Determination of caffeine content in Ginkgo Folium by ultra high performance liquid chromatography-triple quadrupole mass spectrometry / 中国中药杂志

In this experiment, an ultra-high performance liquid chromatographytandem triple quadrupole mass spectrometry was established for the determination of caffeine in commercially available Ginkgo Folium. The samples were extracted by ultrasonic method with methanol, and separated on Waters CORTECS T3 column(2.1 mm×100 mm, 2.7 μm), with mobile phase of 0.1% formic acid solution-0.1% formic acid acetonitrile solution for gradient elution, at flow rate of 0.3 mL·min~(-1); column temperature of 30 ℃, and injection volume of 2 μL. Mass spectrometry was conducted at ESI~+ multiple reaction monitoring(MRM) mode; quantitative analysis was conducted with external standard method. The results showed that in the range of 0.099 6-9.96 ng·mL~(-1), there was a good linear relationship between the mass concentration of caffeine and the peak area, R~2=0.999; the average recovery was 84.51%, with RSD of 6.2%. The results of precision, repeatability and stability showed that the RSD was 5.1%, 5.9%, 7.2%, respectively. The content range of caffeine in 10 batches of Ginkgo Folium was 1.52-60.86 μg·kg~(-1). In conclusion, this method is accurate, reliable and reproducible, which provides a reference for the safety study of Ginkgo Folium.

The diversity of endophytic fungi in endangered medicinal material Daphnes Cortex as determined by high-throughput sequencing technology / 药学学报

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

Cloning，Bioinformatics and Expression Analysis of UGDH1 Gene in Glycyrrhiza uralensis / 中国实验方剂学杂志

Objective:To clone uridine diphosphate （UDP）-glucose dehydrogenase （<italic>UGDH</italic>） gene of <italic>Glycyrrhiza uralensis</italic> and analyze its bioinformatics and expression. Method:Total RNA was extracted from roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， the complementary deoxyribonucleic acid （cDNA） sequence of <italic>GuUGDH</italic>1 gene （Gu was short for <italic>G. uralensis</italic>） was cloned by reverse transcription polymerase chain reaction （RT-PCR）， then sequencing and bioinformatic analysis were performed， and the specificity of the tissue was analyzed by real-time fluorescence quantitative PCR （Real-time PCR）. Result:The open reading frame（ORF）of <italic>GuUGDH</italic>1 gene was 1 443 bp in length and encoded 480 amino acid residues （GenBank accession number of MT968993）. Bioinformatics analysis showed that GuUGDH1 was a stable acidic hydrophilic protein with a relative molecular weight of 53.056 kDa， an isoelectric point of 5.89， no signal peptide and no transmembrane helix， and all of them were outside the membrane. There were three typical conserved domains， which belonged to the UDP-glucose/guanosine diphosphate （GDP）-mannose dehydrogenase family. Phylogenetic analysis showed that the <italic>GuUGDH</italic>1 gene was closely related to <italic>Glycine max</italic> and <italic>Spatholobus suberectus</italic>. The results of Real-time PCR showed that the expression of <italic>GuUGDH</italic>1 gene could be detected in the roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， and the expression level in the roots was significantly higher than that in the stems and leaves. Conclusion:In this study， the <italic>UGDH</italic>1 gene of <italic>G. uralensis</italic> was cloned and its protein sequence characteristics were systematically analyzed， which can provide theoretical basis for further research on the catalytic function of UGDH1 protein.

