Objective To construct the recombinant plasmid pCMV-Myc-PIASX? and express the fusion protein in mammalian cells.Methods PIASx? fragment was digested from the original vector pGADT7 with SalⅠand NotⅠ,and then was inserted into the targeted pCMV-Myc vector by the recombinant DNA technique.After identification,the recombinant plasmid was transfected into CHO cells.The expression of recombinant Myc-PIASx? fusion protein was detected by Western blotting.Results By the restriction enzyme digestion,fragment purification,ligation and transformation,the recombinant plasmid was obtained.The right recombinant plasmid pCMV-Myc-PIAS3 was identified with enzyme digestion and sequencing.By EcoRⅠ digestion analysis,pCMV-Myc-PIASx? showed a 5641 bp band.By XbaⅠdigestion analysis,pCMV-Myc-PIASx? showed two expected band of 3291 bp and 2349 bp.A specific protein expression band at 68 000(PIASx? fusion protein) was showed in Western blotting,which matched recombinant plasmid.Conclusion The recombinant plasmid of pCMV-Myc-PIASx? is sucssesefully constructed,which provids a good tool for further function study on PIAS family.

Objective To investigate the specimen source and gene phenotype of ESBLs in ESBLs-producing Klebsiella pneumonia of people′s hospital of Sanya city,so as to provide basis for clinical use of drugs and nosocomial infection.Methods Klebsiella pneumoniae was isolated from specimens during January 2013 to December 2014,bacteria identification and susceptibility tests were detected by Phoenix-100 system biochemical,supplementary susceptibility test was confined by K-B method according to 2014 CLSI standards.WHONET 5.6was used in the statistical analysis of all data.Results Totally 213 strains Klebsiella pneumoniae were isolated.The detection rates were 78.4％ of the respiratory secretions,8.92％ and 5.2％ respectively of the secretion and the midstream urine.The strains had a certain resistance to commonly used antimicrobial.The highest resistance rate was 98.1％ to cefotaxime,and the lowest resistance rate was 2.86％ to imipenem.There were 195 in 213 ESBLs producing Klebsiella pneumoniae strain were detect one or more drug resistance gene.The detecting rates of 6 p-lactamase gene of CMY,CTX,TEM,SHV,DHA1 and KPC were 6.10％,76.53％,59.62％,76.06％,12.21％ and 2.82％.Conclusion Klebsiella pneumoniae is mainly isolated from respiratory secretions in the hospital,has a certain resistance to commonly used antimicrobial.We should learn more about the distribution of resistance genes of ESBLs strains,improve the efficiency of the treatment of the infection and to control nosocomial infection and the incidence of multi-drug resistance.

Objective To construct the eukaryotic expression recombinant PIAS3 plasmid fused with Myc protein and express the fusion protein Myc-PIAS3.Methods The full length PIAS3 fragment of 1 851bp was amplified by PCR and ligated into pMD18-T vector. The full length PIAS3 fragment was subcloned into eukaryotic pCMV-Myc vector between SalⅠ and NotⅠ sites.The recombinant pCMV-Myc-PIAS3 plasmid was trandfected into PC3 cells.The eukaryotic expression Myc-PIAS3 protein was checked by Western blotting with Myc antibody.Results The recombinant plasmid showed right sequence by the full length sequencing.The recombinant plasmid of pCMV-Myc-PIAS3 was identified by enzyme digestion.As expected,by EcoRⅠ digestion,it showed two bands of 4 357bp and 1 318 bp. By XbaⅠdigestion,it showed two bands of 3 291 bp and 2 384 bp.The sequencing result showed a N-terminal Myc of 13 amino acids followed with PIAS3 gene sequence in right reading frame.The pCMV-Myc-PIAS3 plasmid was transfected into prostate cancer PC3 cells.A specific protein expression band at relative molecular mass 68 000 was obtained by using Myc-antibody with Western blotting method.Conclusion The recombinant plasmid of pCMV-Myc-PIAS3 is sucssesefully constructed,and Myc-PIAS3 fusion protein is sucssesefully expressed.

Objective To evaluate the levels of serum TAM and Cyfra21-1 in diagnosis and chemotherapeutic efficacy assess-ment of patients with esophageal cancer.Methods The serum TAM and Cyfra21-1 levels were measured in 92 patients with initially diagnosed esophageal cancer or postoperative recurrence of esophageal cancer from September 2012 to September 2013 and 50 cases of healthy people in the same periods as control.The sensitivity and specificity of TAM and Cyfra21-1 were analyzed.Moreover,60 patients with high TAM and Cyfra21-1 levels before chemotherapy from September 2012 to A-pril 2014 were detected TAM and Cyfra21-1 levels after two cycles of chemotherapy.Relation between changes of TAM or Cyfra21-1 and chemotherapeutic efficacy were investigated.Results The diagnostic sensitivity of TAM for esophageal canc-er was 71.7%,which was higher than that of Cyfra21-1 (51.1%,P = 0.004).The diagnostic specificity of TAM and Cy-fra21-1 for esophageal cancer was 94.0% and 92.0% respectively.There were no significantly different between TAM and Cyfra21-1 (P =0.695).Of patients undergoing chemotherapy,the overall response was 25 cases,progress was 11 cases. There was no statistically significant difference in the coincidence rate of TAM and Cyfra21-1 (77.8% vs 75.0%,P =0.781).Conclusion Detection of serum TAM and CYFRA21-1 was valuable in the diagnosis and assessment of the thera-peutic efficacy of chemotherapy in patients with esophageal carcinoma.TAM was better than Cyfra21-1 in the diagnosis of e-sophageal carcinoma.