Objective The aim of this study was to assess the status of protease-activated receptor-2(PAR-2) expression on the peripheral blood immune cells including dendritic cells(DC) of Wegener's granulo matosis(WG) patients.Methods Flow cytometry was used to analyze the expression of PAR-2 protein on peripheral blood immune cells,DC and DC-like monocytes of healthy controls (HC),inactive,active and different medicine-treated WG patients.Two-tail Mann Whitney non-parametric test was used for statistical analysis.Results There was no difference of PAR-2 expression on PMN,monocytes,lymphocytes and DC between WG patients and HC.However,PAR-2 expression on CD14+CD16+ (t=3.823,P=0.004) and CD64+CD16+ (t=2.652,P=0.024) DC-like monocytes was much higher in WG patients compared with HC.In active WG,expression of PAR-2 was up-regulated on the cell surface of PMN (t=2.690,P=0.013 ),monocytes (t=1.688,P=0.047),lymphocytes(t=1.742,P=0.038),BDCA3+DC(t=2.582,P=0.016),CD11c+DC(t=1.828,P=0.044),CD14+CD16+ (t=3.419,P=0.002)and CD64+CD16+ (t=3.494,P=0.005) DC-like monocytes when compared with inactive WG.PAR-2 expression on these cells was similar between cyclophosphamide (Cyc) and Methotrexate (MTX) treated WG patients.Conclusion PAR-2 expression on immune cells,DC and DC-like monocytes correlates with WG occurrence and development.There is no difference in the effects on PAR-2 expression between various medications.

objecive To chrify clinical characteristics and outcme of acute/subacute interstitial pneumonia(A/SIP)in patients with dermatomyositis.Methods The elinical data of 10 dermatemyositis patients accompanied with A/SIP who hospitalized in April 2006 to April 2008 were reviewed.Data of 9 dermatomyesitis patients with non-A/SIP interstitial lung diseases treated during the same period were also documented for the comparison.The survival rate of patients wag statistically analyzed.Results Compared with those dermatomyositis patients with non-A/SIP interstitial lung diseases,patients with A/SIP had shorter disease courses and higher incidences of fever,heliotrope rash and ground glass opacity in CT image(P＜0.01or＜0.05).However,the levels of serum creatine kinase tended to be normal.After following up 6 months,only 1 patientwithA/SIP survived(P=0.0001).Logistic regression analysis showed the combination treatment of hormonal,cyclophosphamide and cyclosparine might prolong the survival time(P=0.107).Conclusions A/SIP with dermatomyositis is a fatal disease which needs to be early diagnosed and treated.Patients having dyspnea or breathless in the early stage,especially those with recurrent fever,heliotrope rash and normal serum creatine kinase is predicted to develop A/SIP later.A better outcome may be achieved when treating the patients with stemids plus cyclophosphamide and cyclosporine.

Objective To assess the effects of protease 3(PR3)and protease-activated receptor (PAR)-2-activator on the maturation and functions of peripheral blood dendritic cell(DC)-like monocytes.Methods Density gradient centrifugation was used to isolate peripheral blood mononuclear cells(PBMC)from Wegener's granulomatosis(WG)patients and healthy controls(HC).PBMC were stimulated by LPS,human PR3,trypsin,PAR-2-agonist peptide (PAR-2-AP),LPS+PR3 or LPS+trypsin for 24 h.Flow cytometry was used to analyze the expression of PAR-2,CD80,CD83,HLA-DR on stimulated DC-like monocytes-CD14+CD16high monocytes.ELISA kit was used to test the concentration of IL-6 in the culture supernants.Mann-Whitney non-parameteric test was used fur statistical analysis.Resuits No effect of PR3,trypsin and PAR-2-AP on the expression of PAR-2,CD80,CD83,HLA-DR of DC-like monoeytes was found.LPS could significantly induce PAR-2 expression in HC[from(5.8±1.5)%to(24.5±4.5)%,P=0.002]and the expression of CD80,CD83,HLA-DR in HC and WG;PR3,trypsin,PAR-2-AP and LPS could all stimulate the secretion of IL-6.Conclusion PR3 and PAR-2 pathway-activators can not promote PAR-2expression and maturation of DC-tike monocytes,but they can induce the secretion of IL-6.

