 diabetic mellitus by suppressing hepatocyte apoptosis in vitro and in vivo

2023/11/9 13:22:26

Objective: Scutellarin is a primary active composition come from Erigeron breviscapus. It is well known that scutellarin has anti-inflammatory and antioxidant physiological functions. In this study, we detect the effects of scutellarin on hepatocyte cell apoptosis in type 2 diabetes mellitus (T2DM) rats. Methods: Sprague Dawley (SD) (6–8 weeks, 160–180 g) rats were randomly divided into six groups: control, model, scutellarin low-dose, medium-dose, high-dose treatment, and rosiglitazone positive groups; with 10 SD rats in each group (n = 10). The changes of biochemical factors in serum were detected by automatic biochemical instrument, the pathological changes of liver tissue were detected by hematoxylin and eosin (HE) staining, the apoptosis of liver tissue and cells was detected by tissue staining and flow analyzer, and the expression of apoptosis-related factors were determined by qPCR, Western blot and immunohistochemistry in liver tissues or cells. Results: The results showed that scutellarin decreased the levels of fasting blood glucose, total cholesterol, triglyceride, and low-density lipoprotein and increased the levels of high-density lipoprotein. Meanwhile, scutellarin decreased the levels of alanine transaminase (ALT) and aspartate transaminase (AST) and improved liver function. In addition, scutellarin suppressed the secretion of interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and reduced hepatocyte apoptosis. Furthermore, scutellarin inhibited the expression of cleaved Caspase-3, Bax, and cytochrome C (Cyt-C) and promoted the expression of Bcl-2. Conclusion: Scutellarin can inhibit the apoptotic pathway, thereby relieving T2DM.

Antioxidant and antiarthritic potential of berberine: In vitro and in vivo studies

Shweta Jain a, Shalini Tripathi b, Pushpendra Kumar Tripathi c,* — 2023/11/9 13:22:26

Objective: To extract and isolate berberine from Berberis aristata (Berberidaceae). Isolated berberine was characterised using spectroscopy and its antioxidant and antiarthritic activity was analyzed. Methods: The berberine was isolated from B. aristata using microwave-assisted extraction (MAE) and characterised by a spectroscopic technique. The isolated berberine was evaluated for its antioxidant activity in DPPH, nitric oxide, and superoxide scavenging assays, while antiarthritic activity was evaluated in the complete freund’s adjuvant (CFA)-induced arthritis rat model. Results: The antioxidant activity of berberine revealed potent antioxidant activity in DPPH, nitric oxide, and superoxide scavenging assays. The in vivo antiarthritic activity of berberine in the CFA-induced arthritis rat model showed a significant reduction in paw diameter, arthritic score, and an increase in body weight. Furthermore, a concentration-dependent ameliorating action of berberine on haematological parameters was noticed. Proinflammatory biomarkers, including IL-6, IL-10, and TGF-b in serum were reported, and histopathology examination revealed that berberine decreased pannus formation, synovial hyperplasia, and bone erosion. Radiographic investigation showed soft tissue inflammation, bone resorption and erosion, joint gap reduction, and substantial connective tissue expansion after treatment with berberine. Conclusion: The ameliorating action on haematological parameters and proinflammatory biomarkers of berberine makes them a suitable remedy for the treatment of arthritis.

Mangiferin alleviates renal inflammatory injury in spontaneously hypertensive rats by inhibiting MCP-1/CCR2 signaling pathway

2023/11/9 13:22:26

Objective: Hypertension is a low-grade inflammation state of the disease and was easily complicated by kidneys’ inflammatory response. Mangiferin (MGF), a pharmacologically active compound in various plants including Mangifera indica, has a strong anti-inflammatory activity. However, the effects of MGF on renal inflammatory injury in spontaneously hypertensive rats (SHRs) remain unclear. The purpose of this study was to investigate the protective effects and mechanisms of MGF on renal inflammatory injury in SHRs. Methods: MGF was used in SHRs at the doses of 10, 20, 40 mg/kg/d for 8 weeks consecutively. The blood and urine were collected for assessment of renal function. Renal tissues were collected for histological, immunohistochemistry, ELISA, Western blot and real time reverse transcription PCR (RT-PCR) analysis. Results: The results showed that the levels of interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and recombinant chemokine C-C-Motif receptor 2 (CCR2) were increased in SHRs, meanwhile, the level of IL-10 was decreased in SHR. Treatment of MGF inhibited the expression of IL-6, TNF-a, MCP-1 and CCR2, and promoted the expression of IL-10. Furthermore, the content of blood urea nitrogen (BUN) and serum uric acid (SUA) was significantly increased in the model group, and treatment of MGF had no obvious effects on these parameters at all dose levels. Conclusion: Our study proved that the kidneys of SHRs had significant inflammatory injury, and MGF had the protective effects on renal inflammatory injury in SHRs; The protective mechanism may be mediated partly by the MCP-1/CCR2 signaling pathway. Thus, it is a potential new drug for the treatment of hypertension.

Modulative effect of Physalis alkekengi on both gut bacterial and fungal micro-ecosystem

Yanan Yang a, Xiaohui Zhao b, Yong Xie a,*, Chongming Wu b,* — 2023/11/9 13:22:26

Objective: Gut microbiome is an intricate micro-ecosystem mediating the human health and drug efficacy. Physalis alkekengi (PAL) is an edible and time-honored traditional Chinese medicine. Several pharmacological effects of PAL have been verified and gut bacteria are implied in its therapeutic actions. However, the detailed modulation of PAL on gut bacterial species and on gut fungi remains largelym unknown. We, therefore, designed a preliminary experiment in normal mice to reveal the modulation effect of PAL on both gut bacteria and fungi, and explore the interaction between them. Methods: Herein, the aqueous extract of PAL was orally administrated to normal C57BL/6 mice for four weeks. The full-length 16S rRNA and ITS1/2 gene sequencing were explored to detect the taxa of gut bacteria and gut fungi after PAL treatment, respectively. Results: Oral administration of PAL notably enriched anti-inflammatory bacterial species such as Duncaniella spp. and Kineothrix alysoides, whereas decreased pro-inflammatory species such as Mucispirillum schaedleri. Simultaneously, PAL increased the abundance of gut fungi Aspergillus ochraceus, Cladosporium sp. and Alternaria sp., and decreased Penicillium janthinellum. Correlation network analysis identified two co-existing microbial groups (groups 1 and 2) that were negatively associated with each other. The group 1 comprised PAL-enriched bacteria and fungi, while group 2 was mainly normal chow-enriched bacteria and fungi. In group 1, Antrodia monomitica, Aspergillus clavatus, Mortierella kuhlmanii and Sarcinomyces sp. MA 4787 were positively correlated with Bifidobacterium globosum, Romboutsia ilealis and so on. In group 2, Chaetomium subspirilliferum, Septoria orchidearum and Cephaliophora tropica were positively related to Lactobacillus spp. Conclusion: Altogether, this preliminary study first demonstrated the modulation effect of PAL on both gut bacteria and gut fungi, which may shed light on the elucidation of PAL’s pharmacological mechanism.

Transcriptome analysis of Saposhnikovia divaricata and mining of bolting and flowering genes

2023/11/9 13:22:26

Objective: Early bolting of Saposhnikovia divaricata has seriously hindered its medicinal value and sustainable development of resources. The molecular mechanism of bolting and flowering of S. divaricata is still unclear and worth of research. In our study, we explored the transcriptome of the genes related to the bolting and flowering of S. divaricata. Methods: The transcriptome library was constructed, sequenced, assembled and annotated from the bolting and unbolting leaves of S. divaricata by high-throughput sequencing at the bud and flowering stage. Focus on the pathways related to bolting and flowering in plants, and explore genes. The expression of seven candidate genes was verified by real-time fluorescence quantitative PCR (qRT-PCR). Results: Transcriptome results showed that 249 889 422 high-quality clean reads were obtained. A total of 67 866 unigenes were assembled with an average length of 948.1 bp. Trinity de Novo assembly produced 67 866 unigenes with an average length of 948.1 bp. Among 993 differentially expressed genes, 484 genes were significantly up-regulated and 509 genes were down-regulated in the SdM group. A total of 79 GO terms were significantly enriched for differentially expressed genes. KEGG results showed that 11 154 unigenes were enriched in 89 pathways. And 21 candidate genes related to bolting and flowering of S. divaricata were excavated. The qRT-PCR results showed that expression trends of HDA9, PHYB, AP2, TIR1, Hsp90, CaM, and IAA7 were consistent with transcriptomic sequencing results. In addition, RNA-seq had identified 10 740 transcription factors and classified them into 58 families by their conserved domains. Further studies showed that the transcription factors regulating the flowering of S. divaricata were mainly distributed in the NAC, MYB_related, HB-other, ARF, and AP2 families. Conclusion: Based on the results of this study, it was found that the plant hormone signal transduction pathway was one of the decisive factors to control bolting and flowering. Among them, auxin related genes IAA and TIR1 are the key genes in the bolting and flowering process of S. divaricata.

Chinese yam yield is affected by soil nutrient levels and interactions among N, P, and K fertilizers

Yang Chen a,1, Xianzhi Zhou a,b,1,*, Lina Ma a, Yongsheng Lin a, Xiangui Huang a — 2023/11/9 13:22:26

Objective: The current fertilization methods for Chinese yam are uneconomic and unfriend to environment. A rational one is very important to achieve desired balance of high yield of Chinese yam, economic and friend to environment. Here, we studied the effects of nitrogen (N), phosphorus (P), and potassium (K) fertilizers on the yield of ‘Qinfeng’ Chinese yam in shallow-groove directional cultivation. Methods: The experiments were conducted in Dehua County, Fujian Province, China using a ‘‘3414” optimal design. Overall, three fertilizer factors (N, P, and K) were evaluated at the following four levels: 0, no fertilizer; 1, 0.5-fold the typical rate; 2, typical fertilization rate; and 3, 1.5-fold the typical rate. There were 14 different fertilization treatments. Results: Treatment 6 (N2P2K2) produced the longest (75.6 cm) and thickest tubers (4.9 cm) with the highest tuber fresh weight (1311.9 g) and yield (41,015.9 kg/hm2), whereas, treatment 1 produced the shortest (65.6 cm) and thinnest tubers (3.9 cm) with the lowest fresh weight (953.4 g) and yield (28 532.8 kg/hm2) among the 14 fertilizer combinations. The experimental data could be fitted to single-variable quadratic and binary quadratic models but not to a ternary quadratic polynomial model. Appropriate N, P, and K fertilizer application rates increased Chinese yam yield. However, excessive fertilization lowered the yield. Chinese yam yield was significantly and strongly correlated with the amounts of N, P, and K fertilizer applied. Conclusion: Based on the single variable quadratic and binary quadratic models, we propose that the quantities of N, P, and K fertilizer used to grow 1 hm2 ‘Qinfeng’ Chinese yam should be 360–388.3, 90–100.95, and 416.3–675 kg, respectively.

Production of hispidin polyphenols from medicinal mushroom Sanghuangporus vaninii in submerged cultures

2023/11/9 13:22:26

Objective: The medicinal mushroom Sanghuangporus vaninii produces pharmaceutically valuable hispidin polyphenols in natural habitats. However, due to the slow growth in nature, S. vaninii grown in the field (sclerotia) is not reliable for pharmaceutical purposes. Although higher biomass of fungal mycelia can be obtained in submerged cultures, the accumulation of hispidin polyphenols is rare. Methods: In this study, the polyunsaturated fatty acids (PUFAs), linoleic acid (LA), linolenic acid (ALA), and methyl jasmonate (MeJa) were employed as the stimulant agents to coordinate the accumulation of biomass and hispidin polyphenols in its submerged cultures. Results: The addition of LA and ALA promoted the mycelial accumulation, while the addition of MeJa inhibited the growth of S. vaninii concomitant with reduced total polyphenols. UPLC-Triple-TOF-MS analysis revealed an increased production of hispidin, phellinstatin, pinnilidine, and its derivatives upon the addition of LA and ALA, and hypholomine B and its isomer, 3,14’-bihispidinyl, and phelligridin E upon the addition of MeJa on day 13. Intriguingly, total polyphenols from the MeJa-supplementing cultures harbored a high capacity in scavenging free radicals. Chemical structural analysis showed that hispidin polyphenols had higher antioxidant activity due to more hispidin moieties induced by MeJa. Conclusion: The supplement of PUFAs affects the synthesis and composition of hispidin polyphenols in S. vaninii. Our results provide a possibility to coordinate the production of hispidin polyphenols via submerged cultures of S. vaninii.

Two new phenolic amides from Allium chinense

2023/11/9 13:32:06

Objective: To isolate the phenolic amides from the dried bulbs of Allium chinense and investigate their myocardium protective activities. Methods: The chemical constituents were isolated and purified by combining with silica gel column, Sephadex LH-20 column, HPLC and other chromatography techniques. Their structures were elucidated by NMR techniques and mass spectrometry. The isolated compounds were evaluated to determine their protective effect for myocardium cells in vitro. Results: Two new phenolic amides, namely, alichinemide I (1) and alichinemide II (2), and six konwn amides were isolated from the dried bulbs of A. chinense. The structures of compounds 3–8 were identified as 3-indolcarbaldehyde (3), 1-(2-aminophenyl)urea (4), 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole3-carboxylic acid (5), N-trans-feruloyltyramine (6), N-trans-p-coumaroyltyramine (7), and N-(3,4-dimethoxyphenethyl) acetamide (8). Compound 3 (50 lmol/L) showed significant inhibitory effect on the damage of H9c2 myocardial cells induced by H2O2 in vitro. Conclusion: Compounds 1 and 2 were new phenolic amides. Compound 3 could be one of the potential myocardium protective constituents of A. chinense.

Comparative quantitative analysis of fruit oil from Hippophae rhamnoides (seabuckthorn) by qNMR, FTIR and GC–MS

2023/11/9 13:32:06

Objective: To develop a qNMR method for quantitative analysis of triacylglycerols in fruit oil of Hippophae rhamnoides (seabuckthorn, SBT) and analyze commercial samples of SBT oils using GC–MS and FTIR. Methods: SBT fruit oil (IPHRFH) was extracted with hexane and the triglyceride (TAG) was isolated by vacuum liquid chromatography. Six different branded SBT oils purchased from e-commerce suppliers (Amazon) and in-house prepared SBT oil was analyzed by qNMR and fatty acyl composition of TAGs determined by using NMR. In-house oil was also analysed by GC–MS and FTIR spectroscopy. Results: The qNMR results showed that the oil contained 80.3% of triacylglycerol (TAG). The SBT oil TAGs comprised of linolenate 6.6%, palmitoleate/oleate 65.4%, and total saturated fatty acyl chain including palmitate 28% as determined by qNMR. GC–MS analysis revealed that the major acyl functionalities present in the TAG were palmitoleic acid 36.5%, oleic acid 12.9%, palmitic acid 21.2%, and linoleic acid 18%. Of the six commercial samples analyzed, samples from only one supplier (SW) were fruit oil; All others were the seed oils or mix of fruit oil and seed oil. The labels for samples except for the SW did not indicate whether it was fruit oil or seed oil. Conclusion: The results suggest that SBT oil should be analyzed by combination of GC–MS, FTIR and qNMR for factual content of free fatty acid or TAGs, which are chemically different in nature and affect the quality of oil. GC–MS showed the content of omega free fatty acids after hydrolysis, while qNMR and FTIR showed the content of TAGs. The major acyl functionalities found in SBT fruit oil TAGs are palmitoleate/palmitate/oleate, while linoleate and linonelate make up a minor fraction. Furthermore, analysis of commercial samples showed discrepancies between label claims and actual content.

Glucuronic acid metabolites of phenolic acids target AKT-PH domain to improve glucose metabolism

2023/7/21 14:06:29

Objective: Phenolic acids widely exist in the human diet and exert beneficial effects such as improving glucose metabolism. It is not clear whether phenolic acids or their metabolites play a major role in vivo. In this study, caffeic acid (CA) and ferulic acid (FA), the two most ingested phenolic acids, and their glucuronic acid metabolites, caffeic-4'-O-glucuronide (CA4G) and ferulic-4'-O-glucuronide (FA4G), were investigated. Methods: Three insulin resistance models in vitro were established by using TNF-α, insulin and palmitic acid (PA) in HepG2 cells, respectively. We compared the effects of FA, FA4G, CA and CA4G on glucose metabolism in these models by measuring the glucose consumption levels. The potential targets and related pathways were predicted by network pharmacology. Fluorescence quenching measurement was used to analyze the binding between the compounds and the predicted target. To investigate the binding mode, molecular docking was performed. Then, we performed membrane recruitment assays of the AKT pleckstrin homology (PH) domain with the help of the PH-GFP plasmid. AKT enzymatic activity was determined to compare the effects between the metabolites with their parent compounds. Finally, the downstream signaling pathway of AKT was investigated by Western blot analysis. Results: The results showed that CA4G and FA4G were more potent than their parent compounds in increasing glucose consumption. AKT was predicted to be the key target of CA4G and FA4G by network pharmacology analysis. The fluorescence quenching test confirmed the more potent binding to AKT of the two metabolites compared to their parent compounds. The molecular docking results indicated that the carbonyl group in the glucuronic acid structure of CA4G and FA4G might bind to the PH domain of AKT at the key Arg-25 site. CA4G and FA4G inhibited the translocation of the AKT PH domain to the membrane, while increasing the activity of AKT. Western blot analysis demonstrated that the metabolites could increase the phosphorylation of AKT and downstream glycogen synthase kinase 3β in the AKT signaling pathway to increase glucose consumption. Conclusion: In conclusion, our results suggested that the metabolites of phenolic acids, which contain glucuronic acid, are the key active substances and that they activate AKT by targeting the PH domain, thus improving glucose metabolism.

An optimal medicinal and edible Chinese herbal formula attenuates particulate matter-induced lung injury through its anti-oxidative, anti-inflammatory and anti-apoptosis activities

2023/7/21 14:06:29

Objective: Identifying novel strategies to prevent particulate matter (PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases. The combined intervention represented by herbal formulae for simultaneously targeting multiple pathological processes can provide a more beneficial effect than the single intervention. The aim of this paper is therefore to design a safe and effective medicinal and edible Chinese herbs (MECHs) formula against PM-induced lung injury. Methods: PM-induced oxidative stress, inflammatory response and apoptosis A549 cell model were used to screen anti-oxidant, anti-inflammatory and anti-apoptotic MECHs, respectively. A network pharmacology method was utilized to rationally design a novel herbal formula. Ultra performance liquid chromatography-mass spectrometer was utilized to assess the quality control of MECHs formula. The excretion of magnetic iron oxide nanospheres of the MECHs formula was estimated in zebrafish. The MECH formula against PM-induced lung injury was investigated with mice experiments. Results: Five selected herbs were rationally designed to form a new MECH formula, including Citri Exocarpium Rubrum (Juhong), Lablab Semen Album (Baibiandou), Atractylodis Macrocephalae Rhizoma (Baizhu), Mori Folium (Sangye) and Polygonati Odorati Rhizoma (Yuzhu). The formula effectively promoted the magnetic iron oxide nanospheres excretion in zebrafish. The mid/high dose formula significantly prevented PM-induced lung damage in mice by enhancing the activity of SOD and GSH-Px, reducing the MDA and ROS level and attenuating the upregulation of pro-inflammatory cytokine (IL-6, IL-8, IL-1β and TNF-α), down regulating the protein expression of NF-κB, STAT3 and Caspase-3. Conclusion: Our findings suggest that the effective MECHs formula will become a novel strategy for preventing PM-induced lung injury and provide a paradigm for the development of functional foods using MECHs.

Cassiae Semen improves non-alcoholic fatty liver disease through autophagy-related pathway

2023/7/21 14:06:29

Objective: Cassiae Semen (CS, Juemingzi in Chinese) has been used for thousands of years in ancient Chinese history for relieving constipation, improving liver function as well as preventing myopia. Here we aimed to elucidate the anti-steatosis effect and underlying mechanism of CS against non-alcoholic fatty liver disease (NAFLD). Methods: High-performance liquid chromatography (HPLC) was used to identify the major components of CS water extract. Mice were fed with a high-fat and sugar-water (HFSW) diet to induce hepatic steatosis and then treated with CS. The anti-NAFLD effect was determined by measuring serum biomarkers and histopathology staining. Additionally, the effects of CS on cell viability and lipid metabolism in oleic acid and palmitic acid (OAPA)-treated HepG2 cells were measured. The expression of essential genes and proteins involved in lipid metabolism and autophagy signalings were measured to uncover the underlying mechanism. Results: Five compounds, including aurantio-obtusin, rubrofusarin gentiobioside, cassiaside C, emodin and rhein were simultaneously identified in CS extract. CS not only improved the diet-induced hepatic steatosis in vivo, as indicated by decreased number and size of lipid droplets, hepatic and serum triglycerides (TG) levels, but also markedly attenuated the OAPA-induced lipid accumulation in hepatocytes. These lipid-lowering effects induced by CS were largely dependent on the inhibition of fatty acid synthase (FASN) and the activation of autophagy-related signaling, including AMP-activated protein kinase (AMPK), light chain 3-II (LC3-II)/ LC3-1 and autophagy-related gene5 (ATG5). Conclusion: Our study suggested that CS effectively protected liver steatosis via decreasing FASN-related fatty acid synthesis and activating AMPK-mediated autophagy, which might become a promising therapeutic strategy for relieving NAFLD.

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

2023/7/21 14:06:29

Objective: The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats. Methods: Twenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment. Results: Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC. Conclusion: The present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.

Comparative chemical characters of Pseudostellaria heterophylla from geographical origins of China

Miao Sha, Xiaohuan Li, Yu Liu, Hongyue Tian, Xu Liang, Xia Li *, Wenyuan Gao — 2023/7/21 14:06:29

Objective: Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China. Methods: In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas. Results: The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value ＜ 0.05 associated with the separation of different origins were identified. Conclusion: HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.

An online automatic sorting system for defective Ginseng Radix et Rhizoma Rubra using deep learning

Qilong Xuea,b, Peiqi Miaoc, Kunhong Miaoa,b, Yang Yua,b,*, Zheng Lia,b,* — 2023/7/21 14:06:30

Objective: To establish a deep-learning architecture based on faster region-based convolutional neural networks (Faster R-CNN) algorithm for detection and sorting of red ginseng (Ginseng Radix et Rhizoma Rubra) with internal defects automatically on an online X-ray machine vision system. Methods: A Faster R-CNN based classifier was trained with around 20 000 samples with mean average precision value (mAP) of 0.95. A traditional image processing method based on feedforward neural network (FNN) obtained a bad performance with the accuracy, recall and specificity of 69.0%, 68.0%, and 70.0%, respectively. Therefore, the Faster R-CNN model was saved to evaluate the model performance on the defective red ginseng online sorting system. Results: An independent set of 2000 red ginsengs were used to validate the performance of the Faster RCNN based online sorting system in three parallel tests, achieving accuracy of 95.8%, 95.2% and 96.2%, respectively. Conclusion: The overall results indicated that the proposed Faster R-CNN based classification model has great potential for non-destructive detection of red ginseng with internal defects.

Seven new 3,4-dihydro-furanocoumarin derivatives from Angelica dahurica

2023/7/21 0:00:00

Objective: To study the chemical constituents of the roots of Angelica dahurica, a well-known Chinese herbal medicine named Baizhi in Chinese. Methods: Compounds were separated by various chromatographies, and the structures of the new compounds were elucidated based on the analysis of their spectroscopic and spectrometric data (1D, 2D NMR, HRESI MS, IR, and UV). The absolute configurations of the new compounds were determined by the calculated electronic circular dichroism and chemical derivatization. The inhibitory activities of all isolates against nitric oxide (NO) production were evaluated using lipopolysaccharide-activated RAW 264.7 macrophage cells. Results: Seven new 3,4-dihydro-furanocoumarin derivatives (1a/1b, 2a/2b, 3a/3b, 4) together with a known furanocoumarin (5) were isolated from the roots of A. dahurica. The new compounds included three pairs of enantiomers, (4S, 2′′R)-angelicadin A (1a)/(4R, 2′′S)-angelicadin A (1b), (4S, 2′′S)-angelicadin A (2a)/(4R, 2′′R)-angelicadin A (2b), and (4S, 2′′S)-secoangelicadin A (3a)/(4R, 2′′R)-secoangelicadin A (3b), together with (4R, 2′′R)-secoangelicadin A methyl ester (4). The known xanthotoxol (5) inhibited the NO production with the half-maximal inhibitory concentration (IC50) value of (32.8±0.8) μmol/L, but all the new compounds showed no inhibitory activities at the concentration of 100 μmol/L. Conclusion: This is the first report of the discovery of 3,4-dihydro-furanocoumarins from A. dahurica. The results are not only meaningful for the understanding of the chemical constituents of A. dahurica, but also enrich the reservoir of natural products.

Chemical constituents from leaves of Jatropha curcas

2023/7/21 14:06:30

Objective: To investigate the chemical constituents from the leaves of Jatropha curcas and evaluate their inhibition on lipopolysaccharide (LPS)-activated BV-2 microglia cells. Methods: The n-BuOH extract of the leaves of J. curcas was isolated by macroporous adsorption resin, silica gel, ODS, column chromatography and semi-preparative HPLC. The structures of the compounds were identified by MS, NMR, ECD, and other spectroscopic methods. In addition, anti-neuroinflammatory effects of isolated compounds were evaluated by measuring the production of nitric oxide (NO) in over-activated BV-2 cells. Results: Seventeen compounds, including (7R,8S)-crataegifin A-4-O-β-D-glucopyranoside (1), (8R,8'R)-arctigenin (2), arctigenin-4'-O-β-D-glucopyranoside (3), (-)-syringaresinol (4), syringaresinol-4'-O-β-D-glucopyranoside (5), (-)-pinoresinol (6), pinoresinol-4'-O-β-D-glucopyranoside (7), buddlenol D (8), (2R,3R)-dihydroquercetin (9), (2S,3S)-epicatechin (10), (2R,3S)-catechin (11), isovitexin (12), naringenin-7-O-β-D-glucopyranoside (13), chamaejasmin (14), neochamaejasmin B (15), isoneochamaejasmin A (16), and tomentin-5-O-β-D-glucopyranoside (17) were isolated and identified. Compounds 2, 4 and 8 significantly inhibited the release of NO in BV-2 microglia activated by LPS, with IC50 values of 18.34, 29.33 and 26.30 μmol/L, respectively. Conclusion: Compound 1 is a novel compound, and compounds 2, 3, 8, 14–17 are isolated from Jatropha genus for the first time. In addition, the lignans significantly inhibited NO release and the inhibitory activity was decreased after glycosylation.

Three new sesquiterpenes from roots of Curcuma longa

2023/7/21 14:06:30

Objective: To study the chemical constituents from the roots of Curcuma longa. Methods: The structures of the new compounds were elucidated based on extensive spectral analysis, including 1D and 2D NMR, MS, UV, and CD analysis. Results: Two new sesquiterpene compounds (1S,2R,5R,7S,8R)-2,8-epoxy-5-hydroxybisabola-3,10-dioen-9-one (1), (1R,2R,5R,7S,8R)-2,8-epoxy-5-hydroxybisabola-3,10-dioen-9-one (2), and a new natural product 6-(4-Hydroxymethylphenyl)-2-methyl-hept-2-ene-4-one (3) together with three known compounds ar-turmerone (4), 2-methyl-6-(4-hydroxyphenyl-3-methyl)-2-hepten-4-one (5) and 2-methyl-6-(4-hydroxyphenyl)-2-hepten-4-one (6) were isolated from C. longa root extract with 95% ethanol. Conclusion: In the study, three new compounds were isolated from C. longa, and their absolute configurations were determined.

Honokiol attenuates acetaminophen-induced acute liver injury by inhibiting hepatic CYP1A2 activity and improving liver mitochondrial dysfunction

2023/5/12 15:22:51

Objective: Acetaminophen (APAP) overdose is a common cause of liver injury. This study aimed to investigate the protective effect of honokiol (Hon) against APAP-induced hepatotoxicity and its potential mechanism. Methods: C57BL/6 mice were administrated with Hon (10 and 30 mg/kg) after APAP (300 mg/kg) treatment. On 1.5 h and 5 h after Hon treatment, mice were sacrificed. Serum and liver were collected. And then, liver injury-related indexes, APAP metabolism-related indexes, mitochondrial respiratory chain function-related indexes, and mitochondrial membrane function-related protein expression were evaluated. Results: It was found that Hon significantly decreased serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) activity and glutathione (GSH) depletion, increased hepatic catalase (CAT) and GSH peroxidase (GSH-Px) activities, reduced hepatic MDA and 3-nitrotyrosine contents, inhibited hepatic CYP1A2 activity and APAP protein adducts (APAP-CYS) formation. Meanwhile, oxidative phosphorylation capacity of complex I and electron transfer capacity of complex IV in mitochondrial respiratory chain was increased, whereas the release of H2O2 in the mitochondria was decreased following Hon treatment. Furthermore, Hon markedly down-regulated p-JNK in both cytosol and mitochondria, and obviously inhibited the release of apoptosis inducing factor (AIF) and endonuclease G (EndoG) from mitochondria to cytosol. Conclusion: Hon alleviated APAP-induced liver injury through the following pathways: Reducing the production of APAP-CYS by inhibiting CYP1A2 activity; Ameliorating hepatic oxidative stress by increasing the levels of hepatic CAT, GSH-Px and GSH; Improving mitochondrial respiratory chain function by promoting oxidative phosphorylation capacity of complex I and electron transfer capacity of complex IV; Improving the function of mitochondrial membrane by inhibiting p-JNK and its translocation to mitochondria, thereby reducing the release of AIF and EndoG

New insights into effects of Kaixin Powder on depression via lipid metabolism related adiponectin signaling pathway

2023/5/12 15:22:51

Objective: To clarify the anti-depressive potential mechanisms of Kaixin Powder (KP), a drug that helps to prevent and treat depression and other mentaldiseases, from genome-wide transcriptome profiling. Methods: Transcriptome and KEGG pathway analysis were conducted on the hippocampus of depressed rats, then the differentially expressed genes were validated and serum concentration of lipid parameters were identified by enzymatic assays. Furthermore, high-fat diets induced depression-like behaviors in Syrian golden hamsters were conducted to verify the predicted molecular mechanisms acquired from the transcriptome analysis. Results: Transcriptome results revealed that the 24 differentially expressed genes (DEGs) in chronic mild stress (CMS) rats could be reversed after two weeks of KP treatment. The mechanisms of KP in treating depression firstly involved the regulation of several pathology modules, including lipid metabolism, synapse function and inflammation. KP could regulate imbalances of lipid homeostasis in high-fat diet induced depressive symptoms. Furthermore, it was validated that cholesterol metabolism dysfunction can be ameliorated by KP, which was correlated with upregulation of the AdipoR1-BDNF (brain-derived neurotrophic factor) co-regulatory pathway. Conclusion: Taken together, our results demonstrated that KP not only alleviates depression via traditional mental illness targets, but it may also simulates the cholesterol metabolism and adiponectin signaling with multi-target characteristics.

Preparation of tanshinone IIA self-soluble microneedles and its inhibition on proliferation of human skin fibroblasts

2023/5/12 15:22:51

Objective: Hypertrophic scars (HS) are a variety of skin tissue fibrosis disease that occurs in human skin, the effective therapeutic method of which is still inaccessible up to now. As a bioactive constituent of a well-known medical plant, Salvia miltiorrhiza (Danshen in Chinese), tanshinone IIA (TSA) is reported to inhibit cell proliferation in HS. Therefore, the aim of this study was to prepare TSA self-soluble microneedles to strengthen its dermal retention and break through the difficulty of significantly thickening epidermal connective tissue and stratum corneum at the HS site. The possible mechanism of action in suppressing HS was studied using human skin fibroblasts (HSF). Methods: Tanshinone IIA self-dissolving microneedles (TSA-MN) was prepared using a negative mold casting method. The prescription process of microneedle was optimized by Box-Behnken effect surface method. Different media were selected to investigate the ability of transdermal absorption and in vitro release. Furthermore, according to Cell Counting Kit-8 (CCK8) method as well as the Western blot method, the effect of TSA-MN on the biological characteristics of HSF was investigated. Results: With remarkable slow release effect and dermal retention, the release and transdermal properties of TSA-MN in vitro were better than both TSA and ordinary dosage forms. Its effect of HSF confirmed the essential decrease in cell motility during cell proliferation and cell migration in vitro, which plays a significant role in down-regulating the secretion of transforming growth factor-β1(TGF-β1) in HSF and increasing the expression level of Smad7. Conclusion: The prepared TSA self-soluble microneedles is helpful in solving the problem of hypertrophic scars, with a stable dermal retention effect after process optimization.

Pharmacokinetic study about compatibility of Eucommia ulmoides and Psoralea corylifolia

2023/5/12 15:22:51

Objective: The compatibility of Eucommia ulmoides (Eu) and Psoralea corylifolia (Pc) on the pharmacokinetic (PK) properties in the rat was explored in this study. Methods: Eu extract, Pc extract and the combined extracts (crude drug ratio was 2:1) was administered by gavage, respectively. Two PK experiments were conducted. In first one, the blood samples were collected via the occuli chorioideae vein to get the PK properties of the components. In second one, the blood samples were simultaneously collected via the internal jugular vein or portal vein at different time points and the concentrations of target ingredients were detected by LC/MS/MS to clear the location where the interaction of Eu and Pc took place in vivo. Results: Eight of 11 ingredients in Eu and Pc extract were determined in rat plasma. The exposure levels of geniposidic acid (GPA), aucubin (AU), geniposide (GP), pinoresinol diglucoside (PDG), psoralen glycosides (PLG) and isopsoralen glycosides (IPLG) were decreased 1/5–2/3 after administration of combined extracts. Comparing to the combined administration, the exposure of GPA and AU in plasma of single Eu administration collected via the portal vein were decreased 1/3–2/3, and the values of AUC0-24h and AUC0-∞ of GP collected from the portal vein or internal jugular vein were double increased. The other components’ parameters were not significantly changed. Conclusion: In summary, the Pc and Eu combined administration could affect the exposure of the main components of Eu extract in rats due to the changed intestinal absorption. The research on the compatibility of Pc and Eu was helpful to guide the clinical administration of Eu and Pc simultaneously

Effect of different plant growth regulators on callus and adventitious shoots induction, polysaccharides accumulation and antioxidant activity of Rhodiola dumulosa

2023/5/12 15:22:51

Objective: As a medicinal plant, the resource of Rhodiola dumulosa is deficient along with the large collection. For the protection and utilization of R. dumulosa, the influence of plant growth regulators (PGRs) on callus induction and adventitious shoots differentiation, polysaccharide production and the antioxidant activity were tested. Methods: Internodes of R. dumulosa were used as explants and cultured on MS medium plus different plant growth regulators (PGRs). The anti-oxidative activities of polysaccharides were evaluated using radical scavenging assays. Results: By response surface plot, 0.85 mg/L N6-benzyladenine (BA), 0.34 mg/L naphthaleneacetic acid (NAA) and 0.33 mg/L 2,4-dicholorophenoxyacetic acid (2,4-D) were the optimal factors for callus induction (90.03%) from internodes explants on MS medium. The fresh weight of green callus increased 47.26 fold, when callus was inoculated on MS + thidiazuron (TDZ) 0.5 mg/L + NAA 2.0 mg/L. Adventitious buds regenerated from callus on the media of MS were fortified with BA 1.0 mg/L plus NAA 0.5 mg/L, and the induction rate was 40.00%. MS plus indole-3-butyric acid (IBA) 1.0 mg/L produced the highest rooting rate with 10 to 15 roots in a length of 2–3 cm per shoot. The content of total polysaccharides in callus developed on MS + TDZ 0.5 mg/L + NAA 2.0 mg/L and MS + BA 1.0 mg/L + NAA 0.5 mg/L was as high as 1.72%– 2.15%. At the dose of 0.5 mg/mL polysaccharides extracted from different callus induced on MS + NAA 2.0 mg/L + TDZ 0.5 mg/L or MS + BA 1.0 mg/L + NAA 0.5 mg/L or MS + BA 0.5 mg/L + 2,4-D 0.5 mg/L, the ABTS radical eliminating percentages were 82.78%, 80.18% and 68.59%, respectively, much higher than that of wild plant. Conclusion: A rapid micropropagation system for R. dumulosa has been developed. The combination of TDZ and NAA or BA and NAA can increase the yield of the total polysaccharides. The polysaccharides isolated from callus and whole wild plants had stronger free radicals scavenging activities, indicating that polysaccharides from R. dumulosa are the potential pharmaceutical supplements.

