Objective To explore the effects of umbilical cord-derived mesenchymal stem cells (UCMSCs) on fibroblast-like synoviocytes(FLSs) from patients with rheumatoid arthritis(RA). Methods collagen-induced arthritis(CIA) models were developed on Wistar rats and 1 ×106 UCMSCs were given by intravenous injection from tail vein on the 17th day. On day 42, rats were sacrificed and synovial tissues were obtained to detect matrix metalloproteinase (MMP)-1, MMP-3 and MMP-13. Synovial tissues from patients with RA and osteoarthritis(OA) treated by knee arthroplasty were used to isolate FLSs. RNA of FLSs were extracted to compare MMP-1, MMP-3 and MMP-13 expression. FLSs and UCMSCs were cocultured through transwell for 72 hours. Then levels of MMPs were compared in the supernatants by Luminex. The MMPs expressed by FLSs and soluble factors expressed by MSCs were detected by real time-polymerase chain reaction(PCR). After adding antibodies to soluble factors, the MMPs expressions of RA FLSs were compared. Results MMP-1 (6.9±5.4, 1.3±1.4, P<0.05), MMP-3 (6.0±6.5, 1.4±1.0, P<0.05) and MMP-13 (21.8± 20.8, 1.5±1.6, P<0.05) expression were much higher in CIA rats compared with healthy controls. MMP-1 (1.3±1.4, 6.9±5.4,8.7±6.8, P<0.05), MMP-3(1.4±1.5, 6.0±6.5, 6.0±5.7, P<0.05) and MMP-13(3.0±3.2,
22±21, 22±26, P<0.05) expression were inhibited by UCMSCs in vivo. In vitro, MMP-1 (1.8±0.9, 0.9±0.7, t=2.44, P<0.05), MMP-3(2.6±1.7, 1.1±1.0, t=2.25, P<0.05) and MMP-13(2.4±2.3, 0.6±0.7, t=2.37, P<0.05) levels were higher in RA than OA FLSs. After coculture, MMP-13(1.3±1.2, 0.9±1.2, t=3.63, P<0.05) expressed by FLSs were down-regulated, however MMP-1 (1.5±1.4, 6.6±6.0, t=3.90, P<0.05) and MMP-3 (7±17, 22±35, t=2.86, P<0.05) were up-regulated when analyzed by paired t-test. Soluble factors such as I-DO, HGF and IL-10 were elevated. Anti-IL-10 antibody could decrease the function of UCMSCs by inhibiting MMP-13 expression in RA FLSs. Conclusion UCMSCs ameliorates RA by secreting soluble factor IL-10, which may inhibit MMPs expressed by FLSs.

Objective To measure the level of interleukin (IL)-27 in sera of patients with Sj(o)gren's syndrome (SS) and investigate its role in IL-10 in CD3+CD8-T cells.Methods Serum levels of 1L-27 and IL-10 from 34 SS patients and 33 healthy controls were tested by enzyme linked immunosorbent assay (ELISA).Peripheral blood monocytes (PBMCs) from SS patients were collected and cultured in different conditions (one with PHA,the other with PHA and IL-27).Seventy-two hours later,cells were harvested and the percentages of CD3 +CD8-IL-10+ cells were measured by flow cytometry.Expressions of c-Maf,IL-10 transcription factor,were examined using real-time polymerase chain reaction (PCR).The data were analyzed by t test,Pearson's correlation analysis and multiple linear regression analysis.Results Serum level of IL-27 in patients with SS [(158±34) pg/ml] was significantly higher than that in healthy controls [(139±18) pg/ml,t=2.823,P＜0.01].IL-27 level was positively correlated with IL-10 and IgG levels (r=0.384,P＜0.05; r=0.354,P＜0.05),but negatively correlated with SSDAI (r=0.503,P＜0.01).Multiple regression analysis showed independent correlation of IL-27 with IL-10 and SSDAI in SS patients (β=0.412,P＜0.01; β=-0.525,P＜0.01),but not IgG.CD3+CD8-IL-10+ cells was upregulated by IL-27 in vitro (P＜0.01,n=14),so was c-Maf (P＜0.01,n=22).Conclusion IL-27 could ameliorate disease activity of Sjogren's syndrome.

Objective:To observe the anti-inflammatory and glycometabolic effects of glutathione(GSH)on macrophages in collagen induced arthritis(CIA)mice.Methods:①CIA model establishment and groups:A total of 14 female DBA/1J mice were ran-domly divided into:CIA+PBS group and CIA+GSH group.The mice were sacrificed on the 50th day,collecting serum and isolating bone marrow derived macrophages(BMDM),which were marked as BMDM1.②Trained immunity model establishment and groups:BMDM were isolated from normal DBA/1J mice,and were pretreated with histone H3K27 demethylases inhibitor(GSKJ1)or PBS for 2 h.Then,serum from CIA model mice in vivo was incubated for 24 h,and the samples were grouped as follows:(CIA+GSH)+PBS group,(CIA+GSH)+GSKJ1 group,(CIA+PBS)+PBS group,(CIA+PBS)+GSKJ1 group.Lipopolysaccharide(LPS)was adopted to stimulated cells on the 6th day,which were marked as BMDM2.③RNA sequencing was used to detect differentially expressed genes(DEGs)and their function in BMDM1 and BMDM2.q-PCR was adopted to estimate the mRNA levels of PFK and Idh3g.The culture supernatants were used to measure the protein levels of TNF-α and IL-6 by ELISA.Results:①Compared with CIA+PBS group,the mice in CIA+GSH group showed lighter of joint swelling(P<0.05),less arthritis index(P<0.05),HE staining suggested less inflam-matory cell infiltration,Safranin O-fast green staining showed more chondrocytes,TRAP staining indicated reduced osteoclasts.②In BMDM1,GO analysis showed that DEGs were mainly involved in glutathione derivative metabolic process,IL-6 production,inflammatory response,innate immune response,regulation of primary metabolic process and glycolipid binding,compared with CIA+GSH and CIA+PBS group.In CIA+GSH group,the mRNA level of PFK was significantly decreased(P<0.05),while Idh3g was also significantly up-regulated(P<0.05),and the expression of TNF-α,IL-6 were both reduced(P<0.05)compared with CIA+PBS group.③In BMDM2,GO analysis showed that DEGs were also involved in inflammatory response,activation of innate immune response,regulation of tumor necrosis factor production,positive regulation of IL-6 production,regulation of glycolytic process and 1,3-β-D-glucan binding between CIA+GSH and CIA+PBS group.Furthermore,in(CIA+GSH)+PBS group,PFK was decreased(P<0.05),Idh3g was up-regulated(P<0.05),and IL-6 was also significantly down-regulated(P<0.05)compared with(CIA+PBS)+PBS group.However,there was no significant difference in the expression of Idh3g and PFK,moreover,TNF-α and IL-6 were significantly up-regulated compared with(CIA+GSH)+GSKJ1 group and(CIA+GSH)+PBS group.Conclusion:GSH can regulate glycometabolism and inflammatory response of macrophages via demethylation of histone H3K27,and it can also alleviate CIA in mice.