Objective:To investigate the expression level of plasma miR-320c in patients with osteoarthritis(OA), and explore the clinical significance and the role in pathogenesis of OA.Methods:The clinical data and peripheral blood of 30 patients with OA, 30 patients with connective tissue diseases and 30 healthy control individuals were collected.The levels of plasma miR-320c were detected byfluorescentquantitative reverse transcription PCR(qRT-PCR). Correlation analysis was used to explore the correlation of plasma miR-320c level with knee X-ray data and VAS pain score in OA patients.Finally the miR-320c mimic, the miR-320c inhibitor, and the control material were transfected to the chondrocyte HC-a.The proliferative capacity of HC-a chondrocytes was examined at different time points as determined by the CCK-8 assay.Results:The expression level of plasma miR-320c was significantly higher in OA group(3.26±0.55)than that in the connective tissue diseases group(1.62±0.50)and in healthy control group(1.21±0.66)( F=107.66, P<0.001). Plasma miR-320c expression was positively correlated with radiographic grade( r=0.830, P<0.001), and had no correlation with VAS pain score in OA group( P>0.05). Through repeated measurement variance analysis, the time effect, the group effect and the interaction effect between group and time showed statistically significant differences in chondrocyte proliferation between NC mimic group and miR-320c mimic group( Ftime=5256.767, Fgroup=1947.436, Ftime×group=114.314, all P<0.001). The level of proliferation was significantly reduced.Apoptosis rate of chondrocytes was significantly increased in the group transfected with miR-320c( t=7.85, P<0.01). Conclusions:The expression level of plasma miR-320c is significantly higher in osteoarthritis patients and associated with knee radiographic severity grade.Furthermore, over-expression of miR-320c could suppress the proliferation of chondrocytes.Plasma miR-320c might be potential bio-marker for osteoarthritis knee severity assessment, and involves in regulating chondrocyte growth in the pathogenesis of osteoarthritis.

Anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitis (AAV) is a group of systemic small vasculitis characterized by ANCA positive in serum. Three diseases are included in this group of diseases: granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA) and eosinophilic granulomatosis with polyangiitis (EGPA). In China, standardized diagnosis and treatment of AAV is still lacking. Based on the evidence and guidelines from China and abroad, the Chinese Rheumatology Association formulated the standardization of diagnosis and treatment of ANCA associated vasculitis. The purpose is to standardize the diagnosis of AAV and disease activity assessment, and recommend the treatment strategies.

Objective:To explore the characteristics of plasma microRNA (miRNA) profiles and bioinformatics in patients with osteoarthritis(OA) in order to search for diseases related biomarkers.Methods:Blood samples from 20 cases of OA patients and 15 cases of normal control (NC) were collected to extracted total RNA in plasma. The plasma miRNA expression profile was tested by using Agilent Human miRNA array. Target gene analysis and clustering analysis were performed on differentially expressed microRNAs. Three differentially expressed miRNAs (miR-134-5p, miR-320c and miR-940) were detected by real-time quantitative polymerase chain reaction (RT-qPCR) for further confirmation of microarray data. The differences were tested using t test analysis. Results:① MiRNA microarray showed that compared with NC, there were 74 differential expression genes in plasma of patients in the OA group (FC≥2, P≤0.01), among which 45 were up-regulated and 29 were down-regulated. ② A total of 2 731 potential target genes were predicted in three database, and involved in 462 Kyoto Encyclopedia of Genes and Genomes (KEEG) pathways. Target gene ontology (GO) functional clustering found that the main functions of miRNAs were intercellular adhesion, collagen synthesis, intracellular signal transduction, etc. The main KEGG pathways of miRNAs include mitogen-activated protein kinase (MAPK) signaling pathway, cyclic adenosine monophosphate (cAMP) signaling pathway, osteoclast differentiation signaling pathway, etc. ③ The expression level of miR-20a-5p and miR-320c in OA group were significantly higher than that in controls ( t=-6.142, P<0.05; t=-3.854, P<0.05), while miR-940 was significantly lower than that of controls ( t=2.767, P<0.05) . The trend was consistent with the microarray data. The receiver operating characteristic curve (ROC) curve analyses showed that they were useful biomarkers for differentiating patients with OA from controls. Conclusion:The study shows that plasma in OA patients has a specific miRNAs expression, and miRNAs play an important role in the pathogenesis of OA.

