Erythrocytes in cervical lymph nodes of the human as a sequela of stasis and/or lymph drainage. Questionable diagnostic significance in strangulation and mechanical injuries of the head

Information on the diagnostic significance of follicular hemorrhage and/or presence of red blood cells in the lymph node sinus was obtained by microscopic investigation of the superficial and deep human cervical lymph nodes. Fifty cases were selected on the basis of the following criteria: (1) cases without strangulation or mechanical head trauma; (2) cases in which death occurred by strangulation without additional mechanical head trauma; (3) cases without strangulation but with mechanical head trauma; (4) cases with strangulation and mechanical head trauma. The usual degree of stasis and hemorrhages and the dilation of the veins and/or capillaries are not sufficient to discriminate between cases with and cases without strangulation. Moreover, erythrocytes, erythrophages, and siderophages were encountered in the lymph node sinus of both cases with and cases without mechanical head trauma. Neither follicular hemorrhage nor the presence of red blood cells in the sinus is therefore diagnostically significant in forensic pathology.     

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.     

不同死因大鼠死后不同时间红细胞溶血速度比较

目的观察不同死亡原因大鼠尸体血液中红细胞溶血速度的变化规律,为死亡原因的法医学推断提供新思路。方法 40只SD大鼠随机分为4组,分别以断髓、置于99%CO的空间、高坠、勒颈方式处死后,取各组大鼠右心室血液,于死后即刻（0h）、8h、16h、24h、32h、40h、48h、56h、64h、72h,采用显微镜数码图像法进行全血红细胞计数（CBC）,并对组内和组间数据进行统计学比较分析。结果 4组血液红细胞数量在死后即刻至72h期间,均随时间的延长因溶血而减少。其中0~16h,各组溶血速度无明显差异（P〉0.05）;16~48h,速度加快,溶血速度以CO中毒组（88.50±25.99）%最快,其次为高坠（69.33±29.52）%和断髓组（48.78±3.17）%,机械性窒息组（41.90±9.61）%最慢,组间比较具有显著性差异（P〈0.05）;56h后,溶血速度再次减慢,至72h机械性窒息组仍有少量红细胞存在。结论 4组不同死因大鼠死后不同时间红细胞计数均减少,各组间差异具有统计学意义,其变化特征可为死亡原因的推断提供新的思路。     

利多卡因在蛛网膜下腔和静脉注射致死犬体内的死后分布

目的比较利多卡因在蛛网膜下腔和静脉注射致死犬体内的死后分布特点。方法犬12只,其中6只经蛛网膜下腔,另6只经股静脉匀速注入利多卡因(5×15mg/kg)致死,迅速解剖动物,取大脑、侧脑室脑脊液、腰段脊髓腔脑脊液、不同脊髓节段(颈髓、胸髓、腰髓、骶髓),心、肺、肝、脾、肾、胆汁、尿、心血、周围血、注射部位肌肉和注射部位20 cm以外肌肉等脏器组织和体液,用气质联用法定性,气相色谱法定量检测其中利多卡因含量。结果蛛网膜下腔注射致死犬体内利多卡因的含量由高到低顺序依次为腰段脊髓腔脑脊液、骶段脊髓、胸段脊髓、侧脑室脑脊液、腰段脊髓、颈段脊髓、肺、肾、注射部位肌肉、心、大脑、脾、心血、肝、周围血、胆汁、注射部位20 cm以外的肌肉、尿;静脉注射致死犬体内利多卡因的含量由高到低顺序依次为肾、心、肺、脾、大脑、肝、周围血、胆汁、心血、颈段脊髓、胸段脑脊液、注射部位肌肉、腰段脊髓、注射部位20 cm以外的肌肉、侧脑室脑脊液、尿、腰段脊髓腔脑脊液、骶段脊髓。结论蛛网膜下腔注射致死犬背侧脊髓液中利多卡因含量最高,静脉注射致死犬肾脏利多卡因含量最高,此分布特征可为利多卡因麻醉意外法医学鉴定中入体途径的判定提供参考。     

