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.     

Genotyping of DNA samples under adverse conditions of Low Copy Number—LCN (formolisados tissue samples and embedded in paraffin)

This paper aims to describe and evaluate a protocol for extraction of DNA (deoxyribonucleic acid) in formalinized tissues and embedded in paraffin for forensics genetic analysis. In outline the method is the removal of paraffin with an organic solvent in 0.3–0.5 mg of the sample of the tissue under study, followed by removal of formaldehyde, rehydration and soon after the extraction of genomic DNA. The extraction is achieved through the stages of cellular lysis, enzymatic digestion of proteins and DNA precipitation in ethanol medium. With the research we can conclude that even when the DNA is present in small quantities in conditions of extreme difficulties in its extraction, as formalinized tissues and embedded in paraffin, the technique of optimizing the extraction of DNA used both to organic extraction as Chelex, for use in the polymerase chain reaction (PCR), and possible the investigation of different samples of human tissue, biological samples, or was obtained under the conditions tested, a DNA with good quality and concentration. The samples were amplified for the mini-STRs loci using the product marketed in multilocus, using a methodology recommended by the supplier and validated for analysis of forensic DNA. Commercial kit was used MiniFiler from Applied Biosystems. The DNA fragments amplified by PCR showed that the extracted DNA had good amplification.     

甲醛固定石蜡包埋组织STR分型检测

目的评估10%甲醛固定石蜡包埋组织STR分型结果的影响因素。方法采用QIAGEN法、IQ法、Chelex法对2具新鲜尸体在尸检时常规制备的心、脑、肝、脾、肾、肺、胃、肠石蜡包埋组织进行DNA提取,用AmpFlSTR Identifiler试剂盒进行PCR扩增,在3100-Avant上完成片段分析。另外对15个案例中室温保存1～5年的心、肝、肺、肠存档石蜡包埋组织共56份采用同样的方法进行STR分型。以STR基因座检出率评估分型有效性。结果各种组织DNA片段均随着保存时间延长而持续降解,其中心、肺组织STR基因座检出率与保存时间存在线性相关。相同保存时间时,各种组织基因座检出率差异有统计学意义。其中以肺组织在不同保存时间中基因座检出率最高。结论在甲醛固定时间一定的条件下,存放时间、组织类型、DNA提取方法和PCR模板质量浓度是影响石蜡包埋组织STR分型的重要因素。     

福尔马林固定石蜡包埋组织中DNA提取

由于甲醛介导的DNA损伤和石蜡对DNA提取的阻碍作用,使得常规DNA提取方法很难从福尔马林固定石蜡包埋组织(FFPET)中获取高质量的DNA。近年来,众多学者的研究表明,通过改良预处理方法,优化蛋白酶K消化作用,简化DNA提取步骤,纯化DNA提取物等,可以有效提高FFPET中提取的DNA质量,为FFPET的DNA分析奠定了基础。     

Mechanical airway obstruction caused by accidental aspiration of part of a ballpoint pen

The authors present three cases of death in children aged 4, 9, and 10 years, respectively, that were first thought to be caused by herbal or other poisonings but at autopsy were found to be caused by airway obstruction from aspiration of ballpoint pen parts. Aspiration of a foreign body is a leading cause of accidental death in children, but the circumstances in these cases were unique. In the first case, a 4-year-old child died shortly after a visit to a traditional healer. The child's mother blamed him for the death and fatally assaulted him. The second case was a 9-year-old who died at school. Case 3 was a 10-year-old who collapsed while playing with a ballpoint pen in her mouth. In the latter two cases, the relatives alleged poisoning. At autopsy, there was no evidence of trauma, disease, or poisoning in all three cases. Ballpoint pen parts were present in the larynx, carina, and left main bronchus, respectively. Features of "asphyxial" death were present, and included subconjunctival hemorrhages, subendocardial hemorrhages, and congestion of the face and internal organs. These deaths are preventable by education of children, parents, and teachers. Ballpoint pen manufacturers should also modify the design of these pens to improve their safety.     