Objective:To study the clinical characteristics and compliance of early-onset gout patients by case-control analysis.Methods:A total of 111 early-onset patients (onset age ≤35 years old) were included as Group A, and 111 non-early-onset patients (onset age >35 years old) with matched disease durationwere included as Group B. The differences ofclinical characteristics, causes of acute gout attack, dairy diet habits, compliance, and misunderstanding of the disease were compared.Results:Compared with the non-early-onsetgoutpatients, the early-onset patients had a higher proportion of obesity (63 cases vs 28 cases), family history (36 cases vs 20 cases) and tophus (39 cases vs 23 cases) and higher level of VAS scores (8.5±1.3 vs 7.6±1.7; χ2=22.988, P<0.01; χ2=5.749, P=0.016; χ2=5.729, P=0.017; t=4.639, P<0.01), lowerproportionof the first metatarsophalangeal joint involvement as the initial joint involvement (45.9%, 51 cases vs 59.4%, 66 cases; χ2=4.066, P=0.044), higher proportion of the ankle involvement as the initial joint involvement (34.2%, 38 cases vs 21.6%, 24 cases; χ2=4.386, P=0.036), higher proportion of alcohol drinkers and high fructose drinkers, which was more likely to relate to alcohol intake, strenuous exercise and high fructose intakeas trigger of the flare ( χ2=6.513, P=0.011; χ2=7.126, P=0.008; χ2=1.978, P=0.160), while the proportion of regular exercisers and on diet in the family was lower ( χ2=22.887, P<0.01; t=-4.917, P<0.01). The proportion of poor diet and medication compliance in Group A was higher than that in Group B(57.7%, 64 cases vs 38.7%, 43 cases; χ2=5.207, P=0.022; χ2=5.867, P=0.015). As for the reason for poor treatment compliance, early-onset gout patients were more worry about the side-effects of drugs than non-early onset patients ( χ2=4.190, P=0.041). There was no significant difference between the two groups in the main misunderstanding of gout. Conclusion:Although early onset gout patients are young, their condition is more serious, and compliance is poorer, this group of patients should be highly valued in clinical diagnosis and treatment.

Objective To recapitulate the clinical traits of conjunctiva-corneal carcinoma in situ with microinvasion,and explore the early diagnosis and precise therapeutic methods.Methods The clinical features,diagnosis,surgical methods and follow-up results of a hospitalized patient with keratoconjunctival in situ carcinoma with microinvasion were retrospectively analyzed and related literature was consulted.Results Surgical resection of the tumor and 2 mm marginal tissue outside the edge of the tumor was performed.Fresh corneal lamellar grafts were used to treat the disease.After 2 years of follow-up,the operation was effective and no recurrence was found.Conclusion Early diagnosis plays an important role in improving the prognosis of keratoconjunctival carcinoma in situ with microinvasion and preventing recurrence.For patients with keratoconjunctival carcinoma in situ acompanied with microinvasion,we have to expand the surgical resection of the tumor (≥2 mm at the edge of the tumor),and follow-up at regular intervals,if necessary,supplemented with chemotherapy or anti-metabolic treatment.

Objective To explore the effect of different concentrations of iguratimod (IGU) on mouse model of bleomycin-induced pulmonary fibrosis.Methods A total of 108 female C57BL/6 mice were randomly divided into the control group,the model group,the low dose IGU group,the moderate dose IGU group,high dose with group and the methylprednisolone (MP) group (n=18 in each group).The mice in the control group were injected with 0.2 ml normal saline endotracheally,and others were injected with 0.2 ml bleomycin (5 mg/kg) from endotracheally to induce pulmonary fibrosis model.The next day,the mice in both control group and the model group were fed with 0.2 ml normal saline every day;The mice in the IGU groups and methylprednisolone group were fed with 0.2 ml iguratimod liquid the IGU (10,30,90 mg/kg) and 0.2 ml methylprednisolone (10 mg/kg) every dayrespectively.Finally the mice were sacrificed at day 7,day 14,day 28 respectively,and the lung tissue was examined by HE staining and Masson staining to evaluate the degree of alveolitis and fibrosis.Repeated measurement of variance analysis was used to compare the differences for time and group,and multi-factor analysis of variance LSD test was used for the comparison between groups.Results ① The body mass of mice in bleomycin-induced groups were decreased compared to the control group.② The pathological alveolitis scores in the high dose IGU group and methylprednisolone group were significantly decreased compared to those of the model IGU group at day 7 and day 14 (P＜0.05),and the pathological fibrosis scores were decreased dramatically compared with the model group at day 14 and day 28 (P＜0.05).Conclusion High concentration of IGU and methylprednisolone can reduce and inhibit lung inflammation and fibrosis of bleomycin-induced pulmonary fibrosis in mice.

