Dynamic expression and significance of Bcl6 in fibroblast-like synoviocytes induced osteoclast differentiation and activation in rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the dynamic expression and significance of B cell lymphoma (Bcl) 6 in fibroblast-like synoviocytes (FLS) induced osteoclast differentiation and activation in rheumatoid arthritis (RA) patients.Methods RA-FLS were co-cultured with peripheral blood monocytes (PBMCs) from healthy volunteers in the medium containing M-CSF.Bcl6 protein and mRNA in osteoclasts and their precursors were determined by immunofluorescence and Real-time PCR at day 0,7,14 and 21,respectively.Osteoclasts were identified by tartrate-resistant acid phosphatase (TRAP) staining.Bone resorption activity of osteoclasts was determined by bone slices stained with toluidine blue.Kruskal-Wallis H and Bonferroni were used for statistical analysis.Results ① Immunofluorescence staining and TRAP staining showed that Bcl6 protein was mainly expressed in the nuclei of PBMCs.After co-cultured with RA-FLS for 7 days,some PBMCs differentiated into macrophages and a few differentiated to TRAP-positive multinucleated osteoclasts,and the total Bcl6 protein expression in osteoclasts and their precursors were increased.At day 14,the total Bcl6 protein expression was increased further.At day 21,the Bcl6 protein expression in nuclei of osteoclasts was decreased while PBMCs were differentiated into osteoclasts,and total Bcl6 protein expression was decreased.②Real-time PCR showed that Bcl6 mRNA expression in osteoclasts and their precursors at day 7 tended to increase than that at day 0 (x2=3.429,P＞0.05).At day 14 after co-cultured with RA-FLS,Bcl6 mRNA expression in osteoclasts and their precursors was significantly higher than that at day 0 (x2=5.333,P=0.045).At day 21,the expression of Bcl6 mRNA was significantly lower than that at day 14 (x2=6.023,P=0.038).Conclusion Bcl6 may be involved in osteoclast differentiation and activation,and may play a role in the inflammatory status in the process of differentiation from PBMCs to macrophages.Further studies are needed to establish the mechanisms.

Application of team-based learning in practice teaching of internal medicine / 中华医学教育探索杂志

Objective To explore the feasibility of team-based learning (TBL) for practice teaching of internal medicine.Methods TBL was carried out in 40 interns from clinical medical students of eight-year program.Every team included 4 ～ 5 members.The course of rheumatoid arthritis was chosen according to the course syllabus.Results The students discussed enthusiastically in the team test and in-class application exercises.More than 80％ of the students expressed that TBL was helpful to understand the course content better,to learn the disease better,to elevate personal study ability and to know others' thought processes.76％ of the students supported to carrry out TBL again.Conclusions TBL is helpful for senior medical students to elevate their ability of applying course concepts to solve practical problems through a series of team activities.Improvement of teaching idea and skill is the key to perform TBL successfully.

Liver safety of tumor necrosis factor-α antagonists in inflammatory arthropathy patients with concurrent chronic hepatitis B infection: a prospective observation / 中华风湿病学杂志

Objective To investigate the effect of tumor necrosis factor (TNF)-α antagonists on liver function and reactivation of hepatitis B virus ( HBV ) in patients with inflammatory arthropathy with concurrent chronic HBV infection.Methods Patients with active rheumatoid arthritis (RA) and ankylosing spondylitis (AS) who were grouped according to serum HBV biomarkers were treated with TNF-α antagonist.The liver function and reactivation of HBV were monitored before and after anti-TNF-α therapy.Kruskal-Wallis one-way analysis of variance on ranks of continuous variables and x2 test or Fisher's exact test for categorical variables among 3 or more groups.Results Fifty patients were enrolled with 3 to 23 months of follow-up visit.The level of transaminases in chronic HBV infection group [n=11,AST (36±18) U/L,ALT (44±46) U/L] were significantly higher than that in past HBV exposure group [n=16,AST (22±6) U/L,ALT (17±9) U/L] or free of HBV infection group [n=23,AST (19±6) U/L,ALT (15±9) U/L](AST:x2=11.161,P＜0.01,ALT:x2=8.038,P＜0.01).One patient with elevated baseline HBV-DNA load was treated concomitantly with lamivudine and anti-TNF-α therapy,and the HBV-DNA load reduced about to normal 4 months later.Among the other 10 patients with normal baseline HBV-DNA load in chronic HBV infection group,one patient showed reactivation of HBV with elevated transaminases after anti-TNF-α therapy; another patient had only elevated transaminases without reactivation of HBV,and the transaminases returned to normal after withdrawal of antiTNF-α therapy,which suggested drug-induced liver injury.All patients in both past HBV exposure group and free of HBV infection group remained HBsAg negative after the therapy.Conclusion Patients with inflammatory arthropathy should be screened for HBV infection and check liver function before anti-TNF-α therapy,and carefully monitor the reactivation of HBV and liver function during treatment.Patients with concurrent chronic HBV infection should be treated conco-mitantly with anti-virus and anti-TNF-α therapy if they have elevated baseline HBV-DNA load (＞105 copies/ml,in particular) and good economic situation.