Chloroplast DNA reveals genetic population structure in Sinomenium acutum in subtropical China

2023/5/12 15:22:51

Objective: The population density and diversity of Sinomenium acutum (Menispermaceae) have been greatly reduced recently by overharvesting for medicinal purposes in China. Therefore, it is urgent that the remaining populations are investigated, and that strategies for the utilization and conservation of this species are developed. This study aimed to find the possible glacial refugia and define the genetic diversity of S. acutum for its proper utilization and conservation. Methods: A total of 77 S. acutum samples were collected from four locations, Qinling Mountains, Daba Mountains, Dalou Mountains, and Xuefeng Mountains, in subtropical China. Genetic diversity among and between these populations were phylogenetically analyzed using four chloroplast DNA molecular markers (atpI-atpH, trnQ-50 rps16, trnH-psbA and trnL-trnF). Results: A total of 14 haplotypes (C1 to C14) were found in collected samples. Haplotypes C1 and C3 were shared among all populations, with C3 as the ancestral haplotype. Haplotypes C11 and C12 diverged the most from C3 and other haplotypes. No obvious phylogeographic structure was found in four locations using the GST/NST test. There is no evidence of rapid demographic expansion in S. acutum based on the mismatch distribution, and the results of Tajima’s D test, and Fu’s FS test. Our analyses of molecular variance revealed a high level of genetic variation within populations. In contrast, the genetic differentiation among S. acutum populations was low, indicating frequent gene flow. Conclusion: Xuefeng, Dalou, and Daba Mountains were possible glacial refugia for the populations of S. acutum. C1, C3, C11 and C12 haplotypes of S. acutum should be carefully preserved and managed for their genetic value.

Analysis of codon usage patterns in Bupleurum falcatum chloroplast genome

Mengqi Gao a, Xiaowei Huo b, Liting Lu a, Mengmeng Liu a,*, Gang Zhang c,* — 2023/5/12 15:22:51

Objective: In order to distinguish the traditional Chinese medicine Bupleurum falcatum and its adulterants effectively and develop a better understanding of the factors affecting synonymous codon usage, codon usage patterns of chloroplast genome, we determine the complete chloroplast (cp) genome of B. falcatum and clarify the main factors that influence codon usage patterns of 78 genes in B. falcatum chloroplast genome. Methods: The total genomic DNA of fresh leaves from a single individual of B. falcatum was extracted with EASYspin plus Total DNA Isolation Kit and 2 lg genome DNA was sequenced using Illumina Hiseq 2500 Sequencing Platform. The cp genome of B. falcatum was reconstructed with MITObim v1.8 and annotated in the program CPGAVAS2 with default parameters. Python script and Codon W were used to calculate the codon usage bias parameters. Results: The full length of B. falcatum cp genome was 155 851 bp, 132 different genes were annotated in this cp genome containing 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. The codon usage models tended to use A/T-ending codons. The neutrality plot, ENC plot, PR2-Bias plot and correspondence analysis showed that both compositional constraint under selection and mutation could affect the codon usage models in B. falcatum cp genome. Furthermore, three optimal codons were identified and most of these three optimal codons ended with G/U. Conclusion: The cp genome of B. falcatum has been characterized and the codon usage bias in B. falcatum cp genome is influenced by natural selection, mutation pressure and nucleotide composition. The results will provide much more barcode information for species discrimination and lay a foundation for future research on codon optimization of exogenous genes, genetic engineering and molecular evolution in B. falcatum.

Cloning, expression and activity analysises of chalcone synthase genes in Carthamus tinctorius

2023/5/12 15:22:51

Objective: Flavonoids are the bioactive compounds in safflower (Carthamus tinctorius), in which chalcone synthase (CHS) is the first limiting enzyme. However, it is unclear that which chalcone synthase genes (CHSs) are participated in flavonoids biosynthesis in C. tinctorius. In this study, the CHSs in the molecular characterization and enzyme activities were investigated. Methods: Putative chalcone biosynthase genes were screened by the full-length transcriptome sequences data in C. tinctorius. Chalcone biosynthase genes in C. tinctorius (CtCHSs) were cloned from cDNA of flowers of C. tinctorius. The cloned gene sequences were analyzed by bioinformatics, and their expression patterns were analyzed by real-time PCR (RT-PCR). The protein of CtCHS in the development of flowers was detected by polyclonal antibody Western blot. A recombinant vector of CtCHS was constructed. The CtCHS recombinant protein was induced and purified to detect the enzyme reaction (catalyzing the reaction of p-coumaryl-CoA and malonyl-CoA to produce naringin chalcone). The reaction product was detected by HPLC and LC-MS. Results: Two full-length CtCHS genes were successfully cloned from the flowers of safflower (CtCHS1 and CtCHS3), with gene lengths of 1525 bp and 1358 bp, respectively. RT-PCR analysis showed that both genes were highly expressed in the flowers, but the expression of CtCHS1 was higher than that of CtCHS3 at each developmental stage of the flowers. WB analysis showed that only CtCHS1 protein could be detected at each developmental stage of the flowers. HPLC and LC-MS analyses showed that CtCHS1 could catalyze the conversion of p-coumaryl-CoA and malonyl-CoA substrates to naringin chalcone. Conclusion: CtCHS1 is involved in the biosynthesis of naringin chalcone in safflower.

Combined analysis of transcriptomics and metabolomics revealed complex metabolic genes for diterpenoids biosynthesis in different organs of Anoectochilus roxburghii

Yicong Wei a,*, Jiayuan Zhang a, Keming Qi a, Ye Li a, Ying Chen b,* — 2023/5/12 15:22:51

Objective: Diterpenoids with a wide variety of biological activities from Anoectochilus roxburghii, a precious medicinal plant, are important active components. However, due to the lack of genetic information on the metabolic process of diterpenoids in A. roxburghii, the genes involved in the molecular regulation mechanism of diterpenoid metabolism are still unclear. This study revealed the complex metabolic genes for diterpenoids biosynthesis in different organs of A. roxburghii by combining analysis of transcriptomics and metabolomics. Methods: The differences in diterpenoid accumulation in roots, stems and leaves of A. roxburghii were analyzed by metabonomic analysis, and its metabolic gene information was obtained by transcriptome sequencing. Then, the molecular mechanism of differential diterpenoid accumulation in different organs of A. roxburghii was analyzed from the perspective of gene expression patterns. Results: A total of 296 terpenoid metabolites were identified in the five terpenoid metabolic pathways in A. roxburghii. There were 38, 34, and 18 diterpenoids with different contents between roots and leaves, between leaves and stems, and between roots and stems, respectively. Twenty-nine metabolic enzyme genes with 883 unigenes in the diterpenoid synthesis process were identified, and the DXS and FDPS in the terpenoid backbone biosynthesis stage and CPA, GA20ox, GA3ox, GA2ox, and MAS in the diterpenoid biosynthesis stage were predicted to be the key metabolic enzymes for the accumulation of diterpenoids. In addition, 14 key transcription factor coding genes were predicted to be involved in the regulation of the diterpenoid biosynthesis. The expression of genes such as GA2ox, MAS, CPA, GA20ox and GA3ox might be activated by some of the 14 transcription factors. The transcription factor NTF-Y and PRE6 were predicted to be the most important transcription factors. Conclusion: This study determined 29 metabolic enzyme genes and predicted 14 transcription factors involved in the molecular regulation mechanism of diterpenoid metabolism in A. roxburghii, which provided a reference for the further study of the molecular regulation mechanism of the accumulation of diterpenoids in different organs of A. roxburghii.

De novo transcriptome assembly and metabolomic analysis of three tissue types in Cinnamomum cassia

2023/5/12 15:22:52

Objective: The barks, leaves, and branches of Cinnamomum cassia have been historically used as a traditional Chinese medicine, spice, and food preservative, in which phenylpropanoids are responsible compounds. However phenylpropanoid biosynthesis pathways are not clear in C. cassia. We elucidated the pathways by descriptive analyses of differentially expressed genes related to phenylpropanoid biosynthesis as well as to identify various phenylpropanoid metabolites. Methods: Chemical analysis, metabolome sequencing, and transcriptome sequencing were performed to investigate the molecular mechanisms underlying the difference of active components content in the barks, branches and leaves of C. cassia. Results: Metabolomic analysis revealed that small amounts of flavonoids, coumarine, and cinnamaldehyde accumulated in both leaves and branches. Transcriptome analysis showed that genes associated with phenylpropanoid and flavonoid biosynthesis were downregulated in the leaves and branches relative to the barks. The observed differences in essential oil content among the three tissues may be attributable to the differential expression of genes involved in the phenylpropanoid and flavonoid metabolic pathways. Conclusion: This study identified the key genes in the phenylpropanoid pathway controling the flavonoid, coumarine, and cinnamaldehyde contents in the barks, branches and leaves by comparing the transcriptome and metabolome. These findings may be valuable in assessing phenylpropanoid and flavonoid metabolites and identifying specific candidate genes that are related to the synthesis of phenylpropanoids and flavonoids in C. cassia.

Chemical profiling and rapid discrimination of Blumea riparia and Blumea megacephala by UPLC-Q-Exactive-MS/MS and HPLC

2023/5/12 15:22:52

Objective: To rapidly identify the two morphologies and chemical properties of similar herbal medicines, Blumea riparia and B. megacephala as the basis for chemical constituent analysis. Methods: UPLC-Q-Exactive-MS/MS was utilized for profiling and identification of the constituents in B. riparia and B. megacephala. Chemical pattern recognition (CPR) was further used to compare and distinguish the two herbs and to identify their potential characteristic markers. Then, an HPLC method was established for quality evaluation. Results: A total of 93 constituents are identified, including 54 phenolic acids, 35 flavonoids, two saccharides, one phenolic acid glycoside, and one other constituent, of which 67 were identified in B. riparia and B. megacephala for the first time. CPR indicates that B. riparia and B. megacephala samples can be distinguished from each other based on the LC–MS data. The isochlorogenic acid A to cryptochlorogenic acid peak area ratio calculated from the HPLC chromatograms was proposed as a differentiation index for distinguishing and quality control of B. riparia and B. megacephala. Conclusion: This study demonstrates significant differences between B. riparia and B. megacephala in terms of chemical composition. The results provide a rapid and simple strategy for the comparison and evaluation of the quality of B. riparia and B. megacephala.

Directional screening and identification of potential cytotoxic components from Achnatherum inebrians by a combination of surface palsmon resonance and chromatography

2023/5/12 15:22:52

Objective: To establish a method for directional screening of the cytotoxic components from the medicinal herb of Achnatherum inebrians by a combination of surface plasmon resonance (SPR) biosensor and chromatographic isolation technology. Methods: Under the guidance of bioactive assessment based on binding abilities between objects and the α-Mannosidase (α-Man) target, the active components from different solvents extracts, different polar extraction parts and fractions were screened orderly and directionally using SPR. Components with a high binding ability to a-Man can be precisely oriented in a narrower fractions range and are easy to isolate. Three human cancer cells were used to evaluate the cytotoxic activity of component with the highest affinity to α-Man. Results: Eight compounds were isolated and identificated from A. inebrians for the first time. Deoxyvasicinone possessed the highest affinity to α-Man among them. Moreover, deoxyvasicinone showed good effects on inhibited proliferation of human hepatoma cells HepG2 (IC50 = 5.7 lmol/L), human breast cancer cells MCF7 (IC50 = 7.21 lmol/L) and human lung cancer cells HCC827 (IC50 = 0.75 lmol/L), respectively. In particular, its inhibitory effect on HCC827 was stronger than the positive drug gefitinib (IC50 = 1.65 lmol/L). Conclusion: A comprehensive strategy of directional screening potential cytotoxic components from herb based on biomolecular interaction and chromatography was established. Deoxyvasicinone as an effective anti-cancer component was initially isolated from A. inebrians. It is expected that this screening strategy could provide new perspectives for rapid screening and identification of active components from natural plants with the complex matrix.

HPLC analysis of 16 compounds from Artemisia ordosica

2023/5/12 15:22:52

Objective: To establish a high-performance liquid chromatographic method (HPLC) for the simultaneous determination of 16 compounds from Artemisia ordosica. Methods: HPLC was used to analyze 16 quality indicators of A. ordosica. The HPLC conditions were as follows: Agilent Eclipse Plus C18 column (250 mm × 4.6 mm, 5 lm) with acetonitrile (A)-water (B) as mobile phase, gradient elution: 0–10 min, 75%–65% B; 10–30 min, 65%–35% B; and finally 30–40 min, 35%–15% B. The flow rate was 1.0 mL/min, the column temperature was 40 ℃, the injection volume was 10 μL, and monitored by absorbance at 285 nm for compounds 1–10, 12 and 225 nm for compounds 11, 13–16. Results: Under the selected experimental chromatographic conditions, compounds 1–16 showed good linearity (r > 0.9993) in a wide concentration range. Their average recoveries were 99.50%, 95.38%, 97.75%, 96.00%, 98.20%, 97.50%, 95.50%, 99.33%, 96.75%, 96.50%, 98.50%, 97.83%, 99.20%, 95.33%, 97.33% and 96.30%, respectively, and the RSD were 1.99%, 1.81%, 1.63%, 1.98%, 1.67%, 1.92%, 1.74%, 1.67%, 1.90%, 1.72%, 1.88%, 1.83%, 1.79%, 1.76%, 1.81% and 1.96%, respectively. Conclusion: Based on the results of the HPLC analysis, it was concluded that p-hydroxycinnamic acid (1), O-hydroxycinnamic acid (2), coniferyl alcohol (5), 5,4′-dihydroxy-7,3′-dimethoxyflavanone (8), 5,4′-dihy droxy-7-methoxyflavanone (9), 5-hydroxy-7,4′-dimethoxyflavanone (12), dehydrofalcarindiol (13), arteordoyn A (14), dehydrofalcarinol (15) and capillarin (16) are best suited for the role of quality indicators of A. ordosica grown in different ecological environments.

Cytotoxic sesquiterpene aryl esters from Armillaria gallica 012m

2023/5/12 15:22:52

Objective: To study the chemical constituents of the EtOAc extract of Armillaria gallica 012m. Methods: The chemical constituents of the EtOAc extract of A. gallica 012m were isolated and purified by various column chromatography and their structures were elucidated on the basis of the 1D and 2D NMR spectroscopic and HRESIMS data. Cytotoxicity of all isolates against A549, HCT-116, M231 and W256 human tumor cells was determined by the MTT method. Results: A new sesquiterpene aryl ester, armimelleolide C (1), and eight known ones including armillarivin (2), melleolide F (3), 6′-chloromelleolide F (4), melleolide (5), melleolide K (6), melledonol (7), 13- hydroxydihydromelleolide (8), and armillane (9), were isolated from the EtOAc extract of A. gallica 012m. All isolates showed potential cytotoxic activities against at least one of the human cancer cell lines with IC50 values ranging from (3.17 ± 0.54) to (17.57 ± 0.47) lmol/L. Compound 1 showed significant inhibitory activity against M231 with an IC50 value of (7.54 ± 0.24) lmol/L compared with paclitaxel as the positive control. Compounds 2, 3, and 7, 9 showed obvious inhibitory activity against HCT-116 and were better than that of the positive control. Conclusion: The chemical constituents including a new sesquiterpene aryl ester armimelleolide C (1) from the EtOAc extract of A. gallica 012m have a variety of structures and potential antitumor activities.

Temporal characteristics of agarwood formation in Aquilaria sinensis after applying whole-tree agarwood-inducing technique

2023/1/30 8:18:45

Objective: Agarwood—a resinous wood produced by Aquilaria plants in response to injury or artificial induction—is a valuable medicinal and fragrance resource. Whole-Tree Agarwood-Inducing Technique (Agar-WIT) has been widely used to produce agarwood. However, the time-dependent characteristics of agarwood formation induced by Agar-WIT are yet to be clarified. To promote technologically efficient utilization and upgradation of Agar-WIT, the dynamic process and mechanism of agarwood formation were analyzed for one year. Methods: Agarwood formation percentage, barrier layer microscopic properties, extract levels, compound level, and characteristic chromatograms of agarwood were examined by referring to the Chinese Pharmacopeia (2020 version). Results: Agar-WIT could maintain a high percentage of agarwood formation over one year compared with that of healthy plants. Alcohol-soluble extract and agarotetrol levels showed fluctuating cyclic changes with peaks occurring first during the fifth and sixth months, and subsequently in the 11th month. Aquilaria trees subjected to Agar-WIT treatment for 1–12 months showed significant characteristics of a dynamic agarwood formation process. The barrier layer began to appear in the fourth month after treatment. Alcohol-soluble extractive levels in agarwood formed in the second month, and thereafter, exceeded 10.0%, and agarotetrol in agarwood produced after four months or later, exceeded 0.10%. Conclusion: According to the Chinese Pharmacopoeia, alcohol-soluble extractive levels in agarwood should not be less than 10.0% and agarotetrol level should exceed 0.10%. After four months of Agar-WIT treatment, the formed agarwood theoretically met these standards and was suitable for developed and utilization. However, the optimal harvest time was found to be the 11th month, followed by the sixth month after Agar-WIT treatment. Therefore, Agar-WIT resulted in swift agarwood formation and stable accumulation of alcohol-soluble extracts and agarotetrol. Thus, this method is efficient for large-scale cultivation of Aquilaria sinensis to produce agarwood and provide raw materials for the agarwood medicinal industry.

Bacillus and microalgae biofertilizers improved quality and biomass of Salvia miltiorrhiza by altering microbial communities

Xuemin Wei 1, Xuanjiao Bai 1, Pei Cao, Gang Wang, Jianping Han *, Zheng Zhang * — 2023/1/30 8:18:45

Objective: Biofertilizers are reliable alternatives to chemical fertilizers due to various advantages. However, the effect of biofertilizers on Salvia miltiorrhiza yield and quality and the possible mechanisms remain little known. Here, an experiment was conducted in S. miltiorrhiza field treated with two kinds of biofertilizers including Bacillus and microalgae. Methods: A field experiment was conducted on S. miltiorrhiza of one year old. The biofertilizers were applied at six treatments: (i) control check, CK; (ii) microalgae, VZ; (iii) Bacillus, TTB; (iv) microalgae + Bacillus (1:1), VTA; (v) microalgae + Bacillus (0.5:1), VTB; (vi) microalgae + Bacillus (1:0.5), VTC. Here, high-throughput sequencing, ICP-MS and UPLC were employed to systematically characterize changes of microbial diversity and structure composition, heavy metals content and bioactive compounds, respectively. Results: Compared to CK, root biomass increased by 29.31%-60.39% (P < 0.001). Meanwhile, bioactive compounds were higher than CK after the application of the biofertilizers, peculiarly in TTB and VTB. However, the content of Pb contents in roots significantly reduced by 46.03% and 37.58% respectively in VTC and TTB (P < 0.05). VTA application notably increased the available nitrogen content by 53.03% (P < 0.05), indicating the improvement of soil fertility. Significantly, bacterial and fungal Chao I diversity indices showed an increasing trend with biofertilizer application (P < 0.05), and biofertilizer amendment enriched the rhizosphere soil with beneficial microorganisms that have abilities on promoting plant growth (Achromobacter and Penicillium), adsorbing heavy metal (Achromobacter and Beauveria), controlling plant pathogen (Plectosphaerella, Lechevalieria, Sorangium, Phlebiopsis and Beauveria) and promoting the accumulation of metabolites (Beauveria and Phoma). Conclusion: Bacillus and microalgae biofertilizers improved the quality and biomass of S. miltiorrhiza by altering microbial communities in soil.

Auxin induces lateral root formation in Bupleurum: A heme oxygenase dependent approach

2023/1/30 8:18:45

Objective: The content of saikosaponins in genus Bupleurum is increased with numbers of lateral root, but the genetic mechanisms are largely unknown. This study aims to identify the heme oxygenase (HO) gene family members of B. chinense and B. scorzonerifolium, and assess their role in the root development in Bupleurum. Methods: The gene sequences of HO family were selected from iso-seq full-length transcriptome data of B. chinense and B. scorzonerifolium, and were analyzed in physicochemical properties, conserved domains, motifs and phylogenetic relationship. In addition, the expression patterns of HO gene in different parts of roots were compared via transcriptome sequencing and qRT-PCR in the two species. Results: Five Bupleurum HO genes (BcHO1-BcHO5) belonging to the HO1 subfamily were identified from the transcriptome data, whereas the HO2 subfamily member was not identified. The expression levels of BcHO1 and BcHO2 were significantly higher than those of other three HO members in the transcriptome analysis. In addition, the expression profile of BcHO1 showed consistency with lateral root development in B. chinense and B. scorzonerifolium. Conclusion: Hos might participate in the auxin-induced morphogenesis of lateral roots. The yield of saikosaponin may be improved by manipulating expression of these genes.

Geographical discrimination of Cyclocarya paliurus tea for origin traceability based on multielement analysis by ICP-OES and chemometrics multivariate

Na Guo, Qi Wu, Chen Shi, Rengeng Shu * — 2023/1/30 8:18:46

Objective: This paper focused on the geographical discrimination of Cyclocarya paliurus tea for origin traceability based on multielement analysis by ICP-OES and chemometrics multivariate. Methods: In this study, eleven trace element concentrations were determined by ICP-OES and processed by multivariate statistical analysis. Results: Based on ANOVA, the mean concentrations of 10 elements except Co differed significantly among six origins. Pearson’s correlation analysis showed that 11 pairs of elements have a positive significant correlation and 12 pairs have a negative significant correlation. The geographical origins were effectively differentiated using the eleven elements combined with PCA. And the S-LDA model offered a 100% differentiation rate. Conclusion: The overall results suggested that the combination of multielement analysis by ICP-OES and chemometrics multivariate could trace the geographical origins of tea. And the paper can provide reference for quality control and quality evaluation of C. paliurus in the future.

Identification of microRNA and analysis of target genes in Panax ginseng

2023/1/30 8:18:46

Objective: Ginsenosides, polysaccharides and phenols, the main active ingredients in Panax ginseng, are not different significantly in content between 3 and 5 years old of ginsengs called Yuan ginseng and more than ten years old ones called Shizhu ginseng. The responsible chemical compounds cannot fully explain difference in efficacy between them. According to reports in Lonicerae Japonicae Flos (Jinyinhua in Chinese) and Glycyrrhizae Radix et Rhizoma (Gancao in Chinese), microRNA may play a role in efficacy, so we identified microRNAs in P. ginseng at the different growth years and analyzed their target genes. Methods: Using high-throughput sequencing, the RNA-Seq, small RNA-Seq and degradome databases of P. ginseng were constructed. The differentially expressed microRNAs was identified by qRT-PCR. Results: A total of 63,875 unigenes and 24,154,579 small RNA clean reads were obtained from the roots of P. ginseng. From these small RNAs, 71 miRNA families were identified by bioinformatics target prediction software, including 34 conserved miRNAs, 37 non-conserved miRNA families, as well as 179 target genes of 17 known miRNAs. Through degradome sequencing and computation, we finally verified 13 targets of eight miRNAs involved in transcription, energy metabolism, biological stress and disease resistance, suggesting the significance of miRNAs in the development of P. ginseng. Consistently, major miRNA targets exhibited tissue specificity and complexity in expression patterns. Conclusion: Differential expression microRNAs were found in different growth years of ginsengs (Shizhu ginseng and Yuan ginseng), and the regulatory roles and functional annotations of miRNA targets in P. ginseng need further investigation.

Lung protective effects of dietary malate esters derivatives from Bletilla striata against SiO2 nanoparticles through activation of Nrf2 pathway

Di Zhou, Wenhui Chang, Jiaxin Qi, Gang Chen, Ning Li * — 2023/1/30 8:18:47

Objective: To study the protective activities of the dietary malate esters derivatives of Bletilla striata against SiO2 nanoparticles-induced A549 cell lines and its mechanism action. Methods: The components were isolated and elucidated by spectroscopic methods such as 1D NMR and 2D NMR. And MTT assays was used to tested these components on the A549 cell survival rates and ROS or proteins levels were detected by Western blotting. Results: A new glucosyloxybenzyl 2-isobutylmalate (a malate ester derivative), along with 31 known compounds were isolated and identified from n-BuOH extract of EtOH extract of B. striata. Among them, compounds 3, 4, 11, 12 and 13 possessed noteworthy proliferative effects for damaged cells, with ED50 of 14.0, 13.1, 3.7, 11.6 and 11.5 lmol/L, respectively, compared to positive control resveratrol (ED50, 14.7 lmol/L). Militarine (8) prominently inhibited the intracellular ROS level, and increased the expression of Nrf2 and its downstream genes (HO-1 and c-GCSc). Furthermore, Nrf2 activation mediates the interventional effects of compound 8 against SiO2 nanoparticles (nm SiO2)-induced lung injury. Moreover, treatment with compound 8 significantly reduced lung inflammation and oxidative stress in nm SiO2-instilled mice. Molecular docking experiment suggested that 8 bound stably to the HO-1 protein by hydrogen bond interactions. Conclusion: The dietary malate esters derivatives of B. striata could significantly increase the viability of nm SiO2-induced A549 cells and decrease the finer particles-induced cell damages. Militarine is especially promising compound for chemoprevention of lung cancer induced by nm SiO2 through activation of Nrf2 pathway.

Protective effect of hydroxysafflor yellow A on MSCs against senescence induced by D-galactose

Xiaoqing Song *, Jinying Wang, Yu Zhang, Xinqian Du, Qibing Qian — 2023/1/30 8:18:48

Objective: To examine the protective effects of hydroxysafflor yellow A (HSYA) against the senescence of mesenchymal stem cells (MSCs) induced by D-galactose (D-gal) in vitro, and investigate the potential mechanism involved. Methods: Grouping experiment, Normal control (NC) group: conventional culture with complete medium; Senescence group: MSCs were cultured for 48 h with complete medium containing 10 g/L D-gal; HSYA group: on the basis of senescence induction, HSYA with the suitable concentration was used to protect MSCs. The key experimental indices associated with oxidative stress, inflammatory response, cell senescence, proliferation and apoptosis were measured through chemical colorimetry, b-galactosidase staining, EdU incorporation and flow cytometry, respectively. The relative quantity (RQ) of proteins related closely to cell proliferation, apoptosis, and NF-jB signaling were measured by Western blotting. Results: As compared with Senescence group, treatment with HSYA (120 mg/L) effectively ameliorated the adverse situation of MSCs. Oxidation stress and inflammation along with D-Gal induction was dramatically alleviated in MSCs; The b-Gal-positive staining indicated that MSC senescence was significantly mitigated; The proliferative capability of MSCs was significantly increased by up-regulating PCNA and inhibiting p16 expression; The anti-apoptotic effect on MSCs was exerted by down-regulating the RQ of cleaved Caspase-3 and Bax; The activity of NF-jB signaling in MSCs was notably suppressed through inhibiting phosphorylation of IKKb and p65. Conclusion: HSYA (120 mg/L) significantly delayed the D-Gal-induced senescence process in MSCs through attenuating inflammatory reaction and oxidative stress, and suppressing the activity of NF-jB signaling.

Pterostilbene supresses inflammation-induced melanoma metastasis by impeding neutrophil elastase-mediated thrombospondin-1 degradation

2023/1/30 8:18:48

Objective: Chronic inflammation plays a fatal role in tumor metastasis. Pterostilbene (PTE) is a natural dimethylated analogue of resveratrol with anticancer and anti-inflammatory activities. This study aimed to investigate the inhibitory effect of PTE on inflammation-associated metastasis and explore the underlying mechanisms. Methods: Lipopolysaccharide (LPS)-induced lung inflammation and melanoma metastasis models were established in mice. After PTE treatment for four weeks, the organ index, histological changes, proinflammatory cytokines, and the expression and activity of neutrophil elastase (NE), a biomarker of neutrophil influx in the lungs, were analysed. Additionally, direct effects of PTE on NE-induced B16 cell migration were explored in wound healing and Transwell assays, and the expression of thrombospondin-1 (TSP-1) and epithelial-mesenchymal transition (EMT) markers were also detected. Results: PTE obviously attenuated the LPS-induced metastasis of circulatory B16 cells to lungs by reducing the number of metastatic nodules on the lung surfaces and the lung weight/body weight ratio. PTE treatment also significantly reduced LPS-activated increase levels of tumor necrosis factor (TNF)-a and interleukin (IL)-6 in the lungs of tumor-bearing mice. In addition, increased expression and enzyme activity of NE and decreased expression of TSP-1 were observed, and these were blocked by PTE. In vitro, PTE at concentrations without cytotoxicity also markedly suppressed NE-triggered B16 cell migration, prevented NE-induced TSP-1 proteolysis and reversed the expression of vimentin, N-cadherin and Ecadherin. Conclusion: PTE could block inflammation-enhanced tumor metastasis, and the underlying mechanism might be associated with the inhibition of NE-mediated TSP-1 degradation.

Protective effect of Cistanche deserticola on gentamicin-induced nephrotoxicity in rats

2023/1/30 8:18:48

Objective: Gentamicin (GM) is a commonly used aminoglycoside antibiotic, however, renal toxicity has limited its usage. The present study was designed to evaluate the ameliorative effect of Cistanche deserticola on GM-induced nephrotoxicity in rats. Methods: The nephrotoxicity in rats was induced by intraperitoneal administration of GM (100 mg/kg) for 10 consecutive days. Glomerular filtration rate, blood urea nitrogen, creatinine and kidney histopathology were detected to assess the GM-induced nephrotoxicity. The oxidative stress (catalase, superoxide dismutase, glutathione and malondialdehyde) was assessed. The inflammatory response (tumor necrosis factor-a, interleukin-6, myeloperoxidase and nuclear factor-kappa B) and apoptotic marker (Bax and Bcl-2) were also evaluated. Results: The results showed that water and 75% ethanol extracts of C. deserticola (named CDW and CDE, respectively) (100, 200 and 400 mg/kg) in combination with GM could recover the reduction of glomerular filtration rate and enhance the renal endogenous antioxidant capability induced by GM. The increase in the expression of renal inflammatory cytokines (tumor necrosis factor-a and interleukin-6), nuclear protein of nuclear factor-kappa B (p65) and the activity of myeloperoxidase induced by GM was significantly decreased upon CDW or CDE treatment. In addition, CDW or CDE treatment could decrease the Bax protein expression and increase the Bcl-2 protein expression in GM-induced nephrotoxicity in rats significantly. Conclusion: The study demonstrated that C. deserticola treatment could attenuate kidney dysfunction and structural damage in rats induced by GM through the reduction of inflammation, oxidative stress and apoptosis.

Comparison of intestinal absorption characteristics between rhubarb traditional Chinese medicine preparation and activity ingredients using in situ and in vitro model

2023/1/30 8:18:48

Objective: The intestinal absorption characteristics of active ingredients are very important for oral administration of traditional Chinese medicine (TCM) to achieve the desired therapeutic effect. However, a deeper understanding about active ingredients absorption characteristics is still lack. The aim of this study was to investigate the absorption properties and mechanism of rhubarb active ingredients in TCM preparation and pure form. Methods: The intestinal absorption behavior of active ingredients in Shenkang extract (SKE) and rhubarb anthraquinone ingredients (RAI) were investigated by in situ single-pass intestinal perfusion model. And the bidirectional transport characteristics of these active ingredients were assessed by in vitro Caco-2 cell monolayer model. Results: In situ experiment on Sprague-Dawley rats, the effective permeability coefficient values of aloeemodin, emodin and chrysophanol in RAI were higher than those in SKE, and the value of rhein in RAI was lower than that in SKE. But the easily absorbed segments of intestine were consistent for all ingredients, whether in SKE or in RAI. In vitro experiment, the apparent permeability coefficient values of rhein, emodin and chrysophanol in RAI were higher than those in SKE, and this value of aloe-emodin in RAI was lower than that in SKE. But their efflux ratio (ER) values in SKE and RAI were all similar. Conclusion: Four rhubarb anthraquinone ingredients in SKE and RAI have similar absorption mechanism and different absorption behavior, and the microenvironment of the study models influenced their absorption behavior. The results may provide an aid for understanding of the absorption characteristics of the TCM active ingredients in complex environments and the complementarities of different research models.

α-Ionone protects against UVB-induced photoaging in epidermal keratinocytes

Ruixuan Geng a, Seong-Gook Kang b, Kunlun Huang a,c,d, Tao Tong a,c,d,* — 2023/1/30 8:18:49

Objective: To evaluate whether α-ionone, an aromatic compound mainly found in raspberries, carrots, roasted almonds, fruits, and herbs, inhibits UVB-mediated photoaging and barrier dysfunction in a human epidermal keratinocyte cell line (HaCaT cells). Methods: The anti-photoaging effect of α-ionone was evaluated by detecting the expression of barrier-related genes and matrix metalloproteinases (MMPs) in HaCaT cells. The levels of reactive oxygen species, oxidation product, antioxidant enzyme, and inflammatory factors were further analysed to underline the protective effect of α-ionone on epidermal photoaging. Results: It was found that α-ionone attenuated UVB-induced barrier dysfunction by reversing keratin 1 and filaggrin in HaCaT cells. α-Ionone also reduced the protein amount of MMP-1 and mRNA expression of MMP-1 and MMP-3 in UVB-irradiated HaCaT cells, implying protective effects on extracellular matrix. Furthermore, HaCaT cells exposed to a-ionone showed significant decreases in interleukin (IL)-1b, IL-6, IL-8, and tumor necrosis factor-a as compared to UVB-irradiated HaCaT cells. a-Ionone treatment significantly inhibited the UVB-induced intracellular reactive oxygen species increase and malondialdehyde accumulation. Therefore, the beneficial effects of a-ionone on inhibiting MMPs secretion and barrier damage may be related to attenuated inflammation and oxidative stress. Conclusion: Our results highlight the protective effects of α-ionone on epidermal photoaging and promote its clinic application as a potential natural anti-photodamage agent in future.

