Light-microscopic examination of ABH and Lewis antigens in human tracheal and epiglottic glands using the avidin-biotin-peroxidase complex technique

The localization of ABH and Lewis antigens was examined in formalin-fixed, paraffin-embedded human tracheal and epiglottic glands using monoclonal anti A, B, H, Lea and Leb antibodies. The mucous cells of the glands showed reactivity with antibodies corresponding to the respective ABO blood groups of the tissue donors. The mucous cells from one blood group A, Le(a-b-) individual showed no reactivity with any antibodies and those from another blood group A, Le(a-b-) individual showed reactivity only with anti A antibody. In individuals from blood group Le(a + b-) of all ABO groups, the mucous cells reacted exclusively with anti Lea. In blood group O, Le(a-b+) individuals, the mucous cells showed intense reaction with anti H and Leb antibodies and weak to moderate reactivity with anti Lea. In Le(a-b+) individuals of A1, B and A1B blood groups, the mucous cells showed strong reactivity with anti A and/or B antibodies, moderate with anti Leb, weak or no activity with anti Lea and absent with anti H. In blood group A2 Le(a-b+) individuals, the mucous cells stained with anti A were weakly stained or completely unstained with anti H antibody, but cells negative with anti A gave strong positive reactions with anti H antibody.     

唾液中Lewis血型物质的定量研究

应用时间决定性荧光免疫测定法（TR－FIA法），对129例健康成人唾液中Lweis及H1血型物质进行定量检测。Le阳性个体均不同程度地检出了Lea和Leb物质。Lea物质含量：Le（a＋b-）型＞Le（a－b＋）型；Leb物质含量：Le（a－b＋）型＞Le（a＋b－）型。Le（a＋b－）型的Lea物质＞Leb，Le（a－b＋）型的Leb物质＞Lea物质。Le阴性的部分个体未能检出Lewis物质，其余个体也仅检出微量。根据Lea和Leb物质在唾液中的相对含量，可以推测红细胞Lewis型，并提示Leb物质可能由Lea物质转化而形成。     

A capillary tube method for the Lewis typing of red blood cells

A simple and inexpensive capillary tube procedure, which can be applied in the forensic science laboratory, is described for the detection of the Lewisa (Lea) and Lewisb (Leb) antigens on red blood cells. This procedure will permit approximately 4000 tests to be performed from a single 2-mL bottle of Lewis antiserum.     

ABH and Lewis antigens in the tracheal glands. I. Lewis-positive individuals

Antigens A, B, H, Lea, and Leb were demonstrated in the tracheal glands of 15 Lewis-positive secretors and 15 nonsecretors by the indirect immunoperoxidase technique. The detection of group-specific ABH antigens in mucous epithelium and intraductal secretory fluid was dependent on the secretor character. Whereas determination of secretor character was sometimes unreliable with anti-A and anti-B, the findings obtained by additional labeling with UEA1 were consistently correct. The secretors showed minimal gland labeling with anti-Lea and intensive labeling with anti-Leb; the nonsecretors, intensive Lea labeling and weaker or absent Leb labeling. Consequently, the determination of secretor character by ABH labeling could be verified by the behavior of the Lewis antigens. Since both morphologic structures and epithelial antigens are highly resistant to putrefaction, ABO and secretor character can also be diagnosed in badly decomposed tracheal wall specimens.     

Studies and observations on Lewis grouping of body fluids and stains

Leb positive individuals may phenotypically express both Lea and Leb in their secreted body fluids. Therefore, the interpretation of a Le(a + ,b-), non-secretor result is dependent on the absence of Leb. This study emphasises the importance of accurate procedure and biased selection of antisera such that Leb is preferentially detected in comparison with Lea. The relationship of the ABO group to the expression of Le is discussed in conjunction with the selection of samples for testing antisera and inclusion as control standards.     

