Cloning and Expression Patterns of Coumarate-3-hydroxylase Gene from Angelica sinensis and Its Correlation with Content of Ferulic Acid / 中国实验方剂学杂志

ObjectiveTo clone coumarate-3-hydroxylase gene （C3H） from Angelica sinensis， and analyze the correlation between its bioinformatics， expression patterns and content of ferulic acid， and to explore the functions of ASC3H. MethodReal-time polymerase chain reaction （Real-time PCR） was used to clone the full-length cDNA of ASC3H based on the transcriptome dataset of A. sinensis， and the bioinformatics analysis of the gene sequence was carried out. Real-time PCR and high performance liquid chromatography （HPLC） were used to determine relative expression of ASC3H and content of ferulic acid in different root tissues of A. sinensis （periderm， cortex and stele）. ResultThe open reading frame （ORF） of ASC3H （GenBank accession number： MN2550298） was 1 530 bp， encoding 509 amino acids， with a theoretical molecular weight of 57.86 kDa and an isoelectric point of 8.36. It was a hydrophilic protein that was located in the chloroplast with multiple phosphorylation sites and a transmembrane region， and contained a conserved domain CGYDWPKGYGPIINVW_P450 （383-399 aa） in cytochrome P450. Multiple amino acid sequence alignment analysis showed that ASC3H had high similarity with C3H from other plants， especially Ammi majus in Umbelliferae. The Real-time PCR revealed that ASC3H had different expressions in periderm， cortex and stele tissues of A. sinensis roots. It was found from HPLC that the cortex tissues had the highest content of ferulic acid， and the stele tissues had the lowest. ConclusionASC3H was successfully cloned from A. sinensis， and its sequence characteristics were understood more clearly， suggesting that ASC3H might be involved in the ferulic acid biosynthesis pathway of A. sinensis. This paper provided a basis for further studying the functions of the gene and exploring the biosynthesis and regulation mechanism of ferulic acid in A. sinensis， while laying the foundation for the genetic improvement of A. sinensis．

Dimeric phthalides from an aqueous extract of the Angelica sinensis root head / 药学学报

Ten dimeric phthalide racemates (1-10) were isolated from an aqueous extract of the Angelica sinensis root head (Guitou) by separation techniques of column chromatography over macroporous adsorbent resin, MCI resin, silica gel, and Sephadex LH-20, together with preparative thin-layer chromatography and reversed phase HPLC. The racemates were further separated into (+)-/(-)-1-(+)-/(-)-10 with chiral HPLC. Their structures including absolute configurations were elucidated by comprehensive analysis of spectroscopic data, combined with electronic circular dichroism (ECD) and NMR calculations as well as single crystal X-ray diffractions. Compounds (+)-/(-)-1-(+)-/(-)-10 are either new structure or new natural product, named (+)-/(-)-angelidipthalidic acids A-H [(+)-/(-)-1-(+)-/(-)-8] and (+)-/(-)-angelidipthalidols A and B [(+)-/(-)-9 and (+)-/(-)-10], respectively. Meanwhile, dimeric phthalide mono- and bis-lactone derivatives with 3.3′a,8.6′- and 3.6′,8.3′a-coupling patterns as well as determination of their relative configurations are discussed.

Monoterpenes from an aqueous extract of the Angelica sinensis root head / 药学学报

Eleven monoterpenes including seven new chemical structures or new natural products covering two pairs of scalemic enantiomers, together with four known analogues, were isolated from an aqueous extract of the Angelica sinensis root head (Guitou) by separation techniques of column chromatography over macroporous adsorbent resin, MCI resin, silica gel, Sephadex LH-20, and Toyopearl HW-40C, together with preparative thin-layer chromatography as well as reversed phase and chiral HPLC. Their structures were determined by spectroscopic data analysis, combined with theoretic calculation of electronic circular dichroism (ECD) spectra and single crystal X-ray diffraction. The new structures or new natural products named (+)-/(-)-angelinones A and B [(+)-/(-)-1 and (+)-/(-)-2], angelinones C and D (3 and 4), and angelinol A (5), respectively, while the known analogues were 6β,9-dihydroxy-(+)-α-pinene (6), 1,1,5-trimethyl-2-hydroxymethyl-cyclohexa-2,5-dien-4-one (7), jasminol E (8), and (+)-trans-sobrerol (9). All the isolates were reported in this plant for the first time, except for the previously reported 6 from an ethanol extract of the aerial parts of A. sinensis, of which the structure was confirmed by X-ray crystallography in this study.

