生长分化因子5真核表达质粒转染小鼠骨髓基质干细胞体外诱导向软骨细胞分化

背景:生长分化因子5是软骨与骨组织形成、发育的重要调解因子,在诱导软骨形成,促进骨、软骨、肌健韧带损伤修复方面发挥重要作用.目的:小鼠骨髓基质干细胞体外转染真核表达质粒pcDNA 3.1(+),生长分化因子5,检测与软骨形成分化相关的细胞外基质及蛋白多糖的表达.设计、时间及地点:细胞学体外观察,于2008-03/12在武汉协和医院中心实验室完成.材料:雄性昆明种小鼠20只,由华中科技大学同济医学院实验动物中心提供.生长分化因子5真核表达质粒pCDNA 3.1(+)/生长分化因子5为自备.方法:全骨髓贴壁法体外分离培养小鼠骨髓基质干细胞,取传至第3代细胞接种到6孔板,在细胞生长到90%融合时开始转染.设立3组:转染组采用Lipofectamine<'TM>2000进行脂质体介导pcDNA3.1(+)/生长分化因子5重组质粒瞬时转染;空质粒组转染窄质粒pcDNA 3.1(+);空白对照组只加入等量脂质体,其余步骤相同.主要观察指标:转染后72 h通过RT-PCR及免疫细胞化学检测生长分化因子5基因与蛋白的表达鉴定转染是否成功,同法检测软骨基质Ⅱ型胶原的表达,转染后14 d阿尔辛蓝染色检测蛋白聚糖的表达.结果:转染组有一大小为219 bp的特异性扩增条带,骨髓基质干细胞胞浆内呈棕色阳性染色;而空质粒组、空白对照组均未发生生长分化因子5基因转录,无特异性扩增条带,且细胞胞浆未见明显染色.转染组町检测到Ⅱ型胶原基因的表达,基因大小225 bp,Ⅱ型胶原细胞胞浆中可见棕黄色染色;空质粒组、空白对照组均未检测到Ⅱ型胶原基因的表达,SP染色均无明显染色.阿尔辛蓝染色后转染组细胞呈蓝染,空质粒组、空白对照组均未见明显异染性着色.结论:pcDNA3.1(+)/生长分化因子5转染骨髓基质干细胞能显著增加Ⅱ型胶原及蛋白聚糖的表达,促进骨髓基质干细胞向软骨细胞方向分化.     

补肾壮骨颗粒干预维甲酸致骨质疏松大鼠的骨生物力学变化

背景:以中药复合成分制成的补肾壮骨颗粒应用于临床治疗骨质疏松症已十余年.目的:验证补肾壮骨颗粒对维甲酸致骨质疏松大鼠骨生物力学的影响.设计、时间及地点:随机对照动物实验,于2008-01/06广州医药工业研究所完成.材料:3月龄SD大鼠60只,雌雄各半,体质量227～369 g.按体质量随机分为5组:正常对照组、骨质疏松模型组、补肾壮骨颗粒治疗高、低剂量组、仙灵骨葆阳性对照组,每组12只.补肾壮骨颗粒主要由鹿角胶、龟胶、山药、水蛭等药物组成;仙灵骨葆胶囊由贵州同济堂制药有限公司提供;维甲酸由武汉远城科技发展有限公司提供.方法;各组大鼠上午均灌胃给予维甲酸2周,造模同时下午按指定剂量灌胃给予相应药物:补肾壮骨颗粒低剂量组2.5g/kg,高剂量组5.0 g/kg;仙灵骨葆阳性对照组1.1 g/kg;正常对照组以及骨质疏松模型组给予等量的蒸馏水,1次/d,连用3周.麻醉后处死动物进行大鼠左侧股骨3点弯曲试验、左侧胫骨3点弯曲试验、第4腰椎骨压碎试验.主要观察指标:各组大鼠左侧股骨最大荷载、第4腰椎骨骨结构强度及左侧胫骨骨结构强度.结果:模型组大鼠的股骨最大荷载、椎骨结构强度均显著低于正常,表明给大鼠灌胃维甲酸,可导致大鼠骨生物力学性能的降低;与模型组相比,各给药组的左侧股骨最大荷载、第4腰椎结构强度、左侧胫骨弯曲试验的最大荷载结果差异均无显著性意义,但均有增加趋势,其中补肾壮骨颗粒低剂量组的左股骨弯曲试验的最大荷载、左胫骨弯曲试验的最大荷载、第4腰椎压碎试验的结构强度分别增加了16.2%,17.7%,15.4%.结论:补肾壮骨颗粒能增加小梁骨的骨强度,以低剂量组效果最佳,起到防治骨质疏松症的作用.     

