Objective To detect TRAF1 gene single nucleotide polymorphism in Han population ,and analyze its association with increased risk of rheumatoid arthritis (RA) in Han people. Methods The study group was comprised with 100 healthy Han subjects and 100 Han RA patients. Six SNPs was detected by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOFMS), and the result was analyzed by Haploview software. Fisher's exact test and χ2 test were used for statistical analysis.Results The rs4836834 A/A, A/T, T/T allele frequencies of RA group were 22.22%, 49.49%, 28.28% respectively. The rs4836834 A/A, A/T, T/T allele frequencies of healthy control group were 28.57%, 30.61%,40.82% respectively. There was significant difference (P<0.05) of rs4836834 between Hah patients with RA and healthy controls. For other five sites, including rs10818488, which had been reported to be associated with increased risk of Caucasians RA patients, there was no significant difference (P>0.05) between RA group and the control group. The linkage disequilibrium and haplotype frequency analysis results showed that there was strong linkage disequilibrium between rs7021049 and rs7021880, rs7021049 and rs4836834, rs7021880 and rs4836834, but there was no significant difference between RA group and the control group in the haplo-type frequencies of the three classes of linkage disequilibrium. Conclusion TRAF1 gene may be a susceptible gene associated with Han RA people, but its single nucleotide polymorphisms and performance may be different between Caucasians and Han people.

Objective To study whether IL-1β, a catabolic factor of cartilage metabolism, induces premature senescence of articular chondrocytes, and whether caveolin-1 mediates IL-1β-induced cellular senescence. Methods Cultured human articular chondrocytes were stimulated with 10 ng/ml IL-1β. Cellular senescent phenotypes were analyzed by cellular morphology, cell growth arrest (flow cytometry), telomere erosion (Southern blotting), life span (population doublings), and specific senescence-associated β-galac-tosidase (SA-β-Gal) activity. Expression level of caveolin-1 was modulated by anti-sense oligunucleotide or transfection of caveolin-1 gene. Caveolin-1 protein was analyzed by Western blotting. Results Incubation of chondrocytes with IL-1β markedly increase the percentage of cells in G0/G1 phase and reduce the percentage in S phase. Single stimulation with IL-1β enables chondrocytes to become big and flat, and SA-β-Gal activity in chondrocytes is enhanced. Repeated stimulation with IL-Iβ resulted in accelerats erosion of mean telomere length, and shortens life span. Down-regulation of caveolin-1 with anti-sense oligonucleotide significantly inhibits the features of chondrocytes senescence induced by IL-1β. In contrast, caveolin-1 overexpreasion enhanced SA-β-Gal activity in the chondrocytes. Conclusion IL-1β induces features of stress-induced premature senescence and telemere-dependent replicative senescence of articular chondrocytes, which is mediated by caveolin-1. These data suggest that IL-1β induces premature senescence of articular chondro-cytes by upregulation of caveolin-1, which facilitates the development of osteoarthritis.

Objective To explore the metabolic characteristics of rheumatoid arthritis (RA) patients with type 2 diabetes (T2DM) and provide evidence for the management of cardiovascular risk factors. Methods One hundred and four RA patients with T2DM and 100 healthy subjects with matched age and sex were the subjects of study. The metabolic parameters of the two groups was compared and the ratio of metabolic abnormalities in RA with T2DM group was analyzed. Comparisons between groups were analyzed by t-test and Chi-square analysis. Results The average duration of RA and T2DM were (8±6) and (10±5) years respectively; 55.8% patients with CRP>10 mg/L and 72.1% patients with ESR>30 mm/1 h. There was no significant difference in body mass index between the two groups [(23.3 ±3.1) kg/m2 vs (23.4 ±2.8) kg/m2, P=0.991]. The systolic blood and diastolic blood pressures of RA patients with T2DM were significantly higher than those of the control group (P<0.01). There was no significant difference in blood uric acid [(0.27 ± 0.11) mmol/L vs (0.27 ±0.12) mmol/L, P=0.957]. There was no significant difference in the levels of total cholesterol (TC) [(4.6 ±1.0) mmol/L vs (4.5 ±0.5) mmol/L, P=0.547], but the levels of triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) [TG (1.4±0.8) mmol/L vs (1.1 ±0.3) mmol/L, t=2.871, P=0.005; HDL-C (1.1 ±0.3) mmol/L vs (1.5 ±0.4) mmol/L, t=-7.064, P<0.01;LDL-C (2.6±0.8) mmol/L vs (2.4±0.4) mmol/L, t=2.003, P=0.047] were significantly different in the two groups. 36.5% patients were with hypertension, 17.3% patients were with high TC, 30.7% patients were with high TG, 26.9% patients were with low HDL-C, and 27.8% patients were with high LDL-C. Conclusion High incidence of hypertension, poor blood sugar control, and lipid metabolism disorders are prominent metabolic disorders in RA patients with T2DM. Clinicians, particularly rheumatologists, need to give adequate attention to these conditions.

