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 .     

Fn免疫组化染色在人病毒性心肌炎死后诊断中的应用

目的研究纤维连接蛋白(Fn)免疫组化染色对轻度病毒性心肌炎的病理学诊断价值。方法运用兔抗人Fn多克隆抗体对人类心肌炎心肌标本进行Fn-LSAB免疫组化研究。结果发现心肌炎心肌组织内Fn大量沉积,部分心肌细胞内Fn阳性。结论Fn-LSAB染色可显示轻度病毒性心肌炎的心肌损害,Fn沉积是心肌组织存在炎症性病理变化的可靠标志之一。     

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.     

实验性病毒性心肌炎纤维连接蛋白的免疫组化研究

探索轻度病毒性心肌炎的法医病理学诊断方法。以适量coxsackieB3病毒感染Balb/c小鼠 ,造成小鼠轻度病毒性心肌炎 ,对病鼠的心肌进行纤维连接蛋白的LSAB免疫组化染色。结果发现 ,心肌炎鼠心肌组织内Fn大量沉积 ,部分心肌细胞内Fn阳性。Fn -LSAB染色可显示病毒性心肌炎轻度的心肌损害 ,Fn沉积是心肌组织存在炎症性病理变化的可靠标志之一。     

Diagnosis of sudden unexplained death in epilepsy by immunohistochemical staining for prolactin in cerebral vessels

Sudden unexplained death in epilepsy occurs when epilepsy patients die suddenly and unexpectedly in the absence of recent tonic-clonic seizure activity. There is currently no known reliable indicator of acutely lethal seizure activity. Clinical studies record a relationship between recent (within 10-40 minutes) seizure activity and elevated serum prolactin levels, and postictal elevation of prolactin within peripheral vessels has proved clinically useful in determining recent seizure activity. The authors hypothesized that elevated prolactin could be detected in cerebral vessels by immunohistochemical stains, serving as a marker for sudden unexplained death in epilepsy. They conducted a retrospective study of individuals who died in their jurisdiction during the 14 years from 1986 through 1999. The study contained one group of individuals who died of sudden unexplained death in epilepsy, a group with epilepsy who died of some other cause, and a control group whose members died rapidly of a gunshot wound of the torso. Sections of hippocampus and neocortex were obtained and stained with a polyclonal prolactin antibody. No significant difference in the level of immunostaining for prolactin in cerebral vessels was found between the experimental and control groups. A review of the protocols used indicates that revision of certain aspects may provide better immunostaining and more conclusive results.     

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。     

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.     

Coxsackie B3 myocarditis in a case of sudden unexpected death in young athlete: histopathological, immunohistochemical and molecularpathological for diagnosis

We present two non fatal cases of intoxication with carbofuran (CBF) documented by hair analysis. Carbofuran and 3-hydroxycarbofuran (OHCBF, its main metabolite) hair concentrations were determined using a liquid chromatography-tandem mass spectrometry method. The obtained results were surprising if we consider several hair analyses previously published and based on a theory of the presence of xenobiotic in the only segment that comprised its intake. Among the two intoxication cases, we noticed the presence of CBF and OHCBF in hair segments corresponding to 45 days before, and more than 100 days after, the day of intoxication. Additionally, repeated hair samplings and subsequent analysis revealed a decrease of the carbofuran's concentration during the hair life.     

Determination of ABO blood grouping from human oral squamous epithelium by the highly sensitive immunohistochemical staining method EnVision+

Using the highly sensitive immunohistochemical staining method EnVision+, which employs a dextran polymer reagent for the secondary antibody, the detection of the ABH antigens was attempted in the oral squamous epithelium. This new technique uses monoclonal antibody as a primary antibody and it takes about three hours for staining. The time is much shorter than conventional absorption-elution testing or absorption-inhibition testing for the determination of ABO blood grouping. Secretor saliva samples were stained at strong intensity by the antibody, which corresponded to its blood group and anti-H. On the one hand, nonsecretor saliva samples were stained at strong intensity only by the antibody that corresponded to its blood group, and at weak intensity only by anti-H. Since human oral squamous epithelium antigens were stained specifically by this method, we can examine the ABO blood group of saliva samples and perform cytodiagnosis at the same time. Our research suggested that the EnVision+ Method is a useful technique for ABO blood grouping of saliva in forensic cases.     