人性化护理在初产妇自然分娩中的应用效果观察

[目的]观察人性化护理在初产妇自然分娩中的应用效果.[方法]218例住院正常足月待产的初产妇随机分为观察组93例和对照组125例,对照组给予常规产科护理,观察组采取人性化护理.观察并比较两组产妇分娩方式、自然分娩产妇疼痛程度和自然分娩产妇各产程时间.[结果]与对照组比较,观察组产妇分娩疼痛程度减轻(P<0.05);剖宫产率下降,自然分娩率升高(P<0.05);产程缩短.[结论]人性化护理应用于初产妇自然分娩中效果明显.     

PDCA工作循环理论在2型糖尿病病人健康教育中的应用

[目的]探讨质量管理工作循环(PDCA)理论在2型糖尿病病人健康教育中的应用.[方法] 368例2型糖尿病病人随机分为两组,观察组198例和对照组170例,分别在健康教育前及健康教育7个月后进行糖尿病相关知识认知情况和病人对健康教育的满意度情况问卷调查.[结果] 265例病人完成随访,观察组140例,对照组125例;观察组健康教育后糖尿病相关知识认知情况和病人对健康教育的满意度情况均优于对照组.[结论]在2型糖尿病病人健康教育中应用PDCA工作循环理论效果明显.     

钛表面粗糙度对牙周韧带细胞早期附着影响的荧光观察

背景:种植体表面粗糙程度对组织细胞的增殖、分化及基因表达有直接影响.目的:观察牙周韧带细胞在不同纯钛表面粗糙度的早期附状态,以及材料表面性能对组织细胞分化的影响.设计、时间及地点:随机对照观察,多样本比较实验,于2005-01/2006-07在赣南医学院科研中心完成.材料:采用机床切割机将商业纯钛棒制备成直径为10mm厚2mm的钛片,共24个,分为4组:机械处理组,硝酸处理组,喷砂处理组,综合处理组,每组6个样品.方法:采用TR240便携式表面粗糙度仪测定表面粗糙度.机械处理组:进行单纯的机械处理.硝酸处理组:单纯的机械处理+65%的硝酸(100℃,1 h)处理.喷砂处理组:单纯的机械处理+100 μ m的Al2O3喷砂处理.综合处理组:单纯的机械处理+100 μ m的Al2O3喷砂处理后+65%的硝酸(100℃,1 h)处理.主要观察指标:样品在DMEM中预置30 m后接种第3代的细胞,分别于30,60,120,240 min,1,3,7 d取出钛片,于荧光显微镜下观察各组纯钛表面的粗糙度和牙周韧带细胞在不同表面粗糙度的纯钛表面早期附着情况.结果:①定量分析:4组粗糙度分别为:机械处理组(599.5±8.3)nm、硝酸处理组(406.5±4.6)nm、喷砂处理组(358.8±11.8)nm、综合处理组(8.7±2.0)nm,机械处理组高于硝酸处理组、喷砂处理组和综合处理组(P<0.01),差异有显著性意义(P<0.01).②定性分析:牙周韧带细胞在纯钛表面早期附着的荧光观察显示,随着时间的推移,各组早期附着在纯钛表面的牙周韧带细胞增多,细胞增殖良好.但粗糙度大的钛表面,细胞附着较少,粗糙度小的钛表面,细胞附着较致密些.结论:钛表面粗糙度越小,牙周韧带细胞早期附着越多,有利于细胞黏附和增殖.     

山东省带绦虫病与囊虫病重点疫区流行现状调查研究

目的 了解山东省带绦虫病与囊虫病在重点疫区的流行情况。方法 １９９９年对聊城等４市地的５县（区）１５个乡（镇）４３个行政村的３１１２４人进行了调查,结果 人群带绦惠恨,囊虫感染率５．１／万,有较明显的职业分布特征。人群血清猪囊尾蚴特异性Ｉｇｇ,抗体阳性率１．３８％,商品猪血清囊尾蚴抗体阳性率０．３８％。结论 初步搞清了山东省重点疫区带绦虫病与囊虫病的流行现状,并对该病的各流行环节做了分析,为该病的     

Value of telomerase activity in the differential diagnosis of benign and malignant ascites

OBJECTIVE : To evaluate telomerase activity as a marker for the differentiation of benign and malignant ascites. METHODS : Exfoliated cells were harvested from various kinds of ascitic fluid and telomerase activity was determined by TRAP‐PCR‐ELISA (tartrate‐resistant acid phosphatase–polymerase chain reaction–enzyme‐linked immunosorbent assay). RESULTS : Telomerase activity could not be detected in the ascites of hepatic cirrhosis or peritoneal tuberculosis but was detected in 29 of 34 (85.2%) cancerous ascites. CONCLUSIONS : Telomerase activity may be present in cancerous ascites and could be used as a marker for the differential diagnosis of benign and malignant ascites.     