ABH and Lewis antigens of tracheal glands. II. Lewis negative individuals

Immunocytochemical studies were performed on tracheal wall samples embedded in paraffin; the samples were taken at 23 autopsies. In all cases, the red cells had been typed in postmortem serological studies as being Le(a-b-). Blood-group antigens were demonstrated by the indirect immunoperoxidase technique, using monoclonal Anti-A, Anti-B, Anti-Lea and Anti-Leb; H was detected by UEA 1. The secretor characteristics could clearly be diagnosed from the ABH staining pattern of the mucous glands. In 11 cases, the lewis antigen labeling patterns were identical to the group of Lewis-positive individuals. It seems probable, from the statistical point of view, that these 11 individuals were, in fact, Lewis-positive and that the negative serology resulted from deterioration of the cadaver blood samples. The immunocytochemistry was quite different in the remaining 12 cases: (a) secretors (n = 9) were completely negative for Lea, Leb was equally negative in one case, but in the remainder it was detectable within mucous epithelia in minimal amounts and in an atypical granular distribution; (b) nonsecretors (n = 3) reversely exhibited complete negativity for Leb but a minimal staining for Lea. These findings are in harmony with the well established Lewis serology typing of secretions in Lewis negative individuals. Thus, a minimal Lewis antigen biosynthesis and secretion seem to occur in the absence of the Le gene: A alpha-4-L-fucosyltransferase of low activity might be the product of the allele le.     

Lewis blood group determination in bloodstains by planimetric measurement of eluted monoclonal antibodies

Planimetric measurements were employed for reading the results of an elution test to determine Lewis blood groups in dry human bloodstains. In the absorption-elution test, two varieties of indicators were used to detect eluted Lewis antibodies. First, 64 blood-stains aged between 2 to 8 months were tested with glutaraldehyde (GLA)-treated erythrocytes (planimetric hemagglutination assay, PMHA). This method demonstrated that dry stains weighing approximately 0.4 mg (equivalent to 3 microliters of whole blood) were sufficient for detection of Lea or Leb antigen. Results were obtained within 1 h. Then, 37 of these stains were tested with Lewis substance-coated latex particles (planimetric latex agglutination assay, PMLA). The presence of Lea and Leb antigen were detected from dry stains weighing 0.1 mg (equivalent to 1 microL of whole blood) within 3 h. Both these assays are faster and simpler with accuracy than the enzyme-linked immunosorbent assay (ELISA). Latex particles coated with Lewis substance are, in particular, strongly agglutinated and show agglutination patterns more clearly than erythrocytes. The blind tests using these two methods properly classified 7 Le(a + b-) and 23 Le(a-b + ) bloodstains; whereas, 5 Le(a-b-) stains were undetermined by the criteria for these tests. These results indicate the usefulness of the PMHA and PMLA for typing Lewis blood groups from small bloodstains.     

Detection of Lea substance in saliva stains by enzyme-linked immunosorbent assay (ELISA) using anti-gum arabic serum

It is known that rabbit anti-gum arabic (GA) serum has cross-reactivity with Lea antigen, and that, by using this cross-reactive anti-Lea antibody, the presence of Lea antigen in red blood cells and saliva can be demonstrated with accuracy. We have devised a rapid and highly sensitive method for detecting Lea substance in human saliva by the enzyme-linked immunosorbent assay (ELISA) method using an anti-Lea antibody isolated from anti-GA serum by affinity chromatography on Synsorb Lea. The ELISA plate, coated with the specific anti-Lea antibody, adsorbed the Lea substance in saliva which was subsequently identified by adding enzyme labeled anti-Lea IgG in that order. The method could detect the Lea substance in Le(a+) saliva stains as small as 0.1 by 0.1 cm in size that had been stored at room temperature for three weeks and in Le(a+) saliva stains 0.7 by 0.7 cm in size that had been stored for ten years. This method seems to be useful for quantitative analyses of the Lea substance in various body fluids.     

