优化的热启动PCR—限制性酶切法检测冠心病患者？…

改进传统的载脂蛋白Ｅ基因多态性检测方法，建立热启动聚合酶链反应－限制性片段长度多态性检测分析方法。方法：应用热启动ＰＣＲ－限制性内切酶酶切－聚丙烯酰胺凝胶电泳－固定银染法测定１５８例冠心病患者和１１６例正常对照者的ＡｐｏＥ基因型。     

街头篮球赛改善血液载脂蛋白水平的运动处方研究

通过街头篮球赛方式检验其作为改善血液载脂蛋白水平的有效性;受试者每周4次,每次4×10min进行中等强度街头篮球赛,18周后检测受试者血液载脂蛋白水平(ApoA1,ApoB及ApoA1/ApoB)和体型特征数据并与受试前比较;经过18周的街头比赛,受训者多数的血液载脂蛋白水平得到改善,说明街头篮球赛可作为改善血液载脂蛋白水平的一个运动处方.     

Molecular basis of Alzheimer's disease

Despite an exponential production of data, Alzheimer's disease (AD) remains an enigma. Unresolved questions persist in the face of the heterogeneity of this neuropathology. Recent progress in understanding mechanisms for AD results from the study of amyloid precursor protein (APP) metabolism and the involvement of senile plaque-associated proteins. In addition to the amyloid cascade hypothesis, alternative schemes emerge, in which the amyloid peptide is not the primary effector of the disease. Perturbations of vesicular trafficking, the cytoskeletal network, and membrane cholesterol distribution could be central events. Furthermore, since the physiological role of APP, presenilins, and apolipoprotein E in the central nervous system are not completely understood, their involvement in AD etiology remains speculative. New actors have to be found to try to explain sporadic cases and non-elucidated familial cases.     

Low-density lipoprotein receptor-mediated endocytosis of PEGylated nanoparticles in rat brain endothelial cells

Poly(methoxypolyethyleneglycol cyanoacrylate-co-hexadecylcyanoacrylate) (PEG-PHDCA) nanoparticles have demonstrated their capacity to diffuse through the blood-brain barrier after intravenous administration. However, the mechanism of transport of these nanoparticles into brain has not yet been clearly elucidated. The development of a model of rat brain endothelial cells (RBEC) in culture has allowed investigations into this mechanism. A study of the intracellular trafficking of nanoparticles by cell fractionation and confocal microscopy showed that nanoparticles are internalized by the endocytic pathway. Inhibition of the caveolae-mediated pathway by preincubation with filipin and nystatin did not modify the cellular uptake of the nanoparticles. In contrast, chlorpromazine and NaN₃ pretreatment, which interferes with clathrin and energy-dependent endocytosis, caused a significant decrease of nanoparticle internalization. Furthermore, cellular uptake experiments with nanoparticles preincubated with apolipoprotein E and blocking of low-density lipoprotein receptors (LDLR) clearly suggested that the LDLR-mediated pathway was involved in the endocytosis of PEGPHDCA nanoparticles by RBEC. Received 1 September 2006; received after revision 4 December 2006; accepted 18 December 2006     

Interaction of Apolipoprotein H with Phospholipid Membranes : a Monolayer Study

ＩｎｔｒｏｄｕｃｔｉｏｎＡｐｏｌｉｐｏｐｒｏｔｅｉｎＨ（ＡｐｏＨ）ｉｓａｐｌａｓｍａｇｌｙｃｏｐｒｏｔｅｉｎｂｅｌｏｎｇｉｎｇｔｏｔｈｅｆａｍｉｌｙｏｆａｐｏｌｉｐｏｐｒｏｔｅｉｎｓ．Ｉｔｗａｓｄｅｓｃｒｉｂｅｄｆｏｒｔｈｅｆｉｒｓｔｔｉｍｅｉｎ１９...     

PCR 技术用于 DNA 多态性分析 apoB—3′HVR 和 D17S30位点的体外扩增

本文用PCR技术扩增了北京地区73名无关个体的apoB—3'HVR和其中50名的D17S30位点.经琼脂糖凝胶电泳检测,在人群中表现出一定程度的多态性;同一样品分别扩增apoB—3'HVR和D17S30,其扩增带有明显的差别;而同一样品分次扩增后,其扩增带完全一致,表明扩增产物的可重复性.与DNA 指纹技术相比,这种方法虽然鉴别能力较低,但操作简便,灵敏度高,适于微量生物样品检验.我们将该技术应用于两例亲子鉴定案,其中一例排除了嫌疑父亲,另一例则不能排除.     

