Specific PCR for Identification of Astragalus membranaceus var. mongholicus Seeds， A. membranaceus Seeds， and Adulterants / 中国实验方剂学杂志

ObjectiveTo establish a method based on specific polymerase chain reaction （PCR） that can accurately and rapidly identify Astragalus membranaceus var. mongholicus （AMM） seeds and A. membranaceus （AM） seeds. MethodThe Chloroplast Genome Information Resource （CGIR） and IdenDSS were used to obtain the characteristic DNA fragments of AMM and AM， and the specific single nucleotide polymorphism （SNP） sites of AMM and AM were screened out， on the basis of which the specific primers MG-F/MG-R of AMM and MJ-F/MJ-R of AM were designed. The specific PCR method for identifying AMM and AM was established and optimized， and the specificity and applicability of the method were investigated. ResultThe specific PCR conditions for the identification of AMM were primers MG-F/MG-R， annealing at 62 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the specific band appeared at about 220 bp， with no band for the seeds of AM or adulterants. The specific PCR conditions for identifying the AM were primers MJ-F/MJ-R， annealing at 58 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the band appeared at about 150 bp， with no band of AMM or adulterants. ConclusionThe specific PCR method established in this study can accurately and quickly identify the seeds of AMM and AM， which provides a basis for the classification and accurate identification of Astragalus seeds and adulterants.

Progress in Mechanism of Astragalus membranaceus and Its Chemical Constituents on Multiple Sclerosis / 中国结合医学杂志

The primary chemical components of Astragalus membranaceus include polysaccharides, saponins, flavonoids, and amino acids. Recent studies have shown that Astragalus membranaceus has multiple functions, including improving immune function and exerting antioxidative, anti-radiation, anti-tumor, antibacterial, antiviral, and hormone-like effects. Astragalus membranaceus and its extracts are widely used in clinical practice because they have obvious therapeutic effects against various autoimmune diseases and relatively less adverse reaction. Multiple sclerosis (MS) is an autoimmune disease of central nervous system (CNS), which mainly caused by immune disorder that leads to inflammatory demyelination, inflammatory cell infiltration, and axonal degeneration in the CNS. In this review, the authors analyzed the clinical manifestations of MS and experimental autoimmune encephalomyelitis (EAE) and focused on the efficacy of Astragalus membranaceus and its chemical components in the treatment of MS/EAE.

Optimization of extraction technology for couplet medicinals of Astragalus membranaceus-Puerariae lobata by multi-index comprehensive scoring method combined with orthogonal experiment / 中国药房

OBJECTIVE To optimize extraction technology of couplet medicinals of Astragalus membranaceus-Puerariae lobatae. METHODS With contents of puerarin，daidzin，calycosin-7-O-β-D-glucopyranoside，daidzein，calycosin and formononetin and the yield of dry extract as index，the analytic hierarchy method was used to determine the weight coefficient of each index and calculate the comprehensive score. The effects of solid-liquid ratio， extraction times and extraction time on the comprehensive score were investigated by single factor test. The level of each factor was determined. By multi-index comprehensive scoring method， using comprehensive scores of above 7 indexes as indexes，the extraction technology of couplet medicinals of A. membranaceus-P. lobata was optimized by orthogonal experiment，and the validation tests were conducted. RESULTS The weight coefficient for the contents of puerarin，daidzin，calycosin-7-O-β-D-glucopyranoside，daidzein，calycosin and formononetin and the yield of dry extract were respectively 0.304 7，0.065 2，0.185 8，0.185 8，0.107 8，0.107 8 and 0.042 7. The optimal extraction technology was determined as follows： solid-liquid of 1∶8（g/mL），extracting 3 times and for 1 h each time. RSD of each evaluation index in the validation test results was lower than 3.00% （n＝3）. CONCLUSIONS The optimized extraction technology for A. membranaceus-P. lobata is stable and feasible.

