By using methods of forecast research,system science,comparison research and abstract research,main deficiencies and practical problems in requisitioning civilian medical equipment are analyzed.It is suggested that we should regard the health service's demand as direction,the preparation as premise,the information construction as fundamentality,the lawmaking as guarantee,to improve and perfect the work about the requisition of civilian medical equipment and improve the quality and benefit of mobilization.

The dependability of medical equipment directly affects the quality and benefit of health service. The medical equipment dependability of our army apparently lags behind foreign armies. It is urgent for us to improve the dependability of the medical equipment.Our army must innovate the research and production as well as purchase of the medical equipment; enhance the inherent dependability and the use dependability of the medical equipment, actualize research on non-effectiveness of the medical equipment, improve the production design and enhance the skill of the operators.

Objective To clone, express, purify and evaluate the immunoreactivity of the recombinant protein Tp0844 of Treponema pallidum (Tp), and to screen major Tp proteins with high host reactivity. Methods The Tp0844 gene sequence was obtained through bioinformatics analysis. A prokaryotic expression vector of the Tp0844 gene was constructed and transformed into E. coli BL21 followed by isopropyl-1-thio-β-D-galactopyranoside (IPTG)induction for the expression of the recombinant protein Tp0844. Nickel-NTA affinity chromatography columns were utilized to purify the recombinant protein, and Western blotting was performed to evaluate the reactivity of the recombinant protein with sera positive or negative for anti-Tp IgG antibodies. Results The recombinant prokaryotic expression vector PET-30a (+)-Tp0844 was successfully constructed. After IPTG induction, a soluble recombinant protein with a relative molecular mass of about 43 000 was highly expressed, and purified by affinity chromatography. Western blotting showed that the Tp0844 recombinant protein specifically reacted with anti-Tp IgG antibody-positive sera, but not with anti-Tp IgG antibody-negative sera. Conclusions The soluble recombinant protein Tp0844 has good immunoreactivity, and can serve as a candidate antigen for investigation into the pathogenesis of syphilis.

OBJECTIVE To develop the strong and the specific multi-epitope antigen for the exploiting diagnosis of Treponema pallidum.METHODS The immuno-dominant epitopes of Tp0453 and Tp17 were amplified by PCR respectively,and subcloned into the expression vector pQE32 to generate multi-epitopes recombinant plasmid pQE32/Tp0453-17.The recombinant protein was expressed in Escherichia coli M15.The immunoresponse of recombinant fusion protein was analyzed by Western blot.RESULTS The multi-epitopes recombinant plasmid was successfully constructed,enzyme digestion analysis and sequencing showed that the inserted target genes were Tp0453 and Tp17 gene,compared with the gene reported by GenBank,it had 100% similarity;SDS-PAGE analysis showed the recombinant plasmid could be expressed in M15,its relatively molecular mass(Mr) of expressed product was about 52.0?103.The Western blot result showed the recombinant protein could be recognized by anti-T.pallidum positive serum.CONCLUSIONS The expressed multi-epitopes recombinant antigen showed excellent immunoresponse.The results lay the foundation for research on development of quick diagnostic kit applying to detection of T.pallidum infection.

OBJECTIVE To investigate the distribution of clinical bacterial isolates and the change in antibiotic resistance spectrum in our hospital from 2005 to 2007.METHODS Data of bacterial susceptibility testing of clinical isolates from the Second Affiliated Hospital in of University of South China from 2005 to 2007 were collected and analyzed by software WHONET25.Results were assessed according to the National Committee for Clinical Laboratory Standards(NCCLS) of America issued in 2005.RESULTS The amount of Gram-negative bacteria decreased and of Gram-positive bacteria increased during this period.The proportion of coagulase negative Staphylococcus(CNS) had been increasing and reached 21.7% in 2007.The proportions of Staphylococcus aureus decreased from 17.6% in 2005 to 13.0% in 2007.Escherichia were the top two bacteria in 2007.The drug resistance rate of staphylococci against penicillin and erythromycin was more than 92.2% and 52.2%,respectively.The oxacillin resistance rate of CNS was 74.5%,significantly higher than that of S.aureus(16.5%).Drug resistance rate of Enterococcus to vancomycin was 1.1%.Gram-negative bacteria were found resistant to meropenem and imipenem.The resistance rate to ampicillin of Klebsiella and Escherichia was very high.CONCLUSIONS The variation of drug resistance and distribution of clinical bacterial isolates in our hospital are related to the improper use of antibiotics.It is very important to select antibiotics correctly according to the results of antibiotics susceptibility tests.