CCT4 suppression inhibits tumor growth in hepatocellular carcinoma by interacting with Cdc20 / 中华医学杂志(英文版)

BACKGROUND@#The chaperonin containing t-complex (CCT) proteins play an important role in cell cycle-related protein degradation in yeast and mammals. The role of the chaperonin containing t-complex 4 (CCT4), one subtype of CCT proteins, in the progress of hepatocellular carcinoma (HCC) was not fully elucidated. Here, we aimed to explore the mechanisms of CCT4 in HCC.@*METHODS@#In this study, we used the UALCAN platform to analyze the relationship between CCT4 and HCC, and the association of CCT4 with the overall survival (OS) of HCC patients was also analyzed. CCT4 expression in HCC tumor tissues and normal tissues was also determined by western blot (WB) assay. Lentivirus vector was used to knock down the CCT4 expression, and quantitative polymerase chain reaction and WB were used to determine the level of CCT4 in HCC cell lines. Cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assays were used to detect the cell proliferation, and flow cytometry (FCM) was performed to evaluate the effect of CCT4 on the apoptosis of HCC cells. Co-immunoprecipitation (co-IP) assay and WB were used to explore the mechanisms of CCT4 regulating the growth of HCC. Data were calculated from at least three replicate experiments and expressed as mean ± standard deviation. Student's t test, paired t test, and Kaplan-Meier analysis were used to compare across different groups.@*RESULTS@#We found CCT4 was upregulated in HCC tissues compared with normal tissues, and its high expression was associated with poor prognosis (P < 0.001). CCT4 was significantly increased in HCC tumor tissues compared with normal tissues (0.98 ± 0.12 vs. 0.23 ± 0.05, t = 7.73, P < 0.001). After being transfected with CCT4 short-hairpin RNA (shRNA), CCT4 was decreased in mRNA level and protein level in both Huh7 (mRNA level: 0.41 ± 0.07 vs. 1.01 ± 0.11, t = 8.09, P = 0.001; protein level: 0.61 ± 0.03 vs. 0.93 ± 0.07, t = 7.19, P = 0.002) and Hep3b cells (mRNA level: 0.55 ± 0.11 vs. 1.04 ± 0.15, t = 4.51, P = 0.011; protein level: 0.64 ± 0.10 vs. 0.95 ± 0.08, t = 4.32, P = 0.012). CCK8 assay indicated that CCT4 knockdown inhibited cell proliferation in both Huh7 (OD value of 3 days: 0.60 ± 0.14 vs. 0.97 ± 0.16, t = 3.13, P = 0.036; OD value of 4 days: 1.03 ± 0.07 vs. 1.50 ± 0.12, t = 5.97, P = 0.004) and Hep3b (OD value of 3 days: 0.69 ± 0.14 vs. 1.10 ± 0.11, t = 3.91, P = 0.017; OD value of 4 days: 1.12 ± 0.12 vs. 1.48 ± 0.13, t = 3.55, P = 0.024) cells. EdU assay showed that CCT4 knockdown inhibited the cell proliferation in both Huh7 (EdU positive rate: [31.25 ± 3.41]% vs. [58.72 ± 3.78]%, t = 9.34, P = 0.001) and Hep3b cells (EdU positive rate: [44.13 ± 7.02]% vs. [61.79 ± 3.96]%, t = 3.79, P = 0.019). FCM assay suggested that CCT4 knockdown induced apoptosis in HCC cells (apoptosis rate of Huh7: [9.10 ± 0.80]% vs. [3.66 ± 0.64]%, t = -9.18, P = 0.001; apoptosis rate of Hep3b: [6.69 ± 0.72]% vs. [4.20 ± 0.86]%, t = -3.84, P = 0.018). We also found that CCT4 could regulate anaphase-promoting complex (APC)Cdc20 activity via interacting with Cdc20. Furthermore, CCT4 knockdown induced securin (0.65 ± 0.06 vs. 0.44 ± 0.05, t = -4.69, P = 0.009) and B-cell lymphoma-2 (Bcl-2) interacting mediator of cell death (Bim; 0.96 ± 0.06 vs. 0.61 ± 0.09, t = -5.65, P = 0.005) accumulation. The upregulation of securin inhibited cell growth by downregulating cyclin D1 (0.65 ± 0.05 vs. 1.04 ± 0.07, t = 8.12, P = 0.001), and the accumulation of Bim inhibited Bcl-2 (0.77 ± 0.04 vs. 0.87 ± 0.04, t = 3.00, P = 0.040) and activated caspase 9 (caspase 9: 0.77 ± 0.04 vs. 0.84 ± 0.05, t = 1.81, P = 0.145; cleaved caspase 9: 0.64 ± 0.06 vs. 0.16 ± 0.07, t = 1.81, P = 0.001), which led to elevated apoptosis.@*CONCLUSIONS@#Overall, these results showed that CCT4 played an important role in HCC pathogenesis through, at least partly, interacting with Cdc20.

