人体组织中ABH物质分布的研究

本研究采用特异性红细胞粘附试验(SRCA 试验)方法,系统地研究了11例已知 ABO 血型及分泌状态尸体的38种组织器官 ABH 物质的定位与分布,发现粘膜、粘液腺及前列腺中均含有丰富的 ABH 物质,并受分泌状态控制。血管内皮细胞、复层上皮细胞、胰腺腺泡上皮及汗腺中也含有较多的 ABH 物质,但不受分泌状态的影响。新发现肺泡上皮、肝小胆管粘膜上皮亦含有 ABH 物质。其它组织器官除自身的血管内皮及红细胞含有 ABH 物质外,均未测出 ABH 物质。采用 SRCA 试验,室温放置13天的皮肤组织亦能正确地测出其 ABO 血型。     

Determination of ABO antigens in fingernails using the APAAP (immunoalkaline phosphatase) technique

Fingernail specimens with adherent nail-bed were taken from autopsy material with blood groups A, AB, B and O. Frozen 4-5-microns sections were submerged and floated carefully during each working step. Portions of fingernails were contaminated with blood and buccal cells, respectively. Furthermore, fingernail fragments of 8 volunteers were embedded in a biocomponent adhesive according to Grieve and Kotowski (Forensic Sci. Soc., 26 29-34) (1986) and cut by the usual microtome technique. APAAP staining is a proper method for demonstrating blood group antigens in fingernails from groove to margin. Frozen sections as well as smallest specimen embedded in a suitable adhesive are applicable for staining procedures. Using freshly prepared artificial stains, blood group constellations of red blood cells and/or buccal cells adherent on the surface of fingernails may be distinguished from the nail matrix.     

抗原修复法和固定液pH值对切创皮肤免疫组化染色的影响

目的比较不同抗原修复方法对不同pH混合福尔马林固定液固定的切创皮肤的免疫组化染色效果。方法小鼠皮肤切创后3d取材,分别用pH为5.0、6.0、7.0、8.0、9.0的混合4%福尔马林固定液固定,经高压锅、微波、胰蛋白酶、胃蛋白酶、微波+胰蛋白酶法对抗原修复,观测caspase-3,caspase-6,caspase-7,IL-8,IL-10的免疫组化染色(SABC法)效果。结果热修复法阳性细胞率高于蛋白酶修复法,高压修复法阳性细胞率略高于微波修复;高压修复法的脱片现象高于微波修复,胃蛋白酶修复脱片高于胰蛋白酶修复;微波+胰蛋白酶双修复法阳性细胞率最高,不发生脱片现象;固定液pH值为7.0和8.0时,阳性细胞率最高,染色效果最好。结论pH7.0和8.0混合福尔马林固定液固定的切创皮肤,经微波+胰蛋白酶法修复抗原,免疫组化染色效果最佳。     

大鼠电击死心脑肺及皮肤超微结构的改变

目的 观察电击致死大鼠的心、脑、肺、皮肤超微结构形态学改变，寻找电击死的法医学鉴定依据。方法 大鼠12只，其中电击致死6只，另6只对照；取心、脑、肺、皮肤，用戊二醛固定、锇酸染色后透射电镜观察。结果 所取组织细胞均见明显细胞凋亡样改变，其中红细胞肿胀变形，充填整个毛细血管管腔。结论 心、脑、肺、皮肤组织细胞与血管内皮细胞的超微结构形态改变对鉴定电流死有参考价值。     

Immunohistochemical study of blood group activities in severely burned human tissue

The blood, livers and lungs obtained from donors or cadavers of known blood groups were experimentally burned, while the temperatures inside the tissue specimens were automatically measured. All the distinguishable portions with various thermo-changes in each burned tissue specimen were examined for their blood-group activities A, B, Lea, Leb and P1 by means of the immunofluorescence and immunoperoxidase techniques. In the layer immediately before charring of the blood masses (tissue temperature: ca. 200-250 degrees C), the activities A, B, Lea, Leb and P1 could be specifically demonstrated on the erythrocyte membranes. In case of the burned livers and lungs, the A- and B-activities could be detected in the small blood vessels in the layer immediately before charring (ca. 200-250 degrees C), though the layer was so severely thermo-changed, that their original tissue structures could be no more observed. In the severely thermo-coagulated, porous and hardened layer of both organ tissues, the A- and B-activities could be demonstrated on the endothelial cells of the hepatic sinus and small blood vessels (especially in the alveolar walls). The simply thermo-coagulated inner portions well retained their original tissue structures and the A- and B-activities remained on the epithelial cells of the alveoli and bronchioles of the lung as well as the cells described above. The Lewis blood-group activities could be demonstrated on the epithelial cells of the bronchioles in the simply thermo-coagulated inner portion of the lung. The P1-activity could not be demonstrated in the burned tissues of the livers and lungs.     

