乌头碱中毒大鼠心肌酶的细胞化学研究——线粒体琥珀酸脱氢酶和细胞色素C氧化酶的超微结构定位

<正> 本文用细胞化学手段,对乌头碱中毒大鼠心肌线粒体的琥珀酸脱氢酶(SDH)、细胞色素C氧化酶(CCO)作超微结构定位,在电子显微镜下直接观乌头碱中毒后SDH、CCO的定位和活性的变化,发现乌头碱抑制了心肌大多数线粒体的SDH活性,并引起心肌线粒体CCO     

大鼠乌头碱中毒心肌酶的细胞化学研究(Ⅱ)——线粒体细胞色素C氧化酶(CCO)的定位

作者通过大鼠乌头碱中毒心肌线粒体 CCO 的超微结构定位,观察到乌头碱引起 CCO 的定位紊乱。由于损害程度不同,出现三种情况:(1)CCO 失去在线粒体嵴膜和内膜上的定位,酶反应产物呈自由扩散状态,酶反应强度对照组有所增强或相似;(2)CCO 失去在线粒体嵴膜和内膜上的定位,酶反应强度较对照组明显减弱;(3)在同一线粒体中部分保持 CCO 的嵴膜、内膜定位,部分失去 CCO 在嵴膜和内膜上的定位。实验结果表明,乌头碱干扰了心肌细胞呼吸链的电子传递,抑制氧化磷酸化的正常进行,导致心肌能量代谢障碍。     

大鼠乌头碱中毒心肌酶的研究(Ⅲ)——线粒体NADHD的组织化学和细胞化学研究

本文用组织化学和细胞化学方法分别对乌头碱染毒后不同时间组大鼠心肌NADHD进行了光镜和电镜观察。结果表明:乌头碱染毒后30分钟,2小时和4小时的大鼠心肌NADHD活性明显降低并有定位改变,而染毒后12、24小时的心肌NADHD活性有所回升。证实产能较多的NADH氧化呼吸链在乌头碱中毒时受到抑制。     

LC-MS~n分析新乌头碱在兔尿液中的代谢产物

乌头属(aconitum)中药临床应用广泛,治疗风湿痹痛等症疗效显著。但该属中药所含乌头类生物碱既是有效成分又是剧毒成分,其治疗量与中毒、致死量接近[1],因此服用过量、误服或投毒等都可引起乌头类生物碱中毒[2,3]。乌头碱中毒无特异性临床表现,且在体内迅速分解,给检测带来困难。在乌头属中药中新乌头碱(mesaconitine)多与乌头碱(aconi-tine)共存,且新乌头碱含量高于乌头碱。本文应用LC-MSn分析家兔灌服新乌头碱中毒死亡前尿液中的代谢产物,为检测新乌头碱的代谢产物提供新方法。1材料与方法1.1仪器与材料Finnigan公司LCQ液相色谱-质谱…     

大鼠乌头碱中毒心肌酶的研究(Ⅳ)——乳酸脱氢酶的组织化学和细胞化学定位

本文用组织化学和细胞化学相结合的方法,对乌头碱染毒后不同时间的大鼠心肌乳酸脱氢酶(LDH)进行了显微和超微结构定位。染毒后30min、2h 和4h,大鼠心肌 LDH 酶反应增强,至染毒后12h,24h 活性仍高于对照组。表明在有氧氧化障碍的同时,中毒鼠的心肌糖酵解增强,为乌头碱造成心肌能量代谢障碍的中毒机理进一步提供了证据。     

大鼠博落回总碱中毒后心肌细胞Bcl-2、Bax蛋白表达

目的 观察大鼠博落回总碱中毒后不同时间段心肌细胞内Bcl-2和Bax蛋白表达情况,为博落回中毒鉴定提供分子生物学标志物. 方法 制作大鼠博落回总碱中毒实验模型,通过免疫组化染色检测技术,观测不同时间段大鼠心肌细胞内Bcl-2和Bax蛋白表达情况,并对其结果 进行图像分析. 结果 博落回总碱中毒后不同时间段心肌细胞内Bcl-2和Bax蛋白表达不同,高于正常对照组(P<0.05).结论 博落回总碱中毒症状不明显情况下,应用免疫组织化学技术检测Bcl-2和Bax蛋白表达,可作为鉴定博落回总碱中毒的辅助手段.     

