A preliminary immunohistochemical study of fibronectin in viral myocarditis

For postmortem diagnosis of viral myocarditis, 12 specimens of heart in autopsy were studied immunohistochemically with anti-fibronectin(FN) antibody. Among 5 cases with definite or suspected myocarditis, intensive FN-positive could be found in all monocytes, macrophages and some neutrophils and there were 3 cases with FN-positive cardiomyocytes. While in 3 violent death cases with leucocytes infiltration in the interstitial tissue of myocardia, no FN-positive cardiomocytes were observed, which was the same as that of 4 case without cardiac pathological changes. Moreover, there were rarely FN-positive leucocytes in the 7 cases without viral myocarditis. The results suggest that the immunohistochemical observation of FN in myocardia might be of value for detecting slight degeneration of cardiomyocytes and determining the inflammatory leucocyte infiltration in myocarditis.     

Heroin associated nephropathy – a post-mortem study

Renal specimens were obtained from 179 autopsies of persons autopsied in the Institute of Forensic Medicine, University of Bonn, from 1987 to 1997. All persons were known as intravenous drug addicts. All renal specimens were examined with hematoxylin-eosin, PAS, Siriusred and Gomori (methenamine silver trichrome stain) and investigated with primary antibody against LCA (leucocyte common antigen), CD 68, IgG and IgM. 105 specimens (61.7%) showed a mono-lymphocytic membranoproliferative glomerulonephritis (MPGN), 48 specimens (45.7%) deposits of IgM. No case with focal segmental glomerulosclerosis (FSGS) as reported in male African–American intravenous drug addicts was found. In 37 out of 54 cases, hepatitis antibodies were detected in serum and three out of these 54 cases were HIV-positive. Chronic hepatitis B and C are known to be associated with glomerulonephritis. We found some cases without detection of hepatitis antibodies but with severe glomerulonephritis. In contrast to African–American drug addicts, European drug addicts do not develop a FSGS but a MPGN, partly due to heroin or other adulterants and apparently independent from hepatitis infection.     

病毒性心肌炎肌动蛋白的免疫组化观察

对１０例尸检心脏标本进行了肌动蛋白（Ａｃｔｉｎ,Ａｔ）的免疫组化观察。依据Ｄａｌｓ标准的明确或界限性病毒性心肌炎５例,心肌组织内Ａｔ含量减损,呈局灶性或弥漫性淡染,偶见小灶性单个心肌细胞内Ａｔ缺失。心肌梗死２例,心肌组织内Ａｔ大片缺失;非心性死亡对照组３例,心肌组织内Ａｔ呈均匀一致的强阳性染色。结果表明,心肌组织内Ａｔ的免疫组化观察可望提高对不典型病毒性心肌炎诊断的敏感性,同时对区分轻度缺血性与炎症性心肌细胞损伤有参考价值     

Hypersensitivity myocarditis and hepatitis associated with imipramine and its metabolite, desipramine

We present two cases of myocarditis and hepatitis with histologic characteristics of hypersensitivity-mediated drug reactions associated with imipramine and its metabolite, desipramine. In one case, death was directly attributed to myocarditis; in the second case, the patient died of an acute myocardial infarct, but myocarditis may have played a contributory role. One patient was taking imipramine, and therapeutic concentrations of imipramine and desipramine were documented in postmortem blood. The other patient was receiving desipramine documented by in-patient hospital medication records. Both cases had liver lesions associated in the medical literature with adverse drug reaction to imipramine. Although myocarditis has been previously associated with amitriptyline, these cases appear to be the first reported in association with imipramine/desipramine. The fact that one patient was taking only desipramine suggests that it may be the offending agent.     

Sudden death caused by myocardial tuberculosis: case report and review of the literature

A 25-year-old fit man died suddenly while playing social soccer. Autopsy revealed an infiltrative lesion involving the left ventricle with overlying pericarditis. No other significant pathologic changes were observed. Histologic examination showed necrotizing granulomatous inflammation. No acid-fast bacilli were demonstrated in the pericardial fluid or on histologic examination. The presence of Mycobacterium tuberculosis DNA complex was confirmed by use of the ligase chain reaction technique. The differential diagnosis of myocardial tuberculosis includes sarcoidosis, rheumatic fever, rheumatoid arthritis, giant-cell-containing tumors, idiopathic (giant-cell) myocarditis, and bacterial infections such as tularemia and brucellosis. This case illustrates the protean manifestations of tuberculosis and highlights the use of molecular biologic techniques in arriving at a definitive diagnosis in cases of suspected tuberculosis.     

