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Issue 14 ， 2021
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Antidepressant Effect of
n -Butyl Alcohol Extracts from Xiaoyaosan via IGF-1Rβ /PI3K/Akt PathwayAbstract：ObjectiveTo investigate the intervention effect of n-butyl alcohol extracts from Xiaoyaosan against depression-like behavior induced by chronic unpredictable mild stress （CUMS） in model mice and the role of insulin-like growth factor-1 receptor β （IGF-1Rβ）/phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt） signaling pathway in such intervention.MethodThe effective dose of n-butyl alcohol extracts from Xiaoyaosan was preliminarily determined in model mice with behavioral despair. Then the male C57BL/6 mice were randomly divided into the blank group， model group， fluoxetine group， Xiaoyaosan group， and the low- （20 g·kg-1） and high-dose （40 g·kg-1） n-butyl alcohol extract groups. The mice in all groups except for the blank group were exposed to CUMS for inducing the depression-like behavior， which was judged by the sucrose preference test （SPT）. The successfully modeled mice in the medication groups were intragastrically administered with the corresponding drugs， whereas those in the blank and model groups were treated with an equal volume of solvent for five successive weeks. Following the SPT， tail suspension test （TST）， and novelty suppressed feeding test （NSFT） at the end of the fifth week， the insulin-like growth factor-1 （IGF-1） levels in mouse serum and hippocampus were measured by enzyme-linked immunosorbent assay （ELISA）. The average optical density （IA） of Nissl bodies in mouse hippocampal CA3 region was detected by toluidine blue staining. The 5-bromo-2-deoxyuridine （Brdu） and doublecortin （DCX） expression in the dentate gyrus （DG） was assayed using immunofluorescence method. The protein expression levels of IGF-1Rβ， PI3K， phosphorylated-PI3K （p-PI3K）， Akt， p-Akt， cysteine aspartic acid-specific protease 3 （Caspase-3）， and cleaved Caspase-3 in the hippocampus were determined by Western blot.ResultThe results of forced swimming test and TST showed that n-butyl alcohol extracts from Xiaoyaosan at 9.1 and 40 g·kg-1 both significantly shortened the immobility time of mice （P<0.05， P<0.01）， indicating that the effective dose ranged from 9.1-40 g·kg-1. Compared with the model control， the n-butyl alcohol extracts from Xiaoyaosan at 20 and 40 g·kg-1 significantly increased the sucrose preference percentage （P<0.05， P<0.01）， shortened the immobility time in TST （P<0.01） and the feeding latency in NSFT （P<0.01）， reversed the down-regulated IGF-1 content in mouse serum and hippocampus （P<0.01）， increased the AOD of Nissl bodies in the hippocampal CA3 region （P<0.01）， promoted the expression of Brdu and DCX in DG （P<0.05， P<0.01）， and down-regulated the protein expression levels of IGF-1Rβ （P<0.05， P<0.01）， p-PI3K/PI3K （P<0.05， P<0.01）， p-Akt/Akt （P<0.05）， and cleaved Caspase-3/Caspase-3 in the hippocampus of CUMS mice.ConclusionThe n-butyl alcohol extracts from Xiaoyaosan are equivalent to Xiaoyaosan in inhibiting expression. They alleviate the depression-like behavior in CUMS mice， induce the production of Nissl bodies in hippocampal CA3 region， enhance neuronal proliferation and differentiation in DG， and facilitate neurogenesis. All these may be related to the inhibition of over-activated IGF-1Rβ/PI3K/Akt pathway and the reduction of neuronal apoptosis.Keywords：Xiaoyaosan;n-butanol fraction;chronic unpredictable mild stress （CUMS）;neurogenesis;apoptosis;insulin-like growth factor-1 receptor β （IGF-1Rβ）/phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt） signaling pathway- 41
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Analysis on UPLC Fingerprint and Determination of Multi-index Components in Pipa Qingfeiyin
Abstract：ObjectiveTo establish the ultraperformance liquid chromatography （UPLC） fingerprint of Pipa Qingfeiyin substance benchmark， and to establish a quantitative analysis method for simultaneous determination of the contents of five index components， so as to provide reference for the quality control and evaluation of this famous classical formula.MethodACQUITY UPLC® CSHTM C18 column （2.1 mm×100 mm， 1.7 μm） was used with mobile phase of acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-7 min， 5%-7%A； 7-11 min， 7%-8%A； 11-22 min， 8%-14%A； 22-30 min， 14%-15%A； 30-35 min， 15%-25%A； 35-42 min， 25%-40%A； 42-45 min， 40%-50%A； 45-50 min， 50%-60%A）， the flow rate was 0.35 mL·min-1， the column temperature was 25 ℃， the detection wavelengths were 278 nm and 248 nm. UPLC fingerprints of 15 batches of Pipa Qingfeiyin substance benchmark were established， and the "Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine" software （2012 edition） was used for similarity analysis， and the common peaks were assigned. Cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used to evaluate the fingerprint data. UPLC fingerprint method was used to simultaneously determine the contents of five components in the substance benchmark.ResultThe method validation of fingerprint and determination method was good， the similarities between 15 batches of Pipa Qingfeiyin substance benchmark and their control fingerprint were ≥0.997， 23 common peaks were identified and 11 chromatographic peaks were identified. CA， PCA and OPLS-DA divided 15 batches of the substance benchmark into two groups. The linear relationship of phellodendrine hydrochloride， chlorogenic acid， berberine hydrochloride， palmatine hydrochloride and ammonium glycyrrhizinate was good in a certain range of concentration （R2>0.999）， their average recovery was 96.47%-101.16%， and the contents of these five components in the substance benchmark were 0.87-2.00， 1.53-5.95， 18.45-33.97， 3.87-6.29， 1.02-4.12 mg·g-1， respectively.ConclusionThe established UPLC fingerprint and multi-index component content determination methods have strong specificity， good resolution and high sensitivity， it can be characterized except for the Ginseng Radix et Rhizoma flavor， which can provide reference for the quality control and evaluation of Pipa Qingfeiyin compound preparation.Keywords：famous classical formulas;Pipa Qingfeiyin;substance benchmark;fingerprint;determination;cluster analysis （CA）;principal component analysis （PCA）;orthogonal partial least squares discriminant analysis （OPLS-DA）- 33
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Inhibitory Effect of Sangsuyin on Airway Inflammation in Asthmatic Rats by Regulating TLR4 Pathway
Abstract：ObjectiveTo investigate the inhibitory effect of Sangsuyin on airway inflammation in asthmatic rats by regulating the Toll-like receptor 4 （TLR4） pathway.MethodForty-eight SD rats were randomly divided into a normal group （n=8） and an experimental group （n=40）. Asthma model was induced in the rats of the experimental group which were further divided into the following five groups according to a random number table： a model group， a dexamethasone group （0.005 g·kg-1），and low- （2.1 g·kg-1）， medium- （4.2 g·kg-1）， and high-dose （8.4 g·kg-1） Sangsuyin groups. The drugs were all dissolved in normal saline at 0.01 L·kg-1. The rats in the model group and the normal group received normal saline （ig） at 0.01 L·kg-1. The drug treatment was carried out once per day， for a total of 7 days. The grades of allergic reactions were compared among the groups after intervention. The levels of interleukin-1β （IL-1β），tumor necrosis factor-α（TNF-α）， and interferon-γ（IFN-γ） in serum and bronchoalveolar lavage fluid （BALF） were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of lung tissues，and the inflammatory cell infiltration scores of lung tissues were compared. The mRNA expression of TLR4 and nuclear factor-κB （NF-κB） was detected by quantitative real-time fluorescence-based reverse-transcription polymerase chain reaction （Real-time PCR）. The expression of TLR4，NF-κB， and the level of phosphorylated NF-κB （p-NF-κB） were detected by Western blot.ResultThe success rate of modeling was 95.00%. Compared with the normal group， the model group showed a higher grade of allergic reaction and more severe pathological changes of lung tissues，and the groups with drug intervention exhibited relieved conditions. The levels of IL-1β and TNF-α in the serum and BALF were higher in the model group than in the normal group，and lower in the groups with drug intervention than in the model group （P<0.01）. The level of IFN-γ in the serum and BALF was lower in the model group than in the normal group，and higher in the groups with drug intervention than in the model group （P<0.05，P<0.01）. The inflammatory cell infiltration score，mRNA and protein expression of TLR4 and NF-κB，and the mRNA level of p-NF-κB were higher in the model group than in the normal group，and lower in the groups with drug intervention than in the model group （P<0.01）.ConclusionSangsuyin could inhibit allergic reactions，lung tissue lesions， and airway inflammation in asthmatic rats. It is speculated that this effect is achieved by inhibiting the TLR4 pathway，down-regulating the mRNA and protein expression of TLR4 and NF-κB，and reducing the level of p-NF-κB.angsuyin can inhibit allergic reaction，lung tissue lesions and airway inflammation in asthmatic rats. It is speculated that this effect is achieved by inhibiting TLR4 pathway，down regulating the mRNA and protein expression of TLR4，NF-κB，and reducing the levels of p-NF-κB.Keywords：Sangsuyin;Toll-like receptor 4 （TLR4）;nuclear factor-κB （NF-κB）;mitogen-activated protein kinase （MAPK）;asthma;airway inflammationangsuyin- 24
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Classic Prescriptions
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Inhibitory Effect and Mechanism of Jingulian Extract on LPS-induced RAW264.7 Cell Inflammatory Response Based on PI3K/Akt Signaling Pathway
