Objective:To evaluate the impact of altered collagen Ⅱ(CⅡ) peptide(S268-270) on T cell activation.Methods:Altered CⅡ263-272 with G268,P269 and K270 consecutively substitution with A or G were synthesized using solid phase various concentrations were added to HLADR1 transfected APC.Expression of FITC stainning was analyzed by fluorescence-activated cell sorting,and the binding of altered CⅡ peptide to HLA-DR1 molecule on cell membrane was evaluated.Irridiated HLA-DR1 expressing APC was incubated with CⅡ263-272 or hemagglutinin(HA)306-318 and altered CⅡ peptides at various concentrations for 2 hours before T cells(3.19) were added.Supernatant was harvested and then added to IL-2 dependent CTLL cell.The cell proliferation was examined by methotetrazolium(MTT) method.Results:The altered CⅡ peptide and wild type of CⅡ263-272 were able to competitively bind to HLA-DR1 molecule expressed on cell membrane of transfected APC.CⅡ or HA induced T cell activation was significantly inhibited by the altered peptide S268-270.Conclusion:Altered CⅡ263-272 with G268,P269 and K270 consecutively substitutions with A or G inhibited CⅡ263-272 and HA306-318 induced T cell activation through competitively binding to HLA-DR1,suggesting a new approach in inhibition of T cell activation in RA.

Objective To investigate the effects of 5-Aza-CdR( methylation transferase inhibitor) on the expression levels of leptin gene in chondrocytes and methylation states of leptin promoter region between osteoarthritis (OA) group and control. Methods The chondrocytes in osteoarthritis group were treated with 5-Aza-CdR with different doses and time-points, and the expression level of leptin was detected by real-time polymerase chain reaction for picking up the optimum dose and time-point. Next, the chondrocytes in 5 osteoarthritis patients and 5 control patients (amputation due to severe trauma) were treated with 5-Aza-CdR. Lastly, leptin mRNA expression levels in the four groups osteoarthritis and control chondrocytes treated with/without 5-Aza-CdR were measured by real-time PCR and the methylation state of promoter region ( - 280- + 79) was detected by epityper quantitative DNA methylation analysis. Results ( 1 ) After treating the chondrocytes in OA groups with 10 μmol/L 5-Aza-CdR for 72 h, the mRNA expression levels of leptin were increased significantly. ( 2 ) The mRNA expression levels of leptin were significantly different among the four groups ( P ＜ 0. 05 ), and the chondrocytes in osteoarthritis groups treated with 5-Aza-CdR showed a marked induction of leptin mRNA expression. (3) Analysis of quantitative methylation data using an unsupervised hierarchical clustering algorithm, showed that methylation patterns of leptin promoter was different between control and osteoarthritis chondrocyte treated with/without 5-Aza-CdR. Conclusion Demethylation of leptin promoter might up-regulate leptin gene expression level and it might contribute to osteoarthritis.

Objective This study is to investigate cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway gene expression pattern of peripheral blood mononuclear cell(PBMC) of primary sjgren syndrome patients.Methods The PBMC sample of 3 patients with sjgren syndrome and 3 healthy volunteers with consistent age were collected.The total RNAs was extracted from the PBMC samples,and reverse transcripted in vitro transcription(IVT),labeled with Cy5/Cy3 and hybridized on the gene chips.After scanning and data extraction with LuxScan 3.0,differentially expressed genes were analyzed with SAM method.The online tool of molecule analysis system(MAS) was used for biological knowledge mining.Results Statistical difference was calculated between the patient and control group in the following three pathways: cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway.Among these,genes of IL-2RA,IL-10 were up-regulated and genes of PF4,GZMA were down-regulated.Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjgren′s syndrome.And the research may provide new target therapy for SS.

Six patients with Neuro-Behcet disease (NBD) were admitted in Beijing Hospital from 2000 to 2014,during the same period 344 NBD cases were reported in the literature.The clinical data of 350 Chinese NBD patients,including the symptoms,signs,laboratory tests,imaging,treatment and prognosis were retrospectively analyzed.The average onset age was 36.7 years old.The average interval time from Behcet disease (BD) to NBD was 5.6 years.NBD often occurred in male,the male/female ratio was 1.94:1.The most common manifestations were oral ulcer 94.9％ (277/292),followed by genital ulcer 69.1％ (215/311) and fever 42.1％ (131/311).The most common symptoms in neurology were limb weakness (36.9％,129/350),headache (34.0％,119/350) and sensory disturbance (21.1％,74/350),followed by bulbar symptoms (11.4％,40/350),pyramidal sign was most often (27.1％,95/350).The central neurology system (CNS) was the major target of NBD,parenchymal CNS was affected most (88.0％,308/350),non-parenchymal CNS was affected less (5.7％,20/350),peripheral neurology system (PNS) was rarely affected (6.9％,24/350).Steroids were the first-line drug for NBD,if it failed,immunosuppressive agents or biologic agents could be used.The most NBD patients had single episodes,32.0％ (49/153) had relapses with remission,7.7％ (27/350) had adverse prognosis.Analysis showed that NBD should be paid more attention.