Angelica sinensis polysaccharides improve Th1/Th2 imbalance and protect pregnancy in threatened abortion model rats / 中国药房

OBJECTIVE To study the tocolysis effects of Angelica sinensis polysaccharides on threatened abortion model rats and their impacts on Th1/Th2 balance by regulating the phosphoinositide 3-kinase （PI3K）/protein kinase B （AKT） signaling pathway. METHODS Pregnant rats were randomly grouped into the control group， model group， A. sinensis polysaccharide group （200 mg/kg）， PI3K/AKT signaling pathway inhibitor LY294002 group （5 mg/kg）， and A. sinensis polysaccharide+LY294002 group （200 mg/kg A. sinensis polysaccharide+5 mg/kg LY294002）， with 10 rats in each group. Except for the control group， rats in all other groups were given mifepristone （8.3 mg/kg） and misoprostol （100 μg/kg） intragastrically to establish a threatened abortion model， and intragastric or intraperitoneal injection of corresponding drugs. The serum levels of estrogen， progesterone， interferon-γ （IFN-γ）， tumor necrosis factor-α （TNF-α）， and interleukin-4 （IL-4） in each group of rats were detected， and the uterine ovarian index and embryonic mortality rate of rats in each group were measured； the morphology of uterine tissue in rats was observed in each group； Th1/Th2 balance in peripheral blood of rats as well as the expression of PI3K/AKT signaling pathway-related proteins in the uterine tissues of rats in each group were detected. RESULTS Compared with the control group， the uterine tissue of rats in the model group showed pathological damage； the serum levels of estrogen， progesterone and IL-4， uterine ovarian index， peripheral blood Th2 cell ratio， and the ratios of phosphorylated PI3K （p-PI3K）/PI3K and phosphorylated AKT （p-AKT）/AKT in uterine tissue were all decreased （P＜0.05）； the embryo mortality rate， Th1 cell ratio， Th1/Th2 ratio， and serum levels of IFN-γ and TNF-α were increased （P＜0.05）. Compared with the model group， the pathological damage of uterine tissue in the A. sinensis polysaccharide group was reduced， and the above indexes were all improved significantly （P＜0.05）； LY294002 could weaken the effect of A. sinensis polysaccharide on model rats （P＜0.05）. CONCLUSIONS A. sinensis polysaccharides can improve Th1/Th2 imbalance in threatened abortion model rats by activating the PI3K/AKT signaling pathway， thereby inhibiting immune inflammation， and promoting embryo survival.

Angelica sinensis polysaccharide promotes stress erythropoiesis in mice caused by 5-FU / 中国药理学通报

Aim To investigate the effects of angelica sinensis polysaccharide (ASP) antagonizing 5-fluorou-raeil (5-FU) on spleen stress erythropoiesis in mice and its related mechanism. Methods C57BL/6J mice aged 6-8 weeks were randomly divided into control group, ASP group, 5-FU group and ASP + 5-FU group. The mouse body weight during the modeling pe-riod was recorded, and peripheral blood routine and the number of mononuclear cells in the bone marrow of femur were measured. Histopathology of spleen was de-tected, also the index and cellularity of spleen were analyzed. BFU-E of spleen mononuclear cells was counted. The number of F4/80

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.