Abstract：ObjectiveTo explore the inhibitory effect and mechanism of Jingulian extract （JGL） on inflammation.MethodThe following groups were set up in this study： a control group （10% fetal bovine serum）， a lipopolysaccharide （LPS） model group （0.5 mg·L-1）， and JGL groups （10， 20， 40， 60， 80， 120， 160， 200， 250， 300 mg·L-1 + 0.5 mg·L-1 LPS）. The RAW264.7 cells were cultured for 24 hours. Cell proliferation was detected by cell counting kit-8 （CCK-8） assay. Nitric oxide （NO） release was detected by Griess assay. The release of cytokines interleukin （IL）-1β， IL-6， IL-10， and tumor necrosis factor （TNF）-α was determined by enzyme linked immunosorbent assay （ELISA）. The expression of inducible nitric oxide synthase （iNOS） and intraprostaglandin peroxidase synthase 2 （PTGS2）/cyclooxygenase-2 （COX-2） was measured by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and the activation of key proteins in the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway by Western blot.ResultCompared with the control group， LPS （0.5 mg·L-1）could promote the proliferation of RAW264.7 cells after stimulation for 24 hours （P<0.01）. Compared with the model group， JGL had no significant effect on cell proliferation. Compared with the control group， LPS （0.5 mg·L-1）increased the release of NO， IL-1β， IL-6， IL-10， and TNF-α （P<0.01）. Compared with the model group， JGL （20-300 mg·L-1）inhibited the release of NO in a dose-dependent manner after stimulation for 24 hours （P<0.05） and reduced IL-1β， IL-6， and IL-10 （P<0.05， P<0.01）， but no obvious inhibition on the release of TNF-α was observed. LPS （0.5 mg·L-1） could induce the expression of iNOS and PTGS2/COX-2 genes as compared with the control group （P<0.05， P<0.01）. JGL could down-regulate the mRNA expression of iNOS and PTGS2/COX-2 genes as compared with the model group （P<0.05， P<0.01）. LPS （0.5 mg·L-1） could activate the PI3K/Akt pathway （P<0.01） as compared with the control group， while JGL （10， 20， 40， and 80 mg·L-1） decreased the expression of PI3K-p110， p-p85， and p-Akt （P<0.01）， and inhibited the activation of PI3K/Akt pathway.ConclusionJGL extract could significantly inhibit the inflammatory response and activation of the PI3K/Akt pathway induced by LPS in RAW264.7 cells. The anti-inflammatory effect was related to the inhibition of the PI3K/Akt pathway.Keywords：Jingulian extract （JGL）;lipopolysaccharide （LPS）;RAW264.7 cell;inflammation;phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway- 34
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Effect of
Usnea diffracta Against Atherosclerosis in Rats and on Microbial Flora of Ileum： An Investigation Based on Interior-exterior Relationship Between Heart and Small IntestineAbstract：ObjectiveTo explore the correlation between the efficacy of Usnea diffracta in treating atherosclerosis （AS） and the altered microbial flora in rat ileum based on the interior-exterior relationship between heart and small intestine in traditional Chinese medicine （TCM）.MethodForty-eight SD rats were randomized into a normal group （n=8） and a modeling group （n=40）. The AS model was established with high-fat diet combined with intraperitoneal injection of vitamin D3. The successfully modeled rats were further randomly divided into the model group， positive control （simvastatin， 4 mg·kg-1） group， and low- （0.7 g·kg-1）， medium- （1.4 g·kg-1）， and high-dose （2.8 g·kg-1） U. diffracta ethanol extract groups， with eight rats in each group. After four weeks of intervention， the blood， aorta， ileum， and ileum content of rats in each group were collected. The levels of serum lipopolysaccharide （LPS）， tumor necrosis factor-α （TNF-α） and interleukin 6 （IL-6） were measured by enzyme-linked immunosorbent assay （ELISA）， and the pathological changes in rat thoracic aorta was detected by hematoxylin-eosin （HE） staining. Western blot was conducted to determine the protein expression levels of tight junction protein zonula occluden （ZO-1） and Occludin in rat ileum， and the high-throughput 16S rRNA sequencing technology was employed to detect changes in microbial diversity and abundance in rat ileum of each group.ResultCompared with the normal group， the model group exhibited obvious aortic plaque deposition， increased LPS， TNF-α， and IL-6 levels （P<0.01）， but decreased ZO-1 and Occludin protein expression （P<0.01）. The comparison with the model group revealed that U. diffracta significantly ameliorated the aortic plaque deposition of model rats， lowered serum LPS， TNF-α， and IL-6 levels （P<0.05， P<0.01）， and up-regulated ZO-1 and Occludin protein expression （P<0.05， P<0.01）. The abundance of Proteobacteria and Bacteroidetes in the model group changed significantly in contrast to that in the normal group， and the Bacteroidetes/Firmicutes（B/F） value declined （P<0.05）. Alpha and Beta diversity analysis indicated higher total number of intestinal flora species in the model group， but lower richness and uneven distribution （P<0.05， P<0.01）， with a large number of pathogenic bacteria enriched. The ethanol extract of U. diffracta significantly increased the B/F value， corrected the structural disorder of microbial flora in ileum， reduced pathogenic bacteria， and increased the relative abundance of probiotics.ConclusionU. diffracta exerts the therapeutic effect against AS possibly by improving the intestinal microbial communities， strengthening the intestinal mucosal barrier function， and reducing the serum LPS and inflammatory factors.Keywords：Usnea diffracta;atherosclerosis （AS）;inflammation;intestinal mucosal barrier;intestinal flora- 23
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Regulation of Hypnotic Active Components of Cnidii Fructus on Melatonin Synthesis Rate-limiting Enzyme AANAT in Rats with PCPA-induced Insomnia
Abstract：ObjectiveTo observe the effects of Cnidii Fructus hypnotic active components （CHC） on the behaviors of rats with p-chlorophenylalanine （PCPA）-induced insomnia and melatonin （MT） synthesis rate-limiting enzyme arylalkylamine N-acetyltransferase （AANAT）， and explore the protective mechanism of CHC on the pineal gland.MethodMale SD rats of SPF grade were randomly divided into a blank control group， a model group， a MT group， and high-， medium-， and low-dose CHC groups with 10 rats in each group. Except for the blank control group， other groups received 4.5% PCPA suspension at 10 mL·kg-1， intragastric administration， for two consecutive days. After PCPA model of insomnia was established， normal and model groups were gavaged at the same volume of 2% Tween-80， MT control group （10 mg·kg-1）， CHC was high， medium and low （60， 30， 15 mg·kg-1）， 10 mL·kg-1， once a day， for consecutive 7 days. Four days after administration， open field， elevated cross maze， and pentobarbital sodium-induced sleep tests were conducted， respectively. Serum MT was detected by enzyme-linked immunosorbent assay. The mRNA expression level of AANAT was determined by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）. The expression of AANAT protein in the pineal gland was detected by Western blot.ResultCompared with the results in the blank control group， the total distance of open field activity and standing times and duration in the central area were increased （P<0.05， P<0.01）， the proportions of open arm entry （OE%） and open arm time （OT%） were decreased （P<0.05）， and the sleep latency was prolonged （P<0.01） in the model group. Compared with the model group， no significant difference was observed in the low-dose CHC group， while other groups exhibited reduced total distance of activity （P<0.05， P<0.01）， elevated OE% （P<0.05）， shortened sleep latency， and prolonged sleep time （P<0.05， P<0.01）. Compared with the serum MT in the blank control group， that in the model group was decreased （P<0.01）. Compared with the model group， no significant difference was observed in the low-dose CHC group， while other groups displayed increased serum MT （P<0.05）. The mRNA and protein expression of AANAT was decreased in the model group as compared with that in the blank control group （P<0.01）. Compared with the model group， the MT group and the high-dose CHC group showed up-regulated expression （P<0.05）.ConclusionCHC improved the behavioral indexes of PCPA-induced insomnia， increased the synthesis and secretion of MT in pineal cells， and elevated the serum MT level， which was related to the up-regulation of the mRNA and protein expression of AANAT in the pineal gland.Keywords：hypnotic active components of Cnidii Fructus;p-chlorophenylalanine （PCPA）-induced insomnia rats;melatonin synthesis rate-limiting enzyme arylalkylamine N-acetyltransferase （AANAT）- 19
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Mechanism of Artesunate in Reversing Epithelial-mesenchymal Transition and Akt /Snail Signaling Pathway in Colorectal Carcinoma
Abstract：ObjectiveTo investigate the effects of artesunate （ART） on epithelial-mesenchymal transformation （EMT） of colorectal cancer HCT-8 cells，and explore the effects of ART on cell migration，invasion，EMT ability， and protein kinase B （Akt）/Snail signaling pathway of colorectal cancer.Method3-（4-5-Dimethylthiazol-2-yl）2，5-diphenyltetrazolium bromide （MTT） assay was used to detect the effects of ART at different concentrations on the proliferation of HCT-8 cells. Wound healing assay and Transwell assay were used respectively to detect the effects of ART on migration and invasion of colorectal cancer cells. The effects of different concentrations of ART on the distribution of EMT-related proteins vimentin and E-cadherin in HCT-8 cells were detected by double-immunofluorescent staining. The effects of ART on protein expression levels of EMT markers E-cadherin，vimentin and N-cadherin in HCT-8 cells and the expression of Akt1， p-Akt1， and Snail1 in the Akt/Snail signaling pathway were determined by Western blot.ResultThe dose-dependent inhibitory effects of ART on the proliferation of HCT-8 cells were determined and the inhibition rate was calculated. A dose-response curve was plotted accordingly. The half-maximal inhibitory concentration （IC50） of ART on HCT-8 cells was （16.67±1.95） μmol·L-1. The following four groups were set up： a control group （0 μmol·L-1），and low-， medium-， and high-dose ART groups（2， 10， 50 μmol·L-1）. Compared with the results in the control group，ART inhibited the migration and invasion of HCT-8 cells（P<0.05）. Specifically， the expression of E-cadherin in HCT-8 cells was significantly up-regulated，and that of vimentin and N-cadherin was significantly down-regulated （P<0.05）. The expression levels of p-Akt1 and Snail1 were significantly decreased after ART treatment，thus inhibiting EMT（P<0.05）.ConclusionThe findings of this study suggested that ART inhibited the EMT-triggered migration and invasion of HCT-8 cells presumedly by inhibiting the activation of the Akt/Snail pathway to reverse EMT.Keywords：colorectal neoplasms;artesunate;epithelial-mesenchymal transition （EMT）;migration;invasion;protein kinase B （Akt）/Snail signaling pathway- 37
