兔死后肝细胞自溶的超微结构观察

用透射电镜观察兔死后肝细胞的超微结构变化,发现：在0．5h即有轻微的自溶改变,表现为细胞核中异染色质异常凝集和滑面内质同扩张;在1h自溶变化已趋明显,且随死后时间延长,在2h、3h、4h、5h该变化越来越广泛而显著,可见线粒体肿胀,嵴断裂、消失,基质内絮状致密体（Flocculentdensehadies,FDB）形成,粗面内质网及核周池扩张等改变。肝细胞自溶的超微结构变化与死后时间相关,这对推断早期死亡时间有应用价值。     

Histopathological changes in selenium-exposed fish

Redear sunfish (Lepomis microlophus) were collected from Martin Lake in east Texas. For at least 8 months, 1 year earlier, aqueous selenium-laden effluent from man-made sources was released into this 5,000-acre reservoir (unpublished data). Redear sunfish from a reference lake, 8 km upstream, were collected for comparison to Martin Lake fish. The hepatopancreas (i.e., liver and associated, disseminated exocrine pancrease), mesonephros (i.e., kidney), gonads, heart, spleen, stomach, and gill arches were preserved for histopathological examination using optical and/or transmission electron microscopy. Livers from Martin Lake redear sunfish (which had accumulated approximately 20 ppm selenium in the liver) showed central necrosis, reduced quantities of rough endoplasmic reticulum and glycogen particles, and increased numbers of lysosome-like structures. Kidneys showed proliferative glomerulonephritis, and exocrine pancreas showed marked hypertrophy at the optical level. Ultrastructurally, architectural disorganization, reduced rough endoplasmic reticulum, increased cisternal space, and proliferation of smooth endoplasmic reticulum were evident. The stomach, spleen, gill, heart, and gonads showed no abnormalities.     

Effects of processing techniques on the forensic DNA analysis of human skeletal remains

Human remains processed by forensic anthropologists may potentially be used for genetic analysis. Therefore, the condition of the deoxyribonucleic acid (DNA) in processed remains may become an issue for future analysis. Processing techniques employed by anthropologists are highly variable and scanning electron microscopy reveals significant alterations to the bone surface depending upon the technique used. Such damage to the bone indicates differences may exist in quality and quantity of DNA extracted. This study assessed how five processing procedures used by major forensic anthropology laboratories around the country affects the amounts of DNA extracted from human rib bones and the subsequent DNA analysis. The DNA was analyzed using the short tandem repeat (STR) locus CSF1PO and amelogenin. The findings indicate processing procedures used by forensic anthropologists do not adversely affect DNA analysis but prolonged exposure to heat during processing may decrease the yield of information from the DNA.     

海洛因成瘾者神经、内分泌及免疫系统超微病理变化研究

目的 研究海洛因成瘾者中枢神经、内分泌、免疫及生殖系统的超微结构变化。方法 应用透射电镜技术对 4例海洛因成瘾者中枢神经系统、内分泌系统、免疫系统以及生殖系统进行了观察。结果 神经系统表现为线粒体肿胀、嵴断裂、空泡变、内质网扩张、神经纤维数量减少、细胞器减少;免疫系统表现为淋巴细胞数量减少,胞浆成分减少,死亡细胞增多;内分泌系统表现为线粒体扩张,局部嵴断裂,内质网扩张;生殖系统表现为男性生精细胞减少,细胞器减少。女性未见卵泡结构。结论 海洛因滥用者的亚细胞超微结构呈急、慢性缺氧性变性、坏死改变。     

Full STR profile of a 67-year-old bone found in a fresh water lake

DNA extraction from and DNA typing of fresh water-exposed aged bone specimens poses a challenging task and is not very well examined. This study presents a new method to extract typable DNA from such problematic bone specimens. The procedure comprises low-heat drilling and cryogrinding, mild lysis conditions, and silica-column-based DNA cleaning. DNA quantity is assessed by quantitative PCR prior to short tandem repeat (STR) amplification. The procedure was employed with a 67-year-old tibia bone fragment recovered from a fresh water lake and succeeded to produce a full STR profile using the MPX-SP1 and MPX-SP2 mini-STR kits and a partial profile with 12 successfully amplified STRs using the Identifiler STR kit. The new method for the extraction of DNA from aged fresh water-exposed bone specimens presented herein was successfully applied to prepare DNA of sufficient quality and quantity to generate a full STR profile.     