大鼠博落回总碱中毒心肌细胞凋亡的研究

目的观察博落回总碱中毒后大鼠不同时间内心肌细胞凋亡情况,为博落回总碱中毒提供毒理学指标。方法以SD大鼠博落回总碱中毒为实验模型,通过HE染色及凋亡细胞检测技术对大鼠博落回总碱中毒后心肌病理变化进行研究,并对研究结果进行计算机病理学图像分析。结果大鼠在中毒后的不同时间段内,心肌细胞凋亡数高于正常对照组(P<0.01),且凋亡数在不同时间段差异有显著性。结论博落回总碱中毒在临床症状不明显及毒物分析难以检测的情况下,应用凋亡细胞检测技术可成功检测出心肌的病理学变化。     

短柄乌头急性中毒——乌头碱在大鼠体内分布的研究

本文用高效液相色谱法检测了大白鼠短柄乌头急性中毒后(LD_(50)剂量灌胃),乌头碱在心、肝、肾、血、脑内的含量分布。结果表明;体内乌头碱含量甚微或检不出。30例大鼠LD_(50)剂量灌胃后,16例2h内中毒死亡。其肝、肾内的乌头碱检出率明显高于心、血、脑,且中毒表现明显。另14例于2h整处死,其肝、肾、血中检出较高。16例2h内死亡组肝中乌头碱含量分析提示乌头碱在体内代谢很快。此结果为法医工作中乌头碱中毒案件调查、检材提取、毒物分析结果的评价提供了一定的依据。     

乙醇-乌头碱致心律失常的毒理分子机制研究进展

乌头属植物是广泛使用的中草药代表之一,乌头碱是其中最主要的毒性成分,主要引起多种心律失常而致死。急性乙醇摄入也可引起心律失常。法医学实践中常遇到乌头类药酒引起的中毒案例。这两者的心肌毒性分子机制有较多共通点,均对心肌细胞膜钠通道、钙通道及钾通道等产生影响。本文通过综述近年来乙醇、乌头碱心脏毒性机制的研究报道,分析讨论乙醇-乌头碱可能共同影响的心肌细胞通道蛋白,为联合中毒致心律失常的分子机制研究提供思路与参考。     

原发性心肌病猝死心肌mtDNA~(4977)缺失变化

目的探讨原发性心肌病(PCM)猝死者心肌线粒体DNA(m tDNA4977)缺失情况及其与猝死的关系。方法对18例PCM猝死和28例对照组病例心肌组织蜡块,用常规方法提取心肌m tDNA,以PCR、琼脂糖紫外凝胶成像技术确定扩增产物激光密度,初步定量检测m tDNA4977缺失率。结果PCM猝死18例中,检见13例m tDNA4977缺失,占72.44%。对照组28例中,检见3例m tDNA4977缺失,占10.71%;两组病例m tDNA4977缺失率均值分别为0.5795和0.0744,差异有非常显著性意义。结论多数PCM,特别是扩张型心肌病猝死者心肌可检见m tDNA4977缺失;提示其心肌m tDNA4977缺失变化与PCM猝死的发生可能有一定关系。     

兔急性心肌缺血脱氢酶电镜—细胞化学变化的动态观察

为了研究急性心肌缺血脱氢酶变化的规律性，本文应用电镜酶细胞化学技术，通过结扎５０只家兔冠状动脉左前降支，观察心肌缺血后０．２５、０．５、１、１．５、２、４、６、１２、２４、３６、７２小时ＳＤＨ和ＬＤＨ的变化。结果显示：心肌缺血０．５小时后，ＳＤＨ和ＬＤＨ活性开始下降。同时伴有致密体的形成和超微结构的改变。随着缺血时间的延长，酶活性下降更加明显，最后消失。     

Isolated viable rat cardiac myocytes as a tool for evaluating histochemical methods for diagnosis of myocardial ischemia. I. Mitochondria-bound oxidoreductases — A preliminary report

A pilot study has been performed to evaluate the usefulness of isolated viable rat cardiac myocytes in the evaluation of histochemical oxidoreductase methods for estimating cellular anoxic damage. Data are presented in support of the use of succinate dehydrogenase and β-hydroxybutyrate dehydrogenase for evaluating different degrees of cellular damage. It appears that upon anoxia biochemical signs of membrane leakage do occur earlier than positive histochemical reactions. However, trypan blue staining occurs at a later time point. Probably the histochemical reaction-rates are limited by the penetration of both the substrate and the electron acceptor, in this case nitro blue tetrazolium, into the cells and the mitochondria.     

Validation of Ultrastructural Analysis of Mitochondrial Deposits in Cardiomyocytes as a Method of Detecting Early Acute Myocardial Infarction in Humans*

Abstract: In the present study, ultrastructural analysis of mitochondrial deposits (black dots within mitochondria) as a method for the detection of early acute myocardial infarction (AMI) was evaluated. In 24 patients with AMI and six controls, analysis was performed in the heart of infarcted patients and noninfarcted controls. In the infarction area in lactate dehydrogenase (LDH)‐diagnosed AMI, the percentage of positive mitochondria was significantly higher compared to corresponding heart tissue in control patients and compared to noninfarcted areas within these patients. Also in patients with a clinically diagnosed AMI but no LDH decoloration, a significant higher percentage of positive mitochondria was found in the left ventricle compared to controls and noninfarcted areas. In patients with AMI, an increase in mitochondria with deposits was found in the infarction area compared to controls and noninfarcted tissue within the same patient, suggesting that electron microscopical changes in mitochondria can be used for the diagnosis of AMI less than 3 h old.     

