Objective:To explore the influence of chaperone tapasin on HLA E expression on cell surface.Methods:The infection method was used to transfer the recombinant retrovirus expression vector which was constructed with HLA E molecules on target cells was detected with FACS technique.Results:Exogenous HLA E expression on T2 cell surface was detected with 74.13% and no HLA E expression was detected on control cells and that was transfected with empty vector.Conclusion:It is possible that the expression of HLA E molecules on cell surface involves in TAP independent mechanism.

Objective:To construct mammalian cell expression vector of human leukocyte antigen C gene and express it in JAR cell and study its functions.Methods:Total cell RNA was extracted from peripheral blood mononuclear cells and the cDNA was amplified by RT PCR;the cDNA fragments were directly inserted into Pbluescript KS(+/-) and the recombinant PBS HLA C was identified by restriction endonuclease digestion and sequencing;then the plasmid of pcDNA HLA C was transfected into JAR cells.Lactate Dehydrogenase(LDH) release assay was employed to detect the cytotoxicities of NK cells proliferated in vitro against HLA C transfectants JAR cells.Results:The recombinant mammalian cell expression vectors of pcDNA3 HLA C was constructed,the sequences of the insert was identical to the published sequences encoding HLA Cw *0602 gene.Several transfected JAR clones were obtained,which expressed HLA Cw *0602 in high level and constitutively.The specific lysis was detected for NK cells stimulated by IL 2 against JAR cells and HLA C transfectants JAR cells.But both cells were resistant to freshly isolated PBL cells.Conclusion:The recombinant mammalian cell expression vectors of pcDNA3 HLA Cw *0602 was successfully constructed,JAR cells resistance to NK lysis could involve an HLA C independent mechanism.

Objective:To investigate the mechanism of association between Pregnancy induced Hypertension and HLA G.Methods:The distribution and frequencies of HLA G alleles were analyzed in 54 PIH patients and 146 random healthy controls with nonradioactive digoxigenin labeled PCR SSO method.Results:The results showed that the same nine HLA G alleles were detected and the G*01011 was the commonest allele with the frequency of 31.38% and 37.33% respectively in both PIH and controls groups,the next was G*01013,25.00% and 20.21% separately.Statistic analysis showed that the frequencies of HLA G alleles were similar to in the two groups and there was no significant difference between PIH patients and controls.Conclusion:These data suggest that there is no significant detectable relation between the HLA G polymorphism and PIH.

Objective:To investigate the effects of recombinant human interluekin-1 receptor antogonist (rhIL-1Ra) on the cartilage repair in rat temporomandibular joint(TMJ) with osteoarthritis(OA).Methods:Collagenase-Ⅱ was injected into bilateral TMJs of 24 adult rats for the induction of bilateral TMJOA,1 week after injection,5μg rhIL-1Ra(diluted in 0.05 ml normal saline) was injected into each right TMJ and the left joint received the same amount of normal saline injection as the control.12 animals were sacrificed at 2and 4 weeks after the first injection respectively.HE staining,immunnohistochemical method and RT-PCR examination were conducted.Mankins scere was used to evaluate the TMJOA degree.1 adult SD rat was used as healthy control,and sacrificed at 2 weeks of the experiment.Results:The TMJs of both sides showed typical OA-related cartilage degradation 2 week after IL-1Ra treatment,the Mankin~ score of the IL-1Ra treated and control joints was 1.33±0.52 and 2.00±6.63 (P＞0.05),4 week after treatment that was 3.00± 0.63 and 6.50 ± 0.84 (P＜0.05),respectively.Lower expression of ADAMTS-4 and ADAMTS-5 was observed in the treated joints than in the controls (P＜0.05).Conclusion:Intra-articular injection of IL-1Ra into TMJ can alleviate the cartilage lesion,the mechanism may lie in the inhibition of the expression of ADAMTS-4 and ADAMTS-5.

Objective To investigate serum nesfatin-1 levels in type 2 diabetes (T2DM)patients with or without nonalcohol-ic fatty liver(NAFLD).Methods The hight,weight,serum GLU,lipid profiles,INS,HbA1c,ALT,nesfatin-1 levels were measured in 21 1 T2DM patients[98 without NAFLD(TD2M without NAFLD group)and 1 13 with NAFLD(TD2M with NAFLD group)] and 75 normal subjects(NGT group).BMI、HOMA-IR were also calculated.Results The serum nesfatin-1 level was significantly higher in T2DM group than those in NGT group(P <0.05),and T2DM with NAFLD group had higher nesfatin-1 level than T2DM without NAFLD group (P < 0.05 ).There was a close relation between nefastin-1 of T2DM group and course of disease (r=-0.447,P <0.05).NAFLD existed a significant correlation with BMI、TG and nesfatin-1 (P <0.05).Conclusion nesfatin-1 may partially contribute to the pathogenesis of T2DM and NAFLD.

PurposeTo study the changes of serum nitric oxide(NO)and endothelin(ET)levels by L-Arg therapy in acute hypoxic model. MethodsNADPH diaphorase histochemical staining method, Griess biochemical assay and radioimmune assay were applied to investigate the changes of nitric oxide synthase (NOS) ,NO, cGMP and ET in normal, hypoxic and L-Arg treated hypoxic rats. Resultsduring acute hypoxia, while serum ET rose up significantly, both the levels of serum NO and cGMP and the activity of NOS were significantly lower. After the application of L-Arg therapy, the activity of NOS did not change significantly, but the levels of serum NO and cGMP rose up significantly, and the level of serum ET was to the opposite direction. ConclusionsNO and ET may modulate hypoxic pulmonary hypertension, and acute hypoxia can result in acute hypoxic pulmonary hypertension. L-Arg can reverse the acute hypoxic hypertension.