Allelic alterations of STRs in archival paraffin embedded tissue as DNA source for paternity testing

Owing to a wrong name registered on ID card, the identity of a businessman who had been dead and cremated was suspected, which led his son failed to get legacy. In order to prove the parenthood, the son submitted the gastric cancer tissues surgically removed and embedded in a paraffin block as DNA source for paternity test. After extracting DNA with QIAamp DNA Blood Mini Kit, the 16 STR loci was amplified by two commercial kits of Sinofiler^® (ABI)and Powerplex 16 (Promega), respectively. Both of the STR profiles were similarly showing allelic imbalance pattern at some loci and an additional allele at locus D18S51. The cancerous tissues and adjacent normal tissues were then partitioned off from each other by microscopic analysis of H.E. stained sections and followed by DNA extracting and STR typing, respectively. The allelic alteration could not be found in normal tissues whereas it did in cancerous tissues whose STR profile showed complete loss of one allele (LOH) at loci D13S317 (allele 11 was lost), partial loss of one allele (pLOH) at loci D21S11, D7S820, D19S433, vWA, D12S391 and Amelogein and occurrence of an additional allele (allele 20 was added) at locus D18S51. The results demonstrated that the Paraffin Embedded cancer Tissue used as DNA source for forensic identification is possibly questionable because of their microsatellite instability (MSI) or loss of heterozygosity. It was suggested to partition the normal tissues from the cancer tissues by microscopic evaluation first and then analyzing DNA separately. Comparing the STRs profile of normal tissue with the son's blood sample, the final conclusion was acquired that the donor of the paraffin embedded tissues is the biological father of the son.     

Preventing paraffin-related injury

Paraffin (called kerosene in North America and other parts of the world) is the most commonly used fuel in ?non-electrified dwellings worldwide. It is especially popular in Africa and South Asia. Although paraffin ?offers many advantages-especially its comparatively low cost to produce-it poses two major risks of ?injury. First, paraffin poisoning is common, either through ingestion or through inhalation of smoke and ?fumes. Second, paraffin is highly flammable, and poses fire risk through multiple causes. This commentary ?discusses strategies to prevent paraffin-related injury. Prevention of paraffin-related injury must be through ?multiple strategies, and should include policy-oriented change, changes to the safety of home environments, ?and behavioral changes targeting how individuals store and use paraffin and paraffin appliances. We review ?successful prevention strategies in each of these domains and discuss appropriate research and community ?initiatives that should be implemented to improve paraffin safety among at-risk populations. ?     

Multiple interchanging of tissue samples in cases of breast cancer

Due to the suspicion of a gynaecologist, a pathologist was suspected of incorrect diagnoses in cases of breast cancer and the interchanging of tissue samples. Many women applied to the attorney's bureau to clarify the reproaches. The privately owned laboratory for pathology was searched and 926 histological slides, roughly the same number of paraffin blocks and about 20 formalin fixed tissue samples were confiscated. Together with other confiscated material, at least 1236 histological slides and additional 249 paraffin blocks had to be sorted. Histological slides and paraffin blocks were matched with patients as far as possible following the laboratory book. Many of the warranted samples which were diagnosed as containing the carcinoma by the pathologist were missing. A total of 160 samples were chosen and rediagnosed by two independent pathologists. The formalin fixed tissue was negative for DNA most likely due to storage in formalin for years. Most of the histological slides were positive for DNA. On the whole, 18 expertises about histological findings and the DNA results were given. In some cases only DNA results could be presented, as previous experts had only performed DNA examinations without controlling the histological diagnosis. In six cases a carcinoma could be confirmed and the DNA profile matched with patient's DNA; in seven cases a carcinoma was confirmed without match with the patient; in two cases the carcinoma could not be confirmed in the presented samples. A jurisdictional solution was impossible because the accused pathologist died during the investigation. In conclusion, it must be stated that a DNA examination of histological slides should never be performed without a rediagnosis of an independent pathologist and photographic documentation of the findings. Whenever possible, material should be left on the slide.     

Self‐Embedding Behavior in Adults: A Report of Two Cases and a Systematic Review

Self‐embedding behavior (SEB) is the repeated insertion of sharp objects, such as needles or pins, into the soft tissues of abdomen, limbs, and other body parts. In this study, two cases of SEB were reported and the scientific worldwide literature reviewed. Thirty‐two cases of SEB were identified through systematic searches in the main bibliographic databases. Mean age was 35 years (SD = 8.97). Just over two‐thirds of the patients were female. Although the number of embedded objects could be as high as 200, major clinical and surgical complications were uncommon and mortality was null. Patients with SEB presented three major diagnoses: psychotic (25%), personality (21.9%), and factitious (28.1%) disorders. The practice of SEB largely went undetected as the patients themselves did not bring it to the attention of family members or physicians and usually denied they have engaged in SEB. A high level of suspicion is required to avoid a missed diagnosis.     