Polyphenols from Securidaca inappendiculata alleviated acute lung injury in rats by inhibiting oxidative stress sensitive pathways / 中草药·英文版

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

Study on Inhibitory Effect of Volatile Oils in 16 Plants on Aspergillus flavus of Platycladi Semen / 中国实验方剂学杂志

Objective::To select volatile oils from 16 species of plants (Cymbopogon citratus, Pelargonium graveolens, Pinus tabulieformis, Litsea cubeba, Mentha haplocalyx, Zingiber officinale, Syzygium aromaticum, Curcuma longa, Zanthoxylum bungeanum, Cinnamomum cassia, Ocimum basilicum, Rosmarinus officinalis, Zanthoxylum schinifolium, Zanthoxylum armatum, Illicium verum, Myristica fragrans) that have good inhibitory effect on the growth of Aspergillus flavus. Method::Aspergillus flavus was isolated from the surface of Platycladi Semen medicinal materials by plate culture method. The volatile oils of 16 plants were extracted by steam distillation. The colony diameter of Aspergillus flavus was determined by fumigation of filter paper, and the effect of volatile oils on the growth of Aspergillus flavus was systematically studied. Result::Aspergillus flavus was successfully isolated from Platycladi Semen by means of morphological, microscopic and DNA barcoding identification methods, the bacteriostatic rates of the above 16 kinds of volatile oils against Aspergillus flavus were 2.93%, 0.05%, 0.37%, 76.07%, 0.34%, 0.15%, 50.05%, 8.51%, 1.43%, 58.20%, 0.07%, 2.60%, 8.73%, 100.00%, 52.62%, 0.07%, respectively. Conclusion::The volatile oils of 16 plants all have different degrees of antibacterial activities for Aspergillus flavus, and volatile oils of Zanthoxylum armatum, Litsea cubeba and Cinnamomum cassia have good inhibitory effect. This study can provide a theoretical basis for the prevention and control of Aspergillus flavus in the growth and storage of Platycladi Semen, and provide a basis for further research on plant volatile oil as bacteriostatic agents in the storage process of traditional Chinese medicine.

Isolation and Elucidation of a Novel Isoquinoline Alkaloid from Corydalis hendersonii / 中国实验方剂学杂志

As part of systematic research of Corydalis hendersonii,a typical traditional Tibetan herbal medicine with clearing heat,relieving pain,and lowering blood pressure effects,a novel isoquinoline alkaloid,named hendersine G was isolated from the ethanol extract of the whole plant by various chromatographic techniques,including silica gel column,reverse phase column (ODS),Sephadex LH-20,and semi-preparative HPLC.Its structure was elucidated by MS,NMR and other spectroscopic data analysis.Hendersine G can be regarded as a condensation product of a tetrahydroberberine and a succinic acid,however,its absolute configuration has not been determined due to its structural complexity and less obtained amount.This present study provides an inspiration for further exploration of novel molecules from C.hendersonii.