未知病变类型脑血管中β-amyloid、α-actin、collagen Ⅳ的含量

目的研究未知病变类型脑血管病变的结构特征。方法通过刚果红染色、免疫组织化学染色、计算机图像分析技术对未知病变类型脑血管病变的β-amyloid、α-actin、collagenⅣ的含量进行研究。结果未知病变类型脑血管壁α-actin、collagenⅣ呈少量阳性染色,与正常脑血管存在显著差异(P<0.05);β-amyloid染色呈阴性,与正常脑血管无差异(P>0.05)。病变血管壁中上述三种蛋白的表达特点与脑血管淀粉样变(cerebralamyloidangiopathy,CAA)及小动脉硬化玻璃样变不同。结论未知病变类型脑血管病变具有不同于CAA的病变特征。     

Determination of Both Fetus’ and Mother's Blood Type from an Autopsy Case Immersed in Formalin for Over 50 Years

A female fetus which had been immersed in formalin for more than 50 years was found in Japan. Because no liquid blood could be obtained, we tried to use immunohistochemistry (IHC) methods to tissue samples obtained at autopsy to identify both the fetal and mother's blood type. We detected B antigens on endothelial cells in paraffin sections of the fetal organs. Furthermore, we observed both anti‐A‐ and anti‐B‐positive red blood cells in the intervillous space, which is indicative of the mother's blood type. To our knowledge, this is the first case report on determining the blood type of both the fetus and the mother from tissue immersed in formalin for such a long time. The results suggest that IHC is valuable for the determination of ABO blood type in circumstances of long postmortem duration and unfavorable storage conditions.     

Intra-cartilaginous laryngeal haemorrhages and strangulation

A previously unrecognized laryngeal injury in young female victims of manual strangulation is described. Twelve larynges that were retrospectively and prospectively collected at the Office of the Chief Coroner for Ontario (1982–1997) were used for this study. In all instances, the larynges were from cases of strangulation (mean age 27±10 years, range 20–46) with classical postmortem findings of asphyxia and either manual or combined manual and ligature strangulation. None of the larynges had fractures of the lamina of the thyroid cartilage, superior cornua, or cricoid cartilages. However, in 9 of the 12 cases (75%), sagittal sections revealed multifocal acute haemorrhages into the base of the superior cornua of the thyroid cartilage at the point of origin from the laminae. The presence of acute intra-cartilaginous haemorrhage into the larynx likely represents the disruption of small blood vessels due to elastic deformation of the flexible larynx during strangulation. The recognition of this form of laryngeal injuries broadens the pathological findings in cases of asphyxia associated with pressure on the neck.     

Unusual skin lesions mimicking bruises caused by Pseudomonas septicemia secondary to undiagnosed peripheral T-cell lymphoma in a young woman

We present a case of a 30-year-old woman with learning difficulties who was found dead at home by her mother. Her body was partially naked and covered in a number of unusual skin lesions with a targetoid appearance with red erythematous centers and well-delineated halos of pallor. These lesions were initially thought to be bruises by the police and by a forensic postmortem instigated. Postmortem examination also identified hepatosplenomegaly, severe lymphadenopathy, and focal patchy colonic ulceration. Histologic examination of the skin and bowel ulcers showed the lesions to be areas of infarction caused by Pseudomonas aeruginosa vasculitis. Pseudomonas was also cultured from the swabs of the abdomen, the spleen, and the blood cultures. Histologic findings of the lymph nodes showed complete effacement of the normal architecture by a population of pleomorphic small lymphoid cells. Immunohistochemistry confirmed the predominant cell type to be T-cells. The diagnosis of peripheral T-cell lymphoma was made. The cause of death was given as Pseudomonas septicemia secondary to immunocompromise resulting from the undiagnosed peripheral T-cell lymphoma. The pathogenesis of Pseudomonas and its association with malignancy is discussed along with a brief review of peripheral T-cell lymphomas. This case report demonstrates the characteristic macroscopic appearance of cutaneous Pseudomonas-associated lesions and how they can be misinterpreted as bruises.     

Immunocytochemical identification of AB0 incompatible erythrocytes following fatal transfusion reaction

A patient with blood group O died 8 h after an accidental transfusion of one unit of A1 erythrocytes. The admixture of group A red cells was not detected during postmortem serology. Paraffin-embedded autopsy material was studied by the indirect immunoperoxidase technique using monoclonal antibodies. Group A red cells were easily identified as circulating agglutinates in the larger vessels, in the capillary vessels of all organs examined, and as closely packed cell aggregations in the sinuses of the spleen. However, there was no clear evidence of erythrophagocytosis in the spleen or in liver.     

