柯萨奇病毒和腺病毒受体在VMC和DCM猝死者心肌组织中的表达

目的探讨病毒性心肌炎(VMC)和扩张性心肌病(DCM)的发病机制及相互关系。方法应用改良免疫组化技术检测病毒性心肌炎(28例)、扩张性心肌病(31例)和对照组19例。结果CAR阳性表达为棕色,主要表达于心肌细胞膜。在VMC心肌中均检测到较高水平的CAR表达(免疫组化面积积分为4.3975±0.0365);DCM组26例心肌组织中有较高水平的CAR表达(4.2064±0.0526),有5例几乎无表达(0.0723±0.0014),对照组心肌中几乎未检测到CAR的表达(0.0731±0.0362),VMC组及DCM组与对照组比较差异有显著性(P<0.05)。结论VMC和DCM心肌组织中CAR表达明显高于对照组,提示VMC和DCM的发病都有病毒感染的参与,两者可能来自同一个病因。     

病毒性心肌炎和扩张型心肌病心肌Fas蛋白表达

目的探讨病毒性心肌炎（viral myoearditis，VMC）和扩张型心肌病（dilated cardiomyopathy，DCM）的发病机制及相互关系。方法应用改良的免疫组织化学技术观察VMC、DCM和对照组（各20例）心肌Fas蛋白的表达，并对其阳性反应结果进行Ridit检验。结果与对照组相比，VMC和DCM组Fas蛋白阳性表达明显升高，经Ridit检验，3组阳性结果差异有统计学意义（P〈O．005）；进一步采用两两比较Ridit检验，对照组和其他两组之间差异有统计学意义（P〈O．05），而VMC和DCM组间差异无统计学意义（P〉0．05）。结论VMC和DCM心肌组织中Fas蛋白表达明显高于对照组，提示VMC和DCM的发病都有细胞凋亡的参与：凋亡可能是VMC引起心肌损伤及演变为心肌病的一个发病机制。     

病毒性心肌炎猝死心肌MCP-1表达及其法医学意义

目的探讨单核细胞趋化蛋白1（monocyte chemoattractant protein-1,MCP-1）在病毒性心肌炎（viral myocarditis,VMC）所致猝死者心肌组织中的表达及其意义。方法运用改良的免疫组织化学方法,观察VMC猝死组和对照组心肌组织中MCP-1的表达情况,应用统计学处理,比较两组间的表达差异。结果20例VMC猝死组中有17例MCP-1染色阳性;而20例非VMC猝死对照组仅4例局部散在MCP-1弱阳性表达,其余均阴性。VMC组MCP-1阳性表达率明显高于对照组（P〈0.01）。结论MCP-1的免疫组织化学检测可作为法医病理学诊断VMC所致猝死较为客观的病理形态学指标之一。     

Spry1和MAPK蛋白在病毒性心肌炎心肌组织中的表达

目的通过探讨Spry1和MAPK蛋白在病毒性心肌炎(viral myocarditis,VMC)心肌组织中的表达,揭示其发病及猝死机制,并为法医学心源性猝死的鉴定提供指导。方法 30只Balb/c雄性小鼠随机分为VMC组和对照组,分别腹腔注射柯萨奇B3病毒和Eagel’s培养液,处死小鼠取心脏,进行常规病理检查,并通过免疫组织化学、Western印迹法及real-time PCR方法检测心肌组织中Spry1蛋白及mRNA、MAPK蛋白表达变化。结果 VMC组在光镜下可见心肌间质水肿,广泛炎症细胞浸润,心肌细胞坏死,心肌纤维形成多处灶性、大片状坏死,心肌组织中Spry1蛋白表达水平低于对照组(P0.05),Spry1 mRNA水平略降低(P0.05);而MAPK蛋白表达水平在VMC组高于对照组(P0.05)。结论 Spry1参与的MAPK/ERK通路在病毒性心肌炎心肌纤维化过程中起着促进胶原表达的作用,从而导致心律失常、心功能减退甚至心源性猝死。     