Objective To investigate signal transduction and cytokine gene expression pattern in peripheral blood mononuclear cell (PBMC) between primary Sjogren syndrome patients and healthy controls. Methods The PBMC of 3 patients with Sjogren syndrome and 3 healthy volunteers with consistent age were collected. The total RNA extracted from the PBMC was carried the reverse transcription and in vitro transcription (IVT), then labeled with Cy5/Cy3 and hybridized on the gene chips. After scanning and data extraction with LuxScan 3.0, differentially expressed genes were analyzed with SAM method. The online tools of molecule analysis system (MAS) was used for biological knowledge mining. The difference of signal transduction and cytokine gene expression of each patient and control were compared. Results Statistical difference was calculated between the patient and control group in the following four pathway: the chemokine-cytokine receptor interaction pathway[(FASLG(Fas ligand), CCL5, CCR3, IL-4R, CXCL10 (CXC chemokine ligand 10), TNFRSF19L (tumor necrosis factor supeffamily 19 ligand), CX3CR1 (CX3C chemokine receptor 1)], adhension molecule pathway [interleukin adhesion molecules-1 (ICAM-1)], Jak-STAT signal pathway [STAT4 (signal transducer and activation of transcription 4), CISH (chromogenic in situ hybridization), IL-4R], Toll-like receptor signal pathway(CCL5, CXCL10). Among these, 4 genes were up-regulated and 6 genes were down-regulated. Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjogren syndrome.

ObjectiveTo explore the prevalence and clinical features of hyperuricemia and its influencing factors in elderly male people aged 90 years and above. MethodsOne hundred elderly male people aged 90 years and above who underwent routine health examination in our hospital in 2007 were selected in the study. Serum uric acid level was examined by uricase-peroxidase method, and all patients were divided into hyperuricemia group and control group according to the serum uric acid level. Clinical and biochemical indications were compared between the two groups, and logistic regression was used to analyze the influencing factors of hyperuricemia in elderly people. ResultsThe serum uric acid level was increased in 20% of the elderly people, and the prevalence of gouty arthritis was 1%. The levels of blood urea nitrogen and serum creatinine were higher in hyperuricemia group than in control group[(10. 98±4.29) mmol/L vs. (6. 87± 1.86) mmol/L, (125.2±25.9)μmol/L vs. (93. 4±19. 8)μmol/L;both P<0.05)3. The patients in hyperuricemia group had a higher prevalence of hypertension and hypertriglyceridemia, and a higher proportion of diuretic application than patients in control group(P<0. 05). Logistic regression analysis showed that serum uric acid level had the most remarkable correlation with serum creatinine(OR= 1. 969), followed by fasting blood glucose (OR= 1. 310)and blood urea nitrogen(OR = 1.161). There was negative correlation between serum uric acid level and plasma cholesterol level(OR = 0. 802). ConclusionsThe prevalence of hyperuricemia is high in elderly people aged 90 years and above, while the incidence of gouty arthritis is low. Renal function impairment, metabolic syndrome and thiazide diuretic are the major factors for hyperuricemia.

Objective To identify gene expression profile in periphend blood mononuclear cell (PBMC)of patients with primary Sjogren's syndrome(pSS)complicated with hematology involvement and compare it to healthy volunteers.Methods Three Sjogren's syndrome(SS)patients with leucopenia and 3healthy volunteers were included.The cRNA probes prepared for total RNA were hybridized with three identical gene chips.The difference of gene expression of each patient and volunteers were compared.Results Significant difference in the expression of 82 genes could be detected between patients and volunteers.Among these,45 were upregulated,and 37 were downregulated.Statistical difierence was calculated between patients and volunteers especially in the following two pathways:the complement and coagulation pathways(including C2,PROS1,F2R and SEPPING1)and the cytokine-cytokine receptor interaction pathway(including FASLG,MPL,CCL20 and CXCL2)(P<0.01).Conclusion This study has identified distinct gene expression profiles in PBMC from patients with pSS patients with hematology involvement and healthy volunteers.This provides new knowledge about groups of genes that are upregulated during disease.It may form an excellent platform for future functional studies.