Identification of prototype compounds and their metabolites in rats’ serum from Xuefu Zhuyu Decoction by UPLC-Q-TOF/MS

2023/1/30 8:18:49

Objective: As a classic prescription in traditional Chinese medicine, Xuefu Zhuyu Decoction (XFZYD) has been widely used in the clinical treatment of cardiovascular and cerebrovascular diseases. In order to unveil the potentially effective compounds, a rapid ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) method was established to identify prototype compounds and their metabolites from XFZYD in rats’ serum. Methods: The serum from rats after intragastric administration of XFZYD aqueous extract was analyzed by UPLC-Q-TOF/MS method. The prototype compounds and their metabolites were identified by comparison with the reference standards and tentatively characterized by comprehensively analyzing the retention time, MS data, characteristic MS fragmentation pattern and retrieving literatures. Results: A total of 175 compounds (24 prototype compounds and 151 metabolites) were identified and tentatively characterized. The metabolic pathways of prototype compounds in vivo were also summarized, including glucuronidation, hydrolyzation, sulfation, demethylation, and hydroxylation, and so on. Conclusion: In this study, a UPLC-Q-TOF/MS technique was developed to analyze prototype compounds and their metabolites from XFZYD in serum, which would provide the evidence for further studying the effective compounds of XFZYD.

Chemical constituents from aerial parts of Scoparia dulcis

2023/1/30 8:18:49

Objective: To study the chemical constituents from the aerial parts of Scoparia dulcis. Methods: Various chromatographic techniques were used to separate the constituents and their structures were elucidated using spectroscopic methods and by comparing their data to those reported in the literatures. The α-glucosidase inhibitory activity assay was used to identify potential α-glucosidase inhibitors. Results: Nine compounds were isolated from the aerial parts of S. dulcis. Their structures were identified as Scoparic zolone (1), (2S)-2,7-dihydroxy-2H-1,4-benzoxazin-3(4H)-one (2), (2R)-7-hydroxy-2H-1,4- benzoxazin-3(4H)-one-2-O-β-D-glucopyranoside (3), (2R)-7-methoxy-2H-1,4-benzoxazin-3(4H)-one-2- O-β-D-glucopyranoside (4), (2S)-7-hydroxy-2H-1,4-benzoxazin-3(4H)-one-2-O-β-D-glucopyranoside (5), 6-methoxy-benzoxazolin-2(3H)-one (6), 4-acetonyl-3,5-dimethoxy-p-quinol (7), zizyvoside I (8), and 3,4-dihydroxy benzeneacetic acid (9). Compound 2 showed the potent α-glucosidase inhibitory activity with an IC50 value of (132.8 ± 11.5) lmol/L, which is 28-fold higher than the positive control acarbose. Conclusion: Compound 1 is a new natural product. Compounds 2 and 9 have not been reported in Scoparia before. Compounds 3, 5, 7, 8 are isolated from Scrophulariaceae for the first time.

Phylogenetic and spatial patterns of herbal medicine compounds: Which medicinal plants are phytochemically characterized?

Dacheng Hao a,1,*, Yaoxuan Wang a,1, Peigen Xiao b, Xiaojie Gu a,* — 2024/11/13 14:19:56

Objective: The study of phytometabolites of medicinal plants and their phylogenetic distribution is an important content of pharmacophylogeny. The objectives of this study were to provide an updated estimate of the extent to which the medicinal plants were investigated phytochemically and relate this to the species-level phylogeny and their geographical pattern. Methods: Here, we further characterized 1 648 phytometabolites, including terpenoids, steroids, flavonoids, phenylpropanoids, phenolics, alkaloids, etc., reported in journals Chinese Traditional and Herbal Drugs (Zhong Cao Yao) and Chinese Herbal Medicines (CHM) from the phylogenetic and spatial perspectives. According to the structural characteristics, terpenoids, flavonoids, alkaloids and phenylpropanoids were subdivided into subclasses, and the research effort of phytometabolites was for the first time delineated at both subclass and phylogenetic levels. Results: The phytometabolites of 90 families were reported on two journals in three years. It is noted that terpenoids with diverse bioactivities still constitute the primary object of phytochemical research, followed by flavonoids, phenolics, phenylpropanoids and alkaloids. Among the reported species, the family Asteraceae had the most, followed by Lamiaceae, Fabaceae, and Ranunculaceae. In the phylogenetic distribution of the reported phytometabolites, the net relatedness index (NRI) results revealed a clustered structure for triterpene, iridoid, flavone, flavonol, coumarin, indole alkaloid and terpenoid alkaloid, while the nearest taxon index (NTI) metric identified the clustered structure for triterpene, sesquiterpene, indole alkaloid and terpenoid alkaloid. Especially in Ranunculaceae, there were more reports on triterpene and terpenoid alkaloid subclasses. The overdispersion pattern of diterpene and phenolic was suggested by NRI and NTI respectively, albeit more reported diterpenes and phenolics were in Lamiaceae. The geographical distribution hotspots of reported species and compounds thereof highlighted the enormous progress of herbal medicine research and industry, which play a positive role in the future drug discovery and development. Conclusion: These results provide new dimensions and perspectives in the context of pharmacophylogeny for perceiving and evaluating research trends and flashpoints in medicinal phytochemistry.

Metabolomics combined with network pharmacology reveals anti-asthmatic effects of Nepeta bracteata on allergic asthma rats

2024/11/13 14:19:56

Objective: To investigate the mechanisms that underlie the anti-asthmatic effects of Nepeta bracteata (DBJJ, Dabao Jingjie in Chinese) in rats by integrating metabolomics and network pharmacology. Methods: In this study, the rat model of asthma was induced by ovalbumin (OVA), and the rats were treated with a decoction of N. bracteata. Pathological changes in lung tissue were observed, and the quantification of eosinophils (EOS) and white blood cells (WBC) in bronchoalveolar lavage fluid was performed. Furthermore, the serum levels of asthma-related factors induced by OVA were assessed. 1H NMR spectroscopy serum metabolomics method was utilized to identify differential metabolites and their associated metabolic pathways. UPLC-QE-MS/MS combined with network pharmacology was employed to predict the core targets and pathways of DBJJ in its action against asthma. The anti-asthmatic properties of DBJJ were investigated using an integrated approach of metabolomics and network pharmacology. The findings were validated through molecular docking and Western blotting analysis of the key targets. Results: The administration of DBJJ effectively alleviated OVA-induced lung histopathological changes and decreased the number of EOS and WBC in BALF. Additionally, DBJJ inhibited the OVA-induced elevation of TNF-a, IL-18, Ig-E, EOS, IL-1b, MDA, VEGF-A, and TGF-b1. A total of 21 biomarkers and 10 pathways were found by metabolomics analysis. A total of 29 compounds were identified by UPLC-QE-MS/MS, in which 13 active components were screened by oral availability and Caco-2 cell permeability, the 120 targets and 173 KEGG pathways were predicted. The integration of metabolomics and network pharmacological analysis revealed that DBJJ’s main constituents, including ferulic acid and ursolic acid, exerted their effects on four targets, namely DAO and NOS2, as well as their associated metabolites and pathways. The active constituents of DBJJ demonstrated a high binding affinity towards DAO and NOS2. Furthermore, DBJJ was observed to decrease the protein expression and phosphorylation levels of NOS2, MAPK, and STAT3. Conclusion: The administration of DBJJ demonstrates notable anti-asthma properties in rats with allergic asthma. This effect can be attributed to the modulation of various targets, including NOS2, MAPK, and STAT3, by primary constituents such as ferulic acid and ursolic acid.

Deciphering relationship between depression and microbial molecules based on multi-omics: A case study of Chaigui Granules

Qi Wang a,b, Yingxia Zhao a,b, Xuemei Qin a,b,*, Junsheng Tian a,b,* — 2024/11/13 14:19:56

Objective: To decipher the antidepression effect of Chaigui Granules (CGKL) from the relationship between depression and microbial molecules based on multi-omics. Methods: Male SD rats were subjected to chronic unpredictable mild stress (CUMS) for seven weeks. The antidepressants CGKL extract and CGKL were administered for the following four weeks. The behavior test and the content of monoamine neurotransmitters were used to evaluate the efficacy of CGKL. The 16S rRNA sequencing, LC-MS technology and molecular biological techniques were used to explore the pharmacological mechanism of CGKL. Results: CGKL treatment obviously alleviated the depressive behavioral indicators and regulated the content of monoamine neurotransmitters, and presented dose-dependent manner. CGKL could also improve the arginine metabolism disorder of gut microbiota in the jejunum. Meanwhile, the contents of arginine and its metabolites in the serum and hippocampus were regulated to normal levels. Further investigation indicated that the expression of related rate-limiting enzyme genes and proteins in the hippocampus was validated by qRT-PCR and Western blotting. The results showed that the gut microbiota, metabolites, and genes or proteins of rate-limiting enzymes involved in the arginine pathway were significantly regulated by CGKL. Conclusion: The present study demonstrates that CGKL might exert antidepressant effects through regulating arginine metabolism, and its mechanism may be related to modulating the gut microbiota and related metabolic enzyme.

Compound Danshen Dripping Pills combined with isosorbide mononitrate for angina pectoris: A systematic review and a Meta-analysis

Ru Wang, Jing Hu, Yuanyuan Li, Hong Yin * — 2024/11/13 14:19:56

Objective: To evaluate the efficacy of Compound Danshen Dripping Pills (CDDP) combined with isosorbide mononitrate (ISMN) versus ISMN alone for treating angina pectoris in patients. Methods: The PubMed, Web of Science, Cochrane Library, Embase China National Knowledge Infrastructure, China Biomedical Literature Service System, Chinese Medical Journal Database, and Wan Fang MED databases were searched from inception to November 2022. Randomized controlled trials (RCTs) and cohort studies were included. The primary outcomes were angina symptom and electrocardiography (ECG) efficacy, angina symptom efficacy, and ECG efficacy. The protocol was registered with PROSPERO No. CRD42022314774. Results: Our study included 7 245 patients with angina (59 RCTs, 11 cohort studies). When ISMN was combined with CDDP, the efficacy of angina symptom and ECG [odds ratio (OR) = 4.824, 95% confidence interval (CI) = 3.636–6.401, P = 0.000], the efficacy of angina symptom (OR = 4.347, 95% CI = 3.635–5.198, P = 0.000), the efficacy of ECG (OR = 3.364, 95% CI = 2.767–4.089, P = 0.000) were better than that of patients treated with ISMN alone. CDDP combined with ISMN was superior to ISMN alone in reducing triglyceride (TG) [mean difference (MD) =–35.176, 95% CI = –37.439 to–32.912, P = 0.000], total cholesterol (TC) (MD = –24.296, 95% CI =–26.429 to –22.163, P = 0.000), the duration of angina attack (MD =–1.991, 95% CI =–2.349 to–1.633, P = 0.000), and the frequency of angina attack [standardized MD (SMD) = –2.840, 95% CI =–3.416 to–2.265, P = 0.000]. There was no increase in adverse events between CDDP combined with ISMN and ISMN alone (OR = 0.513, 95% CI = 0.421–0.626, P = 0.000). Conclusion: CDDP combined with ISMN improved treatment efficacy and was well tolerated. Therefore, this combination could be used as an alternative treatment. However, clinical and patient conditions should be considered.

Network pharmacology, molecular docking, and untargeted metabolomics reveal molecular mechanisms of multi-targets effects of Qingfei Tongluo Plaster improving respiratory syncytial virus pneumonia

Mengfei Yang a, Xiuying Zhang b,*, Qing Liu a, Yongxue Wang a — 2024/11/13 14:19:56

Objective: Qingfei Tongluo Plaster (QFP), an improved Chinese medicine hospital preparation, is an attractive treatment option due to its well clinical efficacy, convenience, economy, and patient compliance in the treatment of respiratory syncytial virus (RSV) pneumonia. The aim of this study was to investigate the efficacy mechanism of QFP on RSV rats from the perspective of alleviating lung inflammation and further explore the changes of serum metabolites and metabolic pathways in RSV rats under the influence of QFP. Methods: This study used network pharmacological methods and molecular docking combined with molecular biology and metabolomics from multi-dimensional perspectives to screen and verify the therapeutic targets. Open online databases were used to speculate the gene targets of efficient ingredients and diseases. Then, we used the String database to examine the fundamental interaction of common targets of drugs and diseases. An online enrichment analysis was performed to predict the functional pathways. Molecular docking was applied to discover the binding modes between essential ingredients and crucial gene targets. Finally, we demonstrated the anti-inflammatory ability of QFP in the RSV-evoked pneumonia rat model and explained the mechanism in combination with the metabolomics results. Results: There were 19 critical targets defined as the core targets: tumor necrosis factor (TNF), inducible nitric oxide synthase 2 (NOS2), mitogen-activated protein kinase 14 (MAPK14), g1/S-specific cyclin-D1 (CCND1), signal transducer and activator of transcription 1 alpha/beta (STAT1), proto-oncogene tyrosine-protein kinase Src (SRC), cellular tumor antigen p53 (TP53), interleukin-6 (IL6), hypoxiainducible factor 1-alpha (HIF1A), RAC-alpha serine/threonine-protein kinase (AKT1), signal transducer and activator of transcription 3 (STAT3), heat shock protein HSP 90-alpha (HSP90AA1), tyrosine-protein kinase JAK2 (JAK2), cyclin-dependent kinase inhibitor 1 (CDKN1A), mitogen-activated protein kinase 3 (MAPK3), epidermal growth factor receptor (EGFR), myc proto-oncogene protein (MYC), protein c-Fos (FOS) and transcription factor p65 (RELA). QFP treated RSV pneumonia mainly through the phosphatidylinositol 3-kinase (PI3K)/RAC AKT pathway, HIF-1 pathway, IL-17 pathway, TNF pathway, and MAPK pathway. Animal experiments proved that QFP could effectively ameliorate RSV-induced pulmonary inflammation. A total of 28 metabolites underwent significant changes in the QFP treatment, and there are four metabolic pathways consistent with the KEGG pathway analyzed by network pharmacology, suggesting that they may be critical processes related to treatment. Conclusion: These results provide essential perspicacity into the mechanisms of action of QFP as a promising anti-RSV drug.

Mechanisms of Shufeng Jiedu Capsule in treating bacterial pneumonia based on network pharmacology and experimental verification

2024/11/13 14:19:56

Objective: The aim of this study was to investigate the underlying mechanism of Shufeng Jiedu Capsule (SFJD) for treating bacterial pneumonia (BP) in vivo based on network pharmacology and experimental verification study. Methods: Network pharmacology was used to screen the active compounds and target genes of SFJD. Then, the multi drug resistance-Pseudomonas aeruginosa (MDR-PA) mice lethal model and MDR-PA pneumonia model were established to evaluate the therapeutic effects and underlying mechanisms of SFJD. Western blot and ELISA were used to determinate the protein expression level of the IL-17 signaling pathway and JAK/STAT signaling pathway. Results: After screening, 172 potential components of SFJD were generated, based on which we constructed an SFJD-component-target-BP interaction network. The Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment revealed that SFJD could regulate the IL-17 signaling pathway and Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Molecular docking showed that the potential target proteins had good combinations with the main active components. SFJD significantly reduced the mortality and prolonged survival days in lethal models. The lung index and pathological changes in the lung were also significantly decreased. SFJD could significantly decrease the expression of interleukin-17A (IL-17A), TNF receptor associated factor 6 (TRAF6), phospho-inhibitor of nuclear factor-kappa B (p-IjB)/inhibitor of NF-jB (IjB), phospho-NF-jB p65 (p-NF-jB p65), phospho-protein kinase B (p-AKT)/AKT, phospho-signal transducer and activator of transcription 3 (p -STAT3)/STAT3, phospho-signal transducer and activator of transcription 1 (p-STAT1)/STAT1, and the protein level of interleukin-6 (IL-6), tumor necrosis factor a (TNF-a), and IL-1b. Conclusion: Combined with network pharmacology and in vivo study, it was found that SFJD exerted its therapeutic effects on BP by inhibiting the IL-17 pathway and JAK/STAT signaling pathway. This study provides new evidence for SFJD in treatment of BP.

Inula britannica ameliorates alcohol-induced liver injury by modulating SIRT1-AMPK/Nrf2/NF-jB signaling pathway

2024/11/13 14:19:56

Objective: Inula britannica is a traditional Chinese medicinal and functional food with various effects such as anti-liver injury, hypoglycemia, antioxidants, and anti-tumor. The aim of this study was to investigate the protective effects and mechanisms of the ethanolic extract of I. britannica (EEIB) on alcohol-induced liver injury in mice. Methods: Fifty-six female C57BL/6 mice were randomly divided into seven groups: control group (Con), ethanol feeding model group (EtOH), Silibinin positive treatment group (EtOH + Silibinin 100 mg/kg), EEIB treatment group (EtOH + EEIB 100, 200, and 400 mg/kg), and EEIB control group (EEIB 400 mg/kg). The National Institute on Alcohol Abuse and Alcoholism (NIAAA) ethanol-feeding model was used to study the effects of EEIB on alcohol-induced lipid metabolism, inflammation, oxidative stress, and fibril formation in mice by histopathological evaluation, immunofluorescence staining, Western blotting analysis and molecular docking. Results: EEIB reduced liver indices to different degrees to normal levels and improved liver morphology in mice. EEIB inhibited alcohol-induced liver injury by activating the sirtuin 1 (SIRT1)-adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in the liver of alcohol-fed mice, in which sesquiterpenes may be the potential active ingredients, and also down-regulated the expression of alpha-smooth muscle actin (a-SMA), collagen alpha (Collagen I), tumor necrosis factor-alpha (TNFa) and attenuated alcohol-induced liver injury. In addition, EEIB also activated the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, which alleviated alcohol-induced liver injury at the level of oxidative stress. Notably, the EEIB control group demonstrated that EEIB had no toxic effects in mice. EEIB reduced alcoholic liver injury in a dose-dependent manner. Its therapeutic efficacy was comparable to, if not better than, that of Silibinin when administered at a dose of 400 mg/kg. Conclusion: EEIB showed significant therapeutic effects on alcohol-induced liver injury in mice, and its mechanism of action was related to the SIRT1-AMPK, nuclear factor-kappa B (NF-jB), and Nrf2 signaling pathways, in which sesquiterpenes may be the potential active ingredients.

DNA metabarcoding uncovers fungal communities in Zingiberis Rhizoma

Chune Fan, Yanan Xu, Yufeng Li, Meihua Yang, Jianping Han, Xiaohui Pang * — 2024/11/13 14:19:56

Objective: Zingiberis Rhizoma (ZR, Ganjiang in Chinese), also known as dried ginger, is a popular spice and medicinal herb that has been used for several thousand years. However, ZR is easily contaminated by fungi and mycotoxin under suitable conditions, and might be hazardous to the health and safety of consumers, thus concerns about the herb’s safety have been raised. The aim of this study was to investigate the fungal community and the effects of collection areas and processing methods on the fungal community in ZR. Methods: A total of 18 ZR samples were collected from four provinces of China, and the samples were divided into four groups based on collecting sites. Meanwhile, the samples collected in Sichuan Province, China were divided into three groups based on the processing methods. We employed the Illumina MiSeq PE300 platform and targeted the internal transcribed spacer 2 (ITS2) sequences to investigate fungal contamination in ZR samples, and the difference in fungal community among the groups of different collection sites and processing methods. Results: All 18 samples were contaminated with fungi. Ascomycota was the dominant phyla, accounting for 34.46%–100% of the fungal reads. At the genus level, Candida, Diutina, and Aspergillus were the most dominant genera, with relative abundances of 0–98.37%, 0–99.82%, and 0–79.08%, respectively. Meanwhile, four potential toxigenic fungi and seven human pathogens were found. Furthermore, differences in the community composition of ZR samples from four collecting sites and three processing methods were observed. Conclusion: DNA metabarcoding provides a novel insight into fungal community diversity in ZR samples, providing references to ensure the sustainable utilization and quality research of ZR.

Herb induced liver injury by Xianling Gubao Tablets: A case assessed for causality using updated RUCAM and integrated evidence chain

Guohui Li a,*, Xiaohe Xiao b,* — 2024/4/23 14:27:20

Objective: A typical case of Xianling Gubao (XLGB) Tablets-induced liver injury was systematically studied in the clinic and the laboratory. Methods: A patient with herb-induced liver injury (HILI) and a history of taking XLGB Tablets before disease onset was engaged as the study subject, and the case was diagnosed according to the updated Roussel Uclaf Causality Assessment Method (RUCAM) and the integrated evidence chain (iEC) method recommended by the Guidelines for Diagnosis and Treatment of Herb-induced Liver Injury (HILI Guidelines). Results: Clinical history, biochemical indexes and imaging tests were used to exclude the influence of fundamental diseases and confusing liver diseases such as viral, alcoholic and autoimmune liver diseases on the diagnosis. Based on an investigation of the patient’s medication history, she was suspected to have HILI caused by XLGB Tablets, as the patient was only taking an oral preparation of XLGB Tablets, and the influence of other drugs on the diagnosis was excluded. This patient with alanine aminotransferase (ALT)≥ 3× upper limit of normal (ULN) and a calculated R of 6 was diagnosed with possible acute drug-induced hepatocellular injury. The relationship was considered ‘‘highly probable” (score of 9) using the updated RUCAM of 2016. Moreover, the fingerprint similarity between the preparation taken by the patient and a commercially available preparation was 0.99, suggesting that the patient was consuming XLGB Tablets rather than another drug. LC-MS technology and the Agilent Fake TCM-Drugs database were used to investigate the drug, and no chemical additions were found. Examination of the drug for pesticide residues, heavy metals, aflatoxins and other exogenous substances indicated compliance with the content limits of the Chinese Pharmacopoeia. Conclusion: In summary, the final diagnosis of XLGB-induced liver injury reached the clinical diagnosis of HILI and was acute severe hepatocellular injury type by the updated RUCAM and iEC. Therefore, this study provides scientific evidence regarding the causality evaluation of compound preparations of traditional Chinese medicines-induced liver injury.

Genetic diversity of food-medicinal Lycium spp. in China: Insights from chloroplast genome

2024/7/12 14:47:36

Objective: Goji (fruits of Lycium spp.) is commonly consumed as food and medicine. The increasing market demand for goji has led to its wide cultivation and broad breeding, which might cause loss of genetic diversity. This study aims to uncover the genetic diversity of the cultivated and wild Lycium. Methods: The chloroplast genome (CPG) of 34 accessions of Chinese food-medicinal Lycium spp., including the popular cultivars and their wild relatives, was re-sequenced and assembled, based on which the genetic diversity was evaluated. Results: Sequence structural comparison shows that CPG is comparatively conserved within species. Phylogenetic analysis indicates that CPG is sufficient for the discrimination of Lycium species; combined with nuclear ribosomal internal transcribed spacer (Nr ITS) sequences, materials with mixed genetic backgrounds can be identified. Nucleotide diversity analysis reveals that the modern cultivars are probably with a common maternal parent, while the wild accessions are with higher level of genetic diversity. Conclusion: For the first time this study reveals the intraspecies genetic diversity of Lycium spp. using CPG, highlighting the urgent conservation demand of wild genetic resources of Lycium. Our study also demonstrates that CPG provides crucial evidence for identification of Lycium species with mixed genetic backgrounds and highlights the importance of the wild relatives in genetic diversity conservation. This CPG-based technology will contribute to the sustainable development of medicinal plants broadly.

Analysis on patents of health care products with substances of medicine food homology in China

2024/7/12 14:47:36

Objective: The concept of substances of medicine food homology (SMFH) has garnered significant attention in recent years. This study conducts a systematic analysis of patent literature related to SMFH, and elucidates the development trends, technical hotspots, and the overall patent protection landscape of SMFH in China over the past two decades. Methods: The patent search focused on the SMFH varieties as the objects of inquiry, with retrieval conducted in patent databases. Subsequently, the acquired data underwent processing, analysis, and visualization. Results: While the technical threshold for pharmaceutical applications surpasses that of the food service sector, the former may assume a prominent role in the future. Research and development (R&D) activities in the southeast of China demonstrate robust activity than other regions. Colleges and scientific research institutions exhibit substantial advantages in patent applications compared with individuals and hold greater potential for future development. Conclusion: The findings of this patent analysis indicate that China’s SMFH industry are presently undergoing a transition from an extensive model to a high-quality model. The quality and technical standards of SMFH products are consistently improving. Consequently, there is a need for more stringent patent application requirements to align with the evolving development needs.

Amplifying protection against acute lung injury: Targeting both inflammasome and cGAS-STING pathway by Lonicerae Japonicae Flos-Forsythiae Fructus drug pair

2024/7/12 14:47:36

Objective: Acute lung injury (ALI) is characterized by inflammation and currently lacks an efficacious pharmacological intervention. The medicine combination of Lonicerae Japonicae Flos (LJF) and Forsythiae Fructus (FF) demonstrates combined properties in its anti infective, anti-inflammatory, and therapeutic effects, particularly in alleviating respiratory symptoms. In previous studies, Chinese medicine has shown promising efficacy in lipopolysaccharides (LPS)-induced ALI. However, there have been no reports of LJF and FF pairing for lung injury. The aim of this study is to compare the efficacy of herb pair Lonicerae Japonicae Flos-Forsythiae Fructus (LF) with LJF or FF alone in the treatment of ALI, and to explore whether LJF and FF have a combined effect in the treatment of lung injury, along with the underlying mechanism involved. Methods: A total of 36 mice were divided into six groups (control, model, LJF, FF, LF, dexamethasone) based on the treatments they received after undergoing sham-operation/LPS tracheal instillation. H&E staining and pulmonary edema indexes were used to evaluate lung injury severity. Alveolar exudate cells (AECs) were counted based on cell count in bronchoalveolar lavage fluid (BALF), and neutrophil percentage in BALF was measured using flow cytometry. Myeloperoxidase (MPO) activity in BALF was measured using enzyme-linked immunosorbent assay (ELISA), while the production of IL-1b, TNF-a, and IL-6 in the lung and secretion level of them in BALF were detected by quantitative polymerase chain reaction (qPCR) and ELISA. The effect of LJF, FF, and LF on the expression of caspase-1 and IL-1b proteins in bone marrow derived macrophages (BMDMs) supernatant was assessed using Western blot method under various inflammasome activation conditions. In addition, the concentration of IL-1b and changes in lactatedehydrogenase (LDH) release levels in BMDMs supernatant after LJF, FF, and LF administration, respectively, were measured using ELISA. Furthermore, the effects of LJF, FF and LF on STING and IRF3 phosphorylation in BMDMs were detected by Western blot, and the mRNA changes of IFN-b, TNF-a, IL-6 and CXCL10 in BMDMs were detected by qPCR. Results: LF significantly attenuated the damage to alveolar structures, pulmonary hemorrhage, and infiltration of inflammatory cells induced by LPS. This was evidenced by a decrease in lung index score and wet/dry weight ratio. Treatment with LF significantly reduced the total number of neutrophil infiltration by 75 % as well as MPO activity by 88 %. The efficacy of LF in reducing inflammatory factors IL-1b, TNF-a, and IL-6 in the lungs surpasses that of LJF or FF, approaching the effectiveness of dexamethasone. In BMDMs, the co-administration of 0.2 mg/mL of LJF and FF demonstrated superior inhibitory effects on the expression of nigericin-stimulated caspase-1 and IL-1b, as well as the release levels of LDH, compared to individual treatments. Similarly, the combination of 0.5 mg/mL LJF and FF could better inhibit the phosphorylation levels of STING and IRF3 and the production of IFN-b, TNF-a, IL-6, and CXCL10 in response to ISD stimulation. Conclusion: The combination of LJF and FF increases the therapeutic effect on LPS-induced ALI, which may be mechanistically related to the combined effect inhibition of cyclic-GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) and NOD-like receptor family protein 3 (NLRP3) inflammasomes pathways by LJF and FF. Our study provides new medicine candidates for the clinical treatment of ALI.

Integrative analysis of metabolome and transcriptome provides new insights into functional components of Lilii Bulbus

2024/7/12 14:47:37

Objective: Lilium brownii var. viridulum (LB) and L. lancifolium (LL) are the main sources of medicinal lily (Lilii Bulbus, Baihe in Chinese) in China. However, the functional components of these two species responsible for the treatment efficacy are yet not clear. In order to explore the therapeutic material basis of Lilii Bulbus, we selected L. davidii var. willmottiae (LD) only used for food as the control group to analyze the differences between LD and the other two (LB and LL). Methods: Metabolome and transcriptome were carried out to investigate the differences of active components in LD vs LB and LD vs LL. Data of metabolome and transcriptome was analysed using various analysis methods, such as principal component analysis (PCA), hierarchical cluster analysis (HCA), and so on. Differentially expressed genes (DEGs) were enriched through KEGG and GO enrichment analysis. Results: The PCA and HCA of the metabolome indicated the metabolites were clearly separated and varied greatly in LL and LB contrasted with LD. There were 318 significantly differential metabolites (SDMs) in LD vs LB group and 298 SDMs in LD vs LL group. Compared with LD group, the significant upregulation of steroidal saponins and steroidal alkaloids were detected both in LB and LL groups, especially in LB group. The HCA of transcriptome indicated that there was significant difference in LB vs LD group, while the difference between LL and LD varied slightly. Additionally, 47 540 DEGs in LD vs LB group and 18 958 DEGs in LD vs LL group were identified. Notably, CYP450s involving in the biosynthesis of steroidal saponins and steroidal alkaloids were detected, and comparing with LD, CYP724, CYP710A, and CYP734A1 in LB and CYP90B in LL were all up-regulated. Conclusion: This study suggested that steroidal saponins and steroidal alkaloids maybe the representative functional components of Lilii Bulbus, which can provide new insights for Lilii Bulbus used in the research and development of classic famous formula.

HPLC combined with chemometrics for quality control of Huamoyan Granules or Capsules

2024/7/12 14:47:37

Objective: Huamaoyan Granules (HMYG) and Huamaoyan Capsules (HMYC) are Chinese patent medicines with different dosage forms of the same prescription. Due to the different preparation process, the chemical composition of these Chinese patent medicines varies greatly among different forms, but there were few studies on the difference comparison and quality control of them. In order to improve the effectiveness and safety in its clinical application, an idea combining high performance liquid chromatography (HPLC) and chemometrics was put forward to study the quality control of Chinese patent medicines in different dosage forms of the same prescription. Methods: The differential markers of HMYG and HMYC were explored based on HPLC fingerprint and chemometrics including orthogonal projections to latent structures-discriminant analysis (OPLS-DA), principal component analysis (PCA), and hierarchical cluster analysis (HCA). Finally, the quantitative analysis method of related components was established by HPLC. Results: A quality control method for HMYG and HMYC was established. Firstly, the chemical components of HMYG and HMYC were systematically analyzed by HPLC fingerprinting. Further exploration showed that there were 20 characteristic peaks and 57 common peaks. Then, the potential differential markers between HMYG and HMYC were explored by chemometrics, and the differential markers were screened after intersection with the 20 characteristic peaks. Finally, HPLC quantitative analysis methods for nine components were established, including seven differential markers (neochlorogenic acid, protocatechualdehyde, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rosmarinic acid and salvianolic acid A). The results of HPLC quantitative analysis showed that the contents of eight components in HMYG and HMYC samples were significantly different. According to the above results, the differential markers between HMYG and HMYC screened based on HPLC fingerprint and chemometrics can effectively characterize the differences between the two dosage forms. Conclusion: The present work provides a rapid and effective method for routine quality evaluation and control of HMYG and HMYC. This work also provides feasible methods for the quality evaluation and control of Chinese patent medicines with different dosage forms of the same prescription.

Comparative pharmacokinetics of six components in normal and rheumatoid arthritis rats after intragastrical administration of Qianghuo Shengshi Decoction granules by LC-MS/MS

2024/7/12 14:47:37

Objective: To investigate the plasma pharmacokinetics of six representative components (nodakenin, osthole, 5-O-methylvisammioside, ferulic acid, liquiritigenin, and liquiritin), which were the ingredients of Qianghuo Shengshi Decoction (QSD) granules, in normal and rheumatoid arthritis (RA) rats administrated QSD granules intragastrically. Methods: A rapid and accurate ultra-high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of six components in plasma, and it showed a good specificity, linearity, intra-day and inter-day precision, intra-day and inter-day accuracy, extraction recovery, stability, and the less matrix effect. Results: The validated LC-MS/MS method was successfully used to compare the plasma pharmacokinetics of six ingredients between normal and RA rats after intragastrical administration of QSD granules and differences in the pharmacokinetics were found in two types of rats. The absorption rate in the RA rats was lower for nodakenin, osthole, 5-O-methylvisammioside, liquiritigenin and liquiritin than in the normal group, while the absorption rate of ferulic acid remained constant in two groups. In comparison with the normal rats, the exposure concentration of nodakenin was higher and that of other five components except for nodakenin was lower under pathological conditions. Additionally, the absorptive amount of nodakenin, osthole, 5-O-methylvisammioside and liquiritin was increased and that of ferulic acid and liquiritigenin was reduced in the RA rats than in the normal rats. Compared with the normal rats, the retention time of nodakenin, ferulic acid and liquiritin was reduced in vivo, whereas the retention time of osthole, 5-O methylvisammioside and liquiritigenin was raised in the body for the RA rats. In contrast to the normal rats, the data demonstrated an increase in the elimination velocity of nodakenin and a decrease in the elimination velocity of the other five components except for nodakenin in the pathological state. Conclusion: This study showed that the pharmacokinetic behavior of the six components, nodakenin, osthole, 5-O-methylvisammioside, ferulic acid, liquiritigenin, and liquiritin, is different in vivo between normal and pathological states of rats, and this research provided the necessary experimental data to explain the pharmacokinetics of QSD granules in both normal and pathological states and provide some references for its clinical application at some level.

Identification and characterization of chemical constituents in Mahuang Guizhi Decoction and their metabolites in rat plasma and brain by UPLC-Q-TOF/MS

2024/7/12 14:47:37

Objective: Mahuang Guizhi Decoction (MGD), an essential herbal pair in traditional Chinese medicine, is able to release cold, fever and asthma, mainly containing alkaloids, flavonoids, phenylpropanoids and amino acids. However, the absorption and distribution of these four category compounds in vivo still remained unclearly. Methods: In our research, we utilized UPLC-Q-TOF-MS technique to identify the constituents within MGD, as well as the prototypes of MGD and their metabolites absorbed in plasma and brain. We further profiled the drug-time curve of prototypes and metabolites of MGD both in plasma and brain. Results: Our results showed that 105 constituents were characterized in MGD. Thirty of them could be absorbed into blood, and ten of them could be distributed into brain. We also discovered eight new bio-transformed metabolites in blood, and a half of which could pass through the blood–brain barrier. In addition, all components detected in vivo could be absorbed and distributed immediately. Conclusion: These findings provide an approachable method to analyze the potential bio-active compounds in MGD and their in vivo behaviors, which could promote the efficacious material basis study of MGD and the security of clinical utilization.

