Objective Infliximab is a kind of recombinant human mouse chimeric anti-tumor necrosis factor monoclonal antibody. Here we aimed to examine the impact of infliximab therapy on RANK/RANKL/OPG system in the peripheral blood of rheumatoid arthritis (RA) patients. Methods Fifty patients with RA were rigorously screened and randomly divided into 2 groups. One group was treated with infliximab (3 mg/kg)and methotrexate (MTX). As control, the other group was treated with MTX alone. Infliximab was administered at weeks 0, 2, 6 and 14. The expression of RANK, RANKL mRNA in the peripheral blood, serum OPG and clinical indicators changes at week 0 and 18 were compared.x2-test or t-test were used for statistical analysis.Results After treated with infliximab, bone damage of joints were slowed down when examined by radiography in RA patients compared with the control group. And the ratio of OPG/RANKL was also decreased in RA peripheral blood (w0: 80.25;w 18: 63.2); (control group w0: 83.37; w18: 30.87)(P＞0.05). Although after the treatment with either MTX alone [w0: (238±15) pg/ml; w18: (118±10) pg/ml] or infliximab combined with MTX [(w0: (223.1±6.2) pg/ml; w18:(162.4±5.5) pg/ml], the serum levels of OPG were all decreased (P＞0.05), the level of OPG in infliximab treatment group was declined slower than those in the control group. Conclusion RA bone destruction can be inhibited by the combination therapy of infliximab and MTX. The mechanism may be partly through the RANK/RANKL/OPG system.

Objective To investigate the expression of the genes correlated with interferon induced genes virus (MX1, OAS1, IFI44) in the peripheral blood leukocytes of patients with systemic lupus erythematosus (SLE), and to evaluate the relationships between the expression levels of these genes and diseaseactivity. Methods The clinical data of 100 SLE patients, 40 non-SLE patients with rheumatic diseases, and 40 normal controls were collected. Peripheral blood samples were collected. Total RNA was extracted and transcribed into cDNA. SYBR green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as △CT value) of MX1, OAS1 and IFI44 in patients with SLE and those in the controls. Comparisons between groups were performed with ANOVA and Spearman correlations. Results ①The △CT value of MX1, OAS1 and IFI44 expression level of the SLE patients (3.4±1.8, 4.2±1.5, 8.8±2.2)was significantly higher than those of the non-SLE patients (2.4±0.4, 3.4±0.7, 5.4±2.1 ) and normal controls (2.3±1.1, 2.6±0.7, 5.2±2.0). ② The △CT value of OAS1 and IFI44 expression level of the SLE patients in severe disease was significantly higher than those of the SLE patients in mild disease and the SLE patients with stable disease. ③The ACT value of OASI and IFI44 were correlated with the SLEDAI scores (r=0.038,0.380). ④ The △CT values of MX1, OAS1 and IFI44 expression level of the SLE patients with arthritis were significantly higher than those of SLE patients without arthritis. ⑤ The △CT value of IFI44 expression level of the SLE patients with lupus nephritis (3.2±2.1,2.2±1.1) was significantly higher than that of the SLE patients without lupus nephritis. ⑥ There was correlations among these genes in SLE patients (P＜0.05). Conclusion The value of MX1, OAS1 and FFF44 expression level of SLE patients is up-regulated. The real time expression levels of OAS1, IFI44 genes are associated with SLE disease activity and there are close correlation among these genes with interferon induce virus-relationed genes (MX1, OAS1, IFI44) in SLE patients.

