Extracts of Poria cocos polysaccharides improves alcoholic liver disease in mice via CYP2E1 and NF-κB inflammatory pathways / 中国中药杂志

The present study investigated the effect of extract of Poria cocos polysaccharides(PCP) on cytochrome P450 2 E1(CYP2 E1) and nuclear factor κB(NF-κB) inflammatory signaling pathways in alcoholic liver disease(ALD) mice and explored its protective effect and mechanism. Sixty male C57 BL/6 N mice of SPF grade were randomly divided into a control group, a model group, a positive drug group(bifendate, 200 mg·kg~(-1)), and high-(200 mg·kg~(-1)) and low-dose(50 mg·kg~(-1)) PCP groups. Gao-binge mo-del was induced and the mice in each group were treated correspondingly. Liver morphological and pathological changes were observed and organ index was calculated. Serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected. Malondialdehyde(MDA) and superoxide dismutase(SOD) in liver tissues were detected by assay kits. The levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) were detected by ELISA. The activation of macrophages was observed by immunofluorescence staining and protein expression of CYP2 E1, Toll-like receptor 4(TLR4), NF-κB p65, and phosphorylated NF-κB p65(p-NF-κB p65) were analyzed by Western blot. The ALD model was properly induced. Compared with the model group, the PCP groups significantly improved the pathological injury of liver tissues. Immunofluorescence staining revealed that compared with the model group, the groups with drug intervention showed decreased macrophages in liver tissues. Additionally, the PCP groups showed reduced ALT, AST, MDA, IL-6, and TNF-α(P<0.05), and potentiated activity of SOD(P<0.01). PCP extract has the protective effect against alcoholic liver injury in mice, and the underlying mechanism may be related to the regulation of the expression of CYP2 E1 and inhibition of TLR4/NF-κB inflammatory signaling pathway to reduce oxidative stress and inflammatory injury, thereby inhibiting the development of ALD.

Study on the mechanism of Wuzi-Yanzong-Wan-medicated serum interfering with the mitochondrial permeability transition pore in the GC-2 cell induced by atractyloside / 中国天然药物

Wuzi-Yanzong-Wan (WZYZW) is a classic prescription for male infertility. Our previous investigation has demonstrated that it can inhibit sperm apoptosis via affecting mitochondria, but the underlying mechanisms are unclear. The purpose of the present study was to explore the actions of WZYZW on mitochondrial permeability transition pore (mPTP) in mouse spermatocyte cell line (GC-2 cells) opened by atractyloside (ATR). At first, WZYZW-medicated serum was prepared from rats following oral administration of WZYZW for 7 days. GC-2 cells were divided into control group, model group, positive group, as well as 5%, 10%, 15% WZYZW-medicated serum group. Cyclosporine A (CsA) was used as a positive control. 50 μmol·L-1 ATR was added after drugs incubation. Cell viability was assessed using CCK-8. Apoptosis was detected using flow cytometry and TUNEL method. The opening of mPTP and mitochondrial membrane potential (MMP) were detected by Calcein AM and JC-1 fluorescent probe respectively. The mRNA and protein levels of voltage-dependent anion channel 1 (VDAC1), cyclophilin D (CypD), adenine nucleotide translocator (ANT), cytochrome C (Cyt C), caspase 3, 9 were detected by RT-PCR (real time quantity PCR) and Western blotting respectively. The results demonstrated that mPTP of GC-2 cells was opened after 24 hours of ATR treatment, resulting in decreased MMP and increased apoptosis. Pre-protection with WZYZ-medicated serum and CsA inhibited the opening of mPTP of GC-2 cells induced by ATR associated with increased MMP and decreased apoptosis. Moreover, the results of RT-qPCR and WB suggested that WZYZW-medicated serum could significantly reduce the mRNA and protein levels of VDAC1 and CypD, Caspase-3, 9 and CytC, as well as a increased ratio of Bcl/Bax. However, ANT was not significantly affected. Therefore, these findings indicated that WZYZW inhibited mitochondrial mediated apoptosis by attenuating the opening of mPTP in GC-2 cells. WZYZW-medicated serum inhibited the expressions of VDAC1 and CypD and increased the expression of Bcl-2, which affected the opening of mPTP and exerted protective and anti-apoptotic effects on GC-2 cell induced by ATR.