高血压合并2型糖尿病患者心率震荡的研究

目的探讨窦性心率震荡(HRT)、静息心率(RHR)和心率变异性(HRV)时阈指标(SDNN)在单纯高血压及高血压合并2型糖尿病中的变化及相互关系。方法选择单纯高血压46例、高血压合并2型糖尿病22例和健康人50例,分别为单纯高血压组、高血压合并2型糖尿病组和对照组。根据24h动态心电图记录,分别计算各组HRT的两个参数震荡初始值(TO)和震荡斜率值(TS)以及HRV的全部NN间期的标准差(SDNN)和6:00时的RHR,比较各组间的差别,分析TO、TS与SDNN、RHR之间的相关性。结果高血压合并2型糖尿病组TO、TS、TO+TS、SDNN的阳性检出率均高于单纯高血压组及对照组,组间比较差异有显著意义;RHR值明显高于单纯高血压组及对照组,组间比较差异有显著意义。高血压合并2型糖尿病组TO与SDNN呈负相关,与RHR呈正相关;TS与RHR呈负相关,而与SDNN呈正相关。结论高血压患者特别是合并有2型糖尿病者,HRT减弱及HRV异常,RHR增加,提示HRT及RHR可以作为早期发现高血压或合并2型糖尿病患者心脏自主神经功能紊乱的一个有效的可靠无创指标。     

Natural tri- to hexapeptides self-assemble in water to amyloid beta-type fiber aggregates by unexpected alpha-helical intermediate structures

Many fatal neurodegenerative diseases such as Alzheimer's, Parkinson, the prion-related diseases, and non-neurodegenerative disorders such as type II diabetes are characterized by abnormal amyloid fiber aggregates, suggesting a common mechanism of pathogenesis. We have discovered that a class of systematically designed natural tri- to hexapeptides with a characteristic sequential motif can simulate the process of fiber assembly and further condensation to amyloid fibrils, probably via unexpected dimeric α-helical intermediate structures. The characteristic sequence motif of the novel peptide class consists of an aliphatic amino acid tail of decreasing hydrophobicity capped by a polar head. To our knowledge, the investigated aliphatic tripeptides are the shortest ever reported naturally occurring amino acid sequence that can adopt α-helical structure and promote amyloid formation. We propose the stepwise assembly process to be associated with characteristic conformational changes from random coil to α-helical intermediates terminating in cross-β peptide structures. Circular dichroism and X-ray fiber diffraction analyses confirmed the concentration-dependent conformational changes of the peptides in water. Molecular dynamics simulating peptide behavior in water revealed monomer antiparallel pairing to dimer structures by complementary structural alignment that further aggregated and stably condensed into coiled fibers. The ultrasmall size and the dynamic facile assembly process make this novel peptide class an excellent model system for studying the mechanism of amyloidogenesis, its evolution and pathogenicity. The ability to modify the properties of the assembled structures under defined conditions will shed light on strategies to manipulate the pathogenic amyloid aggregates in order to prevent or control aggregate formation.     

Therapeutic options and heart failure survival score predictability in an academic heart failure center: an analysis of 120 consecutive patients during a 1-year period

BACKGROUND: the incidence and prevalence of patients with advanced heart failure is increasing worldwide and the number of cardiac transplantations remains limited. AIMS: it was the aim of the study to describe our experience with the increasing number of available medical, interventional and cardiac surgery options, and to assess heart failure survival score predictability in an academic heart failure center within a 1-year follow-up. METHODS AND RESULTS: in all patients who were referred for cardiac transplant evaluation within a 12-month period between April 1998 and March 1999 at our Interdisciplinary Heart Failure and Transplant Program, our team assessed all medical interventions as well as interventional and surgical treatment options that were available, based on the clinical profile on initial presentation. In 92% of the 120 patients referred for cardiac transplantation evaluation, drug therapy could be optimized. A considerable number of patients could be subjected to an organ-preserving intervention or surgery, either PTCA (n=11), CABG (n=4), valve repair (n=7), multisite pacing (n=7), or partial ventricular resection (n=5). Only a small group of patients with the worst heart failure survival score were listed for heart transplantation (n=17) or received a ventricular assist device (n=3). CONCLUSIONS: within a contemporary cohort of advanced heart failure patients, only a small number of patients will undergo cardiac transplantation, which is predictable by the heart failure survival score. Most patients will undergo optimized medical therapy and a considerable number will be subjected to interventional or surgical treatment options.