Objective To explore the clinical significance of STING in peripheral blood in patients with primary biliary cirrho-sis.Methods Admitted 28 PBC patients hospitalized in Shanghai Changhai Hospital and Shanghai Changzheng Hospital from January 2014 to October 2015.Also enrolled 28 healthy controls.Baseline data and laboratory indicators were extrac-ted,and STING mRNA expression was determined using q-PCR.The correlation between STING and clinical parameters were analyzed.The changes of STING mRNA in 5 followed-up patients with PBC were analyzed.Results Compared with healthy controls,STING mRNA was significantly increased in PBC patients and was increased in patients with severer dis-ease stages (U=0.00,P<0.05).STING was positively correlated with Mayo risk and GGT (R=0.45,R=0.42,P<0.05),and not AST,ALT or ALP (R=0.33,0.21,0.27,P>0.05).After therapy,STING mRNA were significantly re-duced in 5 PBC patients (U=0.00,P<0.05).Conclusion STING may be involved in PBC pathogenesis.

Objective To investigate the clinical characteristics of twice-weekly hemodialysis patients.Methods Data were collected from Shanghai Renal Registry.A total of 1288 patients undergoing regular hemodialysis (HD) with dialysis adequacy index and other biochemical parameters in Shanghai in January 2007 were enrolled into the cohort study with 2 years follow-up.Clinical characteristics and outcome of twice-weekly HD patients were analyzed as compared with thrice-weekly HD patients.Results Compared with patients on thrice-weekly HD,the twice-weekly HD patients were significantly younger and had significantly shorter HD vintage,smaller body surface area,longer HD session time,higher single-pool Kt/V (spKt/V) and serum albumin but lower weekly Kt/V (P＜0.05).There was no statistical difference in ultrafiltration volume between two groups.Kaplan-Meier survival analysis indicated that both groups had similar two-year survival.Multivariate Cox regression analysis showed that age,body mass index,serum albumin and weekly Kt/V were predictors of patient mortality.Conclusion It is acceptable for some hemodialys patients with twice-weekly HD,and close monitor of dialysis adequacy and volume status is necessary for this therapy model.

Objective To investigate the association between IL-22 and the pathogenesis of coronary artery atherosclerosis(AS).Methods The relative expression of IL-22 mRNA in PBMC from 30 AS patients and 8 patients without any signs of coronary artery stenosis was detected by RT-PCR.Serum IL-22 levels of 22 patients without any signs of coronary artery stenosis and 79 AS patients were detected by ELISA.CRL-1730 cells(human umbilical vein endothelial cells) were stimulated with oxidized low density lipoprotein (ox-LDL) at different dosage for 24 h,and the expression of IL-22R1 was detected by flowcytometry.The proliferation ability of CRL-1730 cells treated with IL-22(20 ng/ml) and/or ox-LDL(100 μg/ml)was measured by MTS assay,and the expression of basic fibroblast growth factor(bFGF) was detected by RTPCR and ELISA.Results Decreased IL-22 expression in PBMC and serum was observed as worsen of AS.The expression of IL-22R1 in ox-LDL treated CRL-1730 cells was increased in dose dependent manner.OxLDL decreased proliferation ability,as well as bFGF expression and releasing,of CRL-1730 cells.This effect of ox-LDL was partially rescued by IL-22.Conclusion IL-22 may have anti-atherosclerosis effect.This effect may be mediated by regulating bFGF expression and endothelial cells proliferation ability in the presence of IL-22.

Objective To explore the clinical significance of neutrophils/lymphocyte(NLR)and platelet/lymphocyte ratioof (PLR)peripheral blood in patients with primary biliary cirrhosis.Methods Retrospectively analyzed 62 patients with PBC and three subgroup patients according to Child-pugh,who were hospitalized in Shanghai Changhai Hospital and Shanghai Changzheng Hospital from January 2010 to July 2015.Enrolled 59 healthy controls.Baseline data and laboratory indicators were extracted,and the correlation between PLR and NLR and disease activity were analyzed.The changes of NLR in 3 pa-tients with PBC were analyzed.3 patients were followed up and their NLRs were recorded before and after therapy.Results Compared with healthy controls,NLR was significantly increased in PBC patients (P<0.05),and not PLR (P>0.05). NLR was positively correlated with TBIL and Mayo risk (R=0.35,0.30,P<0.05),and not ALT,AST or ALP (P>0.05).After therapy,NLRs were significantly reduced in three PBC patients (P<0.05).Conclusion NLR may be a useful biomarker for assessing clinical therapy in PBC patients.