DCM猝死与晚期DCM心肌Cx43表达的比较

目的观察和探讨扩张型心肌病(DCM)猝死与晚期DCM心肌中心肌连接蛋白(connexin,Cx)43表达差异及其意义。方法收集13例DCM猝死者心脏(A组)和5例晚期DCM患者行心脏移植手术切除的心脏(B组),运用免疫组化染色和图像分析技术,检测心肌Cx43阳性表达产物的平均光密度(OD)值和面积(S)值,比较两组间和左右心室间的差异。结果 Cx43阳性表达在A组明显减少,分布不均;在B组表达清晰,主要位于闰盘处。A组与B组心肌Cx43的S值之间的差异有统计学意义(P0.01),而各组左、右心室肌间相比,差异无统计学意义(P0.05);心肌Cx43的OD值,在A组与B组之间,以及各组左、右心室肌间差异均无统计学意义(P0.05)。结论 DCM猝死心肌Cx43表达较晚期DCM心肌明显减少,这种变化可能与DCM患者因心律失常发生猝死有关。     

病毒性心肌炎和扩张性心肌病中Dystrophin蛋白的表达

目的探讨病毒性心肌炎和扩张性心肌病的发病机制及相互关系,从而提高心性猝死法医学鉴定的可靠性和准确性。方法对17例对照(包括正常心脏、冠心病、高血压性心脏病等),25例病毒性心肌炎和28例扩张性心肌病的心肌组织进行改良的病理学dystrophin免疫组织化学研究。结果dystrophin蛋白在对照组,病毒性心肌炎组和扩张性心肌病组中阳性表达率分别为100%,88%,57%,三组表达差异有显著性(P<0.05),且在病毒性心肌炎和扩张性心肌病组间表达有显著差异(P<0.05),经Spearman等级相关分析呈显著负相关(r=-0.526)。结论病毒性心肌炎和扩张性心肌病心肌中细胞骨架蛋白均有破坏,且随着由病毒性心肌炎进展为扩张性心肌病,dystrophin蛋白表达逐渐降低,说明在病毒性心肌炎和扩张性心肌病的发病机制中可能与dystrophin的被破坏有关,病毒感染并破坏心肌细胞骨架蛋白并最终导致心肌细胞坏死,心功能受损,从而使病毒性心肌炎进展为扩张性心肌病。     

病毒性心肌炎和扩张性心肌病心肌中β-sarcoglycan的免疫组化染色观察

目的探讨病毒性心肌炎(viralmyocarditis,VCM)和扩张型心肌病(dilatedcardiomyopathy,DCM)的发病机制及相互关系,从而提高心性猝死法医学鉴定的可靠性和准确性。方法对17例对照组(包括冠心病、高血压性心脏病和正常心脏等),25例VCM和28例DCM的心肌组织进行改良的β-sarcoglycan免疫组织化学染色观察,并对其阳性反应率进行χ2检验及相关分析。结果β-SG蛋白在对照组,VCM组和DCM组中阳性反应率分别为100%,80%,46.4%。经χ2检验,3组阳性反应率差异有显著性意义(P<0.05);用χ2分割法分析,VCM和DCM组间差异有显著意义(P<0.05),且Spearman等级相关分析呈显著负相关(rs=-0.605)。结论病毒性心肌炎和扩张性心肌病病变与β-SG的被破坏有关;随着VCM病变程度的加重,其可能发展为DCM。     

心脏性猝死心肌组织N-Cadherin、Bax表达变化及评价

目的观察心脏性猝死者(SCD)心肌组织的神经性钙粘附蛋白(N-Cadherin)和Bax的表达变化,探讨其法医学意义。方法分别选取心脏性猝死和排除心脏疾病死因的尸检案例心肌组织标本各33例、29为SCD组和对照组。光镜下观察心肌组织病理学改变,检测N-Cadherin和Bax在心肌组织中的表达变化,并进行统计学分析。结果 N-Cadherin在SCD组心肌中表达呈弱阳性,排列紊乱,显著低于正常心肌,正常心肌组织中N-Cadherin呈强阳性表达,细胞间界限明显,排列整齐。Bax在SCD组表达呈阳性,显著高于正常心肌。结论 N-Cadherin和Bax的变化表达对心脏性猝死鉴定有意义。     