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Network Pharmacology-based Study of Bushen Zhuyun Prescription in Treatment of Recurrent Spontaneous Abortion
Abstract：ObjectiveTo explore the active components， targets， and signaling pathways responsible for Bushen Zhuyun prescription in treating the recurrent spontaneous abortion （RSA） based on network pharmacology and uncover its potential mechanism by molecular docking and in vitro cell experiments.MethodThe active components of Bushen Zhuyun prescription were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Traditional Chinese Medicine Integrated Database （TCMID） and the published articles， followed by the prediction of drug action targets based on such platforms as DrugBank and SwissTargetPrediction. GeneCards and Online Mendelian Inheritance in Man （OMIM） were searched to obtain the RSA targets， which were then intersected with the targets of Bushen Zhuyun Decoction. Following the plotting of Bushen Zhuyun prescription-compound-target-RSA network by Cytoscape 3.7.1， the protein-protein interaction （PPI） network was then constructed with STRING for screening the core network. The resulting common targets were then subjected to Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using R software. Autodock Vina 1.1.2 was used for molecular docking. The activation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/AKT） signaling pathway by Bushen Zhuyun prescription was verified in human umbilical vein endothelial cells （HUVEC） in vitro.ResultIt was found that 49 potential active components of Bushen Zhuyun prescription might act on 133 RSA targets. GO enrichment analysis yielded 470 biological processes， with angiogenesis， vascular development， cellular proliferation， and oxidative activity mainly involved. KEGG enrichment analysis revealed 103 signaling pathways （P<0.05）， and the PI3K/AKT signaling pathway， advanced glycation end product （AGE）/receptor for advanced glycation end product （RAGE） signaling pathway， and tumor necrosis factor （TNF） signaling pathway were the main ones. As indicated by molecular docking， the Vina scores of the main active component kaempferol with AKT1 and vascular endothelial growth factor A （VEGFA） were the lowest and similar. It was confirmed in vitro cell experiments that Bushen Zhuyun prescription activated the PI3K/AKT signaling pathway and up-regulated the expression of VEGFA and downstream AKT protein to promote angiogenesis.ConclusionBushen Zhuyun prescription promotes angiogenesis at the maternal-fetal interface by regulating angiogenesis and cellular proliferation， activating the PI3K/AKT pathway， and up-regulated the VEGFA expression， which is beneficial to the formation of placenta in early pregnancy and the maintenance of early pregnancy. This study has provided ideas for new drug development.Keywords：Bushen Zhuyun prescription;recurrent spontaneous abortion （RSA）;network pharmacology;mechanism of action;cell experiment- 15
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Regulatory Mechanism of Zedoary Turmeric Oil on VEGFA， STAT3 and mTOR in Ovarian Cancer
Abstract：ObjectiveTo explore the effects and mechanism of zedoary turmeric oil and its active components on the vascular endothelial growth factor A （VEGFA）， signal transducer and activator of transcription 3 （STAT3）， and mechanistic target of rapamycin （mTOR） in the ovarian cancer （OC）.MethodNetwork pharmacology technology was employed to analyze the mechanism of Curcumae Rhizoma on OC. Bioinformatics was used to analyze the expression of VEGFA， STAT3， and mTOR in OC and the effect on the prognosis of OC to explore the feasibility of zedoary turmeric oil in regulating VEGFA， STAT3， and mTOR in OC.The xenograft tumor model of nude mice was established， and the effects of zedoary turmeric oil and its active components on VEGFA， STAT3， and mTOR in OC were observed by hematoxylin-eosin （HE） staining， real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）， Western blot， and immunohistochemistry （IHC）.ResultBioinformatics analysis and literature research showed that VEGFA， STAT3， and mTOR played a special regulatory role in the occurrence and development of OC， and were potential key targets for the proliferation of OC. Network pharmacology analysis revealed that Curcumae Rhizoma could regulate multiple disease targets of OC， and mediate VEGFA， STAT3， and mTOR in OC through these multiple targets. As demonstrated by HE staining， the tumor cells in the model group were densely arranged， with no erosion on the edge and no vesicles inside. Compared with the model group， the cell density in other treatment groups was reduced， and strip-shaped erosion on the edge and small empty vesicles were observed in the tumor tissue， especially in the zedoary turmeric oil group. According to the results of Real-time PCR and IHC， zedoary turmeric oil and its active components could inhibit the mRNA and protein expression of VEGFA， STAT3， and mTOR in the OC tissue （P<0.05）.ConclusionZedoary turmeric oil and its active components could reduce the expression of VEGFA， STAT3， and mTOR in tumor tissue of nude mice， and inhibited the proliferation of OC through VEGFA， STAT3， and mTOR.Keywords：zedoary turmeric oil;ovarian cancer （OC）;curcumol;curgerenone;vascular endothelial growth factor A （VEGFA）;signal transducer and activator of transcription 3 （STAT3）;mechanistic target of rapamycin （mTOR）- 20
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Pharmacology
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Ganduqing Granules in Treatment of Common Cold Due to Qi Deficiency and Pathogen Invasion
Abstract：ObjectiveTo observe and evaluate the efficacy and safety of Ganduqing Granules in the treatment of common cold due to qi deficiency and pathogen invasion.MethodA multi-center， randomized， double-blind， and placebo-controlled clinical trial was conducted. One hundred and twenty patients were randomly divided into an experimental group （n=60） and a control group （n=60）. Patients in the experimental group were treated with oral Ganduqing Granules， 6 g per time， 3 times per day， while those in the control group received placebo， 6 g per time， 3 times per day. After treatment for 5 successive days in both groups， the overall remission rate， total score of primary symptoms， total score of secondary symptoms， total score of all symptoms， time required for overall symptom remission， and therapeutic efficacy against traditional Chinese medicine （TCM） syndrome in the two groups were analyzed in both full analysis set （FAS） and per-protocol set （PPS）， followed by the evaluation of safety in the safety set （SS）.ResultNo drop-out was found in the experimental group， whereas 3 cases in the control group dropped out. After 5 days of treatment， the overall remission rate of the experimental group was significantly higher than that of the control group （P<0.01）. The analysis in FAS and PDS revealed identical results. The experimental group was obviously better than the control group in improving the total scores of primary symptoms， secondary symptoms， and all symptoms （P<0.01）. The analysis results in FAS and PDS were consistent. There was no significant difference in overall remission time between the two groups. The experimental group was remarkably superior to the control group in alleviating such symptoms as aversion to wind and cold， nasal congestion， runny nose， fatigue， shortness of breath， laziness to speak， dry throat， sore throat， cough， and expectoration （P<0.05，P<0.01）. The analysis results in FAS were the same as those in PDS. All the patients did not present with significant abnormalities in vital signs， blood routine test， or liver and kidney function tests after medication. There was no significant difference in the incidence of adverse events between the experimental group and the control group.ConclusionGanduqing Granules effectively alleviate the symptoms of patients with common cold and shorten the course of disease， without inducing obvious side effects.Keywords：Ganduqing granules;common cold;syndrome of Qi deficiency and pathogen invasion- 22
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Effect of Modified Cangfu Daotantang on Metabolism and Pregnancy in Patients with Polycystic Ovary Syndrome