Objective To explore the expression of T cell immunoglobulin and mucin-3(TIM-3)in peripheral blood mononu-clear cells from patients with primary Sj¨ogren’s syndrome(pSS)and its clinical significance.Methods Enrolled 33 pSS pa-tients from Shanghai Changzheng Hospital and Changhai Hospital in October 2012 and July 2013.The relative expression of TIM-3 in the peripheral blood mononuclear cells(PBMC)in 33 patients and 33 healthy individuals was detected by RT-PCR. The association between TIM-3 mRNA level and clinical characteristics were analyzed,for example,serum RF,CRP and anti-SSA/SSB antibody.1 2 of the pSS patients were followed up and their TIM-3 mRNA level was determined after glucocorti-coid treatment for 2 months.Results The expression of TIM-3 in PBMCs from patients with pSS was increased significantly than that in healthy individuals (0.55±0.12 vs 0.13±0.10,t=15.44,P<0.000 1),but its expression between anti-SSA/anti-SSB positive and negative patients were not statistical difference (0.45±0.21 vs 0.54±0.31;0.46±0.15 vs 0.51± 0.24).The TIM-3 mRNA expression in pSS patients was positively correlated with serum RF and CRP (R=0.558 5,R=0.414 7,P>0.05),but was not associated the anti-SSA/SSB antibody (P=0.298 2).TIM-3 mRNA level was decreased in the patients who were treated with glucocorticoids for 2 months (0.62±0.10 vs 0.38±0.13,t=5.07,P<0.05).Conclusion TIM-3 may play a role in the pathogenesis of pSS and it may be regarded as a biomarker to pSS.

Objective To explore TRAIL expression in patients with primary immune thrombocytopenia (ITP)and its role in the pathogenesis.Methods Peripheral Blood Mononuclear Cells (PBMCs)from twenty-eight patients with ITP and thirty healthy controls were obtained.TRAIL expression was determined using Real-time Quantitative Polymerase Chain Reaction (RT-PCR).Serum TNF-α,IFN-γ,IL-4 and IL-10 were detected using ELISA.Associations between TRAIL expression and clinical parameters were assessed.Results Compared to healthy controls,TRAIL expression was significantly increased in PBMC from patients with ITP (2.80±0.43 vs 1.00±0.24,t=19.72,P<0.001).Compared to the healthy controls,ex-pression of IFN-γand TNF-αfrom ITP patients were increased (52.43±11.04 pg/ml vs 23.26±8.15 pg/ml;69.14± 10.89 pg/ml vs 36.14±13.17 pg/ml,t=10.36~11.50,P<0.001)and expression of IL-10 were decreased (11.18±6.13 pg/ml vs 33.28±9.85 pg/ml,t=10.17,P<0.001)and there was no significant difference in IL-4 expression (35.75± 12.52 pg/ml vs 40.16±14.26 pg/ml,t=1.25,P>0.05).Furthermore,TRAIL expression was positively correlated with serum IFN-γ,TNF-αlevel (r=0.432,0.541,P<0.05),and negatively correlated with IL-10 and PLT (r=-0.424,-0.553,P<0.05).Conclusion Rur results suggest that TRAIL may be involved in the pathogenesis of ITP.

Objective To investigate the clinical significance of neutrophils/lymphocyte ratio (NLR)and platelet/lymphocyte ratio (PLR)of peripheral blood in patients with myasthenia gravis (MG).Methods 54 patients with MG treating from Jan-uary 2013 to December 2013 in Changhai hospital and 57 healthy adults who received check-up were enrolled the study as MG group and control group.WBC count,NLR and PLR were compared between two groups and among different clinical classifications in MG patients.The area under the receiver operating characteristic (ROC)curve was performed to evaluate these indicators’diagnostic value for MG.Results The WBC count,NLR and PLR in MG group were (6.85±0.37)×109/L,2.48±0.19 and 118.79±6.38 respectively,which were significantly higher than (5.87±0.12)×109/L,1.59±0.06 and 102.01±3.45 in control group (P <0.05).The area under the ROC curve of WBC count,NLR and PLR were 0.559,0.717 and 0.581 respectively.PLR of MG patients with type Ⅲ or Ⅳ was significantly higher than that of patients with typeⅠ,Ⅱ(P <0.05).Conclusion NLR and PLR of MG patients increased significantly.The diagnostic value of NLR is higher,and PLR may be associated with the severity of disease.

