The present study was to analyze the epitopes of autoantigen Dsg3 in pemphigus vulgaris (PV), and to elucidate the molecular mechanisms underlying autoimmune diseases. Based on the sequence analysis of autoantigen Dsg3 in Genbank five fragments of Dsg 3 cDNA (E1, E2, E3, E4, and E5) were cloned using RT PCR. Then PCR products were inserted into the expression vector pGEX 2T, and transformed into JM109 strain of E. coli. Dsg3 recombinant proteins were purified from the cell lysates. The epitopes of Dsg3 in PV patients were analyzed with five Dsg3 recombinant proteins by Western blot. The results showed that Dsg3 E1, E2 and E4 recombinant proteins reacted with PV patients′ sera, but not with control or normal sera. The dominant epitopes of Dsg3 in PV patients were different. The above data indicated that the epitopes of Dsg3 were located in E1, E2, and E4, which might play an important role in the pathogenesis of pemphigus vulgaris.

Objective:To analyze the effect of lymphocyte in patients with psoriasis on proliferation of keratinocytes.Methods:Lymphocytes in lesion and peripheral blood were isolated and multiplied,then cultured together with normal keratinocytes. By MTT method,the living cells were counted in the mixed culture.Results:Compared with normal controls, lymphocytes in lesion and peripheral blood of psoriasis both promote the proliferation of keratinocyte(P

Objective:In view of analysis of role of autoantigen Dsg3 in specific T cell response to understand the molecular mechanism of autoimmune diseases.Methods:Based on the sequence analysis of autoantigen Dsg3,the five fractions cDNAs of Dsg3 were cloned and Dsg3-GST fusion proteins induced by IPTG in E.coli.JM109 were purified,then mixed-cultured with T cell from PV patients.The T cell proliferation response were analyzed by MTT.Results:Dsg3 E1,E2 and E4,E5 stimulated T cell from PV patients,not controls.Conclusion:Dsg3 E1,E2 and E4,E5 contained the epitopes relevant to T-B cell interaction,which play an important role in the pathogenesis of pemphigus vulgaris.

Objective To investigate whether the hyper-reactivity of monocytes from the patients with primary biliary cirrhosis (PBC) to LPS and Poly I ∶ C was associated with miR-146a.Methods Peripheral blood mononuclear cells from PBC patients and healthy controls were stimulated with 10 μg/ml of LPS or Poly I ∶ C.The levels of IL-1 β,IL-6,TNF-α and IFN-α in the culture medium were detected by ELISA.The relative expressions of miR-146a,miR-21,let-7e,miR-155 and miR-125b were detected by RT-PCR.The regulatory role of miR-146a on the production of inflammatory factors in LPS or Poly I ∶ C stimulated THP-1 cells was studied by gain-and-loss function assay.Results The up-regulation of miR146a,which was induced by both LPS or Poly I ∶ C,was impaired in the monocytes from PBC patients.The production of LPS or Poly I ∶ C induced inflammatory factor,could be enhanced by miR-146a down-regulation.Conclusion The hyper-reactivity of monocytes from PBC patients to LPS and Poly I ∶ C was associated with miR-146a.

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.

Objective To investigate the application of PBL teaching mode in laboratory diagnosis teaching. Methods 80 clinical medicine undergraduate students were divided into two groups. Tradition lecture was conducted to control group in laboratory diagnosis teaching,while PBL teaching mode PBL teaching mode was carried out to experiment group,and then the mastery of knowledge and appreciation on these two teaching modes was compared. Results The students in the experiment group mastered knowledge better than those in the control group,and the gratification of appreciation on PBL teaching mode was higher than that on the control group. Conclusion The activeness,energeticity,ability of self-study and combination diathesis of the students were improved through PBL teaching. There was good mutual communication between teacher and students and the teaching quality was improved.

Objective To investigate the IL‐17A/F and IL‐22 ,the Th17 related cytokine ,in the serum of cryptococcal meningitis patients and its clinical implications .Methods We collected peripheral blood from 22 cryptococcal meningitis patients and 20 matched healthy controls from February 2011 to February 2013 .Then the peripheral blood mononuclear cells(PBMC)were seperat‐ed with isodensity centrifugation .The RT‐PCR and ELISA were used to detect the mRNA and protein lever of Th17 related cyto‐kine such as IL‐17A ,IL‐17F and IL‐22 .In addition ,the relationship between these cytokines and cerebrospinal fluid characteristics were analyzed .Results The serum concentration of IL‐17A and IL‐22 were much higher than that of healthy controls (P<0 .01) . Their concentration were even higher in patients with poor prognosis factors ,including higher cerebrospinal fluid protein level and cryptococcal antigen titer ,lower cerebrospinal fluid glucose concentration and increased cerebrospinal fluid open pressure .Conclusion IL‐17A and IL‐22 were involved in the pathogenesis of cryptococcal meningitis and may be a potential prognosis biomarker for cryptococcal meningitis.

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 effect of miR-146a on the proliferation and interleukin (IL)-2 production of T helper cells from primary biliary cirrhosis (PBC) patients.Methods MiR-146a in the peripheral blood mononuclear cells (PBMC),monocytes,T helper cells,cytotoxic T cells and B cells from 20 confirmede PBC patients and age/sex matched healthy controls were detected by quantitative PCR.By gainand-loss of function,the miR-146a's effect on anti-CD3/anti-CD28 activated T helper's proliferation and IL-2 production ability were measured by CCK-8 approach and enzyme linked immunosorbent assays (ELISA),respectively.Statistical analysis were carried out by t-test.Results PBMCs (0.46±0.20 vs 1.00±0.26; t=7.47,P＜0.01),T helpers (0.33±0.13 vs 1.00±0.14; t=6.15,P＜0.01) and monocytes (0.56±0.11 vs 1.00±0.11; t=4.97,P＜0.05),but not B cells (0.91±0.06 vs 1.00±0.14; t=0.97,P＞0.05) and cytotoxic T cells (0.98±0.15vs 1.00±0.12; t=0.22; P＞0.05) from PBC patients had lower miR-146a expression level than that of healthy controls.Inducible up expression of miR-146a was observed in PBC patients'T helpers stimulated with antiCD3/anti-CD28 (1.00±0.18 vs 9.12±2.05; t=8.81; P＜0.01).The activated T helpers from PBC patients had higher proliferative ability [PBC:0.35±0.06 (A); healthy controls:0.26±0.04 (A); t=2.83; P＜0.05] and increased IL-2 production [PBC: (685.60±109.19 pg/ml)]; Healthy controls: [(512.20±72.26) pg/ml; t=2.96; P＜0.05 ] than those of healthy controls.For activated T helpers,the proliferation ability,as well as IL-2 production,was enhanced by miR-146a.Conclusion MiR-146a can down regulate the proliferation and IL-2 releasing of activated T helpers.The reduced miR-146a expression enhances IL-2 production and promotes proliferation of T helper of PBC patients,thus,may be involved in the pathogenesis of PBC.