Early epidermal changes in heat- and electrically injured pig skin. II. An electron microscopic study

In order to find methods applicable for disclosing electrical torture, pig skin was exposed to heat and electricity under controlled circumstances. Biopsies for electron microscopy were obtained immediately after exposure. In heat lesions the nuclei were slightly distorted, sometimes with broken nuclear membranes. The tonofilaments were clumped, intracellular oedema was present and cell membranes were ruptured between desmosomes. In electrical lesions the nuclei were usually enlarged with strongly condensed chromatin. Some nuclei were composed of fine, evenly dispersed granular material. The cytoplasm appeared homogeneous, in large magnification finely granular. Cell borders could sometimes be identified located in situ. In the stratum corneum, which appeared normal in heat lesions, single or several cells or large areas had an electron-dense appearance. The difference in ultrastructure of heat and electrical lesions makes it probable that electricity has a specific action on epidermal cells.     

Busting the myths: DNA typeability after 48 hours of boil

The aim of our study was to monitor the quality and quantity of DNA in bone samples that were boiled for 48 h. Bos taurus bone disks were sampled every hour for 48 h. The subsequent DNA analysis used multiple mitochondrial DNA (mtDNA) targets (100–700 bp) to evaluate the quality and quantity of the DNA extracted. The DNA extracted from bone disks remained typeable after boiling for 48 h. We have proven that DNA typing results can be obtained even after long-term boiling.     

The Correlation Between Skeletal Weathering and DNA Quality and Quantity*

Abstract: Mitochondrial DNA analysis of skeletal material is invaluable in forensic identification, although results can vary widely among remains. Previous studies have included bones of different ages, burial conditions, and even species. In the research presented, a collection of human remains that lacked major confounders such as burial age, interment style, and gross environmental conditions, while displaying a very broad range of skeletal degradation, were examined for both mitochondrial DNA (mtDNA) quality and quantity. Overall skeletal weathering, individual bone weathering, and bone variety were considered. Neither skeletal nor bone weathering influenced DNA quality or quantity, indicating that factors that degrade bone do not have the same effect on DNA. In contrast, bone variety, regardless of weathering level, was a significant element in DNA amplification success. Taken together, the results indicate that neither skeletal nor individual bone appearance are reliable indicators of subsequent mtDNA typing outcomes, while the type of bone assayed is.     

Paraquat myopathy: report on two suicide cases.

We describe two suicide cases in which old paraquat was ingested. In conjunction with lung involvement a pronounced degeneration was observed in skeletal muscle of one who died on the 14th day after the ingestion. The following sarcoplasmic or endoplasmic reticulum Ca2+ ATPase (SERCA) monoclonal antibodies were used for skeletal muscle fiber typing by an immunohistochemical method: NCL-SERCA1, reactive with type 2 fiber (fast-twitch), and NCL-SERCA2, reactive with type 1 fiber (slow-twitch). The examination revealed that the remarkably degenerated fibers belonged to type 1 muscle fibers. This case showed an abrupt increase of plasma CK levels (1796 mU/ml) on the fifth day after the ingestion. The authors presume that the damage to the skeletal muscle had occurred in this period. The degeneration of the muscle seemed to be attributable to the long retention of paraquat in the tissue because these findings were not observed in the other case who died on the fifth day. Paraquat-induced myopathy may develop in prolonged paraquat poisoning. The examination of CK levels in plasma will be useful for diagnosis of damage of skeletal muscle.     

Difficulties of sex determination from forensic bone degraded DNA: A comparison of three methods

Sex determination is of paramount importance in forensic anthropology. Numerous anthropological methods have been described, including visual assessments and various measurements of bones. Nevertheless, whatever the method used, the percentage of correct classification of a single bone usually varies between 80% and 95%, due to significant intra- and inter-population variations, and sometimes variations coming from secular trends. DNA is increasingly used in a forensic context. But forensic DNA extraction from bone raises several issues, because the samples are very often badly altered and/or in very small quantity. Nuclear DNA is difficult to get from degraded samples, according to low copy number, at least in comparison with mitochondrial DNA. In a forensic context (as in a paeleoanthropological context) DNA sex determination is usually complicated by the weak amount of DNA, the degraded nature of nucleic acids, the presence of enzymatic inhibitors in DNA extracts, the possible faint amplification of Y band and the risk of contamination during either excavation or manipulation of samples.The aim of this work was to compare three methods of DNA sex determination from bones: procedure #1 using a single PCR amplification, procedure #2 using a double PCR amplification, and procedure #3 adding bleaching for decontamination of the bone, instead of simply rubbing the bone. These processes were applied to samples of bones (49 samples coming from 39 individuals) that were in various states of post mortem alteration.The main results are the following. (i) No DNA could be extracted from three skulls (parietal bones, mastoid process), the compact bone of one rib, and the diaphysis of one femur; (ii) there was a contamination in three skulls; and (iii) the Y band did not appear in two male cases, with one of the three procedures (male tibia, procedure #2) and with procedures #2 and #3 (male femur).This study emphasises the main issue while working with altered bones: the impossibility to extract DNA in some cases, and, worth of all, the contamination of the sample or the faint amplification of Y band which leads to a wrong sex answer. Multiple and significant precautions have to be taken to avoid such difficulties.     