Objective To test the expression of MIF in peripheral blood mononuclear cells (PBMCs)from patients with Cry-tococcal Meningitis and further discuss its clinical significance.Methods Peripheral blood from 42 patients with Crytococcal Meningitis diagnosed in Changhai Hospital,Shanghai and 42 healthy individuals examined at the same time was collected from August,2012 to November,2015.PBMCs were separated by density gradient centrifugation method,mRNA relative expression of MIF in PBMCs was measured by PCR,the level of MIF,IL-17,IL-1β,TNF-α,IFN-γand IL-4 in plasma was tested by Enzyme-linked immunosorbent assay (ELISA).The comparison for expression level of cytokines between the two groups was by two-independent samples t test.Pearson correlation coefficient was used to measure the relation between MIF and other cytokines.Results The protein levels of MIF in experimental and controlled groups were 34.17±7.88 ng/ml vs 10.89±2.76 ng/ml(t=18.07,P<0.0001),while relative expression of RNA was 2.87±0.94 vs 1.95±0.89(t=4.606,P<0.0001),and there was statistical significance (P<0.005).Pearson correlation analysis showed that MIF was positively related with IL-1β,IL-17 (r=0.467,0.401,P<0.01),with statisticaldifference.Conclusion MIF may involve in the im-mune regulation for Crytococcal Meningitis by affecting the secretion and function of cytokines as IL-1β,IL-17,and it was potential target and monitored biomarker for this disease.

Objective To test the expression level of IL-37 in peripheral blood mononuclear cells (PBMCs)of patients with primary biliary cirrhosis (PBC)and further explore its clinical significance in the pathological process of PBC.Methods Pe-ripheral blood samples were collected from 42 patients diagnosed as PBC and 38 health individuals examined at the same time during June 2013 to August 2015 in Changhai Hospital.PBMCs were separated by sucrose density gradient centrifugation, qualified Real Time-Polymerase Chain Reaction (qRT-PCR)was used to measure IL-37 mRNA expression level in PBMCs. Enzyme-Linked Immuno Sorbent Assay (ELISA)was to measure the protein level of IL-37,IL-6,IL-17,TNF-α,TGF-β,IL-18 and IL-23 in plasma.Meanwhile,the pathological stages of PBC cases were recorded.Pearson correlation analysis was performed on IL-37 and IL-6,TNF-α,IL-17,TGF-β,IL-18 and IL-23.Spearman rank correlation analysis was on IL-37 and pathological stages of PBC.Results The mRNA and protein level of IL-37 in experimental and controlled group were 2.81 ±0.94 vs 1.09±0.56,356.14±169.36 pg/ml vs 86.68±48.23 pg/ml separately(t=9.811,9.462,P<0.000 1),with sta-tistical differences.The correlation analysis showed that IL-37 was positively related with IL-17,TNF-α,IL-6 and TGF-β(r=0.561 2,0.661 9,0.672 1,0.765 3,P<0.001),and disease stages (Ⅰ~Ⅳ)(rs=0.348 9,P<0.05).Conclusion IL-37 might involve in the pathological process of PBC,and it is significant for disease prediction and diagnosis.

Objective To study the activation induced cell death (AICD) of CD4+ T cells in primary biliary cirrhosis(PBC)murine model induced by poly I∶C. Methods Thirty female C57BL/6 mice were divided into model and control group randomly, and the former were injected with 5 mg/kg of poly I∶C, the later with PBS. PBC mice were detected 16 weeks after injection. CD4+ T cells isolated from spleen were stimulated in vitro by Con A and anti-CD3, and the apoptosis were determined by Annexin-V and PI staining. The expression of Fas, FasL and TRAIL were assayed by relative quantitative real-time PCR. Bcl-2 was detected by Western blot. Results Compared with control group, the portal areas of mice in model group were infiltrated with mononuclear cells obviously. The positive rate of serum antimitochondrial antibody(AMA) and the level of alkali phosphatase (ALP) were higher than that in control group (P<0.001). AICD of splenic CD4+ T cells in model group was lower than that of control group (P<0.001). The mRNA of FasL and TRAIL in model mice was down-regulated. Simultaneously, the anti-apoptosis protein Bcl-2 was up-regulated in model group. Conclusion These observations suggest that a defect in AICD of auto-reactive TH1 cells may contribute to the pathogenesis of PBC model. Furthermore, this defect in AICD may results from the change of Fas/FasL, TRAIL pathway and the up-regulation of Bcl-2.