The validity of high-frequency Doppler ultrasound in identifying knees rheumatoid synovitis / 中华风湿病学杂志

ObjectivesTo examine the validity of high-frequency Doppler ultrasound in identifying knees synovitis in patients with rheumatoid arthritis(RA).MethodsNinety-five consecutive patients with active RA were examined withhigh-frequency Doppler ultrasound to examine synovitis signals in knees.Synovial tissue samples of 51 patients were obtained by closed needle biopsy from knees after ultrasound examination.Serial synovial tissue sections were stained with H&E and immunohistochemical staining,and the histopathological synovitis scores were evaluated.The relationship among clinical, histopathological and ultrasound synovitis indexes was analyzed by Spearman's rank order correlation test and receiver operating characteristic curve analysis.ResultsAmong 95 RA patients,the median thickness of synovial membrane in ultrasound was 2.8 mm,the median depth of effusion was 2.7 mm; Doppler signals of synovial blood flow were detected in 82％(78/95 ) of patients and the median semiquantitive grading of synovial blood flow was 1.0.The thickness of synovial membrane and synovial blood flow at Doppler ultrasound correlated positively with histological synovitis score,hyperplasia of the lining layer,and inflammatory infiltration in sublining area (the thickness of synovial membrane:r=0.438,0.424,0.368,respectively; synovial blood flow:r=0.357,0.377,0.347,respectively; all P＜0.05).Although there was no significant difference in clinical synovitis indexes between patients with histologically low-grade and high-grade synovitis,the thickness of synovial membrane and synovial blood flow in ultrasound in patients with histologically high-grade synovitis was significantly higher than those with low-grade synovitis(P=0.001,0.036,respectively).When the thickness of synovial membrane in ultrasound was ≥ 3.9 mm,the specificity of diagnosing the high-grade synovitis was 96.7％ and the sensitivity was 61.9％.ConclusionSynovitis signals at high-frequency Doppler ultrasound correlate with histopathological synovitis,and it might be helpful in evaluating the severity of histopathological synovitis.

Rosiglitazone inhibits receptor activator of NF-κB ligand expression in rheumatoid synovial fibroblasts through p-ERK pathway / 中华风湿病学杂志

ObjectiveTo investigate the effect of rosiglitazone (RSG), a high-affinity synthetic agonist for PPAR-γ, on the expression of receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG)in rheumatoid arthritis fibroblast-like synoviocyte (RA-FLS) and the underlying mechanisms. MethodsRAFLS were incubated with different concentrations of RSG(0, 5, 10, 15, and 20 μmol/L) for 3 days, RAFLS proliferation was detected by Cell Counting Kit-8 assay, and then the expression of RANKL and OPG was examined by real-time PCR and Western blot. The expression of phosphorylated ERK, p38 and JNK was also detected after RA-FLS was incubated with RSG(0, 15, and 20 μmol/L) for 3 days. One-way ANOVA and Bonferroni were used for statistical analysis. ResultsRSG (5, 10, 15, and 20 μmol/L) had significantly inhibited the expression of RANKL mRNA(0.503±0.005, 0.438±0.031, 0.161±0.042, 0.050±0.018) and protein (1.72±0.09, 1.58x±0.05, 1.46±0.11, 1.22x±0.14) in RA-FLS (F=200.820, F=13.602, P＜0.01 ), while the expression of OPG mRNA (2.8 ±0.5, 7.0 ±3.2, 8.4 ±2.3, 25.7 ±5.1 ) and protein ( 1.21 ±0.11, 1.52 ±0.16, 1.63±0.11, 1.91 ±0.03) increased significantly (F=26.531, F=24.872, P＜0.01), both in a dose dependent manner. The ratio of RANKL/OPG mRNA and protein significantly reduced (F=453.425, P＜0.01 ;F=4.173, P＜0.05 ). RSG (15, 20 μ mol/L) significantly inhibited the expression of p-ERK-1/2 protein (0.55±0.06, 0.45±0.06, F=6.991, P＜0.05). ConclusionRSG can inhibit RANKL expression in RA-FLS through p-ERK pathway.