非分泌型的基因型与血型物质的分泌研究

阐明非分泌型的基因型与血型物质分泌量和Lewis表型的关系。应用时间决定性荧光免疫测定法（TR－FIA），检测传统的A型Lewis阳性非分泌型个体唾液中H、A及Lewis抗原含量，并以序列特异性PCR，确定其基因型。非分泌型个体唾液中检出了高含量的Lea抗原，其中8例不同程度的检出了少量H、A和Leb抗原，其FUTZ位点为se2／se2纯合型，属Le（a＋b＋）型；1例基因型为se3／se5杂合型，未能检出H、A及Leb抗原，属Le（a＋b－）型。se2是弱分泌基因，se3及se5是非分泌基因。     

Forensic Imaging‐Guided Recovery of Nuclear DNA from the Spinal Cord*,†

Abstract: Our objective is to document the recovery of DNA from the spinal cord or surrounding dura mater in 11 cases of severely burned human remains. Radiographs established that portions of charred tissue contained spine segments. Multidetector computed tomography (MDCT) revealed that each spine specimen contained an intact spinal cord remnant. A full DNA profile was obtained from seven specimens using spinal cord dura mater in six specimens and spinal cord medulla in one specimen. A partial profile was obtained from four specimens (spinal cord dura mater, 2; spinal cord medulla, 2). Bone and muscle surrounding the spinal cord appear to insulate nucleic acid containing tissue from critical thermal degradation. The spinal cord, which is easily identified by MDCT examination of remains and easily recovered at the postmortem examination, can be a source of DNA with extraction yields comparable with other tissue sources. Specimens of dura mater are preferable as processing time is faster than bone.     

用ABC法鉴定眼镜蛇中毒死亡的实验研究

S-D 大白鼠皮下分别注射不同剂量的眼镜蛇粗毒,于死后不同时间取注射部位肌肉及肝、肾、心、肺等做冰冻切片和石蜡切片。用ABC 免疫组化法检测蛇毒,观察蛇毒的组织定位。在注射部位的骨骼肌内检出特异性染色的蛇毒。HE 染色各器官未见特征性改变。蛇毒量为2LD_(50) ,死亡24小时后取材仍有阳性反应。蛇毒多结合于注射部位的骨骼肌细胞膜上。注射11LD_(50) 的蛇毒后,发现肾小球毛细血管内皮细胞和肝细胞核上结合有蛇毒。家兔注射眼镜蛇毒或被眼镜蛇咬伤后,注射部位和咬伤部位肌肉中也可检出蛇毒。本实验首次使用ABC 免疫组化染色,取得了迄今为止直接在动物组织上对蛇毒定位检测的最好结果。     

Fatal outcome of Ecstasy overdose

Consumption of amphetamine derivatives has considerably increased in Germany since the early nineties. Again and again intoxications with lethal outcome have also been reported, especially after physical activities such as intensive dancing. The authors present a case of an obviously suicidal intoxication of a 21-year-old man who was found dead with marked cuts on the right forearm. Toxicological tests showed in particular 3, 4-methylene-dioxymethamphetamine (MDMA). The results of the hair analysis revealed chronic consumption, but no cellular liver damage could be demonstrated. When examining the body fluids and organs, the highest concentrations by far were measured in the lungs (36.6 mg/kg), the liver (29.7 mg/kg) and the brain (29.1 mg/kg). The concentration in heart blood amounted to 10.8 mg/kg and was thus markedly higher than in peripheral blood (7.2 mg/kg). In the muscles concentrations ranged between 14.3 mg/kg and 20.2 mg/kg. On the basis of these concentrations and the available pharmacokinetic data the amount of MDMA probably consumed is assessed. It is demonstrated that for this assessment the concentrations in the muscular system are of special importance, as redistribution of highly lipophilic substances from the surrounding tissue is possible also in peripheral blood.     