尸体心血红细胞ATP降解规律与早期死亡时间的相关性

目的观察早期死亡时间（PMI）与血液红细胞ATP含量的相关性。方法选择具有确切死亡时间的尸体30例,在死亡后6、8、10、12、14、16、18、20、22、24h分别于第4肋间进行心脏穿刺取血,利用生物发光法检测血液样本红细胞ATP含量（μmol／gHb）,并观察红细胞ATP含量变化与死亡时间的关系。结果尸体心血红细胞ATP含量在死亡后1～24h之内呈现非匀速下降趋势,与死亡时间的Pearson相关系数为-0．971（P=0．000）;尸体心血红细胞ATP含量与死亡时间的回归方程及尺。值为：Y=-0．096X＋2．872（x为死亡时间）,R2=0．936,P=0．000。结论尸体心血红细胞ATP含量在死后1—24h之内的变化与死亡时间具有相关关系,可以作为法医学死亡时间推断的生物学指标。     

Age determination of blood spots in forensic medicine by force spectroscopy

We present a new tool for the estimation of the age of bloodstains, which could probably be used during forensic casework. For this, we used atomic force microscopy (AFM) for high-resolution imaging of erythrocytes in a blood sample and the detection of elasticity changes on a nanometer scale. For the analytic procedure we applied a fresh blood spot on a glass slide and started the AFM detection after drying of the blood drop. In a first step, an overview image was generated showing the presence of several red blood cells, which could easily be detected due to their typical "doughnut-like" appearance. The consecutively morphological investigations in a timeframe of 4 weeks could not show any alterations. Secondly, AFM was used to test the elasticity by recording force-distance curves. The measurements were performed immediately after drying, 1.5 h, 30 h and 31 days. The conditions were kept constant at room temperature (20 degrees C) and a humidity of 30%. The obtained elasticity parameters were plotted against a timeline and repeated several times. The elasticity pattern showed a decrease over time, which are most probably influenced by the alteration of the blood spot during the drying and coagulation process. The preliminary data demonstrates the capacity of this method to use it for development of calibration curves, which can be used for estimation of bloodstain ages during forensic investigations.     

Pulmonary siderophages and unexpected infant death

It has been proposed that the presence of siderophages in the lungs of infants who die unexpectedly should be considered a marker of a previous hypoxic event, which may preclude a diagnosis of sudden infant death syndrome. The authors retrospectively reviewed all infant deaths (<1 year old) going to autopsy at the Denver Office of the Medical Examiner from January 1999 to January 2001. Lung sections were stained with Prussian blue, and siderophages were counted in 20 high-power fields per lobe sampled. Cell counts were performed by two independent pathologists who were blinded to history and cause of death, with good reproducibility. Iron stain results were then categorized by average number of siderophages per 20 high-power field (category 1 = <5, category 2 = 5-100, category 3A = 100-500, category 3B = >100 in a single lobe, category 4 = >500). The results were subsequently correlated to case history, autopsy findings, and cause/manner of death. Forty-three cases were reviewed. The causes of death included sudden infant death syndrome (16), asphyxia (5), undetermined (6), and other (16). Those deaths were categorized by the above criteria as follows: category 1. (32), category 2. (6), category 3. (4), and category 4. (1). All sudden infant death syndrome deaths were in category 1. Categories 1 and 2 also included deaths in which hypoxia might have been present before death because of such factors as pneumonia and congenital heart disease. Categories 3 and 4 included a known homicidal asphyxia in which repeated episodes of intentional smothering were documented, 2 probable asphyxias, 1 nonaccidental trauma, and 1 undetermined. All 5 cases had questionable circumstances surrounding the death of the infant. Pulmonary siderophages were described in only 1 of the 43 autopsy reports. It was concluded that pulmonary siderophages can be markedly increased in cases of repeated asphyxia. Siderophages may also be increased in cases where hypoxia may have been present for another reason, but not to the same degree. Siderophages are not increased in sudden infant death syndrome. Because iron-laden macrophages often are not recognized on routine examination with hematoxylin and eosin staining, iron stains may be helpful in the evaluation of infant deaths. If siderophages are present in increased amounts without an obvious explanation, further investigation is warranted.     

Postmortem stability of DNA

High-molecular-weight DNA was recovered postmortem in sufficient quantities from various human organ tissues as well as from blood, although not all organs were equally well suitable. Good DNA stability was found in brain cortex, lymph nodes and psoas muscle over a period of three weeks postmortem. Spleen and kidney showed good DNA stability up to five days postmortem but after longer periods, rapid degradation was observed. Yields of DNA from blood were not consistent because of the non homogeneity of samples. Blood clots were rich with DNA. Generally, the amount of degraded DNA correlated directly with the duration of the postmortem period. However in some cases, DNA degradation was already prominent after a short period. However in some cases, DNA degradation was already prominent after a short period. Case histories showed that high environmental temperature at the site of death and/or infectious diseases prior to death were the main factors for rapid autolysis. Gradual disappearance to complete loss of the long fragments (15-23 kb) was observed in DNA fingerprinting using the minisatellite probe 33.15. No extra-bands were noted, thus excluding erroneous conclusions. However, evidentiary value of older samples was lower.     