人脑挫伤后S-100和GFAP的变化

应用免疫组织化学染色 ,观察人脑挫伤后 S- 10 0和 GFAP时间变化的规律 ,为脑挫伤形成时间的推断提供新的形态学依据。S- 10 0阳性细胞面积、灰度在伤后 48h显著增加 (P<0 .0 1) ;伤后 4d阳性细胞面积、灰度达到峰值 (P<0 .0 1) ;伤后 11d仍然保持较高水平 (P<0 .0 1)。GFAP阳性细胞面积、灰度伤后 2 4h显著增加 (P<0 .0 1) ;7d达高峰 (P<0 .0 1) ;以后有逐渐下降的趋势。伤后人脑组织 S- 10 0和 GFAP免疫组织化学染色阳性细胞面积、灰度改变有时间规律 ,两者可作为在 2～ 2 0 d内不同时间脑损伤的推断指标     

Delimitation of the time of death by immunohistochemical detection of thyroglobulin

To improve the possibilities to delimit the time of death after longer laytime it was examined if this is possible by immunohistochemical detection of thyroglobulin. The results show that in our examination material the colloid and the follicular cells of the thyroid glands of up to 5-day-old corpses produce a positive immunoreaction towards thyroglobulin in all cases whereas none of the corpses older than 13 days show such a reaction. This means that in case of a negative immunoreaction the time of death can be assumed to lie more than 6 days before the autopsy. The fact that a negative immunoreaction occurs consistently after 13 days leads to the conclusion that when thyroglobulin has been stained in a specimen, the death of the respective person must lie a maximum of 12 days earlier, whereby these time-limits may change in considerably different surrounding conditions.     

Delimitation of the time of death by immunohistochemical detection of thyroglobulin

To improve the possibilities to delimit the time of death after longer laytime it was examined if this is possible by immunohistochemical detection of thyroglobulin. The results show that in our examination material the colloid and the follicular cells of the thyroid glands of up to 5-day-old corpses produce a positive immunoreaction towards thyroglobulin in all cases whereas none of the corpses older than 13 days show such a reaction. This means that in case of a negative immunoreaction the time of death can be assumed to lie more than 6 days before the autopsy. The fact that a negative immunoreaction occurrs consistently after 13 days leads to the conclusion that when thyroglobulin has been stained in a specimen, the death of the respective person must lie a maximum of 12 days earlier, whereby these time-limits may change in considerably different surrounding conditions.     

Delimitation of the time of death by immunohistochemical detection of calcitonin.

To improve the possibilities of delimitating the time of death after longer laytime it was examined if this is possible by immunohistochemical detection of calcitonin. The results show that in our examination material the c-cells of the thyroid glands of up to 4-day-old corpses produce a positive immunoreaction towards calcitonin in all cases whereas none of the corpses older than 13 days show such a reaction. This means that in the case of a negative immunoreaction the time of death can be assumed to lie >4 days before the autopsy. The fact that a negative immunoreaction occurred consistently after 13 days leads to the conclusion that when calcitonin has been stained in a specimen, the death of the respective person must lie a maximum of 12 days earlier, whereby these time-limits may change in considerably different surrounding conditions.     

不同部位颅脑摔跌伤的特征分析68例

目的探讨不同受力部位颅脑摔跌伤后损伤程度和特征差异。方法收集摔跌所致颅脑损伤68例,根据案情、受伤过程、临床体征、CT影像资料将受伤部位分为颅前部、颅侧部、颅后部;损伤类型分为颅骨骨折、硬脑膜外血肿、硬脑膜下血肿和脑挫裂伤;损伤数量分为单一和多发伤;颅骨骨折分为受力点骨折和非受力点骨折;脑挫裂伤分为冲击伤、对冲伤等。运用非参数统计χ2检验比较其频数分布。结果不同摔跌部位颅骨骨折和颅脑损伤程度存在明显差异(P0.05),颅后部摔跌颅骨骨折远离受力点多见,多发伤为主,而颅前部、颅侧部受力点骨折多见,单处损伤多见;颅后部、颅侧部摔跌后脑挫裂伤发生率高,以对冲部位多见,颅前部摔跌后脑挫裂伤发生率低,冲击伤亦多见。结论摔跌部位不同颅脑损伤程度和特征存在差异,CT检查资料有助于损伤机制推断。     

The ABO blood grouping of a minute hair sample by the immunohistochemical technique

