miR-513a-5p慢病毒载体的构建及其对人骨肉瘤细胞放疗敏感性的影响

目的:构建miR-513a-5p慢病毒过表达载体，转染人骨肉瘤细胞株，观察miR-513a-5p对人骨肉瘤细胞放疗敏感性的影响。方法:PCR法扩增人miR-513a-5p基因，克隆入pLentis-CMV-GFP-MCS-PGK-PURO载体获得重组质粒pLentis-miR513a，双酶切鉴定并测序后将正确的重组质粒和对照质粒转染293FT细胞制备慢病毒，分别转染骨肉瘤HOS和U2OS细胞，qRT-PCR法及荧光显微镜鉴定转染结果。克隆形成实验、MTT法检测miR-513a-5p高表达HOS和U2OS细胞在X射线照射下细胞存活情况。结果:双酶切及测序结果确定成功构建miR-513a-5p慢病毒载体pLentis-miR513a。qRT-PCR结果提示，转染骨肉瘤细胞株后miR-513a-5p表达显著升高。克隆形成实验结果显示miR-513a-5p高表达后骨肉瘤细胞在X射线照射下细胞增殖减慢。MTT结果提示miR-513a-5p高表达骨肉瘤细胞经X射线照射后细胞存活减少。结论:成功构建了miR-513a-5p慢病毒载体，建立了高效稳定表达miR-513a-5p的骨肉瘤细胞株，高表达miR-513a-5p能显著增加X射线照射后骨肉瘤细胞的放疗敏感性。     

护理教学查房在ICU教学中的临床应用效果分析

目的探讨护理教学查房在ICU教学中的临床应用效果。方法选定2018年1月-2019年1月来本院实习的100例学生,按照教学查房方法分为对照组(50例)与观察组(50例),前者采用传统教学查房方法,后者采用针对性教学查房方法。通过出科操作考试及理论考试后,比较2组实习学生的操作考核成绩、理论考核成绩指标。结果教学结束,观察组实习学生的操作考核成绩(94.11±3.54)分、理论考核成绩(85.74±5.39)分均高于对照组实习学生(P<0.05)。结论针对性教学查房方法可有效提高实习学生的操作考核成绩、理论考核成绩,改善其教学质量。     

人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达

目的：克隆人肠道病毒71型（EV71）及柯萨奇病毒A组16型（CA16）的VP1蛋白编码基因，构建相应真核表达质粒在体外进行重组表达，检测其细胞内定位，为EV71及CA16的疫苗研究提供基础。方法：通过PCR扩增分别获取EV71及CA16的VP1编码序列，插入pcFlag载体，分别构建EV71及CA16 VP1与FLAG标签融合的真核表达质粒；瞬时转染人胚肾293T细胞及横纹肌肉瘤RD细胞，Western blot法检测融合蛋白的表达，免疫荧光检测融合蛋白的细胞内定位情况。结果：测序表明两种来源VP1蛋白的重组表达质粒构建正确；分别转染两种不同的细胞后，均可通过Western blot检测到相应蛋白表达；免疫荧光检测显示两种毒株的VP1均表达在细胞质中。结论：外源融合表达EV71或CA16 VP1蛋白的真核表达质粒构建成功；所构建质粒分别转染细胞后，均可在细胞质内检测到VP1融合蛋白的表达。     

可切除肺癌预后预测模型的构建及生存分析

目的 探讨构建的可切除肺癌预后预测模型在患者生存及预后预测中的价值。方法 选择山西省肿瘤医院2007年1月至2018年9月原发性肺癌患者2 267例,患者均行一次肺癌手术治疗,无第二原发肿瘤。选取性别、年龄、职业、肿瘤部位、病理类型、手术路径、手术方式、肿瘤分期、治疗方案为预后影响因素。采用Cox比例风险模型构建预后指数(PI)方程,计算每例患者的PI值。根据PI值的不同范围,划分低、中、高危预后组,对各组生存情况进行评估。结果 性别(RR=0.684,P=0.001)、年龄(RR=0.591,P<0.01)、职业(RR=1.439,P=0.001)、病理类型(RR=3.694,P<0.01)、手术路径(RR=0.734,P=0.001)、肿瘤分期(RR=0.352,P=0.007)为可切除肺癌患者预后独立影响因素。其中,女性、≤65岁、胸腔镜手术、肿瘤分期Ⅰ期为预后保护因素,其预后不良风险分别降低31.6%、40.9%、26.6%、64.8%;农民、腺鳞癌为预后危险因素,其预后不良风险分别增加43.9%、269.4%。PI方程为:∑βixi=-0.380 X1-0.526 X2+0.364 X31+1.307 X55-0.309 X6-1.045 X81(X1代表性别,X2代表年龄,X31代表职业为农民,X55代表病理类型为腺鳞癌,X6代表手术路径,X81代表肿瘤分期Ⅰ期)。PI<-1为低危组,PI≥-1且≤-0.5为中危组,PI>-0.5为高危组。1、3、5年生存率低危组分别为96.8%、87.0%、77.9%,中危组分别为91.8%、82.2%、61.7%,高危组分别为86.5%、61.7%、50.3%,各组间生存率差异具有统计学意义(P<0.05)。结论 可切除肺癌预后预测模型能够预测可切除肺癌患者的预后风险及相应生存率,帮助临床医师评估预后及制订后续治疗方案。     

