冠心病猝死者心肌中SOCS-1和Bax的表达

目的采用免疫组化方法,观察细胞因子信号转导抑制因子-1(SOCS-1)和Bax在冠心病猝死(SCD)者心肌中的表达情况,探讨其对SCD诊断的意义。方法 25例诊断为SCD者心脏样本为实验组,25例非心血管疾病猝死者心脏样本为对照组。应用免疫组织化学方法检测SOCS-1和Bax在心肌中的表达,应用SPSS 17.0软件进行统计处理,组间比较采用秩和检验。结果 SCD猝死者心肌SOCS-1与Bax的表达明显高于对照组,Uc值SOCS-1为5.830 6,Bax为5.573,两种指标组间比较,差异均有显著性意义(P<0.01);SCD组中SOCS-1表达阳性以上且Bax表达弱阳性以上有22例,而正常对照组仅有1例。结论 SOCS-1与Bax均可能参与了细胞凋亡的过程,检测两种指标在心肌中阳性表达,可以为SCD的诊断提供客观依据,且联合检测可提高SCD诊断的特异性。     

冠状动脉粥样斑块中C-反应蛋白的免疫组化研究

目的探讨冠状动脉粥样硬化斑块中C-反应蛋白(CRP)对冠心病猝死(SCD)的诊断意义。方法从本教研室2001~2004年尸检案例中挑选68例案例资料和心脏标本,分为3组A组SCD(27例);B组冠心病非猝死者(21例);C组无明显动脉粥样硬化病变的死者(20例);应用免疫组化染色(SABC法)和图像分析技术,检测每例冠状动脉左前降支和右主支粥样硬化斑块内的CRP染色情况,并对所得数据进行统计分析。结果A组有20例CRP免疫组化染色强阳性,6例呈较强阳性,1例为弱阳性;B组中3例呈较弱阳性,11例微弱阳性,7例为阴性;C组均未见阳性反应。结论检测冠状动脉粥样硬化斑块中的CRP对SCD的死后诊断具有一定意义。     

VEGF免疫组化染色在冠心病猝死诊断中的应用

运用免疫组化SABC法和图像分析与统计学处理系统 ,对16例冠心病猝死和15例非冠心病猝死对照组尸检心脏标本心肌局部血管内皮生长因子 (VEGF)的表达进行了研究。结果 :16例冠心病猝心脏标本心肌梗死局部心肌均有VEGF的阳性表达 ,尤以心肌梗死灶边缘为甚 ,阳性表达率为100 %。15例非冠心病猝死对照组心脏标本仅有2例心肌局部散在有VEGF弱阳性表达 ,其余为阴性。将免疫组化染色结果进行图像定量分析与统计学处理 ,冠心病猝死组阳性指数为13.68±2.73 ,对照组为2.05±0.84 ,两组结果有极显著性差异 (P<0.01)。本研究结果提示 ,VEGF免疫组化染色可望为冠心病猝死的死后诊断提供一个比较客观的病理形态学依据     

冠心病猝死心肌纤维连接蛋白免疫组化染色观察

目的研究纤维连接蛋白（FN）免疫组化染色对冠心病猝死（SCD）的病理学诊断价值。方法用兔抗人FN多克隆抗体对人SCD心肌、颅脑损伤和病毒性心肌炎致死者心肌进行FN-SP免疫组化染色观察，用图像分析处理系统对FN免疫组织化学染色阳性反应产物面积定量，所得数据进行统计分析。结果SCD组16例心肌组织内FN大量沉积；颅脑损伤致死组心肌细胞内FN染色阴性，病毒性心肌炎致死组部分心肌细胞内FN阳性；3组心肌细胞内的阳性反应面积存在显著性差异（P〈0．05）。冠心病猝死组阳性反应面积（μm^2）为54143．28±8474．92；颅脑损伤致组阳性反应面积（μm^2）为527．99±105．04；病毒性心肌炎组阳性反应面积（μm^2）为5483．53±1219．91。结论冠心病猝死者心肌FN免疫组化检测可为死因诊断提供可靠依据。     

心肌病猝死者心肌连接蛋白43的免疫组化染色观察

目的探讨心肌病猝死者心肌连接蛋白43(Cx43)染色变化及其与猝死的关系。方法运用免疫组化和图像分析技术,分2组(A和B组)检测20例心肌病猝死者心室肌的Cx43染色情况;并与14例非心肌病猝死者(C组)的检测结果对照。结果扩张型心肌病(DCM)猝死组(A组,11例)心肌Cx43染色明显减弱,阳性着色斑点大小不等、深浅不一、分布不均,有的呈散在颗粒状;其它类型的心肌病猝死组(B组,9例)亦见类似变化;非心肌病猝死的对照组(C组,14例)未见明显变化。定量检测并经统计分析发现,Cx43蛋白染色阳性的面积,A组与B组和C组的差异有非常显著性意义(P<0.01),B组与C组的差异无显著性意义(P>0.05);而平均光密度各组之间的差异无显著性意义(P>0.05)。结论心肌病猝死者心肌Cx43免疫组化染色明显减弱,尤以扩张型心肌病明显;心肌病猝死者心肌Cx43变化可能与其猝死有一定关系。     