Objective To investigate the association between interleukin(IL)-1F7 gene (rs3811047)single nucleotide polymorphism (SNP) and ankylosing spondylitis (AS). Methods SNP of IL-1F7 gene (rs3811047) was analyzed in 158 patients with AS and 181 healthy controls by ligase detection reaction based on high temperature ligase (LDR-PCR). The distribution of IL-1 F7 (rs3811047 ) genotypes and allele frequencies were detected between the two groups. Results Significant differences were found in the distribution of IL-1F7(rs3811047 ) genotypes and allele frequencies between AS patients and healthy controls. The frequency of A allele of IL-1F7 gene at positions rs3811047 was 12.03% and 17.68% in AS group and the control group,and the frequency of G allele was 87.97%, 82.32%, respectively (x2=4.2204, P=0.0399). The percentage of AA, AG and GG genotype was 0, 24.05%, 75.95% in AS group, which differed from the controls group (2.76%, 29.83%, 67.41% ). The difference had remarkable statistical significance (x2=6.2675, P=0.043).The positive rate of HLA-B27 in AS patients which represented as AG genotype was 70.27% (26/37),remarkably lower than that in AS patients which represented as GG genotype 94.23% (98/104), the difference was statistically significant (x2=2.168, P=0.030), erythrocyte sedimentation rate and C reactive protein levels were conspicuously lower than that in GG genotype (t=2.971, P=0.013; t=3.300, P=0.001 ). Conclusion Our study suggests that the SNP (rs3811047) of IL-1F7 may be a susceptibile factor for AS in Anhui Han population, the genotype may influence the clinical phenotypes of AS.Patients who carry the A allele may have less inflammation than patients who do not carry the A allele.

Objective To study the (-)-epigallocatechin-3-gallate (EGCG) function on the proliferation of T cells derived from the peripheral blood and synovial fluid of rheumatoid arthritis (RA) and RA-related cytokine levels and the role of EGCG on RA synovial fibroblasts (FLS) proliferation was investigated. Methods ① Mononuclear cells from RA peripheral blood (30 cases) and synovial fluid (23 cases)were isolated. Blank group, negative control group, positive control methotrexate (MTX) group and therapeutic group with three different concentrations of EGCG were set up. Incorporated isotope 3H was used to test T cell proliferation from RA-PBMC and SFMC. ELISA assay was used to test cytokine (TNF-α, IFN-γ, IL-1, IL-6 and IL-17A) levels. ② MTT assay was used to test FLS proliferation from RA synovial tissue (8 cases).Results ① The CPM value of the high-dose group of EGCG in the peripheral blood and synovial fluid of RA patients was [ ( 15 136±2910), ( 11 587±3135 ) ], which was declined significantly than the control group (42856±2127) (P＜0.01). The levels of TNF-α, IFN-γ, IL-1, IL-6 and IL-17A in the high-dose group of EGCG in the peripheral blood were [(321±13), (298±20), (132±12), (197±7), (59±8) pg/ml], which were decreased significantly than those of the control group [ (458±28), (505±26), (346±28), (405±25),(109±13) pg/ml ] (P＜0.05 or P＜0.01 ). The levels of TNF-o, IFN-γ, IL-1, IL-6 and IL-17A in the highdose group of EGCG in the synovial fluid were [(41.4±2.9), (182±16), (56.3±11.0), (34.2±1.9), (44±8)pg/ml ], which was decre-ased significantly than the control group [ ( 388.3± 19.3 ), (469±20), ( 104.2±17.8 ),( 114.5±4.8), ( 104±11 ) pg/ml] (P＜0.05 or P＜0.01 ). ② The level (A) of the high-dose group of EGCG in the FLS was (0.08±0.02), which was declined significantly than the blank group (0.27±0.04) (P＜0.05).Conclusion ① In vitro EGCG can inhibit T cell proliferation from peripheral blood and synovial fluid of RA patients and the TNF-α, IFN-γ, IL-1, IL-6 and IL-17A secretion are decreased. ② In vitro EGCG can inhibit the proliferation of RA FLS.

