To investigate the contamination and drug resistance of Listeria monocytogenes in meat,158 fresh meat samples were randomly collected from meat markets in Harbin,and 23 Listeria monocytogenes strains(14.6 % positive) were isolated and characterized by Coloration medium,API reagent strip and PCR.The contamination in raw pork is the highest,with 20.00 %(14/70) detected positive.The drug resistance of isolated strains was tested against antibacterial drugs by Kirby-Barer method,and all but one(95.65 %) displayed drug resistance.The results showed the Listeria monocytogenes contamination in fresh meat was evident in Harbin and drug resistant rate was very serious.In order to reduce the drug resistance pathogens it is necessary to control and reduce the use of antibiotics in animal feedstuffs.
The csdA(cold-shock dead-box protein A) gene was amplified by PCR from the genome of Mycobacterium tuberculosis H37Rv strain,and then it was cloned into pET28a vector.The product of ligation was transformed into Escherichia coli BL21.The recombinant plasmid was confirmed by restriction digestion and DNA sequencing.The CsdA protein was expressed inducing with 0.02mmol/L IPTG.The recombinant protein was purified with Ni-resin and the immunogenicity of the recombinant protein was identified by Western-blot.The recombinant protein was confirmed to be about 64ku in size.The results showed that the recombinant expression plasmid pET28a-csdA was successfully constructed and the recombinant protein with high purity laid the foundation for further studies on the csdA protein.
This paper studies the relationship between the severe asthma as is called in Western medicine, the virtual roar asthma in Chinese medicine, and the abnormal Savda asthma in Uygur medicine. 156 patients with bronchial asthma were diagnosed and classified, according to Western medicine, Chinese medicine, and Uygur medicine, respectively. The methods used include: ① RIA analysis of interleukin-1 (IL-1), interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α), adrenocorticotropic hormone (ACTH), cortisol (CS) and growth hormone (GH) detection; ② the enzyme-linked immunoassay (ELISA) in corticotropin-releasing hormone (CRH) test. ③ the flow cytometry on lymphocyte apoptosis, CD3, CD4, CD8, CD11b, CD18 and CD62p detection. The results were treated with SPSS12.0 statistical software; measurement data were expressed by mean ± standard deviation formula; multiple group means were compared with single-factor analysis of variance, tested with α= 0.05 level. Results show that severe asthma, asthma virtual roar, abnormal Savda asthma, with respect to age, duration, saw no significant difference (P0.05), with the three symptom groups being overlapped; lymphocyte subsets and medium CD4, CD8, CD4/CD8, CD11b, CD11b/CD18 of them also saw no difference of statistical significance (P0.05); but CD62p, TNF-α and lymphocyte apoptosis of them saw some difference of statistical significance (P0.05); and CS endogenous cortisol, ACTH and CRH in severe asthma and abnormal Savda asthma were different with a statistical significance (P0.05), with no differences of statistical significance for asthma. It is concluded that severe asthma, asthma virtual roar, abnormal Savda asthma are similar to some extent, with respect to severity in patient's conditions, seniority in average age, overlapping symptoms, and immune and endocrine system disorders.
Objective To profile the types of bacteria in bile culture and study their antibiotic sensitivity pattern in patients with varying degrees of acute cholangitis and to provide data guidance for the prevention and treatment of bacterial infection. Methods The clinical data of230 patients with acute cholangitis who were admitted to our hospital from April 2010 to April 2014 were retrospectively analyzed. Classification of pathogenic bacteria and the antibiotic sensitivity test in bile cultures were performed using VITEK 2 Compact Biomerieux microbiological system. Results Within the 230 patients,172 tested positive for biliary bacteria,and the positive rate was 74. 78%(172 /230). There were 237 strains of pathogenic bacteria,including 135 strains of Gram- positive bacteria(56. 96%),comprised mainly of Enterococcus and Staphylococcus aureus,96 strains of Gram- negative bacteria(40. 51%),comprised mainly of Pseudomonas aeruginosa,Escherichia coli,and Klebsiella pneumonia,and 6 strains of fungi(2. 53%),comprised mainly of yeast. Significant difference in the positive rate of bacteria was detected between patients with mild and severe cholangitis(χ2= 4. 58,P = 0. 028). Gram- negative bacteria were the predominant isolates in patients with mild and moderate acute cholangitis,while multiple bacterial infection was more common in patients with severe cholangitis. Biliary bacteria had a higher rate of susceptibility to vancomycin,imipenem,and amikacin compared with other antibiotics.Conclusion Gram- positive bacteria are the predominant pathogens in acute cholangitis. Early detection,immediate intervention,and accurate drug susceptibility test should be implemented at regular intervals to direct effective clinical therapy.
To investigate the biofilm formation of the avian pathogenic Escherichia coli by establishing a in vitro qualitative model and quantitative adhesion test. A wild type pathogenic E. coli was cultivated in flat-bottom 96-well polystyrene microplate substratum with 5% TSB medium under different cultivation time,mediums,substratums and nutrients. Biofilm was well developed at 37 ℃ after 48 hours cultivation. The formation period included an initial adhesion to the surface at 8 hours,the development of micro-colonies at 24 hours,the conglutination of micro-colonies at 36 hours,the mature of biofilm at 48 hours,and the desquamation from the substratum,and followed by a new formation circulation. It was also showed that sugars,sodium chloride and Mg2+ would enhance the development of the biofilm and the adherent ability of the bacteria. The results may provide a basis for clinical therapy to the bacterial biofilm disease.
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