Changes in the ratio of collagen type I and III in the interlobular interstitium of the lung in sudden infant death--a pilot study

In a blind study the interlobular interstitial connective tissue in paraffin embedded lung sections from 45 autopsy cases aged under 2 years (23 SIDS and 22 Non-SIDS) were investigated with the aim of determining the collagen type I:type III ratio by means of polarimetric evaluation. Sections were stained with Resorcin-Fuchsin for elastin fibres, Celestine Blue/Mayer's Haematoxylin for nuclear details and with Solophenyl Red 3 BL in saturated picric acid solution for the differential staining of type I and type III collagen fibres for polarization microscopy. The type I fibres were orange and the type III fibres green in colour. The spectral distribution of the coloured polarized light from the sections was determined and peaks were evident at the wavelengths 590 nm for the orange coloured light of type I and 490 nm for the green coloured light of type III collagen. With corresponding filtres the intensities of the orange and green emissions were separately measured at several points adjacent to lymphatic vessels. The ratio collagen I/III, deduced from the ratio of the intensities of orange to green light, was significantly higher in the SIDS-group than in the Non-SIDS-group (a = 0.001) due to the increase in the amount of collagen type I and could indicate an insiduous fibrosis resulting from lymphostasis.     

FN-EIIIA肽段表达与皮肤损伤时间推断研究

目的 在mRNA研究的基础上,研究大鼠皮肤挫伤后含EIIIA□段的Fn蛋白水平表达与损伤时间关系。方法片段特异性单克隆抗体免疫组织化学染色。结果伤后6h表皮细胞、毛囊上皮细胞等出现棕黄色阳性染色颗粒,并随损伤时间延长染色加深。结论Fn-EIIIA片段检测可用于法医学生前伤确认和损伤时间推断。     

吗啡类毒品中毒死者主要脏器内吗啡分布的研究

目的研究吗啡在人体脏器中的分布情况,筛选适于免疫组化方法检测吗啡的脏器。方法用免疫组化SP法对8例吗啡类毒品中毒死亡尸体脑、肾、心、肝等脏器中的吗啡进行定位和半定量观察。结果吗啡主要分布在某些实质细胞的胞浆内,肾脏和脑组织的吗啡含量高,不同器官、不同案例吗啡的分布情况差异较大。结论肾脏和脑是实际检案时用免疫组化方法检测吗啡的理想检材。     

Ethical criteria for procuring and distributing organs for transplantation

This article provides an ethical analysis and assessment of various actual and proposed policies of organ procurement and distribution in light of moral principles already embedded in U.S. institutions, laws, policies, and practices. Evaluating different methods of acquisition of human body parts--donation (express and presumed), sales, abandonment, and expropriation--the author argues for laws and policies, including required request, to maintain and facilitate express donation of organs by individuals and their families. Such laws and policies need adequate time for a determination of their effectiveness before society moves to other major alternatives, such as a market. In organ allocation and distribution, which have close moral connections with organ procurement, the author defends the judgment of the federal Task Force on Organ Transplantation that the community should have dispositional authority over donated organs, that professionals should be viewed as trustees and stewards of donated organs, and that the public should be heavily involved in the formation of policies of allocation and distribution. Concentrating on policies being developed in the United Network for Organ Sharing, the author examines the point system for cadaveric kidneys, the access of foreign nationals to organs donated in the U.S., and the multiple listings of patients seeking transplants. He concludes by identifying two major problems of equitable access to donated organs that will have to be addressed by social institutions other than UNOS: access to the waiting list for donated organs and the role of ability to pay in extrarenal transplants.     

An improved method for the histological preparation of single hairs and dust samples--morphological and immunological examination

An improved method of the serological and morphological investigation of human hair is reported. The hair was firmly fixed onto a microscopic slide with cellophane tape and observed microscopically to confirm the cuticula images and the presence of the medulla. A piece of the hair containing the medulla was dissected, embedded in paraffin, and a cross section of this hair was prepared. By treating the sample with immunohistochemistry (biotin-antibiotin ABC technique), the blood type of the hair was confirmed definitively. Dust containing shaved beard can be examined in the same way.     