Cathepsin-L在缺血心肌中的表达

目的检测cathepsin-L(CTSL)在大鼠早期缺血心肌中的表达变化,并探讨其法医学意义。方法建立大鼠早期心肌缺血模型,设置早期缺血组(early ischemic myocardium,EIM)、非缺血组(non-ischemic myocardium,NIM)、假手术组以及空白对照组,采用real-time PCR方法检测大鼠缺血后15min,30min,1h和2h CTSL mRNA的表达量,并进行组内和组间比较。收集心源性猝死者缺血心肌和机械性损伤致死者正常心肌样本,采用免疫组化染色方法观察心肌组织中CTSL蛋白表达。结果 EIM组CTSL mRNA表达量与NIM组、假手术组及空白组相比,在15min时无统计学意义(P>0.05),在30min、1h和2h时分别升高1.4、3.1和4.5倍(P<0.05);其余3组间差异无统计学意义(P>0.05)。观察10例冠脉狭窄程度Ⅲ～Ⅳ级的心源性猝死(SCD)者缺血心肌,CTSL表达增强。结论大鼠心肌缺血后早期即可检测到CTSL mRNA的升高,并在SCD冠脉狭窄死者缺血心肌中高表达,提示CTSL可作为心肌缺血与SCD的参考指标。     

过敏性和冠心病猝死者心肌组织中肥大细胞类胰蛋白酶、脑利钠肽的表达

目的探讨肥大细胞类胰蛋白酶、脑利钠肽(brain natriuretic peptide,BNP)在过敏性猝死和冠心病猝死鉴别诊断中的意义。方法选取山西医科大学法医病理学教研室2010—2015年尸检案例心肌标本共30例,分为颅脑损伤致死组、过敏性猝死组、冠心病猝死组,每组各10例。采用免疫荧光染色和Western印迹法分析各组心肌组织肥大细胞类胰蛋白酶和BNP的表达。结果过敏性猝死组、冠心病猝死组心肌组织内肥大细胞类胰蛋白酶免疫荧光染色均出现阳性染色;三组间两两比较,表达差异均具有统计学意义(P0.05)。冠心病猝死组心肌组织内BNP的表达量高于过敏性猝死组、颅脑损伤致死组(P0.05),过敏性猝死组与颅脑损伤致死组之间差异无统计学意义(P0.05)。结论联合检测心肌组织内肥大细胞类胰蛋白酶、BNP有望为过敏性猝死和冠心病猝死的法医学鉴别诊断提供帮助。     

基质金属蛋白酶2在冠心病猝死心肌中的表达

目的探讨基质金属蛋白酶2（mmp2）在冠心病猝死（SCD）心肌细胞和间质中的表达与SCD的关系。方法选取本教研室2003年51例死亡鉴定病例,分为SCD组,患有冠心病但非SCD组（对照组1）,无严重冠脉粥样硬化（As）病变和其它心血管疾病组（对照组2）,无严重As病变但有其它心脏病组（对照组3）。采用免疫组化SABC法染色及图像分析技术,检测mmp2在各组心肌细胞和间质内表达的阳性率（R值）和平均灰度值（H值）,并比较各组间的差异。结果SCD组与3个对照组之间心肌细胞内romp2H值的差异均有显著性意义;SCD组与对照组2和3之间心肌间质内mmp2H值的差异有显著性意义;镜下各组心肌间质及心肌细胞内表达的阳性率差别明显。结论心肌间质及心肌细胞内mmp2同相表达增高与SCD的发生有密切关系,联合检测心肌和间质mmp2的表达对诊断SCD有重要意义。     

心肌及传导组织内3种蛋白的变化与SMDS的相关性

目的 观察心肌及传导组织内肌动蛋白、血浆白蛋白及纤维连接蛋白的染色变化，探讨青壮年猝死综合征(SMDS)死后诊断的新方法。方法 应用免疫组化S-P法，对SMDS及冠心病猝死者心肌及传导系统内肌动蛋白、血浆白蛋白和纤维连接蛋白的进行染色观察。结果 22例SMDS例猝死者心肌及传导系统内的肌动蛋白缺染17例；血浆白蛋白染色阳性18例；纤维连接蛋白染色阳性15例。并发现在SMDS中，8例有CCS严重病变。结论 心肌及传导系统内血浆蛋白、肌动蛋白改变与SMDS密切相关，部分SMDS病例死前存在早期心肌缺血或梗死的改变。     

