Objective To investigate the production of carbapenemase and 16S rRNA methylase in five isolates of pan-drugs resistant E.cloacae recovered in Ruijin hospital.Methods MICs of the five isolates to 10 antibiotics were determined by E test.Six kinds of 16S rRNA methylase genes and a series of β- lactamase genes were amplified by PCR.Shotgun cloning was performed to detect carbapenem resistance determinant.The conjugal transfer of carbapenemase gene and 16S rRNA methylase gene was performed in broth culture with E.coli J53 as the recipient.Pulsed-field gel electrophoresis (PFGE) was carried out to analyse the genotyping.IEF was performed to detect β-1actamases.Southern blot was performed to determine the location of carbapenem resistance determinant.Results The MICs of 10 antibiotics were ＞32 mg/L.Four β-1actamases with pIs of 5.4 ( TEM-1 ),6.7 ( KPC-2 ),8.2 ( SHV-12 ),8.4 (CTX-M-14) were determined.The insertion sequence in the recombinant plasmid was blaKPC-2 flanked by a transposon.blaKPC-2 was located on a large non-conjugative plasmid whereas armA was located on an other conjugative plasmid.PFGE patterns of 5 isolates were identical.Conclusion KPC-2 was responsible for carbapenem resistance in pandrugs resistant Enterobacter cloacae.There was no relationship between blaKPC-2 and armA.Although pandrug resistant Enterobacteriaceae remain rare,the emergence of this group of organism merits monitoring.

AIM:To study the differentiation of rat marrow-derived mesenchymal stem cells(MSCs) into myoid cells in vitro . METHODS:The MSCs of SD rat were cultured?passaged?induced and differentiated in vitro used by routine culture technique, and evaluated by FACScan flow cytometer,detected by immunohistochemistry and analyzed by Hitachi H-600 transmission electron microscope(TEM). RESULTS:FACScan shows that cells expressed the antigens of CD29 and CD44, not those of CD11b and CD45; cells show positive response of staining with desmin and myoglobin after processing by two compounds of 5-azacytidine and amphotericin B. There were stripform zone of myofilament without any organells beside the edge of membrane in myoid cell of induction and differentiation checked by TEM. CONCLUSION: The passaged cells were MSCs and the MSCs may have specific gene structures that can differentiate into myoid cells. The demethylation or hypomethylation may conduct by compounds of 5-azacytidine and amphotericin B, which could be involved in activating phenotype-specific genes to differentiate MSCs into myoid cells. There are good outlook on clinical treatment of illness of myatrophy using by MSCs.

OBJECTIVE To investigate the production of metallo-beta-lactamase in clinical isolates of multi-resistant Pseudomonas aeruginosa and evaluate the validity of the detection methods.METHODS The multi-resistant strains were selected by K-B method according to the standard Aloush et al recommended.The metallo-beta-lactamase phenotypes were detected by multi-disk-multi-inhibitors synergy test(MDMIST),and the genotypes of IMP and VIM gene were analyzed by PCR amplification.RESULTS A total of 192 strains of multi-resistant P.aeruginosa were selected from 1081 clinical strains.The antimicrobial agents test in these multi-resistance strains demonstrated that ciprofloxacin and piperacillin had the highest resistant rate(92.5%),and the next were aztreonam and trimethoprim-sulfamethoxazole(91.5%),the polymyxin showed sensitive in all of these strains.Sixty-seven strains of metallo-beta-lactamase phenotypes were positive and the amplification PCRs showed that 65 strains were IMP or VIM in these 192 multi-resistant strains.CONCLUSIONS The resistance mechanisms in multi-resistant P.aerugionsa present multiple and changeable.The clinical laboratory should enhance the detection of metallo-beta-lactamase in these multi-resistant strains.

Objective To investigate mupiroein resistance in Staphylococcus aureus (SAU) and the resistance to commonly used antibiotics in mupirocin-resistant strains. Methods Four hundred and ninety clinically isolated SAU strains froin January 2005 to May 2007 in the First Affiliated Hospital,Wenzhou Medical College were screened by mupirocin(5μg)disc diffusion method.Minimum inhibition concentration(MIC)and the amplification of mupA gene were performed to determine the resistance to mupirocin.Resistance to cefoxitin,gentamycin, levofloxacin, trimethoprim/sulfamethoxazole, rifampin, erythromycin, clindamycin, tetracycline and vancomycin in mupirocin-resistant strains was detected by disc diffusion method, and the amplification of mecA gene was performed to confirm the methieillin resistance among mupiroein-resistant strains.Results Twenty-seven mupirocin-resistant strains were obtained,in which 22(81.5%)were hish-level mupirocin resistant(MuH)and the rest were low-level mupirocin resistant(MuL).Among 27 mupirocin-resistant strains,24 were methicillin-resistant Staphylococcus aureus (MRSA)in which 21 were MuH and 3 were MuL strains.Drug sensitivity tests showed that the resistance to gentamycin,levofloxacin,trimethoprim/sulfamethoxazole,rifampin,erythromycin,elindamycin and tetracycline were hish among MuH and MuL strains,and most of these strains were multi-drug resistant.All strains were susceptible to vaneomycin.Conclusions Most of the clinical emerged mupirocin-resistant SAU strains are MuH and show hish resistance to commonly used antibiotics.Therefore,detection and drug sensitivity test of mupirocin-resistant strains should be strengthened in clinic practice in order to prevent it from dissemination.

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.

This paper summarizes the current research on teaching performance appraisal management of teaching hospitals at home and abroad, analyzes existing problems in teaching performance appraisal at present, and puts forward that teaching performance appraisal should be included in hospital performance appraisal. The principle of "informatization and anonymity" should be adopted to evaluate the teaching quality of clinical teachers, so that clinical teachers can participate in teaching performance appraisal management, and the indicators and weights of teaching performance appraisal can be determined scientifically and reasonably.