Inhibitory effect of astragalus membranaceus injection on apoptosis and autophagy induced by enterovirus 71 / 中国生物制品学杂志

@#Objective To investigate the effect of astragalus membranaceus（AM）injection on apoptosis and autophagy of human gastric epithelial cell line（GES⁃1）induced by enterovirus 71（EV71）. Methods GES⁃1 cells were cultured in vitro and divided into infected group（EV71 infected at a MOI of 3 and control group（no virus infected）. The morpho⁃logical changes of EV71 infected cells were observed by inverted microscope. The level of VP1 in GES⁃1 cells infected with EV71 was detected by Western blot；CCK⁃8 assay was used to detect the viability of GES⁃1 cells infected with EV71；Nuclear staining with DAPI was used to observe the morphological changes of nuclear apoptosis infected with EV71. GES⁃1 cells were divided into control group（without virus infection），infection group and AM intervention group with final concentration of 1，2. 5，5 and 10 μg／mL，respectively. Western blot was used to detect the effect of AM intervention on the expression of apoptosis⁃related proteins Caspase⁃3，PARP and autophagy⁃related proteins LC3 and P62 in GES⁃1 cells infected withEV71. CCK⁃8 method was used to detect the effect of AM intervention on the viability of GES⁃1 cells infected with EV71. Results GES⁃1 cells were round，shrunken with nuclear pyknosis and uneven size；VP1 level increased（t = 41. 56，P < 0. 01），cell viability decreased（t = 19. 07，P < 0. 01），Caspase⁃3 and PARP proteins were cut off（t = 35. 29 and 3. 648， P < 0. 01 and 0. 021 8，respectively），LC3Ⅱ／LC3Ⅰ ratio increased（t = 10. 16，P = 0. 000 5）and P62 protein was degraded（t = 68. 68，P < 0. 01）；AM inhibited the degradation of Caspase⁃3，PARP and P62 proteins induced by EV71 （t = 52. 66，59. 60 and 40. 22，respectively，each P < 0. 01）and increased the ratio of LC3Ⅱ／LC3Ⅰ（t = 5. 521，P = 0. 005 3），andreducedtheinhibitoryeffectofEV71ontheviabilityofGES⁃1cells（t =4. 420，P =0. 0115）. Conclusion EV71 infection induced apoptosis of GES⁃ 1 cells and AM intervention inhibited EV71 induced apoptosis by inhibiting EV71 induced autophagy.

Chemical composition analysis and value evaluation of stems and leaves of Astragalus membranaceus var. mongholicus / 中国中药杂志

This study aimed to provide data support for resource utilization of the stems and leaves of Astragalus membranaceus var. mongholicus(SLAM) by analyzing and evaluating the chemical constituents. The crude protein, crude fiber, and soluble saccharide of SLAM were analyzed by Kjeldahl method, filtration method, and UV-Vis spectrophotometry, respectively. The nucleosides, amino acids, flavonoids, and saponins of SLAM were analyzed by ultraperformance liquid chromatography-triple quadrupole mass spectrometry(UPLC-TQ-MS). Combined with principal component analysis(PCA), the quality difference of resource components of SLAM was comprehensively evaluated. The results showed that the average content of crude protein, crude fiber, total polysaccharide, and redu-cing sugar in SLAM was 5.11%, 30.33%, 11.03 mg·g~(-1), and 31.90 mg·g~(-1), respectively. Six nucleosides, 15 amino acids, 22 flavonoids, and one saponin were detected, with an average content of 1.49 mg·g~(-1), 6.00 mg·g~(-1), 1.86 mg·g~(-1), and 35.67 μg·g~(-1), respectively. The content of various types of chemical components in SLAM differed greatly in different harvesting periods and growing years. The results of PCA showed that the quality of SLAM produced in Ningxia was superior. The results can provide references for the utilization of SLAM.