Objective To identify novel markers such as THRA or TCL1 in Sj(o)gren's syndrome (SS) patients to discriminate them from hcalthy volunteers.Methods Experimental group (n=40) and healthy volunteer group (n=40) were recruited based on strict inclusion and exclusion criteria.Peripheral blood samples (9 ml) were collected and peripheral blood mononuclear cells were separated by Ficoll gradient centrifugation.RNA was extracted using Trizol reagent,and the expression of THRA,TCL1 in PBMCs was detected by real-time PCR.And the data were processed with Wilcoxon test and t test (P＜0.05).Results TCL1 and THRA expression level are higher in SS patients (2.5±4.7) than healthy volunteers (Z=-2.045,P＜0.05).The TCLI expression level (4.4±3.8) is higher in high lymphadenopathy activation index patients (2 to 3grade) than that in low lymphadenopathy activation index patients (grade 0 to 1 ) (t=-2.319,P＜0.05 ).Conclusion TCL1 expression level is higher in SS patients,and TCL1 expression level is related to the severity of lymphadenopathy,which provide a new platform of the study for the pathogenesis,disease severity evaluation,even dia-gnosis and treatment of SS.

Objective To investigate the correlation of disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5 gene single nucleotide polymorphism (SNP) and osteoarthritis (OA) in Beijing.Methods In this case-control study 166 OA patients and 204 normal controls were collected from Beijing.Sorted gene type by SNaPshot technique,and analyzed the difference of allele and genotype frequencies between OA and the controls.The correlation of linkage disequilibrium and haplotype of SNPs with OA was analyzed through Haploview softeware.Results Rs2132824 was the positive site with Bonferroni method after multiple testing correction (x2=0.1 1,P＜0.05),of which allele frequencies had no significant difference between OA and the control (P＞0.05).But in codominant genetic model,rs2132824 CT genotype with OA was inversely proportional to the risk of OA [OR (95％CI) was 0.52 (0.32,0.86),P＜0.05],and TT genotype was directly proportional to the risk of OA [OR(95％CI) was 3.16 (1.55,6.47),P＜0.05],while in recessive genetic model,TT genotype was in direct proportion to the risk of OA [OR (95％CI) was 3.99 (1.99,8.01),P＜0.05].Linkage disequilibrium in SNPs of rs151065,rs229077,rs56153501,rs2830580 and rs58215296 did exist,which made haplotype of rs 151065-rs229077-rs56153501-rs2830580-rs58215296 (G-T-A-C-A) directly proportional to the risk of OA [OR(95％CI) was 1.874(1.019,3.446),P＜0.05].Conclusion ADAMTS-5 gene SNP is associated with OA susceptibility risk in Beijing.Genotype CT of rs2132824 is a low risk factor for OA while TT is a hygh risk factor.Haplotype of G-T-A-C-A is connected with high risk of OA.

Objective To detect serum level of thrombospondin-1 (TSP-1) in rheumatoid arthritis (RA) patients,and analyze the correlation between serum TSP-1 level and disease activity in elderly versus young and middle aged RA patients in order to provide the basis for diagnosis and evaluation of RA.Methods Peripheral blood was collected from 98 RA patients (including 33 elderly,65 young and middle-aged RA patients) and 58 healthy controls.Serum TSP-1 levels were detected by enzyme linked immunosorbent assay (ELISA).Difference in TSP-1 level between RA patients and healthy controls was analyzed.Correlations of TSP-1 level with disease activity parameters of number of tender joints and swelling joints,disease activity score (DAS28 score) and erythrocyte sedimentation rate (ESR),levels of C-reactive protein (CRP),rheumatoid factor (RF),immunoglobin G (IgG),immunoglobin A (IgA) and immunoglobin M (IgM) were analyzed in elderly versus young and middle-aged RA patients.Spearman's and Pearson's correlation analysis were performed.Results TSP-1 level was significantly higher in RA patients than in healthy controls (P＜0.01).TSP-1 level was correlated with number of swelling joints and DAS28 score in RA patients (r=0.246 and 0.241,both P＜0.05).In elderly RA patients,TSP-1 level was correlated with number of swelling joints (r=0.377,P＜0.05),meanwhile significantly positive correlations of TSP-1 level with DAS28 score and rheumatoid factor level were observed in young and middle-aged RA patients (r =0.243 and 0.326,both P＜ 0.05).Conclusions TSP-1 may play roles in the development of RA and its determination may benefit evaluating disease activity.In elderly RA patients,TSP-1 level may reflect severity of rheumatoid arthritis and may be a novel biomarker to the evaluation of disease severity.