Aims To establish the methods of primary culture of fibroblast-like synoviocytes in rats with adju-vant arthritis(AA-FLS)and analyze the feature and to investigate the possibility of AA-FLS as the model for the RA in vitro.Methods The synovial cells obtained from the SD rats were immunized by the Mtb and iden-tified by morphology and immunocytochemistry.The viability of AA-FLS was assessed by Cell Counting Kit-8.ELISA was applied to detect TNF-αand IL-1 βin cell media. Apoptosis was measured by Hochest 33258.The expressions of mitochondrial apoptosis-re-lated molecules,including Bcl-2,Bax,Pro-caspase-3 and Cleave-caspase-3 were determined by Western Blot.Result In isolated primary synovial cells,more
than 95% of AA-FLSs were fusiform.Immunocyto-chemistry result showed a positive expression of vimen-tin and a negative expression of CD68 in AA-FLS.Cell proliferation of AA-FLS was higher than FLS and cell apoptosis of AA-FLS was curbed.Western blot data demonstrated that the protein expressions of Bcl-2,Bax were regulated and the expression of caspase-3 was ac-tivated in AA-FLS.Conclusions AA-FLS is biologi-cally characterized by high level proliferation activity and inflammatory cytokines and apoptosis suppression. AA-FLS can be used as the model for the RA in vitro.

OBJECTIVETo characterize the proteomic profiles of joint synovial tissue in normal rats and rats with rheumatoid arthritis (RA) and identify the proteins related with the occurrence of RA to explore the pathogenesis of RA.

METHODSSD rat models of RA were established using Mtb (heat-killed Mycobacterium tuberculosis H37Ra). Two-dimensional gel electrophoresis proteomics technology was employed to analyze the difference in synovial tissue protein profiles between RA model rats and normal rats, and two of the differentially expressed proteins were verified with Western blotting and fluorescence quantitative PCR.

RESULTSComparison of the two-dimensional gel electrophoresis patterns from the model rats and normal rats showed 4 up-regulated and 4 down-regulated proteins by 2 folds in the RA model rats. Western blotting and fluorescent quantitative PCR of 2 of the 8 proteins yielded consistent results with those by proteomics analysis.

CONCLUSIONArthritis synovial lesions in RA represent very complex pathological processes involving a variety of proteins, and these differentially expressed proteins may contribute to the progression of RA.

Animals ; Arthritis, Rheumatoid ; metabolism ; Blotting, Western ; Disease Models, Animal ; Disease Progression ; Electrophoresis, Gel, Two-Dimensional ; Polymerase Chain Reaction ; Proteins ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; metabolism ; pathology ; Up-Regulation

OBJECTIVE:To study the improvement effects of Rouganbao granules on liver fibrosis of model rats,and to provide experimental evidence for further development of this preparation. METHODS:Totally 45 male SD rats were divided into normal group,model group,positive control group(Fuzheng huayu capsules,0.4 g/kg)and Rouganbao granules high-dose and low-dose groups(16.8,8.4 g/kg by crude drug)according to random number table,with 9 rats in each group. Except for normal group,other groups were given CCl4sub cutaneously and high lipid diet+15% ethanol solution for 12 weeks to establish liver fibrosis model. After modeling,medication groups were given relevant medicine intragastrically,once a day,for consecutive 12 weeks. Normal group and model group were given corresponding volume of water intragastrically. The pathology change of liver tissues of rats was observed by inverted microscope. The levels of liver function indexes (ALT,AST,ALB) and liver fibrosis indexes(HA,Ⅳ-C,LN,Ⅲ-PC)were determined by ELISA. RESULTS:Compared with model group,liver cell vacuolation and liver fibrosis of rats were improved significantly in Rouganbao granule high-dose and low-dose groups;central vein was clearly visible,and no dilation and atrophy were found,especially in high-dose group;the serum levels of ALT and AST were decreased significantly,while the level of ALB was increased significantly(P＜0.05 or P＜0.01),of which the protective effect for liver was similar to positive control drug but better in increasing ALB(P＜0.05 or P＜0.01);the serum levels of HA,Ⅳ-C,LN and Ⅲ-PC were all decreased in varying degree,especially in high-dose group(P＜0.05 or P＜0.01),of which the effect of relieving liver fibrosis was similar to positive control drug. CONCLUSIONS:Rouganbao granules can improve liver function and delay the progression of liver fibrosis in model rats.

Chrysoeriol is a natural flavonoid compound, which is widely present in many kinds of traditional Chinese medicine and medicinal herbs. In recent years, studies of Chrysoeriol on the pharmacological effects and its glycosides have gradually increased, especially in anti-tumor, anti-oxidative damage and anti-inflammatory immune regulation, etc., showing good pharmacological effects, and it has prospective potency as a candidate to develop new drug in those domain. This article briefly reviews the pharmacological effects and underlying mechanisms of chrysoeriol, so that researchers can understand the pharmacological characteristics of this compound, and also provide references for the development of new drugs based on this ingredient.

Objective: To investigate the role of exosome (EXO) transporting Let-7a to regulate MYC gene in the malignant biological behaviors of triple negative breast cancer (TNBC) cell, and to explore the underlying mechanism. Methods: After the completion of cell culture, the gene and protein expressions of MYC and Let-7a in TNBC MDA-MB-231cells were detected by qPCR and WB, respectively. Recombinant lenti-virus vector carrying Let-7a and Crisper/Cas-9 system with MYC knockdown were transfected into MDA-MB-231 cells; MTT assay, Transwell assay and Scratch healing assay were performed to examine the proliferation, invasion and migration of MDA-MB-231 cells. Luciferase activity assay was performed to validate the binding between MYC and Let-7a. EXO was isolated and identified by transmission electron microscopy and WB assay in wild-type and Let-7a over-expressed MDA-MB-231 cells, respectively. After co-incubation of two types of EXO and MDA-MB-231 cells, the effects of Let-7a on biological behaviors of MDAMB-231 cells via EXO were detected by qPCR, WB, MTT and Transwell etc. Results: Let-7a was negatively correlated with MYC in breast cancer tissues and cell lines (all P<0.05); MYC promoted while Let-7a inhibited the proliferation, migration and invasion of breast cancer cells (all P<0.01); Let-7a silenced MYC by acting on 3'UTR of MYC gene, thereby reducing the expression of MYC protein (P<0.05); Let-7a was enveloped by EXO and transported to cancer cells, there by inhibiting the proliferation, migration and invasion of MDA-MB-231 cells. Conclusion: EXO some mediated Let-7a silences MYC gene by acting on its 3'UTR region, thus inhibiting the proliferation, migration and invasion of MDA-MB-231 cells.