178例心源性猝死法医组织病理学诊断分析

目的探讨心源性猝死(SCD)的病理特点与鉴别要点。方法对四川华西法医学鉴定中心2000—2005年尸检出的178例SCD死亡案例进行回顾性分析,主要对其病因、年龄、诱因及病理改变进行分析。结果本组资料显示冠心病、心传导系统病变、心肌炎、心肌病等在SCD中占有较大比例。冠心病猝死是中老年人SCD的最主要原因,青壮年人SCD的病因以非冠心病为主。如传导系统病变、心肌炎、心肌病。在儿童的SCD中先天性心脏病、传导系统病变及心肌炎占主导地位。结论不同的病因,其病理特点不同,其病理变化是法医学鉴定的主要依据。     

Pulmonary thromboembolism after air travel: Two case reports, the review of literature and forensic implications

Anabolic androgenic steroids (AAS) are the main class of doping agents and their consumption produces adverse effects involving several organs and systems. Three cases of sudden cardiac death (SCD) and one of death due to congestive heart failure of previously healthy athletes who were AAS users are herein reported. Concentric cardiac hypertrophy with focal fibrosis (one case), dilated cardiomyopathy with patchy myocyte death (two cases) and eosinophilic myocarditis (one case) were observed and most probably relate to the final event. Molecular investigation for viral genomes was positive in one case (Ebstein virus). Our data confirm previous findings, showing that the most typical cardiac abnormality in AAS abusers is left ventricular hypertrophy, associated with fibrosis and myocytolysis. An exceptional cardiovascular substrate was represented by the case with drug induced eosinophilic myocarditis. These features are at risk of ventricular arrhythmias as well as congestive heart failure. The cause-effect relationship between AAS abuse and cardiac death can be established only by a rigorous methodology with the use of standardized protocols, including precise morphological studies of all target organs to search for chronic toxic effects. Laboratory investigations should focus on AAS searching on a wide range of biological matrices to demonstrate type, magnitude and time of exposure.     

Right ventricular damage due to pulmonary embolism: examination of the number of infiltrating macrophages

To investigate the pathological changes in the heart induced by pulmonary embolism, 20 autopsy cases of pulmonary embolism and 10 control cases of acute death from traumatic injury were examined. Adding to the routine hematoxylin-eosin (HE) staining, immunostaining with CD68 pan-macrophage marker was performed on the specimens obtained from both right and left ventricular walls. The number of macrophages was counted semi-quantitatively in 100 random high-power fields (HPF). Although typical pathological findings of myocardial infarction was not observed in any of the cases, 16 of the 20 pulmonary embolism cases showed an increase in the number of macrophages, mainly in the right ventricular wall. Four cases showed massive macrophage infiltration in the entire right ventricular wall. It is speculated that ischemia due to pulmonary embolism may be connected to its pathogenesis.     

Influenza A virus infection complicated by fatal myocarditis

Influenza virus typically causes a febrile respiratory illness, but it can present with a variety of other clinical manifestations. We report a fatal case of myocarditis associated with influenza A infection. A previously healthy 11-year-old girl had malaise and fever for approximately 1 week before a sudden, witnessed fatal collapse at home. Autopsy revealed a pericardial effusion, a mixed lymphocytic and neutrophilic myocarditis, a mild lymphocytic interstitial pneumonia, focal bronchial/bronchiolar mucosal necrosis, and histologic changes consistent with asthma. Infection with influenza A (H3N2) was confirmed by virus isolation from a postmortem nasopharyngeal swab. Attempts to isolate virus from heart and lung tissue were unsuccessful. Immunohistochemical tests directed against influenza A antigens and in situ hybridization for influenza A genetic material demonstrated positive staining in bronchial epithelial cells, whereas heart sections were negative. Sudden death is a rare complication of influenza and may be caused by myocarditis. Forensic pathologists should be aware that postmortem nasopharyngeal swabs for viral culture and immunohistochemical or in situ hybridization procedures on lung tissue might be necessary to achieve a diagnosis. Because neither culturable virus nor influenza viral antigen could be identified in heart tissue, the pathogenesis of influenza myocarditis in this case is unlikely to be the result of direct infection of myocardium by the virus. The risk factors for developing myocarditis during an influenza infection are unknown.     

应用 CD68免疫组织化学方法诊断不典型心肌炎

目的为划清心肌炎与心肌对机体某种病理状态的反应界线 . 方法应用 CD68免疫组织化学染色方法 , 对 26例 (6例典型急性心肌炎 ,20例明确或可疑心肌炎 )心源性猝死者的心肌作染色、光镜检查 . 取 10例正常心肌对比研究 . 结果心肌炎者病变部出现大量 CD68免疫组织化学染色阳性的单核细胞 . ( >20个 /× 40视野 ), 形态多样、堆集 ( >15个 ). 结论 CD68免疫组织化学染色可用作诊断不典型心肌炎的指标之一 .     