Mechanism of Dendrobium huoshanense in Treatment of Gastric Ulcer： Based on Network Pharmacology and in Vivo Experiment / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Dendrobium huoshanense in the treatment of gastric ulcer （GU） based on network pharmacology and in vivo experiment. MethodThe active components of D. huoshanense were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and literature， and the targets of the components were screened from TCMSP and SwissTargetPrediction. GU-related genes were retrieved from GeneCards， Online Mendelian Inheritance in Man （OMIM）， and DisGeNET. Thereby， the common targets of the disease and the medicinal were yielded and the protein-protein interaction （PPI） network was constructed， followed by Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. According to the predicted results， hematoxylin-eosin （HE） staining， immunohistochemistry， enzyme-linked immunosorbent assay （ELISA）， Western blot， and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） were used to validate the effects of D. huoshanense on acetic acid-induced GU in rats. ResultA total of 63 active components of D. huoshanense and 37 target genes of D. huoshanense for the treatment of GU were screened out. PPI network analysis yielded several possible core anti-GU targets of D. huoshanense. They influenced the development of GU by acting on signaling pathways such as phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）， hypoxia inducible factor-1 （HIF-1）， tumor necrosis factor （TNF）， and nuclear factor-κB （NF-κB）， and various biological processes. The in vivo experiment showed that D. huoshanense significantly reduced the levels of inflammatory factors such as interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and TNF-α in the serum of model rats （P<0.05， P<0.01）， increased gastric blood flow （GBF） at the ulcer margin， raised the expression of epidermal growth factor （EGF） and epidermal growth factor receptor （EGFR） at the ulcer margin （P<0.01）， significantly down-regulated protein and mRNA expression of PI3K and Akt， and up-regulated protein and mRNA expression of phosphatase and tensin homolog deleted on chromosome ten （PTEN） in the gastric tissues of GU rats （P<0.01）. ConclusionThrough regulating EGFR/PI3K/Akt signaling pathway， D. huoshanense can inhibit tissue inflammation， increase gastric microcirculatory blood flow at the ulcer margin， and promote cell proliferation and repair of damaged gastric mucosa.

Research progress on chemical constituents and pharmacological effects of Glycyrrhizae Radix et Rhizoma and discussion of Q-markers / 中国中药杂志

Glycyrrhizae Radix et Rhizoma, a traditional Chinese herbal medicine, mainly contains triterpenoids, flavonoids, polysaccharides, coumarins and volatile oils with many pharmacological activities such as anti-tumor, anti-bacterial, anti-viral, anti-inflammatory, immune regulatory and anti-fibrotic effects. The widespread applications of Glycyrrhizae Radix et Rhizoma in food, medicine and chemical industries make its demand increase gradually. Therefore, the quality guarantee of the medicinal is of great value. Starting from the elaboration of chemical components and pharmacological effects of Glycyrrhizae Radix et Rhizoma and the introduction to the concept of quality marker(Q-marker), this study analyzed the Q-markers of Glycyrrhizae Radix et Rhizoma from the aspects of plant phylogene-tics, chemical component specificity, traditional efficacy, traditional medicinal properties, absorbed components, different processing methods and so on, which provides reference for quality evaluation, development and utilization of Glycyrrhizae Radix et Rhizoma.

