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 investigate the expression level of Gas6 in patients with Immune Thrombocytopenia (ITP) and its clinical significance.Methods The experimental group was peripheral blood samples collected from 35 cases diagnosed with ITP in hematology department of Changhai Hospital in Shanghai from October 2013 to December 2015.Control group was peripheral blood from 35 healthy examined individuals at the same time.After separating plasm from the two group samples,the protein level of Gas6,IFN-α,IL-4,IFN-γ and IL-17 were measured by Enzyme-linked immunosorbent assay (ELISA),comparison of expressional level of the two groups was measured by t test.Pearson correlation analysis was used to decide the relation between Gas6 and cytokines such as IFN-α.Results The expression level of Gas6 in experimental and control groups was 27.28±7.56 ng/ml vs 20.51±5.39 ng/ml (t=4.314,P＜0.000 1);IFN-γ was 221.67±57.64 pg/ml vs 45.32 ±16.79 pg/ml (t=17.38,P＜0.000 1);IL-4 was 113.86±26.48 pg/ml vs 49.87±14.98 pg/ml (t=12.44,P＜0.000 1);IL-17 was 168.96±47.88 pg/ml vs 109.56±28.97 pg/ml (t=6.28,P＜0.000 1);IFN-α was 34.83±8.12 pg/ml vs 29.89 ± 5.76 pg/ml (t=2.936,P=0.004 5),all with statistical differences (P＜0.05).Pearson analysis showed that Gas6 was positively related with IL-17,IL-4,IFN γ (r=0.564,0.486,0.449,P＜0.05) and there was difference statistically,but Gas6 was not correlated with IFN-α.Conclusion Gas6 may participate in the disease formation of ITP through affection on cytokines secreted by Th cell subsets,and it was the potential therapeutic and predicted target for this disease clinically.

Objective To test the expression level of CD69+CD4+CD25-T cells in peripheral blood from patients with autoimmune pancreatitis,and further analyze its clinical significance.Methods Peripheral blood samples from 32 patients with AIP diagnosed in hematological department,Changhai Hospital and 32 health individuals examined at the same time were collected from September 2014 to December 2016,they were classified as experimental and control groups,separately.Peripheral blood mononuclear cells (PBMCs) was acquired by density gradient centrifugation,CD69+ CD4 + CD25-T cells in PBMCs were tested by flow cytometry,and the expression level of cytokines in plasm was by ELISA.The comparison of varies between the two groups was measured by two independent samples' t test.The relationship between the two measurement data was measured by pearson correlation coefficient.Results The expression levels of CD69 + CD4 + CD25-T in experimental and control groups were 10.36％±3.68％ vs 3.99％±1.45％ (t=9.110,P＜0.0001).The expression level of TGF-β was 399.86±121.88 vs 143.87±56.22 pg/ml (t=10.79,P＜0.000 1),both with statistical significance.The levels of CD69+CD4+CD25-T in experimental was positively correlated with TGF-β (r=0.653,P＜0.001) and negatively with IL-4,IFN-γ,IL-2 (r=-0.442,-0.567,-0.351,P＜0.05) and there was statistical significance.Conclusion CD69+CD4 +CD25-T cells might involve the immunopathology of AIP and could be the potential biomarker for clinical diagnosis and therapy.

Objective To explore the change of microRNA let-7b in heat acclimatization/heat stroke rat models and its relation with HSP70,IL-6,TNF-α,TGF-3 and IL-12,and analyze its clinical significance.Methods 60 male rats with almost the same anal temperature,weight,and weeks' age were selected from Laboratory Animal Center in the Second Military Medical University.They were randomly divided into control,heat acclimatization,heat stroke groups averagely.Heat acclimatization/heat stroke rat models were built in hot climate simulated animal tank,blood was collected from cordis apex and serum was separated.Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to test microRNA let-7b in serum,enzyme-linked immunosorbent reaction (ELISA) was to measure the protein level of HSP70,IL-6,TNF-α,TGF-β and IL-12 in serum.Kruskal-Wallis H test was used to analyze quantitative data in the three groups,Spearman rank correlation coefficient was used to reveal relation between two variables in heat stroke group.Results M values of miRNA let-7b in control,heat acclimatization,heat stroke groups were 0.99,1.04 and 1.93 separately,Q values were 0.30,0.25 and 0.44 (x2 =38.95,P＜0.001),separately,with statistical significance.The results of pairwise comparison showed no statistical difference in control and heat acclimatization (P＞0.05),but there were differences in control and heat stroke,heat acclimatization and heat stroke groups statistically (P＜0.05).Spearman correlation analysis showed that in heat stroke group,let-7b was positively related with HSP70,TNF-α and IL-6 (rs =0.579,0.498 and 0.609,P＜0.05) with statistical significance.Conclusion miR let-7b might involve in the pathology of heat stroke,it provided a potential biomarker for monitoring patients in high temperature and humidity clinically.