Protective effect and mechanism of Astragalus membranaceus and Angelica sinensis compatibility against triptolide-induced hepatotoxicity by regulating Keap1/Nrf2/PGC-1α / 中国中药杂志

This paper aims to investigate the protective effect and mechanism of Astragalus membranaceus and Angelica sinensis before and after compatibility against triptolide(TP)-induced hepatotoxicity. The experiment was divided into a blank group, model group, Astragalus membranaceus group, Angelica sinensis group, and compatibility groups with Astragalus membranaceus/Angelica sinensis ratio of 1∶1, 2∶1, and 5∶1. TP-induced hepatotoxicity model was established, and corresponding drug intervention was carried out. The levels of alanine transaminase(ALT), aspartate transaminase(AST), and alkaline phosphatase(ALP) in serum were detected. Pathological injuries of livers were detected by hematoxylin-eosin(HE) staining. The levels of malondialdehyde(MDA), superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), and reduced glutathione(GSH) in the liver were measured. Wes-tern blot method was used to detect the expression of nuclear factor erythroid 2-related factor 2(Nrf2), Kelch-like ECH-associated protein 1(Keap1), peroxisome proliferator-activated receptor gamma, coactivator-1 alpha(PGC-1α), heme oxygenase-1(HO-1), and NAD(P)H quinone dehydrogenase 1(NQO1) in livers. Immunofluorescence was used to detect the expression of Nrf2 and PGC-1α in livers. The results indicated that Astragalus membranaceus/Angelica sinensis ratio of 2∶1 and 5∶1 could significantly reduce the levels of serum AST, ALT, and ALP, improve the pathological damage of liver tissue, increase the levels of GSH and GSH-Px, and reduce the content of MDA in liver tissue. Astragalus membranaceus/Angelica sinensis ratio of 1∶1 and 2∶1 could significantly improve the level of SOD. Astragalus membranaceus and Angelica sinensis before and after compatibility significantly increased the protein expression of HO-1 and NQO1, improved the protein expression of Nrf2 and PGC-1α, and decreased the protein expression of Keap1 in liver tissue. The above results confirmed that the compatibility of Astragalus membranaceus and Angelica sinensis had antioxidant effects by re-gulating Keap1/Nrf2/PGC-1α, and the Astragalus membranaceus/Angelica sinensis ratio of 2∶1 and 5∶1 had stronger antioxidant effect and significantly reduced TP-induced hepatoto-xicity.

Research progress of effective components of Astragalus on prevention and treatment of organ fibrosis / 药学学报

Astragalus, which was first documented in Shennong Bencao Jing, is the dried root of Astragalus membranaceus (Fisch.) Bge. or Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. The active ingredients astragalus membranaceus saponins (AMS), astragalus polysaccharides (APS) and astragalus flavonoids (AFS) have pharmacological effects such as anti-tumor properties, lowering blood sugar, regulating lipid metabolism, cardiovascular protection, anti-oxidation, bone protection, anti-fibrosis, etc. Fibrosis affects almost all organs, particularly vital organs such as the lungs, liver, heart and kidneys. The primary pathological changes of fibrosis involve abnormal increase of myofibroblasts and excessive deposition of extracellular matrix (ECM) components, which lead to the formation of scar tissue, ultimately resulting in fibrosis and even functional loss or failure of organs, which seriously threatens human health and life. Recent, studies have shown that Astragalus membranaceus has a good therapetuic effect on organ fibrosis. This article reviews the current advances of Astragalus in the prevention and treatment of fibrosis of lungs, liver, heart, kidneys and other important organs.