Review of chemical composition, pharmacological effects, and clinical application of Salviae Miltiorrhizae Radix et Rhizoma and prediction of its Q-markers / 中国中药杂志

Salviae Miltiorrhizae Radix et Rhizoma is a Chinese herbal medicine that promotes blood circulation to remove blood stasis, nourishes blood to tranquilize the mind, and cools blood to disperse carbuncles. Salviae Miltiorrhizae Radix et Rhizoma has microcirculation-improving, blood vessel-dilating, atherosclerosis-preventing, anti-inflammatory, anti-tumor, and blood pressure-and blood lipid-lowering activities. As research progresses, the chemical composition, pharmacological effect, and clinical application of Salviae Miltiorrhizae Radix et Rhizoma have attracted much attention. We reviewed the research progress in this field. Based on the concept of quality marker(Q-marker) in traditional Chinese medicine, the Q-markers of Salviae Miltiorrhizae Radix et Rhizoma were predicted and analyzed from the aspects of quality transfer, traceability, ingredient specificity, association between ingredients and pharmacological effects, ingredient predictability, and compounding environment. This review provides a scientific basis for the quality control of Salviae Miltiorrhizae Radix et Rhizoma and its preparations.

Improvement researches on quality standard of Anemarrhenae Rhizoma and its raw processed products / 中国中药杂志

This study is to improve the quality standard and supply the scientific basis for Anemarrhenae Rhizoma and its raw processed products. Steroidal saponin including timosaponin BⅡ, timosaponin AⅢ and flavonoids including neomangiferin and mangiferin were selected as the indicative components. Silica gel G thin layer chromatography(TLC) and polyamide TLC were used to detect the two types of compounds, respectively. The contents of timosaponin BⅡ and timosaponin AⅢ were determined by HPLC-ELSD and the content of neomangiferin, mangiferin and isomangiferin were determined by HPLC-UV. Moisture, total ash and acid insoluble ash were determined according to Chinese Pharmacopoeia(2015 edition). And 80% ethanol was selected as the solvent and the content determination of total extract were determined. The fingerprints of Anemarrhenae Rhizoma and its raw processed products were established by HPLC-UV and HPLC-ELSD. The results showed that the methods of TLC and HPLC have been successfully stablished. There are 2 and 3 peaks which have been identified by HPLC-ELSD and HPLC-UV, respectively. The HPLC fingerprint methods are specific and can be used to identify and quality control for Anemarrhenae Rhizoma and its raw processed products in the mass. Comparing to Chinese Pharmacopoeia(2015 edition), the TLC identification and content determination were revised and the total extract determination and HPLC fingerprints were added in the present study. Our results can be used as the scientific basis of quqlity control for Anemarrhenae Rhizoma and its raw processed products.

Ecology suitability study of Polygonatum cyrtonema / 中国中药杂志

Using the 260 geographical distribution records of Polygonatum cyrtonema in China, combined with 53 environmental factors, the maximum entropy modeling(MaxEnt) was used to study the ecological factors affecting the suitability distribution of P. cyrtonema. The ArcGIS software was used to predict the potential distribution of the population of P. cyrtonema. The dominant factors were chosen by using the Jackknife test and the Receiver Operating Characteristic(ROC) curve was used to evaluate the simulation. The results showed that high value of area under curve(AUC) denoted good results, which significantly differed from random predictions. Based on the evaluation criterion, the accuracies of the predictions of P. cyrtonema potential distribution in the current periods were excellent. The main environmental factors affecting the suitable growth of P. cyrtonema were the monthly precipitation, the wettest monthly precipitation, the annual average temperature range and the precipitation of November, March, February, April, May and October. There are 9 environmental factors in soil type. The potential fitness of P. cyrtonema in China is high, mainly concentra-ted in Hunan, western Hubei, Guangdong, northeastern Guangxi, southeastern Guizhou, Jiangxi, southwestern Anhui, Fujian, Zhejiang, Shaanxi, southwestern Henan and Chongqing. The growth distribution of the potential distribution area of P. cyrtonema was divided, and the zoning map of the growth suitability of P. cyrtonema was formed. Through the comparative analysis of the potential distribution range based on MaxEnt and the distribution range of literature records, the understanding of the distribution range of P. cyrtonema was expanded.

