It is proposed that the pathogenesis of rheumatoid arthritis （RA） is centered on deficiency， dampness， and blood stasis， which interact with and evolve into one another during the onset and progression of the disease. The development of RA is closely associated with insufficiency of healthy qi and the interbinding of dampness and blood stasis. Accordingly， treatment emphasizes an integrated approach that combines tonifying deficiency， eliminating dampness， and resolving blood stasis， and is implemented in three main stages. In the initial stage， therapy focuses on supporting healthy qi， dispelling dampness， and relieving impediment， with modified Huangqi Guizhi Wuwu Decoction （黄芪桂枝五物汤） combined with Yiyiren Decoction （薏苡仁汤）. In the active stage， treatment aims to eliminate dampness， resolve blood stasis， and unblock the collaterals， using modified Wutou Decoction （乌头汤） or Guizhi Shaoyao Zhimu Decoction （桂枝芍药知母汤）. In the remission stage， therapy emphasizes strengthening the spleen and reple-nishing qi to prevent recurrence， with modified Shenling Baizhu Powder （参苓白术散） combined with Guipi Decoction （归脾汤）.

Objective:To investigate whether autophagy in the inflammatory environment of thrombotic antiphospholipid syndrome (APS) drives polymorphonuclear neutrophils (PMNs) to release neutrophil extracellular traps (NETs) and whether IFN-λ1(IL-29) has a function of inhibiting NETs-related thrombin production.Methods:PMNs isolated from peripheral blood of healthy volunteers were in vitro stimulated by serum samples of patients with active APS. IL-29 and 3-methyladenine (3-MA) were used to regulate NETs release and autophagy. NETs expression was evaluated by immunofluorescence technique (IFT) and flow cytometry (FCM). Autophagy-related proteins were detected by Western blot. The levels of thrombin-antithrombin (TAT) complexes were determined by enzyme-linked immunosorbent assay (ELISA). Results:The percentages of NETs released from PMNs of healthy volunteers after stimulation with serum samples of APS patients were (22.8±3.1)% according to IFT and (10.1±2.7)% according to FCM and used as control group. IL-29 and 3-MA inhibited the release of NETs from PMNs stimulated by serum samples of APS patients and the percentages of NETs were (5.3±2.2)% and (4.9±2.4)% detected by IFT, and (2.1±1.3)% and (2.7±1.4)% by FCM, respectively. There were significant differences with the control group ( q=9.89, 10.67, 11.74, 9.61, all P<0.01). IL-29 inhibited the expression of LC3B, but promoted the expression of p62 on PMNs of healthy volunteers after stimulating with serum samples of APS patients, and the differences with the group without IL-29 pretreatment were statistically significant [LC3B/GAPDH: (0.28±0.03)% vs (0.77±0.06)%, q=8.65; p62/GAPDH: (0.63±0.05)% vs (0.33±0.03)%, q=4.78; both P<0.01]. IL-29 and 3-MA reduced the levels of NETs-related TAT complexes from (7.6±0.6) ng/ml to (3.1±0.5) ng/ml and (4.7±0.4) ng/ml, respectively ( q=6.34 and 5.15, both P<0.01). Conclusions:Autophagy was involved in the formation of NETs and subsequent coagulation cascade activation in PMNs of healthy subjects after stimulation with serum samples of APS patients with thrombosis. IL-29 suppressed the production of NETs and NETs-associated thrombin by inhibiting autophagy.

