大黄素通过非Caspase依赖通路诱导膀胱癌细胞BIU87凋亡的机制

目的 研究大黄素对人膀胱癌细胞BIU87凋亡及凋亡诱导因子(apoptosis inducing factor,AIF)和核酸内切酶G( Endonuclease G,Endo G)的mRNA和蛋白表达的影响,探讨大黄素的抗癌机制. 方法 取对数生长期的人膀胱癌细胞株BIU87,分别加入大黄素和Caspase抑制剂Z-VAD-FMK共同培养.实验分为4组:空白对照组、Z-VAD-FMK组、大黄素组、大黄素+Z-VAD-FMK组.采用甲基噻唑(MTT)比色法测定不同浓度大黄素在不同作用时间内对体外培养的BIU87细胞增殖的影响,应用流式细胞仪检测细胞凋亡,RT-PCR检测各组细胞AIF和Endo G mRNA的表达,蛋白质印迹法检测各组细胞AIF和Endo G蛋白的表达. 结果 MTT检测显示,大黄素的浓度越高、作用时间越长,对肿瘤细胞生长的抑制作用越强.根据IC50值,选择80 μmol/L的大黄素干预72 h作为干预条件.大黄素组细胞凋亡率为(44.57±1.52)％,大黄素+Z-VAD-FMK组为(35.58±1.61)％,差异有统计学意义(P＜0.05).RT-PCR和蛋白质印迹结果显示,大黄素+Z-VAD-FMK组中AIF mRNA和蛋白为1.74±0.11、2.59±0.13,大黄素组为1.36±0.08、1.89±0.14,空白对照组为0.37±0.02、0.53±0.11,Z-VAD-FMK组为0.42±0.06、0.44 ±0.07,大黄素+Z-VAD-FMK组与其他各组比较,差异有统计学意义(P＜0.05);大黄素+ Z-VAD-FMK组中Endo G mRNA和蛋白为2.28±0.15、3.31±0.36,大黄素组为1.85±0.13、2.15 ±0.27,空白对照组为0.53±0.07、0.71 0.16,Z-VAD-FMK组为0.61±0.04、0.67±0.22,大黄素+Z-VAD-FMK组与其他各组比较,差异有统计学意义(P＜0.05). 结论 大黄素可上调膀胱癌细胞BIU87中AIF和Endo G的表达,通过启动非Caspase依赖途径诱导细胞凋亡.     

易-和康复技术改善缺血性中风痉挛性偏瘫随机对照研究

目的 观察易-和康复技术改善缺血性中风后痉挛性偏瘫的临床疗效.方法 将171例缺血性中风后痉挛性偏瘫的患者,通过中央随机系统按2:1比例将患者随机分为治疗组114例、对照组57例.其中,治疗组脱落17例,完成病例观察97例,脱落率为14.9％;对照组脱落8例,完成病例观察49例,脱落率为14.0％.治疗组采用易-和康复...     

对"异病同治"的认识

"异病同治"是中医学的特色,体现了中医辨证论治的精神。本文试图从辨证论治和辨病论治相结合,不同疾病可存在相同或相似的病机变化,和指导病证结合横向研究三方面加强对"异病同治"的认识。     

乳腺空芯针活检为不典型导管上皮增生的病理低估研究

目的:计算乳腺空芯针穿刺活检(core needle biopsy,CNB)诊断为不典型导管上皮增生(atypical ductal hyperplasia,ADH)病人的病理低估率。分析病人临床及影像学信息,探讨低估的预测因素。方法:回顾性分析2010年1月至2013年2月期间,本中心60例CNB诊断为ADH病人。定义病理低估为CNB是ADH但切除活检诊断是恶性的。以卡方检验、Fisher精确检验和二分类Logistic回归分析病理低估的预测因素。结果:本研究60例CNB诊断为ADH病人的病理低估率为65.0%(39/60)。乳腺X线检查发现恶性征象的微钙化(OR=7.988,95%CI:4.997~12.810,P=0.001)、乳腺X线检查影像报告数据系统(BI-RADS)≥4级(OR=10.875,95%CI:2.747~43.051,P     

改良留置导尿管相关尿路感染病原菌及感染危险因素研究

目的 研究改良留置导尿管相关尿路感染患者病原菌的分布情况,分析改良留置导尿管患者尿路感染的危险因素。方法 纳入2018年3月至2020年3月邯郸市第一医院收治的168例改良留置导尿管患者作为研究对象,观察所有患者尿路感染和病原菌分布情况。将发生尿路感染和未发生尿路感染的患者收集资料并进行对比,差异有统计学意义项目代入Logistic回归方程计算,分析改良留置导尿管患者发生尿路感染的危险因素。结果 168例改良留置导尿管患者中有49例患者发生尿路感染,发生率为29.17%。共检出病原菌84株。其中革兰阴性杆菌50株（59.52%）,革兰阳性球菌24株（28.57%）,真菌10株（11.90%）。发生尿路感染和未发生尿路感染患者在年龄、住院时间、留置导尿管时间、糖尿病、肾结石、低蛋白血症以及抗菌药物使用种类上对比差异均有统计学意义（P<0.05）。将上述项目代入Logistic回归方程计算发现,年龄≥60岁、住院时间≥21 d、留置导尿管时间≥13 d、合并糖尿病、合并肾结石、合并低蛋白血症以及尿路感染前抗菌药物使用种类≥3种均是改良留置导尿管患者发生尿路感染的危险因素。结论 改良留...     