Simultaneous determination and analysis of amino acids and nucleosides in Dendrobium by UHPLC-QTRAP-MS/MS / 中国中药杂志

By using multivariate statistical analysis to evaluate essential quality, and provide scientific basis for their comprehensive utilization, we established an UHPLC-QTRAP-MS/MS method for the fast, precise, efficient determination of 21 kinds of amino acids and 10 kinds of nucleosides in different species of Dendrobium. The analysis was performed on a Waters XBridge Amide column(2.1 mm×100 mm,3.5 μm) with elution by mobile phase of 0.2% formic acid in water-0.2% formic acid in acetonitrile at a flow rate of 0.2 mL·min~(-1) with the column temperature at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring(MRM) mode. The comprehensive evaluation of different species of Dendrobium was carried out by PCA and TOPSIS analysis. All 21 kinds of amino acids and 10 nucleosides showed good linearity among certain concentration range(r>0.999), the RSDs of the stability, precision, and repeatability tests were less than 3.0%. The recovery rate was in the range from 93.31% to 107.5%, and RSD was in the range of 1.1%-3.7%. The comprehensive evaluation index obtained with PCA showed that D. huoshanense was significantly higher than others regarding amino acids and D. officinale has higher nucleosides than other species. The biggest C_i difference of TOPSIS was 68.7%, and comprehensive evaluation showed that D. huoshanense produced the highest comprehensive quality. The method is precise, fast and efficient and can provide reliable basis for further researches and intrinsic quality control of Dendrobium.

Therapeutic Effect and Mechanism of Taohong Siwu Granule on Rats with Dysmenorrhea Due to Cold Coagulation and Blood Stasis / 中国实验方剂学杂志

Objective:To observe the effect of Taohong Siwu granule on dysmenorrhea rats. Method:Forty-eight healthy SD female rats were randomly divided into six groups: control group, model group, Taohong Siwu granule groups (18, 9, 4.5 g·kg-1) and Fuke Qianjin tablet group (0.08 g·kg-1). The rats were given Bugaorer (0.35 mg·kg-1) every day and ice-water bath for 8 minutes for 10 consecutive days. Oxytocin was injected intraperitoneally on the 11th day. The writhing reaction of rats was observed, and the effect of Taohong Siwu granules on hemorheology was measured. The levels of 6-ketoprostacyclin F1α(6-keto-PGF1α), thromboxane B2(TXB2), prostaglandinF2α（PGF2α）, PGE2 of rats were detected by enzyme-linked immunosorbent assay (ELISA). Ca2+ in uterine tissues was scanned by laser confocal microscopy. Result:Each dose of Taohong Siwu granule group reduced the number of writhing, the time of writhing and the incidence of writhing in dysmenorrhea rats, which were significantly lower than those in model group (P<0.05，P<0.01). Compared with the model group, high, medium and low-dose Taohong Siwu granules groups could reduce blood viscosity and plasma viscosity, and Taohong Siwu granules can significantly reduce the content of TXB2, Ca2+，PGF2α, and significantly increase the content of 6-keto-PGF1α，PGE2 (P<0.05，P<0.01). Conclusion:Taohong Siwu granule can significantly alleviate dysmenorrhea symptoms in rats. The mechanism may be correlated with the contents of 6-keto-PGF1α, TXB2 and PGF2α, PGE2 and Ca2+.