Objective To identify the susceptibile genes in a rat model for rheumatoid arthritis (RA),to determine whether sex affects disease onset and to define the mechanisms that impacts congenic genes on arthritis. Methods Arthritis-susceptible DA rats were compared with sex/age-matched congenic rats in which alleles were substituted with alleles from arthritis resistant PVG rats. Incomplete Freund's adjuvant (IFA) was injected from the base of the tail. Arthritis was visually scored, the messenger RNA (mRNA) levels of congenic genes and cytokine were determined by reverse transcription-polymerase chain reaction. The differences between two groups were analyzed using Mann-Whitney U test. Results In oil-induced arthritis (OIA), male congenic R16 rats deviated profoundly from DA rats by decreased arthritis severity (5.9±3.8 vs 9.3±2.3, P＜0.05 ), and markedly reduced lymph node mRNA levels for calsyntenin-3 (Clstn3) gene (0.7±0.4 vs 2.2±1.6, P＜0.01 ) and interleukin (IL)-17 (1.4±2.2 vs 2.7±2.9, P＜0.05) and IL-1β (1.5±2.1 vs 2.3±2.5,P＜0.05) levels. Conclusion Rat Clstn3 gene regulates the production of pro-inflammatory cytokines of OIA in male rats. The effect of arthritis-susceptible gene Clstn3 is gender-specific.

ObjectiveTo investigate the clinical and laboratory characteristics of systemic sclerosis (SSc) patients with digital ulcer(DU) in China.MethodThe data of 166 consecutive SSc patients in EUSTAR DATABASE in Peking Union Medical Colloge Hospital from February 2009 to August 2010 were prospectively collected,and patients with DU were compared with those without DU.All patients fulfilled the ACR classification criteria for SSc in 1980.Results① Forty-nine patients (29.5％) had DU in 166 SSc patients.The disease onset age was(36±12) years(8.1-61.7 years) for those patients with DU.All had Raynaud's phenomenon(RP).② Demographic data:there were significant differences between patients with and without DU in sex (F/M 40/9 vs 112/5,P=0.005),age [(40±12) years old vs(46±12) years old,P=0.005],the onset age of RP [(33±12) years vs(39±13) years,P=0.005] and the duration from RP to the first non-RP presentaion[ (18±15) months vs(115±307) months,P=0.002 ].③ Clinical manifestations and laboratory findings:there were more diffuse SSc patients and more esophageal involvement in patients with DU (P＜0.05).ConclusionsDU in SSc patients is common,especially in man and patients with diffuse SSc.SSc patients with DU usually are younger when RP onsets and the non-RP manifestations usually present earlier when compared with those patients without DU.

ObjectiveTo investigate the association of BANK1 single nucleotide polymorphisms (SNPs) with rheumatoid arthritis(RA) in Chinese Han. MethodsTwo hundreds and twenty-one RA patients and 310 healthy controls who were Chinses Han population from Huashan Hopital and Changzheng Hospital in Shanghai,China were included.DNAs were extracted from peripheral whole blood for study.Samples were genotyped for three variants rs10516487,rs17266594 and rs3733197 in BANK1 by unlabelled probe high resolution melting (HRM) assay.The genotype frequencies of the detected polymorphisms were analyzed in relation to RA and the production of autoantibodies in RA patients.ResultsThe Tr genotype frequency was much higher in RA patients than in healthy controls(X2=6.241,P=0.044).The frequencies of rs10516487 G allele,rs17266594 T allele and rs3733197 G allele were increased among RA patients compared with healthy controls,although they didn't reach statistical significance.The rs10516487 and rs17266594 were found in strong linkage disequilibrium(D'=0.993,r2=0.985).And also the major TGG haplotype of 3-SNP was significantly associated with RA patients[P=0.037,OR =1.345,95％CI (1.018-1.776)].ConclusionBANK1 rs17266594 polymorphism is susceptible to RA,while rs10516487 and rs17266594 are linked in Chinese Han population.BANK1 SNPs TGG haplotype may contribute to RA susceptibility,too.

Objective To investigate the long-term efficacy of nonmyeloablative autologous peripheral blood stem cell transplantation(NAST) to cure refractory autoimmune disease(AD).MethodLong-term follow up of four cases of AD patients with NAST were summarized.The pretreatment regimen was intravenous injection of cytarabin (200 mg· kg-1· d-1 ) and cyclophosphamide (40 mg· kg-1· d-1).The therapeutic effect was evaluated by the change of symptoms and signs and long term complications.Changes of immune function were detected by flow-cytometry.ResultsFive cases of patients had been successfully engrafted.The average time for peripheral leucocytes count to reach 4.0×109/L was 12 days.It needed 10 days for platelets to return to 100×109/L and 22 days for hemoglobin to 120 g/L.Apparent remission of symptoms and signs was observed after transplantation.Lymphocyte subtypes analysis pre- and post- NAST showed that count of CD4+ and the ratio of CD4 +/CD8 + was returned to normal.One patient gave birth to a healthy baby four years after transplantation.Three female patients returned tonormal life. Conclusions Compared with classical myeloablative stem cell transplantation,NAST has a rapid hematopoietic recovery and good long-term therapeutic effect in AD.The quality of life in AD patients treated with NAST is higher than those treated with myeloablative hematopoietic stem cell transplantation.