Analysis of Fusarium oxysporum infection on changes of Astragalus metabolism by metabolomic approach / 药学学报

italic>Astragalus is a commonly used Chinese medicinal material in traditional Chinese medicine (TCM), and with the increase of planting area in recent years, the damage of Astragalus root rot has worsened year by year, which seriously affecting its quality and yield. Fusarium oxysporum is one of the main pathogens causing root rot in astragalus. In this study, UPLC-Q-TOF-MS based metabolomic approach combined with multivariate statistical analysis were used to analyze the metabolite changes of Astragalus in response to F. oxysporum infection. The results showed that 62 metabolites in the Astragalus had significant changes after inoculation of F. oxysporum. Polar metabolites included 40 flavonoids, 8 saponins, 2 nucleosides, 1 vitamin, 1 organic acid, 1 amino acid; while lipid metabolites included 3 fatty acids, 1 diradylglycerols, 2 lysophosphatidylcholine, 1 lysophosphatidylglycerol, 1 phosphatidylinositol, 1 sterol lipid. Among these differential metabolites, the relative content of flavonoids, vitamin B2, tryptophan and salicylic acid were increased, while the relative content of saponins were decreased. Correlation analysis showed that the flavonoids were positively correlated with each other, and positively correlated with most lipids, but negatively correlated with most saponins. In addition, studies have shown that F. oxysporum infection is not an influencing factor for the generation of malonyl substitution of flavonoid. This study elucidates the effect of F. oxysporum infection on Astragalus from the perspective of plant metabolism, which provides a basis for exploring the interaction mechanism between the Astragalus and F. oxysporum and further promoting molecular breeding.

Targeted metabolomic analysis of phenylpropanoid metabolites in Astragalus membranaceus var. mongholicus in response to Fusarium solani infection / 药学学报

Root rot severely restricts the sustainable development of Astragalus membranaceus var. mongholicus (AMM) industry. Resistance breeding is an economical and environmentally safe way to manage the disease and its key lies in the obtaining of resistance indicators. This study aimed to quickly and accurately screen the resistance-related (RR) metabolites so as to provide reference for the screening of indicators of AMM breeding for resistance. LC-MS-based targeted metabolomics and real-time quantitative PCR technology were employed, in combination with multivariate statistical analysis, in analyzing the dynamic changes of phenylpropanoid metabolites in AMM in response to root rot pathogen Fusarium solani (FS) infection and identifying the differential metabolites. The LC-MS method established showed high sensitivity; each metabolite had a good linear relationship (R2 ≥ 0.968 9) in the corresponding linear range of the respective standard curve; the recoveries and the relative standard deviations (RSDs) (n = 6) ranged from 70% to 107% and from 1.2% to 9.9%, respectively. Obvious disturbances were observed in the changes of the targeted metabolites in AMM infected by FS. These metabolites, compared with the mock-inoculated (CK) group, showed different up or down regulation with time series. Calycosin-7-O-β-D-glucoside, ononin, calycosin and formononetin were identified as differential metabolites, and they all belong to flavonoids. The first three compounds were significantly negatively correlated (r ≤ -0.97, P < 0.05) with the content of FS in the root of AMM. As potential RR metabolites, they are helpful in obtaining promising resistance indicators for AMM against FS infection.

Functional characterization of a cycloartenol synthase and four glycosyltransferases in the biosynthesis of cycloastragenol-type astragalosides from Astragalus membranaceus

Astragalosides are the main active constituents of traditional Chinese medicine Huang-Qi, of which cycloastragenol-type glycosides are the most typical and major bioactive compounds. This kind of compounds exhibit various biological functions including cardiovascular protective, neuroprotective, etc. Owing to the limitations of natural sources and the difficulties encountered in chemical synthesis, re-engineering of biosynthetic machinery will offer an alternative and promising approach to producing astragalosides. However, the biosynthetic pathway for astragalosides remains elusive due to their complex structures and numerous reaction types and steps. Herein, guided by transcriptome and phylogenetic analyses, a cycloartenol synthase and four glycosyltransferases catalyzing the committed steps in the biosynthesis of such bioactive astragalosides were functionally characterized from Astragalus membranaceus. AmCAS1, the first reported cycloartenol synthase from Astragalus genus, is capable of catalyzing the formation of cycloartenol; AmUGT15, AmUGT14, AmUGT13, and AmUGT7 are four glycosyltransferases biochemically characterized to catalyze 3-O-xylosylation, 3-O-glucosylation, 25-O-glucosylation/O-xylosylation and 2'-O-glucosylation of cycloastragenol glycosides, respectively. These findings not only clarified the crucial enzymes for the biosynthesis and the molecular basis for the structural diversity of astragalosides in Astragalus plants, also paved the way for further completely deciphering the biosynthetic pathway and constructing an artificial pathway for their efficient production.