Behavior of animal blood in blood typing systems. Isoelectric focusing of erythrocyte acid phosphatase and phosphoglucomutase

Isoenzyme band patterns of animal blood erythrocyte acid phosphatase (EAP) and phosphoglucomutase-1 (PGM) were studied by isoelectric focusing on ultrathin polyacrylamide gels. For blood from all animals tested (dog, cat, cow, sheep, and goat), the overall band patterns for both isoenzymes were different from those of the most common human types of these enzymes, although some animal EAP and PGM bands appeared in the human band areas. When mixtures of human and animal red blood cells were studied, it was found that misinterpretation of human types was possible only if the overall band pattern of the mixtures was ignored. For the animal blood tested, the strong PGM bands appearing outside the human band areas could be used as "markers" for the possible presence of animal blood in the samples tested.     

Blood vessels recovered with organs and intended for use in organ transplantation. Final rule

The Food and Drug Administration (FDA) and the Health Resources and Services Administration (HRSA) are amending their regulations to include as part of an organ those blood vessels recovered with the organ that are intended for use in organ transplantation (HRSA regulation); and to exclude such blood vessels from the definition of human cells, tissues, or cellular or tissue-based products (HCT/Ps) (FDA regulation). The purpose of this final rule is to amend the regulations so that blood vessels recovered with organs and intended for use in organ transplantation, and labeled as such, are governed by the regulations pertaining to organs. The regulation of other recovered blood vessels remains unchanged. We (HRSA and FDA) believe that this change will eliminate the burden resulting from an organ procurement organization's efforts to comply with both FDA and HRSA rules with respect to blood vessels (FDA jurisdiction) and organs (HRSA jurisdiction).     

A novel histological technique for distinguishing between epithelial cells in forensic casework

There are a number of forensic cases in which the identification of the epithelial cell type from which DNA originated would provide important probative evidence. This study aimed to develop a technique using histological staining of fixed cells to distinguish between skin, buccal and vaginal epithelium. First, 11 different stains were screened on formalin-fixed, wax-embedded cells from five women. Samples were analysed qualitatively by examining staining patterns (colour) and morphology (absence or presence of nuclei). Three of the staining methods – Dane's, Csaba's and Ayoub-Shklar – were successful in distinguishing skin epithelial cells from buccal and vaginal. Second, cells were smeared directly onto slides, fixed with one of five fixatives and stained with one of the three stains mentioned above. Methanol fixation, coupled with the Dane's staining method, specific to keratin, was the only technique that distinguished between all three cell types. Skin cells stained magenta, red and orange and lacked nuclei; buccal cells stained predominantly orange–pink with red nuclei; while vaginal cells stained bright orange with orange nuclei and a blue extracellular hue. This staining pattern in vaginal cells was consistent in samples collected from 50 women aged between 18 and 67. Identification of cell type from unlabelled micrographs by 10 trained observers showed a mean success rate of 95%. The results of this study demonstrate that histological staining may provide forensic scientists with a technique for distinguishing between skin, buccal and vaginal epithelial cells and thus would enable more conclusive analyses when investigating sexual assault cases.     

Blood vessels recovered with organs and intended for use in organ transplantation. Direct final rule

The Health Resources and Services Administration (HRSA) and the Food and Drug Administration (FDA) are amending their regulations to consider as part of an organ those blood vessels recovered with the organ that are intended for use in organ transplantation (HRSA regulation); and to exclude such blood vessels from the definition of human cells, tissues, and cellular and tissue-based products (HCT/Ps) (FDA regulation). We (HRSA and FDA) are taking this action to provide that blood vessels recovered with organs and intended for use in organ transplantation are governed by the regulations pertaining to organs. The regulation of other recovered blood vessels remains unchanged. We believe that this change will eliminate the unnecessary burden resulting from an organ procurement organization's efforts to comply with both FDA and HRSA rules with respect to blood vessels (FDA jurisdiction) and organs (HRSA jurisdiction). We are issuing these amendments directly as a final rule because they are noncontroversial, and there is little likelihood that we will receive any significant adverse comments. Elsewhere in this issue of the Federal Register, we are publishing a companion proposed rule under our usual procedures for notice and comment in the event that we receive any significant adverse comments on the direct final rule. If we receive any significant adverse comments that warrant terminating the direct final rule, we will consider such comments on the proposed rule in developing the final rule.     

