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Objective To investigate the drug resistance of Staphylococcus haemolyticus and provide basis for selection of clini‐cal drugs .Methods A total of 59 strains of Staphylococcus haemolyticus were isolated from patients ,and analyzed the clinical dis‐tribution and drug resistance of them .Results Staphylococcus haemolyticus were mainly isolated from the sputum and oropharyn‐geal swabs in pediatric and ICU .The patients were mainly newborns .Staphylococcus haemolyticus mainly included methicillin re‐sistant Staphylococcus haemolyticus (MRSH) ,accounting for 89 .8% (53 strains) .MRSH showed high‐drug resistance and multi‐drug resistance .However ,it was sensitive to vancomycin ,teicoplanin and linezolid ,the antibiotic sensitive rate was 100 .0% .Conclu‐sion Staphylococcus haemolyticus is an important coagulase negative staphylococci ,mainly including MRSH ,which shows high‐drug resistance and multi‐drug resistance .
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Objective To develop a quantitative immunohistochemistry assay for duck hepatitis B virus core antigen (DHB-cAg)in duck liver tissue.Methods By comparison with no repair antigen and repair antigen with high pressure,microwave and trypsin,the best solution of antigen retrieval was determined.By optimizing the parameter of image acquisition and de-ducting blank area,mean density of yellow areas was calculated using Image-Pro Plus 6.0 software.Using the assay devel-oped to determine the level of DHBcAg in liver tissue from duck infected by DHBV,anti-DHBV activity of DHBcMAb-TAT PTD conj ugate was examined.Results SABC method with no repair antigen was selected,which was better than other methods.DHBcAg expression in duck liver tissue could be objectively and accurately quantified by setting Image-Pro Plus 6.0 software parameters and calculating mean density of yellow areas.By comparison with the differences between mean densityat baseline of treatment and end of treatment,it was showed that DHBcMAb-TATPTD conjugate treatment dose-de-pendently reduced the levels of DHBcAg in liver tissue,which show that the assay developed could effectively evaluate the anti-DHBV activity of agent.Conclusion The immunohistochemistry assay developed in this study can objectively and accu-rately evaluate the level of DHBcAg in duck liver tissue.
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Objective Try to know the prevalence rate of ESBL-s produced by Eseherichia coil in our hospital from 2003 to 2007.Methods Antibiotic susceptibility were determined by K-B disc diffusion.Double-disk synergy tests performed on Mueller-Hinton agar plates was used for ESBL confirmation.Results There was 23.8% of our isolates can produce ESBLs in 2003,25.4% in 2004,31.3% in 2005,36.6% in 2006 and 44.9% in 2007,respectively.The situation of antibiotic resistance is more severe.Conclusion The antibiotic resistance should be well surveyed and the ESBLs must be routinely assayed.
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Objective To assess the differences in the specificity and the sensitivity in detecting specific IgM by indirect ELISA and capturing-ELISA methods,and to analyze the possible causes of those differences.Methods The HBc-IgM and EHF-IgM level in serum samples from activity Hepatitis B patients and hemorrhagic fever patients with renal syndrome were determined by both indirect-ELISA and capturing-ELISA methods,and the differences in the positive rate,positive threshold value and the specificity between those two methods were compared. Results Specific murine IgM diluted in PBS were detected by indirect-ELISA and capturing-ELISA,and the specificity and sensitivity of those two methods were similar.Results showed that sensitivity of indirect-ELISA was lower than capturing-ELISA in detecting specific IgM in serum from patients with activity Hepatitis B and hemorrhagic fever patients with renal syndrome.The IgM level in hepatitis B and hemorrhagic group treated with thioglycol became negative detected by those two methods,suggesting that both of them have the anti-IgM epitope-specificity.Cross reaction results demonstrated that the reagent detecting IgM from the two methods had the specificity to the specific antigen. Conclusion It is recommended to detect IgM level by capturing-ELISA method for the early diagnosis of acute infectious disease,pathological changes,immune reaction and prognoses of chronic continuous infectious disease.Applying the indirect-ELISA method to detect IgM level to diagnose acute infectious disease is to discuss in the future.
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Objective To establish better reaction factors for rep-PCR and to investigate if the extended spectrum ?-lactamase(ESBL)+ Escherichia coli isolated from different patients in the respiratory ward have the same origin by using rep-PCR.Methods The ESBL confirmation was taken by double disc confirmatory test.The susceptibility testing was performed with K-B test.By applying the widespread enterobacter repetitive intergenic consensus as a primer,the stains were typed by rep-PCR following electrophoresis in agarose gel.Results The analysis of the PCR productions indicated that it would create useful DNA ban-prints and all these 22 ESBL(+)Escherichia coli strains were of three origins.Conclusion The method of rep-PCR is practical for epidemiological research in nosocomial infection.
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Objective To construct a recombinant lentiviral vector for HCV core-Ant and then study its effect on the transdifferentiation of hepatic stem cells. Methods The HCV core-Ant was obtained by PCR of two primers template with each other and T-vector cloning method,and then subcloned to pLenti6/V5-D-TOPO. The restriction endonuclease and T4 DNA ligase were used to construct the vector. pLenti6/V5-D-HCV core-Ant and the ViraPowerTM PackagingMix (containing three packaging plasmids pLP1,pLP2 and pLP/VSVG) were cotransfected into 293ET cells to produce replication in competent lentivirus after transfection. The viral supernatant on 293T cells was collected. The expression of the lentiviral vector containing HCV core-Ant in Hela cells was measured by immunohistochemistry. Then we constructed the lentiviral vector containing green flurosecent protein by the similar method,and the titers were determined. Results HCV core-Ant was identified and analyzed by restriction enzyme digestion and sequencing,respectively. The expression of the recombinant lentiviral vector plasmid containing HCV core-Ant in Hela cells was confirmed by immunohistochemistry. The 3T3 cells transfected the lentiviral vector containing green flurosecent protein were found to show strong expression of GFP,which confirmed that the four-plasmid system of the lentiviral vector was successfully constructed. Conclusion The recombinant lentiviral vector expressing HCV core-Ant was successfully constructed by molecular cloning and recombination techniques in vitro,which will be beneficial to guiding further study on gene therapy of cancer.
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Objective To investigate the clinical value of fecal calprotectin as a marker for diagnosis of ulcerative colitis(UC).Methods ELISA assay was used to measure the concentrations of calprotectin in feces of 30 UC patients and 20 healthy controls.The disease activity of UC was determined by Truelove Witts histological criteria.Results The level of calprotectin in feces was significantly higher in feces with active UC(578?g/g) than with inactive UC(8.4?g/g,P0.05),but significant difference existed between each active grade and inactive phase of UC.The results revealed siginificant correlation between the endoscopic grades of disease activity and fecal calprotectin concentration(r=0.85,P