Aim To investigate the estrogenic activities of four components from pregnant mare’s urine extract. Methods The estrogenic activities of four components were assessed using two in vitro tests:the MCF-7 cell proliferation assay (E-screen test)and the luciferase transfected CHO cell gene reporter assay.In the lucifer-ase reporter gene assay,the reporter gene plasmids PGM-ERE-Luc and ERαor ERβand a control plasmid (pRL-cmv)were transiently co-transfected into CHO cells to establish an ERα-or ERβ-cell screening system which was used to measure estrogenic activity of four compounds.Results MCF-7 cells treated with HP, DHP,PT and HA significantly proliferated,thereby of-fering in vitro evidence for the estrogenic activities of HP,DHP,PT and HA,and they showed dose-depend-ent activities.Compared EC50 of PE and RPE with that of E2 ,HP,DHP,PT and HA exerted relatively weak estrogenic activities.The in vitro ER-mediated reporter gene assay revealed that HP,DHP,PT and HA dis-played estrogenic activities mediated by ERβor ERα. Compared with the EC50 of E2 ,HP,DHP,PT and HA exhibited lower estrogenic potencies.Conclusion HP, DHP,PT and HA possess weaker estrogenic activities than E2 .

Teaching practice is an important link in the process of postgraduate training. This paper analyzes the current situation and existing problems of postgraduate teaching practice, and discusses the influence of new teaching practice mode on postgraduate training. Aiming at the problems existing in the teaching practice of postgraduate students, this paper carries out the reform from five aspects: subject course learning, teaching preparation, theoretical teaching, experimental teaching participation and teaching management participation, and evaluates the implementation effect of the reform. Through the reform of postgraduate teaching practice, the teaching ability and social competitiveness of postgraduate students have been effectively improved. The new teaching practice mode provides new ideas and methods for the cultivation of postgraduate students' teaching ability.

ObjectiveTo preliminarily predict the targets and signaling pathways of indole-3-methanol in the treatment of obesity based on molecular docking technology and network pharmacology， and then verify the prediction results by the experiment in vitro. MethodThe pharmacological targets of indole-3-methanol were obtained from SwissTargetPrediction and literature review. Obesity-related targets were obtained from Online Mendelian Inheritance in Man （OMIM）， GeneCards， and Comparative Toxicogenomics Database （CTD）. The protein-protein interaction network of the targets of indole-3-methanol and obesity was built by STRING. Cytoscape 3.8.2 was used for target screening. Gene ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment were performed for the common targets shared by obesity and indole-3-methanol in DAVID 6.8. AutoDock Vina 1.1.2 was employed to perform the molecular docking between indole-3-methanol and disease targets. Finally， the in vitro experiment was carried out to verify the anti-obesity effect of indole-3-methanol. ResultIndole-3-methanol and obesity shared 80 common targets， which included matrix metalloproteinase （MMP）-9， Janus kinase （JAK） 2， etc. KEGG enrichment predicted that indole-3-methanol mainly acted on tumor necrosis factor （TNF）， vascular endothelial growth factor （VEGF）， tyrosine kinase receptor 2 （ErbB2）， and epidermal growth factor receptor （EGFR） signaling pathways in the treatment of obesity. Molecular docking showed that indole-3-methanol had good binding activity with fat mass and obesity-associated protein （FTO）. The results of Western blot， MTT assay， and oil-red O staining showed that indole-3-methanol down-regulated the expression of FTO in 3T3-L1 cells （P<0.05）. ConclusionIndole-3-methanol may treat obesity by down-regulating the expression of FTO protein and further inhibiting adipocyte proliferation. This study provides an experimental basis for deciphering the anti-obesity mechanism of indole-3-methanol.

Objective To predict the potential mechanism of Punicalagin in the treatment of Inflammatory bowel disease by network pharmacology.Methods The intersection genes of Punicalagin and IBD were obtained from the database,and PPI,GO and KEGG pathways were enriched and analyzed.Punicalagin and the target were verified by molecular docking.C57BL/6J mice were drunk dextran sulfate sodium to establish inflammatory enteritis model,and were given Punicalagin for 7 d of intervention.During the administration,signs of mice in each group were observed,daily disease activity index was calculated;Intestinal permeability test after administration;The colon tissue was stained with hematoxylin eosin to observe the pathological changes and calculate the histological damage score;Detection of tumor necrosis factor(TNF-α),Interleukin-10(IL-10),myeloperoxidase(MPO),chemokine 1(CXCL1)and other cytokines in colon tissue of mice by ELISA.Detection of TNF-α,IL-6,MPO and CXCL1 level in mouse serum by ELISA.CCK8 method was used to determine the effect of Punicalagin on the proliferation activity of caco-2 cells.The levels of cytokines released by caco-2 cells induced by lipopolysaccharide(LPS)were detected by ELISA.Results 14 common targets of Punicalagin and IBD were obtained,including tumor necrosis factor(TNF),arachidonic acid-5-lipoxygenase(ALOX5)and vascular endothelial growth factor A(VEGFA).KEGG enrichment analysis predicted that the treatment of IBD by Punicalagin mainly acted on arachidonic acid signaling pathway,age-rage signaling pathway,VEGR signaling pathway and Ras signaling pathway.Molecular docking showed that Punicalagin had good docking activity with TNF receptor.Compared with the model group,the decreasing range of body mass in Punicalagin group abated(P<0.01);the disease activity index of Punicalagin group decreased significantly(P<0.01);The congestion and edema of colonic mucosa were significantly reduced,and the histological injury score was significantly reduced(P<0.01);The level of TNF-α,IL-1β,MPO,CXCL1,IL-6,IL-18,IFN-γ in colon tissue was significantly decreased(P<0.01);20-300 μmol·L-1 Punicalagin promoted caco-2 cell proliferation and inhibited TNF-α secretion induced by LPS,up-regulation of IL-10 levels.Conclusion Punicalagin inhibits the secretion of TNF-α and other proinflammatory factors,up-regulation of the level of anti-inflammatory factor IL-10,and improvement of colonic inflammatory response in IBD mice.