抗人N单克隆抗体的制备及其特性的研究

用ON型人红细胞免疫BALB/c小鼠脾细胞与SP2/O骨髓瘤细胞融合,获得分泌IgM类抗人N单克隆抗体杂交瘤细胞株(ON_1D_3)。其培养上清液抗体效价为128～256,诱生小鼠腹水的抗体效价为8192;与Panel A红细胞反应呈抗N特异性;检测273名成人红细胞,只特异性凝集N(?)MN型人红细胞,不凝集M型红细胞。与猴等9种动物红细胞无交叉反应;酶修饰试验结果表明,ON_1D_3识别的抗原决定簇是唾液酸依赖性的。     

Morphine and 6-acetylmorphine concentrations in blood, brain, spinal cord, bone marrow and bone after lethal acute or chronic diacetylmorphine administration to mice

The aim of this study was to evaluate postmortem incorporation of opiates in bone and bone marrow after diacetylmorphine (heroin) administration to mice. Mice were given acute (lethal dose of 300 mg/kg) or chronic (10 and 20 mg/kg/24 h for 20 days) intraperitoneal administration of diacetylmorphine. The two metabolites of diacetylmorphine, 6-acetylmorphine (6-AM) and morphine, were extracted from whole blood, brain, spinal cord, bone marrow and bone (after hydrolysis) using a liquid/liquid method. Quantification was performed by gas chromatography-mass spectrometry (GC/MS). Results showed that after acute administration, opiates were present in all studied tissues. Morphine concentrations appeared to be higher than those of 6-AM in blood (52.4 microg/mL versus 27.7 microg/mL, n=12), bone marrow (87.8 ng/mg versus 8.9 ng/mg, n=6) and bone (0.85 ng/mg versus 0.43 ng/mg, n=6), but 6-AM concentrations were higher than those of morphine in brain (14.0 ng/mg versus 7.4 ng/mg, n=12) and spinal cord (27.8 ng/mg versus 20.8 ng/mg, n=12). No correlation was found for both compounds between blood concentrations and either brain, spinal cord, bone or bone marrow concentrations while a significant one was found between brain and spinal cord concentrations either for morphine (r=0.89, n=12, p<0.001) or 6-AM (r=0.93, n=12, p<0.001), the concentration being higher in spinal cord than in brain. When bones were stored for 2 months, only 6-AM remained in bone marrow but not in bone. After chronic administration, mice being sacrificed by cervical dislocation 24 h after the last injection, no opiate was detected in any studied tissues. Further studies are required, in particular in human bones, but these results seem to show that 6-AM could be detect in bone marrow several weeks after the death and could be an alternative tissue for forensic toxicologist to detect a fatal diacetylmorphine overdose, even if no correlation between blood and bone marrow was observed. On the other hand, neither bone tissue nor bone marrow will allow the confirmation of a chronic diacetylmorphine use.     

迟发性外伤性脑内血肿脑组织GFAP、S-100和NSE的免疫组织化学研究

应用免疫组织化学ABC法和S－P法，对6例伤后1．5天至2年发生的迟发性外伤性脑内血肿（DTICH）脑组织标本进行神经胶质细胞内胶质纤维酸性蛋白（GFAP）、S－100蛋白（S－100）和神经元细胞内特异性烯醇化酶（NSE）变化和应用价值的研究。尸检和组织学检查发现有脑挫伤等病变。6例DTICH脑组织挫伤及周围区均有明显的GFAP、S－100阳性的细胞变化，细胞数目增多，胞体肥大、增生，免疫组化反应产物增多、深染；神经元细胞NSE免疫组化染色均呈阴性改变。结果表明：6例DTICH脑组织中GFAP、S－100的NSE的免疫组织化学变化是有一定时间间隔的外伤性陈旧性改变，在DTICH的病理诊断上具有重要价值，可作为DTICH法医学鉴定的一个辅助手段。     