Tao-Hong-Si-Wu Decoction promotes angiogenesis after cerebral ischaemia in rats via platelet microparticles / 中国天然药物

Platelet microparticles (PMPs) are membrane particles derived from the platelet membrane that enter into the blood circulation. We sought to explore the therapeutic effects of Tao-Hong-Si-Wu Decoction (THSWD) on angiogenesis in a rat model of cerebral ischaemia-reperfusion (I/R). The protective effect of THSWD on I/R rats was observed morphologically by immunohistochemical expression of VEGF and CD34, along with immunofluorescence results of co-expression of BrdU and vWF. Then, PMPs from different groups of rats were extracted, and cytokine array analysis was used to screen for angiogenesis associated proteins. The results showed that THSWD can promote the expression of VEGF, CD34, BrdU and vWF. Cytokine array analysis revealed the changes in the expression of 29 related angiogenic proteins in the total protein of PMPs, which involved the Notch signalling pathway. Compared with model group, the expression levels of NICD and Hes-1 in the THSWD group were significantly increased. In the context of I/R, the angiogenesis-related proteins of PMPs are different. THSWD may involve the promotion of activation of the Notch signalling pathway to achieve therapeutic effects on cerebral ischaemia.

Cloning and quantitative expression analysis of GMPP gene from Dendrobium huoshanense / 中国中药杂志

In order to understand the function of GDP-mannose pyrophosphorylase(GMPP) function and its regulation in polysaccharide biosynthesis mechanism in Dendrobium. D. huoshanense was used to clone GMPP gene. GMPP gene expression in D. huoshanense,D. officinale and D. moniliforme was also determined by qPCR. The results showed that the length of D. huoshanense GMPP gene c DNA sequence is 1 867 bp,containing 1 245 bp open reading frame(ORF),encoding 415 amino acids. Phylogenetic tree analysis showed that D. huoshanense,D. officinale and D. moniliforme are closely related with GMPP taken into consideration. Bioinformatics analysis demonstrated that GMPP sequence similarity among the three species reached as high as 99%. qPCR results indicated that GMPP genes was highly expressed in stem of D. huoshanense compared with its leaf,flower and root. According to GMPP gene expression profile in D. huoshanense,D. officinale and D. moniliforme grown in Huoshan area,it was clear that GMPP in D. huoshanense showed the highest expression level. Furthermore,our findings of GMPP gene expression profile will facilitate future researches into its polysaccharide biosynthetic mechanism.

Therapeutic effect and mechanism of three kinds of Dendrobium on constipation in rats with spleen Yin deficiency / 中国中药杂志

Books on Chinese herbal medicines have shown that Dendrobium has the effect of nourishing Yin and reinforcing Yin,usually used for constipation induced by spleen Yin deficiency in clinical application. D. huoshanense,as an independent species among many species of Dendrobium,has no experimental studies about its effects on spleen Yin deficiency-type constipation. The purpose of this experiment was to illustrate the therapeutic effect of D. huoshanense on the constipation of spleen Yin deficiency type in rats,investigate its preliminary mechanism,and compare it with the D. officinale and D. nobile contained in the Chinese Pharmacopoeia to clarify its characteristics. The spleen Yin deficiency model was replicated in 70 rats by the composite factor method,and then the model rats were randomly divided into 7 groups: model group,Liuwei Dihuang Pills group( LWDHP),D. huoshanense high( DHS-H),medium( DHS-M),low( DHS-L) dose groups,D. nobile group( DNS),and D. officinale group( DOS),and another 10 rats were used as normal group( Normal). After 7 continuous days of administration,the fecal water content and intestine propulsion rate of each group were detected. HE staining was used to observe the pathological damage of ileum and colon in each group. Immunohistochemistry and Western blot were used to detect aquaporin 3( AQP3) expressions,while the expression levels of the somatostatin( SS) and motilin( MTL) in the ileum of each group were detected by enzyme-linked immunosorbent assay. The results showed that as compared with the model group,the rats in each drug-administered group had increased number of fecal pellets,increased fecal water content,and the increased intestinal propulsion rate( P<0. 01),while the pathological damage of the ileum and colon was significantly reduced; the expression of AQP3 protein was significantly decreased( P<0. 01); the level of MTL was significantly increased and the level of SS was decreased( P<0. 01). All DHS groups showed a good dose-effect relationship,and the same dose treatment effect was equivalent to that of DOS,but it was superior to DNS. Therefore,DHS has a significant therapeutic effect on constipation of spleen Yin deficiency type,and its mechanism may be related to intestinal motility and water-liquid metabolism,with a good therapeutic effect.