Immunochemical and chromatographic determination of ABH antigens in compact bone tissue

The identification of ABH antigens from compact bone tissue is known from many sources. The purpose of this study was to make a contribution to the localization of blood-group-active substances in compact bone tissue. A variety of preparation and identification methods were successfully used and compared. Samples were extracted from compact bone tissue, separated by HPTLC, and examined using the absorption-elution and PAP techniques. Additionally, the PAP technique was carried out on cryostat sections. Serologically active blood-group substances were consistently demonstrated in the organic components of the haversian canals.     

冬季且被严重焚烧的尸体上也会有昆虫

一例凶杀案件发生于广东省最冷的1-2月,尸体经过了焚烧至大部分组织炭化,但在该被害者身上仍旧发现了蝇类昆虫,应用这些昆虫的发育进度准确的推断出了死亡时间。与其它季节未经过焚烧的尸体比较,该现场昆虫的种类、数量均明显减少,昆虫也主要分布于尸体下方避风处的腐败物质中。另外,在该尸体上片状分布着一层霉菌,这些霉菌的状态显然有时间相关性,提示我们通过对霉菌的研究也能找到死亡时间的线索。     

过敏性休克死亡豚鼠器官中IgE的表达及其法医学意义

目的寻找法医学鉴定过敏性休克死亡的病理形态学诊断指标。方法利用组织芯片技术采用免疫组化SP法检测豚鼠过敏性休克死亡后0,6,12,24h等4个时间点的心、肝、肺、肾、脾、胃、肠、气管及扁桃体组织中的IgE的表达。结果实验组肺及气管组织IgE呈阳性表达,脾组织IgE呈弱阳性表达,以死亡即刻表达最强,且随死亡时间的延长而减弱,各时间点的显色信号强度存在显著性差异(P<0.05);实验组其它器官及对照组9个器官均无表达。结论应用免疫组化方法检测IgE在肺、气管、脾组织中的表达,可作为法医学鉴定过敏性休克死亡的病理形态学诊断指标;肺及气管组织IgE阳性强度随时间的延长而减弱,提示法医学鉴定过敏性休克死亡应尽早尸检。     

SEM/EDAX检测内脏异物元素成分诊断溺死

为探讨溺死诊断的依据和推断入水地点，用扫描电镜／X－射线谱仪（SEM／EDAX）检测19例溺死尸体及28只溺死兔的肺、肾、心、肝等组织异物颗粒及其元素成分和含量，并以陆地上死亡尸体14具及8只陆地上勒死兔（2只勒死后入水浸泡6天）作实验对照。结果发现，19例溺死尸体及28只溺死兔的肺边缘区呼吸性细支气管、肺泡管、肺泡囊及肺泡内均可见高能谱值的异物颗粒，其大小自数微米至数十微米不等，多为无定形异物颗粒或细小异物颗粒集落。其中元素成分为硅、铝、钙、铁、铬、钛、钼、铅、锡、铜、溴等，与入水地点溺液中所含元素成分相同；肾、心、肝组织异物颗粒检出率分别为77％、53％及47％，数量较少，颗粒较小，大小自1微米以下至十数微米。非溺死尸体及实验兔的肺、肾、心、肝组织未检出或偶尔检出异物颗粒，其元素成分多为铁、钙或硅等，可能是病理及生理性异物颗粒，如含铁血黄素、钙化灶等。用SEM／EDAX检测水中尸体组织中异物颗粒及其元素成分和含量，可以诊断溺死，推断入水区域，为明确案件性质和确定侦察范围提供科学依据。     

Microbiota Composition and Pulmonary Surfactant Protein Expression as Markers of Death by Drowning

