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 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.

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 the pathogenic bacteria distribution and risk factors of pulmonary infection after tracheotomy in patients with stroke coma,and to put forward preventive measures.Methods:96 patients with stroke coma from January 2016 to February 2017 in our hospital were retrospectively analyzed.The incidence of pulmonary infection and distribution of pathogenic bacteria of patients with stroke coma were analyzed.At the same time,the risk factors of pulmonary infection were analyzed by single factor and multiple factors logistic regression analysis,and corresponding preventive measures were put forward.Results:The incidence of pulmonary infection after tracheotomy in 96 patients with stroke coma was 48.96％ (47/96).A total of 104 pathogens were isolated and cultured,including gram negative bacteria 69 strains (66.35％),gram positive bacteria 20 strains (19.23％) and fungus 15 strains (14.42％).Single factor regression analysis results showed that pulmonary infection after tracheotomy in patients with stroke coma was closely related with age,basic diseases,time of tracheotomy,and bed time,use of broad-spectrum antibiotics,smoking history,artificial airway,times of sputum suction and inhalation(P＜0.05),and it was not related to the patient's gender,weight,stroke type (P＞0.05).Multivariate logistic regression analysis showed that age 45 years old,complicated with basic disease,time oftracheotomy 5 d,use of broad-spectrum antibiotics,smoking history and the establishment of artificial airway were risk factors of pulmonary infection after tmcheotomy in patients with stroke coma (P＜0.05).ROC analysis results showed that the critical point (threshold C) oftmcheotomy time was 4.3 days,and the sensitivity and specificity were 0.851 and 0.918 respectively.Conclusion:The main pathogenic bacteria of pulmonary infection after tracheotomy in patients with stroke coma is gram-negative bacteria,age 45 years old,complicated with basic disease,time of tmcheotomy 5d,use of broad-spectrum antibiotics,smoking history and the establishment of artificial airway can lead to pulmonary infection after tracheotomy in patients with stroke coma,and the risk of pulmonary infection in patients with stroke coma will increase considerably after the time of tracheotomy for more than 4.3 days.Targeted measures should be taken to reduce the risk of pulmonary infection according to pathogenic features and risk factors.

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.

Genetic Diseases, Inborn ; economics ; prevention & control ; therapy ; Humans ; Metabolic Diseases ; economics ; prevention & control ; therapy