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目的:为了解黄连在动物体内有无消除R质粒的作用,探讨临床应用的可能性,本课题以多重耐药性的大肠埃希菌E.102株为靶细菌,建立一种以去污染的小白鼠为模型的R质粒体内消除实验方法。方法:首先用四环素,红霉素和利福平对实验用小鼠进行肠道细菌脱污染,建立去污染小白鼠动物模型,然后将靶细菌E.102株菌液经口感染,并随机分组,试验组小鼠每日经灌胃法给黄连1.2mg,对照组不给黄连。在给药后24、48、72h将小鼠处死,解剖取其盲肠内容物,以SS平板划线培养24h,随机挑取单个菌落用影印培养法接种于选择平板上(分别含有不同的抗生素,浓度分别为四环素20mg/L,链霉素40mg/L,氨苄青霉素20mg/L),检查细菌对药物的敏感性。凡是在营养琼脂平板上生长,而在含有抗生素的选择平板上不生长的菌落,即为消除了R质粒的菌落。结果:去污染小鼠动物模型的建立:在口饲四环素、红霉素和利福平3d后,解剖小鼠,取盲肠内容物在SS平板上划线培养48h,未见细菌生长,证明小鼠肠道脱污染成功。经口服大肠埃希菌液3d后,解剖小鼠盲肠内容物,用SS平板分离培养,均得到纯的大肠埃希菌培养物,证明靶细菌定殖成功。黄连在去污染小鼠体内对R质粒的消除?
Objective: To understand whether Rhizoma Coptidis can eliminate R plasmid in animals and to explore the possibility of clinical application, this study aims to use multi-drug resistant E. coli. 102 strains were used as target bacteria to establish an experimental method for in vivo elimination of R-plasmids using decontaminated mice as a model. METHODS: First, tetracycline, erythromycin, and rifampicin were used to decontaminate the experimental mice and the decontaminated mouse model was established. The target bacteria were then used. 102 strains were orally infected and randomly divided into groups. The mice in the experimental group were given 1.2 mg of Coptis Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma Rhizoma perfusion by daily gavage. The mice were sacrificed at 24, 48, and 72h after administration. The contents of the cecum were dissected and cultured on SS plates for 24 hours. A single colony was randomly picked and inoculated on the selection plate by photopheresis culture method (containing different antibiotics, respectively). The concentrations were tetracycline 20 mg/L, streptomycin 40 mg/L, ampicillin 20 mg/L), and the sensitivity of the bacteria to the drug was examined. Any colony that grows on a nutrient agar plate and does not grow on a selection plate containing antibiotics is a colony that eliminates the R plasmid. RESULTS: Establishment of a decontaminated mouse animal model: After oral administration of tetracycline, erythromycin and rifampicin for 3 days, the mice were dissected and the cecal contents were streaked and cultured on SS plates for 48 h. No bacterial growth was observed, demonstrating that the mice were small. Intestinal decontamination was successful. After oral administration of Escherichia coli liquid for 3 days, the contents of the caecum of the mice were dissected and separated and cultured using SS plates. Pure E. coli cultures were obtained, which proved that the target bacteria were successfully colonized. Coptis eliminates R plasmid in decontaminated mice?