Research Progress on Prevention and Control Measures of Aflatoxin in Chinese Medicinal Materials / 中国实验方剂学杂志

At present,traditional Chinese medicine(TCM) has attracted more and more attention from the international community.The demand for TCM is increasing in the world.The hidden dangers of potential quality and safety of TCM are also becoming increasingly prominent.Aflatoxin contamination has become one of the important factors affecting the safety of Chinese herbal medicines,and it will fundamentally affect human health and life safety.A variety of methods are used to reduce aflatoxins,however,there are few suitable methods that can be widely used in the cost-effective and large-scale promotion of Chinese herbal medicines.Therefore,it is of great significance to continue to study measures to solve the pollution problems of Aspergillus flavus and its toxins.This article summarizes the hazards and contamination status of aflatoxin,the prevention and control of the growth of A. flavus, and the measures for reducing aflatoxin,and looks ahead to the future prevention and control of A. flavus and its toxins,aiming at providing ideas for the pollution problem of A. flavus and its toxin,to ensure the quality of Chinese herbal medicines,so as to ensure clinical safety medication.

Reflections on Relationship Between Pathogenesis of Endometriosis and Intestinal Flora / 中国实验方剂学杂志

Endometriosis is a multiple disease that afflicts the health of women at childbearing age,and its incidence rate has been increasing year by year,furthermore,there has been a trend to be younger.At present,the pathogenesis of endometriosis has been not expounded completely,its cure rate is not high with high recurrence rate.In recent years,studies have shown that the human is a commensal body composed of a large number of microorganisms,and especially the microorganisms in the intestinal are closely related to the health of the body.Based on the previous studies on endometriosis,this paper proposes that its pathogenesis may be related to intestinal microbiological disorder,and aims to provide new ideas for the treatment of endometriosis.

Analgesic and sedative effects of Mongolian medicine Syringa pinnatifolia / 中国中药杂志

The peeled root,stem or twig of Syringa pinnatifolia is a representative Mongolian folk medicine with the effects of antidepression and pain relief. It has been used for the treatments of heart tingling,heart palpitations,upset,insomnia and other symptoms. Inspired by Mongolian medical theory and clinical practices,this study evaluated the analgesic effect of S. pinnatifolia ethanol extract( T) through three analgesic models including acetic acid writhing test,formalin test,and hot plate test,and the sedative effect of T was evaluated by locomotor activity and synergistic sleeping experiments,and furthermore the effects of T on the GABAergic nervous system were investigated by ELISA,immunohistochemistry,Western blot,and PCR methods. The results showed that T can significantly reduce the number of writhing,the time of paw licking and extend the thermal threshold of mice,suggesting the analgesic effect of T.T also can indicate its sedative effect by reducing the number of activities,decreasing latency of sleeping and extending sleeping time of mice. ELISA results showed that T can increase the content of GABA/Glu in rat cortex,hippocampus,and hypothalamus,and the most significant increase in hypothalamus. The immunohistochemistry and Western blot results showed that T can up-regulate the expression of GAD67 protein in hypothalamus,and the PCR results showed that T can up-regulate the expression of GABAA Rα1,α2,α3,α5,β1-3,γ1-3 genes,suggesting a sedative effect through the GABAergic nervous system. In conclusion,this study shed insight into the theoretical basis and clinical application of S. pinnatifolia,and also provides inspiration for subsequent development and application.

Diversity of microbiota composition of traditional Chinese medicine of Faeces Trogopterori from Shangluo, Shanxi province by sequence of the bacterial 16S rRNA gene / 药学学报

To obtain the microbial composition of traditional Chinese medicine of Faeces Trogopterori, ten samples were collected from the imitate wildness farmland in Shangluo City, Shaanxi Province. In this study, 16S rRNA gene was used as molecular marker to explore the microbiome and the sequences were analyzed by Usearch analysis platform. The COG and KEGG database is used to predict and analyze the function of the flora. A great number of 285 218 high quality clean reads with a length of 400-450 bp were obtained from 10 samples. Bacterial species detected in these samples covered 8 phyla, 25 families, 75 genera and 120 species. The dominant phylum microbial communities in these samples were Firmicutes (87.68% ± 2.68%) and the Bacteroidetes (7.62% ± 3.74%), all samples showed a high microbial diversity, the predicted functional metagenome was heavily involved in energy metabolism. This study provided that the beneficial bacteria in Faeces Trogopterori may be one of its active ingredients, and no pathogens are detected in the sample.