Abstract：ObjectiveTo observe the effect of modified Cangfu Daotantang on metabolism and pregnancy in patients with spleen deficiency and phlegm-dampness type polycystic ovary syndrome （PCOS）.MethodOne hundred and twelve patients were randomly divided into control group and observation group according to the random number table. Both groups took non-pharmacological interventions， oral metformin hydrochloride， 500mg/time， 3 times/day； oral ethinyl estradiol and cyproterone tablets， 1 tablet/time， 1 time/day， starting from the third to fifth day of menstruation and lasting for twenty-one days， for a total of 3 menstrual cycles. Patients in control group additionally took Erchen pills orally， 10 g/time， 2 times/day， while patients in observation group additionally took modified Cangfu Daotantang orally， 1 dose/day. The course of treatment was six menstrual cycles in both groups （or termination after conception）. The waist-to-hip ratio （WHR）， body mass index （BMI）， insulin resistance index （HOMA-IR）， pancreatic β-cell function （HOMA-β）， triglycerides （TG）， low-density lipoprotein （LDL） and non-high-density lipoprotein （nHDL） elevation after treatment were compared. The number of ovulation cycles monitored by B-ultrasound （6 menstrual cycles）， ovulation rate， human chorionic gonadotropin （HCG） day endometrial thickness， follicle diameter， cervical mucus score>8 points and endometrial morphology type A rate were measured and recorded. The recovery of menstruation， pregnancy and early miscarriage were recorded. Luteinizing hormone （LH）， estradiol （E2）， follicle stimulating hormone （FSH）， dehydroepiandrosterone sulfate （DHEAS）， testosterone （T）， anti-Müllerian hormone （AMH） levels， and insulin before and after treatment -Like growth factor-1 （IGF-1）， leptin （LP）， adiponectin （APN）， growth differentiation factor-9 （GDF-9） and tumor necrosis factor-α （TNF-α） levels were detected.ResultWHR， BMI and HOMA-IR levels of the observation group were lower than those of the control group （P<0.05， P<0.01）. HOMA-β level was higher than that in the control group （P<0.01）. The increase rates of LDL， TG， and nHDL in the observation group were 19.61%（10/51），25.49%（13/51），23.53%（12/51）， respectively， lower than 41.18%（21/51），47.06%（24/51），45.10%（23/51）respectively in the control group （χ2=5.607， χ2=5.131， χ2=5.263， P<0.05）. The menstrual recovery rate in the observation group was 90.20% （46/51）， higher than 72.55% （37/51） in the control group （χ2=5.239，P<0.05）. The observation group had more ovulation cycles than the control group （P<0.01）. The pregnancy rate in the observation group was 50.98% （26/51）， higher than 31.37% （16/51） in the control group （χ2=4.047，P<0.05）. On HCG day after treatment， the endometrial thickness and follicle diameter in the observation group were better than those in the control group （P<0.01）. The proportion of patients with cervical mucus score> 8 points was 78.43% （40/51） in the observation group， higher than 56.86% （29/51） in the control group （χ2=5.420，P<0.05）. The intimal morphology type A rate in the observation group was 52.94% （27/51）， higher than 31.37% （16/51） in the control group （χ2=4.864，P<0.05）. The levels of AMH， E2， DHEAS， LH， T ， IGF-1， LP and TNF-α in the observation group were lower than those in the control group （P<0.01）， while the APN and GDF-9 levels were superior to those in the control group （P<0.01）.ConclusionOn the basis of conventional western medicine intervention， modified Cangfu Daotantang can regulate abnormal metabolism and reproductive endocrine in patients with PCOS， improve conception， and regulate the expression of IGF-1， GDF-9， adipocytokines and inflammatory factors， improve ovulation and improve pregnancy rate.Keywords：polycystic ovary syndrome;infertility;spleen deficiency and phlegm dampness syndrome;Cangfu Daotantang;metabolic abnormalities;insulin-like growth factor-1;growth differentiation factor-9- 20
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Clinical Observation of Yishen Tongluo Decoction in Treating Chronic Glomerulonephritis with Spleen and Kidney Qi Deficiency and Blood Stasis Syndrome
Abstract：ObjectiveTo observe the clinical efficacy as well as anti-inflammatory and anti-fibrosis mechanism of Yishen Tongluo decoction in the treatment of chronic glomerulonephritis （CGN） with spleen and kidney Qi deficiency and blood stasis syndrome.MethodAccording to the random number table method， one hundred and twenty patients were divided into control group （60 cases） and observation group （60 cases）. The two groups were given oral valsartan capsules， 160 mg/time， 1 time/day， and dipyridamole tablets orally， 50 mg/time， 3 times/day. Patients in control group additionally took Wubi Shanyao pills orally， 9 g/time， 2 times/day， while patients in observation group additionally took Yishen Tongluo decoction orally， 1 dose/day. Both groups were treated continuously for four months. Before and after treatment， 24 h urine total protein （24 h UTP）， creatinine （SCr）， urea nitrogen （BUN）， serum cystatin C （CysC）， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， IL-17， interferon-γ （IFN-γ）， transforming growth factor-β1 （TGF-β1）， connective tissue growth factor （CTGF）， matrix metalloproteinase-9 （MMP-9）， tissue inhibitor of metalloproteinases （TMP-1） and hypoxia inducible factor-1α （HIF-1α） levels were detected in both groups. The scores of spleen and kidney Qi deficiency and blood stasis syndrome were graded. Urine protein， and urine red blood cells （urinary RBC） were monitored.ResultThe 24 h UTP， SCr， BUN and CysC levels of the observation group were lower than those of the control group （P<0.01）. The score of spleen and kidney Qi deficiency and blood stasis syndrome in the observation group was lower than that in the control group （P<0.01）. The levels of TNF-α， IL-6， IL-17 and IFN-γ in the observation group were lower than those in the control group （P<0.01）. The levels of TGF-β1， CTGF， TMP-1 and HIF-1α in the observation group were lower than those in the control group （P<0.01）. While MMP-9 level was higher than that in control group （P<0.01）. The clinical effective rate was 88.33% （53/60） in the observation group， higher than 73.33% （44/60） in the control group （- 29
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Postoperative Survival of Triple Negative Breast Cancer Patients After High Exposure to Traditional Chinese Medicine： A Cohort Study
Abstract：ObjectiveTo evaluate the 3-year survival outcomes of postoperative patients after high exposure to traditional Chinese medicine （TCM） for triple negative breast cancer （TNBC）.MethodThe complete 3-year follow-up data of 150 postoperative patients with stage I–III TNBC were retrospectively analyzed. All the patients received routine western medical treatments （surgery， chemotherapy， and/or radiotherapy） according to the National Comprehensive Cancer Network （NCCN） clinical practice guidelines in oncology as well as TCM. According to the degree of exposure to TCM， they were divided into the high- and low-exposure cohorts， with the oral administration of Chaihu Longmu Decoction with or without anti-cancer Chinese patent medicine for at least six months annually， or 18 months or more in the three years as the inclusion criterion for the former cohort. The metastatic sites of recurrent TNBC and the recurrent metastasis/death time were observed in both cohorts to compare the disease-free survival （DFS） and overall survival （OS）. The influences of onset age， pathological type， histopathological grade， vascular invasion， clinical stage， and exposure to TCM on survival were subjected to statistical analysis， followed by the observation of adverse effects.ResultThere was no significant difference in the metastatic sites between the two cohorts （P>0.05）. The high-exposure cohort had a longer 3-year DFS than the low-exposure cohort， and the 3-year DFS rate in the high-exposure cohort was increased by 16.9% （χ2=6.995， P=0.008） as compared with that in the low-exposure cohort， exhibiting a significant difference （P<0.05）. As revealed by the Cox proportional-hazards model， patients in the low-exposure cohort had a 3.724-fold as high risk of recurrent metastasis as that in the high-exposure cohort （95%CI 1.399~9.915）. There was no significant difference in the 3-year OS between the two cohorts （P>0.05）. The overall incidence of adverse effects in both groups was 7.3%， mainly manifested as gastrointestinal discomfort.ConclusionHigh exposure to TCM contributes to reducing postoperative recurrence and metastasis and prolonging DFS.Keywords：triple negative breast cancer （TNBC）;disease free survival （DFS）;retrospective cohort study;traditional Chinese medicine- 15
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Effect of Modified Wuzi Yanzongwan Periodic Staging Treatment on Outcome of Assisted Pregnancy in Patients with Poor Ovarian Response
Abstract：ObjectiveTo observe the effect of modified Wuzi Yanzongwan periodic staging treatment on the outcome of assisted pregnancy in patients with poor ovarian response （POR） and kidney deficiency syndrome.MethodOne hundred and four patients were randomly divided into observation group and control group， with 52 cases in each group. Both groups received gonadotropin releasing hormone （GnRH） antagonist regimen. The patients in control group additionally took Bushen Yutaiwan orally， 5 g/time， 3 times/day. The patients in observation group additionally took modified Wuzi Yanzongwan during pre-ovulation and post-ovulation periods， 1 dose/day. The treatment courses were 3 menstrual cycles （or termination after clinical pregnancy） in both groups. The number of eggs obtained， the number of available embryos， the number of fertilization， the number of high-quality embryos， the number of embryos implanted， the number of cycles cancelled， and the clinical pregnancy were recorded. Human chorionic gonadotropin （HCG） was injected， and then follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， estrogen （E2）， anti-Müllerian hormone （AMH）， basal antral follicle count （AFC） and endometrial thickness were measured daily. The number of days and dosage of Gn used， scores of kidney deficiency syndrome were recorded before and after treatment， and the adverse reactions during the study period were recorded.ResultThe number of eggs captured， rate of harvested eggs， number of available embryos， rate of available embryos， number of high-quality embryos， rate of high-quality embryos， and fertilization rate in observation group were higher than those in control group （P<0.05 or P<0.01）. The cycle cancellation rate was lower than that in the control group； the embryo implantation rate and clinical pregnancy rate were superior than those in control group， but the difference was not statistically significant. The FSH level and FSH/LH ratio in observation group were lower than those in control group during HCG day （P<0.01）， while E2， AMH， AFC and endometrial thickness were higher than those in control group （P<0.01）. Simultaneously， the number of days and amount of Gn used in observation group was lower than that in control group （P<0.01）.ConclusionThe Modified Wuzi Yanzongwan periodic staging treatment combined with GnRH antagonist scheme for patients with POR kidney deficiency syndrome， can regulate the level of endocrine hormones， promote follicular development， improve ovarian reserve， increase the number of eggs obtained， improve egg quality， help improve pregnancy outcomes， and increase the chances of successful pregnancy with assisted reproductive technology. It is worthy of further clinical research.Keywords：poor ovarian response;kidney deficiency syndrome;Wuzi Yanzongwan;ovarian reserve;periodic staging treatment;assisted reproduction- 23