miR-17及其靶基因HIF-1α、STAT3在心源性猝死心肌组织中的表达及其意义

目的观察微小RNA17 (miR-17)与HIF-1α、STAT3在心脏性猝死(SCD)心肌组织中的表达水平,并探讨其意义。方法选择因心血管系统疾病猝死的20例心脏标本作为SCD组;选取20例死因为非心脏疾患的心脏标本作为对照组。应用免疫组织化学染色方法分别检测两组心肌组织中HIF-1α和STAT3表达,采用荧光定量PCR方法检测两组心肌组中miR-17、HIF-1α和STAT3的表达。结果免疫组化染色结果显示,两组心肌组织中HIF-1α、STAT3的表达比较,差异均有统计学意义(Z=-3.636、-3.552,P 0.01);两组心肌组织中HIF-1α与STAT3的表达水平呈负相关(r=-0.996,P 0.05)。荧光定量PCR方法检测结果显示,SCD组心肌组织中miR-17与HIF-1α相对表达量呈正相关(r=0.706, P 0.05),miR-17与STAT3相对表达量呈负相关(r=-0.730, P 0.05)。结论 SCD心肌组织中miR-17、HIF-1α高表达,STAT3低表达,miR-17、HIF-1α和STAT3联合应用有望作为诊断SCD较为客观的指标。     

心肌炎猝死者心、脾中柯萨奇 B3病毒基因的检测

目的检测病毒性心肌炎（ viral myocarditis,VMC）猝死者心、脾组织柯萨奇 B3病毒 (CVB3)基因,探索 VMC的病原学诊断方法。方法运用原位逆转录 PCR技术检测实验组（ VMC猝死, 8例）及对照组（非心性死亡, 4例）心、脾组织中的 CVB3基因。结果实验组中, 3例（第 1, 4, 7例）心肌 CVB3基因阳性, 4例（第 2, 4 , 6, 7例）脾组织 CVB3基因阳性;对照组心、脾 CVB3基因均为阴性。结论心、脾组织中 CVB3基因联合阳性可能是 VMC的重要特征;联合检测心、脾中 CVB3基因可提高 VMC的病原学诊断率。     

Presence of psychoactive substances in oral fluid from randomly selected drivers in Denmark

The aim of this investigation was to identify and characterise pathogenic mutations in a sudden cardiac death (SCD) cohort suspected of cardiomyopathy in persons aged 0-40 years. The study material for the genetic screening of cardiomyopathies consisted of 41 cases and was selected from the case database at the Institute of Forensic Medicine. Mutational screening by DNA sequencing was performed to detect mutations in DNA samples from deceased persons suspected of suffering from hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), and arrhythmogenic right ventricle cardiomyopathy (ARVC). A total of 9 of the examined 41 cases had a rare sequence variant in the MYBPC3, MYH7, LMNA, PKP2 or TMEM43 genes, of which 4 cases (9.8%) were presumed to be pathogenic mutations. The presumed pathogenic mutations were distributed with one case of suspected HCM and DCM (MYH7; p.R442H), one case of suspected DCM (LMNA; p.R471H), and two cases of suspected ARVC (PKP2; p.R79X and LMNA; p.R644C). The presented data adds important information on the genetic elements of SCD in the young, and calls for expert pathological evaluation and molecular autopsy in the post-mortem examination of SCD victims with structural anomalies of the heart.     

Myocardial Cx43 expression in the cases of sudden death due to dilated cardiomyopathy

