Volume 9,Issue 3,2017 Table of Contents

  • Display Type:
  • Text List
  • Abstract List
  • 1  Looking forward to the Future to Meet the Challenge: Research for Rhizosphere Microbes and Mocrobiome of Medicinal Plants
    Chang-xiao Liu
    2017, 9(3):197-198. DOI: 10.1016/S1674-6384(17)60096-0
    [Abstract](477) [HTML](0) [PDF 0.00 Byte](5)
    Abstract:
    2  Rhizosphere Microbiota and Microbiome of Medicinal Plants: From Molecular Biology to Omics Approaches
    Da-cheng Hao Pei-gen Xiao
    2017, 9(3):199-217. DOI: 10.1016/S1674-6384(17)60097-2
    [Abstract](1079) [HTML](0) [PDF 0.00 Byte](31)
    Abstract:
    The rhizosphere is a narrow region of soil that is directly influenced by roots and associated soil microorganisms. Research on rhizosphere microbes of various medicinal plants is essential for microbial ecology, applied microbiology and industrial biotechnology with regard to the sustainable utilization of Chinese medicinal resources. However, the inability of culturing most rhizosphere microorganisms (around 99%) in the laboratory obviates the research progress. In recent years, there is enormous advances in applying non-culturing techniques based on molecular biology and omics to the study of rhizosphere microbial diversity and plant-microbe interactions. DGGE, T-RFLP, ARDRA, DNA cloning and Sanger sequencing are still useful in the rhizosphere studies, while various omics tools, such as FISH, SIP, microarray, next generation sequencing (NGS), etc., evolve quickly to provide more comprehensive understanding of the rhizosphere microbiota and microbiome. Flexible applications of NGS technologies are here exemplified, e.g., amplicon sequencing, metagenomic sequencing, whole genome sequencing, and transcriptome sequencing, which address the biology and biotechnology potentials of the rhizosphere microbiome of medicinal plants. This review discusses recent findings and future challenges in the study of rhizosphere microbes, highlighting medicinal plant rhizosphere study, evolution of research methods, and innovative combinations of novel high-throughput techniques. The top-down approaches such as metagenomics and bottom-up approaches targeting individual species or strains should be integrated and combined with modeling approaches to afford a wide-ranging understanding of the microbial community as a whole.
    3  Authenticity Survey of Cuscutae Semen on Markets Using DNA Barcoding
    Zi-tong Gao Li-li Wang Xiao-yue Wang Yang Liu Jian-ping Han
    2017, 9(3):218-225. DOI: 10.1016/S1674-6384(17)60098-4
    [Abstract](913) [HTML](0) [PDF 0.00 Byte](14)
    Abstract:
    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.
    4  Validation of Reliable Reference Genes for Accurate Normalization in RT-qPCR Analysis of Codonopsis pilosula
    Ling-ya Cao Xiao-xiao Li Dong Wang Hai-feng Sun Jian-ping Gao
    2017, 9(3):226-235. DOI: 10.1016/S1674-6384(17)60099-6
    [Abstract](782) [HTML](0) [PDF 0.00 Byte](11)
    Abstract:
    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.
    5  Sensitive Analysis and Pharmacokinetic Study of Berberrubine Using LC-MS/MS
    Yu-qing Zhao Na Yang Fei Fei Run-bin Sun Si-qi Feng Jun He Jing-qiu Huang Yue Xie Ji-ye Aa Guang-ji Wang
    2017, 9(3):236-249. DOI: 10.1016/S1674-6384(17)60100-X
    [Abstract](985) [HTML](0) [PDF 0.00 Byte](11)
    Abstract:
    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.
    6  Rapid Determination of Total Content of Five Major Anthraquinones in Rhei Radix et Rhizoma by NIR Spectroscopy
    Hao Zhan Jing Fang Hong-wei Wu Hong-jun Yang Hua Li Zhu-ju Wang Bin Yang Li-ying Tang Mei-hong Fu
    2017, 9(3):250-257. DOI: 10.1016/S1674-6384(17)60101-1
    [Abstract](837) [HTML](0) [PDF 0.00 Byte](12)
    Abstract:
    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.
    7  UFLC-DAD-ESI-IT-TOFMSn Analysis on Biotransformation of Tongmai Formula Incubated with Human Intestinal Bacteria
    Shuai Wu Qi-le Zhou Xiu-wei Yang
    2017, 9(3):258-266. DOI: 10.1016/S1674-6384(17)60102-3
    [Abstract](746) [HTML](0) [PDF 0.00 Byte](9)
    Abstract:
    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.
    8  Simultaneous Determination of Saponins in Dripping Pills Made from Astragali Radix and Panax notoginseng by UPLC-ELSD
    Jing-jing Peng Dong-xiang Li Jing-yi Huang Ling Tong Bo-yang Yu
    2017, 9(3):267-274. DOI: 10.1016/S1674-6384(17)60103-5
    [Abstract](734) [HTML](0) [PDF 0.00 Byte](9)
    Abstract:
    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.
    9  Comprehensive Evaluation on Effect of IMPX977 on Expression of Methyl-CpG-binding Protein 2 in Rats
    Ya Hu Liang Le Li-xin Qi Ying Zhao Hui Fu Chen Duan Xiao-ying Wang Ke-ping Hu
    2017, 9(3):275-281. DOI: 10.1016/S1674-6384(17)60104-7
    [Abstract](830) [HTML](0) [PDF 0.00 Byte](12)
    Abstract:
    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.
    10  Protective Effects of Hydroxysafflor Yellow A against Oxidative Damage of β-Mercaptoethanol During Neural Differentiation of Mesenchymal Stem Cells
    Xiao-qing Song Li-ning Su Hui-ping Wei Ying-hui Liu Hai-feng Yin
    2017, 9(3):282-288. DOI: 10.1016/S1674-6384(17)60105-9
    [Abstract](624) [HTML](0) [PDF 0.00 Byte](6)
    Abstract:
    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.
    11  Phytochemical, Anti-oxidant, Larvicidal, and Antimicrobial Activities of Castor (Ricinus communis) Synthesized Silver Nanoparticles
    Namita Soni Ramesh C. Dhiman
    2017, 9(3):289-294. DOI: 10.1016/S1674-6384(17)60106-0
    [Abstract](842) [HTML](0) [PDF 0.00 Byte](13)
    Abstract:
    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.
    12  Inhibition of Cytochrome P450 by Nomilin and Obacunone and Potential Mechanism in Human Liver Microsomes
    Yao-wen Fan Yun-long Chen Jun-xiu Chen Fang-liang Zhang Gregory Ondieki Ya-zhuo Li Xin He
    2017, 9(3):295-298. DOI: 10.1016/S1674-6384(17)60107-2
    [Abstract](417) [HTML](0) [PDF 0.00 Byte](12)
    Abstract:
    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.

    Current Issue


    Volume , No.

    Table of Contents

    Archive

    Volume

    Issue

    Most Read

    Most Cited

    Most Downloaded

    WeChat

    Mobile website