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Clinic
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Mechanism of Astragali Radix Vesicle-like Nanoparticles for Reducing Blood Glucose in db/db Diabetic Mice by Regulating Gut Microbiota
Abstract：ObjectiveTo study the extraction method and characteristics of vesicle-like nanoparticles （VLNs） in Astragali Radix decoction， and to explore the mechanism of the VLNs in reducing blood glucose by regulating the gut microbiota of db/db diabetic mice.MethodUltracentrifugation and size exclusion chromatography were used to enrich VLNs from Astragali Radix decoction， and the morphology， particle size and concentration of the VLNs were analyzed by transmission electron microscope and nanoparticle tracking analyzer. The db/db diabetic mice were randomly divided into model group， Astragali Radix VLNs high-， medium- and low-dose （21.1， 10.6， 5.3 g·kg-1） groups and metformin group （0.25 g·kg-1） according to their blood glucose levels. There were 7 mice in each group， and another 7 C57BL/6 mice were set as the normal group. The mice were given intragastrically for 3 weeks （once a day）， and the changes of fasting blood glucose were observed every week. Hematoxylin-eosin （HE） staining was used to observe the pathological morphology of liver and pancreas of diabetic mice. The feces of mice were collected for 16S rRNA diversity detection of intestinal microbes.ResultThe size of the nanoparticles obtained by the two methods was about 200 nm. Astragali Radix VLNs extracted by ultracentrifugation had a typical saucer-like shape with the concentration of 3.0×1011 particles·mL-1. The morphology of Astragali Radix VLNs obtained by size exclusion chromatography was relatively poor with the concentration of 2.2×1011 particles·mL-1. After 3 weeks of administration， compared with the model group， Astragali Radix VLNs high-， medium- and low-dose groups could significantly reduce the fasting blood glucose of diabetic mice （P<0.05， P<0.01）. The VLNs could improve the gut microbiota dysbiosis， significantly decrease the ratio of Firmicutes/Bacteroidota， and increase the relative abundance of beneficial bacteria and reduce the relative abundance of harmful bacteria.ConclusionAstragali Radix VLNs may reduce the blood glucose of db/db diabetic mice by adjusting the ratio of Firmicutes/Bacteroidota in the intestinal flora.Keywords：Astragali Radix;vesicle-like nanoparticles;ultracentrifugation;size exclusion chromatography;fasting blood glucose;intestinal flora;particle size distribution- 21
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Investigation of Potential Pharmacodynamic Substances and Mechanism of Gardeniae Fructus in Treatment of Ischemic Stroke Based on HPLC-Q-TOF-MS/MS and Network Pharmacology
Abstract：ObjectiveCombined with high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry （HPLC-Q-TOF-MS/MS） and network pharmacology， to predict the target and potential mechanism of Gardeniae Fructus in the treatment of cerebral ischemia.MethodHPLC-Q-TOF-MS/MS was used to identify the chemical constituents of Gardeniae Fructus according to the retention time， relative molecular weight， secondary mass spectrometry fragmentation and other information of chromatographic peaks， and combined with literature data. The targets of main active ingredients in Gardeniae Fructus were predicted by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and SwissTargetPrediction database. The potential targets of Gardeniae Fructus against cerebral ischemia were obtained through Online Mendelian Inheritance in Man （OMIM）， GeneCards and Kyoto Encyclopedia of Genes and Genomes （KEGG）. Gene ontology （GO） function enrichment and KEGG pathway analysis of potential targets were analyzed with the DAVID 6.8. Cytoscape 3.6.0 software was used to construct the network of active components-targets-pathways. At last， Discovery Studio 2016 software was applied in the molecular docking verification between the key active ingredients and potential protein targets.ResultA total of 40 chemical constituents in Gardeniae Fructus were identified， including iridoids， diterpenoid pigments， organic acids， monoterpenoids and other components. According to the main active ingredients， 208 potential targets were predicted， 560 disease targets related to cerebral ischemia were retrieved， 59 key targets were selected by mapping component targets with disease targets. These targets could act on key target proteins such as tumor necrosis factor （TNF）， Caspase-3 （CASP3） and CASP8， and participate in the regulation of TNF， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， hypoxia inducible factor 1 （HIF-1） and other signal pathways. Molecular docking results showed that geniposide could interact with targets of prostaglandin G/H synthase 2 （PTGS2）， TNF and nuclear transcription factor-κB p65 （RELA）， crocin Ⅰ could interact with interleukin-2 （IL-2）.ConclusionGeniposide， crocin Ⅰ and other ingredients in Gardeniae Fructus can play a role of anti-inflammatory and inhibiting apoptosis to prevent or treat cerebral ischemic diseases by up-regulating protein expression level of RELA and IL-2， down-regulating protein expression level of TNF， CASP8， CASP3 and matrix metalloproteinase 2 （MMP2）， and regulating TNF， PI3K/Akt and HIF-1 signaling pathways.Keywords：Gardeniae Fructus;cerebral ischemia;network pharmacology;molecular docking;inflammatory cytokines;signaling pathways- 33
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Effects of Exogenous Substances on Growth of
Polyporus umbellatus Mycelium and Its Polysaccharide ContentAbstract：ObjectiveTo explore the effects of diverse exogenous substances at different concentrations on the growth of Polyporus umbellatus mycelium and polysaccharide content and screen out the optimal growth condition for P. umbellatus mycelium， so as to provide a reference for its large-scale artificial cultivation.MethodP. umbellatus mycelium was cultured in media containing different exogenous substances using the method for fungal culturing in plate. The growth rate of the mycelium was judged by the colony diameter and the polysaccharide content was determined by the phenol-sulfuric acid method.ResultThe high-dose cyclic adenosine monophosphate， 6-benzyl aminopurine （6-BA）， gibberellic acid （GA）， 2，4-dichlorophenoxyacetic acid （2，4-D）， vitamin （V） B1， VB3， VB6， VB9， and VB12 all promoted the growth of P. umbellatus mycelium and elevated polysaccharides content. By contrast， indole acetic acid （IAA）， VC， and VB2 inhibited its growth， with the most obvious inhibition detected in the high-dose VC group. IAA and VB2 both reduced the polysaccharide content， whereas the high-dose VC significantly increased the polysaccharide content. Cyclic adenosine monophosphate， 6-BA， GA， 2，4-D， VB1， VB3， VB6， VB9， and VB12 at the concentrations of 2 mmol·L-1， 6 mg·L-1， 15 mg·L-1， 2 mg·L-1， 4 mg·L-1， 2 mg·L-1， 4 mg·L-1， 6 mg·L-1， and 10 mg·L-1， respectively， contributed to the growth of P. umbellatus mycelium and polysaccharide accumulation.ConclusionThe growth of P. umbellatus mycelium and polysaccharide accumulation can be regulated by adding exogenous substances to the culture medium.Keywords：exogenous substances;Polyporus umbellatus mycelium;growth rate;polysaccharide content- 16
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Analysis of Equivalence Between Traditional and Nontraditional Medicinal Varieties of Citri Reticulatae Semen from Sichuan Province
Abstract：ObjectiveTo study the differences in genetic relationship， shape， size， and flavonoid content between traditional and nontraditional medicinal varieties of Citri Reticulatae Semen produced in Sichuan province as well as their equivalence.MethodSix batches of traditional medicinal Citri Reticulatae Semen （Citrus reticulata 'Dahongpao'） and 23 batches of nontraditional medicinal varieties were collected， and their genetic relationship was explored using sequence-related amplified polymorphism （SRAP） markers. Following the observation of their shapes and sizes under a stereomicroscope， the contents of naringin， hesperidin， and neohesperidin were measured by high performance liquid chromatography （HPLC）. SIMCA 14.1 software was used for cluster analysis of their shapes， sizes， and flavonoid contents， thus figuring out the similarities between the traditional and nontraditional medicinal varieties in character， size， and chemical components.ResultSRAP markers-based genetic relationship analysis effectively distinguished different Citri Reticulatae Semen varieties from each other. Some samples collected from the same or adjacent places exhibited a close genetic relationship and they shared high similarities in shape， size， and flavonoid content. However， the traditional medicinal Citri Reticulatae Semen was still quite different from most nontraditional medicinal varieties.ConclusionThe analysis of differences in genetic materials， appearance， character， and active ingredient content between the traditional and nontraditional medicinal varieties revealed that the equivalence of C. reticulata 'Ponkan' samples from some regions with the traditional medicinal variety was the largest， enabling them to be considered as the emerging medicinal variety.Keywords：Citri Reticulatae Semen;traditional medicinal variety;nontraditional medicinal varieties;sequence-related amplified polymorphism （SRAP）;characters;flavonoids- 15