Diagnosis of stress preceding the fatal outcome

Postmortem examinations of cytomedines of internal organs in 181 men showed a possibility to use their (cytomedines') quantitative and qualitative characteristics as diagnostic criteria of stress influencing the human organism shortly before the death onset. Bioregulatory peptides were obtained by using Morozov's and Khavinson's method of acetous extraction. The composition of cytomedines was determined by spectrophotometer, while their biological activity (BA) was studied through affecting the phagocytic activity of blood neutrophils. It was concluded that, under stress, there are a significant inhibition of BA and a reduced count of cytomedines in the internal organs. The found data can be used to detect the stress preceding death.     

Use of scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDXA) in identification of foreign material on bullets

The authors report two cases in which examination of foreign material embedded in or adherent to bullets provided critical information in the reconstruction of a crime scene. Analysis of small particles by scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDXA) can be accomplished without destruction or injury of the particles. In one case, the detection and identification of mineral fragments embedded near the nose of a bullet provided conclusive evidence that the bullet had ricocheted from a fireplace before striking the victim. In the second case, analysis of particles from two bullets identified them as them as bone fragments, thus proving which shots fired from a police officer's gun had killed a suspected burglar. SEM-EDXA has not been widely used to identify such material on bullets, but should be considered a potentially powerful tool in forensic science.     

红外显微光谱技术分析陈旧性心肌梗死酰胺特征

目的应用红外显微光谱技术分析陈旧性心肌梗死组织中酰胺Ⅰ、Ⅱ、Ⅲ含量及相互关系,以探讨陈旧性心肌梗死过程中蛋白质分子的变化。方法选取20例冠心病猝死案例(均含陈旧性心肌梗死病变)的心脏样本,在每个心脏心肌梗死边缘处取材2块,常规固定、制作蜡块、连续切片2张,其中1张行HE染色,经Dotslide 2.1工作站扫描设定陈旧性心肌梗死区为B组(病变区),邻近非梗死病变区为A组(非病变区);另1张切片常规脱蜡、脱水,在扫描图像指导下,于相应区域行红外显微光谱微区检测,应用OMNIC 8.0工作站处理、分析光谱。结果与A组相比,B组显微光谱示酰胺Ⅰ、Ⅱ吸收峰的变化趋势一致,均为峰位蓝移、峰增宽、峰强度增加;酰胺Ⅲ吸收峰为峰位蓝移、峰增宽、峰强度降低;两组比较差异具有显著性意义(P<0.05,P<0.01);B组酰胺Ⅰ峰高/酰胺Ⅱ峰高增加(P<0.01)。结论与非病变区域心肌组织相比,陈旧性心肌梗死组织的蛋白质含量及构成均有差异,该病变过程存在氧化应激作用影响。     

Determination of heavy metals in some human organs, tissue, and fluids under normal conditions

A table has been created, presenting normal levels of heavy metals (Hg, Cd, Cr, Cu, Fe, Mn, Pb, Ni, Zn, Sn, Tl, Ag) in human biological media. In contrast to other handbooks, the authors use the normal values derived on the basis of their own investigations at the spectral laboratory of Bureau of Forensic Medical Expert Evaluations, Public Health Committee of Moscow and published reports. Comparative analysis was carried out for each element in human organs, tissues, and urine, which will facilitate expert work. Every laboratory engaged in expert evaluations and analysis in cases with suspected poisoning with heavy metal salts is to have reference values of these elements' concentrations in human organs and tissues obtained in this very laboratory on available equipment with consideration for specific features of its biogeochemical region.     

Identification and significance of delta-aminovaleric acid in putrefaction materials (author's transl)]

During former putrefaction experiments regularly a proteogenic substance has been found which by means of modern analytical methods now was identified as delta-aminovaleric acid (DAVA). DAVA seems to appear in guinea pig as well as human organs and some body fluids under experimental conditions never before the 3rd (20 degrees C) to 5th day (10 degrees C). It is characterized by statistically significant increases until the end of the 2nd (20 degrees C) to 5th week (10 degrees C) and relatively stable values thereafter. Considering storage temperature measurement of DAVA concentration can be of relevance for the estimation of the time of death in cases of putrescent corpses.     

刑事裁判权主体初论

刑事裁判权的主体是裁判机关,但其又依靠具体裁判组织行使职权。我国法院与裁判组织设置方面存在法院地方化、审判委员会裁判案件、陪审员虚置等问题,笔者提出的完善建议是:1.将法院的人事权和财政权收归法院系统,依据司法管区设置地方法院;2.调整审判委员会职权,取消其裁判重大疑难案件的权力,并建立同级法官会审制度;3.陪审员应当有权参与庭前审查工作,并针对部分案件设立陪审团制,使其与原来的参审制形成优势互补。