Effects of Angelica sinensis polysaccharide on the apoptosis of cardiomyocytes in diabetic KK-Ay mice / 中国药房

OBJECTIVE To investigate the effects of Angelica sinensis polysaccharide on the apoptosis of cardiomyocytes in diabetic KK-Ay mice. METHODS KK-Ay mice were randomly divided into model group， metformin group （200 mg/kg） and A. sinensis polysaccharide high-dose， medium-dose and low-dose groups （400， 200 and 100 mg/kg）； C57BL/6J mice were included in blank group， with 8 mice in each group. Each group was given relevant medicine intragastrically or normal saline， once a day， for consecutive 4 weeks. After the final administration， the levels of fasting glucose， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C） and insulin （INS） were detected； the protein expressions of B-cell lymphoma 2 （Bcl-2）， cleaved- caspase-3， apoptosis signal-regulated kinase 1 （ASK1）， phosphorylated c-Jun N-terminal kinase （p-JNK）， phosphorylated inositol- requiring enzyme 1α （p-IRE1α） in myocardium， and apoptosis in cardiomyocytes were also detected. RESULTS Compared with model group， the fasting glucose， TC and LDL-C content， apoptotic rate of cardiomyocyte， protein expressions of p-JNK and p- IRE1α， ASK1， cleaved-caspase-3 were significantly lower in the metformin group and A. sinensis polysaccharide medium-dose， high-dose groups； INS level and relative expression of Bcl-2 protein were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS A. sinensis polysaccharide can improve the levels of blood glucose and blood lipid and inhibit cardiomyocyte apoptosis in diabetic KK-Ay mice， and the mechanism may be related to the inhibition of IRE1/ASK1/JNK signaling pathway.

Study on the changes of HPLC fingerprints and main chemical compounds before and after the compatibility of Salvia miltiorrhiza and Salvia miltiorrhiza / 国际中医中药杂志

Objective:To establish methods for HPLC fingerprints and simultaneous determination of multi-index components before and after compatibility of Salvia miltiorrhiza and Angelica sinensis, so as to analyze the dissolution rate of the main compounds. Methods:The extracts of Salvia miltiorrhiza, Angelica sinensis and their compatibility were prepared. The separation was performed on an Eclipse XDB-C18 column (4.6 mm×250 mm,5 μm), mobile phase with 0.1% phosphoric acid aqueous solution-acetonitrile for gradient elution, flow rate at 1.0 ml/min, column temperature was maintained at 35 ℃, and the detection wavelength was set at 280 nm. The HPLC fingerprint were established before and after the compatibility of Salvia miltiorrhiza and Angelica sinensis, and the shared patterns of the fingerprint were obtained to gain chromatographic peaks. The content of 9 components Danshensu, caffeic acid, rosmarinic acid, salvianolic acid B, salvianolic acid A, tanshinone Ⅱ A, ferulic acid, chlorogenic acid and Yangchuanxiong lactone were determinated, and the changes of dissolution rate of each compound before and after the compatibility were analyzed. Results:The determination method for the multi- components with HPLC is precise and the components (waiting to be determinate) in the solution were stable within 48 hours, and the RSD values of each chromatographic peak were <5.0%. The nine components showed good linear relationships within their own ranges, and the recovery rate was in compliance with regulations. The fingerprint similarities of each sample were ?0.9. After the compatibility of Salvia miltiorrhiza and Angelica sinensis, a total of seventeen common peaks were calibrated, ten of which were from Salvia miltiorrhiza, seven from Angelica sinensis. No new components was found under this chromatographic condition. After the combination of these two material medicica decoction, the average dissolution rates of rosmarinic acid, salvianolic acids and Danshensu in Salvia miltiorrhiza were significantly lower than those of the single decoction group ( P<0.05 or P<0.01); the average dissolution rates of caffeic acid in Salvia miltiorrhiza was significantly higher than that of the single decoction group ( P<0.01); the average dissolution rates of chlorogenic acid and ferulic acid in Angelica sinensis were significantly higher than that of the single decoction group ( P<0.05 or P<0.01); the average dissolution rate of Yangchuanxiong lactone after the compatibility was not statistically different than that of single decoction group. Conclusion:The characteristic peaks of HPLC fingerprint of the compatibility of Salvia miltiorrhiza and Angelica sinensis did not increase under this chromatographic condition, which had a significant effect on the dissolution of index components.