Constituents from leaves of Macaranga hemsleyana

2024/7/12 14:47:37

Objective: To study constituents of the leaves of Macaranga hemsleyana, and evaluate their inhibitory effects against NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome activation, and antiproliferative activity. Methods: The constituents were isolated and purified by column chromatography on MCI gel CHP20P/P120, silica gel, Sephadex LH-20, and HPLC. The structures of compounds were determined by 1D, 2D NMR, and HR-ESI-MS data. The inhibitory effect of compounds on inflammasome activation was determined by lactate dehydrogenase (LDH) procedure. The antiproliferative activity was evaluated using MTT assay. Results: The study led to the isolation of 23 compounds, including one new compound, identified as (2Z)-3-[4-(b-D-glucopyranosyloxy)-20-hydroxy-50-methoxyphenyl]-2-propenoic acid (1), together with 22 known compounds recognized as 1,4-dihydro-4-oxo-3 pyridinecarbonitrile (2), methyl 4-methoxynicotinate (3), 4-methoxynicotinonitrile (4), 1-(3-O-b-D-glucopyranosyl-4,5-dihydroxyphenyl)-ethanone (5), neoisoastilbin (6), isoastilbin (7), aromadendrin (8), neoastilbin (9), astilbin (10), quercitrin (11), neoschaftoside= (12), apigenin 6,8-bis-C-a-L-arabinoside (13), vitexin (14), bergenin (15), scopoletin (16), glucopyranoside salicyl (17), koaburside (18), benzyl b-D-glucoside (19), icariside B5 (20), roseoside (21), loliolide (22), and adenosine (23). The tested compounds did not show LDH inhibition nor antiproliferative activity. Conclusion: Compound 1 was a new glycoside. Compounds 2 and 3 were obtained for the first time from natural source. The 22 known compounds constituted of alkaloids (2–4, 23), phenolics (5, 15, 17, 18), flavonoids (6–14), coumarin (16), benzyl glycoside (19), and norsesquiterpenes (20–22). All the compounds, 1–23, were revealed from M. hemsleyana for the first time. This is the initial uncovering of molecules 1–10, 12, 13, 17–19, and 23 from the genus Macaranga. The isolated compounds, 11, 14–16, and 20–22 established taxonomic classification of M. hemsleyana in Euphorbiaceae family. Flavonoids were outstanding as chemosystematic markers of Macaranga genus.

Arbuscular mycorrhiza and plant growth promoting endophytes facilitates accumulation of saponin under moderate drought stress

2024/4/23 14:27:20

Objective: Paris polyphylla var. yunnanensis, one of the important medicinal plant resources in Yunnan, China, usually takes 6–8 years to be harvested. Therefore, it is urgent to find a method that can not only shorten its growth years, but also improve its quality. In this study, we examined the effects of a combination treatment of arbuscular mycorrhizal fungi (AMF) and plant growth-promoting endophytes (PGPE) and drought stress on the accumulation of saponins in it. Methods: P. polyphylla var. yunnanensis was infected with a mixture of AMF and PGPE under drought stress. The content of saponins, as well as morphological, physiological, and biochemical indicators, were all measured. The UGTs gene related to saponin synthesis was obtained from transcriptome data by homologous comparison, which were used for RT-PCR and phylogenetic analysis. Results: Regardless of water, AMF treatment could infect the roots of P. polyphylla var. yunnanensis, however double inoculation with AMF and PGPE (AMF + PGPE) would reduce the infection rate of AMF. Plant height, aboveground and underground fresh weight did not differ significantly between the single inoculation AMF and the double inoculation treatment under different water conditions, but the inoculation treatment significantly increased the plant height of P. polyphylla var. yunnanensis compared to the noninoculation treatment. Single inoculation with AMF considerably increased the net photosynthetic rate, stomatal conductance, and transpiration rate of P. polyphylla var. yunnanensis leaves under various water conditions, but double inoculation with AMF + PGPE greatly increased the intercellular CO2 concentration and chlorophyll fluorescence parameter (Fv/Fm). Under diverse water treatments, single inoculation AMF had the highest proline content, whereas double inoculation AMF + PGPE may greatly improve the amount of abscisic acid (ABA) and indoleacetic acid (IAA) compared to normal water under moderate drought. Double inoculation AMF + PGPE treatment improved the proportion of N, P, and K in the rhizome of P. polyphylla var. yunnanensis under various water conditions. Under moderate drought stress, AMF + PGPE significantly enhanced the contents of P. polyphylla var. yunnanensis saponins I, II, VII, and total saponins as compared to normal water circumstances. Farnesyl diphosphate synthase (FPPS), Geranyl pyrophosphate synthase (GPPS), Cycloartenol synthase (CAS), and Squalene epoxidase (SE1) were the genes that were significantly up-regulated at the same time. The amount of saponins was favorably linked with the expression of CAS, GPPS, and SE1. Saponin VI content and glycosyl transferase (UGT) 010922 gene expression were found to be substantially associated, as was saponin II content and UGT010935 gene expression. Conclusion: Under moderate drought, AMF + PGPE was more conducive to the increase of hormone content, nutrient absorption, and total saponin content in P. polyphylla var. yunnanensis, and AMF + PGPE could up regulate the expression of key genes and UGTs genes in one or more steroidal saponin synthesis pathways to varying degrees, thereby stimulating the synthesis and accumulation of steroidal saponins in the rhizome of P. polyphylla var. yunnanensis. The combination of AMF and PGPE inoculation, as well as adequate soil drought, reduced the buildup of saponins in P. polyphylla var. yunnanensis and increased its quality.

Metabolites isolated from Penicillium HDS-Z-1E, an endophytic fungal strain isolated from Taxus cuspidata and their activation effect of catalase

2024/4/23 14:27:20

Objective: To study the compounds isolated from Penicillium HDS-Z-1E, an endophytic fungal strain isolated from Taxus cuspidata and their activation effect of catalase (CAT). Methods: The chemical constituents were isolated from Penicillium HDS-Z-1E, by using silica gel, Sephadex LH-20 and HPLC. The structural elucidations of five metabolites were elucidated on the basis of spectroscopic including 1H NMR, 13C NMR, HMBC and HSQC. Their activation sites of catalase have been investigated by molecular docking. Results: Five metabolites, compounds (1–5) were isolated from Penicillium HDS-Z-1E and identified as 4-hydroxy-4-methyltetrahydro-2H-pyran-2-one (1), 4-hydroxymethyl-5, 6-dihydro-pyran-2-one (2), 5, 6-dihydro-2-oxo-2H-pyran-4-carboxylic (3), N-acetyl hydrazinobenzoic acid (4), and methyl 2-(2, 5-dihydroxyphenyl) acetate (5). Conclusion: Compound 3 is a new compound. Compounds 3 and 4 may have potential activators of catalase, providing a theoretical basis for the development of CAT activators.

A new tetracyclic triterpenoid from endophytic fungus Fusarium sporotrichioides

Yajing Wang, Chunyue Liu, Yihui Yang, Chang Li *, Yuehu Pei * — 2024/4/23 14:27:20

Objective: To isolate bioactive compounds from the endophytic fungus Fusarium sporotrichioides isolated from Rauwolfia yunnanensis, and investigate their pharmacological activities. Methods: The chemical constituents were isolated and purified by combining with ODS column chromatography, silica gel column chromatography and by performing semipreparative HPLC. Their structures were established on the basis of 1D NMR (1H NMR and 13C NMR) and 2D NMR (1H–1H COSY, HSQC, HMBC and NOESY), as well as HRESIMS and comparison with literature data. In addition, the absolute configuration of compound 1 was determined by calculated ECD data. Results: One previously undescribed tetracyclic triterpenoid derivative, named as integracide L (1), 12α-acetoxy-4,4-dimethyl-24-methylene-5α-cholesta-8,14-diene-2α,3β,11β-triol (2), 12α-acetoxy-4,4-dimethyl-24-methylene-5α-cholesta-8-momoene-2α,3β,11β-triol (3), 12α-acetoxy-4,4-dimethyl-24-methylene-5α-cholesta-8,14-diene-3β,11β-triol (4), and 12α-acetoxy-4,4-dimethyl-24-methylene-5α-cholesta-8-momoene-3β,11β-triol (5) were isolated from F. sporotrichioide. Moreover, compound 1 was rare tetracyclic triterpenoid with single methyl replacement at C-4 position. Conclusion: Compound 1 was a new tetracyclic triterpenoid isolated from the endophytic fungus F. sporotrichioides. In addition, compound 2 could inhibit the growth of three different human cancer cells significantly. Compounds 3 and 5 were found to possess better cytotoxic activities on HepG-2 cells than the other compounds, with IC50 values of (2.8 ± 0.1) and (6.3 0.3)μmol/L respectively.

New indole derivatives from endophytic fungus Colletotruchum sp. HK-08 originated from leaves of Nerium indicum

2024/4/23 14:27:20

Objective: To study secondary metabolites from endophytic fungus Colletotruchum sp. HK-08 originated from the leaves of Nerium indicum. Methods: The compounds were isolated by various column chromatographic techniques, and their structures were elucidated by spectroscopic techniques [high resolution electrospray ionization mass spectroscopy (HRESIMS), one-dimensional (1D) and two dimensional (2D) nuclear magnetic resonance spectroscopy (NMR)], as well as comparison with literature data. The Ellman method was used to determine the acetylcholinesterase (AChE) inhibitory activity. Results: Four indole derivatives were identified from Colletotruchum sp. HK-08, including 60-hydroxymonaspiloindole (1), 2-(2-oxoindolin-3-yl)ethyl 2-(4-hydroxyphenyl) acetate (2), 2-(2-oxoindolin-3-yl)ethyl 2-(2-hydroxyphenyl)acetate (3), and monaspiloindole (4). Compound 4 presented weak AChE inhibitory activity with IC50 value of (69.30 ± 6.27) μmol/L [tacrine as the positive control, with IC50 value of (0.61 ± 0.07) μmol/L]. Conclusion: Compounds 1–3 were new compounds, and compound 4 had weak AChE inhibitory activity.

Protective role of fruits of Rosa odorata var. gigantea against WIRS induced gastric mucosal injury in rats by modulating pathway related to inflammation, oxidative stress and apoptosis

2024/4/23 14:27:20

Objective: Rosa odorata var. gigantea is a popular medicinal plant. Some studies have demonstrated that ethanolic extract of the fruits of R. odorata var. gigantea (FOE) has gastroprotective properties. The aim of this study was to investigate the gastroprotective activity of FOE on water immersion restrained stress (WIRS)-induced gastric mucosal injury in a rat model and elucidate the possible molecular mechanisms involved. Methods: A rat stress ulcer model was established in this study using WIRS. After rats were treated with FOE orally for 7 d, the effect of FOE treatment was analyzed by hematoxylin and eosin (H&E) staining, and the changes of inflammatory factors, oxidative stress factors, and gastric-specific regulatory factors and pepsin in the blood and gastric tissues of rats were examined by ELISA assay. Molecular mechanism of FOE was investigated by immunohistochemical assay and Western blot. Results: Compared with the WIRS group, FOE could diminish both the macroscopic and microscopic pathological morphology of gastric mucosa. FOE significantly preserved the antioxidants glutathione peroxidase (GSH-PX), superoxide dismutase (SOD) and catalase (CAT) contents; anti-inflammatory cytokines interleukin-10 (IL-10) and prostaglandin E2 (PGE2) levels as well as regulatory factors tumor necrosis factor-α (TGF-α) and somatostatin (SS) contents, while decreasing malondialdehyde (MDA), nitric oxide synthase (iNOS), tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), gastrin (GAS) and endothelin (ET) levels. Moreover, FOE distinctly upregulated the expression of Nrf2, HO-1, Bcl2 and proliferating cell nuclear antigen (PCNA). In addition, FOE activated the expression of p-EGFR and downregulated the expression of NF-jB, Bax, Cleaved-caspase-3, Cyto-C and Cleaved-PARP1, thus promoting gastric mucosal cell survival. Conclusion: The current work demonstrated that FOE exerted a gastroprotective activity against gastric mucosal injury induced by WIRS. The underlying mechanism might be associated with the improvement of anti-inflammatory, anti-oxidation and anti-apoptosis systems.

Huangqi Guizhi Wuwu Decoction suppresses inflammation and bone destruction in collagen-induced arthritis mice

2024/4/23 14:27:20

Objective: Rheumatoid arthritis (RA) is a chronic inflammatory and destructive arthritis, characterized by inflammatory infiltration and bone destruction. Huangqi Guizhi Wuwu Decoction (HGWD) is traditional Chinese medicine, which has been applied in the treatment of RA in clinical. The aim of this study was to investigate the therapeutic effect of HGWD on collagen-induced arthritis (CIA) mouse model. Methods: DBA/1J female mice were used to establish the collagen-induced arthritis (CIA) model. HGWD was administered intragastrically once a day for four weeks starting on the 22nd day after the first immunization. The body weight, hind paw thickness and clinical score were measured every five days. Gait analysis, histopathological staining, enzyme-linked immunosorbent assay (ELISA), ultrasound imaging and micro-computed tomography imaging were performed to determine the effects of HGWD treatment on inflammation and bone structure in this model. Moreover, Real-time PCR and Western blot analysis were used to detect inflammatory factors mRNA and protein levels after HGWD intervention in RAW 264.7 cells. Results: HGWD attenuated symptoms of arthritis, suppressed inflammatory synovium area and the serum levels of inflammatory factors, inhibited joint space enlargement in the knee and ankle joints, reduced numbers of osteoclasts, protected bone destruction, as well as improved motor function. HGWD decreased the expression of mRNA for inflammatory factors and the protein expression levels of p-NF-κB and IL-17. Conclusion: These results suggested that HGWD suppresses inflammation, attenuates bone erosion and maintains motor function in collagen-induced arthritis mice.

Pleiotropic attenuating effect of Ginkgo biloba against isoprenalineinduced myocardial infarction via improving Bcl-2/mTOR/ERK1/2/Na+, K+-ATPase activities

2024/4/23 14:27:20

Objective: Myocardial infarction (MI) is linked to an imbalance in the supply and demand of blood oxygen in the heart muscles. Beta-blockers and calcium antagonists are just two of the common medications used to treat MI. However, these have reportedly been shown to be either ineffective or to have undesirable side effects. Extract of Ginkgo biloba leaves (GBE), a Chinese herbal product offers special compatibility benefits in therapeutic settings relating to inflammatory diseases and oxidative stress. In order to better understand how GBE affects MI in rats insulted by isoprenaline (ISO), the current study was designed. Methods: The heart weight index, serum lipid profile, cardiac marker enzymes, endogenous antioxidants [catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), nitrites and malondialdehyde (MDA)], inflammatory mediators [tumour necrosis factor alpha (TNF-α) and interleukin-6 (IL-6)], immunohistochemical expressions of B-cell lymphoma factor-2 (Bcl-2), extracellular signal-regulated kinase (ERK1/2), and mammalian target of rapamycin (mTOR) and histopathological analysis were used to assess the cardioprotective properties of GBE. Results: The findings showed that GBE effectively attenuated myocardial infarction by boosting the body’s natural antioxidant defense system and reducing the release of inflammatory cytokines as well as heart injury marker enzymes. The expression of Bcl-2, ERK1/2 and mTOR was increased while the histomorphological alterations were reversed. Conclusion: The cardioprotective effects of GBE may be due to a mechanism involving increased Bcl-2/ mTOR/ERK1/2/Na+, K+-ATPase activity.

Chemical profiling of principle active and toxic constituents in herbs containing aristolochic acids

2024/4/23 14:27:20

Objective: To clear the amounts of the principal active/toxic components in herbs containing aristolochic acids (HCAAs), which are still used as medicine and/or seasoning in many ethnic minority areas of China. Methods: In this study, six major active and toxic components in HCAAs were extracted with ultrasonic extraction. With 6-O-methyl guanosine as internal standard, the target compounds were analyzed qualitatively and quantitatively by using ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) with multiple reaction monitoringinformation dependent acquisition-enhanced production ion scanning mode (MRM-IDA-EPI) combined with dynamic background subtraction (DBS) function. Results: The method showed good linearity in the linear range of the six analytes. The limit range of detection was from 0.01 ng/mL to 0.27 ng/mL. All of the detection repeatability, extraction repeatability and accuracy of the method were good. After extraction, the samples remained stable at 15℃ within 24 h. Six analytes were all found in samples except aristolactam (AL) in sample 2, and the contents varied greatly. The contents of these compounds decreased in fruits, leaves and stems of Aristolochia delavayi successively. Conclusion: This method has the advantages of less sample dosage, simple operation, short analysis cycle, high sensitivity, specificity and accuracy. It laid a good foundation for guiding the safety of HCAAs, the indepth study of pharmacological and toxicological effects and the scientific and standardized processing and compatibility of HCAAs.

Chuanxiong Rhizoma extracts prevent liver fibrosis via targeting CTCF-c-MYC-H19 pathway

2024/3/11 8:20:48

Objective: Hepatic fibrosis has been widely considered as a conjoint consequence of almost all chronic liver diseases. Chuanxiong Rhizoma (Chuanxiong in Chinese, CX) is a traditional Chinese herbal product to prevent cerebrovascular, gynecologic and hepatic diseases. Our previous study found that CX extracts significantly reduced collagen contraction force of hepatic stellate cells (HSCs). Here, this study aimed to compare the protection of different CX extracts on bile duct ligation (BDL)-induced liver fibrosis and investigate plausible underlying mechanisms. Methods: The active compounds of CX extracts were identified by high performance liquid chromatography (HPLC). Network pharmacology was used to determine potential targets of CX against hepatic fibrosis. Bile duct hyperplasia and liver fibrosis were evaluated by serologic testing and histopathological evaluation. The expression of targets of interest was determined by quantitative real-time PCR (qPCR) and Western blot. Results: Different CX extracts were identified by tetramethylpyrazine, ferulic acid and senkyunolide A. Based on the network pharmacological analysis, 42 overlap targets were obtained via merging the candidates targets of CX and liver fibrosis. Different aqueous, alkaloid and phthalide extracts of CX (CXAE, CXAL and CXPHL) significantly inhibited diffuse severe bile duct hyperplasia and thus suppressed hepatic fibrosis by decreasing CCCTC binding factor (CTCF)-c-MYC-long non-coding RNA H19 (H19) pathway in the BDL induced mouse model. Meanwhile, CX extracts, especially CXAL and CXPHL also suppressed CTCF-c-MYC-H19 pathway and inhibited ductular reaction in cholangiocytes stimulated with taurocholate acid (TCA), lithocholic acid (LCA) and transforming growth factor beta (TGF-b), as illustrated by decreased bile duct proliferation markers. Conclusion: Our data supported that different CX extracts, especially CXAL and CXPHL significantly alleviated hepatic fibrosis and bile duct hyperplasia via inhibiting CTCF-c-MYC-H19 pathway, providing novel insights into the anti-fibrotic mechanism of CX.

Magnolol and 5-fluorouracil synergy inhibition of metastasis of cervical cancer cells by targeting PI3K/AKT/mTOR and EMT pathways

Yuanyuan Chen a, Shanshan Chen a, Kaiting Chen a, Lanfang Ji a, Shuna Cui a,b,* — 2024/3/11 8:20:48

Objective: This study is designed to investigate the mode of action of the synergistic effect of 5-fluorouracil (5-FU) and magnolol against cervical cancer. Methods: Network pharmacological approach was applied to predict the molecular mechanism of 5-FU combined with magnolol against cervical cancer. CCK-8 assay, colony formation assay, immunofluorescence staining, adhesion assay, wound healing mobility assay, cell migration and invasion assay and Western blot analysis were conducted to validate the results of in silico study. Results: Phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway was identified as the key pathway in silico study. The experimental results showed that 5-FU combined with magnolol strongly inhibited cervical cancer cell proliferation, induced the morphological change of HeLa cells by down-regulating the expression of a-actinin, tensin-2 and vinculin. Moreover, magnolol enhanced inhibitory effect of 5-FU on the cell adhesion, migration and invasion. The phosphorylation of AKT and PI3K and the expression of mTOR were strongly inhibited by the combination of 5-FU and magnolol. Moreover, the expression of E-cadherin and b-catenin was upregulated and the expression of Snail, Slug and vimentin was down regulated by the 5-FU together with magnolol. Conclusion: Taken together, this study suggests that 5-FU combined with magnolol exerts a synergistic anti-cervical cancer effect by regulating the PI3K/AKT/mTOR and epithelial-mesenchymal transition (EMT) signaling pathways.

Anemoside B4 inhibits SARS-CoV-2 replication in vitro and in vivo

2024/3/11 8:20:48

Objective: Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARSCoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro. Methods: The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining. Results: The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-b response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism. Conclusion: Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.

Jatonik polyherbal mixture induced rat liver MMPT pore opening in normal Wistar rat: In vitro and in vivo studies

2024/3/11 8:20:48

Objective: To assess acute toxicity, the in vitro and in vivo effects of methanol and ethyl acetate extracts (JME and JEE) of Jatonik polyherbal mixture on some mitochondria-related parameters and their effect on the activity of some liver enzymes. Methods: Acute toxicity of JME and JEE was determined using Lorke’s method. In vitro and in vivo opening of the mitochondrial membrane permeability transition pore (MMPT pore) was spectrophotometrically assayed. Production of malondialdehyde (MDA) as an index of lipid peroxidation and the activity of mitochondrial ATPase was evaluated in vitro and in vivo and the effect of JME and JEE on the activity of liver enzymes such as alkaline phosphatase (ALP), aspartate and alanine aminotransferase (AST and ALT) and gammaglutamyl transferase (GGT)was also investigated. Results: JME had an LD50 of 3808 mg/kg b.w whereas JEE had an LD50 greater than 5000 mg/kg b.w. of rats. After the rats have been fed with both extracts, a photomicrograph of a piece of liver tissue showed no apparent symptoms of toxicity. From the in vitro and in vivo studies, both extracts prompted intact mitochondria to open their MMPT pores. When compared to the control, lipid peroxide product release and ATPase activity were significantly increased (P < 0.05) in vitro and in vivo. The activities of AST, ALT, and GGT were all reduced at 50 mg/kg when treated with JME, but the activity of AST was considerably enhanced when treated with JEE (P < 0.05). The results revealed that both JME and JEE of the Jatonik polyherbal mixture had low toxicity, profound MMPTpore induction, and enhanced ATPase activity, but an increased MDA production. Conclusion: Jatonik extracts may be a promising target for drug development in diseases where there is dysregulation of apoptosis, however, further studies are needed to better clarify the molecular mechanism involved in these phenomena.

Effect of Rhei Radix et Rhizoma and Eupolyphaga Steleophaga on liver protection mechanism based on pharmacokinetics and metabonomics

2024/3/11 8:20:48

Objective: Based on metabonomics technology of high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) and hydrogen nuclear magnetic resonance spectroscopy (1H NMR), the pharmacokinetic characteristics and therapeutic mechanism of Rhei Radix et Rhizoma (RhRR, Dahuang in Chinese), Eupolyphaga Steleophaga (EuS, Tubiechong in Chinese) combined with RhRR acting on acute liver injury were explored. Methods: Models of acute liver injury were established, and the pharmacokinetic methods of five components of RhRR-EuS in rats were found by HPLC-MS/MS. The liver tissues of different groups of mice were analyzed by 1H NMR spectroscopy combined with multivariate statistical analysis to investigate the metabolomics of RhRR-EuS and RhRR. Results: Pharmacokinetic results showed there were different levels of bimodal phenomenon in different groups, and the absorption of free anthraquinone in RhRR increased after compatibility with EuS. In addition, the pathological state of acute liver injury in rats can selectively promote the absorption of emodin, chrysophanol, physcion and aloe emodin. Through 15 differential metabolites in the liver tissue of acute liver injury mice, it was revealed that RhRR-EuS and RhRR could protect the liver injury by regulating the metabolism of glutamine and glutamic acid, alanine, aspartic acid and glutamic acid, and phosphoinositide. However, the regulation of RhRR was weaker than that of RhRR-EuS. Conclusion: For the first time, we studied the pharmacokinetics and metabolomics differences of RhRREuS and RhRR in rats and mice with acute liver injury, in order to provide theoretical reference for clinical treatment of liver disease by DHZCP.

Effects of Xiaoyao San on exercise capacity and liver mitochondrial metabolomics in rat depression model

2024/3/11 8:20:48

Objective: This study aimed to investigate the therapeutic effects of Xiaoyao San (XYS), a herbal medicine formula, on exercise capacity and liver mitochondrial metabolomics in a rat model of depression induced by chronic unpredictable mild stress (CUMS). Methods: A total of 24 male SD rats were randomly divided into four groups: control group (C), CUMS control group (M), Venlafaxine positive treatment group (V), and XYS treatment group (X). Depressive behaviour and exercise capacity of rats were assessed by body weight, sugar-water preference test, open field test, pole test, and rotarod test. The liver mitochondria metabolomics were analyzed by using liquid chromatography-mass spectrometry (LC-MS) method. TCMSP database and GeneCards database were used to screen XYS for potential targets for depression, and GO and KEGG enrichment analyses were performed. Results: Compared with C group, rats in M group showed significantly lower body weight, sugar water preference rate, number of crossing and rearing in the open field test, climbing down time in the pole test, and retention time on the rotarod test (P < 0.01). The above behaviors and exercise capacity indices were significantly modulated in rats in V and X groups compared with M group (P < 0.05, 0.01). Compared with C group, a total of 18 different metabolites were changed in the liver mitochondria of rats in M group. Nine different metabolites and six metabolic pathways were regulated in the liver mitochondria of rats in X group compared with M group. The results of network pharmacology showed that 88 intersecting targets for depression and XYS were obtained, among which 15 key targets such as IL-1b, IL-6, and TNF were predicted to be the main differential targets for the treatment of depression. Additionally, a total of 1 553 GO signaling pathways and 181 KEGG signaling pathways were identified, and the main biological pathways were AGE-RAGE signaling pathway, HIF-1 signaling pathway, and calcium signaling pathway. Conclusion: XYS treatment could improve depressive symptoms, enhance exercise capacity, positively regulate the changes of mitochondrial metabolites and improve energy metabolism in the liver of depressed rats. These findings suggest that XYS exerts antidepressant effects through multi-target and multi-pathway.

DNA metabarcoding analysis of fungal community on surface of four root herbs

Yujie Dao, Jingsheng Yu, Meihua Yang, Jianping Han, Chune Fan, Xiaohui Pang * — 2024/3/11 8:20:48

Objective: Angelicae Sinensis Radix (ASR, Danggui in Chinese), Cistanches Herba (CH, Roucongrong in Chinese), Ginseng Radix et Rhizoma (PG, Renshen in Chinese), and Panacis Quinquefolii Radix (PQ, Xiyangshen in Chinese), widely used as medicine and dietary supplement around the world, are susceptible to fungal and mycotoxin contamination. In this study, we aim to analyze their fungal community by DNA metabarcoding. Methods: A total of 12 root samples were collected from three main production areas in China. The samples were divided into four groups based on herb species, including ASR, CH, PG, and PQ groups. The fungal community on the surface of four root groups was investigated through DNA metabarcoding via targeting the internal transcribed spacer 2 region (ITS2). Results: All the 12 samples were detected with fungal contamination. Rhizopus (13.04%-74.03%), Aspergillus (1.76%-23.92%), and Fusarium (0.26%-15.27%) were the predominant genera. Ten important fungi were identified at the species level, including two potential toxigenic fungi (Penicillium citrinum and P. oxalicum) and eight human pathogenic fungi (Alternaria infectoria, Candida sake, Hyphopichia burtonii, Malassezia globosa, M. restricta, Rhizopus arrhizus, Rhodotorula mucilaginosa, and Ochroconis tshawytschae). Fungal community in ASR and CH groups was significantly different from other groups, while fungal community in PG and PQ groups was relatively similar. Conclusion: DNA metabarcoding revealed the fungal community in four important root herbs. This study provided an important reference for preventing root herbs against fungal and mycotoxin contamination.

Hypoglycemic activities of flowers of Xanthoceras sorbifolia and identification of anti-oxidant components by off-line UPLC-QTOF-MS/MS-free radical scavenging detection

2024/3/11 8:20:48

Objective: To identify phytochemical constituents present in the extract of flowers of Xanthoceras sorbifolia and evaluate their anti-oxidant and anti-hyperglycemic capacities. Methods: The AlCl3 colorimetric method and Prussian Blue assay were used to determine the contents of total flavonoids and total phenolic acids in extraction layers, and the bioactive layers was screened through anti-oxidative activity in vitro. The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC BEH C18 column (2.0 mm × 150 mm, 5 μm) were used to identify the ingredients. And antioxidative ingredients were screened by off-line UPLC-QTOF-MS/MS-free radical scavenging. The ameliorative role of it was further evaluated in a high-fat, streptozotocin-induced type 2 diabetic rat model and the study was carried out on NADPH oxidase (PDB ID: 2CDU) by molecular docking. Results: Combined with the results of activity screening in vitro, the anti-oxidative part was identified as the ethyl acetate layer. A total of 24 chemical constituents were identified by liquid chromatographymass spectrometry in the ethyl acetate layer and 13 main anti-oxidative active constituents were preliminarily screened out through off-line UPLC-QTOF-MS/MS-free radical scavenging. In vivo experiments showed that flowers of X. sorbifolia could significantly reduce the blood glucose level of diabetic mice and alleviate liver cell damage. Based on the results of docking analysis related to the identified phytocompounds and oxidase which involved in type 2 diabetes, quercetin 3-O-rutinoside, kaempferol-3-Orhamnoside, isorhamnetin-3-O-glucoside, and isoquercitrin showed a better inhibitory profile. Conclusion: The ethyl acetate layer was rich in flavonoids and phenolic acids and had significant antioxidant activity, which could prevent hyperglycemia. This observed activity profile suggested X. sorbifolia flowers as a promising new source of tea to develop alternative natural anti-diabetic products with a high safety margin.

Chemical components in cultivated Cordyceps sinensis and their effects on fibrosis

Zhonghua Dong a, Xiao Sun b,c,d,e,* — 2024/3/11 8:20:48

Objective: Cultivated Cordyceps sinensis powder has been used as clinical drug and healthy food to nourish the lung and kidney, which solves the problem of serious shortage of wild C. sinensis. This study aims to explore the chemical components and compared their anti-fibrotic effects in cultivated C. sinensis. Methods: Nucleosides, sterols and polysaccharides were separated and purified from cultivated C. sinensis, and analyzed by high performance liquid chromatography, gas chromatography-mass spectrometry and chemical chromogenic methods, respectively. In high glucose-induced rat mesangial cell models, fibronectin and type 1 collagen were used as evaluation indicators. Results: There were 10 kinds of nucleosides and one sterol in cultivated C. sinensis. The contents of nucleosides, sterols and polysaccharides in the cultivated C. sinensis were close to 2%, 0.55% and 4.4%, respectively. Furthermore, nucleoside, sterol and polysaccharide components exhibited varying degrees of antifibrotic activity. The nucleoside components and sterol components inhibited the expression of extracellular matrix more effectively in the three main components. Conclusion: Cultivated C. sinensis remains the similar compounds with the wild C. sinensis, and nucleosides and sterols may be the main active substances that contribute to its anti fibrotic effects. The project of this study may provide valuable information on further optimization of more effective remedies with few side effects based on cultivated C. sinensis.

Rapid identification of Astragalus membranaceus processing with rice water based on intelligent color recognition and multi-source information fusion technology

2025/10/29 15:42:48

Objective: This study seeks to optimize the processing parameters for Astragalus membranaceus with rice water (AM-RW), establish quality evaluation standards, and develop a rapid multilayer perceptron (MLP) model for classification. This model facilitates accurate identification of AM-RW at various processing stages, providing a scientific reference for the quality assessment of traditional Chinese medicine products. Methods: Optimization of AM-RW was achieved using a single-factor test and Box-Behnken design response surface methodology to determine the optimal process parameters. The Watershed Algorithm was applied to segment images of AM tablets, and the numpy and pandas libraries were used to collect color data from these tablets. The study also explored the correlation between R, G, B, and L color values and calycosin-7-glucoside content. A rapid classification model based on MLP was developed, utilizing color values, hardness values, and calycosin-7-glucoside content of AM-RW with various processing degrees. Results: The study identified the optimal parameters for AM-RW as 20 mL of rice water, a frying temperature of 180 °C, and a frying time of 7 min. The average color values for the best-processed products fell within the normal distribution range: R value (94.83 ± 8.57), G value (96.1 ± 19.37), B value (36.84 ± 5.93), and L value (89.55 ± 12.24). The rapid identification model using MLP demonstrated high accuracy and reliability, achieving an accuracy rate of 94% in the classification process. Conclusions: The response surface method effectively optimizes the precise processing parameters of AM-RW. Furthermore, the MLP-based model can accurately classify AM-RW with varying degrees of processing, providing a valuable reference for the expedited identification of processing quality in traditional Chinese medicine products.

Effect of different drying degrees on content of various chemical components and in vitro bioactivity of Chrysanthemi Indici Flos

Tao?Wang?a?1 *, Shuting?Dong?a?b?1, Qiaosheng?Guo?a, Qingjun?Zou?a, Feng?Yang?a — 2025/10/29 15:42:48

Objective: Chrysanthemi Indici Flos (CIF), the inflorescence of Chrysanthemum indicum L., is widely used in traditional Chinese medicine. Traditionally, the content of active components in Chinese medicinal materials often changes during processing. However, the variation patterns of main active components in CIF during drying remain largely unknown. This study aims to investigate the contents of multiple chemical components and in vitro bioactivity of CIF with different drying degrees, so as to provide an optimized scheme for its origin processing. Methods: Fresh CIF were dried at 60 °C to obtain samples with different water contents (80%WC, 60%WC, 40%WC, 20%WC and DS). HPLC was used to determine eight chemical components (chlorogenic acid, 3,5-O-dicaffeoylquinic acid, galuteolin, linarin, luteolin, naringenin, kaempferol and isorhamnetin). In vitro assays evaluated antioxidant activity (radical scavenging), anti-inflammatory activity (enzyme inhibition) and antibacterial activity (disk diffusion, MIC) against several strains. Results: Among eight chemical components, chlorogenic acid and linarin decreased significantly with drying, luteolin and kaempferol first increased then decreased (peaking at 40%WC), and naringenin was highest in 80%WC. HPLC fingerprint similarity was lowest (0.559) between 80%WC and DS samples. Antioxidant (Total reducing capacity/DPPH/ABTS/·OH IC50: 80%WC > DS) and anti-inflammatory (XOD/LOX IC50: 80%WC > DS) activities strengthened with drying. Extracts inhibited Staphylococcus aureus (MIC 1.560–3.125 mg/mL), Listeria monocytogenes (DS group inhibition zone larger) and Escherichia coli (MIC 3.125 mg/mL), but not Pseudomonas aeruginosa or Salmonella paratyphoid. Conclusion: Drying degree significantly affects CIF’s chemical profiles and in vitro bioactivities, with dry samples showing superior antioxidant/anti-inflammatory properties and specific antibacterial effects. These findings offer targeted guidance for optimizing primary processing parameters of CIF.