PURPOSE: Probing into myocardial connexin (Cx) 43 expression in the cases of sudden death due to dilated cardiomyopathy (DCM) and relationship between Cx43 expression and sudden death. METHOD: Myocardial Cx43 was detected with immunohistochemical staining in the cases of 11 sudden death caused by DCM and 14 cases of control group who died of violent reasons and other diseases, which were autopsied in our department from 1997 to 2003. RESULT: Of 11 cases of DCM, there were 10 men and 1 woman with ranging in age from 7 to 49 years old (x (37.8) years old for 9 adult cases). Of 14 cases in the control group, there were 10 men and 4 women with ranging in age from 11 to 53 years old (x (29.9) years old for 11 adult cases). Myocardial Cx43 expression was obviously decreased in DCM group. Positive dyeing spots were different in size, distribution, color and disparity, some of them were distributed in the form of particle. Obvious change had not been observed in the cases of control group or with only slight changes in coloring degree and expressive area. The quantitative data showed that there was significant difference between two groups (p=0.0075) about Cx43 expressive area, but there was no difference between the left and right ventricles (p>0.05) in each group itself. And there was not difference between the two groups about average optical density of expression. CONCLUSION: Myocardial Cx43 expression is obviously reduced in the patients with DCM who die suddenly. The alteration of quantity and distribution of myocardial Cx43 expression is probably related to sudden death of the patients with DCM.     

Alcoholic cardiomyopathy versus chronic myocarditis--immunohistological investigations with LCA, CD3, CD68 and tenascin

Dilated cardiomyopathy (DCM) is a disorder of unknown aetiology characterized by the left ventricular cavity enlargement and wall thinning associated with reduced left ventricular wall motion. DCM in chronic alcoholics is supposed to be caused by alcohol induced myocardial damage (alcoholic cardiomyopathy). Nevertheless, cardiotropic viruses, such as enteroviruses have long been suspected as causative agents for at least some forms of DCM. In the present study, 13 cases of DCM in chronic alcoholics were investigated with qualification and quantification of infiltrating leucocytes using immunohistological antibodies against leucocyte common antigen (LCA), T-lymphocytes (CD3) and macrophages (CD68). In addition, the expression of tenascin, playing a role in the initiation of fibrotic changes, was examined. All antigens were known to be possibly enhanced in cases of chronic myocarditis. Using these immunohistological techniques, 2 out of 13 cases had evidence for chronic inflammatory myocardial alterations in the sense of lymphocytic infiltrates (>2.0 CD3 T-lymphocytes/visual field at 400 x (HPF); >7 CD3 T-lymphocytes per mm(2)). These cases were diagnosed as having inflammatory cardiomyopathy. The other cases without myocardial inflammation were diagnosed as idiopathic/alcoholic DCM.     

Fatal intoxication due to excessive dichloromethane inhalation

A 22-year-old white male was found dead at his working place in a car lacquering company. He had removed lacquer residues by using a solvent containing dicloromethane (DCM) without using a gas mask. Pathology revealed signs of asphyxiation with obvious petechial bleedings and expressed microthrombosis of the pulmonary arteries. Toxicological analysis showed excessive concentrations of DCM which are inhaled due to exposure of extreme air concentrations.     

Forensic medical estimation of the morphological changes in the myocardium and hypothalamo-pituitary-adrenal system in case of sudden cardiac death

This morphological, morphometric, and immunohistochemical study of the myocardium, adrenal gland, and hypothalamus included 50 corpses of men aged from 25-49 years deceased by reason of acute coronary failure associated with coronary heart disease. The commonest lesions in the myocardium were contractures (54.3 +/- 3.5% [51.2; 57.8%]), cyclic deformation (44.7 +/- 6.3% [38.2; 51.4%]), and dissociation of cardiomyocytes (61.2 +/- 2.3% [56.3; 64.8%]). The sinoatrial and atrioventricular nodes of the heart contained large amounts of adrenaline-positive cells (45.6 +/- 4.5% [40.1; 48.7%]) and 34.2 +/- 2.9% [31.2; 37.4%]). The equally large numbers of adrenaline-positive cells were detected in the adrenal medulla (67.2 +/- 6.8% [61.9; 74.3%]). The arcuate, supraoptic, and paraventricular nuclei of the hypothalamus underwent reversible changes of neutron content (the degree of damage 25.6 +/- 4.8% [21.2; 29.8%]) and contained large amounts of noradrenaline-positive and dopamine-positive neural cells (54.2 +/- 3.6% [51.4; 59.3%]) and 28.7 +/- 2.1% [23.4; 31.7%]) respectively. It is concluded that the morphometric and immunohistochemical changes detected in the present study can be used as the additional forensic medical criteria for diagnostics of death from acute coronary failure associated with coronary heart disease.