中国人肠道产甲酸草酸杆菌甲酰辅酶A转移酶基因的分离、克隆和鉴定

目的 克隆产甲酸草酸杆菌甲酰辅酶A转移酶基因（frc），检测其在真核细胞中的表达。方法 用聚合酶链式反应技术从产甲酸草酸杆菌基因组DNA中扩增frc基因片段并插入真核表达载体获取重组质粒pEGFP-frc，通过限制性内切酶酶切电泳和测序鉴定重组质粒。重组质粒转染人胚。肾293细胞，检测frc基因在mRNA和蛋白水平上的表达。被转染细胞在含草酸培养液中培养，检测0～72h培养液中草酸浓度的变化。结果 frc基因的真核表达载体构建成功。转染293细胞后24～72h，可观察到明亮的绿色荧光，在mRNA和蛋白水平上可检测到frc基因在真核细胞中的表达。转染pEGFP-frc细胞12～72h培养液中草酸浓度明显低于转染空载体的细胞（P〈0．05）。结论 中国人肠道产甲酸草酸杆菌中可分离出frc基因；frc基因可在真核细胞293细胞中表达并使293细胞获得代谢草酸的能力。     

泌尿系结石的基因诊断和治疗

泌尿系结石是泌尿系统最常见的疾病之一，在我国一般人群中发病率高达1％-10％。其中约25％的患者需住院治疗，在泌尿外科住院患者中占居首位。近年来，我国泌尿系结石的发病率有增加趋势，是世界上三大结石高发区之一。     

Cloning and Identification of frc Gene from Oxalobacter Frmigenes

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chi- nese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.     

泌尿男生殖系肿瘤第七版TNM分期的变化及解读

目前,恶性肿瘤已成为威胁人类健康与生命的主要疾病之一.在发达国家,恶性肿瘤是国民死亡的首要原因.在我国,恶性肿瘤占城市居民死因的第一位,在农村居民的死因中也呈上升趋势.根据世界卫生组织国际癌症研究署(International Agency for Research on Cancer,IARC) 2011年的统计资料,全球每年新发恶性肿瘤1270万例,死于恶性肿瘤者760万例[1].恶性肿瘤在其病程发展的不同阶段,预后相差甚远,早期常常可以取得满意的治疗效果,甚至完全治愈,而晚期肿瘤目前缺乏有效的治疗手段,很多患者只能接受姑息性治疗.因此,一个准确的分期系统对选择治疗方式和判断预后有指导性意义.