Rhein alleviates renal interstitial fibrosis by inhibiting Smad3 phosphorylation in TGF-β/Smad signalling pathway

2025/10/29 15:42:48

Objective: The anthraquinone compound rhein (1,8-dihydroxy-3-carboxyanthraquinone), derived from Rhei Radix et Rhizoma (rhubarb, Dahuang in Chinese), exhibits notable anti-fibrotic effects. However, the mechanisms underlying these effects have not been fully elucidated. Suppressor of mothers against decapentaplegic 3 (Smad3) phosphorylation plays a crucial role in the canonical transforming growth factor-β (TGF-β)/Smad signalling pathway. In this study, we investigated the effect of rhein on the TGF-β/Smad signalling pathway in renal interstitial fibrosis (RIF). Methods: A unilateral ischaemia–reperfusion injury (UIRI) rat model was employed to simulate renal injury and assess the therapeutic effect of rhein in vivo. In vitro, TGF-β1-stimulated NRK-52E rat renal epithelial cells and HK-2 human proximal tubular epithelial cells were used to mimic fibrotic conditions. Rhein’s interaction with Smad3 was further explored using molecular docking and bio-layer interferometry assays. Additionally, Smad3 knockdown and overexpression studies were performed in HK-2 cells to elucidate the functional role of Smad3 in rhein-mediated anti-fibrotic activity. Results: Rhein treatment significantly improved renal function and reduced fibrosis in UIRI rats, primarily by inhibiting Smad3 phosphorylation. Rhein treatment mitigated aberrant remodelling and extracellular matrix accumulation in both NRK-52E and HK-2 cells and in the UIRI rat model. The anti-fibrotic effects of rhein were attenuated by Smad3 deficiency but enhanced by Smad3 overexpression in HK-2 cells. Conclusion: Rhein exerts its anti-fibrotic effects in renal interstitial fibrosis by targeting the TGF-β/Smad3 signaling pathway. Acting as a natural antagonist of Smad3, rhein offers promising potential for therapeutic development in renal fibrosis. These findings provide a new mechanistic insight for further clinical research and drug development.

Ganoderic acid a derivative induces apoptosis of cervical cancer cells by inhibiting JNK pathway

2025/10/29 15:42:48

Objective: Ganoderic acid A can inhibit the proliferation and promotes the apoptosis of cancer cells. Surprisingly, the molecular mechanisms underlying the anti-cancer effects of ganoderic acid A still remain poorly defined. Ganoderic acid A derivative (GaAD19) is an effective ingredient obtained by structural modification of ganoderic acid A. The purpose of this study was to evaluate the anti-proliferation effect of GaAD19 on cervical cancer cells. Methods: Through the HeLa cervical cancer cell model, the drug target of GaAD19 was predicted using the SwissTargetPrediction database and molecular docking. Subsequently, computer analysis results were verified by a series of molecular biology experiments, such as flow cytometry, Western blot, immunocytochemical staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), quantitative real time polymerase chain reaction (qPCR), and so on. Then, pathway agonists and inhibitors were used to investigate the mechanism of GaAD19. Finally, the mouse model of cervical cancer was established to evaluate the inhibitory effect of GaAD19 on tumor growth in U14 cervical cancer mice. Results: GaAD19 induced apoptosis and inhibited the growth of tumors. It also blocked the transition from the G1 to the S phase of the cell cycle. However, in the presence of a c-Jun N-terminal kinase (JNK)agonist, the effects of GaAD19 on the proliferation, apoptosis, and cell cycle transition of cancer cells were suppressed. Conclusion: This study showed that GaAD19 can play an anti-cervical cancer role by inhibiting the JNK signaling pathway. These results will be helpful in further exploring the mechanism of GaAD19 in the treatment of cervical cancer.

Salvia miltiorrhiza water extract ameliorated cGAS-STING-mediated acute liver injury by blocking interaction between STING and TBK1

2025/10/29 15:42:49

Objective: To investigate the potential of Salvia miltiorrhiza water extract (SMWE) as a modulator of the cGAS-STING signaling pathway, which is implicated in the pathogenesis of immune and inflammatory disorders, and to elucidate its underlying mechanism of action through in vitro and in vivo experiments. Methods: The cGAS-STING signaling pathway was activated in bone marrow-derived macrophages (BMDMs), Tohoku hospital pediatrics-1 (THP-1) cells, and peripheral blood mononuclear cells (PBMCs). The effect of SMWE on the expression of phosphorylated interferon regulatory factor 3 (IRF3) and phosphorylated STING after aberrant activation of the cGAS-STING pathway was detected by immunoblotting. Subsequently, real-time quantitative PCR was performed to detect changes in the mRNA levels of interferon type I (IFN), interferon-stimulated genes and inflammatory factors. The effect of SMWE on STING oligomerisation and the interaction between STING, Tank Binding Kinase 1 (TBK1) and IRF3 was investigated by immunoblotting. A model of acute liver injury (ALI) caused by lipopolysaccharide/D-galactosamine (LPS/D-GaIN) was used to test the effects of SMWE on inflammation mediated by the cGAS-STING signaling cascade. Results: SMWE significantly inhibited the phosphorylation of STING and IRF3, thereby suppressing the activation of the cGAS-STING signaling pathway. It also stopped the cGAS-STING pathway from working by stopping the production of type I interferons and interferon-stimulated genes, like interferon-stimulated gene 15 (ISG15) and C-X-C motif chemokine ligand 10 (CXCL10). SMWE also reduced the production of pro-inflammatory cytokines, such as interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α). SMWE also significantly improved ALI resulting from LPS/D-GaIN by diminishing the hyperactivation of the cGAS-STING signalling pathway. Mechanistic analysis revealed that SMWE disrupted the interaction between STING and TBK1. Conclusion: SMWE is a potent modulator of aberrant activation of the cGAS-STING pathway and is able to prevent and treat LPS/D-GaIN-induced ALI by inhibiting activation of the cGAS-STING pathway.

Human cytochrome P450 enzymes catalyze oxidative metabolism of pectolinarigenin to generate a more active Nrf2 agonist

2025/10/29 15:42:49

Objective: To characterize the oxidative metabolic pathway(s) of pectolinarigenin (PEC) and reveal the effect of PEC oxidative metabolism on its biological activities including peroxisome proliferator-activated receptors (PPAR) and nuclear factor erythroid 2-related factor 2 (Nrf2) agonist effects, as well as the anti-oxidative and hepatoprotective activities. Methods:The oxidative metabolites of PEC were identified by liquid chromatography-time of flight-mass spectrometry (LC-TOF-MS/MS). The key enzymes involved in oxidative metabolism of PEC were assigned by P450 reaction phenotyping assays and enzymatic kinetics assays. Luciferase reporter assays and western blotting analysis were used to evaluate the Nrf2 and PPAR agonist effects of PEC and its oxidative metabolites. The intracellular levels of total reactive oxygen species (ROS), mitochondrial membrane potential (MMP), lactate dehydrogenase (LDH) and glutathione (GSH) in acetaminophen (APAP)-challenged hepatocytes were also tested. Results:PEC could be readily metabolized to form two O-demethylated metabolites including hispidulin (HIS, 4′-O-demethylated PEC) and 6-O-demethylated PEC in human liver microsomes (HLM) in the presence of nicotinamide adenine dinucleotide phosphate (NADPH), while HIS was identified as the major oxidative metabolite of PEC. At least nine human cytochrome P450 enzymes (CYP) enzymes could catalyze PEC-4′-O-demethylation, while CYP1A2 and CYP2D6 showed the highest binding affinities and rapid metabolic clearance rates in the oxidative metabolism of PEC. Biological assays showed that PEC 4′-O-demethylation slightly decreased the PPAR agonist effects of PEC, while HIS showed more potent Nrf2 agonist effect. Compared with PEC, HIS showed more efficacious hepatoprotective effect against APAP-induced hepatocyte injury, evidenced by more potent ability to reduce intracellular ROS and LDH levels, as well as more effective ability to elevate the intracellular levels of both MPP and GSH in APAP-challenged hepatocytes. Conclusion:CYPs catalyze PEC-4′-O-demethylation to generate a more active Nrf2 agonist (HIS), which shows more efficacious hepatoprotective effects against APAP-induced hepatocyte injury.

Jiaotaiwan activates serum SCFAs and upregulates cAMP-PKA-CREB-BDNF signaling pathway for antidepressant effects: A multicenter, randomized, controlled study

2025/10/29 15:42:49

Objective:Jiaotaiwan (JTW) is a classic traditional Chinese medicine (TCM) prescription for treating depression, but its potential mechanisms are not fully understood. The aim of this study is to detect the levels of serum Short-chain fatty acids (SCFAs) and cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA)-cAMP-response element binding protein (CREB)-brain derived neurotrophic factor (BDNF) signaling pathway, further revealing the scientific connotation of the antidepressant effect of JTW. Methods:In this multicenter, randomized, controlled study, 120 patients with depression were divided into the JTW (16.5 g/d) group, JTW (16.5 g/d) + selective serotonin reuptake inhibitors (SSRIs) group, and SSRIs group. Hamilton depression Scale-24 (HAMD-24) and Self-rating depression scale (SDS) were used for efficacy evaluation. Enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) were used to evaluate the expression levels of cAMP-PKA-CREB-BDNF signaling pathway. Serum SCFAs concentrations were analyzed using liquid chromatograph-mass spectrometer (LC-MS) targeted metabolomics. Results:After eight weeks of treatment, HAMD score and SDS score were significantly decreased in the three groups, and HAMD score in JTW + SSRIs group was significantly lower than that in SSRIs group. After treatment, the expression levels of cAMP-PKA-CREB-BDNF signaling pathway were significantly increased in the three group, with the JTW + SSRIs group showing more significant increase. After treatment, the levels of isobutyric, butyric, isovaleric, and valeric acids in the JTW + SSRIs groups were significantly higher than that before treatment, and the levels of isobutyric, and isovaleric acids in the JTW + SSRIs group was significantly higher than that in the JTW group and SSRIs groups. Conclusion:JTW can alleviate symptoms in patients with depression, and its antidepressant mechanism may be related to regulating serum SCFAs and cAMP-PKA-CREB-BDNF signaling pathway.

Microbiota basis of antidepressant effects of Zhi-zi-chi Decoction: Lactobacillus rhamnosus and gut microbiota metabolism

2025/10/29 0:00:00

Objective: Zhi-zi-chi Decoction (ZZCD) is a classical antidepressant traditional Chinese medicine (TCM) formula. However, the microbiological basis of its antidepressant effects remains unclear. The objective of this study was to investigate the microbial basis of ZZCD. Methods: The bacteria that exhibit enhanced growth in ZZCD extracts were identified through in vitro culture. A depressed rat model was subsequently established to assess the antidepressant effects of the bacteria. A GC–MS-based metabolomics approach and molecular docking simulation were employed to investigate the potential mechanism of the antidepressant effects of the bacteria from the perspective of changes in the metabolic profile of the gut microbiota. Finally, the antioxidant effects of gut microbial metabolites following Lactobacillus rhamnosus treatment were validated through cellular experimentation. Results: ZZCD was able to promote the growth of L. rhamnosus. Furthermore, L. rhamnosus was demonstrated to increase the percentage of sucrose preference, reduce the immobility time in tail suspension and forced swimming tests, and alleviate depressive symptoms in rats. Additionally, L. rhamnosus was shown to alter the metabolic profile of gut microbiota. A total of six metabolites (ethanamin, butyric acid, 2-methylbutanoic acid, behenic acid, 3-methylvaleric acid and 1-hexadecanol) associated with depressive behavior in rats were identified. All six metabolites demonstrated the capacity to bind to kelch-like ECH-associated protein 1 (Keap 1) in molecular docking simulations. Gut microbial metabolites after L. rhamnosus treatment increased nuclear factor erythroid 2-related factor 2 (Nrf2) levels in HT22 cells. Conclusion: L. rhamnosus may be one of the microbial bases for the antidepressant effects of ZZCD. Its effects may be related to the regulation of gut microbiota metabolism and anti-oxidative stress.

Exploring new therapeutic potential of five commonly used Pteris medicinal plants through pharmaphylogenomics and network pharmacology

2025/10/29 15:42:49

Objective: This study investigates the novel medicinal potential of five commonly used Pteris genus plants, including Pteris cretica, P. multifida, P. wallichiana, P. semipinnata, and P. vittata, by analyzing their chloroplast genomes, chemical compositions, and pharmacological activities. Methods: We conducted a rigorous phylogenetic analysis of these five pteridophytes to elucidate their evolutionary relationships. Chemical constituents were subjected to network pharmacology analysis to investigate their potential medicinal properties. Protein–protein interaction (PPI) networks and compounds-disease target networks were constructed using Cytoscape 3.7.2. Functional annotations, including Gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, were conducted by the Metascape platform. Finally, molecular docking simulations were carried out with AutoDock software to validate key findings. Results: Our study revealed a close phylogenetic relationship between P. cretica and P. multifida, with P. cretica exhibiting a higher enrichment score for COVID-19-related pathways. Network pharmacology analysis of P. cretica identified 21 compounds and 540 targets, including 69 associated with COVID-19. The potential therapeutic effects of P. cretica were linked to virus infection, inflammation, and immune response modulation. Core pathways included “coronavirus disease-COVID-19”, “JAK-STAT signaling pathway”, and “Toll-like receptor signaling pathway”. Molecular docking identified (22E)-5α,8α-epidioxyergosta-6,22-dien-3β-ol, notholaenic acid, 2β,15α-diol-ent-kaur-16-ene, and multikaurane A as vital compounds for COVID-19 treatment in P. cretica. Conclusion: This study highlights P. cretica as a potential candidate for COVID-19 prevention and treatment, identifying its key active components and mechanisms. These findings provide a scientific basis for further development of potential therapeutic agents based on P. cretica.

Unveiling bioactive components in Sargassum fusiforme via phytochemical and biological evaluation: A heterogeneous information ensemble approach

2025/10/29 15:42:49

Objective: Sargassum fusiforme (SF), a brown algae, is a marine traditional Chinese medicine with nutritional value. However, studies on the bioactive components in SF are limited. The aim of this study is to investigate phytochemical and biological properties of SF from different geographical origins and botanical parts, as well as to explore bioactive components within SF. Methods: SF from different origins and botanical parts were collected for integrated phytochemical analyses using ultra-high performance liquid chromatography-tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS), mass spectral molecular networking and liquid chromatography-single quadrupole mass spectrometry (LC-MSD), etc., followed by the systematic chemical profile evaluation. Subsequently, the glycolipid metabolism regulation and anticoagulation of SF were examined. Furthermore, the overall information acquired was assembled using multivariate analysis and machine learning to discover bioactive components. Results: A total of 123 compounds in SF were identified with the assistance of mass spectral molecular networking. The variations in the content of amino acids, fucoidan, alginate, phlorotannin, alkaloids, terpenoids, etc. were observed in SF from different origins and botanical parts. SF possessed the hypolipidemic, hypoglycemic and anticoagulant effects, and SF from Dongtou, Wenzhou, Zhejiang, was outstanding among origins in both phytochemical and biological evaluations. A heterogeneous information ensemble approach was adopted to discover the bioactive components in SF, namely 3-indoleacrylic acid, phlorotannin and fucoidan. Conclusion: This study evaluated the phytochemical and biological properties of SF, revealed the bioactive components using heterogeneous information and provided a reference for the rational consumption and further development of SF.

Exploration of chemical components and Rac1-dependent anti-hepatic fibrosis mechanism by total flavonoids derived from Tetrastigma hemsleyanum

Xin?Wu?a?1, Cuiwei?Chen?b?1, Hongyan?Zhang?b, Keda?Lu?c *, Gang?Cao?a * — 2025/10/29 15:42:49

Objective: This study is mainly to elucidate the characteristic components, hepatoprotective effect, and the underlying anti-hepatic fibrosis molecular mechanism of the total flavonoids of Tetrastigma hemsleyanum Diels & Gilg (SYQTF). Methods: The components of SYQTF were confirmed based on global natural products social molecular networking (GNPS) as well as the hepatoprotective effect on CCl4-induced acute liver injury. Cell toxicity and viability, cell cycle, and apoptosis analyses were then performed to determine the effects of SYQTF in transforming growth factor-β1 (TGF-β1) (10 ng/mL)-induced HSC-T6 hepatic fibrosis cell model. Finally, the lentiviral particles to package recombinant Ras-related C3 botulinum toxin substrate 1 (Rac1) over-expression and Rac1 knockdown plasmids were transfected into TGF-β1-induced HSC-T6 cells to elucidate the mechanism of anti-hepatic fibrosis of SYQTF targeting Rac1, respectively. Results: Our findings demonstrated that 86 components in SYQTF were identified in MSE and FastDDA mode with the aid of GNPS. Moreover, S15-ZJ, the genuine functional food and medicinal materials of T. hemsleyanum, was significantly different from other regions by multivariate statistical methods based on progenesis QI. Remarkably, kaempferol-3-O-furananose7-O-rhamnosyl-glucoside, the highest response intensity in S15-ZJ, may act as the potential Q-marker of genuine functional food and medicinal materials of T. hemsleyanum in Zhejiang Province distinguished from other areas. In addition, the in vivo experiments revealed that SYQTF greatly alleviated CCl4-induced acute liver injury. Furthermore, SYQTF can reversible the activation and proliferation of HSC-T6 cells, block the cell cycle in the G0/G1 phase, and promote cell apoptosis. Remarkably, the study is the first to reveal that SYQTF exhibited the anti-hepatic fibrosis effect through targeting Rac1 in the hedgehog signaling pathway. Conclusion: Rac1 is an important signaling molecule for the activation of the hedgehog pathway in HSC-T6. Importantly, SYQTF exerted its anti-hepatic fibrosis effect by inhibiting the Rac1-hedgehog signaling pathway. Furthermore, kaempferol-3-O-furananose7-O-rhamnosyl-glucoside could be a potential Q-marker of T. hemsleyanum in Zhejiang Province distinguished from other areas.

Anthraquinones of Cassiae Semen alleviate lipid accumulation in obesity by regulating brown adipose tissue and liver function

2025/8/4 10:57:12

Objective: Cassiae Semen (CS, Juemingzi in Chinese) is a widely used traditional Chinese medicine with a variety of pharmacological effects. This study aimed to investigate the potential therapeutic effects and molecular mechanisms of anthraquinones of CS (AQS) for adiposity. Methods: The chemical components of the AQS were determined using high-performance liquid chromatography (HPLC). Network pharmacology analysis was used to predict potential anti-obesity targets of action for AQS. We constructed high fat with high sugar water diet-induced obese mice and observed their body weight and whole-body lipid metabolism to evaluate the efficacy of AQS in promoting lipid metabolism. Subsequently, the epidermal temperature at the brown adipose tissue (BAT) before and after cold stimulation was observed and the expression of lipid metabolism-related genes in the liver and BAT tissues was detected to clarify the mechanism of action of AQS. Results: Network pharmacology analysis showed that AQS was involved in the regulation of liver and adipose tissue function under obesity. Pathological and biochemical results showed that AQS reduced lipid accumulation in the liver and adipose tissue induced by an unhealthy diet. With the increase of cold tolerance, the volume and weight of BAT were increased by AQS, suggesting that it regulated the body heat production dominated by BAT. After AQS treatment, the levels of genes related to uncoupling protein1 (UCP1) mediated adaptive thermogenesis in BAT tissues and lipid metabolism in the liver were also increased, which further proved that AQS activated BAT function to promote lipid metabolism in the whole body. Conclusion: This study revealed the pharmacological effects of AQS, thereby providing a scientific basis for regulating BAT thermogenesis and liver lipid metabolism to alleviate obesity and providing clues for further exploring the application of natural active ingredients in the treatment of metabolismrelated diseases.

Qingjie Fuzheng Granule prevents colitis-associated colorectal cancer by inhibiting abnormal activation of NOD2/NF-κB signaling pathway mediated by gut microbiota disorder

2025/8/4 10:57:12

Objective: This study investigates the efficacy and mechanisms of Qingjie Fuzheng Granules (QFG) in inhibiting colitis-associated colorectal cancer (CAC) development via RNA sequencing (RNA-seq) and 16S ribosomal RNA (rRNA) correlation analysis. Methods: CAC was induced in BALB/c mice using azoxymethane (AOM) and dextran sulfate sodium (DSS), and QFG was administered orally to the treatment group. The effects of QFG on CAC were evaluated using disease index, histology, and serum T-cell ratios. RNA-seq and 16S rRNA analysis assessed the transcriptome and microbiome change. Key pharmacodynamic pathways were identified by integrating these data and confirmed via Western blotting and immunofluorescence. The link between microbiota and CACrelated markers was explored using linear discriminant analysis effect size and Spearman correlation analysis. Results: Long-term treatment with QFG prevented AOM/DSS-induced CAC formation, reduced levels of interleukin (IL)-1b, tumor necrosis factor-alpha (TNF-a), IL-6, and interferon c (IFN-c), and increased CD3+ and CD4+/CD8+ T cells ratio, without causing hepatic or renal toxicity. A 16S rRNA analysis revealed that QFG rebalanced the Firmicutes/Bacteroidetes ratio and mitigated AOM/DSS-induced microbiota disturbances. Transcriptomics and Western blotting analysis identified the nucleotide-binding oligomerization domain-containing protein 2 (NOD2)/nuclear factor kappa-B (NF-jB) pathway as key for QFG’s treatment against CAC. Furthermore, QFG decreased the abundance of Bacilli, Bacillales, Staphylococcaceae, Staphylococcus, Lactobacillales, Aerococcus, Alloprevotella, and Akkermansia, while increasing Clostridiales, Lachnospiraceae, Lachnospiraceae_NK4A136_group, Ruminococcaceae, and Muribaculaceae, which were highly correlated with CAC-related markers or NOD2/NF-jB pathway. Conclusion: By mapping the relationships between CAC, immune responses, microbiota, and key pathways, this study clarifies the mechanism of QFG in inhibiting CAC, highlighting its potential for clinical use as preventive therapy.

Riboflavin metabolism involved into comparison of therapeutic action of wild-simulated and transplanted Astragali Radix to treat chronic atrophic gastritis rats based on endogenous-xenobiotics metabolism

2025/8/4 10:57:12

Objective: Astragali Radix (AR, Huangqi in Chinese) has been utilized generally as a bulk drug for the treatment of chronic atrophic gastritis (CAG) in China. The efficacy of wild-simulated AR (WAR) and transplanted AR (TAR) commercially available is unclear. This study aimed to further clarify the therapeutic action of WAR and TAR to treat CAG rats based on endogenous-xenobiotics metabolism. Methods: Ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometer (UHPLC-Q-TOF/MS) based metabolomics had been used to analyze the cecal contents metabolic features and metabolic process differences of two ARs in the treatment of CAG. Results: Apparent pharmacodynamic indicator examination revealed that the WAR group had a more substantial curative effect. Metabolomics studies revealed that seven endogenous metabolites were significant differences in two ARs. Furthermore, following treatment, 77 and 65 xenobiotics metabolites were identified in the WAR and TAR groups, respectively. Finally, the most critical riboflavin metabolic route in the formation of CAG was chosen for molecular docking with the relevant exogenous components, and WAR scored higher than TAR. Conclusion: In this work, we analyzed the efficacy differences of AR from diverse cultivation forms by combining metabolomics and medicinal chemistry technologies, and it gave a fresh perspective for TCM efficacy evaluation and quality control research.

Coptis chinensis shows distinct effects on hyperlipidemia and gut microbiota in high-fat diet induced mice with cold or hot syndrome

Yanan Yang a, Jiaguo Zhan a, Jiale Cheng a, Ying Cao a, Chongming Wu a,b,? — 2025/8/4 10:57:12

Objective: Coptis chinensis (Huanglian in Chinese, HL) is commonly utilized in clinical settings to counteract dyslipidemia in patients with hot syndrome. Its lipid-reducing efficacy has been consistently demonstrated in high-fat diet (HFD)-induced hyperlipidemic animal models. However, whether HL’s efficacy differs in HFD-fed animals with hot or cold syndromes remains unclear. This study aims to discern the variations in the anti-hyperlipidemic effects of HL in HFD-fed mice with hot or cold syndromes. Methods: HFD-induced C57BL/6 mice were subjected to cold or hot syndrome via two weeks of ice water (0 °C) and levothyroxine sodium (240 lg/kg) treatment, respectively. Then, an aqueous extract of HL was administered to the mice via oral gavage over the following four-week period. Lipid levels in the serum and liver were gauged to determine the lipid-reducing effects of HL. Furthermore, gut microbiota composition was elucidated using full-length 16S rRNA gene sequencing. Results: HL notably reduced lipid levels in HFD-induced hyperlipidemic mice. Its efficacy was amplified in hyperlipidemic mice with a hot syndrome but was markedly reduced in those with a cold syndrome. HL treatment led to a decline in alpha-diversity (characterized by ACE, Chao1, Shannon and Simpson index) of the gut microbiota in both sets of mice but affected specific microbial populations based on the syndrome. Specifically, while HL led to a notable increase in Eubacterium, Robinsoniella, and Lachnoclostridium genera, along with the enhancement of Clostridium innocuum and Bacteroides thetaiotaomicron species across all conditions, it syndrome-dependently stimulated Romboutsia ilealis and Parabaceroides_sp_HGS0025 species in mice with hot syndrome. Conclusion: HL shows stronger lipid-lowering effect on hyperlipidemic mice with hot syndrome, which is in accordance with its traditional usage in clinic. The therapeutic outcomes of HL are intrinsically tied, at least in part, to its modulatory effects on the gut microbiota, offering fresh insights into the foundational principles of traditional Chinese medicine.

Hyssopus cuspidatus extract inhibited OVA-sensitized allergic asthma through PI3K/JNK/P38 signaling pathway and lipid homeostasis regulation

2025/8/4 10:57:12

Objective: To investigate the effect and mechanism of Hyssopus cuspidatus Boriss. extract (HCE) in ovalbumin (OVA)-induced allergic asthma. Methods: Components identification of HCE was conducted using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry. Mice were sensitized with OVA to establish asthmatic model, and dexamethasone was used as positive control. Respiratory reactivity, white cells counting in bronchoalveolar lavage fluid and peripheral blood, cytokine level measurement in serum and lung tissue, and histologic examination were performed to evaluate the therapeutic effect of HCE on asthma. Network pharmacology approach was used for mechanism prediction. Western blotting and untargeted lipidomics method were applied for mechanism validation. Results: Fifty-two compounds were identified in HCE, predominantly terpenoids and flavonoids. HCE markedly reduced airway resistance, the eosinophil infiltration in lung tissues, and the levels of immunoglobulin E, interleukin-4, interleukin-5, and interleukin-13. Network pharmacology analysis suggested phosphatidylinositol 3-kinases (PI3K), c-Jun N-terminal kinase (JNK), and p38 Mitogen-activated protein kinase (p38 MAPK) may be key proteins of HCE in the treatment of allergic asthma. Western blot results indicated that the levels of phosphorylated PI3K, JNK, and P38 were downregulated in HCEtreated group. Moreover, HCE significantly upregulated the levels of ceramide and sphingomyelin and downregulated the level of phosphatidylcholine. Conclusion: HCE inhibited allergic asthma via PI3K/JNK/P38 signaling pathway and lipid homeostasis regulation.

Compound Danshen Tablets ameliorate myocardial ischemia/reperfusion injury-induced ventricular remodeling by regulating autophagy via AMPK/mTOR signaling pathway

2025/8/4 10:57:12

Objective: Left ventricular remodeling induced by myocardial ischemia/reperfusion injury (MI/RI) is a common cardiac dysfunction. Accumulating evidence has demonstrated that autophagy plays a vital role in protecting against ventricular remodeling. This study aims to investigate the performance of Compound Danshen Tablets (CDT) in rescuing ventricular remodeling and whether autophagy as the potential mechanism. Methods: The left anterior descending arteries of rats were temporarily ligated for 30 min to construct the MI/RI model. Ventricular remodeling was induced by reperfusion for 28 d, during which the MI/RI rats were administered CDT (300 mg/kg and 600 mg/kg), atorvastatin (2 mg/kg), and diltiazem (16 mg/kg). Cardiac function and structure were examined by echocardiography. Immunohistochemistry, Masson’s trichrome staining, and hematoxylin-eosin (HE) staining were utilized to assess the fibrosis and histological alterations in the heart tissue. The expression of autophagy-related proteins was detected using Western blotting. Results: CDT attenuated the cardiac dysfunction, structural changes, histopathological changes and fibrosis induced by MI/RI. CDT significantly enhanced the level of Beclin1 and microtubule-associated protein 1 light chain 3 beta (LC3b), and reduced p62 levels in MI/RI rats. Moreover, CDT significantly increased the phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) and inhibited mammalian target of rapamycin (mTOR) phosphorylation. Conclusion: CDT ameliorated MI/RI-induced ventricular remodeling by activating autophagy and improving autophagic flux via the AMPK/mTOR signaling pathway.

Applications and challenges of DNA barcoding in rapid radiation groups: Rhodiola (Crassulaceae) as a case study

2025/8/4 10:57:12

Objective: Rhodiolae Crenulatae Radix et Rhizoma (Hongjingtian in Chinese, RCRR), the roots and rhizomes of Rhodiola crenulata and its application in the medicinal market is very chaotic. In this study, DNA barcoding database and identification engine of Rhodiola species were established, decoction pieces from the medicinal market were identified, and the application and challenges of DNA barcoding in the rapid radiation of Rhodiola species were analyzed. This study provides reference for the protection, rational development, and utilization of endangered resources within Rhodiola species. Methods: A total of 50 original plant samples from 20 species of the genus Rhodiola from Hebei, Xinjiang, Tibet, Jilin, and other major production areas were collected. Theses samples cover the typical distribution area (Qinghai-Tibetan Platea) of Rhodiola species and other scattered alpine regions (Changbai Mountain, Taibai Mountain, Lushan Mountain, etc.), it encompasses all Rhodiola species with thick rhizomes in China. ITS2 and psbA-trnH barcode of Rhodiola database (BORD) were established and an identification engine named Rhodiola-IDE was developed. The stability and accuracy of the standard DNA barcoding database were evaluated using two datasets. Rhodiola-IDE identified 31 decoction pieces of RCRR from the medicinal material market. Results: The BORD containing 1 532 sequences of 88 Rhodiola species has been established, and the identification efficiency results showed good accuracy and stability. According to the Chinese Pharmacopoeia (2020 edition), 23 samples (74.2%) were identified as authentic R. crenulata, while the rest of the marketed varieties were R. kirilowii, R. dumulosa, and R. fastigiata. The product label ‘‘Larger flower, Hongjingtian” was identified as R. crenulata. Samples labeled as ‘‘Smaller flower, Hongjingtian” were identified as R. crenulata, R. kirilowii, and R. fastigiata. Conclusion: ITS2 and psbA-trnH barcodes can identify monophyletic groups represented by R. crenulata. However, for non-monophyletic species, it is necessary to collect as many samples as possible and combine them with multiple markers for joint identification. This study discussed the application and challenges of DNA barcodes in Rhodiola under rapid radiation conditions, providing a scientific basis for the rational development and utilization of Rhodiola varieties.

Ecological factors impacting genetic characteristics and metabolite accumulations of Gastrodia elata

2025/8/4 10:57:12

Objective: The investigation of the correlation between ecological factors and the genetic characteristics or metabolites of plants offers valuable insights into the regional causes of genetic and metabolic diversity. Here, Gastrodia elata, a medicinal plant, is employed as a model to explore the environmental factors that influence its genetic characteristics and metabolic accumulations. Methods: A total of 23 G. elata populations from six cultispecies and 11 cultivated regions were selected based on the predictions of the global geographic information system. The genetic characteristics of these populations were evaluated using highly polymorphic simple sequence repeat markers. Additionally, the metabolic accumulations and antioxidant capacity of mature tubers were measured employing colorimetry and high performance liquid chromatography (HPLC). Ecological data of each region were obtained from the WorldClim-global climate database and harmonized world soil database. To assess the influence of ecological factors on the genetic characteristics and metabolic profiles of G. elata, Pearson’s correlation analysis was conducted. Results: Genetic variation among G. elata populations exceeded that within populations. Genetic diverisity, distance and structure manifested regional and species-specific patterns. Metabolic profiling and antioxidant capacity exhibited regional variations. Notably, the Lueyang region demonstrated that a content range of total polysaccharide, total protein, and phenolic glycosides was 9.34%-189.67% higher than the average. Similarly, in the Hubei region, total phenolic content, p-hydroxybenzyl alcohol content, and antioxidant indicators were observed to be higher than the average levels, by 106.57%, 136.47% and 12.50%-91.14%, respectively. Furthermore, ecological factors had a significant comprehensive impact on G. elata genetic characteristics (r > 0.256 and P < 0.05). Multivariate metabolite accumulations in G. elata were influenced by dominant ecological factors. Temperature notably impacted the accumulation of total protein (|r| > 0.528 and P < 0.05). Moisture, encompassing precipitation and soil content, significantly affected the production of phenolic glycosides (|r| > 0.503 and P < 0.05). Conclusion: The genetic characteristics of G. elata manifested regional and species-specific patterns, with the metabolic accumulations and antioxidant capacity of mature tubers exhibited regional variations. Specifically, multivariate ecological factors comprehensively influenced genetic characteristics. Temperature and moisture played pivotal roles in regulating the accumulations of proteins and phenolic glycosides, respectively. These findings underscore the significant impact of ecological factors on the shaping of G. elata, highlighting their crucial role in enhancing the quality of Chinese medicinal materials.

Research on comprehensive quality consistency evaluation strategy for TCM Granules: A case study with sugar-free Yangwei Granules produced by fluid-bed granulation

Jie Zhao, Geng Tian, Haibin Qu * — 2025/8/4 10:57:12

Objective: To develop a quality consistency evaluation strategy for traditional Chinese medicine (TCM) granules using sugar free Yangwei Granules as a model drug, and demonstrate the effectiveness of the developed method. Methods: The strategy integrates several methods including, HPLC fingerprint and physical fingerprint methods analyze the similarity in chemical and physical properties of the TCM granule samples. Nearinfrared (NIR) spectroscopy with principal components cluster analysis method is used to monitor normal operating conditions (NOC) samples accurately and to identify different types of abnormal operating conditions (AOC) samples, particularly those that deviate from the normal range. Results: The combined use of HPLC fingerprint and physical fingerprint provides insights into the chemical and physical properties of the samples. NIR spectroscopy, combined with principal components cluster analysis, achieves high accuracy in monitoring NOC samples and identifying AOC samples without misjudgment. The approach proves useful as a complementary method in cases where HPLC fingerprint and physical fingerprint alone lack sufficient resolution. Conclusion: This study establishes the feasibility and utility of the integrated approach for assessing the quality consistency of TCM granules. The strategy shows a high degree of generalization and holds significant importance for enhancing the quality control processes of TCM granules.

Traditional Chinese medicine formulas alleviated acute pancreatitis via improvement of microcirculation: A systematic review and meta-analysis

2025/8/4 10:57:12

Objective: Microcirculatory disturbance is pathologically critical to acute pancreatitis (AP), which can be effectively alleviated by traditional Chinese medicine (TCM) formulas that activate blood flow. However, there has been no evidence-based research to date. Therefore, a well-designed systematic review and meta-analysis is necessary to elucidate the therapeutic transformative benefit of improving microcirculation during AP. This study aims to confirm the therapeutic efficacy of TCM formulas and explore the potential mechanisms underlying their effects on AP treatment. Methods: Studies from eight databases including Pubmed, Embase, Web of Science, Cochrane Library, CNKI, CBM, Wanfang, and Chinese VIP, were screened for the eligible randomized controlled trials (RCTs). The APACHE II score and effectiveness rate were set as primary outcomes, while mortality rate, complications, total hospital stays, serum amylase recovery time, the time until the disappearance of abdominal pain, microcirculation indicators, and inflammation indicators were chosen as secondary outcomes. A systematic review and meta-analysis were subsequently conducted. Network pharmacology analysis was performed to analyze potential bioactive components with relevant targets of the core herbs included in the TCM formulas for activating blood flow. Results: A total of 51 RCTs (n = 3 721) were included. Compared with conventional western medical treatments alone, TCM groups were associated with lower APACHE II score (SMD = 1.36, 95% CI: 2.01 to 0.71, P = 0.000) and higher effectiveness rate (RR: 1.22, 95% CI: 1.18 to 1.26, P = 0.000). Furthermore, the formulas for activating blood flow demonstrated significant efficacy in improving both microcirculation and inflammation indicators. Additionally, six core Chinese herbal medicines including Rhei Radix et Rhizoma with the highest frequency, Aurantii Fructus Immaturus, Paeoniae Radix Rubra, Bupleuri Radix, Salviae Miltiorrhizae Radix et Rhizoma, and Corydalis Rhizoma were filtered out from the adopted TCM formulas. Finally, 166 shared targets between the six herbs and AP were identified. KEGG analysis indicated that lipid and atherosclerosis pathway is highly related to microcirculation. Conclusion: TCM formulas for activating blood flow significantly improve microcirculation and alleviate AP. Further high-quality, well-designed RCTs and deep mechanism exploration are required.