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Simulation Analysis of Occupancy Rates of Baicalein， Quercetin and Galangin on Target Sites of Xanthine Oxidase
Abstract：ObjectiveTo simulate the occupancy rates of baicalein， quercetin and galangin on the target sites of xanthine oxidase in vivo.MethodIn this experiment， the half inhibitory concentration （IC50） of febuxostat， baicalein， quercetin and galangin against xanthine oxidase were determined by in vitro enzymatic reaction. Binding free energy was predicted by molecular docking technology and their association rate constant （kon） and dissociation rate constant （koff） were determined by surface plasmon resonance technology. Based on measured binding kinetic parameters （kon and koff） and extracted pharmacokinetic data， the target occupancy model in vivo was established.ResultThe IC50 values of febuxostat， baicalein， quercetin and galangin were 0.002 7， 1.63， 0.38， 1.59 µmol·L-1， respectively. The IC50 of febuxostat was very close to that reported in the literature. The predicted curve of target occupancy rate in vivo of febuxostat was consistent with its duration of clinical efficacy. When single intragastric administration of long-circulating liposomes of quercetin with dose of 100 mg·kg-1 in rats， the time of target occupancy rate >70% in vivo lasted for about 3.9 h. When rats were orally administered baicalein and galangin with dose of 200 mg·kg-1， the time of target occupancy rate >50% in vivo lasted for about 10 h and 1.7 h， respectively.ConclusionThe prediction model of xanthine oxidase target occupancy constructed by drug target binding kinetics and in vivo pharmacokinetic curves can effectively evaluate the in vivo inhibitory activity of compounds against the target.Keywords：xanthine oxidase;target occupancy;drug target binding kinetics;association rate constant （kon）;dissociation rate constant （koff）;half inhibitory concentration （IC50）;surface plasmon resonance （SPR）- 20
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Analysis of Material Basis and Mechanism of Sangjiang Ganmao Injection in Treatment of Common Cold Based on UPLC-Q-Orbitrap HRMS and Network Pharmacology
Abstract：ObjectiveTo explore the material basis and mechanism of Sangjiang Ganmao injection （SG） in the treatment of common cold by ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） and network pharmacology.MethodUPLC-Q-Orbitrap HRMS was used to identify the chemical components of SG with mobile phase of acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-10 min， 4%-15%A； 10-35 min， 15%-30%A； 35-45 min， 30%-33%A； 45-55 min， 33%-60%A； 55-58 min， 60%A）， flow rate of 0.2 mL·min-1， electrospray ionization （ESI） and scanning range of m/z 100-1 500 under positive and negative ion modes. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and GeneCards 5.0 database were used to screen and predict the potential targets of chemical components in SG， STRING 11.0 database and Cytoscape 3.7.2 software were used to construct protein-protein interaction （PPI） network model， gene ontology （GO） analysis and pathway analysis were performed on potential targets by Metascape 3.5， Reactome database and Kyoto Encyclopedia of Genes and Genomes （KEGG）， Cytoscape 3.7.2 software was used to build the network of "herbs-ingredients-key targets".ResultA total of 54 components in SG were identified， and 80 potential targets of SG for treatment of common cold were predicted and screened based on this. SG exerted therapeutic effects by acting on targets such as interleukin （IL）-6， tumor necrosis factor （TNF） and IL-10， and signaling pathways such as IL-17 signaling pathway， TNF signaling pathway and interaction of cytokine receptors.ConclusionSG may interfere with the expression of inflammatory cytokines by acting on related targets and pathways such as inflammation and immune system， and regulate the immune function of the body as a whole， thereby exerting a therapeutic effect.Keywords：Sangjiang Ganmao injection;Sangjuyin;common cold;network pharmacology;material basis;inflammation;immune system- 19
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Soil Physical and Chemical Properties， Microorganisms and Metabolites in Different Culture Environments of
Gastrodia elata Abstract：ObjectiveTo study the soil physical and chemical properties， microorganisms， and metabolites in different culture environments of Gastrodia elata， so as to provide scientific basis for subsequent cultivation of G. elata in multiple environments.MethodThe tubersphere soil of G. elata cultured in different environments was collected for analyzing the soil nutrients， microbial numbers， and metabolite differences using the agrochemical method， plate-count method， and gas chromatography-time-of-flight mass spectrometry （GC-TOF-MS）-based non-targeted metabonomic approach.ResultThe analysis of soil physical and chemical properties revealed the highest soil moisture， pH， available potassium， and available phosphorus in the spinney and the highest electrical conductivity， total nitrogen， alkali-hydrolyzable nitrogen， and organic matter in the pinewood. As demonstrated by the quantitative analysis of soil microorganisms， the cultivable microorganisms were bacteria， actinomycetes， and fungi， with the bacterial population and total microbial biomass in the spinney and the number of fungi and actinomycetes in the barren slope detected to be the largest. The ratio of bacteria to fungi （B/F value） in the pinewood was the highest， while that in the barren slope was the lowest. The results of metabonomic research demonstrated that the compositions and quantities of soil metabolites in the spinney （Z group）， pinewood （S group）， and barren slope （HD group） varied. Through comparisons between S and Z groups， between HD and Z groups， as well as between HD and S groups by orthogonal partial least squares discriminant analysis （OPLS-DA）， 18， 35， and 24 differential metabolites were separately screened out， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis yielded 5， 9， and 13 metabolic pathways. There existed a significant causal relationship of the soil physical and chemical properties and microbial numbers with the metabolites.ConclusionThe soil physical and chemical properties， microbial numbers， and metabolite changes differed significantly in different culture environments of G. elata， which were sorted by the suitability in a descending order as follows： spinney > pinewood >barren slope. The soil physical and chemical properties and microbial numbers are the crucial factors driving changes in soil metabolites， suggesting that regulating the soil physical and chemical characteristics and microbial characteristics in the culture environment is an important mechanism for maintaining the G. elata-soil-microbial symbiotic system.Keywords：Gastrodia elata;culture environment;soil physical and chemical properties;soil microbial numbers;soil metabolites- 17
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Pharmacy Fundamentals
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Syndrome Distribution of 9 323 Cough Cases in Ancient Chinese Medical Books Based on Latent Structure Model and Logistic Regression Analysis
Abstract：ObjectiveTo explore the syndrome distribution of cough cases in ancient Chinese medical books.MethodA total of 9 323 cough cases in the database of lung diseases in ancient Chinese medical books were extracted. Lantern 5.0 was used to construct the latent structure model for the 100 manifest variables based on the first 50 symptoms and 50 Chinese herbal medicines， and different latent nodes were interpreted. The syndrome differentiation patterns of syndromes with symptoms and Chinese herbal medicine （formula） were quantitatively revealed by the comprehensive clustering weights of latent structure. The correlation of diseases with syndromes was fitted through the binary Logistic regression analysis.ResultThere were 204 syndromes involved in 9 323 cough cases with 18 syndromes showing a frequency higher than 100. As demonstrated by the model established on the first 50 symptoms and 50 Chinese herbal medicines， 35 latent variables， 98 latent classes， and 10 comprehensive clustering models were obtained， where Z5 was the highest in the threshold value （6.7）， Z6 in the information coverage of productive cough （52%）， and Z7 in the score （19）. The binary Logistic regression model fitted the correlation between different disease types and five syndromes， where the dominance ratio of external wind to the syndrome of wind-heat invading lung reaching up to 88.919， those of syndrome of phlegm-heat accumulating in lung to diseased heat and sputum 51.594 and 15.861， and those of the syndrome of phlegm-dampness obstructing lung to dampness， phlegm， and fluid retention 31.415， 34.370， and 4.936， respectively.ConclusionThe newly discovered cough syndromes included lung heat and yin deficiency， external cold and internal fluid retention， weakness of spleen and stomach， and phlegm and blood stasis in lung. In most cases， multiple syndromes were observed clinically， such as syndrome of deficiency of both Qi and Yin in lung combined with yin deficiency in lung and kidney. Since differentiation of traditional Chinese medicine （TCM） syndrome is subjective， the weight can indicate the difference in the contributions of different symptoms to the syndrome， which is of guiding significance for syndrome inference. The latent structure model combined with Logistic regression analysis can solve the problem of quantification in TCM syndrome differentiation and can be used to explore the syndrome distribution of diseases.Keywords：latent structure model;Logistic regression analysis;cough;syndrome- 15
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Exploration of Potential Mechanisms of Corylin Against Lung Cancer By High-throughput Sequencing