Oxidative enzyme activities and respective histochemical reactions in ischemic rat myocardium

To evaluate the diagnostic role of histochemically demonstrated aerobic dehydrogenases in ischemic myocardial injury NADH-diaphorase, succinate dehydrogenase (SDH), beta-hydroxybutyrate dehydrogenase (HBDH) and malate dehydrogenase (MDH) were demonstrated histochemically and the corresponding enzyme activities were measured biochemically in isolated perfused rats hearts after global ischemia from 0 to 12 h. The present data show that the enzyme-histochemical methods when used properly are more sensitive indicators of early ischemic injury than classical histological staining procedures. From the enzymes tested here the histochemical demonstration of HBDH turned out to be best suited for use when suspecting ischemic myocardial injury at autopsy.     

早期心肌挫伤超微结构变化的实验研究

应用机械式弹性拉力打击器以10.0米／秒速度冲击犬胸骨心前区，引起心肌挫伤。分别对伤后2～5min、30～60min、3h及5h心肌挫伤区超微结构变化进行观察。结果显示：（1）伤后2～5min即见严重的心肌超微结构损害，如肌原纤维撕裂、扭曲变形，肌节过度收缩或I带增宽Z线弯曲、线粒体破裂，部分嵴不清、间质出血并可见游离的线粒体。伤后30～60分钟挫伤心肌肌原纤维溶解、碎裂，线粒体破裂伴嵴消失。伤后3及5h，挫伤区坏死明显，坏死物与红细胞碎片混杂一起，结构不能辨认。（2）同一心脏可有多部位损伤，且损伤程度不均一。     

显示和判定心肌早期病变的染色方法研究

作者用探讨的变色酸2R亮绿法和其他10余种染色方法,对心血管病猝死的尸检心肌和经心电、Mb、CPK、LDH、SOD诊断有急性心肌损伤的多种大量动物实验材料观察结果,证明心肌早期病变有染色性质异常,对显示和判定心肌早期病变有实用价值。提出并讨论了心肌早期病变及其显示、判定方法。     

家兔心肌自溶蛋白质降解与死亡时间的关系

本文对家兔心肌自溶时的蛋白质水解和氨基酸代谢进行了研究。心肌组织在37℃湿盒中自溶15～480min,用离子交换色谱法测定游离氨基酸以及组织氨含量。自溶时大多数组织游离氨基酸含量上升,丙氨酸、赖氨酸和组氨酸含量在自溶15min即有明显增加,而谷氨酸含量呈下降趋势,同时组织氨含量上升。结果提示:心肌自溶早期蛋白质降解加速。上述结果有助于死亡时间的推断。     

垂体后叶素致冠状动脉痉挛症的实验病理学研究

作者通过大鼠舌下静脉注射垂体后叶素致冠状动脉痉挛,运用组织学、酶组织化学及电镜技术研究冠状动脉痉挛症引起心肌急性缺血的形态学变化特点,同时对大鼠心肌自溶性变化与上述结果进行了分析比较,认为心肌纤维磷钨酸苏木素(PTAH)染色出现不规则横带且伴有心肌间质小血管壁腺苷三磷酸酶(ATPase)活性下降,有助于冠状动脉痉挛症引起心肌缺血的诊断。     

EXPLAINING THE GENDER GAP IN CRIME: THE ROLE OF HEART RATE

Although it is well established that males engage in more crime compared with females, little is known about what accounts for the gender gap. Few studies have been aimed at empirically examining mediators of the gender–crime relationship in a longitudinal context. In this study, we test the hypothesis that a low resting heart rate partly mediates the relationship between gender and crime. In a sample of 894 participants, the resting heart rate at 11 years of age was examined alongside self‐reported and official conviction records for overall criminal offending, violence, serious violence, and drug‐related crime at 23 years of age. A low resting heart rate partially mediated the relationship between gender and all types of adult criminal offending, including violent and nonviolent crime. The mediation effects were significant after controlling for body mass index, race, social adversity, and activity level. Resting heart rate accounted for 5.4 percent to 17.1 percent of the gender difference in crime. This study is the first to produce results documenting that lower heart rates in males partly explain their higher levels of offending. Our findings complement traditional theoretical accounts of the gender gap and have implications for the advancement of integrative criminological theory.