Study on the surface fungus diversity of commercially Platycladi semen based on high-throughput sequencing technology / 药学学报

The structural composition of the surface fungal community of commercially Platycladi semen was analyzed to reveal the surface fungal biodiversity and structural differences. Platycladi semen was collected from Henan, Shandong and Hong Kong, their DNA was extracted, ITS fragments in DNA were amplified by PCR. Miseq was sequenced on Illumina Hiseq 2500 platform after the PCR products were qualified for quality inspection. The sequence OTU cluster was obtained and bioinformatics analysis was carried out. Microbial communities were not observed in the eyes of the Platycladi semen in the three regions. Sequencing results showed that the surface microbial community had high biodiversity, but there were significant differences in species composition. Seven samples o Platycladi semen obtained 345 947 valid sequences, which were divided into 267 OTUs, 3 phylums. 18 classes, 40 orders, 82 families, 120 genus, 191 species fungi. At the genus level, Aspergillus is the dominant species, accounting for the highest proportion, reaching (93.36 ± 6.01)%. Seven samples were contaminated by Aspergillus flavus, and the pollution levels were 14.58%, 15.98%, 17.64%, 16.44%, 0.97%, 23.39% and 18.86%. Except sample No. 5, Aspergillus cibarius was the most abundant, the other six samples were Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus as the core microflora. By analyzing the diversity of fungi distribution in different habitats, we can fully understand the fungi on the surface of Platycladi semen, lay a foundation for early risk warning of Aspergillus flavus contamination and its aflatoxin contamination, and provide a theoretical basis for the quality and safety of Platycladi semen.

Diversity and isolation parameters of endophytes from Glycyrrhiza uralensis of different habitats / 药学学报

In order to determine the differences in structure and optimum isolation conditions of Glycyrrhiza uralensis endophytes from different habitats, plate-separation method was used to identify endophytes in G. uralensis from Gansu, Ningxia, Inner Mongolia, Xinjiang, and Beijing. The isolation parameters were defined by investigating various concentrations and sterilization time of NaClO solution. The strains were identified by morphological and molecular biological methods. The results showed that 5% NaClO solution and sterilization time of 5 min were the optimal surface sterilization conditions. Among 129 strains of G. uralensis from 5 producing areas, 438 strains of endophytic fungi were isolated and belonged to 5 orders, 7 genera, and 11 species. Among them, 4 taxa were firstly isolated from the licorice in China. Fusarium was a common genus among the 5 regions. There were differences in the composition and structure of the endophytic fungi of G. uralensis from different habitats. Diversity analysis showed that the endophytic fungi diversity in Gansu was the highest and that of Beijing was the lowest. The comprehensive analyses indicated that the endophytic fungi of G. uralensis are diverse, and there were differences among the number, composition and population of endophytic fungi in five producing areas of Gansu, Ningxia, Inner Mongolia, Xinjiang and Beijing.

Effect of RNA interference on WD101 gene of Schistosoma japonicum

Objective:To study the effect of RNA interference (RNAi) on WD101 gene and its effect on the expression of WD101 mRNA and protein in Schistosoma japonicum.Methods:Double-stranded RNA (dsRNA) WD101 gene and control gene (lacZ) were generated by in vitro transcription and transfected into mechanically transformed schistosomula. The total RNA and protein were isolated simultaneously using TRIzol reagent. The expression levels of mRNA and the protein were determined by quantitative real-time PCR (qPCR) and Western blotting, respectively. After injected dsRNA-electroporated schistosomula into BALB/c mouse six weeks, the male and female reproductive organs were observed and measured under the confocal laser scanning microscope.Results:After 1, 3 and 5 d of RNAi, WD101 mRNA level was decreased by 15%, 39%, and 58% in experiment group compared to that in control group; meanwhile, WD101 protein level was decreased by 11%, 28%, and 43% in experiment group compared to that in control group. There were significantly more sperms in testicular lobes in experiment group than that in control group, while there were no significant differences in terms of ovary and vitelline glands between two groups.Conclusions:The dsWD101-RNAi can effectively induce suppression of WD101 gene expression at both mRNA and protein levels. WD101 gene might be a reproduction-related gene in Schistosoma japonicum.