Abstract：ObjectiveTo explore the potential molecular mechanism of corylin in the treatment of lung cancer.MethodA549 cells were treated with different concentrations of corylin， and their proliferation was detected using methye thiazolye telrazlium （MTT） reagent. Then the trend analysis and gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were conducted to screen the key genes and pathways of corylin against A549 cell proliferation， followed by the verification of sequencing results by real-time polymerase chain reaction （Real-time PCR）.ResultCorylin inhibited the proliferation of A549 cells and regulated the expression of 4 364 genes in cells. The trend analysis revealed that these genes were clustered into 20 distinct modules， among which four were significantly down-regulated， suggesting that corylin exerted the anti-proliferation effect by inhibiting the expression of some genes. The inter-group comparison of differentially expressed genes （DEGs） showed that the elevation in the concentration of corylin resulted in more down-regulated genes but weakened proliferation， consistent with the findings by trend analysis. The GO and KEGG enrichment analysis of 278 DEGs in the high-dose corylin group demonstrated that corylin mainly changed the cellular and metabolic processes， which was attributed to its regulation of steroid biosynthesis， fatty acid metabolism， biosynthesis of unsaturated fatty acids， synthesis and degradation of ketone bodies， and steroid hormone biosynthesis. The Real-time PCR results confirmed that corylin down-regulated the mRNA expression levels of LSS， stearoyl-CoA desaturase （SCD）， 3-hydroxy-3-methylglutaryl-CoA synthase 1 （HMGCS1）， but up-regulated the mRNA expression of recombinant human angiopoietin-like protein 4 （ANGPTL4）， basically consistent with the transcriptomics results.ConclusionCorylin inhibits A549 cell proliferation and alleviates lung cancer by targeting the related genes in lipid metabolism pathways.Keywords：corylin;A549;lipid metabolism;high-throughput sequencing;cancer- 26
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Construction of Tibetan Medicine Prescription Database for Treating "Gnyan-rims" and Analysis of Their Medication Law Based on Python Language
Abstract：ObjectiveTo construct the database of Tibetan medicine prescriptions for "Gnyan-rims" disease， and to explore the invisible medication law of Tibetan medicine in the treatment of "Gnyan-rims" disease， such as prescription compatibility and combination of drug properties.MethodThe prescriptions for treating "Gnyan-rims" were retrieved from four Tibetan medical literatures such as The Four Medical Tantras， Kong-sprul-zin-tig， Phyag-rdor-gso-rig-phyogs-bsgrigs and Sman-sbyor-lag-len-phyogs-bsgrigs， and the database was constructed under Python code， and the Apriori algorithm and the vector structure model of taste property flavor transformation were used for analysis.ResultAccording to the characteristics of Tibetan medicine prescription data， with six fields of prescription name， formula， dosage， efficacy， source and original text as the core， a Tibetan medicine treatment "Gnyan-rims" prescription database with functions of cleaning， searching and exporting was established. A total of 7 602 prescriptions were included in the database， among which 598 prescriptions had therapeutic effects of "Gnyan" and "Rims". The results of compatibility analysis showed that Shexiang， Hezi， Honghua， Mukuer Moyao， Tiebangchui， Tianzhuhuang and Bangga were the most frequently used drugs， while the correlation degrees of Shexiang-Mukuer Moyao， Honghua-Tianzhuhuang， Shexiang-Hezi and Shexiang-Tiebangchui were the strongest， and all the drug composition of Wuwei Shexiang pills appeared in the top ten correlations. According to the property analysis of 40 prescriptions containing high-frequency drugs， 19 prescriptions were found to have excessive bitter taste， followed by 9 prescriptions such as Sanchen powders with excessive sweetish taste， and the ratios of sweetish and bitter tastes in six tastes were >35%. The total of sweetish and bitter prescriptions accounted for 70% of the total prescriptions. Among the three flavors， the bitter flavor was the most abundant. The cool effect， dull effect and heavy effect were prominent among the seventeen effects.ConclusionThe prescription database of Tibetan medicine for "Gnyan-rims" can promote the high-quality development of research on prevention and treatment of plague with ethnic medicine. Tibetan medicine treatment of "Gnyan-rims" focuses on the composition of Wuwei Shexiang pills， with the property combination of "cool-bitter and sweet-bitter flavor-cool， dull and heavy"， which mainly treats diseases such as "heat sharp light-mkhris pa-heat". These studies can provide data basis and theoretical reference for the selection of Tibetan medicine prescription and its composition for treating plague.Keywords：Tibetan medicine;plague;formulas;coronavirus disease 2019 （COVID-19）;database;Ro-nus-zhu-rjes;Python code- 28
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Mechanism of Resveratrol in Treatment of Lung Adenocarcinoma： An Exploration Based on Database Mining and Molecular Biology
Abstract：ObjectiveTo explore the mechanism of resveratrol （RSV） in the treatment of lung adenocarcinoma （LUAD） based on bioinformatics and molecular biology.MethodThe targets of RSV were retrieved from DrugBank and then imported into STRING for constructing a protein-protein interaction （PPI） network.TCGA database was utilized to analyze the expression of target genes in tumor and normal tissues， followed by the prediction of their impacts on tumor occurrence and development and the screening of target genes using random forest and univariate Cox regression models.With the results of bioinformatics taken into consideration， the mechanism of RSV in inhibiting LUAD was further explored by molecular biology.ResultTen Hub genes were screened out from the PPI network of RSV targets.Among them， solute carrier family 2 member 1 （SLC2A1）， arachidonate 5-lipoxygenase （ALOX5）， peroxisome proliferative activated receptor gamma （PPARG）， and arachidonate 15-lipoxygenase （ALOX15） differed significantly in their expression in tumor and normal tissues.As revealed by random forest and univariate COX regression analysis， SLC2A1 was of great significance to the survival and prognosis of patients with LUAD.The survival analysis through Kaplan-Meier （KM） plotter indicated that the SLC2A1 expression was closely related to the overall survival （OS）， first progression （FP）， and post-progression survival （PPS） of LUAD patients.The molecular biological experiments further proved that RSV inhibited the proliferation and migration of LUAD cells by reducing the expression of SLC2A1.As verified by immunohistochemical scoring， SLC2A1 protein expression in tumor tissue was significantly different from that in normal tissue.ConclusionRSV inhibits the proliferation and migration of LUAD cells by reducing the expression of SLC2A1， which has far-reaching significance in the clinical treatment of LUAD.Keywords：resveratrol （RSV）;lung adenocarcinoma （LUAD）;bioinformatics;mechanism of action;molecular biology- 32
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Data Mining
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Expert Consensus on Otorhinolaryngology Head and Neck Diseases Responding Specifically to Traditional Chinese Medicine
Abstract：Traditional Chinese medicine （TCM） and western medicine have their respective advantages and limitations in the diagnosis and treatment of common otorhinolaryngology head and neck diseases. Although the integrated TCM and western medicine exhibits definite curative effects， there is no consensus on the otorhinolaryngology head and neck diseases responding specifically to TCM or integrated TCM and western medicine， as well as the diagnosis and treatment schemes. The China Association of Chinese Medicine （CACM） thus organized the otorhinolaryngology head and neck specialists of both TCM and western medicine to discuss the etiology， pathogenesis， and clinical diagnosis and treatment methods of common otorhinolaryngology head and neck diseases with the results of multiple clinical trials taken into account. The acute pharyngitis， chronic pharyngolaryngitis， paraesthesia pharyngis， hysterical aphasia， allergic rhinitis， subjective tinnitus， and otogenic vertigo were confirmed to respond specifically to TCM or integrated TCM and western medicine. Then a mutually agreed diagnosis and treatment scheme and recommendation with integrated TCM and western medicine was formulated as a reference for clinical practice， thus benefiting more patients.Keywords：otorhinolaryngology head and neck surgery;integrated traditional Chinese medicine （TCM） and western medicine;diseases responding specifically to TCM;expert consensus- 31
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Dominant Disease
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Treatment of Thin Endometrium in Infertility Based on Thought of "Blood Being Basis of Women"