Effect of Acori Tatarinowii Rhizoma on Intestinal Absorption of Ginsenosides in Dingzhi Xiaowan / 中国实验方剂学杂志

Objective: To explore the effect of Acori Tatarinowii Rhizoma on intestinal absorption of ginsenosides in Dingzhi Xiaowan,and reveal the mechanism of Acori Tatarinowii Rhizoma acting as " adjuvant drug" in this formula. Method: The contents of ginsenoside Rg1,Re and Rb1 were measured by UPLC-MS/MS and the absorption of three ginsenosides in different intestine segments was investigated by rat single pass intestinal perfusion in situ,including absorption rate constant(Ka) and apparent permeability coefficient(Papp).Everted intestinal sac model was used to investigate the absorption dosage of three ginsenosides affected by volatile oil from Acori Tatarinowii Rhizoma and verapamil[Ver,a P-glycoprotein(P-gp) inhibitor]. Result:Papp values of three ginsenosides were ≤ 0.191×10-3 cm·min-1 in Dingzhi Xiaowan when lack of Acori Tatarinowii Rhizoma.Compared with lack of Acori Tatarinowii Rhizoma in Dingzhi Xiaowan group,the Ka and Papp values of lack of volatile oil from Acori Tatarinowii Rhizoma in Dingzhi Xiaowan group slightly increased without significant difference in the four intestinal segments,but when the prescription had Acori Tatarinowii Rhizoma,the Ka increased by 3.97-8.35 fold and the Papp increased by 3.99-8.49 fold.The results of everted intestinal sac test showed that volatile oil of Acori Tatarinowii Rhizoma could significantly promote the intestinal absorption of ginsenoside Rg1,Re and Rb1,but there was no dose-dependent. Conclusion:Volatile oil of Acori Tatarinowii Rhizoma can promote the intestinal absorption of three ginsenosides in Dingzhi Xiaowan,and the mechanism may be related to the inhibiting function on P-gp.

Anti-inflammatory Effect of Dendrobii Huoshanense Herba / 中国实验方剂学杂志

Objective:To observe the anti-inflammatory effect of Dendrobii Huoshanense Herba. Method:Totally 60 Kunming mice were randomly divided into normal group, model group, high, middle and low-dose Dendrobii Huoshanense Herba groups (DHS-H, DHS-M, DHS-L, 4, 2, 1 g·kg-1) and dexamethasone acetate group (DXMS, 0.01 g·kg-1). The rats were intragastrically administered for 14 days. After the last administration for 1 h, a total of 20 μL of xylene was added to both sides of the right ear center of the mice to establish the ear swelling model. All of the mice were decapitated 1 h later, and the ear swelling inhibition rate of each group were calculated. Totally 36 healthy SD rats were divided into normal model, model group, DHS-H,DHS-M,DHS-L groups (2.8, 1.4, 0.7 g·kg-1) and DXMS acetate group. The rats were intragastrically administered for 7 days. One hour after the last intragastric administration, a foot swelling model was established through subcutaneous injection of 10%fresh egg white in the right hind limb toe of each group. The right hind paw circumference of each rat was measured at 0, 0.5, 1, 2, 3, 4, 5 h, and the paw swelling of the rats was calculated. Totally 60 SD rats were implanted subcutaneously with sterile dry cotton balls in the bilateral groin and grouped as above. All of the rats were intragastrically administered for 14 days since the next day. After the last administration, cotton balls were taken out, and the inhibition rate of granuloma was calculated. And the contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-6 (IL-6), interferon-γ (IFN-γ) in the serum of each group were detected. Result:Compared with the normal group, the ear swelling rate, the foot swelling degree were significantly increased in the model group(PPPPPα, IL-1β, IL-2, IL-6, and IFN-γ in the model group were higher than those in the normal group (Pα, IL-1β, IL-2, IL-6, and IFN-γ in Dendrobii Huoshanense Herba group and positive drug group were decreased significantly (PPConclusion:Dendrobii Huoshanense Herba could effectively inhibit acute and chronic inflammatory reactions.