Objective To analyze the clinical features of eosinophilic granulomatosis with polyangiitis (EGPA).Methods The clinical characteristics,laboratory test,therapeutic regimen,pathology and imaging diagnosis in 23 cases with EGPA from the First Affiliated Changhai Hospital of Second Military Medical University within June 2007 to January 2013 were retrospectively analyzed.Results There were 14 males and 9 females,with the age ranged from 21 to 68 years.The mean age was (42±9) years.The overall prevalence of asthma was 78％(18/23).The allergic rhinitis accounted for 61％(14/23).The maxillary sinuses were the most frequendy involved,which accounted for 57％(13/23).Skin involvement was 57％(13/23),peripheral neuritis was 70％(16/23).Central nervous system involvement presented cerebrovascular event.Cardiac involvement accounted for 48％(11/23),digestive system involvement accounted for 17％(4/23).The outcome of auxiliary laboratory test revealed that 96％(22/23) patients expressed significantly higher levels of IgE and 70％(16/23) patients carried anti-neutrophil cytoplasmic antibodies (ANCA) which were presented as the perinuclear ANCA (p-ANCA).The patchy infiltrates of lung CT scan accounted for 69％ (11/23).EMG showed mononeuritis multiplex and symmetric sensory motor neuropathy.The abnormal ratios of ECG and Echocardiography were 48％(11/23),79％(11/14) respectively.The pathological manifestations were necrotizing vasculitis,eosinophilic tissue infiltration,and extravascular granulomas.Conclusion Our results confirm the heterogenicity in clinical presentation and lack of specificity.Early diagnosis and treatment will be helpful for good prognosis.

Objective To construct the stably over-expressing hB7-H3 gene human pancreatic cancer cell line PANC1,and provide tools for further investigating the function of hB7-H3.Methods hB7-H3 gene fragment was inserted into lentiviral plasmid GV287 carrying GFP to construct recombinant hB7-H3-GV287 plasmid vector.293T cells were transfected,and the GFP expression was evaluated under fluorescence microscopy.Western Blot wasused to detect the expression of hB7-H3 protein.Lentiviral vectors were packaged and the titer was determined.The recombinant hB7-H3 expressed lentivirus was used to infect PANC1 cell.Flow cytometry was applied to detecte the percentage of GFP and hB7-H3 positive cells.Real-time PCR and Western Blot was used to verify the mRNA and protein expression of hB7-H3.Self-cyclizing GV287 plasmid served as negative control (NC).Results PCR amplified fragment of recombinant plasmid was around 1 368 bp,and no amplified production of NC plasmid was observed.The DNA sequencing of recombinant plasmid was completely consistent with the designed fragment,indicating that hB7 H3-GV287 plasmid was successfully constructed.293T cells transfected with recombinant plasmid expressed hB7-H3 protein,while those cells transfected with NC plasmid did not express hB7-H3 protein.The virus titer of lentiviral packaged recombinant hB7-H3 plasmid was 2 × 108 TU/ml.The percentage of hB7-H3 positive cells,hB7-H3 mRNA and protein expression in PANC1 cells infected with cells infected with hB7-H3 lentivirus was 94.3％,5.09 ± 0.24 and 2.85 ± 0.27,respectively,which was obviously higher than 18.5％,1.28 ± 0.53 and 0.44 ± 0.69 in cells infected with NC lentivirus,and the differences were statistically significant (P value ＜0.01).Conclusions A human pancreatic cancer cell line PANC1 stably over-expressing hB7-H3 was successfully constructed.

Objective To investigate the role and mechanism of RSPO1 in osteoblasts differentiation.Methods The xCELLigence system was used to study the toxicity and role of RSPO1 on the C2C12 cells proliferation.Alkaline phosphatase (ALP) activity was measured by using a phosphatase detection kit.The expression of osteoprotegerin (OPG) was determined using enzyme-linked immunosorbent assay (ELISA).Wnt/β-catenin signaling was evaluated using the TOPflash T cell factor (TCF) luciferase reporter assay.Western blotting assay was performed to confirm the accumulation of β-catenin protein.T test was used for statistical analysis.Results RSPO1 had no effect on the C2C12 cells proliferation,and it produced no toxicity to C2C12 cells.RSPO1 alone resulted in a slight increase in the activity of ALP (2.85±0.08 vs 1.74±0.21,t=3.014,P＜0.05) and the expression of OPG (1.29±0.13 vs 1.00±0.21,t=3.348,P＜0.05),whereas the combination of RSPO1 and Wnt3A significantly amplified ALP activity (81.37±5.08 vs 1.74±0.21,t=31.31,P＜0.01) and OPG protein expression (5.26±0.60 vs 1.00±0.21,t=6.99,P＜0.01).RSPO1 synergized with Wnt3A to activate TCF activity and induce accumulation of β-catenin (3.28±0.18 vs 1.00±0.21,t=10.94,P＜0.05).However,RSPO1 alone had no effect on the TCF activity and β-catenin accumulation (1.25±0.08 vs 1.00±0.21,t=2.225,P＞0.05).Conclusion Our study has revealed that RSPO1 synergized with Wnt3A in activating Wnt/β-catenin signaling pathway to induce osteoblasts differentiation,which demonstrate the therapeutic potential of RSPO1 for RA.