Abstract：Infertility is commonly seen but difficult to treat in the department of obstetrics and gynecology. The incidence of infertility in women of childbearing age has reached as high as 25% in China， which seriously affects social harmony and people's happiness. As the lifestyle of modern women changes， thin endometrium has gradually become an important reason for infertility and embryo transfer failure. Such factors as repeated operations in uterine cavity and oral intake of hormones have resulted in destructed blood vessels and reduced blood flow in endometrium， impaired growth of glandular epithelial cells， insufficient local blood supply， retarded development of endometrium， reduced receptivity， and therefore poor acceptance of endometrium to the implanted gestational sac， which is not conducive to the implantation of fertilized eggs. As a result， the infertility will occur. Traditional Chinese medicine （TCM） has a long history of treating infertility. During the long-term clinical practice， physicians of ancient and modern times have gained rich experience in the diagnosis and treatment of infertility. The idea of "blood being the basis of women" sprouted in the Spring and Autumn and Warring States Periods and determined in the Song Dynasty as an important guiding principle that can be applied in various physiological periods of women. It emphasizes "blood regulation" for gynecological diseases to ensure sufficient blood in blood vessels and its normal circulation， thus allowing the thoroughfare and conception vessels and endometrium to be nourished and the pregnancy to be facilitated. With blood as the basis， the paper explored the physiopathological correlations of five zang organs and thoroughfare and conception vessels with blood， pointed out the preference for "blood harmonization" in ancient literature based on the differentiation of infertility into blood stasis， blood cold， blood heat， and blood deficiency syndromes， reviewed the key methods of modern physicians in assisting pregnancy including tonifying kidney and nourishing blood， tonifying kidney and activating blood， tonifying kidney and invigorating spleen， as well as invigorating kidney and soothing liver， and summarized the efficacy of integrated TCM and western medicine in promoting ovulation， regulating hormones， improving blood supply， and nourishing endometrium to assist pregnancy. All these would contribute to the clinical treatment and scientific research of thin endometrium in infertility from multiple angles.Keywords：blood being the basis of women;thin endometrium in infertility;five Zang organs;thoroughfare and conception vessels- 25
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Guarantee of Quality and Clinical Efficacy of Chinese Materia Medica Preparation： Control of Quality Transfer Process of Chinese Materia Medica
Abstract：Traditional Chinese medicine （TCM） is the treasure of the Chinese nation. As an important raw material for clinical treatment of diseases， Chinese materia medica plays an extremely important role. However， in the process of transformation from traditional wild collection of animals and plants to modern artificial cultivation and industrial production of preparations， whether the quality of Chinese materia medica is fully transferred will directly affect the quality and clinical efficacy of Chinese materia medica preparation. From the field to the sickbed， process control of quality transfer of Chinese materia medica is the key to guarantee quality and curative effect. In this paper， the whole process that affects the quality of Chinese materia medica preparations such as seed and seedling， planting and breeding， harvesting and processing， processing of decoction pieces and preparation production was analyzed. Paying attention to the whole process of quality control of Chinese materia medica is of great significance to improve the quality of Chinese materia medica preparations and promote the rapid development of TCM. Based on this， the author intended to analyze the key control links in the quality transfer process of Chinese materia medica （breeding， planting areas and field management， timely harvesting and intensive primary processing， appropriate processing， optimization of preparation technology， standardization of packaging and informationization of storage and transportation）， in order to provide reference for the design and development of Chinese materia medica preparations guided by clinical value.Keywords：Chinese materia medica;quality transfer;germplasm resources;pollution-free planting;harvesting and processing;processing of decoction pieces;preparations;quality evaluation- 18
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Academic Discussion
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Tripterygium wilfordii in Treatment of Kidney Diseases and Its Toxic and Side Effect： A ReviewAbstract：In recent years， with the rapid development of traditional Chinese medicine industry in China， the efficacy of Chinese medicinals in treating disease and maintaining health has been increasingly recognized. Tripterygium wilfordii， a Chinese medicinal for expelling wind， dredging collaterals， removing dampness， and relieving pain， is commonly used for treating acute and chronic glomerulonephritis， rheumatoid arthritis， and other diseases. However， the frequent occurrence of adverse reactions has limited its wide application in clinical practice. The existing studies have gradually confirmed that T. wilfordii and its active ingredients exert the bidirectional effects on kidney function. This paper reviewed the related clinical applications and articles published in the past decades and summarized the material basis for its bidirectional effects and the specific action mechanisms in renal protection and renal damage. It was found that the main active ingredients in T. wilfordii were tripterygium glycosides and triptolide， which exerted the protective or toxic and side effects on kidney by regulating immunity， influencing mitogen-activated protein kinases （MAPK） pathway， and changing the expression and function of renal transporters. Besides， the roles of administration time， dosage， and body status in the exertion of protective or toxic and side effects by T. wilfordii were also discussed. The review aimed to provide new ideas for the research on the treatment of kidney diseases with T. wilfordii and its safety application.Keywords：Tripterygium wilfordii;renal protection;renal damage;bidirectional effects;chemical ingredients- 24
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Efficacy of Spleen-invigorating Chinese Medicinals in Repairing Intestinal Mucosal Barrier Injury： A Review
Abstract：As the first line of defense between the intestinal environment and the outside world， the intestinal mucosal barrier is essential for maintaining the intestinal homeostasis. The intestinal mucosal barrier injury will change the intestinal permeability and allow bacterial translocation and the entry of endotoxins into blood， thus triggering a series of inflammatory responses， followed by the injury of related tissues and the aggravation of primary diseases. The spleen， the acquired foundation， is responsible for maintaining the internal and external balance of the body and resisting external evils. Its physiological function is similar to that of the intestinal mucosal barrier. Spleen deficiency easily leads to intestinal mucosal barrier dysfunction. Therefore， replenishing Qi， invigorating spleen， and restoring the efficacy of spleen and stomach qi in defensing and governing transportation and transformation are the keys to prevent and treat intestinal mucosal barrier injury. In recent years， studies have shown that the spleen-invigorating Chinese medicinals repair the intestinal mucosal injury by promoting the expression of intestinal epithelial tight junction proteins， regulating the intestinal immune function， microbial flora， and metabolites， and supplementing the intestinal nutrition， enabling them to gradually become a research hotspot. After reviewing the relevant articles published in China and abroad， this paper expounded the common syndrome types of traditional Chinese medicine （TCM）， the changes in intestinal mucosal barrier induced by spleen deficiency， the repairing effects of spleen-invigorating Chinese medicinals on intestinal mucosal barrier injury， in order to provide some clues for the research on the treatment of intestinal mucosal barrier dysfunction-related diseases with spleen-invigorating Chinese medicinals.Keywords：invigorating spleen;intestinal mucosal barrier;Chinese medicinals;injury;research progress- 25
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25 -
Pathogenesis of Vitiligo and Its Prevention and Treatment with Traditional Chinese Medicine： A Review
Abstract：Vitiligo is a common pigmentary disorder characterized by localized or generalized depigmentation of the skin， resulting in milky white or light pink patches with smooth surface but no rashes.Modern medicine believes that it is closely related to immune function， oxidative stress， autoimmune destruction of melanocytes， heredity， and neurochemical factors， but the specific pathogenesis is still unclear.Due to the easy diagnosis but hard management and the high recurrence rate， vitiligo has seriously affected the quality of life and mental health of patients.At present， it is mainly treated with glucocorticoids， immunomodulators， vitamin D3 derivatives， and antioxidants in western medicine， and the resulting short-term outcomes are satisfactory.However， a series of side effects may be caused by the long-term use.Traditional Chinese medicine （TCM） has a long history of dealing with vitiligo.Guided by the rich experience accumulated in daily practice， it exerts the preventive and therapeutic effects against vitiligo via multiple components， multiple targets， and multiple pathways， without inducing obvious adverse reactions， which has made it widely concerned by many doctors and scholars.As the research concerning the prevention and treatment of vitiligo with TCM intensifies， more and more single Chinese medicinals， Chinese medicinal monomers， and compound prescriptions have been proved to play a therapeutic role via multiple mechanisms.After reviewing the articles on the alleviation of vitiligo with TCM published in the past five years and retrieved from the literature databases， this paper summarized the efficacy of TCM in regulating immune dysfunction， improving oxidative stress injury， protecting melanocyte function， adjusting mitochondrial structure and function， and controlling the alteration of intestinal micro-flora and abnormal microRNA （miRNA） expression， so as to clarify the pathogenesis of vitiligo and provide theoretical and scientific basis for the in-depth study and clinical application of TCM in the prevention and treatment of vitiligo.Keywords：vitiligo;pathogenesis;traditional Chinese medicine（TCM）;research progress- 34
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<HTML><CITATION> <Bulk Citation>Published：2021-06-25
Review
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