实验性大鼠脑震荡后c-jun mRNA表达

目的观察实验性大鼠脑震荡后c-jun mRNA的表达变化规律。方法 55只实验大鼠随机分为脑震荡组（0min、15min、30min、60min、3h、6h、12h、24h、48h、96h）和对照组,用原位杂交法观察大鼠脑震荡后各时间点,大脑皮质、脑干和丘脑神经元c-jun mRNA表达的变化规律。结果对照组大鼠神经元和胶质细胞均可见c-jun mRNA的弱阳性表达。脑震荡组大鼠损伤后15min神经细胞观察到c-jun mRNA阳性表达,随损伤后经过时间的延长阳性表达逐渐增强;30min时c-jun mRNA阳性反应达高峰,随后逐渐降低,至96h时回落至对照组水平。结论 c-jun mRNA表达水平可成为诊断脑震荡和推断伤后经过时间的一项敏感指标。     

Autolytic changes in blood cells and other tissue cells of human cadavers. I. Viability and ion studies

The viability of white blood cells, spermatozoa of the epididymis, cells from minced spleen, lymph node and lung, and cells aspirated from the bone marrow of the sternum was studied using the vital dye exclusion test. The material comprised 123 medicolegal autopsy cadavers which had been stored in a mortuary cold room at +4 °C up to 10 days after death. Cells which excluded trypan blue were found in various specimens from all cadavers. However, there was marked individual variation in the results. The loss of viability of the white blood cells showed a moderate correlation (r = ?0.78) with the increase in the post-mortem (p.m.) time, whereas the results for other tissues were not so significant. The K⁺ and Mg²⁺ and haemoglobin content and the osmotic resistance of the red cells correlated poorly with the p.m. time. The present results suggest that despite the general assumption that autolytic changes proceed rapidly at the cellular level, individual cells and especially blood cells may remain viable for a long time in cadavers kept at +4 °C.     

Validation of Large White Pig as an animal model for the study of cannabinoids metabolism: application to the study of THC distribution in tissues

This study presents a new animal model, the Large White Pig, which was tested for studying cannabinoids metabolism. The first step has focused on determination of plasma kinetics after injection of Delta(9)-tetrahydrocannabinol (THC) at different dosages. Seven pigs received THC by intravenous injections (50, 100 or 200 microg/kg). Plasma samples were collected during 48 h. Determination of cannabinoids concentrations were performed by gas chromatography/mass spectrometry. Results showed that plasma kinetics were comparable to those reported in humans. Terminal half-life of elimination was 10.6 h and a volume of distribution of 32 l/kg was calculated. In a second step, this model was used to determine the kinetic profile of cannabinoids distribution in tissues. Eight Large White male pigs received an injection of THC (200 microg/kg). Two pigs were sacrificed 30 min after injection, two others after 2, 6 and 24 h. Different tissues were sampled: liver, kidney, heart, lung, spleen, muscle, fat, bile, blood, vitreous humor and several brain areas. The fastest THC elimination was noted in liver tissue, where it was completely eliminated in 6 h. THC concentrations decreased in brain tissue slower than in blood. The slowest THC elimination was observed for fat tissue, where the molecule was still present at significant concentrations 24 h later. After 30 min, THC concentration in different brain areas was highest in the cerebellum and lowest in the medulla oblongata. THC elimination kinetics noted in kidney, heart, spleen, muscle and lung were comparable with those observed in blood. 11-Hydroxy-THC was only found at high levels in liver. THC-COOH was less than 5 ng/g in most tissues, except in bile, where it increased for 24 h following THC injection. This study confirms, even after a unique administration, the prolonged retention of THC in brain and particularly in fat, which could be at the origin of different phenomena observed for heavy users such as prolonged detection of THC-COOH in urine or cannabis-related flashbacks. Moreover, these results support the interest for this animal model, which could be used in further studies of distribution of cannabinoids in tissues.     

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.     

常规冷冻对大鼠尸体组织形态学的影响

目的观察分析死后不同时间冷冻大鼠尸体主要内脏组织细胞形态变化规律。方法 SD大鼠20只,分为尸僵前冷冻组、尸僵期冷冻组、尸僵缓解冷冻组及对照组（各5只）,颈椎脱臼处死大鼠,实验组大鼠在室温25℃±5℃、相对湿度80%±5%分别放置0h、6h、24h后-20℃±2℃冰箱冷冻72h,解剖提取脑、心、肺、肝、肾,常规切片HE染色,显微图像体视学测量细胞外间隙的面密度。结果各实验组内脏组织间质疏松增宽,冰晶裂隙沿组织间隙或肝血窦分布、周边细胞固缩,红细胞冻溶、血管周围浆液淤积,以肝组织变化最明显。细胞外间隙的面密度：对照组明显小于实验组（P〈0.01）;脑尸僵前组较尸僵组大（P〈0.05）;心、肾尸僵前组和尸僵组无显著差异（P〉0.05）,均较尸僵缓解组大（P〈0.01）;肝各实验组组间比较均无显著差异（P〉0.05）。结论冷冻不同尸僵状态下内脏组织细胞有不同的形态结构,可作为区别和判断生前和死后组织细胞形态改变的参考。     

