毛白杨4CL-like基因的克隆与原核表达分析

为得到毛白杨4CL-like基因并对其原核表达特性进行分析,利用毛白杨叶片的总RNA反转录得到的总cDNA为模板,利用PCR技术克隆得到毛白杨4CL-like基因(登录号:XP_002315339.1)。利用MEGA软件对4CL-like基因及其他9个来自不同物种的4CL基因进行进化树分析。最后通过构建pET30-4CL-like原核表达载体对该基因进行原核表达分析。结果显示,克隆得到的4CL-like基因CDS区全长为1 758 bp,编码585个氨基酸。系统进化分析表明,该基因与葡萄中的4CL-like5有很高的同源性。另外,SDS-PAGE分析表明,4CL-like基因在大肠杆菌BL21中成功表达,所表达融合蛋白的分子量约为60 ku。

Effect of ezetimibe on the prevalence of cholelithiasis

AIM： To investigate the prevalence of cholelithiasis among patients treated with ezetimibe. METHODS： A retrospective, case-control study based on computerized medical records from patients of the Clalit Health Services, Sharon-Shomron region, from 2000 to 2009. Patients 20-85 years of age, who had been treated with ezetimibe and statins or statins only for at least 6 too, and who had an abdominal ultrasound were included in the study. Collected data included age, gender, ezetimibe treatment duration, presence of hypothyroidism or diabetes, and existence of cholelithiasis as determined by ultrasound. Ex- cluded were subjects after gallbladder resection, with hemolysis, myeloproliferative or inflammatory bowel diseases, and those treated with ursodeoxycholic acid and fibrates. Patients treated with statins and ezeti- mibe （study group） were compared to patients treated with statins only （control group）. RESULTS： The study group included 25 patients and the control group 168. All patients in the study were treated with statins. The study group included 13 males （52%） and 12 females （48%）, the control group 76 males （45%） and 92 （55%） females （P = 0.544）. The groups did not differ in age （mean age： 68 ± 8 years, range 53-85 years vs mean age： 71±8 years, range 51-85 years; P = 0.153） or in the rate of dia- betic and hypothyroid patients [11 （44%） vs 57 （33%）, P = 0.347 in the study group and 5 （20%） vs 23 （14%）, P = 0.449 in the control group, respectively]. Patients in the study group were treated with ezetimibe for an average of 798±379 d. Cholelithiasis was found in 4 （16%） patients in the study group and in 33 （20%） patients in the control group （P = 0.666）. CONCLUSION： Ezetimibe does not appear to influ- ence the prevalence of gallstones.

姜黄素联合胡椒碱在预防小鼠胆囊胆固醇性结石中的作用

目的探讨姜黄素联合胡椒碱在预防高脂饮食小鼠胆囊胆固醇结石形成中的作用。方法 50只C57BL6小鼠全部喂给高脂饮食,同时每天灌喂500或1 000 mg/kg姜黄素,每天加或不加20 mg/kg胡椒碱,共4周。统计胆囊结石成石率,收集血液、胆汁、胆囊、肝脏以及小肠,测量胆囊容积,称量肝脏质量,检测血液及胆汁生化指标,对小鼠肝脏及胆囊进行HE染色,同时对胆囊行天狼猩红染色,real-time PCR、Western blot方法检测小鼠小肠Neimann-Pick C1-like 1(NPC1L1)及固醇调控元件结合蛋白-2(SREBP2)表达。结果与单纯高脂饮食喂给相比,姜黄素联合应用胡椒碱可降低70%高脂饮食所致的小鼠胆囊胆固醇性结石的形成,同时降低胆固醇〔(2.89±0.14)mmol/L比(6.86±0.16)mmol/L,P<0.05〕及甘油三酯〔(0.84±0.09)mmol/L比(1.59±0.10)mmol/L,P<0.05〕,还可降低小鼠小肠组织中NPC1L1 m RNA(0.46±0.023比1.00±0.014,P<0.05)及蛋白表达(0.47±0.01比1.00,P<0.05),同样降低小鼠小肠组织中SREBP2 m RNA(0.80±0.028比1.00±0.061,P<0.05)及蛋白表达(0.77±0.05比1.00,P<0.05)。结论姜黄素能够预防高脂饮食小鼠胆囊胆固醇性结石的形成,NPC1L1及SREBP2可能参与了这个过程,胡椒碱可以增加姜黄素的生物利用度,增强其作用。