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 evaluate comparability of two different assay system for detecting CRP.Methods Following the profile of Clinical and Laboratory Standard Institute (CLSI)document EP9-A2,50 blood samples with anti-coagulant ED-TA-2K were collected from emergency patients at Changhai Hospital.The test result of samples by the i-CHROMA Reader was compared and evaluated with those by Beckman Immage 800.Results The linear regression equation for plasma CRP was:Y=1.076 5X-3.031 5,R2=0.986.The linear regression equation for whole blood CRP was:Y=0.882 6X-1.180 8, R2=0.931 1.For whole blood samples with low HCT (<30.45%).Used correction equation:CRP (after corrected)=CRP (before corrected)/(1-HCT).The regression equation (after corrected)was:Y=1.006 8X-3.612 2,R2=0.950 9.Con-clusion CRP concentration detected by i-CHROMA showed good correlation and comparability compared to laboratory ref-erence system by using plasma samples.Results form whole blood samples with low HCT should be corrected to improve comparability.

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 investigate the role of Th9 cells in primary biliary cirrhosis and its clinical significance.Methods The percentages of Th9 cells in patients with PBC (n=38)and healthy controls (n=38)were measured by flow cytometry. Patients clinical data were collected and Mayo risk scores were calculated.Determined the levels of IL-9,PU-1 and TGF-βmRNA expressions and the protein levels of IL-9 in plasma.Further analyzed the correlation between Th9 cells and clinical parameters.Results Compared to healthy controls,Th9 cells percentage was higher in PBC patients and increased with dis-ease stages (t=27.29;P<0.01).IL-9,PU-1 and TGF-βmRNA expressions were increased compared with healthy controls (t=14.69,23.92,10.48;all P<0.01)and the protein level of IL-9 was also up-regulated (t=11.66;P<0.01).The clinical parameters (ALP,AST,ALT,GGT and TBIL)were higher than healthy controls (t=10.94,12.95,10.56,14.92,27.70;all P<0.01).Moreover,the percentage of Th9 cells were positively correlated to Mayo risk,ALT,AST,GGT and TBIL (P<0.05).Conclusion Th9 cells may be involved in the pathogenesis of PBC and related to the disease stage,which provides new clues for future immunotherapy.

Objective To investigate the clinical distribution characteristics of persisters isolated from the chronic infected pa-tients,so as to provide scientific basis for effective clinical measures to prevent,control and treat persister-associated chronic infection.Methods Clinical microbial samples cultured from Jan.2013 to Dec.2014 were analysed by WHONET5.6.Four bacteria species with the highest isolation rate were performed for screening of chronic infection.Concentration of bacteria were detected by viable plate count method and then the growth curve were drew of each sample under the presence of anti-biotics.Persisters were comfirmed according to the specific growth curve under the presence of antibiotics.Results Four highest isolated bacteria species of the clinical samples were Escherichia coli (1 3.2%),Klebsiella pneumoniae (9.6%), Pseudomonasaeruginosa (8%)and Staphylococcusaureus (6.6%).862 chronic infection samples were generated out of 14 216 microbial samples and 41 persisters (4 strains of Escherichia coli,23 strains of Klebsiella pneumoniae,8 strains of Pseudomonasaeruginosa and 6 strains of Staphylococcusaureus)were isolated finally.Conclusion With such a comprehen-sively retrospective analysis of the hospital clinical microbial samples,can tell that the ratio of persisters in chronic infection was not high.And chronic infections are mostly caused by gene-mutated drug-resistant bacteria.However,the isolation rate of Klebsiella pneumoniae was relatively high,of which more attention should be payed to the prevention and control.