急性博落回中毒的实验病理学研究

目的 探讨博落回中毒作用的靶器官、靶组织和病变特点,以及博落回中毒机理。方法 应用博落回水煎液对大鼠经口急性染毒,行组织病理学和重要脏器电镜检查及血清生化检测。结果 博落回对大鼠的半数致死量(LD50)为19.5g/kg,染毒大鼠血清谷丙转氨酶(GPT)、肌酐(Cr)、尿素氮(BUN)、肌酸激酶(CK)及其同功酶(CK-MB)增高;心电图(EKG)表现异常;肝、脾、肾脏器系数减小。光镜下中毒大鼠心、脑、肝、肾等器官呈淤血、出血性改变;电镜下中毒大鼠心、脑、肝、肾细胞内线粒体、内质网、核膜等膜性结构破坏。结论 博落回毒作用的主要靶器官或靶组织是心、脑、肝、肾,病变特点为受累脏器淤血、出血性变;毒作用的主要机理是对线粒体、内质网和核膜等膜性结构的破坏。     

血管电损伤的实验研究

观察电击后血管病理改变的规律及特点 ,探讨其损伤的发病机制和诊断电击伤 (死 )的价值。采用Wistar大鼠复制血管电击伤模型 ,通过HE、GS、Weigert弹力纤维染色和α actin免疫组化染色 ,观察电击后不同节段血管的病理变化。结果表明 :(1)电击后血管的损伤范围和程度均较其周围大 ;(2 )电击部位的血管常发生全层凝固性坏死 ,血管周围骨骼肌、脂肪细胞广泛坏死 ,大腔隙形成 ;距电击部位越远损伤越轻 ,血管损伤局限于管壁中内层 ,而血管周围组织相对完好 ;(3)光镜下 ,血管内皮细胞不同程度坏死、脱落 ;平滑肌肌浆凝聚 ,呈嗜碱性 (异染 ) ,胞核深染 ,扭曲伸长呈栅栏状排列 ,细胞内外大量汽化空泡形成 ,肌层中弹力纤维减少 ,网状纤维相互融合增粗、断裂 ,呈串珠状改变。TEM下见血管内皮细胞、平滑肌细胞内线粒体、内质网不同程度肿胀与扩张 ,甚至空泡化改变 ,平滑肌肌丝溶解 ,形成低电子密度区 ;汽化空泡边界清楚 ,无膜性结构 ,内无细胞器 ;SEM下见血管壁及腔面粗糙大小不等的腔隙形成 ;(4 ) 2 2 0V组血管的病理改变与 10 0 0V组相同 ,只是损伤程度较前者轻。电击后血管的病理改变是电热效应与电场作用的共同结果 ,血管的病理改变可以作为一种确证电击死的重要依据     

Genetic markers in human bone: I. Deoxyribonucleic acid (DNA) analysis.

Deoxyribonucleic acid (DNA) was isolated from a number of spongy and compact human bone tissue specimens, and the yield was estimated on a "per milligram of starting tissue" basis. DNA was, in addition, isolated from a number of corresponding blood and bone tissue specimens. Spectrophotofluorometry and ethidium bromide visualization on minigels were used to estimate the quantity and degree of degradation of DNA. The DNA from several blood-bone pairs is shown to give concordant restriction fragment length polymorphism (RFLP) typing results by two different typing protocols with five different single-locus probes. DNA from several additional blood-bone pairs is shown to give concordant results for human leucocyte antigen (HLA)-DQ alpha phenotypes following polymerase chain reaction (PCR) amplification and hybridization to specific allele-specific oligonucleotide (ASO) probes, and for the variable numbers of tandem repeats (VNTR) length polymorphisms 3' to the human apolipoprotein B (APOB) gene following PCR amplification with specific primers and analysis of the products by electrophoresis and ethidium bromide visualization.     