PURPOSE: Cancer-associated fibroblasts (CAFs) activated by cancer cells has a central role in development and malignant biological behavior in colorectal cancer (CRC). Adult fibroblasts do not express Snail, but Snail-positive fibroblasts are discovered in the stroma of malignant CRC and reported to be the key role to chemoresistance. However, the reciprocal effect of CAFs expressed Snail to chemoresistance on CRC cells and the underlying molecular mechanisms are not fully characterized. MATERIALS AND METHODS: Snail-overexpressed 3T3 stable cell lines were generated by lipidosome and CT26 mixed with 3T3-Snail subcutaneous transplanted CRC models were established by subcutaneous injection. Cell Counting Kit-8, flow cytometry and western blotting assays were performed, and immunohistochemistry staining was studied. The cytokines participated in chemoresistance was validated with reverse transcriptase-polymerase chain reaction and heatmap. RESULTS: Snail-expression fibroblasts are discovered in human and mouse spontaneous CRCs. Overexpression of Snail induces 3T3 fibroblasts transdifferentiation to CAFs. CT26 co-cultured with 3T3-Snail resisted the impairment from 5-fluorouracil and paclitaxel in vitro. The subcutaneous transplanted tumor models included 3T3-Snail cells develop without restrictions even after treating with 5-fluorouracil or paclitaxel. Moreover, these chemoresistant processes may be mediated by CCL1 secreted by Snail-expression fibroblasts via transforming growth factor β/nuclear factor-κB signaling pathways. CONCLUSION: Taken together, Snail-expressing 3T3 fibroblasts display CAFs properties that support 5-fluorouracil and paclitaxel chemoresistance in CRC via participation of CCL1 and suggest that inhibition of the Snail-expression fibroblasts in tumor may be a useful strategy to limit chemoresistance.
Adult ; Animals ; Blotting, Western ; Cell Count ; Cell Line ; Colorectal Neoplasms* ; Cytokines ; Drug Resistance, Multiple ; Fibroblasts* ; Flow Cytometry ; Fluorouracil ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Injections, Subcutaneous ; Mice ; Paclitaxel ; Snails ; Transforming Growth Factors

Objective To analyze hemolysin and virulence -related genes in incomplete hemolytic Staphylococcus aureus.Methods Fifty strains of incomplete hemolytic Staphylococcus aureus were isolated from patients admitted in the Second Affiliated Hospital of Soochow University during 2013 and 2014, and the isolates with complete hemolytic phenotype were also collected at the same period as the control strains . All the strains were inoculated and subcultured on four kinds of sheep blood agar plates supplied by different manufacturers to compare their hemolytic phenotype .The relative mRNA expressions of hemolysin genes (hla, hlb, hlc, hld) in standard strain, complete and incomplete hemolytic phenotype strains were detected by real-time quantitative polymerase chain reaction (RT-qPCR), and valued by 2 -△△Ct method.t test was used to compare mRNA expressions of hemolysin genes .Western blot was performed to analyze the expression of α-hemolysin.Antibiotic susceptibility test of incomplete hemolytic strains was performed using broth microdilution method.Resistant gene mecA and virulence genes pvl, tst were detected by PCR.Results The steady and hereditary incomplete hemolysis was observed in 50 strains of incomplete hemolytic Staphylococcus aureus on the sheep blood agar plates from different suppliers .Taking mRNA expression of hla, hlb, hlc, hld in standard strain as 1, the relative mRNA expressions of hemolysin genes in incomplete hemolytic strains were 0.02, 7.51, 0.06 and 0.12 respectively, there were statistical differences between standard strain and incomplete hemolytic strains (t =8.46, -56.40, 8.12 and 7.61, all P <0.05).And the expression of α-hemolysin was decreased in incomplete hemolytic strains .All the strains were identified as methicillin resistant Staphylococcus aureus (MRSA).Three strains exhibited different minimum inhibitory concentrations of teicoplanin and linezolid after subcultured , but the differences had no impact on the final results of antibiotic susceptibility test .mecA, pvl and tst genes were positive in incomplete hemolytic strains . Conclusion Staphylococcus aureus with incomplete hemolytic phenotype is methicillin resistant with higher expression of β-hemolysin and lower expressions of α-hemolysin, γ-hemolysin and δ-hemolysin.It carries plv and tst virulence genes and is of high virulence .