Analytical method for the determination of strychnine in tissues by gas chromatography/mass spectrometry: two case reports

This paper describes an analytical method for strychnine determination in biological samples by gas chromatography/mass spectrometry and their application in the investigation of two cases involving strychnine ingestion: A fatal case and a clinical one. The strychnine is isolated from biological samples using a liquid-liquid extraction procedure. The clean-up procedure is performed using an acid solution. Papaverine is used as internal standard in the quantification of strychnine. In the analysed specimens, the limits of quantification were 0.1 microg/ml or 0.1 microg/g. The recovery rate ranged from 75.0% to 98.7% and the coefficients of variation ranged from 4.8% to 10.5%.     

Cytomegalovirus-induced pneumonia and myocarditis in three cases of suspected sudden infant death syndrome (SIDS): diagnosis by immunohistochemical techniques and molecularpathologic methods

Immunohistochemical and molecularpathologic techniques have improved the diagnosis of myocarditis as compared with conventional histologic staining methods done according to the Dallas criteria. Additionally, immunohistochemistry and in situ-hybridization are able to demonstrate viral infection, e.g. cytomegaloviruses in salivary glands and lungs, locations both known to be involved in cytomegalovirusinfection. However, in many cases of proved cytomegalovirusinfection the cause of death remains unclear. We report on three children younger than 1-year of age, who died suddenly without prodromal symptoms. Their deaths were attributed to SIDS (sudden infant death syndrome). In situ-hybridization, immunohistochemical (LCA, CD45R0, CD68, MHC-class-II-molecules, E-selectine) and molecularpathologic investigations (PCR), however, suggested that death was caused by a cytomegalovirus-induced pneumonia or myocarditis. In the future, these methods should be used for investigating cases with suspicion of SIDS.     

Isolated myocarditis as a cause of sudden death in the first year of life

A series of three cases of isolated myocarditis, presenting as sudden death in infancy, occurred over a period of 3 months. This prompted a review of the autopsy records of the Children's Hospital of Winnipeg. Over a period of 40 years, 24 cases of isolated myocarditis were traced from 3196 autopsies. Most (21 of 24) cases of isolated myocarditis occurred in infants less than 12 months of age. In 16 of the infants there were either no antecedent clinical signs (sudden deaths), or a short clinical history of less than 24 h duration. Heart weights, however, were greater than the 99th percentile of published normals in three infants and above the 95th percentile in a further 16 infants. Areas of hypertrophied fibres were seen even in infants with a short history. These latter findings suggest that a latent phase of myocarditis may exist. The responsible pathogens were identified very rarely, due to a lack of suspicion of the existence of myocarditis, and it is suggested that samples of myocardium should be submitted for virologic examination in all cases of sudden death in the first year of life.     

病毒性心肌炎猝死心肌MMP-9和TGF-β1表达与心肌纤维化的关系

目的研究病毒性心肌炎心肌MMP-9、TGF-β1表达与心肌纤维化的关系,探讨其在病毒性心肌炎猝死鉴定中的作用。方法筛选30例病毒性心肌炎作为实验组（18例为明确性心肌炎组,12例为界限性心肌炎组）,另选10例非病毒性心肌炎作为对照组。Masson染色观察病毒性心肌炎心肌胶原增生的情况;免疫组化法检测MMP-9、TGF-β1的蛋白表达和组织定位,并用图像分析系统及统计学方法进行定量分析。结果病毒性心肌炎组心肌胶原纤维较对照组明显增多;MMP-9、TGF-β1的表达水平明显增高,MMP-9、TGF-β1的表达与心肌胶原纤维增生呈正相关。结论MMP-9、TGF-β1可能在病毒性心肌炎心肌纤维化过程中起重要作用,可以作为病毒性心肌炎猝死的辅助诊断的指标。     

Expression of TGF-beta 1 and proliferation of collagen in myocardium in viral myocarditis

To investigate the tissue repair of acute viral myocarditis, and to explore the diagnostic method of slight viral myocarditis in forensic pathology. Slight viral myocarditis model was induced in Balb/c murine by CVB3. Collagen proliferation in myocardium of mice with myocarditis was observed by special staining. The hearts of mice and human(9 cases) with myocarditis were studied LSAB-immunohistochemically with anti-TGF-beta 1 antibody. In the study, the proliferation of collagen was seen in myocardium in acute viral myocarditis. Generous expression of TGF-beta 1 was found in the myocardium of mice and human with myocarditis. The quantity of collagen proliferation and expression of TGF-beta 1 was positive correlation. It is concluded that the tissue repair exists in acute viral myocarditis and that positive staining of myocardium for TGF-beta 1 is a sensitive index of myocardial damage and tissue repair.     

