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 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.

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.

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 the antitumor mechanism of astragaloside combine with cetuximab on the colon cancer cell line RKO of EGFR over expression through cell proliferation and autophagy. Methods The cell proliferation of colon cancer cell line RKO intervened by astragaloside with or without cetuximab was detected by 4- methyl- teerazolium (MTT). The expressive changes for protein of EGFR 、P62 and LC3 in RKO cells was detected by western bolt. Results MTT showed that 100 ug/mL astragaloside combined with 120 ug/mL cetuximab had significantly inhibiting effect on RKO cells of EGFR expression. Western blot showed that astragaloside can affect the expression of P62 and LC3 to suppress the occurrence ofautophagy. Conclusion In the vitro studies showed that astragalosidecan enhance the antitumor cell proliferation of cetuximab through inhibit autophagy in the treatment of colon cancer.

Objective:To investigate influences of two different HLA-B antigens expressed on K562 cells on receptors expression of NK cells from peripheral blood lymphocytes.Methods:Studied the alteration of the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells before and after PBMC interaction with K562 cells for 24 hours,and also compared the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells after PBMC interaction with two different kind of K562 cells transfected with HLA-B39 and HLA-B51 respectively.Results:After PBMCs were incubated with K562 cells for 24 hours,the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells were both increased.However,after PBMCs were incubated with K562-HLA-B51 cells for 24 hours,the percentage of KIR3DL1+ cells and the percentage of CD16+CD56+ cells were both decreased in comparison with that interaction with K562-HLA-B39 cells.Conclusion:CD16 up-regulation was associated with an up-regulation of inhibitory receptors(KIR3DL1).The interaction between HLA-Bw4 and KIR3DL1 would down-regulate the expression of KIR3DL1.In addition,KIR3DL1 down-regulation was associated with down-regulation of activating receptors(CD16).

Objective To investigate whether MICA gene exon 2,3 and 4 polymorphism is associated with seronegative spondylarthropathies (SpA) or not in Chinese Han population.Methods All 199 B27 positive patients with SpA and 183 randomly ethnically matched healthy controls,and 12 B27 positive controls were enrolled to detect the MICA genotype from its exons 2,3 and 4 by using PCR SSOP method in Shanghai area.Results The MICA007 allele frequency was significantly more in the patient group (18 0%) than in the randomly healthy control group (6 6%) (RR=3 04, P =0 000 045).However,the MICA007 allele frequency was not significantly higher in the B27 positive patient group than in the B27 positive control group.Conclusion The MICA007 allele itself may not be the real disease susceptibility gene involved in the development of ankylosing spondylitis.The increased frequency of MICA007 allele is supposed to be due to a strong linkage disequilibrium between MICA and HLA B genes.

Objective To study the relationship between human leukocyte antigen-DRB1 (HLA-DRB1) allele genes polymorphism and intrahepatic cholestasis of pregnancy (ICP). Methods Forty-two patients with ICP were tested for HLA-DRB1 allele genes polymorphism with the polymerase chain reaction technique and sequence specific oligonucleotide (PCR-SSO) probes hybridization, 56 normal pregnant women as control group were also tested. In addition, the phenotype frequencies of HLA-DRB1 alleles were compared with it′s clinical character in patients with ICP. Results The higher frequencies were observed for alleles DR9, DR12 and DR4 in both groups. DR6 alleles were detected in 14 cases out of 42 patients. Patients with ICP had a significantly higher frequency of the allele DR6 when compared to control group (16.7% vs 3.6%), with a relative risk (RR) as 6.5 (P