The antibacterial peptide CM4 having potent antifungal activity on inhibitiong the cell wall regeneration of Saccharomyces cerevisiae protoplasts.When the peptide increased,the ratio of the regenerated colonies drop obviously.To study the antifungal mechanism of the antibacterial peptide,fluorescence\|labeled peptide mixted with the protoplast of yeast,then confocal laser scanning microscopy were performed.The results indicated that the peptides interactted with the protoplast membrane and damaged the structure of the membrane,then the permeation of protoplast changed.Finally the protoplasts with the peptide failed to regenerate the cell walls leading to killing the cell.

Objective To investigate the prevalence of A2058G or A2059G mutation within 23S rRNA in Treponema pallidum (Tp) from primary syphilis patients chancre samples. Methods Simple PCR was used to screen the positive samples containing Tp DNA. Nested PCR was adopted to amplify the region of the Tp 23S rRNA and the purified amplicons were digested by restriction endonuclease MboⅡand Bsa I respectively and sequenced. Results 39 qualified samples were obtained from 43 chancre samples and all of them were found harboring the A2058G mutation, whereas the A2059G was not detected. Conclusion High frequency of the A2058G mutation within 23S rRNA implicated in macrolide resistance emerges in the circulating Tp in Hengyang. Therefore, macro-lide antibiotics such as azithromycin should be cautiously used as an optional therapy for syphilis.

Sepharose 6B was activated by epichlorohydrin in the strong base condition, and then reacted with solution of iminodiacetic sodium. The arms of IDA were conjuncted to the activated Sepharose 6B. Then the products were reacted with the solution of NiSO4. The arms of IDA were chelated with Ni2+,and the chelating resin―Ni2+-IDA could be prepared. The physicochemical indexes and performance in purifying protein of the expressing product were assayed with atomic absorption method and purifying aimed protein-human B lymphocyte stimulator(hBLyS) from the expressing products in E.coli. The results indicated that the performance of made gel is very good, and its price is less than 1/10 of that of commodity gel.

Objective To evaluate the value of a Treponema pallidum (TP) recombinant protein TP0993 in the serodiagnosis of syphilis.Methods A bioinformatics method was used to obtain the sequence of TP0993 gene.The open reading frame (ORF) without upstream non-coding region of TP0993 gene was ligated into the expression vector PET-28a (+),which was then transformed into Escherichia coli Rosetta.Isopropyl-β-d-thiogalactoside (IPTG) was used to induce the expression of TP0993 protein.The expressed protein was purified with nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography.Western blot was performed to evaluate the immunoantigenicity of the protein.New Zealand rabbits were immunized with the recombinant protein for immunogenicity evaluation.Indirect enzyme linked immunosorbent assay (ELISA) was developed by using the purified recombinant protein to coat microwell plates.Anti-TP antibodies were detected by the established ELISA and TP particle agglutination assay (TPPA) in 480 clinical serum samples.Results The prokaryotic expression vector PET-28a (+)-0993 was successfully built,and a fusion protein with a relative molecular weight of about 34 000 Da was attained after IPTG-induced expression and purification.Western blot proved that the recombinant protein could specifically react with clinical sera positive for anti-TP IgG antibodies.Specific humoral response was elicited in New Zealand rabbits by the recombinant protein.Compared with TPPA,the established indirect ELISA showed a sensitivity of 88.3％ and a specificity of 85.8％.There was a consistency of 86.5％ between the indirect ELISA and TPPA.Conclusion The expressed recombinant protein showed favorable immunocompetence,and may serve as a candidate antigen for serodiagnosis of syphilis.

Objective To clone.and express Tp0453 outer membrane protein of Treponema pallidum and develop an indirect ELISA for sero diagnosis of syphilis. Methods The immuno-dominant epitope of Tp0453 was amplified by PCR and subcloned into the expression vector pQE32.The recombinant protein was expressed in E.coli M15 and purified with Ni-NTA affinity chromatography columns. Indirect ELISA was developed to detect the antibody to Tp in human sera.Results 60 control sera was tested by ELISA.The sensitivities was 100%(30/30), and the specificities was 100%. While detecting uninfected and infected T. pallidum human sera, the sensitivities of ELISA was 96.8% compared with the results of the TPPA tests, and the specificities was 100% when the results of ELISA was compared with those of the TPPA test. The concordance of results between the ELISA test and the TPPA test was 98.2%.Conclusion The recombinant Tp0453 outer membrane protein showed excellent immuno-reactive activity, and were suitable for development of ELISA for sero-diagnosis of syphilis.