Shufeng Jiedu Granule against mild COVID-19: Protocol of the randomized, double-blind, placebo-controlled, multi-center heal-COVID phase III study

2025/8/4 10:57:12

Objective: Since Omicron will likely persist, this trial evaluates the safety and efficacy of Shufeng Jiedu Granule (SFJDG) for mild Omicron infection, aims at finding new therapies especially for home-treated patients. Methods: This randomized, double-blind, placebo-controlled, multi-center phase III trial involves 844 patients, divided into a treatment group (422) and control group (422). Participants will receive SFJDG or placebo for 7 d (1.2 g/bag, 2 bags, 3 times/d). Hospital evaluations will be done on days 1 and 8, with telephone assessments on days 3 and 5. Follow-up continues on days 10 and 14. Diary cards will track symptom scores and safety data. The primary outcome is the time to sustained clinical recovery from corona virus disease 2019 (COVID-19) symptoms. An interim analysis will occur after 70 % of patients complete follow-up, with Type I error correction (a1 = 0.015) at interim analysis based on O’Brien-Flemingtype cumulative error spending function. Results: This phase III trial evaluates the efficacy and safety of SFJDG for mild COVID-19, focusing on realworld applicability for home-managed patients. The study’s randomized, double-blind, placebocontrolled design ensures methodological rigor, while its comprehensive outcome measures address both symptom recovery and treatment safety. By emphasizing symptom resolution and recovery time, the trial aligns with the clinical priorities for managing mild cases of COVID-19. The findings could offer valuable insights into SFJDG’s role in improving patient outcomes and addressing gaps left by existing antiviral therapies, particularly in symptom management. Conclusion: The global risk assessment remains high due to the ongoing virulence of SARS-CoV-2 Omicron sub-lineages. This Phase III study adopts a robust methodology to investigate SFJDG as a treatment for mild COVID-19 as well as it’s effectiveness and safety. Furthermore, this study aim to provide sufficient scientific evidence for the market registration of SFJDG especially for home-treated patients. If successful, SFJDG could be a meaningful addition to therapeutic options for mild infections, supporting public health strategies in managing the ongoing impact of SARS-CoV-2.

MYB polymorphism molecular marker: A novel molecular marker for authenticity and geographical origin identification of Citri Reticulatae Pericarpium

2025/4/28 8:26:44

Objective: Citri Reticulatae Pericarpium (Chenpi, CRP) is one of the most used traditional Chinese medicines with great medicinal, dietary and collection values, among which the Citrus reticulata cv. ‘Chachi’ (Citrus reticulata cv. Chachiensis) from Guangdong Xinhui is the geoherb of CRP. Xinhui CRP in the market was often counterfeited with other varieties or origins, molecular identification method can effectively distinguish different CRP varieties, but it’s still a difficult problem to identify the same CRP variety from different origin. It is necessary to discover a newmolecular marker to ensure the safe and effective application of Xinhui CRP. Methods: We selected one of the most studied transcription factor families in Citrus genus, MYB, to design the specific candidate primers on the conserved region. The primers with good band repeatability and high polymorphism were screened for PCR amplification of the test materials, and the genetic similarity coefficient among different families, genera, species, and origins were calculated. The cluster analysis was performed by unweighted pair group method using arithmetic average (UPGMA). Results: A total of ten MYB primers were screened out to identify Xinhui CRP from plants from different family (Panax ginseng and Morus alba), genus (Clausena lansium and Zanthoxylum schinifolium), and species (Citrus reticulata, C. sinensis and C. maxima). Furthermore, two from the ten primers, M1 and M10, were found to distinguish Xinhui CRP from other origins. There were 169, 113, 133 and 134 polymorphic bands in the identification of different families, genera, species, and origins respectively, and the accordingly polymorphism ration were 79.88%, 76.87%, 79.20% and 82.84%. Moreover, M1was discovered to be the best primer to identify Xinhui CRP from other seven origins, the cluster analysis results based on the genetic similarity coefficients were consistent with the geographical distribution. Conclusion: This study established a novel molecular identification method according toMYB transcription factor, which can analyze the potential parental relationship of CRP germplasm, as well as identify the quality and origins of Xinhui CPR.

Identification and functional characterization of a new flavonoid glycosyltransferase from Rheum palmatum

2025/4/28 8:26:44

Objective: To characterize a glycosyltransferase (RpUGT1) from Rheum palmatum and investigate its specificity toward flavonoid compounds. Methods: The RpUGT1 was expressed in Escherichia coli and screened for catalytic activity against a range of flavonoid substrates using a high-throughput HPLC assay method. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) were used to determine the structure of the product. Homology modeling, molecular docking analyses and site-directed mutagenesis studies were conducted to identify key residues responsible for its function. Results: The recombinant RpUGT1 protein exhibited catalytic activity towards various flavonoids. Notably, RpUGT1 catalyzed the glycosylation of isorhamnetin to form 3-O-glucoside and kaempferol to form 7-O-glucoside, utilizing uridine diphosphate (UDP) glucose as the sugar donor. The homology modeling and molecular docking analyses identified key residues responsible for its activity. Subsequent sitedirected mutagenesis studies highlighted the crucial role of K307 in catalysis. Conclusion: These discoveries offer valuable perspectives on the role of the UGT family and establish a groundwork for forthcoming research on the synthesis of flavonoids in plants.

Analysis of fungal composition in different layers of Bantou agarwoodforming trunk of Aquilaria sinensis revealing presence of Aspergillusinhibiting substances in agarwood sites

Xuyu Chen a, Yun Yang a, Yangyang Liu a, Chun Sui b,?, Jianhe Wei a,b,? — 2025/4/28 8:26:44

Objective: The objective of this study was to analyse fungal composition and exploit application potential in the Bantou (BT) agarwood-forming trunk of Aquilaria sinensis. Methods: BT agarwood is a naturally formed agarwood that was collected after cutting. Total genomic DNA of the fungi in BT agarwood was extracted by the hexadecyltrimethy ammonium bromide (CTAB) method, followed by PCR amplification and library construction. The effective tags were obtained by the HiSeq2500 platform, and the data were subjected to bioinformatics and statistical analyses. Results: A total of 7 850 040 effective tags were obtained, Ascomycota was the most abundant fungus at the phylum level, with a relative abundance of 56.36%–61.44%, followed by Basidiomycota, with a relative abundance of 10.49%–20.39%. Dothideomycetes, Agaricomycetes and Sordariomycetes were dominant at the class level, accounting for 26.21%–33.88%, 8.40%–17.66%, and 18.41%–24.11%, respectively. Lignosphaeria, Phaeoacremonium and Hermatomyces were dominant at the genus level, with relative abundances of 6.25%–7.64%, 1.95%–9.05% and 1.5%–5.4%, respectively. Diversity and richness analysis showed that the fungal composition in the agarwood formation sites (agarwood layer, upper agarwood layer and lower agarwood layer) were significantly lower than those in the decomposing layer and the healthy layer. That is, the fungal diversity and richness were significantly reduced during agarwood formation by the action of open wounds. The fungal community structure in the decomposing layer and agarwood formation sites obviously differed from that in the healthy layer. The number of Aspergillus taxa in agarwood formation sites decreased significantly (healthy layer is 0.5%, decomposing layer is 0.022%, upper agarwood layer is 0.012%, agarwood layer is 0.01%, and lower agarwood layer is 0.013%), indicating that agarwood may contain potential substances to inhibit the growth of Aspergillus. Conclusion: Agarwood from agarwood formation sites contains potential substances that inhibit Aspergillus, which provides valuable information for the control of the genus of Aspergillus.

Enhancement of apoptosis in HCT116 and HepG2 cells by Coix lacryma-jobi var. lacryma-jobi seed extract in combination with sorafenib

2025/4/28 8:26:44

Objective: Coix lacryma-jobi, a highly regarded Asian herb widely used in traditional Chinese medicine, is recognized for its dual benefits in promoting overall health and treating various diseases. While it exhibits moderate anticancer efficacy when used alone, this study investigated the enhanced anticancer potential of raw and cooked Coix lacryma-jobi var. lacryma-jobi (CL) seed extracts in combination with sorafenib against HCT116 and HepG2 cancer cell lines. The combination of sorafenib with other anticancer agents, including natural extracts, has garnered significant attention as a promising strategy for developing more effective cancer therapies. Methods: Dry powders of raw (R) and cooked (C) CL seeds, obtained from a local commercial source in Thailand, were extracted and fractionated using ethanol (E), dichloromethane (D), ethyl acetate (A), and water (W) to produce eight fractions: CLRE, CLCE, CLRD, CLCD, CLRA, CLCA, CLRW, and CLCW. The coixol content in raw and cooked seed extracts was quantified and expressed as lg of coixol per gram of extract. The cytotoxic effects of these fractions were evaluated against HCT116 and HepG2 cells using the MTT assay. Fractions demonstrating the most significant cytotoxic responses were combined with sorafenib to evaluate their synergistic effects. Apoptosis induction and mitochondrial membrane potential (MMP) were assessed, and the underlying mechanism of apoptosis was explored by analyzing reactive oxygen species (ROS) generation and antioxidant protein expression levels. Additionally, the combination treatment’s effect on the phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) pathway was investigated. Results: One gram of CLCE and CLCD extracts contained higher coixol levels (7.02 lg and 9.69 lg, respectively) compared to CLRE and CLRD (2.66 lg and 5.96 lg, respectively). Coixol content in CLRA, CLRW, and CLCW fractions was undetectable under the study conditions. All extract fractions exhibited IC50 values exceeding 1 mg/mL after 24- and 48-hour incubations with HCT116 and HepG2 cells, indicating limited cytotoxicity when used independently. CLRD and CLCD fractions were selected for combination studies at a concentration of 1 mg/mL, combined with sub-IC50 concentrations of sorafenib to minimize its side effects. This combination significantly increased cytotoxicity, inducing apoptosis in HCT116 and HepG2 cells by elevating ROS levels and reducing the expression of superoxide dismutase 2 and catalase. Furthermore, the combination treatment downregulated the PI3K/AKT/mTOR pathway, indicating a targeted anticancer mechanism. Conclusion: The combination of CLCD with sorafenib demonstrates significant potential as a strategy for future anticancer therapies. This CL seed extract, cultivated and commercially available in Thailand, shows promise as a natural supplement to enhance the efficacy of chemotherapy in upcoming clinical anticancer applications.

Ameliorating vascular endothelial injury for lipolysacharide-induced via mitochondrial targeting function of octaarginine-modified essential oil from Fructus Alpiniae zerumbet (EOFAZ) lipid microspheres

2025/4/28 8:26:44

Objective: To investigate the therapeutic potential of octaarginine (R8)-modified essential oil from Fructus Alpiniae zerumbet (EOFAZ) lipid microspheres (EOFAZ@R8LM) for cardiovascular therapy. Methods: EOFAZ@R8LM was developed by leveraging the volatilization of EOFAZ and integrating it with the oil phase of LM, followed by surface modification with cell-penetrating peptide R8 to target the site of vascular endothelial injury. The therapeutic effects of this formulation in alleviating lipopolysaccharide-induced vascular endothelial inflammation were evaluated by assessing mitochondrial membrane potential (MMP), intracellular reactive oxygen species (ROS) levels, as well as inflammatory factors interleukin-6 (IL-6) and interleukin-1b (IL-1b) levels. Results: EOFAZ@R8LM effectively delivered EOFAZ to the site of injury and specifically targeted the mitochondria in vascular endothelial cells, thereby ameliorating mitochondrial dysfunction through regulation of MMP and reduction of intracellular ROS levels. Moreover, it attenuated the expression levels of IL-6 and IL-1b, exerting protective effects on the vascular endothelium. Conclusion: Our findings highlight the significant therapeutic potential of EOFAZ@R8LM in cardiovascular therapy, providing valuable insights for developing novel dosage forms utilizing EOFAZ for effective treatment against cardiovascular diseases.

Amelioration of atherosclerotic complications and dyslipidemia by verbascoside-enriched fraction of Clerodendrum glandulosum leaves targeting LDL-R and LXR-mediated reverse cholesterol transport

Puspanjali Khound a,b, Nonibala Gurumayum a,b, Rajlakshmi Devi a,b,? — 2025/4/28 8:26:44

Objective: Clerodendrum glandulosum is widely used in traditional Chinese and Indian systems of medicine for conditions like hypertension and diabetes. While various pharmacological benefits have been reported, research on its anti-atherosclerotic properties remains limited. Atherosclerosis (AS) is a chronic cardiovascular disease linked to dyslipidemia (DLD) and inflammation. This study aims to identify the bioactive fraction from C. glandulosum extract, evaluate its potential against AS and DLD, and explore the molecular mechanisms of cholesterol metabolism. Methods: Bioactivity-guided fractionation was employed to investigate the bioactivity of C. glandulosum by screening biochemical enzyme inhibitory potential. The active fraction was subjected to in vitro testing to assess the anti-inflammatory and anti-adhesion properties. The fraction was administered at 50 and 100 mg/kg per os (p.o.) to cholesterol-cholic acid-thiouracil (CCT) diet-induced atherosclerotic Wistar rats. Changes in lipid and antioxidant profiles, inflammatory markers, and cholesterol metabolism pathways were assessed using Western blotting. Histopathological analyses of the aorta, liver, heart, and kidneys were also conducted. Molecular docking was conducted for the verbascoside (VER) and the standard statin, atorvastatin (ATS), for their binding capabilities with the molecular targets considered in this study. Results: Bioactivity-guided fractionation and screening revealed that ethyl acetate fraction (EAF) contained VER as the principal phytoconstituent. EAF exhibited potent enzyme inhibitory activity, with IC50 values of 1.059 mg/mL for pancreatic lipase and 22.48 lg/mL for a-glucosidase. In vitro analysis revealed that EAF significantly lowered cell-to-cell adhesion to 0.57 folds from 2.5 folds in the disease control and normalized the inflammatory cytokines. In CCT-diet-induced rats, elevated serum cholesterol and low-density lipoproteins (LDL) levels (92.1 mg/dL and 78.49 mg/dL, respectively) were reduced to 63.52 mg/dL and 58.51 mg/dL with EAF at 100 mg/kg. EAF at 100 mg/kg reduced oxidized LDL to 53.63 ng/mL compared to 157.1 ng/mL in CCT-diet-fed rats. EAF also restored antioxidant activity by increasing superoxide dismutase and catalase levels to 73.78 and 17.72 U/mg protein, respectively, compared to 42.22 and 9.62 U/mg protein in CCT-diet-fed rats. EAF restored inflammatory cytokines to normal levels. Histological analyses validated the protective benefits of EAF supplementation for the structural integrity of the aorta, liver, heart, and kidney tissues. Western blotting analysis of liver tissues revealed changes in the cholesterol metabolic pathway by upregulating peroxisome proliferatoractivated receptor gamma (PPARc)/liver X receptor alpha (LXRa)/adenosine triphosphate-binding cassette sub-family G member 1 (ABCG1) and low-density lipoprotein receptor (LDL-R) expression. Unlike ATS, molecular docking analyses indicated strong interactions between VER and molecular targets. Conclusion: EAF prevented DLD and AS by reverse cholesterol transport via the PPARc/LXRa/ABCG1 pathway, offering potential therapeutic benefits for cardiovascular health.

Osthole ameliorates chronic pruritus in 2,4-dichloronitrobenzeneinduced atopic dermatitis by inhibiting IL-31 production

2025/4/28 8:26:44

Objective: This study aims to elucidate the therapeutic potential of osthole for the treatment of atopic dermatitis (AD), focusing on its ability to alleviate chronic pruritus (CP) and the underlying molecular mechanisms. Methods: In this study, we investigated the anti-inflammatory effects of osthole in both a 2,4- dichloronitrobenzene (DNCB)-induced AD mouse model and tumor necrosis factor-a (TNF-a) and interferon-c (IFN-c) stimulated human immortalized epidermal (HaCaT) cells. The anti-itch effect of osthole was specifically assessed in the AD mouse model. Using methods such as hematoxylin and eosin (HE) staining, enzyme-linked immunosorbent assay (ELISA), western blot (WB), quantitative real-time PCR (qRT-PCR), and immunofluorescence staining. Results: Osthole improved skin damage and clinical dermatitis scores, reduced scratching bouts, and decreased epidermal thickness AD-like mice. It also reduced the levels of interleukin (IL)-31 and IL-31 receptor A (IL-31 RA) in both skin tissues and HaCaT cells. Furthermore, Osthole suppressed the protein expression levels of phosphor-p65 (p-p65) and phosphor-inhibitor of nuclear factor kappa-Ba (p-IjBa). Meanwhile, it increased the protein expression levels of peroxisome proliferator-activated receptor a (PPARa) and PPARc in HaCaT cells. Conclusion: These findings indicated that osthole effectively inhibited CP in AD by activating PPARa, PPARc, repressing the NF-jB signaling pathway, as well as the expression of IL-31 and IL-31 RA.

Banxia Xiexin Decoction inhibits colitis-associated colorectal cancer development by modulating STAT3 signaling and gut microbiota

2025/4/28 8:26:44

Objective: To investigate the therapeutic effects of Banxia Xiexin Decoction (BXD), a herbal medicine formula, on inflammation and the imbalance of the gut microbiota in a rat model of colorectal cancer (CRC) induced by azoxymethane (AOM) /dextran sulfate sodium (DSS). Methods: A total of 75 male C57BL/6 mice were randomly divided into five groups: normal control group (NC), model group (MODEL), low-dose BXD treatment group (L-BXD), high-dose BXD treatment (H-BXD) group and MS treatment group (MS). BXD and MS were used in CRC mice at the doses of 3.915 g/kg, 15.66 g/kg, 0.6 g/kg for 3 weeks consecutively. Histopathological changes in the colon were observed using hematoxylin-eosin (HE) staining. The content of inflammatory factors in serum was detected by an enzyme-linked immunosorbent assay (ELISA), and the expression of mRNA and protein of genes related to immunity, apoptosis, inflammation, and inflammatory factors was evaluated. Changes in the intestinal flora of mouse fecal were determined based on high throughput sequencing of the 16S rRNA microbial gene. Results: Compared to the model group, the low-dose BXD and high-dose BXD groups decreased the number of colon tumors, reversed weight loss, and shortened colon length of mice. The pathological examination showed that BXD alleviated the malignancy of intestinal tumors. It also suppressed signal transducer and activator of transcription 3 (STAT3), matrix metalloproteinase-9 (MMP-9), and transforming growth factor beta 1 (TGF-b1) expression, while increasing the expression of the tight junction protein ZO-1 in colon tissues. Additionally, the levels of key pathway proteins involved in inflammation (phosphorylated-STAT3, Bcl-2, COX-2) and cell cycle regulatory molecules (c-Myc and PCNA) were reduced. According to 16S rRNA sequence analysis, BXD enhanced the relative abundance of potentially beneficial bacteria, while that of cancer-related bacteria decreased. Conclusion: BXD plays a preventive role in developing colorectal cancer; its mechanisms are related to the inhibition of inflammation and tumor proliferation, as well as maintenance of intestinal homeostasis.

Overexpression of SULT1E1 alleviates salt-processed Psoraleae Fructus-induced cholestatic liver damage

2025/4/28 8:26:44

Objective: Salt-processed Psoraleae Fructus (SPF) is widely used as a phytoestrogen-like agent in the treatment of osteoporosis. However, due to improper clinical use or misuse, resulting in liver damage. In this study, network pharmacology was employed to analyze the mechanism of cholestatic liver damage. An adeno-associated virus overexpressing SULT1E1 (rAAV8-SULT1E1) was constructed and the hepatotoxicity of SPF, psoralen, and isopsoralen was determined. Methods: By utilizing three databases inclding TCMSP, TCMID, and BATMAN- TCM, the targets of the three databases were summarized, and a total of 45 psoralen compounds were included. Network pharmacology analysis was then performed. The adenoviral vectors were injected into the tail vein of C57BL6 mice to elucidate the role of SULT1E1 in SPF-induced cholestasis-mediated hepatotoxicity in vivo. SPF (10 g/kg), psoralen, and isopsoralen (50 mg/kg each) were intragastrically administered to mice for 30 d. B-ultrasound and samples were collected and examined for follow-up experiments. Results: A total of 854 targets were predicted for 45 active components, with 151 cholestasis-mediated hepatotoxicity-related disease targets obtained for SPF. A total of 126 pathways were enriched based on KEGG pathway analysis, with the ‘‘estrogen signaling pathway” identified as one of the top 20 pathways. In terms of pathological hepatic changes, treated mice had visually swollen hepatocytes, dilated bile ducts, and elevated serum biochemical markers, which were more prominent in mice treated with isopsoralen than in those treated with other compounds. Notably, the overexpression of SULT1E1 could reverse liver damage in each treatment group. B-ultrasound was used to observe the size of the gallbladder in vivo. The size of the gallbladder was found to significantly increase on day 30 after treatment in the SPF-, psoralen-, and isopsoralen-treated groups, especially the SPF group. Compared with the expression levels in the negative control group (rAAV8-empty + con), the expression levels of FXR, Mrp2, Bsep, SULT1E1, SULT2A1, Ntcp, and Nrf2 decreased, whereas those of CYP7a1 and IL-6 increased in the SPF-, psoralen-, and isopsoralen-treated groups. Conclusion: The overexpression of SULT1E1 could alleviate the decreased or increased expression of indicators, indicating that SULT1E1 is an important target gene for SPF-induced liver damage. The severity of liver damage was significantly lower in the rAAV8 SULT1E1 groups than in the rAAV8-empty groups.

Two new polyketides from Rhodiola tibetica endophytic fungus Penicillium sp. HJT-A-6

2025/4/28 8:26:44

Objective: To study bioactive compounds from the endophytic fungus Penicillium sp. HJT-A-6 isolated from stem of Rhodiola tibetica, and evaluate its allelopathic activity. Methods: The chemical constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were elucidated by extensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. In addition, the allelopathic activity of compound 1 was evaluated by measuring the seed germination rate of R. tibetica. Results: Two new polyketides 4-hydroxy-3,6-dimethyl-2H-pyran-2-one (1) and penilactone E (2), together with six known compounds walterolactone A (3), 5-hydroxyhexan-4-olide (4), 3-methyl-2-penten-5-olide (5), chaetoquadrin F (6), (Z)-6-acetyl-3-(1,2 -dihydroxypropylidene)-5-hydroxy-8-methylchroman-2-one (7) and 4-hydroxy-3-(4-hydroxyhexanoyl)-5-methylfuran-2(5H)-one (8) were isolated from Penicillium sp. HJT-A-6. Compound 1 showed moderate seed-germination-promoting activity at a concentration of 0.001 mg/mL while inhibiting the seed germination at concentrations of 0.1 and 0.01 mg/mL. Compared with the positive drug 6-benzyladenine (6-BA), compound 1 could extend the seed-germination period of R. tibetica (up to 11 d). Conclusion: Two new compounds were isolated from R. tibetica endophytic fungus Penicillium sp. HJT-A-6. Compound 1 displayed plant hormone-like activity, which inhibited the seed germination of the host plant at high concentrations and promoted the seed germination of the host plant at low concentrations. The results not only enrich the chemical constituents of the endophytic fungi isolated from Rhodiola tibetica, but also provide a theoretical basis for understanding the interaction mechanism between Rhodiola tibetica endophytic fungi and the host plant.

TPMGD: A genomic database for the traditional medicines in Pakistan

2025/2/6 16:13:54

Objective: In Pakistan, traditional medicines are an important component of the medical system, with numerous varieties and great demands. However, due to the scattered resources and the lack of systematic collection and collation, adulteration of traditional Pakistani medicine (TPM) is common, which severely affects the safety of their medicinal use and the import and export trades. Therefore, it is urgent to systematically organize and unify the management of TPM and establish a set of standards and operable methods for the identification of TPM. Methods: We collected and organized the information on 128 TPMs with regard to their medicinal parts, efficacy, usage, and genetic material, based on Pakistan Hamdard Pharmacopoeia of Eastern Medicine: Pharmaceutical Codex. The genetic information of TPM is summarized from national center for biotechnology information (NCBI) and global pharmacopoeia genome database (GPGD). Furthermore, we utilized bioinformatics technology to supplement the chloroplast genome (cp-genome) data of 12 TPMs. To build the web server, we used the Linux + Apache + MySQL + PHP (LAMP) system and constructed the webpage on a PHP: Hypertext Preprocessor (PHP) model view controller (MVC) framework. Results: We constructed a new genomic database, the traditional Pakistani medicine genomic database (TPMGD). This database comprises five entries, namely homepage, medicinal species, species identification, basic local alignment search tool (BLAST), and download. Currently, TPMGD contains basic profiles of 128 TPMs and genetic information of 102 TPMs, including 140 cytochrome c oxidase subunit I (COI) sequences and 119 mitochondrial genome sequences from Bombyx mori, 1 396 internal transcribed spacer 2 (ITS2) sequences and 1 074 intergenic region (psbA-trnH) sequences specific to 92 and 83 plant species, respectively. Additionally, TPMGD includes 199 cp-genome sequences of 82 TPMs. Conclusion: TPMGD is a multifunctional database that integrates species description, functional information inquiry, genetic information storage, molecular identification of TPM, etc. The database not only provides convenience for TPM information queries but also establishes the scientific basis for the medication safety, species identification, and resource protection of TPM.

Paris saponin VII induces Caspase-3/GSDME-dependent pyroptosis in pancreatic ductal adenocarcinoma cells by activating ROS/Bax signaling

2025/2/6 16:13:54

Objective: Paridis Rhizoma (Chonglou in Chinese), a traditional Chinese herbal medicine, has been shown have strong anti-tumor effects. Paris saponin VII (PSVII), an active constituent isolated from Paridis Rhizoma, was demonstrated to significantly suppress the proliferation of BxPC-3 cells in our previous study. Here, we aimed to elucidate the anti-pancreatic ductal adenocarcinoma (PDAC) effect of PSVII and the underlying mechanism. Methods: Cell viability was determined by CCK-8, colony formation, and cell migration assays. Cell apoptosis and reactive oxygen species (ROS) production were measured by flow cytometry with annexin V/propidine iodide (Annexin V/PI) and 2',7'- dichlorodihydrofluorescein diacetate (DCFH-DA), respectively. Pyroptosis was evaluated by morphological features, Hoechst 33342/PI staining assay, and release of lactate dehydrogenase (LDH). JC-1 fluorescent dye was employed to measure mitochondrial membrane potential. Western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to determine the levels of proteins or mRNAs. The effect in vivo was assessed by a xenograft tumor model. Results: PSVII inhibited the viability of PDAC cells (BxPC-3, PANC-1, and Capan-2 cells) and induced gasdermin E (GSDME) cleavage, as well as the simultaneous cleavage of Caspase-3 and poly (ADP-ribose) polymerase 1 (PARP). Knockdown of GSDME shifted PSVII induced pyroptosis to apoptosis. Additionally, the effect of PSVII was significantly attenuated by Z-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethylketone (Z-DEVD-FMK), on the induction of GSDME-dependent pyroptosis. PSVII also elevated intracellular ROS accumulation and stimulated Bax and Caspase-3/GSDME to conduct pyroptosis in PDAC cells. The ROS scavenger N-acetyl cysteine (NAC) suppressed the release of LDH and inhibited Caspase-9, Caspase-3, and GSDME cleavage in PDAC cells, ultimately reversing PSVII-induced pyroptosis. Furthermore, in a xenograft tumor model, PSVII markedly suppressed the growth of PDAC tumors and induced pyroptosis. Conclusion: These results demonstrated that PSVII exerts therapeutic effects through Caspase-3/GSDMEdependent pyroptosis and may constitute a novel strategy for preventing chemotherapeutic resistance in patients with PDAC in the future.

Cerbera odollam fruit extracts enhance anti-cancer activity of sorafenib in HCT116 and HepG2 cells

2025/2/6 16:13:54

Objective: While higher therapeutic doses of toxic cardiac glycosides derived from Cerbera odollam are frequently employed in cases of suicide or homicide, ongoing research is investigating the potential anticancer properties of low-concentration extracts obtained from the fruits of C. odollam. The present study aimed to determine the enhanced anticancer effects and minimize potential side effects of combining extracts from C. odollam fruits from Thailand with sorafenib against HCT116 and HepG2 cells. Methods: The dried powder of fresh green fruits of C. odollam was fractionated, and its phytochemical contents, including total cardiac glycosides, phenolics, flavonoids, and triterpenoids, were quantified. The cytotoxic effects of these fractions were evaluated against HCT116 and HepG2 cells using the MTT assay. The fractions showing the most significant response in HCT116 and HepG2 cells were subsequently combined with sorafenib to examine their synergistic effects. Apoptosis induction, cell cycle progression, and mitochondrial membrane potential (MMP) were then assessed. The underlying mechanism of the apoptotic effect was further investigated by analyzing reactive oxygen species (ROS) generation and the expression levels of antioxidant proteins. Results: Phytochemical analysis showed that C. odollam-ethyl acetate fraction (COEtOAc) was rich in cardiac glycosides, phenolics, and flavonoids, while the dichloromethane fraction (CODCM) contained high levels of triterpenoids and saponins. Following 24 h treatment, HCT116 showed the most significant response to COEtOAc, while HepG2 responded well to CODCM with IC50 values of (42.04 ± 16.94) μg/mL and (123.75 ± 14.21) μg/mL, respectively. Consequently, COEtOAc (20 μg/mL) or CODCM (30 μg/mL), both administered at sub-IC50 concentrations, were combined with sorafenib at 6 μmol/L for HCT116 cells and 2 μmol/L for HepG2 cells, incubated for 24 h. This combination resulted in a significant suppression in cell viability by approximately 50%. The combination of treatments markedly enhanced apoptosis, diminished MMP, and triggered G0/G1 phase cell cycle arrest compared to the effects of each treatment administered individually. Concurrently, increased formation of ROS and decreased expression of the antioxidant enzymes superoxide dismutase 2 and catalase supported the proposed mechanism of apoptosis induction by the combination treatment. Importantly, the anticancer effect demonstrated a specific targeted action with a favorable safety profile, as evidenced by HFF-1 cells displaying IC50 values 2–3 times higher than those of the cancer cells. Conclusion: Utilizing sub-IC50 concentrations of COEtOAc or CODCM in combination with sorafenib can enhance targeted anticancer effects beyond those achieved with single-agent treatments, while mitigatin gopposing side effects. Future research will focus on extracting and characterizing active constituents, especially cardiac glycosides, to enhance the therapeutic potential of anticancer compounds derived from toxic plants.

Material basis revelation of anti-hepatoma effect of Huachansu (Cinobufacini) through down-regulation of thymidylate synthase

Qi Wu, Qimei Chen, Jingyi Yang, Jiayu Zhang *, Ailin Yang * — 2025/2/6 16:13:54

Objective: Hepatocellular carcinoma (HCC) is a leading cause of mortality worldwide. Huachansu (Cinobufacini) is active extract isolated from the dry skin of Bufo Bufo gargarizans. It has now been widely used in clinical treatment of cancer, this study is to clarify the material basis of down-regulation of thymidylate synthase (TYMS) induced by Huachansu. Methods: Our study utilized UPLC-MS/MS to identify major bioactive components from Huachansu. Cell Counting Kit 8 (CCK-8) assay and clone formation assay were used to examine the cell viability of tumor cells. TYMS and c-H2AX level were detected by using quantitative real-time RT-PCR and/or western blotting. Small interfering RNA (siRNA) transfection was used to explore whether inhibition of TYMS could enhance the suppressive effect of Huachansu on cell growth of HCC cells. Results: In our study, firstly, we identify 21 major bioactive components from Huachansu. CCK-8 assay results showed that Huachansu and its bioactive bufadienolides (Bufalin, Bufotalin, Cinobufotalin, Desacetylcinobufagin, Arenobufagin, Telocinobufagin, and Resibufogenin) significantly inhibited the proliferation of HepG2 and SK-HEP-1 cells in a dose- and time-dependentmanner. Further molecularmechanistic investigation demonstrates that Huachansu significantly suppresses thymidylate synthase (TYMS), the enzyme which provides the sole de novo source of thymidylate for DNA synthesis. The inhibition of TYMS could lead to cell-cycle block and DNA damage of HCC cells. Furthermore, we identified that Huachansu markedly increased c-H2AX expression, which indicated the presence of DNA damage. Moreover, we confirmed that transfection of cells with small interfering RNA specific to TYMS could increase the suppressive effects of Huachansu on the HCC cells proliferation. Quantitative RT-PCR analysis showed that Huachansu treatment had no effect on the transcription level ofTYMS. Furthermore, proteasomal inhibitorMG132 could blockTYMS inhibition induced by Huachansu, and concomitant administration of protein synthesis inhibitor cycloheximide (CHX) with Huachansu could further suppress the protein level of TYMS, indicating that Huachansu promotes proteasome-dependent degradation of TYMS in liver cancer cells. More importantly, the bioactive bufadienolides of Huachansu such as Bufalin, Bufotalin, Cinobufotalin, Desacetylcinobufagin, Arenobufagin, Telocinobufagin, and Resibufogenin could also significantly restrain the protein level of TYMS, revealing the material basis of inhibition of TYMS exposed to Huachansu. 5-Fluorouracil (5-FU) is a TYMS inhibitor, we also evaluate the effects of the combined treatment of Huachansu with 5-FU, the results show that interactions between Huachansu and 5-FU are synergistic or antagonistic. Thus, in clinical, attention should be paid to the dosage of Huachansu in combination with 5-FU. Conclusion: Huachansu inhibits the growth and induces DNA damage of human HCC cells through proteasome-dependent degradation of TYMS, bioactive bufadienolides are the material basis of downregulation of TYMS induced by Huachansu.