低密度脂蛋白胆固醇、载脂蛋白B与颈动脉粥样硬化斑块的关系

目的 :观察颈动脉粥样硬化患者颈动脉的结构变化及粥样硬化斑块情况 ,研究低密度脂蛋白胆固醇 (L DL - C)、载脂蛋白 B(Apo B)与颈动脉粥样硬化斑块的关系。方法 :测定 10 0例患者空腹血中 L DL - C、Apo B浓度 ,并行颈动脉超声检查 ,测量斑块面积 ,并计算斑块积分和斑块指数。分别计算 L DL - C、Apo B与颈动脉粥样硬化斑块总积分的相关性。按 L DL - C/ Apo B比值分组 ,计算各组的平均斑块指数。结果 :L DL - C、Apo B与颈动脉粥样硬化斑块总积分之间均呈正相关 ,且二者的相关性相当 (r=0 .6 6 5 7,P<0 .0 1;r=0 .6 86 4 ,P<0 .0 1)。L DL - C/ Apo B<1.2 5组颈动脉粥样硬化斑块指数比 L DL - C/ Apo B>1.6 2组高 ,两者相比有显著性差异 (1.76± 0 .80 ,3.30± 1.4 6 ;P<0 .0 5 )。结论 :L DL - C、Apo B与颈动脉粥样硬化斑块的形成均有密切关系 ,其中 Apo B与颈动脉粥样硬化斑块的关系更密切     

血脂检测预测冠状动脉病学的临床意义

目的通过检测血脂在冠状动脉病变中的变化,了解血脂分析预测冠状动脉病变的临 床意义。方法设置病变组和对照组,检测胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(RDL)、载脂 蛋白AI(APOAI)、载脂蛋白B(APOB)、脂蛋白a[Lp(a)]。结果病变组APOAI、APOB与对照组比较 具有高度显著性差异(P《0．001);TC、TG、HDL、Lp(a)次之(P<0．01);各项指标检测异常比例以 APOAI、APOB、LP(a)最高(69．5％、72.O％、65．9％)。结论血脂6项指标对预测冠状动脉病变均有 临床意义,建议这6项指标均应成为冠状动脉病变预测的常规检测项目。     

Cell surface adenylate kinase activity regulates the F1-ATPase/P2Y13-mediated HDL endocytosis pathway on human hepatocytes

We have previously demonstrated on human hepatocytes that apolipoprotein A-I binding to an ecto-F₁-ATPase stimulates the production of extracellular ADP that activates a P2Y₁₃-mediated high-density lipoprotein (HDL) endocytosis pathway. Therefore, we investigated the mechanisms controlling the extracellular ATP/ADP level in hepatic cell lines and primary cultures to determine their impact on HDL endocytosis. Here we show that addition of ADP to the cell culture medium induced extracellular ATP production that was due to adenylate kinase and nucleoside diphosphokinase activities, but not to ATP synthase activity. We further observed that in vitro modulation of both ecto-NDPK and AK activities could regulate the ADP-dependent HDL endocytosis. But interestingly, only AK appeared to naturally participate in the pathway by consuming the ADP generated by the ecto-F₁-ATPase. Thus controlling the extracellular ADP level is a potential target for reverse cholesterol transport regulation. Received 13 July 2006; received after revision 29 August 2006; accepted 19 September 2006     

二甲亚砜在聚合酶链反应扩增载脂蛋白E基因中的作用

目的：探讨二甲亚砜（DMSO）在聚合酶链反应（PCR）中扩增人类载脂蛋白E基因中的作用。方法：在不同质量分数DMSO的存在下,以PCR技术增人类载脂蛋白E基因片段。限制性内切酶酶切检测扩增产物。结果：在质量在分数5%-10%的DMSO下,可以成功地扩增到人类载脂蛋白E基因的第4外显子片段。结论：在DMSO作用下,PCR扩增增强了特异性。扩增的人类载脂蛋白E基因片段可以进行基因限制性片段多态性的分析。