Application of the assay of urine FSH β-subunit in patients with pubertal disorders

Objective: To evaluate the diagnostic predictive value in the identification of puberty disorders by means of ELISA of β-FSH subunit levels in successively collected urine samples compared to RIA of intact FSH in serum obtained from the normal control subjects and patients with puberty disorders, respectively.Subjects and Methods: Five male and four female volunteers were recruited as controls. Four patients with the hypogonadotropic hypogonadism, five patients with hypergonadotropic hypogonadism, four patients with the central precocious puberty and one patient with isosexual peripheral precocious puberty collected successively their early-morning urine samples for 30 to 32 days.The urine β-FSH subunit was assayed with the method of ELISA, then adjusted by creatinine (Cr) concentration.Results:Comparing with their cotemporary groups, patients with the hypogonadotropic hypogonadism had lower levels of urine β-FSH, and patients with idiopathic hypergonadism had higher levels with irregular fluctuation. Meanwhile, patients with the central precocious puberty had much higher levels of urine β-FSH with irregular peaks, and patients with isosexual peripheral precocious puberty had almost normal levels. The patterns were coincident with the clinical characteristics and serum FSH levels.Conclusion: The ELISA of urine β-FSH subunit possesses a number of advantages over the RIA of serum FSH level, such as low cost, simplicity and reliability in the clinical practice. It can be used for the diagnoses of puberty disorders. In addition, it is possible and much easier, comparing with blood samples, to collect successively urine samples for research of pathophysiological dynamics of FSH secretion in puberty disorders and other reproductive dysfunction.     

The use of 7-amino actinomycin D in identifying apoptosis: simplicity of use and broad spectrum of application compared with other techniques

The detection and quantitation of apoptotic cells is becoming increasingly important in the investigation of the role of apoptosis in cellular proliferation and differentiation. The pathogenesis of hematologic disorders such as aplastic anemia and the development of neoplasia are believed to involve dysregulation of apoptosis. To quantitate accurately the proportion of apoptosis cells within different cell types of a heterogeneous cell population such as blood or bone marrow, a method is required that combines the analysis of large numbers of cells with concurrent immunophenotyping of cell surface antigens. In this study, we have evaluated such a method using the fluorescent DNA binding agent, 7-amino actinomycin D (7AAD), to stain three diverse human cell lines, induced to undergo apoptosis by three different stimuli. Flow cytometric analysis defines three populations on the basis of 7AAD fluorescence and forward light scatter. We have shown by cell sorting and subsequent morphological assessment and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling that the populations defined by 7AAD represent live, apoptotic, and late-apoptotic/dead cells. This method is quick, simple, reproducible, and cheap and will be a valuable tool in the investigation of the role of apoptosis in normal physiology and in disease states.     

结核分枝杆菌Rv1009基因蛋白的克隆和表达

目的克隆表达结核分枝杆菌促Rv1009基因,序列测定正确后进行融合、表达。方法采用热启动聚合酶链反应(Polymerase Chain Reaction,PCR)从结核分枝杆菌H37Rv基因组中扩增出Rv1009编码基因,用限制性内切酶消化后插入pGEX 4T-2载体中,将重组质粒转化大肠杆菌BL21 (DE3),目的基因经IPTG诱导,表达Rv1009基因蛋白。结果经PCR扩增在1300bp处发现一条目的片段,获得了结核分枝杆菌H37Rv株Rv1009基因蛋白,经诱导后高效表达分子量为64KD的外源蛋白,与预期分子量大小一致,凝胶自动扫描分析,在A600值为0．6,IPTG终浓度为0．3 mmol／L,诱导表达3 h时融合蛋白表达量即达峰值,占菌体总蛋白的22．8％。结论构建了结核分枝杆菌Rv1009基因重组表达载体,获得了RPF样融合蛋白的高效表达,为今后深入研究奠定了基础。     

Fe/Co/N–C/graphene derived from Fe/ZIF-67/graphene oxide three dimensional frameworks as a remarkably efficient and stable catalyst for the oxygen reduction reaction

The development of non-noble metal catalysts with high-performance, long stability and low-cost is of great importance for fuel cells, to promote the oxygen reduction reaction (ORR). Herein, Fe/Co/N–C/graphene composites were easily prepared by using Fe/ZIF-67 loaded on graphene oxide (GO). The Fe/Co/porous carbon nanoparticles were uniformly dispersed on graphene with high specific surface area and large porosity, which endow high nitrogen doping and many more active sites with better ORR performance than the commercial 20 wt% Pt/C. Therefore, Fe/Co/N–C/graphene composites exhibited excellent ORR activity in alkaline media, with higher initial potential (0.91 V) and four electron process. They also showed remarkable long-term catalytic stability with 96.5% current retention after 12 000 s, and outstanding methanol resistance, compared with that of 20 wt% Pt/C catalysts. This work provides an effective strategy for the preparation of non-noble metal-based catalysts, which could have significant potential applications, such as in lithium–air batteries and water-splitting devices.

Fe/Co/N–C/graphene was facilely and successfully prepared by a calcination process, which has remarkable electrocatalytic ORR activity in alkali solutions and also displays an exceptional stability for the ORR and methanol tolerance.