The unlabeled antibody (PAP) immunoperoxidase technique was applied to the ABO blood grouping of human scalp hairs. Hair samples were subjected to longitudinal- or cross-sectioning, thus obtaining suitable samples for subsequent immunostaining. The immunostaining was carried out using rabbit anti-A and anti-B sera as the primary antibodies. With this technique, the group-specific staining which is revealed as a dark brown precipitate was clearly observed within the medullae of the hair shaft, and depending on the presence or absence of these precipitates, respective blood groups of unknown hair samples were determined. At the hair root, on the other hand, positive stainings were observed not only in medullary cells but also in some cortical cells of the keratogenous zone. From the present study, it can be safely said that this technique is of practical use for the ABO blood grouping from a minute (less than 3 mm) hair sample.     

Delimitation of the time of death by immunohistochemical detection of insulin in pancreatic beta-cells

To improve the possibilities to delimitate the time of death after longer laytime, it was examined if this is possible by immunohistochemical insulin detection. The results show that in our examination material, the pancreatic beta-cells of up to 12-day-old corpses produce a positive immunoreaction towards insulin in all cases, whereas none of the corpses older than 30 days show such a reaction. This means that in case of a negative immunoreaction, the time of death can be assumed to lie more than 12 days before the autopsy. The fact that a negative immunoreaction occurs consistently after 30 days leads to the conclusion that when insulin has been stained in a specimen, the death of the respective person must lie a maximum of 29 days earlier, whereby these time-limits may change in considerably different surrounding conditions.     

Determination of fetal age by immunohistochemical estimation of surfactant-producing alveolar type II cells.

We monitored the immunohistochemically determined amount of surfactant-producing alveolar type II cells in human fetal lung using polyclonal antibodies against apoprotein B and C of human pulmonary surfactant. Lungs of 30 dead-born fetuses without lung affection aged between 15 and 38 weeks of gestation were evaluated and the surface density of surfactant-producing alveolar type II cells was determined by morphometry. In lungs of fetuses with a gestational age less than 22 weeks no relevant number of positively reacting cells could be found. Between the 22nd and 29th week a progressive increase with considerable inter-individual variability was observed. From the 30th week on the number of the type II pneumocytes appeared rather constant without further significant increase. We provide evidence that the immunohistochemical detection of surfactant-producing alveolar type II cells is useful for the determination of the age of unknown and especially fragmented fetuses: The lack of surfactant-producing alveolar type II cells in fetal lungs before the 22nd week allows a rather safe distinction between fetal lungs of higher age from those of lesser age. Between the 22nd and 29th week an age-dependent increase in the number of these cells occurs with wide inter-individual variability allowing only an approximate age determination. In particular, this may be an important piece of information in fragmented fetal corpses. Furthermore, the number of surfactant-producing alveolar type II cells provides additional information on pulmonary maturation and may thus be helpful in the estimation of a theoretical survival chance.     

Delimitation of the time of death by immunohistochemical detection of glucagon in pancreatic alpha-cells.

To improve the possibilities of delimitating the time of death after longer laytime it was examined if this is possible by immunohistochemical glucagon detection. The results show that in our examination material the pancreatic alpha-cells of up to 6-day-old corpses produce a positive immunoreaction towards glucagon in all cases whereas none of the corpses older than 14 days show such a reaction. This means that in the case of a negative immunoreaction the time of death can be assumed to lie more than 7 days before the autopsy. The fact that a negative immunoreaction occurs consistently after 14 days leads to the conclusion that when glucagon has been stained in a specimen, the death of the respective person must lie a maximum of 13 days earlier, whereby under markedly different conditions to the ones of the cases here examined, a negative immunoreaction could happen earlier and a positive immunoreaction even later.     

图像分析技术测定DNA含量变化推断死亡时间的研究及应用前景

图像分析技术是20世纪50年代发展起来的一门技术,它通过对图像信息的收集、处理、计算、分析而得出图像各部分的数量变化,在生物医学领域已经成为一种常规的定量技术。本文对图像分析技术测定DNA含量的原理、死后细胞核DNA降解的规律及图像分析技术定量DNA推断死亡时间的应用前景进行了综述。     

Diagnosis of drowning by investigation of left heart blood

Nineteen cases of drowning and fourteen controls were studied in order to find pulmonary alveolar macrophages, diatoms and various, mainly crystalline, dirt particles in left heart blood. The results show promise for the development of a new diagnostic method for forensic purposes.