Recent advances in preclinical in vitro approaches towards quantitative prediction of hepatic clearance and drug-drug interactions involving organic anion transporting polypeptide (OATP) 1B transporters

Hepatic uptake mediated by organic anion transporting polypeptide (OATP) 1B1 and 1B3 can serve as a major elimination pathway for various anionic drugs and as a site of drug-drug interactions (DDIs). This article provides an overview of the in vitro approaches used to predict human hepatic clearance (CL_h) and the risk of DDIs involving OATP1Bs. On the basis of the so-called extended clearance concept, in vitro–in vivo extrapolation methods using human hepatocytes as in vitro systems have been used to predict the CL_h involving OATP1B-mediated hepatic uptake. CL_h can be quantitatively predicted using human donor lots possessing adequate OATP1B activities. The contribution of OATP1Bs to hepatic uptake can be estimated by the relative activity factor, the relative expression factor, or selective inhibitor approaches, which offer generally consistent outcomes. In OATP1B1 inhibition assays, substantial substrate dependency was observed. The time-dependent inhibition of OATP1B1 was also noted and may be a mechanism underlying the in vitro–in vivo differences in the inhibition constant of cyclosporine A. Although it is still challenging to quantitatively predict CL_h and DDIs involving OATP1Bs from only preclinical data, understanding the utility and limitation of the current in vitro methods will pave the way for better prediction.     

Lower body mass index and higher height are correlated with increased varicocele risk

To evaluate the anthropometric indexes in subjects with varicocele compared to controls and the incidence of varicocele in different body mass index (BMI) groups for the purpose of exploring the association between varicocele and anthropometric indexes. A comprehensive literature search was conducted by using PubMed, MEDLINE, EMBASE databases and Cochrane Library up to February 2019. A systematic review and meta‐analysis was conducted by STATA, and Newcastle–Ottawa Scale was utilised for assessing risk of bias. Ultimately, 13 articles containing seven case–control studies and six cross‐sectional studies with 1,385,630 subjects were involved in our study. Pooled results demonstrated that varicocele patients had a lower BMI (WMD = ?0.77, 95% CI = ?1.03 to ?0.51) and a higher height than nonvaricocele participants, especially in grade 3 varicocele patients. Subgroup analyses showed that normal BMI individuals had a higher risk of varicocele than obese or overweight individuals and a lower risk than underweight individuals. In conclusion, this study indicates that varicocele patients have a lower BMI and a higher height than nonvaricocele participants. Moreover, men with excess bodyweight have a lower incidence of varicocele compared to normal weight or underweight people. That is to say, high BMI and adiposity protect against varicocele and high BMI is associated with a decreased risk of varicocele.     

低密度脂蛋白在2型糖尿病骨质疏松中的作用研究进展

2型糖尿病及骨质疏松已成为我国最主要的慢性代谢性疾病。2型糖尿病常伴有血脂紊乱，常表现为低密度脂蛋白升高，而越来越多的研究表明血脂通过不同的方式影响骨代谢，其机制可能是通过抑制骨髓间充质干细胞成骨分化，通过RANK/RNAKL/OPG信号通路及炎症反应调节破骨细胞等方式调节骨代谢。     

抑郁症大鼠模型海马区及前额叶组织BDNF因子、Trkb蛋白表达的实验研究

目的:观察抑郁症大鼠模型脑组织不同部位神经营养因子水平的变化及抗抑郁药物使用后改变。方法:将成年SD大鼠30只分为正常对照组、模型对照组、药物干预组,每组10只,雌雄对半;正常组正常养殖,模型组采用慢性不可预知应激结合孤养方式制备抑郁模型大鼠4周。药物干预组采用模型组造模过程4周后给予药物(氟西汀)给入,自实验开始之后每周观察大鼠体重、糖水消耗、旷场实验指标的变化,最后采用荧光定量PCR法观察大鼠脑部BDNF因子及Trkb的表达。结果:与正常对照组大鼠相比,模型组大鼠体重增加缓慢、糖水消耗减少、旷场试验各项指标较正常组间差异具有统计学意义;荧光定量PCR结果显示:模型组较对照组各脑区BDNF及Trkb受体表达减少,药物干预组脑区BDNF及Trkb受体表达显著高于模型对照组。结论:脑部BDNF因子及Trkb的表达在抑郁症发生与治疗中有所改变。