Chemical and pharmacological progress on usnic acid and its derivatives / 中国中药杂志

Usnic acid and its derivatives, a group of organic molecules with great importance, are characteristic to lichens, possessing pharmacological activities such as anti-virus, anti-bacteria, anti-humor, anti-inflammatory, analgesic, and anaesthetic effects. Many of them have been widely used as medicine, but also bring side effects such as dermatitis and liver damages. In the past decades, great efforts by isolation, organic synthesis, and structure modification methods were put on discovery of UA derivatives with higher biological activities or less side effects. This paper describes herein the most progress on natural sources, isolation and structure elucidation, structural characteristics, synthesis and modification results, pharmacological activities and toxicities of UA and its derivatives, hopefully to provide valuable reference for further research.

Rapid Determination of γ-Hydroxybutyric Acid in Beverage and Urine by Direct Analysis in Real-time Mass Spectrometry / 分析化学

A method for rapid determination of γ-hydroxybutyric acid (GHB) in beverages (water, sodas, beer) and urine was established by direct analysis real-time mass spectrometry (DART-MS). Samples were analyzed directly after dilution with mixture of methanol and water(1:1,V/V). Instrument parameter settings were optimized to obtain the sensitive and accurate determination of GHB. At the sample introduction speed of 0.5 mm/s, high intensity of[M-H]- ions for GHB were observed in the negative ion and selection ion monitoring mode by utilization of high purity helium gas at 350℃. For different samples of water,sodas,beer and urine,the limits of detection (LODs) (S/N=3) were in the range of 1-2 μg/mL, while the limits of quantification (LOQs) (S/N=10) were in the range of 3-5 μg/mL. The linear correlation coefficients of the standard curves with different sample matrixes were between 0.9899 and 0.9980. The recoveries were in the range of 80.8%-115.2% with the relative standard deviations of 1.9%-12.8%. With its rapid analysis and simple pretreatment steps,the method is expected to have a strong advantage in the rapid screening analysis of large quantities of beverage and urine.

Effect of RNA interference on WD101 gene of Schistosoma japonicum

Objective: To study the effect of RNA interference (RNAi) on WD101 gene and its effect on the expression of WD101 mRNA and protein in Schistosoma japonicum. Methods: Double-stranded RNA (dsRNA) WD101 gene and control gene (lacZ) were generated by in vitro transcription and transfected into mechanically transformed schistosomula. The total RNA and protein were isolated simultaneously using TRIzol reagent. The expression levels of mRNA and the protein were determined by quantitative real-time PCR (qPCR) and Western blotting, respectively. After injected dsRNA-electroporated schistosomula into BALB/c mouse six weeks, the male and female reproductive organs were observed and measured under the confocal laser scanning microscope. Results: After 1, 3 and 5 d of RNAi, WD101 mRNA level was decreased by 15%, 39%, and 58% in experiment group compared to that in control group; meanwhile, WD101 protein level was decreased by 11%, 28%, and 43% in experiment group compared to that in control group. There were significantly more sperms in testicular lobes in experiment group than that in control group, while there were no significant differences in terms of ovary and vitelline glands between two groups. Conclusions: The dsWD101-RNAi can effectively induce suppression of WD101 gene expression at both mRNA and protein levels. WD101 gene might be a reproduction-related gene in Schistosoma japonicum.