Objective To evaluate the clinical performance of chemiluminescent immunoassay (CLIA) on anti-nuclear antibody(ANA) specific autoantibodies testing.Methods A multi-center clinical study A total of 811 Sera samples were collected from 6 collaborating hospitals during the period of April to July 2016, and tested with CLIA and line immunoassay (LIA) in parallel for autoantibodies to ribonucleoprotein(RNP), smith antigen(Sm), SSA/Ro60,SSB/La, centromere protein B(CENPB), double-stranded DNA(dsDNA), nucleosome(Nuc), and ribosome P protein(Rib-P).The positive rate,specificity and qualitative coincidence rate for each antibody between CLIA and LIA methods were analyzed.All discrepant samples for systemic lupus erythematosus (SLE) highly specific autoantibodies (including anti-Sm, dsDNA, Nuc and Rib-P) were retested by enzyme linked immunosorbent assay (ELISA) and further analyzed with SLE disease cohort using McNemar test.Results The positive rate and specificity of CLIA and LIA for antibodies to ANA specific antigens were comparable.Excellent qualitative coincidence were found between CLIA and LIA for the detection of anti-RNP, SSA/Ro60, SSB/La and CENPB (Kappa>0.75), while the coincidence rate foranti-Sm, dsDNA, Nuc and Rib-P detection were moderate (0.4

Objective To investigate the expression of miR-218 (microRNA-218) in breast cancer tissues and its clinical significance. Methods Totally 45 tissue biopsies gained from breast cancer patients and the adjacent normal tissue were collected. Real-time qPCR was used to detect the miR-218 expressions. The correlations of miR-218 expression with clinicopathological characteristics and prognosis of breast cancer patients were analyzed. Results The average expression level of miR-218 in breast cancer tissues was significantly lower than that in control tissues (P = 0.001). The average expression level of miR-218 in PR negative breast cancer tissues was significantly lower than that of PR positive breast cancer tissue (P = 0.037). The average expression level of miR-218 in Ki-67 positive breast cancer tissues was significantly lower than that in Ki-67 negative breast cancer tissue (P=0.018). Conclusion The expression of miR-218 is closely related to carcinogenesis ,progres-sion and prognosis of breast cancer ,so it may be served as a diagnostic biomarker and a prognostic predictor in breast cancer patients,which provides a new way for the treatment of breast cancer.

Objective To investigate the inhibitory effect of methylprednisolone (MP) on the development of coronary arteritis in a murine model of Kawasaki disease (KD) induced with a candida albicaus watersoluble fraction (CAWS).Methods Forty-five C57BIL/6mice were evenly divided into three groups (the control group,the CAWS group and the MP group).Mice in the CAWS group were intraperitoneally injected phosphate buffer saline (PBS) for 5 days.MP or PBS was administered to the different group.The animals were sacrificed at day 3,day 10 and day 28,and the status of vasculitis in the coronary arteries and the aortic root was investigated histologically.One-way analysis of variance (ANOVA) was used to compare the differences among three groups,and t-test for two independent groups.Results The mice in CAWS group and MP group,which induced by CAWS,showed that the body weight and heart weight decreased significantly,and the spleen weight was increased at day 10 and day 28 (P＜0.05).Vasculitis was induced in the mice of those two groups,and the severity score was the highest at day 10 (12.7±0.5).In addition,the severity of the inflammation of the aortic root and the coronary arteries were reduced in MP group (t=6.35,5.55,2.8,P＜0.05).Elastic fiber staining showed that the layers of vascular walls were in disorder and elastic fibers were broken in the CAWS group.However,there was no disruption or breakage in the MP group.Conclusion MP can suppress the progression of coronary arteritis in this CAWS-induced murine vasculitis model,which indicates the efficacy of MP in KD patients with coronary artery lesions.

Objective To investigate the effects of 5-Aza-CdR (methylation transferase inhibitor) on gene expression levels of interleukin-1β (IL-1β),transforming growth factor-β (TGF-β) and nitric oxide synthase (NOS) in chondrocytes.Methods Chondrocytes from patients with osteoarthritis (OA) (n=22),rheumatoid arthritis (RA) (n=3) or trauma without rheumatic diseases (n=10) were collected and cultured.The mRNA expression levels of IL-1β,TGF-β and NOS were detected by real-time polymerase chain reaction (RT-PCR).Chondrocytes in trauma group were treated with 5-Aza-CdR(10μmol/L) for 72 h,and the mRNA and protein expression levels of IL-1β,TGF-β and NOS were detected by RT-PCR and ELISA,respectively.Results The mRNA expression levels of IL-1β,TGF-β and NOS were increased in OA and RA group as compared with trauma group (P＜0.05),while they had no differences between OA and RA groups.After treated with 5-Aza-CdR,the mRNA and protein expression levels of IL-1β,TGF-β and NOS in chondrocytes rised in trauma group as compared with pretreatment (all P＜0.05).Conclusions The mRNA expression levels of IL-1β,TGF-β and NOS in chondroytes are higher in OA and RA patients.5-Aza-CdR could increase the mRNA and protein expression levels of IL-1β,TGF-β and NOS by inducing relative gene methylation,which suggests demethylation might play a role in OA pathogenesis by influncing the inflammatory signal pathway or cell apoptosis.