ObjectiveTo determine optimal positive end expiratory pressure (PEEP) in mechanical ventilation in children with severe asthmatoid disease based on the quasistatic pressure-volume (P-V) curve.MethodsA serf-control study was done on 23 children with severe asthmatoid disease in the pediatric intensive care unit( PICU ).Quasistatic lung P-V curve of these patients was analyzed and the lower inflection point (LIP) from P-V curve was determined.Three different PEEP (0 cm H2O,LIP,LIP+2 cm H2O,1 mm H2O =0.098 kPa) were given to the patients.The effects of PEEP at different levels on gas exchange,hemodynamic and airway pressure were observed.ResultsThe quasistatic LIP were (2.70 ±2.00)cm H2O.When PEEP was increased to the level of LIP + 2 cm H2O,PaO2 / FiO2 and lung compliance improved significantly (P ＜ 0.01 ) and dynamic lung compliance was the highest,peak inspiratory pressure was (22.30 ± 3.00) cm H2O and mean airway pressure was( 14.11 ± 1.01 ) cm H2O,without obvious adverse effects on mean arterial blood pressure and heart rate.There was no difference in PaCO2,when compared PEEP =0 cmH2O to PEEP =LIP + 2 cmH2O.ConclusionThe application of PEEP is safe.LIP + 2 cm H2O from quasistatic P-V curve could be set as the optimal PEEP under which mechanical ventilation has the best efficacy and do not aggravate CO2 retention and abnormality of hemodynamics in children with severe asthmatoid disease.

ObjectiveTo observe the changes of amplitude-integrated electroencephalogram (aEEG)of neonatal asphyxia with different Apgar scores,and to investigate the diagnostic value of aEEG for hypoxicischemic encephalopathy(HIE) in neonatal asphyxia.MethodsaEEG monitoring were detected on 56 fullterm asphyxia neonates who were hospitalized in our neonatal department from Dec 2010 to Oct 2011.According to 1 minute Apgar score after birth,56 cases were divided into two groups:observation group in which 36 cases with Apgar score 0～7,and control group in which 20 cases with Apgar score 8～10.aEEG monitoring was done within 6 h,2 d,3 d,7 d of each neonates after birth,and the changes of aEEG were analyzed and the diagnostic value on HIE were evaluated.ResultsAmong 20 cases in the control group,the aEEG results in 6 hours after birth were 17 cases (85％) had normal aEEG results,3 cases( 15％ ) mildly abnormal,nd no one severely abnormal.The aEEG results of patients in observation group(36 cases) were 18 cases(50％ ) had normal aEEG results,13 cases(36.1％ ) mildly abnormal,and 5 cases( 13.9％ ) severely abnormal.The abnormal rate in observation group was significantly higher than that of the control group ( x2=5.3,P＜0.001 ).There were 34 HIE patients in the total 36 cases of observation group,whose aEEG monitoring results in 6 hours after birth were associated with HIE clinical grading( Spearman's rank correlation coefficient was 0.867,P＜0.01 ).Dynamic aEEG monitoring for 56 patients showed that 21 cases had abnormal aEEG in 6 hours after birth,in whom 15 cases(71.4％ ) could returned to normal after 48～72 hours after birth,and there were only 4 case (7.1％ ) still had severely abnormal aEEG results in the seventh day after birth.ConclusionThe aEEG dynamic monitoring for full-term HIE neonates after birth enhances early prediction of HIE.

Objective To investigate whether the killer cell immunoglobulin like receptor (KIR) gene polymorphisms are associated with seronegative spondylarthropathies.Methods All 197 HLA B27 positive patients with seronegative spondyloarthro pathies (SpA) and 83 randomly ethnically matched healthy controls were enrolled to detect the KIR genotype using PCR SSP in Shanghai area.Results The KIR3DL1 gene frequency was significantly lower in the patient group (0 763) than in the control group (1 00) (RR=0 76, P =0 003).Meanwhile,the gene frequencies of two pseudogenes (KIR2DP1,KIR3DP1) were significantly higher in SpA group than in healthy control group (RR=1 1, P =0 004).Conclusion There may be a negative association between pathogenesis of SpA and KIR3DL1 gene.The function of KIR3DL1 molecule should be investigated deeply.

Objective:To explore the mechanism of association between recurrent spontaneous abortion(RSA) and the polymorphism of HLA E,had detected the frequencies of five HLA E alleles in 53 women with RSA and 156 random Shanghai Chinese controls.Methods:PCR sequence specific oligonucleotide (PCR SSO) typing was carried to detect polymorphism in exon2 and exon3 of HLA E gene.Results:HLA E *0101 is the most common allele in both RSA patients and controls(50% and 43.09% respectively).HLA E *01031 and E *01032 were detected with allele freqencies of 23.79% and 33.12% in controls,20.75% and 29.25% in RSA.No HLA E *0102 and E *0104 alleles could be detected in both groups.Conclusion:Maternal HLA E polymorphism does not associated with pathogenesis of RSA.