A fluorescence polarization immunoassay for the detection of aflatoxins in herbal teas / 药学学报

A rapid fluorescence polarization immunoassay (FPIA) has been developed for the determi-nation of aflatoxins in samples of naturally-contaminated herbal teas. The tracers were synthesized by chemical method and determined by thin layer chromatography (TLC) and mass spectroscopy (MS). Fluorescence polarization was evaluated by the detection of polarized light. The results showed that the limit of detection (LOD) of FPIA for aflatoxins was 20 ng·mL-1, the IC50 was 371.80 ng·mL-1, and the linear range of the developed FPIA was 92.76-252.32 ng·mL-1. Compared with conventional HPLC methods, the FPIA developed in this study has the advantages of short analysis time and low cost. This method may be suitable for high- throughput screening of aflatoxins in herbal teas.

Effect of exogenous brassinolide on morphological characters and contents of seven chemical constituents of Glycyrrhiza uralensis / 中国中药杂志

The transplants of the two-year-old Glycyrrhiza uralensis were subjected to four concentration of brassinolide (BR 0.1, 0.4, 0.7, 1.0 mg•L⁻¹) in July. The morphological characters ( plant height, stem diameter, nodes number, internode length and root length , root thick, root fresh weight and root dry weight ) were measured and seven kinds of chemical constituents (glycyrrhizic acid, liquiritin, isoliquiritin, liquiritigenin, isoliquiritigenin, liquiritin apioside, isoliquiritin apioside) were determined by HPLC with the aim of increasing sinter output and improving quality of G. uralensis. Then the long-term dynamic changes of these morphological characters and chemical compositions' content were analyzed. The results showed that morphological characters of plant height, stem diameter, root length , root thick, root fresh weight and root dry weight increased remarkably with the 0.7 mg•L⁻¹ BR stimulating 2 months later,the increase rates were： 15.09%,6.15%,16.52%,8.46%,21.90%,29.41%, respectively. The content of glycyrrhizic acid, liquiritin, isoliquiritin, liquiritigenin, liquiritin apioside, isoliquiritin apioside were increased 20.16%,45.31%,53.56%,27.66%,23.54%,8.46% with the 0.7 mg•L⁻¹ BR stimulating 2 months later. The best effects were achieved in 2 months after brassinolide stimulating. The conclusions prove that morphological characters and the main chemical constituents accumulation of G. uralensis could be effected by exogenous BR stimulation in certain case.

Resource situation investigation and analysis about endangered Chinese herbal medicine Daphnes Cortex / 中国中药杂志

In order to figure out the status and distribution of the wild and cultivated resources of traditional Chinese medicine Daphnes Cortex, its suitable habitat and endangering factors were analyzed to provide the basis for its rational use, protection and cultivation.Our research group tooka resources survey in Shanxi, Gansu, Sichuan and Qinghai provinces, which include 23 counties. Investigation and sampling investigation combined with interview were carried out. The total reserve of resources was estimated through route-quadrat method in combination with the vegetation and soil-type map area method. The results indicated that there was no obvious change between the present distribution ranges of the wild Daphnes Cortex and its historical records, but the density of the population has undergone major changes. The wild reserves resources has declined seriously, even on the verge of exhaustion in some regions. According to the survey results, the current total reserve of the wild Daphnes Cortex in the four provinces was no more than 600 tons. Simultaneously, we only found the cultivated resource in a mountain at an altitude of about 2 800 m in Kang county of Gansu province, which cropping scope was about 33 000 m². The cultivated resource can't provide medicinal products at present, because their growing period is too short to have curative effect. Destructive excavation and the longer growth cycle result in a sharp decline of the wild resources reserves, even to the point of extinction. Artificial cultivation of product will become the main source of medicinal resources in the future. Therefore, we must protect its suitable habitat, formulate rational harvesting policy, strengthen the supervision of government departments, collect and establish the germplasm nursery and seed bank. On the basis, we must carry out studies into seed-selecting and breeding as well as rapid propagation and growth technology at once.