Rapid identification of phthalides and organic acids in Angelica sinensis based on UPLC-Q-TOF/MS / 中国药房

OBJECTIVE To establish a comprehensive and rapid m ethod for the a nalysis of chemical constituents as phthalides and organic acids in Angelica sinensis ，and to provide scientific reference for the quality evaluation and pharmacodynamic substance research of A. sinensis . METHODS The 70％ ethanol extract of A. sinensis was analyzed by ultra high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry （UPLC-Q-TOF/MS）. The determination was performed on ACQUITY UPLC BEH-C 18 column with mobile phase consisted of 0.1％ formic acid solution- acetonitrile（gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was set at 30 ℃，and sample size was 2 µL. The ion source was an electrospray ion source ，using positive ion scanning mode ，and the mass scanning range was m/z 50-1 000. Capillary voltage was 4 000 V； atomizer pressure was 35 psi；cracking voltage was 135 V and the taper hole voltage was 65 V；the temperature of dry gas was 320 ℃；the flow of dry gas was 10 L/min and the flow of sheath gas was 11 L/min；collision energy were 20 and 40 V. Qualitative Analysis 10.0 software was used to obtain the retention time of compounds ，the accurate mass number of excimer ion peaks and secondary fragments. The compounds were analyzed by comparing with the mass spectra of the reference substance ，combined with relevant literature ，mass spectrometry cleavage law and database such as Chemspider ，MassBank，PubChem. RESULTS A total of 72 compounds were identified or deduced from A. sinensis ，including 55 phthalides，13 organic acids and 4 other constituents. CONCLUSIONS The established method is rapid and accurate for the identification of chemical constituents from A. sinensis ，such as organic acids and phthalides ，which provides an efficient and rapid analytical method for the comprehensive characterization of its chemical constituents.

Identification and Genetic Variation Analysis of Wild and Cultivated Germplasm of Angelica sinensis Based on cp DNA / 中国实验方剂学杂志

ObjectiveTo conduct genetic variation analysis of 11 cultivars and 7 wild populations of Angelica sinensis in Gansu province based on the chloroplast gene （cp DNA）， and provide references for germplasm identification and breeding of new cultivars of A. sinensis. MethodThree pairs of cp DNA primers were used for polymerase chain reaction （PCR） amplification and sequencing of A. sinensis samples. MegaX was used to perform statistics on sequence characteristics and calculate mean genetic distances among A. sinensis populations. Unweighted pair-group method with arithmetic means （UPGMA） clustering tree based on genetic distance was constructed by NTSYS 2.10e. DanSP v6 was used to calculate sequence polymorphism and Tajima's D of A. sinensis. PERMUT was used to calculate the population structure of A. sinensis. Arlequin v3.5 was used to perform molecular variation analysis， and PopART1.7 was used to construct TCS haplotype network. ResultThree pairs of cp DNA primers were amplified， sequenced， compared， and combined to give a sequence length of 1 759 bp. One variable site was detected in the wild A. sinensis and 480 variable sites were detected in the cultivated A. sinensis， including 97 singleton variable sites， 383 parsimony informative sites， and 152 insertion-deletion sites. In the three regions of matK， psbA-trnH， and rbcL of cp DNA in the wild and cultivated A. sinensis， matK was the region with the highest polymorphism. Tajima’s D of all the combined sequences of A. sinensis were not significantly negative， but psbA-trnH and rbcL genes of the cultivated A. sinensis were significantly negative， indicating that the A. sinensis followed neutral evolution on a whole， while psbA-trnH and rbcL genes had undergone selection. The degree of genetic differentiation （Fst=0） among wild populations was lower than that among cultivated populations （Fst=0.114 19， P<0.05）. The degree of genetic differentiation between wild and cultivated A. sinensis was relatively high （Fst=0.942 55， P<0.01）. Genetic variation in the cultivated A. sinensis was mainly found within the populations （89%）. UPGMA cluster tree based on genetic distance showed that the wild A. sinensis and the cultivated A. sinensis were clustered into one branch， respectively， with a distant genetic relationship， and the population 3 in the cultivated A. sinensis was far from other cultivated populations. The TCS haplotype network consisted of 15 haplotypes and 4 unknown haplotypes， which was divided into 3 parts， with a large number of variations among each part. Shared haplotypes were only found in the wild or cultivated groups， and there were no shared haplotypes between groups. ConclusionThe genetic diversity of A. sinensis was low at species level， and the population diversity of the wild was lower than that of the cultivated. The degree of genetic differentiation between the wild and the cultivated A. sinensis was high， but that in the wild and the cultivated populations were low. Genetic variation in the cultivated A. sinensis was mainly found within populations.