Simultaneous determination of five saponins in Bupleuri Radix by HPLC-DAD dual wavelength method / 中国中药杂志

Epoxy ether type and isophthalene type saponin are the main saponins of Bupleurum chinense. However,due to the difference of their UV spectrum,there is no quantitative method for simultaneous determination of these two kinds of saponins. In this paper,a dual-wavelength high performance liquid chromatography(HPLC) was developed for simultaneous determination of five saponins in epoxidized ether(saikosaponin a,c,d) and isosorbide type(saikosaponin b1,b2). The mobile phase was eluted with acetonitrile-water(0.1% phosphoric acid) gradient at a column temperature of 30 °C and a flow rate of 1.0 mL·min⁻¹. The detection wavelengths were 208 nm for saikosaponins a,c, and d, and 254 nm for saikosaponins b₁ and b₂. The results showed that the separation of five kinds of saikosaponin was good, with the linear range of 9.70-1 935.00(=0.999 4),8.20-1 380.00(=0.999 3),6.90-1 640.00(=0.999 0),5.25-630.00(=0.999 4), and 5.15-618.00 mg·L⁻¹(=0.999 5), respectively. The average recoveries were 97.70%-100.2% and the RSD was less than 3%(=6). The method is simple,rapid and reproducible. It can be used for the determination of five kinds of saikosaponins in B. chinense.

Comparison of different extraction methods of exosome from macrophages / 中国药理学通报

Aim To establish the methods for exosomes isolation from the cell culture medium based on the principle of polyethy-lene glycol (PEG) precipitation combined with ultra-high speed centrifugation. Methods Exosomes were extracted by different centrifugation, PEG6000 precipitation combined with ultra-high speed centrifugation and EXO Quick kit,respectively. The mor-phology and particle size of exosomes were analyzed by transmis-sion electron microscopy,NanoSight ZS and Nano-particle track-er(NTA). Western blot assay was used to identify if the extrac-tion were exosomes. Results According to PEG6000 precipita-tion combined with ultra-high speed centrifugation method, the exosomes were successfully isolated from the culture medium. The morphology and particle size of exosomes,the exosomes iso-lated from PEG6000 precipitation combined with ultra-high speed centrifugation were equal to those of the ultra-high speed centrifugation but smaller to EXO Quick kit. What's more, the results of transmission electron microscopy showed that the three methods had typical discoid vesicles. Conclusion The im-provement of the exosome isolation from cell culture medium is convenient,easy and cheap.

Next generation sequencing and transcriptome analysis of root bark from Paeonia suffruticosa cv. Feng Dan / 中国中药杂志

Moutan Cortex is an important traditional Chinese medicine, "Fengdan Pi" was known as Dao-di herbs from the root bark of Paeonia suffruticosa cv. Feng Dan for its extracted various active components. However, the genetic basis for their activity is virtually unknown. The transcriptome of the root bark from "Fengdan" was sequenced using the Illumina HiSeq 4000 sequencing platform. The clean reads were then de novo assembled into 72 997 unigenes. Among them, the number of unigenes which could been annotated by dataset Nr and GO was 41 139 and 34 592. The 20 016 unigenes could been annotated by KEGG dataset, which were involved in 5 major categories, 34 middle categories, and 352 metabolism pathways. The number of unigenes which were mapped to the phenylpropanoid biosynthesis pathway, terpenoid backbone biosynthesis pathway, terpenoid biosynthesis pathway, alkaloid biosynthesis pathway, and flavonoid biosynthesis pathway was 214, 104, 152, 55 and 36 respectively, suggesting that they are involves in these pathways of pharmaceutically important. Furthermore, there also showed remarkable differences in groups which enrichment ratio of the different expressed gene compared. In addition, a total of 9 939 SSRs were identified from the sequence of 72 997 unigenes. This study not only provides many valuable basal data which was important gene in the synthesis pathway of secondary metabolites with gene searching, but also has important significance to find molecular marker in germplasm for breeding and improvement.