Objective To investigate the effects of Dishevelled (DVL) on apoptosis of diffuse large B-cell lymphoma (DLBCL) cell line OCI-Ly10, and to explore its possible mechanism. Methods Lentivirus plasmid overexpressing DVL2 was constructed, and after virus was packaged, it was transfected into OCI-Ly10 cells. Flow cytometry was used to detect the apoptosis rate of OCI-Ly10 cells with or without the stimulation by TNF-α recombinant protein. Then the gene expression of anti-apoptotic genes, GADD45β and A20, in NF-κB pathway was detected by RT-PCR. Results The virus was sucessfully transfected into OCL-Ly10 cells which overexpressed DVL2. The apoptosis rate of OCL-Ly10 cells overexpressing DVL2 without the stimulation by TNF-α was increased compared with that of the negative control group [(15.46 ±2.37) % vs. (11.72±3.53)%, P=0.03], the A20 mRNA expression level was decreased compared with that of the negative control group [(0.66 ±0.01) vs. 1, P=0.04], and the relative expression level of GADD45β mRNA was not significantly decreased compared with that of the negative control group [(0.79 ±0.15) vs. 1, P=0.642]. The apoptosis rate of DVL2 overexpression OCI-Ly10 cells stimulated by TNF-α was significantly higher than that of the negative control group treated by TNF-α [(22.78±4.56)%vs. (12.79±2.89)%, P=0.007]. The gene expression of A20 and GADD45β in DVL2 overexpression cells stimulated by TNF-α was significantly increased, however, the magnitude of increase in DVL2 overexpression cells was less than that in the negative control group treated by TNF-α [A20: (3.75 ±0.14) times vs. (6.89 ±0.10) times, P=0.008; GADD45β:(4.750±0.21) times vs. (6.14±0.08) times, P=0.03]. Conclusion DVL can promote the apoptosis of OCI-Ly10 cells, and its mechanism may be related with anti-apoptotic genes that inhibits its downstream via NF-κB pathway.

Objective To analyze quality of life (QOL) and its related factors in community middle-aged and aged people with osteoporosis (OP). Methods A total of 152 OP patients in one community in Shanghai were surveyed and their QOL was evaluated with a questionnaire integrated with the QUALEFFO-41 questionnaire. Factors were analyzed using a Kruskal-Wallis test and stepwise multiple regression. Results The QOL scores of these middle-aged and aged people with OP were generally low (62.58 + 14.06). Age, education level, milk intake, a history of fractures or illness, OP grade and menopause time were significant factors predicting QOL scores. Stepwise multiple regression indicated that fractures, illness, education level and menostasis time had a linear relationship with QOL. Conclusion The key points in improving the QOL of middle-aged and aged people with OP in the community are health education, early diagnosis and comprehensive treatment.

BACKGROUND:Studies have funded that reduced Wnt/β-catenin signaling is involved in the onset and/or progression of bone erosion in rheumatoid arthritis. It can lead to potential new treatment approaches of bone erosion by enhancing Wnt/β-catenin signaling pathway. R-spondin 1 may act as a Wnt agonist, but there is no study in human osteoblasts. OBJECTIVE:To verify the effect of R-spondin 1 on promoting differentiation and maturation of human osteoblasts by inhibiting DKK1. METHODS:S40-transfected human osteoblast lines, hFOB1.19, were treated with R-spondin 1, Wnt-3a and DKK1 to detecting the proliferation, alkaline phoshpatase activity and osteoprotegerin concentration. RESULTS AND CONCLUSION:R-spondin 1 had no effects on hFOB1.19 cel s. Wnt-3a upregulated the activity of alkaline phoshpatase, which could be enhanced by addition of R-spondin 1. R-spondin 1 could reduce the DKK1-mediated inhibition of alkaline phoshpatase activity in hFOB1.19 cel s. R-spondin 1 increased the concentration of osteoprotegerin, and moreover, the promotion of osteoprotegerin by R-spondin 1 alone was stronger than the inhibition by DKK1. These findings suggest that R-spondin 1 can inhibit DKK1 by Wnt/β-catenin signal pathway to promote the differential and maturation of human osteoblasts to excrete osteoprotegerin.

Disease-associated autoantibodies (AAB) are important for the diagnosis of respective autoimmune diseases (AID).Autoantibodies can also be used for monitoring of response to therapy and for prognostic purpose.However,significant biological heterogeneity of autoantibody response,the difficulty in simultaneously improving detection sensitivity and specificity of autoantibodies and the lack of standardization in detection methods lead to limitations in its clinical applications and some difficulties in explaining the test results.It is important to search for novel autoantibodies in sera,to establish and standardize automated detection platforms with good quality and to perform well-designed clinical evaluation in the future research and clinical applications of autoantibodies.