Effect of Micro-fertilizer Containing Rare Earth on Growth，Yield and Quality of Angelica sinensis / 中国实验方剂学杂志

ObjectiveTo investigate the effects of micro-fertilizer containing rare earth of different types and concentrations on the growth，yield and quality of Angelica sinensis. MethodOn the basis of the single-factor randomized block design， the growth and index components of Angelica sinensis were determined with rare earth-containing nitrate and chloride micro-fertilizers of different concentrations as foliar fertilizers. ResultSpraying 0.8 g·mL-1 rare earth-containing chloride micro-fertilizer could increase the economic yield of A. sinensis， with the fresh yield per mu （1 mu≈667 m2） reaching 855.4 kg and the dry yield per mu 350.7 kg，which increased by 15.16% and 28.70% respectively compared with that in the control group CK1. Spraying 1.2 g·mL-1 rare earth-containing nitrate micro-fertilizer could promote the growth and development of A. sinensis and significantly increase the content of index components， with the plant height reaching 93.05 cm，the stem diameter 15.60 mm，the root diameter 16.10 mm，the main root length 36.5 cm，and the number of leaves 11.25 pieces per plant， which increased by 32.76%，31.98%，41.98%，53.36%，and 45.16%， respectively， compared with those in the control group CK2. Besides， the content of ferulic acid，volatile oil，ligustilide， and extract was 0.96%，0.41%，0.30% and 48.76%，respectively，which increased by 12.94%，17.14%，11.11%， and 12.07%，respectively，compared with that in the control group CK2. ConclusionSpraying 0.8 g·mL-1 rare earth-containing chloride micro-fertilizer and 1.2 g·mL-1 rare earth-containing nitrate micro-fertilizer can promote the growth and development of A. sinensis，improve the medicinal properties，and increase yield and quality. Rare earth-containing micro-fertilizers can be applied in the standardization of A. sinensis cultivation， which can change the production status of A. sinensis that depends on chemical fertilizers and single fertilization， and promote the green， organic and ecological cultivation of A. sinensis.

Interventional effect of Radix Angelica Sinensis and Radix Hedysari ultrafiltration on miR-21-5P targeting TGF-β1-mediated radiation-induced myocardial fibrosis / 中国临床药理学与治疗学

AIM: To reveal that the targeted regulation of TGF-β1 by miR-21-5P is the key mechanism that mediates the activation of myocardial fibroblasts, and to clarify the intervention ofRadix Angelica Sinensis and Radix Hedysari ultrafiltration on the mechanism of miR-21-5P targeting to regulate TGF-β1 effect. METHODS: (1) The cells were randomly divided into normal group and irradiation group. The irradiation group received 6Gry single irradiation, and then RT-PCR was used to detect miR-21-5P, and Western Blot was used to detect the expression of α-SMA and TGF-β1. (2) The cells were randomly divided into normal group, irradiation group, miR-21-5P

Exploration of mechanism of Huoxue Jiedu Chinese Herbal Compatibility in anti-atherosclerosis: Based on network pharmacology and molecular docking / 中国药理学通报

Aim To explore the mechanism of Huoxue Jiedu Chinese Herbal Compatibility (angelica sinensis - lonicerae japonicae flos ) in anti-atherosclerosis by means of network pharmacology and molecular rloe- king.Methods The effective eomponents and related targets of angelica sinensis and lonicerae japonicae flos were sereened by the TCMSP database, and the dis¬ease related targets of atheroselerosis were obtained by using the Gene Cards database, DrugBank database and OMIM database.The common targets were ob¬tained by Venny2.1 , an online mapping tool.The pro- tein-protein interaction ( PPI) network was established by String 11.0, and the common target was analyzed by gene ontology(GO) enrichment analysis and kvoto en- cyelopedia of genes and genomes ( KEGG) enrichment analysis using metascape online platform.Cyto- seape3.8.2 was used to construct the " active ingredi¬ent-active target-pathway network map.Finally AutoDock and PyMOL softwares were applied for mo¬lecular docking.Results A total of 31 active compo¬nents of angelica sinensis-lonicerae japonicae flos com¬ patibility and 509 component targets were screened.Hie number of intersection targets with atherosclerosis was 210.GO function enrichment analysis revealed 2 961 biological process items and 242 molecular func¬tion items.KEGG pathway enrichment screened 250 signaling pathways, PI3K-Akt signaling pathway, MAPK signaling pathway, and fluid shear stress and atherosclerosis were the key pathways.Molecular doc¬king results showed that beta-sitosterol, ferulic acid, rutin, luteolin, quercetin could bind stably with MAPK (-1,-3), AKT1, PRKC ( A, B).Conclusions Huoxue Jiedu Chinese Herbal Compatibility ( angelica sinensis -lonicerae japonicae flos) containing multiple active ingredients may play an anti-atherosclerosis role through the signaling pathways of MAPK, P13K-Akt and PRKC, reflecting the advantages of the treatment of activating blood circulation and detoxification of tra¬ditional Chinese medicine in anti-atherosclerosis.

