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Rheumatoid arthritis (RA) is a common disease worldwide and is treated commonly with methotrexate (MTX). CS12192 is a novel JAK3 inhibitor discovered by Chipscreen Biosciences for the treatment of autoimmune diseases. In the present study, we examined the therapeutic effect of CS12192 against RA and explored if the combinational therapy of CS12192 and MTX produced a synergistic effect against RA in rat collagen-induced arthritis (CIA). Arthritis was induced in male Sprague-Dawley rats by two intradermal injections of bovine type II collagen (CII) and treated with MTX, CS12192, or the combination of CS12192 and MTX daily for two weeks. Effects of different treatments on arthritis score, X-ray score, pathology, and expression of inflammatory cytokines and biomarkers were examined. We found that treatment with either CS12192 or MTX produced a comparable therapeutic effect on CIA including: (1) significantly lowering the arthritis score, X-ray score, serum levels of rheumatic factor (RF), C-reactive protein (CRP), and anti-nuclear antibodies (ANA); (2) largely alleviating histopathological damage, reducing infiltration of Th17 cells while promoting Treg cells; (3) inhibiting the expression of inflammatory cytokines and chemokines such as IL-1β, TNF-α, IL-6, CCL2, and CXCL1. All these inhibitory effects were further improved by the combinational therapy with MTX and CS12192. Of importance, the combinational treatment also resulted in a marked switching of the Th17 to Treg and the M1 to M2 immune responses in synovial tissues of CIA. Thus, when compared to the monotherapy, the combination treatment with CS12192 and MTX produces a better therapeutic effect against CIA with a greater suppressive effect on T cells and macrophage-mediated joint inflammation.

Additional file 1: Supplementary methods: 1) Hong Kong Diabetes Register TRS Study Group Members; 2) Hong Kong Diabetes Biobank Study Group Members; and 3) Cohort descriptions. Figure S1. Principal component analysis (PCA). Figure S2. Pooled correlations of each candidate polygenic risk scores with measured lipid traits. Figure S3. Proportion of phenotypic variance in lipid traits explained by each candidate polygenic risk scores. Figure S4. Geometric means of measured lipid traits stratified by the quintile of polygenic risk score with the best performance. Figure S5. Geometric means of measured lipid traits at baseline and follow-up, and three-year changes in lipid traits stratified by quintile of polygenic risk scores in adolescents.

Background: Natural small molecules have become more attractive as alternatives to non-steroidal anti-inflammatory drugs in osteoarthritis (OA) treatments. This study aims to investigate the effects of Asiatic acid (AA) on OA in chondrocytes and the surgery-induced OA animal model. Methods: Cytotoxicity of AA in primary rat articular chondrocytes was determined. Chondrocytes were pretreated with AA at the safe concentrations and subsequently treated with IL-1β. The production of inflammatory mediators including nitric oxide (NO), nitric oxide synthase (iNOS), as well as cyclooxygenase (COX)-2, and the expression of chondrogenic and hypertrophic markers including Sox 9, Aggrecan, Col 2a1, and matrix metalloproteinase-13 (MMP13) in the cells were measured. The effect of AA on nuclear factor-kappa B (NF-κB) signaling pathway was further determined by dual luciferase assay and western blot. The surgery-induced OA animals were treated with AA or saline for 6 weeks. The pathological changes in the affected joints were measured by micro-CT and histological analysis. Results: We found a broad safety spectrum of AA from 0 to 25 μM. A dose-dependent inhibitory effect of AA on NO production, as well as iNOS and COX-2 expression were found. Meanwhile, AA promoted chondrogenesis and inhibited hypertrophy in chondrocyte treated with IL-1β. In addition, AA inhibited NF-κB signaling pathway with a dose-dependent manner. Furthermore, results from animal study revealed that AA prevented articular cartilage damage as well as subchondral bone remodeling in the surgery-induced OA animal.