A new diphenyl butanedione from Astragalus membranaceus of northern Shanxi / 药学学报

Seven compounds were isolated from Astragalus membranaceus of northern shaanxi by silica gel and Sephadex LH-20 column chromatographies. Their chemical structures were identified on the basis of their physical and chemical properties. These compounds were elucidated as astragaloside IV (1), formononetin (2), calycosin (3), 1-(4-hydroxyphenyl)-4-(2,4-hydroxyphenyl)-2-hydroxy-1,4-but anedione (4), (E)-4-methylcinnamic acid (5), quercetin (6), and uridine (7). Compound 4 is a new compound and compound 5 was isolated from the plants of Astragalus Linn. for the first time. The results of in vitro antitumor activity assay showed that compound 4 could inhibit the proliferation of A549 with IC50 values of 11.41 μmol·L-1.

Study on the mitigation effects and mechanism of Astragalus membranaceus polysaccharides on monocrotaline- induced pulmonary hypertension in rats / 中国药房

OBJECTIVE To study the mitigation effect and its possible mechanism of Astragalus membranaceus polysaccharide on the pulmonary hypertension induced by monocrotaline in rats. METHODS One hundred SD male rats were randomly divided into normal control group ，monocrotaline group ，A. membranaceus polysaccharide low-dose and high-dose groups. In addition to the normal control group, rats in other groups were injected with monocrotaline by single intraperitoneal injection of 60 mg/kg. On days 2 to 28 after administration ，rats in the A. membranaceus polysaccharide low-dose and high-dose groups were intraperitoneally injected with A. membranaceus polysaccharide of 200 mg/kg and 400 mg/kg，respectively，once a day. There were 25 rats in each group，and 15 rats were taken for index detection. The mean pulmonary artery pressure （mPAP）and right heart hypertrophy index （RVHI）were detected ，and morphology changes of pulmonary artery and cardiomyocytes were monitored . mRNA and protein expression of IL- 17 in their lung tissues were detected. RESULTS Compared with normal control group ，mPAP and RVHI of monocrotaline group and A. membranaceus polysaccharide groups were increased significantly （P＜0.01）；mRNA and protein expression of IL- 17 in lung tissues were significantly increased （P＜0.01），and there were obvious pathological changes in pulmonary artery and cardiomyocytes. Compared with monocrotaline group ，mPAP and RVHI were significantly decreased in A. membranaceus polysaccharide groups （P＜0.01），while mRNA and protein expression of IL- 17 in lung tissue were decreased significantly （P＜0.01）；pathological changes in pulmonary artery and cardiomyocytes were improved. Compared with A. membranaceus polysaccharide low-dose group ，above indexes and pathological changes were improved significantly in high-dose group. CONCLUSIONS A. membranaceus polysaccharide can reduce monocrotaline-induced pulmonary hypertension ，improve pulmonary artery structure and myocardial remodeling in rats ， the mechanism of which is presumably related to the down-regulation of IL- 17 expression in lung tissue of rats.

Effect and molecular mechanism research of Astragalus membranaceus on inhibiting intestinal absorption of six alkaloids of Aconitum carmichaelii in spleen deficiency rats / 中草药·英文版

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.