Immunohistochemical detection of fibronectin and tenascin in incised human skin injuries

Immunohistochemical detection of molecules involved in inflammatory reaction can be useful for the diagnosis of vitality in skin wounds. We studied the expression of fibronectin (FN) and tenascin (TN) in 58 human skin wounds (48 vital and 10 postmortem). The age of vital injuries ranged from 3 min to 8 h and postmortem specimens were collected after a postinfliction interval of 15-180 min. One hundred thirty-seven formalin-fixed paraffin-embedded sections (mean: 2.3 sections per case) were stained with each of two monoclonal antibodies against FN and TN using the streptABC technique. A reticular staining for FN in wound edge and dermis was observed in 50% of vital specimens versus 0% in postmortem cases. Immunoreactivity was reduced in 10 autolysed cases. FN positivity exclusively at the injury margin was observed in 39.4% of vital wounds and 10% of postmortem cases. TN was negative in all specimens. Vital and postmortem hemorrhage areas showed positivity for FN and TN. Due to its low sensitivity, immunohistochemical analysis of FN is useful for determining vitality only in a minority of cases. Different factors in everyday practice, including autolysis and technical problems often produce false negative reactions with the result that FN cannot be regarded as a reliable parameter of vitality. Positive reactions (network staining) are more valuable than negativity but are not pathognomonic. Both vital and postmortem hemorrhages show an enhanced positivity for FN and TN, thus impeding the diagnosis.     

Diagnosis of non-typical myocarditis by applying immunohistochemical method of CD68

OBJECTIVE: To make a distinction between myocarditis and the reaction to some pathological state of myocardium. METHODS: Myocardium of 26 cases with sudden cardiac death were stained and LM light microscopies with immunohistochemical method 10 cases with normal myocardium were contrasted. RESULTS: A great deal of stained positive monocyte of immunohistochemistry emerged in the parasetions of myocarditis patients with various farms and stacking(> 15). CONCLUSION: The stain of immunohistochemistry can be used as one of the indications for diagnosing non-typical myocarditis.     

Tissue-specific differences in mRNA quantification of glucose transporter 1 and vascular endothelial growth factor with special regard to death investigations of fatal injuries

Glucose transporter 1 (GLUT1) and vascular endothelial growth factor (VEGF) have been established as being responsible for cellular adaptation to oxygen deficiency in tissue ischemia and hypoxia mediated by hypoxia-inducible factor 1. We hypothesized that mRNA quantification of these factors in autopsy tissue specimens could have diagnostic significance for investigating the pathology of death, especially after injury. Various cases (total, n=119; less than 48h postmortem) were examined, including fatal blunt injury (n=71) and sharp instrument injury (n=18), as well as asphyxia (strangulation/hanging, n=12) and acute myocardial infarction/ischemia (n=18) as controls. Quantification of mRNA by TaqMan real-time RT-PCR and immunostaining were performed for GLUT1 and VEGF in lung, kidney, and skeletal muscle specimens. The postmortem interval showed no significant influence on the relative quantification of mRNA during the early postmortem period. Characteristic results were found in blunt injury cases: both GLUT1 and VEGF mRNAs decreased in the lung but increased in the skeletal muscle depending on survival time. In the kidney, subacute deaths showed higher GLUT1 mRNA levels compared with acute deaths from blunt injury, but no significant change was found for VEGF mRNA. Immunohistochemistry showed visually predominant GLUT1 immunoreactivity in the renal cortex for cases with a longer survival time, coincident with the results at the mRNA level. Tissue-specific differences in mRNA quantification of GLUT1 and VEGF shed light on tissue ischemia/hypoxia and subsequent tissue-dependent pathophysiological changes leading to death after injury.     

纤维连接蛋白在诊断心肌梗死的特异性研究

探讨纤维连接蛋白 (Fn)对心肌梗死死后诊断的特异性。应用免疫组织化学和图像分析技术 ,对正常心脏、心肌梗死及其它非梗死性因素 ,如心肌炎、窒息、电击死、出血性休克、心挫伤、有机磷中毒等 ,直接或间接引起心脏损害时心肌细胞内Fn的变化进行研究。结果发现 :Fn仅在心肌梗死与心肌炎病例出现阳性反应 ,其阳性反应面积同正常对照组存在显著性差异 ,在窒息、电击死、出血性休克、心挫伤、有机磷中毒等病例未见明显阳性反应。Fn作为心肌梗死死后诊断指标仅受心肌炎的影响 ,对诊断心肌梗死具有较好的特异性     

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.