Objective To analyze the prognostic factors and causes of death of patients with systemic lupus erythematosus (SLE).Methods A database with 319 patients were developed.They were newly diagnosed SLE in the Department of Rheumatology and Immunology,Affiliated Drum Tower Hospital of Nanjing University Medical School from 1999 to 2009.Normal distribution of measurement data was presented using mean±standard deviation.The skewed distribution of data was described by median(interquartile range).Using the rate or proportions,the character of classification data was also stated.Survival rate of SLE patients over time was studied by the Kaplan-Meier method,and prognostic factors were analyzed by COX proportional hazards model.Results The 5 year and 10-year survival rates was 96.2%, 88.7%, respectively Prognostic factors affecting survival included duration from onset to diagnosis, anemia, white blood cells in urine, low serum albumin,low C4 level,abnormal ECG and ultrasound echocardiography, pulmonary arterial hypertension (PAH) and systemic lupus erythematosus disease activity index (SLEDAI). However, PAH,duration from onset to diagnosis, low serum albumin were the independent poor prognostic factors and the relative risk and 95% confidence interval were 2.419 (1.052-5.564), 1.162 (1.043-1.294), 0.924 (0.873-0.978), respectively. Renal failure, pulmonary hypertension and infection were the main causes of death,followed by multiple organ failure and lupus encephalopathy. Conclusion PAH, duration from onset to diagnosis, low serum albumin are the important factors predicting poor prognosis. Early diagnosis, timely treatment of SLE organ damages and preventing complications are the key factors to improve the prognosis of patients with SLE.

ObjectiveTo investigate the role of magnetic resonance imaging (MRI) in treatment efficacy evaluation of polymyositis and dermatomyositis.MethodsFifteen patients with polymyositis and dermatomyositis underwent MRI of thigh were included.Scores of MRI signal intensity of the diseased muscle of every patient were compared before and after treatment and the correlation between serum creatinkinase (CK) level and muscle strength grade were also compared.Correlations between muscle strength grade and MRI score,as well as muscle strength grade and creatinkinase level were analyzed.Comparisons between groups were tested by t test,and the relationship between muscle strength and clinical data was analyzed by Pearson's correlation analysis.ResultsThe signal score of MRI was counted before and after therapy(2.37±0.62,1.30±0.28,respectively,P＜0.05),and CK level[(3841±3175),(549±338) U/L,respectively,P＜0.05] and muscle strength (15.1 ±2.4,18.1 ±0.9,respectively,P＜0.05) were assessed at the same time.Muscle strength grade was associated with signal score of MRI and serum CK level,there was a strong correlation between muscle strength grade and signal score of MRI(r=-0.890,P＜0.05 ).ConclusionMRI may be a useful tool for clinical efficacy evaluation in patients with polymyositis and dermatomyositis.

Objective To explore the clinical efficacy of allogenic mesenchymal stem cells transplantation (MSCT) in patients with refractory polymyositis/dermatomyositis (PM/DM). Methods Bone marrow derived mesenchymal stem cells (BM-MSC) or umbilical cord derived mesenchymal stem cells (UC-MSC)were infused intravenously in 8 PM/DM patients. The clinical manifestations and laboratory parameters,including serum creatin in kinase (CK) and manual muscle test 8 (MMT8) score, were compared before and after MSCT. Staistically andlyzed by paired t-test. Results The eight patients were followed up for six to twelve months after MSCT. The level of serum CK decreased from (1681±430) to (886±248) U/L one week after MSCT (P＜0.05) and further decreased at week 2 (474±61) U/L, 1 month (293±89) U/L, 3 month (202±70) U/L and 6 month (175±46) U/L, respectively (all P＜0.05). MMT8 score increased to 1 month [(67±3) vs (45±14), P＜0.05], 3 month [(64±10) vs (45±14), P＜0.05], 6 month [(64±4) vs (45±14),P＜0.05] after MSCT. The dosage of glucocorticoid steroid were tapered in all patients 2 weeks after MSCT [(18±6) mg vs (34±15) mg, P＜0.05]. Clinical symptoms of interstitial pneumonia of both patients were relieved after MSCT, which was confirmed by the result of high resolution CT (HRCT) of the lung.The skin ulcers tended to be recovered after the transplantation in one DM patient. All patients did not develop transplantation related complications. Conclusion Allogenic MSCT is an effective and safe approach for the treatment of refractory PM/DM. However, extensive follow-up study is needed for long-term benefit evaluation.