Simultaneous quantitative analyses of six components in Astragalus membranaceus based on HPLC-CAD and quantitative analysis of multi-components with a single-marker / 药学学报

A quantitative analytical method based on HPLC coupled with the charged aerosol detector (CAD) for quantitative analysis of multi-components with a single marker (QAMS) was established for simultaneous determinations of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan in Astragalus membranaceus. The separation was performed on an Agilent SB-C18 (150 mm×4.6 mm, 3.5 μm), with gradient elution using the mobile phase consisting of 0.05% formic acid solution and 0.05% formic acid acetonitrile at the flow rate of 1.0 mL·min-1. The column temperature was 35 ℃, and the injection volume was 20 μL. For CAD, the drift tube temperature was at 50 ℃. The contents of six components in A. membranaceus were determined by both external standard method (ESM) and QAMS, and then were compared. The results showed that chromatographic peaks were separated well and the linear ranges of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were 0.113-2.250 mg·mL-1, 0.012-0.240 mg·mL-1, 0.004-0.080 mg·mL-1, 0.065-1.300 mg·mL-1, 0.005-0.100 mg·mL-1 and 0.007-0.150 mg·mL-1, respectively. The content ranges of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were 0.306-0.922 mg·g-1, 0.053-0.183 mg·g-1, 0.015-0.092 mg·g-1, 0.069-0.823 mg·g-1, 0-0.098 mg·g-1 and 0.020-0.107 mg·g-1 in 20 batches of A. membranaceus, respectively. Using astragaloside Ⅱ as an internal reference, the relative correlation factors of astragaloside Ⅰ, astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, formononetin, and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were calculated as 0.561, 0.835, 0.299, 0.796, and 0.799, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method, and there was no significant difference in assay results between the two methods. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the contents such as astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan, and it can be used for quality control of A. membranaceus.

Metabonomics Study on the Effects of Secondary Components of Astragalus membranaceus Injection on Leuko- penia Model Mice / 中国药房

OBJECTIVE：To study metabonomics of secondary components of Astragalus membranaceus injection on leukopenia model mice. METHODS ：The mice were divided into normal group ，model group ，A. membranaceus injection group （0.004 mL/g），secondary components low-dose ，medium-dose and high-dose groups （0.004，0.008，0.016 mL/g），with 7 mice in each group. Except for normal group ，other groups were given cyclophosphamide （80 mg/kg） intraperitoneally to induce leukopenia model. After modeling ，administration groups were given relevant medicine intraperitoneally ，and normal group and model group were given constant volume of sterile water for injection intraperitoneally ，once a day ，for consecutive 7 days. During the experiment ，the changes of body weight of mice were measured every 2 days. After the last administration ，the blood routine indexes [white blood cell （WBC），neutrophil（NE），lymphocyte（LY）and monocyte （MO）counts] of mice were detected by animal blood analyzer. UPLC-Q Exactive Orbitrap-HRMS combined with multivariate statistical analysis were used to analyze the metabonomics of mice serum. RESULTS ：Compared with model group ，body weight of mice in the secondary component low-dose group increased significantly on the 4th and 8th day of administration （P＜0.05），and the counts of WBC ，NE，LY and MO in serum of mice in secondary component groups increased significantly （P＜0.05 or P＜0.01）. Metabonomic analysis showed that the secondary components of A. membranaceus injection could significantly regulate the contents of 8 endogenous metabolites in serum， including spermidine ， uric acid ， citric acid ， nicotinamide， eicosapentaenoic acid ， linoleic acid ， erucamide and sphingosine-1-phosphate. Their effects involved linoleic acid metabolism pathway ，nicotinic acid and nicotinamide metabolism pathway and tricarboxylic acid cycle pathway. CONCLUSIONS ：The secondary components of A. membranaceus injection possess the effect of increasing white blood cells in leukopenia model mice ，the mechanism of which may be related to intervention of linoleic acid metabolism pathway ， nicotinic acid and nicotinamide metabolism pathway ，and tricarboxylic acid cycle pathway.