Qishen Granule protects against myocardial ischemia by promoting angiogenesis through BMP2-Dll4-Notch1 pathway

2025/2/6 16:13:54

Objective: Therapeutic angiogenesis has become a promising approach for treating ischemic heart disease (IHD). The present study aims to investigate the effects of Qishen Granule (QSG) on angiogenesis in myocardial ischemia (MI) and the potential mechanism. Methods: In vivo study was conducted on rat model of myocardial infarction. QSG was performed daily at a dose of 2.352 g/kg for four weeks. Cardiac function was assessed by echocardiogram and pro-angiogenic effects were evaluated by Laser Doppler and CD31 expression. Oxygen-glucose deprivation (OGD) was applied in cultured human umbilical vein endothelial cells (HUVECs). Cell viability, wound healing and tube formation assay were used to test functions of HUVECs. ELISA and Western blots were used to assess protein expressions of bone morphogenetic protein 2-delta-like 4-notch homolog 1 (BMP2-Dll4-Notch1) signaling pathway. Results: The results showed that QSG improved heart function, cardiac blood flow and microvessel density in myocardial ischemic rats. In vitro, QSG protected HUVECs by promoting the cell viability and tube formation. QSG upregulated bone morphogenetic protein-2 (BMP2) and downregulated delta-like 4 (Dll4) and notch homolog 1 (Notch1) expressions both in rats and HUVECs. Conclusion: QSG protected against MI by promoting angiogenesis through BMP2-Dll4-Notch1 pathway. BMP2 might be a promising therapeutic target for IHD.

Sini decoction alleviates inflammation injury after myocardial infarction through regulating arachidonic acid metabolism

2025/2/6 16:13:54

Objective: Myocardial inflammation during myocardial infarction (MI) could be inhibited by regulating arachidonic acid (AA) metabolism. Recent studies demonstrated that Sini Decoction (SND) was identified to be an effective prescription for treating heart failure (HF) caused by MI. But the anti-inflammatory mechanism of SND remained unclear. The work was designed to investigate the anti-inflammatory mechanism of SND through the AA metabolism pathway in vitro and in vivo experiments. Methods: An inflammatory injury model of H9c2 cells was established by lipopolysaccharide (LPS)-stimulated macrophage-conditioned medium (CM). The MI model was built by the ligation of left anterior descending (LAD) branch of coronary artery in rat. Meanwhile, the rats were divided into five groups: sham group, MI group, MI + Celecoxib group, MI + low-dose SND group (SND-L) and MI + high-dose SND group (SND-H). Cardiac function, histopathological changes and serum cytokines were examined four weeks later. Western blot analysis was conducted to verify the key enzymes levels in the AA metabolic pathway, including phospholipase A2 (PLA2), cyclooxygenases (COXs) and lipoxygenases (LOXs). Results: These in vivo results demonstrated that SND could improve the cardiac function and pathological changes of rats with MI, and regulate the key inflammatory molecules in the AA metabolism pathway, including sPLA2, COX-1, COX-2, 5-LOX and 15-LOX. In vitro, SND could decrease the release of proinflammatory cytokines including TNF-a and IL-6 and inhibit cell apoptosis in CM-induced H9c2 cells. Moreover, SND could protect H9c2 cells from the damage of CM by regulating nuclear factor kappa-B (NF-jB) signal pathway and the expression of COX-2. Conclusion: SND may be a drug candidate for anti-inflammatory treatment during MI by regulating the multiple targets in the AA metabolism pathway.

Intratumoral injection of two dosage forms of paclitaxel nanoparticles combined with photothermal therapy for breast cancer

2025/2/6 16:13:54

Objective: In order to enhance the efficacy of anti-breast cancer, paclitaxel nanoparticles (PTX NPs) and polypyrrole nanoparticles (PPy NPs) were combined with photothermal therapy and chemotherapy. At the same time, the two dosage forms of PTX NPs and PTX NPs gel were compared. Methods: PTX NPs were prepared by self-assembly method, and then the cytotoxicity in vitro was investigated by Methyl thiazolyl tetrazolium (MTT) and other methods, and the efficacy and side effects in vivo were further investigated. Results: The average hydrated diameter, PDI and electric potential of PTX NPs were (210.20 ± 1.57) nm, (0.081 ± 0.003) mV and (15.80 ± 0.35) mV, respectively. MTT results showed that the IC50 value of PTX NPs on 4 T1 cells was 0.490 lg/mL, while that of PTX injection was 1.737 lg/mL. The cell inhibitory effect of PTX NPs was about 3.5 times higher than that of PTX injection. The tumor inhibition rates of PTX NPs and gel were 48.64% and 56.79%, respectively. Together with local photothermal stimulation, the tumor inhibition rate of the PTX NPs reached 91.05%, surpassing that of the gel under the same conditions (48.98%), moreover, the organ index and H&E staining results of PTX NPs showed a decrease in toxicity. Conclusion: This combination therapy can significantly enhance the effect of anti-breast cancer, and the synergistic effect of chemotherapy and light and heat provides a feasible and effective strategy for the treatment of tumor.

Flavones in pomelo peel resist fibril formation of human islet amyloid polypeptide

2025/2/6 16:13:54

Objective: Exploring the formation and aggregation of human islet amyloid polypeptide (hIAPP) (amylin) fibers is significant for promoting the prevention and treatment of type II diabetes mellitus (T2DM). Flavones in pomelo peel have visible biological activity in the anti-diabetes aspect. The present study aimed to investigate the effects of five flavones [naringin (NRG), narirutin (NRR), nobiletin (NOB), sinensetin (SIN), and neohesperidin (NHP)] in pomelo peel on peptide aggregation and explore its possible mechanisms. The cell viability of flavones against peptide aggregation was also evaluated. Methods: The thioflavin T (ThT) assay and transmission electron microscopy (TEM) were used for evaluating the inhibition and disaggregation of flavones on peptide aggregation. The interaction mechanism was analyzed by endogenous fluorescence, molecular dynamics (MD) simulations, ultraviolet spectroscopy (UV) and isothermal titration calorimetry (ITC) experiments. The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and immune assays were performed to characterize the cell viability of flavones against peptide aggregation. Results: The five flavones showed a decrease in fluorescence intensity, fiber number and size under incubation with different molar ratios of hIAPP. The compounds can bind to the aromatic tyrosine (Tyr) residueTyr 37, resulting in the intrinsic fluorescence quenching of the peptides. Five flavones can form hydrogen bonds with hIAPP, which is likely to be based on their phenolic hydroxyl structure. They showed strong binding affinity with peptides. The reaction system of NRG and NRR observed an exothermic reaction, and the others were endothermic reactions. The absorption peaks of the compounds with hIAPP changed and showed hypochromic effects, indicating that there may be p-p stacking interaction. Flavones noticeably increased the cell viability in the presence of amyloid peptides and reduced the absorption intensity induced by peptide oligomers. Conclusion: A total of five flavones in pomelo peel have inhibitory and depolymerization effects on amyloid fibrils, and can significantly protect cells from the toxic effect of hIAPP and reduce the production of toxic oligomers.

Metabolome and transcriptome association study reveals biosynthesis of specialized benzylisoquinoline alkaloids in Phellodendron amurense

2025/2/6 16:13:54

Objective: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. Methods: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. Results: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. Conclusion: Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

Assembly and network of Rhei Radix et Rhizoma surface microbiome shaped by processing methods and sampling locations

2025/2/6 16:13:54

Objective: Rhei Radix et Rhizoma has five types of products, namely, raw rhubarb (RR), wine rhubarb (WR), vinegar rhubarb (VR), cooked rhubarb (CR), and rhubarb charcoal (RC). However, Rhei Radix et Rhizoma is easily contaminated with fungi and mycotoxins if not harvested or processed properly. Here, we intend to analyze how microbiome assemblies and co-occurrence patterns are influenced by sampling locations and processing methods. Methods: High-throughput sequencing and internal transcribed spacer 2 (ITS2) were carried out to study the diversities (a- and b-diversity), composition (dominant taxa and potential biomarkers), and network complexitity of surface fungi on RR, WR, VR, CR, and RC collected from Gansu and Sichuan provinces, China. Results: The phyla Ascomycota and Basidiomycota; the genera Kazachstania, Malassezia, and Asterotremella; and the species Kazachstania exigua, Asterotremella pseudolonga, and Malassezia restricta were the dominant fungi and exhibited differences in the two provinces and the five processed products. The a-diversity and network complexity were strongly dependent on processing methods. Chao 1, the Shannon index, and network complexity and connectivity were highest in the CR group. The adiversity and network complexity were influenced by sampling locations. Chao 1 and network complexity and connectivity were highest in the Gansu Province. Conclusion: The assembly and network of the surface microbiome on Rhei Radix et Rhizoma were shaped by processing methods and sampling locations. This paper offers a comprehensive understanding of microorganisms, which can provide early warning for potential mycotoxins and ensure the safety of drugs and consumers.

Piperine attenuates fibrogenesis and inflammation in hepatic fibrosis by switching STAT3 phosphorylation at Ser727

Xu Dai 1, Yunuo Wu 1, Lihua Lian,, Zhenyu Cui, Jixing Nan *, Yanling Wu ? — 2026/8/7 8:02:10

Objective: Piper nigrum is a medicinal and edible spice that is popular worldwide due to its unique spicy flavor, and is considered as a folk medicine for treating digestive system diseases. Piperine (PIP) is a bioactive alkaloid derived from P. nigrum, with remarkable hepatoprotective efficacy. Hepatic fibrosis represents a critical phase in the advancement of distinct chronic liver conditions towards cirrhosis, and currently lacks effective therapeutic drugs. The study aims to investigate the hepatoprotective functions of PIP and the underlying mechanism of PIP in combating hepatic fibrosis. Methods: C57BL/6J mice with hepatic fibrosis were induced by thioacetamide (TAA) and subsequently subjected to treatment with PIP or curcumin. Immortalized rat hepatic stellate cells (HSCs) were stimulated with transforming growth factor-b (TGF-b), followed by culture with PIP or niclosamide [signal transducer and activator of transcription 3 (STAT3) inhibitor], respectively. Human hepatic stellate cell line LX-2 were activated by TGF-b and transfected with specific small interfering RNA (siRNA) to silence STAT3 gene, and afterwards cultured with PIP. The mouse AML-12 cells and macrophage-like murine cells Raw 264.7 were stimulated with lipopolysaccharide (LPS), followed by culture with PIP. Results: In vivo, PIP reduced serum transaminase levels, collagen deposition, and decreased the excessive accumulation of extracellular matrix (ECM), comprising a-smooth muscle actin (a-SMA), collagen type I (Collagen I), and tissue inhibitor of metalloproteinases-1 (TIMP-1)/matrix metalloproteinase 13 (MMP13) ratio expressions. PIP inhibited inflammatory cytokines release and myeloperoxidase (MPO) expression, including interleukin-1 receptor type 1 (IL-1R1), cysteine-aspartic acid protease-1 (Caspase-1), and IL-6. PIP specifically inhibited STAT3 phosphorylation at Ser727, downregulated phosphorylated janus kinase 2 (p-JAK2), and upregulated suppressor of cytokine signaling 3 (SOCS3) and protein inhibitor of activated STAT 1/3 (PIAS1/3) expressions. In vitro, PIP inhibited ECM deposition and inflammatory cytokines release in activated HSCs. PIP selectively suppressed p-STAT3 at Ser727 without affecting Tyr705, and modulated p-JAK2, SOCS3, and PIAS1/3 expressions in activated HSCs, function as a STAT3 inhibitor. PIP inhibited LPS induced M1 polarization of RAW 264.7 macrophages. PIP decreased Cleaved Caspase 3 expression and apoptotic cells in LPS induced AML 12 hepatocytes. STAT3 deficiency amplified regulation of PIP on a-SMA and p-JAK2, as well as its upregulation on SOCS3 and PIAS1/3 in activated LX-2 cells. Conclusion: PIP improved hepatic fibrosis via inhibiting ECM excessive deposition and inflammatory secretion. Switching STAT3 phosphorylation at Ser727 might be the underlying targets for PIP against hepatic fibrosis, which provide an effective candidate and therapeutical strategy for hepatic fibrosis.Objective: Piper nigrum is a medicinal and edible spice that is popular worldwide due to its unique spicy flavor, and is considered as a folk medicine for treating digestive system diseases. Piperine (PIP) is a bioactive alkaloid derived from P. nigrum, with remarkable hepatoprotective efficacy. Hepatic fibrosis represents a critical phase in the advancement of distinct chronic liver conditions towards cirrhosis, and currently lacks effective therapeutic drugs. The study aims to investigate the hepatoprotective functions of PIP and the underlying mechanism of PIP in combating hepatic fibrosis. Methods: C57BL/6J mice with hepatic fibrosis were induced by thioacetamide (TAA) and subsequently subjected to treatment with PIP or curcumin. Immortalized rat hepatic stellate cells (HSCs) were stimulated with transforming growth factor-b (TGF-b), followed by culture with PIP or niclosamide [signal transducer and activator of transcription 3 (STAT3) inhibitor], respectively. Human hepatic stellate cell line LX-2 were activated by TGF-b and transfected with specific small interfering RNA (siRNA) to silence STAT3 gene, and afterwards cultured with PIP. The mouse AML-12 cells and macrophage-like murine cells Raw 264.7 were stimulated with lipopolysaccharide (LPS), followed by culture with PIP. Results: In vivo, PIP reduced serum transaminase levels, collagen deposition, and decreased the excessive accumulation of extracellular matrix (ECM), comprising a-smooth muscle actin (a-SMA), collagen type I (Collagen I), and tissue inhibitor of metalloproteinases-1 (TIMP-1)/matrix metalloproteinase 13 (MMP13) ratio expressions. PIP inhibited inflammatory cytokines release and myeloperoxidase (MPO) expression, including interleukin-1 receptor type 1 (IL-1R1), cysteine-aspartic acid protease-1 (Caspase-1), and IL-6. PIP specifically inhibited STAT3 phosphorylation at Ser727, downregulated phosphorylated janus kinase 2 (p-JAK2), and upregulated suppressor of cytokine signaling 3 (SOCS3) and protein inhibitor of activated STAT 1/3 (PIAS1/3) expressions. In vitro, PIP inhibited ECM deposition and inflammatory cytokines release in activated HSCs. PIP selectively suppressed p-STAT3 at Ser727 without affecting Tyr705, and modulated p-JAK2, SOCS3, and PIAS1/3 expressions in activated HSCs, function as a STAT3 inhibitor. PIP inhibited LPS induced M1 polarization of RAW 264.7 macrophages. PIP decreased Cleaved Caspase 3 expression and apoptotic cells in LPS induced AML 12 hepatocytes. STAT3 deficiency amplified regulation of PIP on a-SMA and p-JAK2, as well as its upregulation on SOCS3 and PIAS1/3 in activated LX-2 cells. Conclusion: PIP improved hepatic fibrosis via inhibiting ECM excessive deposition and inflammatory secretion. Switching STAT3 phosphorylation at Ser727 might be the underlying targets for PIP against hepatic fibrosis, which provide an effective candidate and therapeutical strategy for hepatic fibrosis.

6-Gingerol alleviates hepatic fibrosis via inhibiting glycolytic reprogramming by targeting IGF2BP2

2026/8/7 8:02:10

Objective: Hepatic fibrosis remains a major global health burden, often progressing to cirrhosis and hepatocellular carcinoma. Despite extensive research, no effective antifibrotic drugs are clinically approved due to the complexity of hepatic stellate cells (HSCs) activation and metabolic remodeling. Enhanced glycolysis is increasingly recognized as a metabolic hallmark sustaining the profibrotic phenotype of HSCs, representing a key but underexplored therapeutic target. This study explored the antifibrotic effect of 6- gingerol, the principal bioactive compound of Zingiber officinale, and revealed its novel mechanism involving metabolic-epitranscriptomic regulation of HSC activation. Methods: Liver fibrosis was induced in C57BL/6 mice by carbon tetrachloride (CCl4) and bile duct ligation (BDL), followed by oral 6-gingerol treatment. Complementary in vitro studies were performed in transforming growth factor-b1 (TGF-b1)-stimulated Lieming Xu-2 (LX-2 cells, a human hepatic stellate cell line) and primary HSCs from liver fibrosis mice. Integrated transcriptomics, network pharmacology, molecular docking, and cellular thermal shift assays (CETSA) were combined with functional genetic manipulation to identify and verify molecular targets. Results: 6-Gingerol alleviated hepatic injury and collagen deposition in both CCl4- and BDL-induced models and improved liver function. Mechanistically, 6-gingerol inhibited glycolysis by downregulating hexokinase 2 (HK2), pyruvate kinase M2 (PKM2), and lactate dehydrogenase A (LDHA), normalizing pyruvate and lactate metabolism, and restoring mitochondrial respiration. Insulin-like growth factor 2 mRNAbinding protein 2 (IGF2BP2) was identified as an important mediator associated with glycolytic enzyme expression and HSCs activation, and its silencing abolished 6-gingerol’s antifibrotic effects. Conclusion: Our study demonstrates that 6-gingerol attenuates hepatic fibrosis by targeting IGF2BP2- dependent glycolytic reprogramming, thereby restoring metabolic homeostasis and inhibiting HSCs activation. These findings uncover a novel metabolic-epitranscriptomic mechanism and suggest that 6- gingerol represents a promising therapeutic strategy for chronic liver diseases.

Formononetin alleviates ovarian fibrosis and inflammation via targeting SOCS3 and inhibiting TGF-b1/Smad signaling in PCOS

2026/8/7 8:02:10

Objective: Polycystic ovary syndrome (PCOS) is an endocrine and metabolic disorder characterized by ovulatory dysfunction. Formononetin (FMN), a bioactive compound derived from leguminous plants, including Glycyrrhiza uralensis, Trifolium pratense, and Astragalus membranaceus, exhibits antiinflammatory and anti-fibrotic properties. This study aimed to evaluate the therapeutic effects of FMN in a PCOS rat model and elucidate its underlying molecular mechanisms. Methods: A PCOS-like model was established in 3-week-old female Sprague-Dawley rats by daily dehydroepiandrosterone (DHEA) injections. The efficacy of FMN was assessed by monitoring estrous cycles, hormone levels, glucose tolerance, and ovarian tissue morphology. Network pharmacology and transcriptome sequencing were used to predict the molecular targets and signaling pathways of FMN. The binding and stabilizing effects of FMN on suppressor of cytokine signaling 3 (SOCS3) were confirmed using molecular docking, molecular dynamics (MD) simulations, and cellular thermal shift assay (CETSA)-Western blotting (WB) (CETSA-WB). The expression of SOCS3, inflammation- and fibrosis-related markers, and the components of the transforming growth factor beta1 (TGF-b1)/Smad pathway were quantified by quantitative polymerase chain reaction (qPCR), WB, and immunohistochemistry. Functional validation was performed using SOCS3 small interfering RNA (siRNA). Results: DHEA successfully induced a PCOS-like phenotype, including disrupted estrous cycle, hyperandrogenism, an elevated luteinizing hormone to follicular stimulating hormone (LH/FSH) ratio, insulin resistance, and abnormal ovarian morphology. FMN treatment significantly reversed these abnormalities and reduced ovarian collagen deposition. Network pharmacology and transcriptome analyses indicated that the therapeutic efficacy of FMN was primarily attributable to its anti-fibrotic and antiinflammatory properties, with SOCS3 identified as a potential key molecule. Molecular docking and MD simulations, along with CETSA-WB, further revealed that FMN directly binds to SOCS3, enhancing its stability and thereby inhibiting ovarian fibrosis in PCOS. Moreover, qPCR, WB, and immunohistochemistry confirmed that FMN regulates the TGF-b1/Smad signaling pathway. Importantly, SOCS3siRNA abolished fibrotic-related collagen levels and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) expression, supporting the notion that FMN ameliorates PCOS-induced ovarian fibrosis via SOCS3. Conclusion: FMN alleviated ovarian dysfunction and hyperandrogenism in DHEA-induced PCOS rats by stabilizing SOCS3 and inhibiting TGF-b1/Smad signaling, thereby attenuating ovarian fibrosis and inflammation. This reveals a novel SOCS3-centered mechanism for FMN and underscores its therapeutic potential for PCOS.

Huashi Baidu formula may attenuate adriamycin-induced myocardial injury via modulating NRF2-ATF3-SRXN1 axis-mediated ferroptosis

2026/8/7 8:02:10

Objective: Myocardial injury is a fatal adverse effect of adriamycin (ADR), which greatly limits its clinicalapplication. Numerous studies have shown that Huashi Baidu formula (HBF) may exert a satisfactoryeffect on myocardial function repair. This study aims to reveal the underlying mechanisms and mainbioactive compounds (BACs) of HBF against myocardial injury. Methods: Firstly, this study established an ADR-induced mouse model to assess the protective effects ofHBF against myocardial injury. Subsequently, transcriptomic profiling was conducted to screen the differentially expressed genes among the HBF treatment, ADR-induced model, and normal control groups.Following the construction and analysis of protein–protein interactions, gene set enrichment analysis,and co-expression matrix, the key targets of HBF against myocardial injury were screened and further validated based on the ADR-induced mouse model. An integrative approach combining network mapping,molecular docking, and molecular dynamics simulation was performed to identify the representative BACs. Results: Based on an ADR-induced myocardial injury mouse model, HBF significantly ameliorated myocardial injury by improving body weight loss, decreasing mortality, and rescuing severe cardiac fibrosis. A totalof 361 ADR-related genes and 42 HBF therapeutic effect-related genes were identified with the thresholdsP < 0.05 and fold change (FC) > 1.2 or < 0.833. Transcriptomic profiling-based networks demonstrated thatthe nuclear factor erythroid 2-related factor 2 (NRF2)-cyclic AMP-dependent transcription factor ATF-3(ATF3)-sulfiredoxin-1 (SRXN1) axis-mediated ferroptosis was the key target of HBF against myocardialinjury. In vivo validation showed that HBF treatment effectively inhibited serum enzymatic biomarkers ofgeneral myocardial injury (aspartate aminotransferase, lactate dehydrogenase, and creatine kinase), myofi- broblast activity (transforming growth factor beta 1 and a-smooth muscle actin), inflammatory cytokines(tumor necrosis factor-a and interleukin-6), oxidative damage (superoxide dismutase, malondialdehyde,accumulated iron content, and TUNEL-positive cells). Mechanically, HBF may restore the dysregulation ofthe NRF2-ATF3-SRXN1 signal axis, leading to inhibition of ferroptosis-associated protein expression.Emodin, rhein, and lactiflorin were identified as the underlying representative BACs of HBF against myocardial injury due to the strong binding affinities between candidate BACs and key targets [Kelch-like ECHassociated protein 1 (KEAP1), ATF3, SRXN1, system Xc- (SLC3A2/SLC7A11), glutathione peroxidase 4(GPX4), ferroportin (FPN), glutathione synthetase (GSS)]. Molecular dynamics simulations further verifiedthese findings, revealing the stable binding among the three components and the key targets. Conclusion: HBF may attenuate ADR-induced myocardial injury via modulating NRF2-ATF3-SRXN1 axismediated ferroptosis, and emodin, rhein, and lactiflorin are potential representative BACs.

A cross-cultural ethnobotanical assembly highlights diverse health-promoting functions of genus Panax globally

2026/8/7 0:00:00

Objective: This study aims to achieve a cross-cultural consolidation of the local and traditional knowledge in order to contribute to an understanding of these knowledge systems and to guide bioprospecting on Panax spp. Methods: Documentation-based taxonomic revision was performed to define the current accepted spe-cies in Panax. Historic uses of Panax spp. used in China was compiled using historical herbal literature. A cross-cultural ethnobotanical assembly was performed to compile the traditional uses of Panax spp. globally. With a network-based integration, the scattered local and traditional knowledge was consoli-dated and evaluated. Results: Currently, the genus Panax comprises 14 accepted species, mainly distributed in East Asia and eastern North America, all of which are used locally as traditional food and/or medicine. Four species are recorded in Chinese historical herbals. Eight species and one variety are used by 25 of the Chinese ethnic minorities. Beyond China, there are ten species and two varieties used traditionally in 12 countries. General health promotion functions are frequently mentioned. Conclusion: In different biocultural backgrounds, the used species and usages varied greatly, and even the same species can be used differently. For the first time, this study archives a comprehensive understanding on the health benefits of global Panax spp. and provides solid evidence for their versatility in main-taining human health from the perspective of ethnobotany.

Integrated network pharmacology, transcriptomics and experimental validation to explore mechanisms of Wenyang Jiedu Granule on IAVinduced pneumonia

2026/5/18 16:30:25

Objective: Wenyang Jiedu Granule (WYJD) is an effective traditional Chinese medicine (TCM) preparation that has been generally applied for treating respiratory infectious diseases. Clinical observations involving thousands of cases have demonstrated that WYJD could alleviate disease progression and improve symptoms in treating respiratory viral infections, including SARS-CoV-2 and influenza virus. However, the chemical basis and underlying mechanisms of WYJD against influenza A virus (IAV)-induced pneumonia remain to be elucidated. This study aimed to reveal the underlying mechanisms of WYJD in treating IAV-induced pneumonia by a combined strategy of network pharmacology, transcriptomics and experimental validation. Methods: The pneumonia model was established in BALB/c mice via infection with H1N1 IAV to evaluate the therapeutic effects of WYJD on IAV-induced pneumonia. Firstly, ultra-high performance liquid chromatography-quadrupole Exactive Orbitrap mass spectrometer/tandem mass spectrometer (UPLC-Q Exactive Orbitrap-MS/MS) was employed to analyze the main chemical components in WYJDcontaining serum. Subsequently, the effects of WYJD on IAV-induced pneumonia were assessed through pathological observation, plaque forming assay, biochemical analysis, Evans blue staining assay, and immunofluorescence assay. Mechanistically, an integrated approach of network pharmacology and transcriptomics was applied to explore the potential active components, targets and related pathways of WYJD against IAV-induced pneumonia. Fluorescence TUNEL assay, quantitative real-time PCR (qRTPCR) and Western blotting were utilized for experimental validation and mechanistic studies. Results: Using UPLC-Q Exactive Orbitrap-MS/MS, a total of 25 prototypes and 15 metabolites were identified in the serum of mice after WYJD administration. WYJD treatment showed protective effects on IAVinduced pneumonia by inhibiting inflammation and lung barrier damage in the IAV-induced pneumonia mice model. Network pharmacology combined with transcriptomics analysis indicated that WYJD exerted therapeutic effects against IAV-induced pneumonia mainly through the synergistic effects of 11 active components, which regulated ten critical signaling pathways via 86 targets. Further experimental validation demonstrated that WYJD could alleviate IAV-induced pneumonia via the IL-17 signaling pathway, Toll-like receptor 7 (TLR7)/Myeloid differentiation primary response dene 88 (MyD88)/mitogen-activated protein kinases (MAPKs)/activator protein 1 (AP-1) signaling pathway and apoptosis. Conclusion: This study revealed the main active components and mechanisms of WYJD against IAVinduced pneumonia through the IL-17 signaling pathway, TLR7/MyD88/MAPKs/AP-1 signaling pathway and apoptosis, which provides novel insights into the clinical application of WYJD in treating influenza and its complications.

Combined network pharmacology and biological verification to investigate anti-influenza A virus effect and potential mechanism of Pudilan Xiaoyan Oral Liquid

2026/5/18 16:30:25

Objective: Influenza is a common respiratory disease affecting human health and life. Pudilan Xiaoyan Oral Liquid (PDL) has frequently been used to treat respiratory infections such as COVID-19, pharyngitis, and tonsillitis. However, its efficacy against influenza A virus (IAV) infection remains unclear. This study investigates the anti-IAV activity and potential mechanism of action of PDL. Methods: The in vitro anti-IAV activity was investigated in a virus-infected MDCK (Madin-Darby canine kidney) cell model. The in vivo protective effect on AIV infection was evaluated in a virus-infected mice model. qRT-PCR was performed to examine the anti-inflammatory effects of the PDL. A network pharmacology method was used to investigate the underlying mechanism of PDL against IAV. Western blotting was performed to confirm the antiviral mechanism of PDL against IAV. Results: Our findings indicated that PDL had a broad-spectrum anti-IAV effect in vitro and exhibited an excellent protective effect against IAV infection by significantly increasing the survival rate, extending the survival time, and reducing pathological lung tissue damage. Moreover, PDL effectively inhibited the mRNA expression of inflammatory factors in lung tissue. GO analysis revealed that the potential targets were primarily associated with defense response regulation, positive regulation of cytokine production, and positive regulation of responses to external stimulation. PPI analysis indicated that Sarcoma (SRC), signal transducer and activator of transcription 3 (STAT3), mitogen-activated protein kinase1 (MAPK1), phosphatidylinositol 3-kinase regulatory subunit alpha (PIK3R1), and epidermal growth factor receptor (EGFR) might be potential PDL targets against IAV infection. Mechanistically, inhibition of the Toll-like receptor 3 (TLR3)/myeloid differentiation primary response gene (88) (MyD88) signaling pathway to attenuate an excessive inflammatory response might be one of the primary mechanisms of PDL against IAV. Conclusion: PDL efficiently suppressed in vitro IAV replication and provided protection against lethal influenza virus infection in vivo. Network pharmacology analysis revealed that the key PDL targets against influenza might be highly associated with biological processes, such as regulation of defense responses, cytokine production, and the Toll-like receptor signaling pathway. Mechanistically, regulating the TLR3/MyD88 signaling pathway may be one of the primary mechanisms of PDL against influenza virus infection. These findings indicate that PDL could be a potential therapeutic option for the clinical treatment of influenza viral infections.

Retrospective cohort study on Shufeng Jiedu granules for SARS-CoV-2 infection: Real-world evidence from multicenter outpatient clinics in a predominantly caucasian population

Thomas Friedemann a,1, Uta Knierim a,b,1 ,, Zhang Wei c,d,, Sven Schr?der a,b — 2026/5/18 16:30:25

Objective: Due to the lack of clinical evidence on the effectiveness and safety of Shufeng Jiedu (SFJD) Granules in Europe, it is unclear if SFJD can effectively and safely be used as an adjunct treatment for mild SARS-CoV-2 infection in a predominantly Caucasian outpatient population in Germany. Therefore, we conducted a retrospective cohort study to evaluate the real-world effectiveness and safety of SFJD compared to standard symptomatic care. Methods: This retrospective cohort study included adult patients (≥ 18 years) with mild SARS-CoV-2 infection symptoms treated between October 2021 and March 2023 across four outpatient clinics in Hamburg, Germany. Patients were allocated to either the SFJD group or a control group receiving standard symptomatic care. Clinical data, including symptom duration, intensity [measured by Visual Analogue Scale (VAS)], and adverse events, were analyzed. The primary endpoint was the time to total symptom alleviation (TTSA), defined as the first 24–hour period with all index symptoms rated VAS ≤ 1 and no temperature ≥ 37.0 °C. Secondary outcomes included symptom-specific duration and intensity. Data were verified by propensity score matching (PSM). Results: A total of 81 patients met inclusion criteria, with 38 patients assigned to the SFJD group and 43 patients assigned to the control group. Baseline characteristics were comparable between groups. The SFJD group showed a statistically significant different Time to Total Symptom Alleviation (TTSA) compared to controls [(5.76 ± 3.74) d and (11.50 ± 7.45) d, difference of 5.74 d, P < 0.001]. Symptomspecific improvement was most pronounced for rhinorrhea, fatigue, fever, and shortness of breath. Intensity scores were also lower in the SFJD group for multiple symptoms. No severe adverse events were observed. Gastrointestinal symptoms were equally distributed and not clearly attributable to treatment. 1:1 Nearest-neighbor PSM confirmed the TTSA results. Conclusion: SFJD Granules demonstrated beneficial effects in reducing the duration and intensity of mild SARS-CoV-2 infection symptoms in a real-world outpatient setting, without significant safety concerns. These findings support the potential of SFJD as an adjunctive therapy for early-stage COVID-19, especially in populations outside of East Asia. Prospective randomized trials are warranted to confirm these results.

Exploring possible mechanisms of antiviral effect of Shufeng Jiedu Capsule on viral pneumonia: A comprehensive analysis

2026/5/18 16:30:26

Objective: Influenza A virus (IAV) is a major cause of respiratory tract infections. Shufeng Jiedu Capsule (SFJD) has a broad-spectrum antiviral effect, however, it is not clear whether SFJD acts by targeting the immune-inflammatory pathway. The aim of this study was to elucidate the therapeutic effect and the mechanism of antiviral action of SFJD in mice with influenza virus pneumonia. Methods: In this study, a mouse model of influenza virus pneumonia was used to evaluate the efficacy of SFJD in the treatment of influenza virus pneumonia. Next, the potential pathways of action by which SFJD exerts its antiviral effects were analysed in conjunction with network pharmacology and and untargeted metabolomics. Finally, in vivo experimental validation was conducted through protein immunoblotting, transmission electron microscopy, and immunofluorescence experiments. Results: SFJD significantly reduced lung index, lowered viral load, attenuated lung tissue lesions, modulated inflammatory metabolism in lung tissue, reduced macrophage inflammatory infiltration and protected lung epithelial cells in mice. The results of the network pharmacological analysis and metabolomics analysis suggested that SFJD might act through the Toll-like receptor 4(TLR4)/myeloid differentiation primary response protein 88(MyD88)/ TNF receptor associated factor (TRAF6)/nuclear factor kappa-B(NF-jB) signalling pathway. SFJD was able to down-regulate the expression of TLR4, MyD88, TRAF6, and NF-jB proteins in lung tissues. Conclusion: SFJD could effectively attenuate lung injury induced by influenza virus infection and provide a scientific basis for the protection of respiratory mucosal epithelial cells. SFJD could effectively alleviate lung injury by regulating the TLR4/MyD88/TRAF6/NF-jB signaling pathway, inhibiting the levels of TNFa, IL-6, which may be the key mechanism for its antiviral effects.

Yu-Ping-Feng-San mitigates development of emphysema and its exacerbation induced by influenza virus in mice

2026/5/18 16:30:26

Objective: Yu-Ping-Feng-San (YPFS) has been found to significantly reduce exacerbation rate and the risk of a second exacerbation in patients with chronic obstructive pulmonary disease (COPD). However, the effect of YPFS on emphysema development and its exacerbation caused by influenza virus is still unclear. This study aims to investigate the protective effect of YPFS on the impaired lung function and increased inflammation that occurs during the development and exacerbation of emphysema. Methods: This study developed an emphysema mice model using porcine pancreatic elastase (PPE) and treated it with YPFS [1 950, 975, and 487.5 mg/(kg d)] once daily. On day 7 post-PPE challenge, peripheral leucocytes and inflammatory cells in lungs were analyzed, respectively. Pulmonary inflammatory mediators were determined. On day 28, invasive lung function and inflammatory mediators were measured. In addition, histopathological changes at both time points were assessed. We also developed an exacerbation of emphysema mouse model by intratracheally infected mice with influenza H1N1 virus on day 28. After infection, YPFS administration was discontinued, and the protective effect was determined by lung function, viral titer and cytokine levels in the lungs, and lung histopathological changes on day 5 postinfection. Results: The results demonstrated that 32 days of YPFS administration significantly improved lung function and reduced severity of emphysema in PPE-treated mice. It significantly reduced monocytes and neutrophils, while increased lymphocytes in peripheral blood. Additionally, YPFS inhibited the accumulation of alveolar macrophages and the expression of cytokines in mice treated with PPE. MMP-9, fibronectin, and VEGF, which are inflammatory mediators associated with airway remodeling, were also inhibited by YPFS. The results also demonstrated that YPFS prophylaxis prior to viral infection benefited mice treated with PPE and H1N1 as evidenced by decreased lung cytokine levels, lowered lung index, and improved lung histopathology. Conclusion: YPFS prevents the development of emphysema and its exacerbation induced by influenza virus in mice. Our finding provides scientific evidence for the prophylaxis application of YPFS in mitigating acute exacerbation of chronic obstructive pulmonary disease (AECOPD).

