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 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 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 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 fabricate an anti?tuberculosis controlled drug release coating with Ti?PDA?PEG?PLGA?INH and to investigate its surface characteristics, in vivo and in vitro drug release behavior, and tissue biocompatibility. Methods 4?arm?polyethylene glycol (PEG) was synthesized first. Then cover the surface of titanium (Ti) with a layer of poly dopamine (PDA) by Michael addition reaction. Use porous starch and 4?arm?PEG as a carrier, load with isoniazid (INH), then attach to the surface of titanium by casting or sol?gel dip coating methods, and then cover with a layer of poly lactic?co?glycolic acid (PLGA) by the same method, to fabricate the Ti?PDA?PEG?PLGA?INH composite coating finally. The functional group of 4?arm?PEG was charac?terized by proton nuclear resonance spectroscopy (HNMR). The surface characteristics of Ti?PDA?PEG?PLGA?INH were evaluated by scanning electron microscope (SEM), while drug release behaviors were detected by high performance liquid chromatography (HPLC) and the cumulative release rate was calculated, and carry out the antibacterial performance in vitro. The animal model of femoral condyle bone defect was established in 25 New Zealand white rabbits. Titanium rods covered with PDA?PEG?PLGA?INH coating were implanted into defect area. INH concentrations were detected by HPLC in venous blood, muscle and bone tissue at each time point postoperatively. Another 12 rabbits were randomly divided into experimental group and control group, the experi?mental group was implanted with titanium tablets and titanium rods coated with PDA?PEG?PLGA?INH in the paraspinous muscle and left femoral condyles respectively, while the control group was implanted with a blank sheet of titanium tablets and titanium rods in the same place. Hematoxylin and Eosin Staining were used to observe the biocompatibility of the composite system in vivo at 28 and 56 days postoperatively. Results Ti?PDA?PEG?PLGA?INH controlled drug release coating uniformly distributed on the surface of plates and rods, with translucent form and smooth surface. In vitro INH release kinetics exhibited a short?burst release during the first 8h, and the cumulative release of the INH was about 65%. On the 9th day, the cumulative release of the INH was about 90%, and then the release tended to be flat, and the drug release behavior in vitro continued more than 20d. In vivo release test showed that the concentration of INH in vein blood, muscle and bone tissue around the composite system was increased steadi?ly postoperatively. On about the 28th day, the concentration reached the max. However, the INH concentrations in muscle and bone tissue around the composite system were still higher than the minimum inhibitory concentration (MIC) on the 56th day. The antibacterial test in vitro showed that the titanium tablets coated with PDA?PEG?PLGA?INH formed obvious bacterial inhibition zones. The pathological results indicated that mild inflammatory reaction was seen in the 4th week postoperatively, and the reac?tive capsule formed with loose connective tissue. In the 8th week postoperatively, there's no obvious inflammation occurred, and the reactive capsule became more dense and thicker. Conclusion The study successfully fabricated the Ti?PDA?PEG?PLGA?INH anti?tuberculosis controlled drug release coating, with reasonable release behavior both in vivo and in vitro, effective antibac?terial effect of Mycobacterium tuberculosis in vitro and good tissue biocompatibility, which is a potentially effective drug delivery system for spinal tuberculosis.

Objective:To predict and identify HLA-A * 0201 restricted CD8+ CTL epitopes in Mycobacterium tuberculosis (Mtb) antigen Ag85C, so as to provide evidence for epitope-based study for tuberculosis (TB) vaccine. Methods: The online database SYFPEITHI was applied to predict the potential HLA-A * 0201 restricted epitopes from Ag85C, an antigen of Mycobacterium tuberculosis. T2 cell line was used to assay the affinity between the predicted peptides and HLA-A * 0201 molecules. The specific CTL lines were induced from peripheral blood mononuclear cells (PBMCs) of HLA-A * 0201 positive TB patients and PPD+ healthy donors by peptides with high binding affinity to HLA-A * 0201 molecules. IFN-?production, in vitro proliferation and cytotoxicity of peptide-induced CTL were determined to screen HLA-A * 0201 restricted CD8+ CTL epitopes from those candidates. Results: Fourteen potential epitopes were identified from the SYFPEITHI database. After binding affinity assay, 3 of the 14 peptides (170-178 aa, 317-325 aa, and 144-153 aa) were found to have high binding affinity to HLA-A* 0201 molecules. However, only one peptide (144-153 aa) stimulated its specific CTL to release IFN-y, proliferate in vitro and produce specific cytotoxicity. Conclusion: We have successfully identified a HLA-A * 0201 restricted CD8+ CTL epitope of Mtb Ag85C-FLTREMPAWL( 144-153 aa) , which might be a candidate epitope for TB vaccine designing. Our findings provides a basis for developing novel and effective anti-TB vaccine.

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.