Pathological diagnosis of drowning remains a challenge for forensic science, because of a lack of pathognomonic findings. We analyzed microbiota and surfactant protein in the lungs for a novel diagnosis of drowning. All rats were divided into drowning, postmortem submersion, and control groups. The water, lungs, closed organs (kidney and liver), and cardiac blood in rats were assayed by targeting 16S ribosomal RNA of Miseq sequencing. Lung samples were analyzed by immunohistochemical staining for surfactant protein A. The closed organs and cardiac blood of drowned group have a lot of aquatic microbes, which have not been detected in postmortem submersion group. Furthermore, intra‐alveolar granular staining of surfactant protein A (SP‐A) was severely observed in the drowned group than the postmortem submersion and control groups. The findings suggested that the presence of aquatic microbiota in the closed organs and increased expression of SP‐A could be markers for a diagnosis of drowning.     

皮肤烧伤愈合过程中磷酸化JNK的变化

目的观察烧伤后人皮肤磷酸化JNK（p-JNK）的变化特点,初步探讨烧伤后创面愈合的分子机制。方法取12例烧伤后住院植皮患者的烧伤周边区皮肤为烧伤周边皮肤组;另取正常皮肤12例为正常皮肤组。应用免疫组织化学方法和常规HE染色检测p-JNK在皮肤烧伤周边区和正常皮肤组织中的变化情况。结果正常皮肤组织中表皮基底层细胞的胞浆和胞核内有p-JNK阳性着色,阳性细胞率为（8.8±1.3）%。在烧伤周边皮肤组中,p-JNK阳性着色主要定位于表皮细胞和部分炎症细胞中,阳性细胞率上升至（31.2±3.3）%,明显高于正常皮肤组织（P〈0.01）。结论烧伤后人皮肤p-JNK的变化可能与创面愈合有关。     

豚鼠过敏性休克肺组织中类胰蛋白酶和胃促胰酶的表达

目的观察豚鼠过敏性休克死亡肺组织中类胰蛋白酶和胃促胰酶的表达情况,试图为过敏性休克死亡提供客观的诊断依据。方法健康豚鼠24只,随机均分为实验组和对照组,每组再分死亡即时组、冷藏48h组和冷冻7d组,每组4只。实验组将0.5mL人混合血清用生理盐水1∶10稀释,注射于豚鼠后掌皮内,致敏后3周以人混合血清1mL注入心腔诱发过敏性休克致死;对照组采用生理盐水代替混合血清。提取豚鼠心血及肺组织,应用免疫组化染色和图像分析技术观察类胰蛋白酶和胃促胰酶的表达情况。结果对照组豚鼠肺组织中类胰蛋白酶和胃促胰酶阳性细胞数量较少,分布在小血管和小气管周围。实验组肺组织中类胰蛋白酶和胃促胰酶阳性细胞明显增多,多数细胞形态不规则,阳性染色颗粒脱出肥大细胞并弥散到组织间隙。冷藏48h和冷冻7d的条件下对这两种酶的表达无明显影响。结论过敏性休克致死豚鼠肺组织中类胰蛋白酶和胃促胰酶的表达明显增强,在冷藏48h和冷冻7d内的条件下,可作为过敏性休克死亡的一项诊断依据。     

Differential decomposition patterns in charred versus un-charred remains

Although researchers have examined many aspects of fire modification, the rate and pattern of decomposition in charred remains have not been studied previously. This study utilized 48 domestic pigs, divided into 24 charred (head, neck, and limbs burned to Crow-Glassman level 1 and torso to level 2) and 24 un-charred pig carcasses. Decomposition of control carcasses was scored at 50 accumulated degree days (ADD) intervals, and charred carcasses were also observed and photographed at this time. A Charred Body Scale was subsequently created, and charred carcasses were scored retrospectively for the same ADD intervals. Analysis using a mixed-effect repeated measures model indicated that, while decomposition rate was not statistically different between the two groups (p = 0.2692), the charred remains initially displayed an ostensibly more advanced pattern. Body regions displaying significant charring decomposed at a faster rate (p < 0.001), while areas with very light levels of charring decomposed at a significantly slower rate (p < 0.001).