Polysaccharide from Astragalus membranaceus promotes the activation of human peripheral blood and mouse spleen dendritic cells / 中国天然药物

Astragalus membranaceus (A. membranaceus) is a widely used traditional herb in China and Korea. A. membranaceus polysaccharides (AMP), which make up a major part of the root extract, have been shown to modulate immune modulations, especially activation of bone marrow-derived dendritic cells (BMDCs) and T cells. However, the immune stimulatory effect of AMP in the mouse in vivo and human peripheral blood DCs (PBDCs) has not been well investigated. In this study, we found that intravenous (i.v.) injection of AMP in C57BL/6 mice induced remarkable elevations in co-stimulatory and MHC class I and II molecule levels in the splenic DCs and its subsets. The stimulatory effect of DCs by AMP was elevated 6 h after treatment, which rapidly decreased 18 h after injection. Furthermore, AMP promoted intracellular production of pro-inflammatory cytokines in spleen DC subsets, which contributed elevation of serum cytokine levels. Finally, the AMP promoted PBDC activation. Thus, these results demonstrate that AMP can be used as an immune stimulatory molecules in human and mouse.

Effect of EBR on Seed Germination， Seedling Growth and Key Enzyme Activities for Flavonoid Biosynthesis in Astragalus membranaceus var. mongholicus Under PEG-induced Drought Stress / 中国实验方剂学杂志

Objective:To study the effects of 24-epibrassinolide （EBR） at different concentrations on seed germination， seedling growth and key enzyme activities for flavonoid biosynthesis in <italic>Astragalus membranaceus </italic>var.<italic> mongholicus</italic> under polyethylene glycol （PEG）-induced drought stress， in order to provide theoretical reference for standardizing the drought-resistant cultivation techniques of <italic>A. membranaceus </italic>var.<italic> mongholicus</italic>. Method:The seeds of <italic>A. membranaceus </italic>var.<italic> mongholicus </italic>were soaked in EBR solution at different concentrations（0.001， 0.01， 0.1， 1， 10 μmol·L^-1） followed by foliar spraying to explore their effects on seed germination， seedling growth， photosynthesis， chlorophyll and malondialdehyde （MDA） contents， and key enzyme activities for flavonoid biosynthesis under drought stress induced by 20% polyethylene glycol （PEG）-6000. Result:Compared with the control（CK）group， PEG-induced drought stress led to an obvious decrease in germination potential， germination rate， germination index， vigor index， relative germination rate， plant height， root length， aboveground and root dry weight， net photosynthetic rate （<italic>Pn</italic>）， stomata conductance （<italic>Gs</italic>）<italic>， </italic>transpiration rate （<italic>Tr</italic>）， chlorophyll contents and chlorophyll a/b ratio， while a significant increase in intercellular carbon dioxide concentration （<italic>Gi</italic>）， MDA， flavonoids contents， and key enzyme activities for flavonoid biosynthesis such as phenylalanine ammonia lyase （PAL）. The treatment with exogenous EBR solution at the suitable concentration significantly enhanced the adaptation of <italic>A. membranaceus </italic>var.<italic> mongholicus </italic>seeds and seedlings to PEG-induced drought stress， manifested as significantly elevated germination potential， germination rate， germination index， vigor index， relative germination rate， plant height， root length， aboveground and root dry weight， <italic>Pn</italic>， <italic>Gs， Tr</italic>， chlorophyll a/b ratio， chlorophyll and flavonoids contents and key enzyme activities for flavonoid biosynthesis like PAL while lowered <italic>Ci</italic> and MDA contents. The optimal concentration of EBR solution was 0.1 μmol·L^-1. Conclusion:Exogenous EBR solution at the suitable concentration ameliorates the inhibitory effect of 20% PEG stress against seed germination and seedling growth of <italic>A. membranaceus </italic>var.<italic> mongholicus</italic>， reduces the oxidative damage in leaves， and improves the stress resistance to a certain extent by up-regulating the key enzyme activities and promoting flavonoid synthesis.

The anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription based on network pharmacology / 中国药理学通报

Aim To explore the anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription based on network pharmacology. Methods The active components and targets of Shuanghuang Qizhen Keliu Prescription were searched and selected through the TCM-SP database. Related targets of tumor were screened and retrieved by OMIM. The key anti-tumor targets of Shuanghuang Qizhen Keliu Prescription were obtained by using Venn diagram analysis. The key target PPI network was constructed and the key targets were obtained by Cytoscape. Metascape platform was used for GO and KEGG enrichment analysis of key targets. Results There were many active components such as quercetin, kaempferol, and beta-sitosterol in Shuanghuang Qizhen Keliu Prescription, and their targets might be TP53, AKT1, JUN, MAPK1, TNF, RELA, IL-6 and so on. The anti-tumour mechanism might be proliferation inhibition, migration inhibition and apoptosis promotion. Conclusions This study preliminarily discusses the potential anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription, providing ideas and basis for subsequent experimental research.

Ater-soluble Protein Profiles of Astragalus membranaceus var. mongholicus and A. membranaceus / 中国实验方剂学杂志

Objective::Astragalus membranaceus var. mongholicus and A. membranaceus are medicinal Astragalus, which are closely related and similar in composition, but with unclear medicinal value. Water-soluble protein profiles for A. membranaceus var. mongholicus and A. membranaceus were established to explore the differences between the two kinds of Astragalus Radix. Method::The water-soluble protein components were obtained through water ultrasonic extraction and acetone precipitation. After digested with trypsin, the obtained peptides were analyzed by nano ESI-LC-MS/MS method. Proteome Discovery 1.4 software was used to identify the proteins by comparing with the legume protein database, and the different expression water-soluble proteins were analyzed by the label-free quantitative software SIEVE. Finally, relevant information for common expression proteins, including classification, molecular function, involved biological process and signaling pathway, were analyzed by bioinformatics. Result::There were 920 and 717 specific proteins identified for A. membranaceus var. mongholicus and A. membranaceus, respectively. Totally 472 proteins were found to be co-expressed, in which 21 were differentially expressed, such as PR-10 protein, NDK-1 protein, glutelin A2, and phospholipase D. There were 14 highly expressed proteins in A. membranaceus var. mongholicus and 7 highly expressed proteins in A. membranaceus. Conclusion::There are significant differences in water-soluble protein profiles for two kinds of Astragalus Radix. Specific proteins, differentially expressed proteins and common expressed proteins can provide references for the identification of A. membranaceus var. mongholicus and A. membranaceus. It also can be used to define pharmacological mechanisms and search for drug action targets.

Effect of Transplanting Densities on Growth, Development, Yield and Quality of Astragalus membranaceus var. mongholicus / 中国实验方剂学杂志

Objective::Astragali Radix is an important medicinal and edible herb. To achieve standardized cultivation of Astragali Radix and improve cultivation results, Astragalus membranaceus var. mongholicus was cultivated with a row spacing of 30 cm and different plant spacing (8, 10, 12, 14, 16 cm) in a test base of Longxi County, Gansu Province. Method::The growth and development dynamics of green strains and the yield and quality of medicinal materials were measured, and the comprehensive evaluation of membership function was used to determine the rational transplanting density and explore the standard production technology of A. membranaceus var. mongholicus. Result::The transplanting topping had a significant effect on the growth and development of A. membranaceus var. mongholicus under the condition of the row spacing of 30 cm. With the increase of topping, the biomass of astragalus on the ground decreased, and the ratio of root to shoot increased, but the yield of medicinal material per unit area decreased. The appearance traits were improved as the transplant density decreased. Under the row spacing of 14 cm, the content of astragaloside and calycosin-7-glucoside were the highest. Under the row spacing of 8-16 cm, the content of root extract of Astragali Radix was better than the Chinese Pharmacopoeia standard. The comprehensive evaluation index of each plant spacing treatment was 14 cm>16 cm>10 cm>8 cm>12 cm in turn. Conclusion::Combined with the comprehensive evaluation of yield and economic benefit, the optimal transplanting plant spacing and row spacing were 30 cm×14 cm (with the density of 238 100 strains/hm2). Under this density, A. membranaceus var. mongholicus grows vigorously, with thick stems, big root crowns, a high yield and high quality.