ObjectiveTo investigate the effects and mechanisms of IL-32γ-shRNA-mediated gene silencing on the proliferation and apoptosis of fibroblast-like synoviocytes in patients with rheumatoid arthritis (RA).MethodsA eukaryotic expression plasmid of shRNA targeting IL-32γ was transfected into fibroblastlike synoviocytes by liposome in patients with rheumatoid arthritis.RT-PCR was used to determine the expression level of IL-32γ.Western blotting was used to detect the levels of cyclin D1 and p-Akt.The proliferation of RA-FLS was examined by MTT.Cell cycles were analyzed by flow-cytometry.The apoptosis of cells were measured by TUNNEL.Comparisons between groups were tested by t test.Results ① The expression of IL-32γwas significantly inhibited by shRNA-IL-32γ-expressing plamid PGCsi 3.0 targeting sequence 1,2 and 3,and the inhibition rate had reached 75.6％,66.2％ and 64.1％,respectively.② The absorbance value of proliferation of RA-FLS in EASY-shRNA-IL-32γ group was significantly lower than that in the shRNA-control group and normal group at day 3 [(0.23±0.03) vs (0.35±0.03) and (0.36±0.04),P＜0.05] and 5 [(0.27±0.03) vs (0.52±0.05) and (0.53±0.04),P＜0.01 ] after transfection.③ The rate of RA-FLS at phase G1 in the EASY-shRNA-IL-32γ group was significantly higher than that in the shRNA-control group and normal group respectively [(88±6)％ vs (69±5)％ and (68±4)％,P＜0.05],while those at phase S+G2 in the EASY-shRNA-IL-32γgroup was significantly lower than that in the shRNA-control group and normal group [ ( 13.6±3.0)％ vs (30.2±4.1)％ and(32.1±4.3)％,P＜0.01].④The rate of RA-FLS apoptosis in the EASY-shRNA-IL-32γ group was significantly higher than that in the shRNA-control group and normal group[(20.50±3.21 )％ vs (9.20±0.32)％ and (8.60±0.22)％,P＜0.01].⑤ The expression of cyclin D1(0.36±0.04) and p-Akt(0.31±0.03) in the EASY-shRNA-IL-32γ group was significantly lower than that in the shRNA-control group [ (0.59±0.08) and (0.53±0.06)] and normal group [(0.61±0.07) and (0.52±0.06),P＜0.01].ConclusionEASYshRNA-IL-32γ can inhibit RA-FLS proliferation by down-regulating the expression of cyclin D1 and induce RA-FLS apoptosis by down-regulating the expression of p-Akt.

ObjectiveTo survey the incidence of depression in rheumatoid arthritis (RA) and explore the related factors.MethodsOne hundred and fifty-nine patients with RA were investigated.All of them were assessed by depression scale and Hamilton Depression Rating Scale.A mono-variate and multivariate Logistic regression analysis were carried out to determine the factors that best related to the occurrence of depression in RA.ResultsThe overall incidence rate was 39.6％.The regression analysis showed that factors related to the occurrence of depression in RA were HAQ-DI (OR=3.276,95％CI 1.315-7.814,P=0.003),the number of tender joints (OR=2.252,95％CI 1.117-3.362,P=0.029),low-income families (OR=1.629,95％CI 1.215-2.437,P=0.031 ) and serum CRP level (OR=1.528,95％CI 1.112-2.294,P=0.040).ConclusionDepression is common in patients with RA.Patients who havehigh HAQ-DI,CRP and from low-income families with more tender joints tend to develop depression.