Objeefive To explore the clinical efficacy and safety of allogenic bone marrow derived mesenchymal stem cells transplantation(MSCT) in patients with refractory systemic lupus erythematosus(SLE).Methods Eleven patients with refractory SLE(nine females and two males aged from 16～41 years(mean 25±8).were entailed in the study.The infcIrmed consents which were approved by the Ethics Committee of the Affiliated Drum Tower Hospital of Naniing University Medical School were obtained from all participants.Bone marrow of healthy donors were obtained and the mesenchymal stem cells(MSC)were expanded in vitro.Each patient was infused MSC 1×106/kg body weight intravenously.Before MSCT.all patients were administrated with cvclophosphamide (CTX)800～1800 mg divided by two to three days.The clinical manifestations and laboratory tests were compared before and after MSCT.Results The eleven patients were followed up for one to thirteen months after MSCT.A11 patients did not develop transplantation related complications.The systemic lupus erythematosus disease activity index (SLEDAI)score decreased from 1 1.7±5.1 to 5.6±3.4 one month after MSCT(n=11,P＜0.01).Ufine protein excretion decreased from(1989+842)mg/24 h to(1118±700)mg/24 h one month after MSCT(n=10,P=0.02).Five patients were followed up for six months and their urine protein excretion decreased significantly [(522±151)mg/24 h vs (2478±797)rag/24 h.n=5.P＜0.01j.The serum albumin level of 5 patients with hypoalbuminemia increased gradually one month after MSCT [(28±6)g/L vs (32±7)g/L,n=5,P＜0.05].Serum complement C3 level increased from(0.50±0.12) g/L to(0.75±0.10)g/L (n=9.P＜0.01) and their anti-nuclear antibody (ANA) titer decreased one month after MSCT.In two patients with chronic renal failure.the serum creatinine decreased gradually.Conclusion Allogenic MSCT is an effective and safe approach for the treatment of refractory SLE.However.extensive follow-up study is needed for long-term benefit evaluation.

Objective To explore the clinical efficacy and safety of umbilical cord derived mesenchymal stem cells transplantation(UC-MSCT)for patients with refractory systemic lupus erythematosus (SLE).Methods Twelve patients with refractory SLE were enrolled in this study.UC-MSCs(≥106/kg cell number)were infused intravenously for each patient. The clinical manifestations and laboratory parameters were compared before and after MSCT. Results The twelve patients were followed up for one to twenty-six months after MSCT.The systemic lupus erythematosus disease activity index(SLEDAI)score decreased from 18±4 to 10±4 one month after MSCT(n=12,P＜0.01)and then decreased to 7±4 at three month follow-up.Nine patients showed improvement of 24 h proteinuria[(2103±749)mg vs(3359±1248)mg,P＜0.01]one month after MSCT.Further improvement of 24 h proteinuria was observed in eight patients[(1427±616)mg vs(3342±1333)mg,P＜0.01]at three months post MSCT.Serum creatinine of five patients decreased significantly and ten patients showed an increase of serum albumin. Serum complement C3 increased in three patients and four patients showed obvious amelioration of hematological abnormalities. There was no transplantation related complications for all the patients. Conclusion UC-MSCT is effective and safe for refractory SLE,but further observation is required to evaluate its long term efficacy.