Effect of Garlic on Soil Environment and Yield of Continuous Cropping and Rotation of Angelica sinensis Under Membrane Cultivation / 中国实验方剂学杂志

Objective:To study the change of soil environment and yield of <italic>Angelica sinensis </italic> when cultivated on the film side and and open field sides，and to explore the influence of garlic volatiles and film-side cultivation on continuous cropping <italic>A. sinensis</italic>，so as to provide some certain theoretical basis for the saying that the <italic>A. sinensis</italic> garlic intercropping mode and film-side cultivation could alleviate continuous cropping <italic>A. sinensis</italic>. Method:In this study， the effects of garlic on soil environment and yield of <italic>A. sinensis</italic> cultivated on film side and crop rotation were discussed through the determination and analysis of soil microorganism quantity，soil enzyme activity and yield of Angelica sinensis in the rhizosphere of <italic>A. sinensis</italic>. Result:The number of bacteria and actinomycetes in the rhizosphere soil of <italic>A. sinensis </italic>increased significantly， while the number of fungi decreased significantly. Among them， the number of bacteria and actinomycetes in the intercropping of <italic>A. sinensis</italic> and garlic on the film side increased by 104% and 146.89% on average as compared with that of the control （monoculture under continuous cropping open field），and the number of fungi decreased by 39.28% on average as compared with that of control group. When the enzyme activity in the rhizosphere soil was significantly increased under the condition of intercropping with garlic on the film side，the soil urease，sucrase，alkaline phosphatase and acid phosphatase activities under the intercropping of <italic>A. sinensis</italic> and gralic on the rotation film side were increased by 61.60%，51.09%，203.48% and 76.19% respectively as compared with those in control group. The yield of <italic>A. sinensis</italic> significantly increased under the condition of intercropping with garlic on the film-side，and the yield of Angelica sinensis under the rotation film-side intercropping with garlic increased by 39.37% as compared with that of control group. Conclusion:Under the mode of film-side cultivation，the interplanting of <italic>A. sinensis </italic>with garlic can change the number of soil microorganism，improve the activity of soil enzyme，improve the physical and chemical properties of soil， alleviate the obstacle of continuous cropping and guarantee the healthy growth of <italic>A. sinensis</italic> more effectively.

Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP / 中国实验方剂学杂志

Objective:To establish a rapid method to identify <italic>Levisticum officinale </italic>adulterated in<italic> Angelica sinensis</italic> by polymerase chain reaction -restriction fragment length polymophism（PCR-RFLP）. Method:By comparing sequences restriction sites in ribosomal DNA Internal Transcribed Spacer（ITS） of <italic>A. sinensis</italic> and <italic>L. officinale</italic>，the specific restriction site Fnu4HI of <italic>L. officinale</italic> was selected，and the primers for PCR-RFLP reaction were designed. Different <italic>A. sinensis</italic> and <italic>L. officinale</italic> were amplified by PCR. The conditions affecting the PCR-RFLP reaction，such as annealing temperature，primer concentration，cycle number and enzyme digestion reaction time，were optimized，and the accuracy of the method was investigated. The established PCR-RFLP identification method was used to investigate the applicability of <italic>L. officinale </italic>adulterated<italic> in A. sinensis</italic> with different aduleration ratios and different origins. Result:A PCR-RFLP method for identifying <italic>A. sinensis</italic> mixed with <italic>L. officinale</italic> was established. When the annealing temperature was 62 ℃ and the number of cycles was 30，when the <italic>L. officinale </italic>adulterated in<italic> A. sinensis</italic> could be digested by Fnu4H I restriction endonuclease after amplification with specific primers，and the two single DNA bands were detected between 100-500 bp，the <italic>A. sinensis</italic> were all negative. The minimum detection limit of this method for adulterated <italic>L. officinale</italic> in <italic>A. sinensis</italic> was 3%，which could be used for the detection of adulterated <italic>L. officinale</italic> in <italic>A. sinensis</italic>. Conclusion:The established PCR-RFLP identification method is sensitive and accurate in detecting whether there is <italic>L. officinale</italic> in <italic>A. sinensis</italic>，and it provides inspection reference and basis for the quality control of <italic>A. sinensis</italic>，with great significance to ensure the safety of its clinical medication.

Quality Regionalization of Angelica sinensis in Genuine Producing Area / 中国实验方剂学杂志

Objective:To study on the quality regionalization of <italic>Angelica sinensis</italic>，in order to guide the rational cultivation of <italic>A. sinensis</italic>. Method:Through visits and field surveys，a total of 857 batches of <italic>A. sinensis</italic> were collected from different counties of Dingxi，relevant geographic information such as longitude，latitude，altitude of each sampling point were obtained by using the global positioning system（GPS），the content of 8 indexes in <italic>A. sinensis </italic>was detected by UPLC， and based on national ecological environment factor data，the suitability analysis of the quality of <italic>A. sinensis</italic> was performed by using MaxEnt，ArcGIS，SPSS. Result:The suitable areas of <italic>A. sinens </italic>were concentrated in central and southern Dingxi. In the suitable areas，the content of ferulic acid，coniferyl ferulate，senkyunolide H decreased from south to north，the content of chlorogenic acid decreased from north to south，the content of senkyunolide A，senkyunolide I decreased from east to west，the distribution regularity of butenyl phthalide was not strong，the highest-content areas were in western Min county，Qingyuan town of Qingyuan county，Shangwan Township and Huichuan town. The content of ligustilide was consistent in the suitable area，and the highest content were in the middle of Weiyuan county and the northern Tongwei county. The results showed that it had a higher index components and comprehensive quality in Min county，Zhang county，southern Weiyuan county and northern Tongwei county. Conclusion:In this study，the quality suitability areas of <italic>A. sinensis</italic> in Dingxi were graded. The chemical components and the quality suitability zoning maps were generated. The findings could provide references for the comprehensive utilization of <italic>A. sinensis</italic>，the selection and construction of high-quality <italic>A. sinensis</italic> raw material base，and the scientific guidance for the production and regional development of <italic>A. sinensis</italic> in Dingxi.

Development and application of a chemical transformational method for the indirect measurement of ligustilide content in Angelica sinensis / 药学学报

The poor stability of the ligustilide (LIG) makes its quantitation in Angelica sinensis (AS) difficult. This study establishes a chemical conversion method for the determination of ligustilide content in AS and proposes a national pharmacopoeia standard. Mechanical agitation and sonication of a powdered AS extract in a methanol/cyprolamine mixture facilitated the stabilization and transformation of ligustilide. Using an external reference HPLC-DAD method, the cyclopropyl-ligustilide (LIGc) content in the mixture could be determined. The content of ligustilide was greater than 1.0% based on 144 AS specimens including 68 obtained from the originally planted areas of Qinghai and Gansu Province; 55 specimens were obtained from Minxian and Weiyuan County medicine markets, and 21 specimens for which the storage period reached or exceeded 1.5 years. According to the Hong Kong Chinese materis medica standards, the content of ligustilide in AS should not be lower than 0.6%. The developed method could also be applied to the quality control of other Chinese medicinal materials (such as Ligusticum chuanxiong) or Chinese patent medicines in which ligustilide is the main component.