Formation and definition of Dao-di herbs："Wan medicines" / 中国中药杂志

As the saying goes, bencao is growing in specific areas. Anhui province is located in the middle and lower reaches of the Yangtze River Plain. Its north-south direction is longer than the east-west and Anhui is divided into three regions by the Yangtze River and Huaihe. It is important for the land to have plentiful traditional Chinese medicine resources, and it exists profound cultural heritage of Chinese medicine and to cluster the Dao-di herbs with a long history. The vicissitude of historical geography of Anhui province was combed, the historical basis of Anhui short as "Wan" was hackles, the prosperity and decline of Dao-di herbs in Anhui territory was sorted out in this paper. This article holds that the history of Anhui established as a province started in the Qing Dynasty, but the history of Dao-di herbs in Anhui territory has always been the same strain, Bencao can be test clearly and production areas is stable. Despite Anhui Dao-di herbs have changed in the different historical periods, its core varieties and essential elements has been inherited to today. Generally speaking, "Wan medicines" are not only collectively called the current territory of Anhui Dao-di herbs, but also include the historical period of Anhui famous Dao-di herbs.

Variety and distribution of Anhui native medicinal materials in ancient Chinese materia medica / 中国中药杂志

Anhui is located in the middle and lower reaches of the Yangtze River Plain, its across warm temperate zone and subtropics. The mountain and water next to each other, which leads to Chinese medicine resources ranked first in East China. The utilization of traditional Chinese medicine resources in Anhui has a long history, which could date back to the publishing time of Ming Yi Bie Lu (Appendant Records of Famous Physicians). And the kinds of traditional Chinese medicine in Song Dynasty ups to 80. There are also some differences in the distribution of various geographical units in terms of the types: Jianghuai hilly region's ups to 64, 25 in Wannan mountainous area, the species in Dabie Mountains and Huaibei plain are 16 and 14 respectively. In addition, the Jianghuai hilly region's and Wannan mountainous area have a long history among of them, which have been reached a peak in the Song Dynasty. The history of native medicinal materials in Anhui recorded in different periods, though combing herbal books. And the results showed that the vast majority of varieties in ancient are the same as modern ones, which provide the historical basis for the rich bulk medicinal materials in Anhui. The distinctions in natural and social environment of different geographical units have effects on the history of the usage of Chinese medicine resources in respective regions. Thus, the variety and distribution of native medicinal materials in Anhui among the Bencao works of different period provides herbalism basis for the protection and utilization of Chinese medicine resources currently.

Geographical distribution of Chinese herbalists：causes of four distribution centers about Huizhou, Shanghai, Hangzhou, Suzhou / 中国中药杂志

Based on the "Zhong Guo Ben Cao Yao Ji Kao", Chinese herbalists in past dynasties were counted and analyzed by their living period,numbers and native places. Combined with GIS, the geographical distribution and the formation causes of the four distribution centers of herbalists in past dynasties were discussed. The results showed that, there was a greater difference between the numbers of herbalists in different periods of time, which achieved to the peak in the Ming and Qing Dynasties. In addition, the distribution of herbalists in past dynasties characterized east more and west less, forming the distribution areas centered by Huizhou, Shanghai, Hangzhou and Suzhou. Besides, the geographical distributions of Chinese herbalists showed an obvious southward trend since the Song Dynasty.