Additional file 2: Fig. S1. Full Western blots used for Fig. 1b, c, d, e. (a) Full Western blots used for Fig. 1b. (b) Full Western blots used for Fig. 1c. (c) Full Western blots used for Fig. 1d. (d) Full Western blots used for Fig. 1e. Fig. S2. Full Western blots used for Fig. 2b, d, e, f, g, h, i, j. (a) Full Western blots used for Fig. 2b. (b) Full Western blots used for Fig. 2d. (c) Full Western blots used for Fig. 2e. (d) Full Western blots used for Fig. 2f. (e) Full Western blots used for Fig. 2g. (f) Full Western blots used for Fig. 2h. (g) Full Western blots used for Fig. 2i. (h) Full Western blots used for Fig. 2j.Fig. S3. Full Western blots used for Fig. 3a, b, c, d, e, f, h, j, k. (a) Full Western blots used for Fig. 3a. (b) Full Western blots used for Fig. 3b. (c) Full Western blots used for Fig. 3c. (d) Full Western blots used for Fig. 3d. (e) Full Western blots used for Fig. 3e. (f) Full Western blots used for Fig. 3f. (g) Full Western blots used for Fig. 3h. (h) Full Western blots used for Fig. 3j. (i) Full Western blots used for Fig. 3k. Fig. S4. Full Western blots used for Fig. 4a, b, c, d, e, f, g. (a) Full Western blots used for Fig. 4a. (b) Full Western blots used for Fig. 4b. (c) Full Western blots used for Fig. 4c. (d) Full Western blots used for Fig. 4d. (e) Full Western blots used for Fig. 4e. (f) Full Western blots used for Fig. 4f. (g) Full Western blots used for Fig. 4g. Fig. S5. Full Western blots used for Fig. 5a, b, c, d, e, f. (a) Full Western blots used for Fig. 5a. (b) Full Western blots used for Fig. 5b. (c) Full Western blots used for Fig. 5c. (d) Full Western blots used for Fig. 5d. (e) Full Western blots used for Fig. 5e. (f) Full Western blots used for Fig. 5f. Fig. S6. Full Western blots used for Fig. 6a, b, c, d, e, f, g, h, j, k, l, m, n, o, q. (a) Full Western blots used for Fig. 6a. (b) Full Western blots used for Fig. 6b. (c) Full Western blots used for Fig. 6c. (d) Full Western blots used for Fig. 6d. (e) Full Western blots used for Fig. 6e. (f) Full Western blots used for Fig. 6f. (g) Full Western blots used for Fig. 6g. (h) Full Western blots used for Fig. 6h. (i) Full Western blots used for Fig. 6j. (j) Full Western blots used for Fig. 6k. (k) Full Western blots used for Fig. 6l. (l) Full Western blots used for Fig. 6m. (m) Full Western blots used for Fig. 6n. (n) Full Western blots used for Fig. 6o. (o) Full Western blots used for Fig. 6q. Fig. S7. Full Western blots used for Fig. 8a, b, c, d, e. (a) Full Western blots used for Fig. 8a. (b) Full Western blots used for Fig. 8b. (c) Full Western blots used for Fig. 8c. (d) Full Western blots used for Fig. 8d. (e) Full Western blots used for Fig. 8e. Fig. S8. Full Western blots used for Fig. 9b, d, f, g, j. (a) Full Western blots used for Fig. 9b. (b) Full Western blots used for Fig. 9d. (c) Full Western blots used for Fig. 9f. (d) Full Western blots used for Fig. 9g. (e) Full Western blots used for Fig. 9j.

Background & Aims: The role of macrophages in fibrosing steatohepatitis is largely unclear. We characterized the origin and molecular mechanisms of macrophages and its targeted therapy in fibrosing steatohepatitis. Methods: Fibrosing steatohepatitis was established in Alms1 mutant (foz/foz) and C57BL/6 wildtype mice fed high-fat/high-cholesterol (HFHC) or methionine-and choline-deficient (MCD) diet. Bone marrow transplantation (BMT) was performed to track the macrophage origin in fibrosing steatohepatitis. Macrophages were depleted by liposomal clodronate. Primary macrophages were isolated from bone marrow for adoptive transfer into mice. Results: Macrophage infiltration is significantly induced in two mouse models of fibrosing steatohepatitis and human NASH-fibrosis patients. Bone marrow-derived macrophages (BMMs) contribute to the hepatic macrophage accumulation in experimental fibrosing steatohepatitis. Hepatic BMMs depletion by liposomal clodronate significantly attenuated fibrosing steatohepatitis. This effect was associated with reduced hepatic stellate cell (HSC) activation, collagen deposition and hepatic expression of key pro-fibrotic factors (TIMP1, TIMP2 and TGFβ1) and endoplasmic reticulum (ER) stress markers (GRP78, IRE1α and PDI). Conversely, adoptive transfer of BMMs significantly aggravated fibrosing steatohepatitis. Moreover, macrophage conditioned medium directly promoted the phenotypic transition of primary quiescent HSCs to activated HSCs; enhanced activation and proliferation (P

Disease associated gene deficient embryonic stem cells can serve as valuable in vitro models to study disease mechanisms and screen drugs. Smad3 mediated TGF-β/Activin/Nodal signaling plays important roles in many biological processes. Despite numerous studies regarding Smad3 function, the role of Smad3 in mouse ES cells is not well studied. To understand the function of Smad3 in mouse ES cells, we derived Smad3−/− ES cells and wild type ES cells. Smad3−/− ES cells display no defect on self-renewal. They express similar level of pluripotent genes and lineage genes compared to wild type ES cells. However, Smad3 ablation results in transient difference in germ layer marker expression during embryoid body formation. Mesoderm lineage marker expression is significantly reduced in the embryoid body formed by Smad3−/− ES cells compared to wild type ES cells. Intriguingly, subcutaneous injection of Smad3−/− ES cells into nude mice leads to formation of malignant immature teratomas, whilst wild type ES cells tend to form mature teratomas. Smad3−/− ES cell formed teratomas can therefore provide a new model for the study of the mechanism of malignant teratomas.