Effects of Angelica Sinensis polysaccharide on proliferation in vitro and transplantation of human leukemia stem cells in vivo / 解剖学报

Objective To investigate the effect of Angelica Sinensis polysaccharide (ASP) on proliferation, differentiation and transplantation of human leukemia stem cells (LSCs) . Methods 1. Effect of angelica sinensis polysaccharides on proliferation of CD34

Heavy Metals in Danggui (Angelica sinensis) Consumed by Postpartum Mothers and Its Health Risk

@#Introduction: Consumption of Chinese Herbal Medicine (CHMs) have escalated globally. They are preferred treatment for minor diseases or disorders. In Malaysia, CHMs are common home remedies during pregnancy and postpartum. Angelica sinensis (Danggui) is a staple CHMs during postpartum for purpose of nourishing blood and resolving stasis. Concerns are raised over possible heavy metals toxicity. Objective: This study aims to (i) determine Danggui consumption among postpartum mothers, (ii) quantify its heavy metals level, namely Lead (Pb), Cadmium (Cd), Arsenic (As) and Chromium (Cr) and (iii) determine health risks of Danggui consumption among mothers. Methods: A cross-sectional study involving 112 postpartum mothers was carried out in Kuala Lumpur. Danggui samples were collected from nine districts in Kuala Lumpur (Segambut, Seputeh, Cheras, Kepong, Bandar Tun Razak, Titiwangsa, Setiawangsa, Batu and Lembah Pantai). Heavy metals were extracted using microwave digester and analysed using Inductively coupled plasma mass spectrometry (ICPMS). Hazard Quotient (HQ) was used to determine non-carcinogenic health risks for herbal medicine consumption. Results: Danggui was consumed by 19.6% of mothers (n=22). Among them, incidence of jaundice was 63.6% and need for phototherapy was 40.9%. Heavy metals contaminations were found in the decreasing order of Cr > As > Pb > Cd with median (interquartile) of 3996.3 (2805.6) μg/ kg, 128.3 (56.7), 98.6 (99.1) and 37.0 (35.0) respectively. No non-carcinogenic health risks were found for all four metals. Conclusion: Alarming concentrations of heavy metals were quantified in Danggui warranting for further investigation to safeguard health of postpartum mothers.

Metabonomics Analysis on Different Varieties of Angelica sinensis Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective::To establish differential metabolites between different varieties of Angelica sinensis, and provide reference for breeding, introduction, regional cultivation and ecological cultivation of new varieties of A. sinensis. Method::Comprehensive non-target metabonomics analysis was conducted for five new varieties of A. sinensis collected at the same time from the same origin: Mingui No. 1 (MG1), Mingui No. 2 (MG2), Mingui No. 4 (MG4), Mingui No. 5 (MG5), and Mingui No. 6 (MG6). The 50% methanol extract of each variety was taken, and then the differential metabolites among varieties were found by using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), software Progenesis QI, principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and non-targeted metabonomics analysis. Differential metabolites were identified based on precise molecular weight, secondary fragments, KEGG database, HMDE database and related literature information. Result::The results of UPLC-Q-TOF-MS and multivariate statistical analysis showed that there were significant differences in the metabolites of five Angelica varieties. As compared with MG1, the contents of chlorogenic acid, ferulic acid, tryptophan and ferulic aldehyde were significantly lower in MG2, MG4, MG5 and MG6, while the contents of ligustilide, coumarin, bovine keratin, palmitin, protocatechualdehyde and linolenic acid were significantly higher (P<0.05). The results of multivariate statistical analysis showed that the metabolites of MG2 and MG5 were similar with those of MG6, but were significantly different from those of MG4.In addition, 38 distinct metabolites were identified, involving 7 potential targeted metabolic pathways. Different varieties of A. sinensis could regulate the synthesis of their metabolites through phenylpropanoid biosynthesis and sesquiterpene-like compounds metabolism, lipid metabolism, amino acid metabolism, carbohydrate metabolism, nucleotide metabolism, carotenoids, linolenic acid, linoleic acid and some other metabolic pathways. Conclusion::UPLC-Q-TOF-MS and Progenesis QI metabonomics techniques were used to compare the chemical constituents of different varieties of A. sinensis from the overall level. The differences and their regularities were found, which could provide reference for quality control, variety sorting, identification, breeding and ecological planting of A. sinensis.