Immunohistochemical research on brain stem injury: expression of heat shock protein 70 in rat brain after traumatic brain stem injury

The aim of this study was to determine the expression of heat shock protein 70 (HSP70) as molecular chaperones, following traumatic brain stem injury(TBSI). The expression of HSP70 was detected in respective brain regions of rats 1 h, 3 h, 6 h, 12 h, 24 h after brain stem stabbing wound. Numerous HSP70 immunoreactivity positive blood vessel endothelioid and glia cells were found in cerebral, cerebellar cortex and hippocampus after 1 h, with H.E stain unchanged. In the area near the focal of brain stem injury, HSP70 immunoreactivity positive neurons increased significantly after 1 h of injury, reaching the highest level after 3 h, much higher than in other areas. HSP70 immunoreactivity positive cells still existed after 24 h of injury. It was suggested that TBSI cause the defectiveness of protein structure in brain, and induce HSP70 proteolysis of denature protein including misfolding or aggregate. TBSI can be diagnosed regionally by the increasing of HSP70 immunoreactivity positive neurons in the focal of brain stems.     

Sudden Collapse due to Medullary and Cervical Cord Infarction—An Unusual Presentation of Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is a common cancer worldwide with a great potential for metastatic spread. Hepatocellular carcinoma often arises in people with underlying viral hepatitides or liver cirrhosis and may present in various ways including abdominal pain, liver mass, and signs of hepatocellular decompensation. Many tumors may have metastasized to other organs such as the lungs, lymph nodes, bone, and adrenal glands at the time of diagnosis. However, it is uncommon for HCC to present purely due to its metastasis, such as spinal cord compression from vertebral metastasis. Here, an unusual presentation of a sudden cardiovascular collapse due to medullary and cervical cord infarction from compression of the cervical cord is presented. The importance of clinical investigations, the usefulness of postmortem computed tomography scans, and the examination of the cervical spine and cervical cord in people with no obvious cause of death after standard autopsy procedures are emphasized.     

大鼠脑外伤后溶酶体酶Cathepsin—B和D的表达

目的研究大鼠脑外伤后溶酶体酶cathepsin-B和-D是否被激活及其不同时段表达变化,阐述其与凋亡执行因子caspase-3表达的关系,并探讨对脑外伤诊断及形成时间的意义。方法采用自由落体打击法建立脑外伤动物模型,并对模型及对照样本进行免疫荧光、双标和激光共聚焦检测,结果用SPSS10.0软件处理。结果脑外伤后1hcathepsin-B表达即增加,4～8d达高峰,脑外伤后32d仍处于高表达水平;cathepsin-D的表达于脑外伤后12h增加,4～8d达高峰,32d的表达仍然高于12h的表达水平。脑外伤初期,cathepsin-B和-D阳性细胞与caspase-3阳性细胞重叠较少,脑外伤后6h开始增加,32d仍然有很多阳性细胞重叠。结论脑外伤后cathepsin-B和-D被激活,其激活在脑外伤早期可能抑制细胞凋亡执行因子caspase-3的激活,之后(6h后)则与caspase-3起协同作用,共同促进细胞死亡;cathepsin-B和-D表达的时程变化对于脑外伤的法医学诊断和中晚期的时间推断有参考意义。     

小鼠各脏器及血中甲基苯丙胺的分布

目的通过给小鼠腹腔注射甲基苯丙胺(MAMP),建立小鼠急性染毒模型。方法运用GC/NPD技术,系统地进行了染毒小鼠模型各脏器及血中甲基苯丙胺(MAMP)的定性定量测定。采用1-萘胺作为内标,样品处理时采取加盐酸成盐后,又碱化,用有机溶剂萃取浓缩挥发。结果回收率高于80%。血、各脏器中最低检出限为2～5ng/ml。结论脑、肾、脾组织可以用于法庭科学分析。