目的 分析2011—2016年间铜绿假单胞菌分离株的耐药性及变迁情况, 为临床合理用药提供科学依据。方法 对2011年1月—2016年12月广州市第一人民院患者各类标本中分离到的铜绿假单胞菌2 257株进行细菌鉴定及药敏试验,并对耐药性变迁进行统计分析。结果 铜绿假单胞菌在痰液标本中的检出率最高为56.9％;6年铜绿假单胞菌平均耐药率以妥布霉素最低,为9.9%,对哌拉西林/他唑巴坦、头孢吡肟、头孢他啶、左氧氟沙星、环丙沙星、亚胺培南、庆大霉素等药物的耐药率均＜20%,在2013年耐药率最低,此后三年逐年上升。结论 铜绿假单胞菌对广州市第一人民院常用抗生素的耐药率在近3年呈逐年上升趋势, 临床医师应根据药敏结果合理选择抗菌药物, 以提高疗效和减缓耐药菌的产生。

Objective To analyze the changes of drug resistance of Pseudomonas aeruginosa (Pae) and to provide basis for the use of antibiotics in clinic. Methods 2 257 strains of Pae were cultured and isolated in the First People Hospitalof Guangzhou from 2011 to 2016, API bacterial identification system was applied to carry out bacterial identification and K-B method was used for drug sensitivity analysis. Results Most of the Pae (56.9％) were detected from the sputum specimen. It showed the highest sensitivity to tobramycin. The drug resistance of Pae to piperacillin/tazobactam, cefepime, ceftazidime, levofloxacin, ciprofloxacin, imipenem and gentamicin in 2013 was the lowest and has been increasing year by year. Conclusion Pseudomonas aeruginosa isolated in our hospital showed a rising trend of clinical drug resistance in the past three years. It was of the top priority for clinicians to use antibiotics rationally to retard the production of drug resistant strains.

目的 构建重组pEGFP-C3-HCVc真核表达载体,并建立稳定表达HCVc基因的肝内胆管癌细胞株RBE-core。方法 采用PCR钓取目的基因HCVc,并克隆入pEGFP-C3的多克隆位点,构建pEGFP-C3-HCVc重组质粒。经过双酶切及测序验证后,采用脂质体将pEGFP-C3-HCVc质粒转染到RBE细胞中,经2周G418 (200 μg/mL) 筛选后进行单克隆挑选及扩大培养,建立稳定表达HCVc的胆管癌细胞株RBE-core。采用RT-PCR和Western blot验证HCVc在RBE-core中的表达情况。结果 PCR成功钓取HCVc基因,大小约573 bp,并插入pEGFP-C3载体HindⅢ和BamHⅠ多克隆位点;双酶切及测序证实目的基因HCVc正确连接到pEGFP-C3的多克隆位点。RT-PCR和Western blot分别在573 bp处和34 KD左右检测到相应的阳性条带。结论 成功构建重组质粒pEGFP-C3-HCVc,并在胆管癌细胞RBE中获得稳定表达。

Objective To construct a recombinant plasmid of pEGFP-C3-HCVc containing hepatitis C virus core protein, and establish the HCVc-expressing cell line RBE-core. Methods The HCVc gene was amplified by PCR and cloned into HindⅢ and BamHⅠsite of pEGFP-C3 plasmid. The recombinant plasmid of pEGFP-C3-HCVc was confirmed by sequencing. RBE cells were transfected with the recombinant plasmid by using Lipofectamine 2000, and then performed G418 (200 μg/mL) selection after 2 weeks. The expressing of HCVc gene in RBE cells was confirmed by RT-RCR and western blot. Results The recombinant plasmid of pEGFP-C3-HCVc was successfully constructed. RT-PCR and western blot detected a 573bp and 34KD bland, indicating the stably expressing of HCVc in RBE cells. Conclusion The recombinant plasmid of pEGFP-C3-HCVc is stabled expressing in RBE cells,which provides support for the further study.