Objective To investigate the association of human leucocyte antigen (HLA-DRB1) with anti-melanoma differentiation-associated gene 5 (MDA5) expression in polymyositis/dermatomyositis (PM/DM).Methods Seventy patients with PM,104 patients with DM and 400 healthy controls were included.Genotyping of HLA-DRB1 was performed using the sequencing-based typing method.Levels of anti-MDA5 were measued by enzyme linked immunosorbent assay using recombinant MDA5 antigen.The frequencies of HLA-DRB1 alleles were compared between the patients and controls using a chi-square test or Fisher's exact test.Results Frequencies of DRB1 * 04∶01 [17.0％ vs 1.3％,corrected P-value (Pc)=3.8×10-8;odds ratio (OR)=16.2;95％ confidence interval (CI) (6.6,39.7)] and DRB1 * 12∶02 [(42.6％ vs 19.3％,Pc=0.008;OR=3.1;95％ CI(1.7,5.7)] were significantly higher in anti-MDA5 positive PM/DM patients compared with the controls.The frequencies of DRB1 * 04∶01 [P=5.2×10-6,OR=17.1,95％CI:(5.3,54.9)\ and * 12∶02 [P=3.8×10-4,OR=3.1,95％CI(1.7,5.7)] in anti-MDA5 positive DM-interstitial lung disease (ILD) patients were higher than those in the controls,whereas the frequencies of DRB1 * 04∶01 and * 12∶02 did not differ between the anti-MDA5 negative DM-ILD patients and the controls.No difference in the frequency of DRB1 alleles,other than * 04∶01,carrying the shared epitope (SE),i.e.* 01∶01,* 01∶02,* 04∶05 and * 10∶01,was observed between the controls and DM patients stratified by the presence of anti-MDA5 and ILD.Conclusion DRB1 * 04∶01 and * 12∶02 confer susceptibility to anti-MDA5 antibody production in DM,which cannot be explained by the SE hypothesis.

Objective To investigate the expression of soluble vascular endothelial growth factor receptor 1 (sFlt1) in human umbilical cord-derived mesenchymal stem cells (HUCMSC) under the condition of inflammatory cytokine and co-culture with rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs).Methods ① HUCMSC were cultured alone or stimulated by Tumour necrosis factor (TNF)-αt,interferon (IFN)-γor the combination of TNF-α and IFN-γ.The sFh1 levels of each group were detected by enzyme-linked immunosorbent assay (ELISA) at 24 h,48 h and 72 h respectively.② RA-FLSs and HUCMSC were cocultured in a transwell system with both of them cultured alone as control for 72 hours.ELISA was used to detect sFlt1 levels in each group.T test was used for comparison between two groups,and ANOVA was used to compare multi-group variables.Results ① The sFlt1 levels of TNF-α + IFN-γ group were significantly higher than those of the control group at 24 h and 48 h (24 h (5.4±0.4) ng/ml vs (2.8±1.7) ng/ml,t=0.942,P=0.026;48 h (7.2±0.8) vs (4.3±1.0) ng/ml,t=4.285,P=0.005),while that at 72 h was not significantly different [(9.1±1.7) ng/ml vs (7.0±1.4) ng/ml,t=0.683,P=0.52].And no significant difference in sFlt1 levels between control group and TNF-α group or IFN-γ stimulation group were observed.② Compared with RA-FLSs,the expression of sFlt1 in HUCMSC was significantly higher (8.19±3.64) ng/ml vs (0.19±0.08) ng/ml,t=7.280,P＜0.01].After co-cultured with RA-FLSs and HUCMSC,the sFlt1 concentration of the co-culture group was significantly higher than that of HUCMSC group [(17.26±6.92) ng/ml vs (8.19±3.64) ng/ml,t=3.985,P=0.000 7].Conclusion The sFlt1 expression of HUCMSC is up-regulated by inflammatory cytokine TNF-α combined with IFN-γ.RA FLSs may promote the co-cultured HUCMSC to increase sFlt1 expression by secreting inflammatory factors.