Massive fat and necrotic bone marrow embolization in a previously undiagnosed patient with sickle cell disease

A case of sickle cell disease diagnosed postmortem is described. A 37-year-old black woman presented with anemia, respiratory distress, and abdominal and back pain. Death followed an intramuscular injection of iron, and anaphylaxis was clinically diagnosed. At autopsy, massive fat and necrotic bone marrow embolization of pulmonary and renal vessels was found. In the vertebral column, multifocal areas of ischemic necrosis were present, and proved to be the source of this embolization. Sickled red cells appeared in bone marrow sinusoids, and signs of disseminated intravascular coagulation were present.     

Immunohistochemical studies on postmortem lividity

An immunohistochemical investigation of postmortem lividity was performed to illuminate localization of hemoglobin (Hb) and the mechanism of fixed lividity. The fixed lividity was defined as an unfading phenomenon by thumb finger pressure. Skin specimens were taken from 68 autopsy cases 7.5–336 h (2 weeks) postmortem. Localization of Hb of the specimens was examined by a labeled streptabidin biotin (LSAB) method using polyclonal (rabbit antihuman hemoglobin antibody) and monoclonal (mouse anti-human hemoglobin monoclonal antibody) antibodies. Positive staining for Hb was observed in various sites of the skin, i.e. in only intravascular erythrocytes, in vascular walls and perivascular tissue including sweat glands and sebaceous glands, in the dermal connective tissue, and in almost all of skin tissue except the horny layer. The diffusion of Hb into skin tissue was observed in 20 of 41 displaceable lividity cases (49%) and 11 of 27 fixed lividity cases (41%). Compared to displaceable lividity, superficial plexi in fixed lividity were filled with erythrocytes, which were markedly immunodetected. These findings support the hypothesis that the fixation of lividity is not due to diffusion of Hb into skin tissue but hemoconcentration in blood vessels.     

电击伤最佳病理取材部位的初步研究

目的探讨日常低电压电击后机体电流通路上极向化等病变规律和最佳病理取材部位。方法SD大鼠随机分为电击组和对照组,建立左前爪-右后爪路径220V电击大鼠模型,取电极接触处皮肤、四肢血管、神经和周边组织及腹腔大血管,常规HE染色病理组织学图像分析。结果电极接触处皮肤基底细胞(skinbasalcell,SBC)核明显极向化改变,长/短径比值与对照组比较差异高度显著,P<0.001;电流通路上腕和踝部血管及腹主动脉内皮、平滑肌细胞核的极向化与对照组比较,差异显著,P<0.05,其中腕和踝部病理改变最明显;周围骨骼肌细胞核的长/短径比值无明显规律性。结论在电击死中最常见的“手-足通路”,腕、踝部血管可能为病理取材的最佳部位。同时,由于腕和踝部距离手脚掌较远,直接接触热源和化学试剂等其他可能引起“极向化”改变的因素机会少,因此,腕和踝部血管及其周边组织的极向化改变更具有电损伤的特异性。     

Difference between in vivo and postmortem distances between anterior chest and heart surface. A combined autopsy and in vivo computerized tomography study

The distance between anterior chest surface and intrapericardial surfaces of the heart and great blood vessels was measured on 37 cases (seven with acute fatal hemopericardium) at autopsy and on 24 live persons by computerized tomography. At autopsy, the apex of the heart was always closest to the skin surface except in cases with acute fatal hemopericardium, where the heart was displaced backwards by 10-40 mm. At computerized tomography, chest-heart distances were approximately 16 mm shorter than at autopsy. Changing the position of the patient from supine to prone decreased the distances by about 10 mm. The data presented demonstrate that the topography of the heart and great vessels is changing with the position of the body in vivo and that chest-heart distance tend to increase postmortem; therefore, the depth of a stab wound in the anterior surface of the heart as measured at autopsy should be regarded as a maximal estimate of the length of the stabbing weapon actually having penetrated the tissues.     

Evaluation of Skin Surface as an Alternative Source of Reference DNA Samples: A Pilot Study

An acceptable area for collecting DNA reference sample is a part of the forensic DNA analysis development. The aim of this study was to evaluate skin surface cells (SSC) as an alternate source of reference DNA sample. From each volunteer (n = 10), six samples from skin surface areas (forearm and fingertips) and two traditional samples (blood and buccal cells) were collected. Genomic DNA was extracted and quantified then genotyped using standard techniques. The highest DNA concentration of SSC samples was collected using the tape/forearm method of collection (2.1 ng/μL). Cotton swabs moistened with ethanol yielded higher quantities of DNA than swabs moistened with salicylic acid, and it gave the highest percentage of full STR profiles (97%). This study supports the use of SSC as a noninvasive sampling technique and as a extremely useful source of DNA reference samples among certain cultures where the use of buccal swabs can be considered socially unacceptable.