Efficacy and safety of Toujie Quwen Granules in patients infected with SARS-CoV-2: A prospective cohort study of 156 cases

2026/5/18 16:30:26

Objective: Given traditional Chinese medicine (TCM)’s efficacy in influenza and severe acute respiratory syndrome (SARS), it was adopted clinically in the early stage of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection outbreak in China. However, the therapeutic effects of Toujie Quwen (TJQW) Granules in treatment of (SARS-CoV-2) infection have not been fully elucidated. This study aimed to investigate the efficacy and safety of TJQW in patients infected with SARS CoV-2 and provide evidence for clinical practice of TCM. Methods: A cohort study was conducted on SARS-CoV-2 infection admitted between 20 January and 15 March 2020. Patients were divided into two cohorts based on their clinical medication plan: TJQW combined with Western medicine (TJQW + WM) group and Western medicine (WM) group. The WM group received conventional WM treatment (antibiotics, glucocorticoids, oxygen therapy, arbidol, chloroquine, lopinavir/ ritonavir, ambroxol hydrochloride), while the TJQW + WM group received TJQW in addition to WM (15 g/bag, two bags at a time, three times a day). The incidence of severe disease and chest computed tomography (CT) findings before and after treatment were compared between groups. Dynamic analysis was performed on white blood cell (WBC) count, lymphocyte count (LYM), lymphocyte percentage (LYM%), Krebs von den Lungen 6 (KL-6) and symptom scores within 15 d of enrolment. Results: Six cases in the TJQW + WM group progressed to severe disease, with an incidence of 7.89% (6/76), while 16 cases in the WM group progressed to severe disease, with an incidence of 20% (16/80, P = 0.025), and the difference was significant. The CT scores of the TJQW + WM group decreased continuously from day 6 of disease onset and were significantly lower than those of the WM group on days 12 and 15, with significant differences. On day 12 of the disease, the improvements in WBC and LYM in the TJQW + WM group were superior to those in the WM group, with significant differences. The KL-6 levels of the two groups were compared by fitting the data to a generalised linear model. After controlling for the effects of time from onset to detection, the KL-6 level of the TJQW + WM group was significantly lower than that of the WM group (t = 3.703, P < 0.01). Repeated measures analysis of variance was performed on the total score of major symptoms and showed insignificant differences (F = 0.031, P = 0.871). However, there was an interaction effect between treatment plan and treatment duration, with significant differences in the degree of change at each time point for the global scores (F = 6.502, P < 0.01). The TJQW + WM group showed a better improvement in symptoms and a reduction in the expression of inflammatory factors than the WM group. Conclusion: TJQW could effectively decrease the incidence of severe disease, reduce pulmonary inflammation and fibrosis, and alleviate clinical symptoms.

Research progress of polysaccharide detection methods in traditional Chinese medicine

2026/5/18 16:30:26

In recent years, with the advancement of analytical technology and the development of molecular biology, the research of traditional Chinese medicine (TCM) polysaccharides is becoming a new frontier and hot spot in life science research due to their effectiveness and non-toxicity. The quantitative determination of polysaccharides plays an extremely important role in the quality control of polysaccharide drugs. However, the structural complexities of natural polysaccharides present a major obstacle to their accurate quantification. The objective of this review is to provide a comprehensive guide for the accurate quantification of polysaccharides in TCM. In this paper, the quantitative methods of polysaccharides are divided into traditional quantitative methods and new methods, and summarized from the medicinal materials included in the Chinese Pharmacopoeia (2025 edition) and the relevant literature in the past ten years. The traditional methods for polysaccharide quantification include titration, colorimetry, high performance liquid chromatography (HPLC) and resonance light scattering (RLS). With the technological progress, new nondestructive testing methods such as near-infrared spectroscopy (NIR) and hyperspectral imaging (HSI) have gradually emerged. In addition, glycospectrometry, as another new method, can accurately quantify the structural characteristics and content of polysaccharides by combining positional enzyme digestion technology and HPLC. Moreover, this review elaborates on the principles and discusses the advantages and limitations of each method mentioned. This paper aims to provide a comprehensive reference for the accurate determination of polysaccharide content in TCM, enhance its practicability and precision, and promote its wider application in the quality control of TCM and Chinese patent medicine preparations.

Isoquinoline alkaloids in Coptis chinensis to treat Alzheimer’s disease through promoting growth of Bifidobacterium breve inhibiting abnormal autophagy using a novel AI high-content intelligent imaging system

2026/5/18 16:30:27

Objective: Coptis chinensis has been shown to increase beneficial intestinal bacteria and treat Alzheimer’s disease (AD). Bifidobacterium breve can effectively treat AD through the gut-brain axis. Therefore, this study aimed to study the joint effects of C. chinensis and B. breve in the treatment of AD. Methods: 16S rRNA was used to test the abundance of bacterial flora in APPswe/PS1DE9 mice after C. chinensis administration. To determine the efficacy of C. chinensis combined with B. breve on AD, pathological section staining, Barnes maze, Western blotting and ELISA were utilized to confirm the improvement of cognitive dysfunction in AD mice after administration. In order to elucidate the pharmacodynamic components of monomers in C. chinensis, network pharmacology was used to screen the components related to autophagy and confirm the pharmacodynamic effects by artificial intelligence (AI) high-content intelligent imaging. Results: The results of 16S rRNA sequencing indicated that C. chinensis could modulate the abundance of B. breve in AD mice. Pathological assessments, Barnes maze testing, and additional experiments have shown that C. chinensis combined with B. breve can improve the memory and learning ability of AD mice by reducing neuronal apoptosis and amyloid-b (Ab) peptide deposition. Network pharmacology combined with AI high-content intelligent imaging technology and Western blotting experiments demonstrated that magnoflorine, 13-methylberberine and palmatrubine in C. chinensis could exert neuroprotective effects in AD mice by up-regulating p62 protein expression while down-regulating Beclin-1 and microtubule-associated protein 1 light chain 3 II (LC3II) protein levels, thereby modulating autophagy-related pathways. Conclusion: C. chinensis can enhance the abundance of beneficial bacteria in the gut and ameliorate cognitive dysfunction in AD mice by interacting with B. breve. Moreover, magnoflorine, 13-methylberberine, and palmatrubine within C. chinensis can also mitigate excessive autophagy and oxidative stress in nerve cells.

Integrated network pharmacology and metabolomics to analyze the mechanism of Shaofu Zhuyu Decoction in treatment of dysmenorrhea of cold coagulation and blood stasis

Yilu Luo a,1, Zengguang Wu b,1, Ruifeng Ji a, Lingling Shen c, Xin He a,d,e — 2026/5/18 16:30:27

Objective: To study the mechanism of Shaofu Zhuyu Decoction (SFZYD) in treating primary dysmenorrhea (PD) with cold coagulation and blood stasis based on metabonomics and network pharmacology. Methods: The disease model of primary dysmenorrhea in rats with cold coagulation and blood stasis was established according to the previous research methods. The model of cold coagulation and blood stasis was built with ice water bath, and the model of PD was built with estradiol and oxytocin injection.The success of modeling was judged according to the physiological condition of rats, the change of body weight, the number of twisting and the change of uterine blood flow. UPLC-Q/TOF MS technique was used to identify the components and their main components in plasma in the Wenjing group (WJ, means warming channel), Huoxue group (HX, means activating blood), and Quanfang group (QF, means complete prescription) group, and a ‘‘compound-target-pathway” network was constructed by network pharmacology. Untargeted metabolomics studies were applied to screen out endogenous differential metabolites and analyze their related metabolic pathways. The analgesic effect, pathological morphology and inflammatory mediators were evaluated, and the pathway targets were verified. Results: A total of 93 chemical components and 22 components in plasma were identified. A total of 52 potential targets of SFZYD for primary dysmenorrhea were identified by network pharmacological analysis, and 12 endogenous differential metabolites and four related metabolic pathways were screened out by metabonomics. The possible targets and pathways of action of the components of SFZYD into blood were analyzed by network pharmacology, and the results showed that it can widely act on the proteins and pathways related to thrombosis, new angiogenesis, vasodilation, inflammatory immune response and central sedation. Body torsion test showed that drug administration could reduce the frequency of body twisting in dysmorrhea rats, ELISAs showed that SFZYD could inhibit the release of inflammatory factors TNF-a and IL-6. Western blotting showed that SFZYD could reduced the protein expression of COX-2, p-ERK2 in uterine tissue, inhibiting inflammatory cell infiltration and inflammatory factor expression. By down-regulating the expression of p-MLC protein in uterine tissue, the strong contraction of smooth muscle was inhibited, thus relieving dysmenorrhea. Conclusion: Based on the research idea of ‘‘disease-syndrome-prescription-medicine”combination, this study discussed the internal relationship of SFZYD’s drug property, functional taste and efficacy. The substance basis of ‘‘warm property and pungent taste” in the WJ group reflects pungent taste is dispersive and propulsive, the warm property can dispel cold, to help Yang promoting circulation of qi and dispersing accumulation of pathogen, dredging connection tissue, dispersing fluid and promoting circulation of blood. Thus, it has the effect of WJ dispelling cold, regulating the circulation of qi and alleviating pain. The substance basis of ‘‘warm property, pungent and bitter taste” in HX group reflects that pungent taste is dispersive and propulsive, bitter taste can resolve stasis, warm property can dispel cold. All result in some effects of blood circulation, promoting qi, repelling congestion and improving dysmenorrhea. This study revealed the multi-component, multi-target and multi-pathway integration mechanism of SFZYD in the treatment of PD with cold coagulation and blood stasis, which preliminarily clarify scientific connotations about property theory of pungent, bitter taste and warm property,efficacy and material basis, laid a foundation for further research on the mechanism of action of SFZYD in the treatment of PD.

A methodological approach for complex mixture aqueous solution analysis by ATR-FTIR spectroscopy

Lingyu Han a,1, Qun Zhou b,1, Hang Yin a,b, Suqin Sun b — 2026/5/18 16:30:27

Objective: To develop an in situ, nondestructive approach for simultaneously acquiring overall structural and intermolecular interaction information of multiple components in complex aqueous mixtures without sample separation. Methods: In this paper, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) was utilized to detect and analyze the juice and decoction (clinical application forms) of different processed products of a traditional Chinese medicine (TCM)—Rehmanniae Radix (Dihuang in Chinese). By the analysis of IR and the second derivative IR (SD-IR) spectra, the spectral structure information of water, organic acid, stachyose, glucose, galactose, fructose, manninotriose, amino acid and protein in the mixture system was extracted through non separation fingerprint characteristic. Results: A large number of in situ dynamic tracked IR spectra, obtained by ATR-FTIR and TimeBase software, were used to excavate the changes of absorption peak intensity (structural analysis) and diffusion velocity (time resolution) in the process of liquid adsorption-diffusion. The results indicated the coexistence of free hydroxyl groups, intermolecular hydrogen bonds, and intramolecular hydrogen bonds, analyzed by two-dimensional correlation infrared spectroscopy (2DCOS-IR), confirming their complex and dynamic hydrogen-bonding network structures. Furthermore, spectral analysis revealed interactions among the various active components in the mixtures, providing a microstructural basis for interpreting their holistic effects. Conclusion: This study provided a theoretical basis for the efficacy and pharmacologic mechanism of different processed forms of TCM from the perspective of physical chemistry. Also, it discussed how to carry out nondestructive and in situ dynamic tracking of complex mixture aqueous solution system, and how to study the timeliness, synergy and interaction of samples.

Optimization of imperatorin extraction from Qianliexin Capsules using deep eutectic solvent

2026/5/18 16:30:27

Yuqing Meng a

Shuxuening injection for acute ischemic stroke: Efficacy and safety − a randomized, double-blind, placebo controlled, multi-center clinical trial

2026/5/18 16:30:27

Objective: To evaluate the efficacy and safety of Shuxuening injection, a Chinese remedy from Ginkgo biloba, in treating patients with acute ischemic stroke (AIS). Methods: A randomized, double-blind, placebo-controlled, multi-center clinical trial was conducted from 21 November 2018 to 20 January 2022 in 31 participating research centers involving 446 patients with AIS. Patients received either a Shuxuening injection (experimental group) or a placebo (control group) alongside basic treatment (aspirin enteric-coated tablets). The primary efficacy endpoint was the percentage of patients with a modified Rankin scale (mRS) score ≤ 2 at 90-day post-stroke. The secondary efficacy endpoints were the changes in mRS, National Institute of Health stroke scale (NIHSS), and Barthel index (BI) scores, and the incidence of adverse events. Results: The percentage of patients with mRS scores ≤ 2 at 90-day post-stroke was significantly higher in the experimental than in the control (P < 0.05) group. The percentage of patients with scores ≤ 1 at 90- day post-stroke was higher in the experimental group (P < 0.05). The change value of NIHSS scores in the experimental group was higher at 10-day post-treatment and 90-day post-stroke (P < 0.05). The percentage of NIHSS score reduction ≥ 50% was higher in the experimental group at 90-day post-stroke (P < 0.05). The change value of BI score and percentage of BI score ≥ 75 were higher in the experimental group at 90-day post-stroke (P < 0.05). Safety profiles did not differ between the groups. Conclusion: Shuxuening injection effectively improves the functional outcome of patients, reduces disability rates, improves neurological deficits, enhances activities of daily living, and demonstrates good safety profiles.

Novel step-by-step strategy for evaluating treatment of traditional Chinese medicine injections for community-acquired pneumonia in adults

2026/5/18 16:30:28

Objective: Community-acquired pneumonia (CAP) is a leading cause of death from infectious diseases in adults worldwide. Traditional Chinese medicine injections (TCMIs) are often used as complementary therapies. This study aimed to evaluate the efficacy and safety of TCMIs for the treatment of CAP. Methods: A step-by-step strategy was first designed including data collection, systematic review, Bayesian network meta-analysis (NMA), and assessment of the NMA. Eight databases were searched for relevant randomized clinical trials (RCTs) from their inception to December 31, 2022. The Cochrane risk-of-bias tool was used to assess the methodological quality and the risk of bias of the included RCTs. Bayesian NMA was performed by WinBUGS 14.0 and STATA 15.0. Finally, the Grading of Recommendation, Assessment, Development, and Evaluation (GRADE) system was used to evaluate the quality of evidence for each outcome. Results: A total of 35 RCTs involving 4 291 participants were included. For clinical effectiveness rate, the Reduning Injection (RDN) combined with Western medicine (WM) was the most likely to be the best treatment (SUCRA = 0.919). And the top three TCMIs interventions were RDN + WM, Xiyanping Injection (XYP) + WM, Tanreqing injection (TRQ) + WM. For defervescence time, disappearance time of cough, and the disappearance time of lung rales, the XYP with WM was probably the best treatment (SUCRA = 0.822, 0.838, 0.816). Moreover, adverse drug reactions have been reported in 14 RCTs. Conclusion: The TCMIs can effectively relieve clinical symptoms and improve clinical efficiency than that of WM alone for adults with non-severe CAP. Step-by-step results firstly indicated that XYP with WM was superior treatment regimen to reduce defervescence time, disappearance time of cough and disappearance time of lung rales. For safety, the mild adverse drug reactions occurred during the entire course of therapy and were self-limiting, further confirming the favorable safety profile of the TCMIs.

Enhancement of levopimaradiene production in Escherichia coli via engineering isopentenol utilization pathway

2026/1/26 16:03:19

Objective: Plant-derived terpenoid ginkgolides exhibit significant pharmacological efficacy, however, their extraction remains costly. Given that levopimaradiene is a key biosynthetic precursor to ginkgolides, its high-yield production via heterologous gene expression therefore establishes a critical foundation for scaling up their manufacture. This study primarily aims to enhance the yield of levopimaradiene in Escherichia coli (LB medium) by remodeling the unnatural isopentenol utilization pathway (IUP). Methods: Plasmid construction was driven by the mechanism of homologous recombination, which utilizes recombinase to facilitate the ligation process. The expression of kinase was carried out using type 7 (T7) promoter and isopropyl b-D-1-thiogalactopyranoside (IPTG) as the inducer. The quantification of levopimaradiene produced by E. coli was determined by comparison with a standard curve that we constructed. Results: Combining the selection of kinases, ribosome-binding site (RBS) screening, protein directed evolution and optimization of fermentation parameters, the production of levopimaradiene in E. coli was ultimately enhanced to 2691.3 mg/L, surpassing the highest reported titers of levopimaradiene with 6-fold in E. coli to date. Additionally, the engineered E. coli was designed to collaborate with farnesyl pyrophosphate (FPP) synthase and geranylfarnesyl pyrophosphate (GFPP) synthase to efficiently produce FPP and GFPP for sesquiterpene and sesterterpene synthesis. Conclusion: Our work showcases a combinatorial engineering strategy that employs an IUP-enhanced E. coli chassis for the microbial production of levopimaradiene, as well as other natural terpenoids.

Genome-wide identification and characterization of methyltransferase involved in benzylisoquinoline alkaloids biosynthesis from Menispermum dauricum

2026/1/26 16:03:19

Objective: Benzylisoquinoline alkaloids (BIAs) are valuable plant metabolites whose structural diversity largely depends on O-/N-methyltransferases (OMTs/NMTs). Although the CYP450-mediated backbone formation of BIAs was previously elucidated in Menispermum dauricum DC., the specific OMTs/NMTs responsible for their functional methylation remain unknown. This study aims to systematically identify and characterize these methyltransferases to define their roles in BIA biosynthesis. Methods: Combining genomic and biochemical approaches, a genome-wide identification of methyltransferase (MT) genes was conducted using the published M. dauricum genome. The candidate MTs were functionally evaluated through in vitro enzymatic assays employing diverse BIA substrates to determine their methylation specificities. The substrate-binding modes of MdOMT1 and MdOMT11 were predicted and compared by protein modeling and molecular docking. Results: Functional characterization of the 75 methyltransferases identified in the M. dauricum genome revealed four key enzymes (three OMTs and one NMT) that contribute to the diversification of BIA scaffolds through their distinct substrate specificities and positional preferences. Specifically, MdOMT1 preferentially catalyzed O-methylation at C7 position of 1-benzylisoquinolines (1-BIAs) and C2 position of tetrahydroprotoberberines. In contrast, MdOMT11 exhibited superior affinity and a strong preference for O-methylation at the C9 position of (S)-scoulerine. Meanwhile, MdNMT3 demonstrated effective Nmethylation activity toward both 1-BIAs and tetrahydroprotoberberines. Conclusion: This research elucidates the functional landscape of OMTs/NMTs in M. dauricum, revealing their crucial roles in BIA structural diversification. The newly identified enzymes provide valuable biocatalytic tools for synthetic biology approaches aimed at the sustainable and optimized production of highvalue BIAs.

Design and synthesis of butyrolactone V derivatives and its anti-inflammatory activity analysis

2026/1/26 16:03:19

Objective: To design and synthesize an amount of butyrolactone V derivatives, evaluate the antiinflammatory effects of all the derivatives, look for potential drugs that inhibit inflammatory bowel disease (IBD), and determine the structure-activity relationship (SAR). Methods: The butyrolactone V derivatives were synthesized with high yield by oxidation reaction, substitution reaction, and esterification reaction in sequence, and the production of nitric oxide was assessed in RAW264.7 cells treated with the lipopolysaccharide and the compounds. Then, the target compounds were studied for their activity in dextran sodium sulfate (DSS)-induced ulcerative colitis. Results: A total of three series of compounds encompassing 60 derivatives of the natural product butyrolactone V were designed and synthesized. The results showed that compounds 5p and 7e could alleviate the symptoms of DSS-induced colitis in mice, including alleviating diarrhea, inhibiting the reduction of colon length, and reducing tissue damage. The preliminary mechanism exploration indicated that compounds 5p and 7e could improve the symptoms of IBD in mice mainly by reducing the expression of chemokines and exerting anti-inflammatory effects. Conclusion: This study reports the synthesis and the derivatization of butyrolactone V and analysis on anti-inflammatory activity. The most effective compounds 5p and 7e have the potential to be further developed as drugs to treat IBD.

Precise subcellular organelle-targeted analyses of hepatotoxicity of Polygonum multiflorum

2026/1/26 16:03:19

Objective: Drug-induced liver injury (DILI) is an undesirable reaction caused by drugs, herbal medicines or supplements and may lead to acute liver failure. Polygoni Multiflori Radix (PMR, Heshouwu in Chinese) originated from the roots of Polygonum multiflorum is a popular traditional Chinese medicine (TCM) while potential hepatotoxicity limits its clinical application. The present study aims to elucidate the in-depth mechanism of PMR-induced organelle heterogeneity of hepatotoxicity. Methods: Network pharmacology and available TCM transcriptomics databases including Integrated Traditional Chinese Medicine (ITCM) and HERB databases were conducted to identify the active ingredients of PMR with the potent ability to injure organelles including microsome, mitochondria, endoplasmic reticulum (ER), Golgi apparatus (GA), and lysosome. Organelles were isolated and cultured with adenosine triphosphate (ATP)-supplemented system. Western blotting and particle size characterization techniques were further performed to clarify the organelle heterogeneity of PMR-induced hepatotoxicity. Results: Five representative organelles were isolated from mouse livers or hepatocytes and administrated with PMR-derived active components and monomers. The results of network pharmacology and virtual screening initially identified the components of PMR that may damage different organelles. By combining experimental verification, we found that five organelles studied in this research were the target organelles for flavonoid (FVN)’s affiliated compound quercetin (QC). Mitochondria were damaged mainly by kaempferol, anthraquinone (AQ) and its monomeric components. Meanwhile, QC and emodin showed effective toxicity on endoplasmic reticulum. For microsome, QC remained the most toxic monomer. For Golgi apparatus, trans-stilbene glycosides (trans-SG), AQ and emodin were the major toxic components in PMR. For lysosomes, total-SG, emodin and QC were the major toxic components in PMR. Conclusion: Collectively, our findings revealed the organelle heterogeneity of PMR-induced hepatotoxicity and identified quercetin as a potential toxic component in PMR. This study provides a novel conjunct strategy to screen and discover potential toxic components and shapes the understanding of toxicity warning and clinically safe use of herbal medicines such as PMR.

Chelidonine overcomes P-gp-mediated adriamycin resistance in MCF-7/ADR cells by inhibiting PDGFRb/PI3K/Akt pathway

2026/1/26 16:03:19

Objective: Chemoresistance represents a major obstacle in breast cancer (BC) treatment. Chelidonine could prevent various tumor cell types. However, the effect and mechanism of chelidonine against BC chemotherapy resistance have not been elucidated. This paper aimed to explore the effect and mechanism of chelidonine on BC chemoresistance. Methods: A CCK-8 assay, flow cytometry and fluorescence microscopy were applied to evaluate the resistance reversal effect of chelidonine on MCF-7/ADR cells. The signaling pathways by which chelidonine suppresses BC were predicted by network pharmacology and validated by Western blotting. The chemoresistant reversal mechanism of chelidonine was clarified using platelet-derived growth factor receptor-b (PDGFRb) silencing with small interfering RNA (siRNA), platelet-derived growth factor-BB (PDGF-BB) stimulation, Western blotting and real-time quantitative polymerase chain reaction (RTqPCR). Results: Chelidonine remarkably reversed adriamycin (ADR) resistance by decreasing P-glycoprotein (P-gp) expression and the efflux of ADR in MCF-7/ADR cells. Additionally, PDGFRb expression in MCF-7/ADR cells was markedly higher than that in MCF-7 cells (P < 0.01), and PDGFRb knockdown prevented P-gp expression and intracellular ADR accumulation. Network pharmacology identified phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) as a primary pathway of chelidonine-inhibiting BC, which was verified by the marked underexpression of phosphorylated kappa B inhibitor protein kinase (p-IKK), phosphorylated inhibitor of nuclear factor-jB (p-IKB), and nuclear factor-jB (NF-jB) and phosphatase and tensin homolog (PTEN) hyperexpression by chelidonine treatment (P < 0.01). Notably, PDGFRb silencing enhanced the inhibitory effect of chelidonine on the activation of the PI3K/Akt pathway. Moreover, chelidonine suppressed PDGF-BB stimulation of the PDGFRb/PI3K/Akt axis. Conclusion: These findings underscore the potential role of PDGFRb in regulating chemotherapy resistance in BC. Chelidonine could effectively overcome the resistance of MCF-7/ADR cells to ADR by targeting the PDGFRb/PI3K/Akt axis. Meanwhile, these findings highlight the potential of chelidonine as a promising natural chemoresistant agent for BC treatment.

Astragalus polysaccharides inhibit arsenic trioxide-induced BMSCs damage through inhibition of Jnk and p38 signaling pathways

2026/1/26 16:03:19

Objective: Arsenic trioxide (ATO) is a clinically effective anticancer agent used in the treatment of leukemia. However, it exerts adverse effects on non-tumor cells, including bone marrow mesenchymal stem cells (BMSCs). This study aims to investigate the protective role and molecular mechanism of traditional Chinese medicine Astragalus polysaccharides (APS) in mitigating ATO-induced apoptosis in BMSCs. Methods: BMSCs exposed to ATO (0.5 lmol/L) were treated with APS (20, 40, 100, and 200 lg/mL). Cell viability, proliferation, and migration were assessed by using MTT, EdU staining, Transwell, and scratch wound healing assays. Apoptosis was evaluated via TUNEL assay, Hoechst 33258 staining, and flow cytometry. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential were measured by using DCFH-DA and JC-1 staining. Apoptotic protein expression was analyzed by Western blotting. Results: ATO exposure significantly inhibited the proliferation and migration of BMSCs and induced apoptosis, while APS markedly attenuated the apoptosis of BMSCs induced by ATO, and significantly improved cell proliferation and migration (P < 0.01). Mechanistically, APS effectively reduced ATOinduced ROS (P < 0.01), while the protein expression of Bcl-2-associated X protein (Bax) and cleaved Caspase-3 was significantly decreased (P < 0.05), and the protein expression of Bcl-2 was significantly increased (P < 0.01). In addition, APS markedly decreased the protein expression of c-Jun N-terminal kinase (Jnk) and p38 in ATO-activated BMSCs (P < 0.05), and significantly decreased the protein expression of p16 and p53 (P < 0.01), and increased the protein expression of phosphorylated protein kinase B (p-Akt) and phosphorylated extracellular signal-regulated kinase (p-Erk) (P < 0.01, 0.05). Conclusion: Our study reveals that APS exert significant protective effects against ATO-induced apoptosis in BMSCs. The mechanisms involve suppressing ROS generation, maintaining mitochondrial membrane stability, enhancing cell viability, migration, and proliferation, as well as inhibiting Jnk and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways. The findings highlight potential molecular targets and novel strategies for the clinical prevention and treatment of ATO-related toxicity

Safflower yellow in Carthami Flos is responsible for Xuebijing Injectioninduced immediate hypersensitivity reaction through activating complement C3

2026/1/26 16:03:19

Objective: Xuebijing Injection (XBJI) is mainly used for treating sepsis in China, and even COVID-19 recently. This study aimed to clarify the molecular mechanism(s) and identify the potential ‘‘common culprit(s)” for XBJI-caused immediate hypersensitivity reaction (IHR) which is the main type of its adverse reactions. Methods: Antiserum against XBJI was prepared by intraperitoneal immunization in combination with aluminum adjuvant for five weeks. Antagonistic experiments were performed by using several antagonists against different mediators in Evans Blue leakage model. Propranolol-pretreated mice were used to determine the capacity of XBJI to trigger systemic IHR. Serum total IgE (tIgE) and mouse mast cell protease 1 (MCPT-1) levels, complement activation, and the levels of supernatant inflammatory mediators were determined by ELISAs. Lipopolysaccharide (LPS)-activated RAW264.7 macrophages were used for evaluating the anti-inflammatory activity of XBJI, while human mast cells (LAD2) were used for assessing the effect of XBJI on mast cell degranulation. Results: Continuous treatment (i.p.) with XBJI along with aluminum adjuvant did not elevate the levels of serum tIgE and MCPT-1. In vitro, XBJI could not directly cause the degranulation of LAD2 cells. It induced a robust Evans Blue leakage after the first injection in mouse paw. Mechanism study demonstrated that antagonists for histamine H1/H2 receptors and complement C3a receptor counteracted XBJI-induced IHR. XBJI also directly activated complement C3 in human serum. Through screening five herbs of XBJI and the constituents, only safflower yellow (SY) in Carthami Flos was able to induce IHR. The discolored-XBJI not only did not induce IHR locally and systemically, but also could suppressing the production of proinflammatory mediators in LPS-activated RAW264.7 macrophages. Conclusion: XBJI failed to induce immune IHR, but potently triggered non-immune IHR through direct activating complement C3 to provoke histamine release. SY in Carthami Flos was the underlying ‘‘common culprit” responsible for XBJI-caused IHR. The anti-inflammatory action of XBJI can be retained after decolorization. Our study provides a scientific basis for not only preventing and treating XBJI-caused IHR clinically, but also improving its production process.

Comprehensive genomic identification and functional analysis of bHLH transcription factors in Ganoderma lucidum

2026/1/26 16:03:19

Objective: The basic helix-loop-helix (bHLH) transcription factors (TFs) are pivotal in regulating fungal growth, development, and secondary metabolism. However, the knowledge about the Ganoderma lucidum bHLHs (GlbHLHs) in ganoderic acid (GA) biosynthesis of G. lucidum was limited. This study aimed to explore the functions of bHLH genes in ganoderic acid biosynthesis during G. lucidum growth development. Methods: First, the genome-wide identification of GlbHLHs was performed through Hidden Markov model searches and Two-way blast. Furthermore, through physicochemical properties, gene structure, and phylogenetic analysis, as well as combining the transcriptome and metabolome data from different developmental stages of G. lucidum, candidate GlbHLHs were screened. Subsequently, their regulatory roles in ganoderic acid biosynthesis were explored using yeast one-hybrid and dual luciferase reporter assays. Results: A total of 11 GlbHLH members were characterized in G. lucidum. The upstream promoter regions of these genes enriched hormones and abiotic stress responsive elements. Although individual ganoderic acid monomers demonstrated marked differences in accumulation patterns across specific growth phases and tissue types, overall, the total GA content was consistently higher in caps than in stipes throughout development. In addition, all GlbHLHs exhibited high expression in whole G. lucidum from the primordium to maturation stages. Among them, GlbHLH5 and GlbHLH7 showed the highest expression in any stage and highly correlated with key genes associated with GA pathway. Functional validation through dual-luciferase assays and yeast one-hybrid experiments had demonstrated that GlbHLH5 activated the P2 region of the lanosterol synthase promoter, while GlbHLH7 activated the promoters of squalene epoxidase and squalene synthase. Conclusion: Compared to plants, G. lucidum harbored a small number of bHLH members but all high expression in any stages. Additionally, GlbHLH5 and GlbHLH7 with the highest expression among GlbHLHs showed activation in regulating the biosynthesis of GA. These results provide a theoretical reference for further research on ganoderic acid regulation in G. lucidum, and thereby providing a molecular foundation for enhancing ganoderic acid yield to optimize the medicinal value of G. lucidum.

Morphological, physiological and biochemical changes and effects of polyamines on regulation of Anoectochilus roxburghii during flowering period

2026/1/26 16:03:20

Objective: Anoectochilus roxburghii is a valuable medicinal and ornamental plant. The aim of this study is to investigate the morphological and biochemical responses during the flower development stages of A. roxburghii, and to assess the effects of exogenous polyamines (PAs) on bud differentiation and metabolism, thereby providing a theoretical basis for understanding the flowering form and physiology of A. roxburghii. Methods: In this study, morphological and biochemical responses in flower development stages of A. roxburghii were investigated using paraffin sections and stereomicroscope. A. roxburghii was divided into five periods, including vegetative growth period, flower bud period, flowering period, late flowering period and fruiting period. During the flowering phase, specific biochemical parameters were measured, including soluble sugar content, superoxide dismutase (SOD) activity, soluble protein content, peroxidase (POD) activity, and catalase (CAT) activity. These measurements were conducted to understand the biochemical changes occurring within A. roxburghii during its flowering process. Furthermore, the effects of PAs on bud differentiation were examined. Additionally, the activities of S-adenosylmethionine decarboxylase (SAMDC) and polyamine oxidase (PAO), as well as the content of polyphenols, polysaccharides, and flavonoids in A. roxburghii, were measured after PA treatment to evaluate the metabolic changes induced by exogenous PAs. Results: During the flowering phase of A. roxburghii, soluble sugar content and SOD activity were steadily declining. Soluble protein content was initially increasing and then reducing, and POD and CAT activities showed opposite pattern. In addition, the effects of exogenous PAs on bud differentiation were investigated. Results showed that 3 mmol/L putrescine or 0.3 mmol/L spermidine significantly promoted the bud differentiation of A. roxburghii and advanced the flowering. The activities of SAMDC, PAO, and the content of polyphenols, polysaccharides and flavonoids in A. roxburghii significantly increased after PA treatment, demonstrating that exogenous PA can accelerate metabolism and improved the active ingredients content. Conclusion: The flower development of A. roxburghii was divided into five stages, with significant changes in soluble sugar, protein, POD, SOD, CAT, MDA, and PRO levels. Exogenous putrescine and spermidine enhanced bud differentiation and accelerated flowering, increasing SAMDC and PAO activities, suggesting accelerated PA metabolism. PAs also improved active component content. These findings provide a theoretical basis for studying flower morphology and PA-induced flowering regulation of A. roxburghii.

Portulaca oleracea alleviates CCl4-induced acute liver injury by regulating hepatic S100A8 and S100A9

Aruna Qian a,b, Lu Zhou a,b, Dongxu Shi a,b, Zongran Pang a,b,*, Binan Lu a,b,* — 2023/1/30 8:18:48

Objective: Acute liver injury (ALF) is a potential factor of many serious hepatopathies. Carbon tetrachloride (CCl4) is a possible environmental toxicant that can induce ALF. Portulaca oleracea (PO) is one of the most popular edible herbs and has several biological activities such as antioxidant, antimicrobial, antiinflammatory effects. We explored the significance of PO in regulating inflammatory function in animal models and cultured hepatocytes during liver damage caused by CCl4. Methods: The effect of PO on ALF was evaluated by CCl4-induced mice models in vivo. Hepatic levels of transaminase activities and inflammatory factors were examined. The gene and protein expression of S100A8 and S100A9 were measured by RT-PCR and Western blot analysis. Meanwhile, the efficacy of PO was certified by HepG2 cells in vitro. The transaminase activities, inflammatory factors, and the protein expression of S100A8 and S100A9 were also detected. Results: Animal tests showed that pretreatment with PO reduced the liver pathological tissue damage and the serum levels of ALT, AST, ALT and LDH, as well as reducing the pro-inflammatory cytokines (IL-1b, IL-6, TNF-a) secretion in CCl4-induced liver injury mice. Simultaneously, HepG2 cells pretreated with PO exhibited a significant decrease in the activities of ALT and AST. Moreover, PO resulted in a significant downregulation of the pro-inflammatory markers S100A8, S100A9 gene and protein expression on CCl4 induced acute liver injury was demonstrated entirely in vivo and vitro experiments. Conclusion: PO may down-regulate S100A8 and S100A9 and inhibit pro-inflammatory cytokines’ release, indicating a potential clinical effect for controlling the disease.