Cytoskeleton immunohistochemical study of early ischemic myocardium

Previous studies on cytoskeletal changes of in vitro and in vivo animal models of ischemic myocardium have suggested the possibility of using alterations in cytoskeleton proteins as an early marker for the post-mortem diagnosis of myocardial ischemia in cases of sudden death due to coronary artery disease (CAD). In the present study, using the technique of ABC-immunohistochemistry, we examine the changes of three cytoskeletal proteins: vinculin, desmin and α-actinin in human myocardial samples taken from 14 cases of CAD sudden death and 13 cases of non-CAD death. Results of these examinations are compared with immunohistochemical changes of myoglobin and histochemical staining of hematoxylin and eosin and phosphotungstic acid, and Masson trichrome. Patchy and extensive loss of the three cytoskeletal proteins was demonstrated in the myocardium of victims who died 1 h or later following the onset of symptoms of ischemic myocardium. The pattern of cytoskeleton change is equivocal in the cases of CAD who died less than 1 h after the onset of symptoms and of the cases of non-CAD. In these cases, no significant histological change was observed. With less non-specific background changes and stronger positive staining, immunohistochemical staining of the three cytoskeletal proteins is more reliable than myoglobin, which has attracted the attention of many pathologists searching for anatomic evidence of ischemic myocardium in coronary artery disease.     

97例心性猝死的法医病理学分析

目的探讨心性猝死（SCD）的特点、病理基础及致死因素和诱因等。方法对本系2002年12月至2006年12月期间，所作450例法医病理检案的97例心性猝死案例进行分析研究。结果97例SCD患者中，冠心痛猝死38例，心肌炎23例，心肌痛16例，高血压性心脏病12例，主动脉瘤破裂4例，肺栓塞4例。结论SCD病程短骤、凶险，以老年男性多见，冠心病占首位。由于猝死的因素繁多，因此对猝死事件的法医学鉴定要根据其发生特征和变化规律，作出客观、全面、准确的签定结论。     

Differential accumulation of pulmonary and cardiac mast cell-subsets and eosinophils between fatal anaphylaxis and asthma death: a postmortem comparative study

The distribution profile of infiltrated mast cell-subpopulations and eosinophils in the lung and heart sections of the patients who died of severe allergic hyperresponsiveness, was investigated. Four study groups were designed comprising 9 cases who died in systemic anaphylaxis (Group I), 10 asthmatic individuals whose death were assigned to acute and severe bronchial asthma (Group II), 10 asthmatic cases who died from non-immunological diseases (Group III). Twenty consecutive autopsies of non-allergic subjects who died of unnatural causes (Group IV) served as control group in this study. Utilizing antibodies against human tryptase and chymase and a double immunohistochemical staining method, we distinguished successfully all three subsets of mast cells (MC), MC-TC (containing both tryptase and chymase), MC-T (containing only tryptase) and MC-C (containing only chymase) types, subdivided on the basis of the protease compositions of their secretory granules. In order to immunostaining eosinophils, we used antibody to major basic protein as a marker. We also measured postmortem blood tryptase, specific and total serum IgE. The intriguing finding of this study was the marked differences of cellular composition in the lung between fatal anaphylaxis and asthma death. Significant augmentation of MCs infiltrated in lung and heart sections of anaphylaxis patients and drastic infiltration of bronchial eosinophils in asthmatic death and consequent release of their related inflammatory mediators might explain the differential expression of the associated symptoms in these two groups. The anaphylactic deaths did show neither emphysema nor significant mucous bronchial secretions whereas all asthmatic deaths did. The degree of pulmonary congestion and edema was also more severe in anaphylaxis. This corresponded with the histological findings and the location and number of mast cell-subsets and eosinophils in the different compartments of the lungs. We have demonstrated that the third type of mast cell MC-C is only found in the lungs in anaphylactic deaths. The practical consequence of our study will be that it is now possible to confirm a suspicion of anaphylaxis death not only by measurements of serum mast cell tryptase, but also by immunohistochemical methods.     

Sudden cardiac death and myocardial ischemia indicators: a comparative study of four immunohistochemical markers

The postmortem diagnosis of acute myocardial infarction represents a current challenge for forensic pathologists, particularly when death occurs within minutes to a few hours after the ischemic insult. Among the adult population the single most important cause of sudden cardiac death (SCD) is the well-known atherosclerotic coronary artery disease, commonly asymptomatic or unrecognized. The recognition of early myocardial damage using routine hematoxylin and eosin (H&E) staining is possible only if death has occurred at least 6 hours after the onset of the ischemic injury. The usefulness of immunohistochemical markers to the diagnosis of early myocardial damage has been recently suggested because most of them can be visible even serologically as early as few minutes after the beginning of the symptoms. To evaluate the usefulness of plasma and cellular antigens, their distribution patterns have been studied among a group of 18 SCD cases in which a myocardial ischemia was strongly suspected. For the present study, 4 markers have been selected on the basis of their different diagnostic potential as follows: among the plasma markers the C5b-9 and fibronectin, among the cellular markers the myoglobin and cardiac troponin. The results show that only the study of multiple markers such as those selected can provide enough evidence of myocardial ischemia and/or necrosis, supporting the final diagnosis of SCD. No single immunohistochemical staining is ideal for diagnosing early myocardial ischemia but a set of markers can improve the ability of forensic pathologists to detect ischemic areas when no macroscopic or microscopic evidence of necrosis is available. However, the interpretation of data obtained in each individual cannot be isolated from the overall assessment of the factors (cardiopulmonary resuscitation and/or agonal artifacts) that can affect the expression of each marker.