电击伤组织的超微结构观察

目的 观察电流传导路径上器官、组织的超微结构变化。 方法 对 SD大鼠进行实验性低电压电击。结果 触电部位皮肤的上皮细胞胞浆内基质凝固、膜系统断碎;骨骼肌肌小节出现异常收缩带;心肌的变化也较明显,可见局灶性溶解坏死并出现异常收缩带;电流传导路径上外周血管、神经的改变以血管明显,血管内皮细胞胞浆中出现空泡,中膜平滑肌可见溶解坏死,神经纤维仅见部分髓鞘松解。 结论 电击伤组织的超微结构变化可作为诊断电击死组织学变化的补充,为解决无明显皮肤电损伤的电击死案件提供形态学指标。同时也探讨了电击伤超微结构变化的形成机制。     

Chronic Alcohol Abuse Leads to Low Bone Mass with No General Loss of Bone Structure or Bone Mechanical Strength,

Chronic alcohol abuse (CAA) has deleterious effects on skeletal health. This study examined the impact of CAA on bone with regard to bone density, structure, and strength. Bone specimens from 42 individuals with CAA and 42 individuals without alcohol abuse were obtained at autopsy. Dual‐energy X‐ray absorptiometry (DEXA), compression testing, ashing, and bone histomorphometry were performed. Individuals with CAA had significantly lower bone mineral density (BMD) in the femoral neck and significantly lower bone volume demonstrated by thinner trabeculae, decreased extent of osteoid surfaces, and lower mean wall thickness of trabecular osteons compared to individuals without alcohol abuse. No significant difference was found for bone strength and structure. Conclusion: CAA leads to low bone mass due to a decrease in bone formation but with no destruction of bone architecture nor a decrease in bone strength. It is questionable whether this per se increases fracture risk.     

TW2、TW3法腕骨骨龄评价方法的诊断性试验

目的比较TW2法英国人标准、TW2法中国人南方标准和TW3法测定腕骨骨龄对特发性性早熟(IPP)女孩的诊断价值。方法女性IPP患儿55例;对照组83例。回顾性分析初诊左腕X线片,用TW2法、TW2CHC中国人南方标准和TW3法盲法分期,分别测得其腕骨骨龄及其百分位,并分为5个连续性的工作点,即:>P97、>P90、>P75、>P50和、≤P50。结果敏感度(Sen)和特异度(Spe)都比较高的有以下3个工作点:TW2CH“>P90”点,TW3“>P90”点,TW2“>P90”点。结论TW系统3种腕骨骨龄对于IPP女孩都有诊断价值,TW3法和TW2法中国人南方标准诊断价值比较高,TW2法英国人标准的诊断价值为中等。     

Human Fatality Due to Thallium Poisoning: Autopsy,Microscopy, and Mass Spectrometry Assays,

Thallium has been responsible for many intoxications since its discovery; however, toxicological profiles for thallium in human fatalities have not been updated recently. Autopsy, microscopic investigations, and toxicological analyses were performed on a married couple who died from thallium sulfate intended homicidal poisoning. The distribution of thallium was established by inductively coupled plasma mass spectrometry with hair samples showing the highest thallium concentration. Electron microscopy revealed a dystrophic condition of hair with disorganized cuticle and atrophy of the hair bulb. Thallium interacts with cells at different levels, with prominent ultrastructural injuries in the mitochondria and endoplasmic reticulum, and high concentration of electron dense granules observed in the cytoplasm and mitochondria of several organs. Alopecia, toxic encephalopathy, and peripheral neuropathy were diagnosed in the victims and suggested to be crucial implications for thallium poisoning. The analytical procedures used in this case are of considerable forensic importance in the diagnosis of thallium poisoning.     

不同时程吗啡依赖大鼠依赖相关脑区光镜和电镜观察

目的探讨吗啡依赖中枢神经系统毒理病理不同时程的变化。方法背部皮下递增注射吗啡建立吗啡依赖大鼠模型,光镜和透射电镜下观察不同时程大鼠吗啡依赖相关脑区,即蓝斑核、中脑导水管周围灰质、黑质、豆状核、杏仁核、海马的病理组织学变化,并对突触进行计数,比较与空白对照组的差异。结果吗啡依赖大鼠6个依赖相关脑区均出现神经细胞固缩或肿胀,神经纤维肿胀,线粒体肿胀、畸形,内质网扩张,多聚核糖体解聚,突触数量增多;胶质细胞、脑膜下淋巴细胞增多,随依赖时间的延长而更加显著,并可见胶质结节形成。吗啡依赖8周组、4周组, 大鼠突触数量较空白对照组增多,其差异具有非常显著性意义(P<0．01);吗啡依赖8周组大鼠突触数量的增多,与吗啡依赖1周组、2周组大鼠比较,其差异具有非常显著性意义(P<0．01)。结论吗啡依赖大鼠依赖相关脑区神经细胞呈缺血缺氧性及退行性改变,突触数量增多,并随吗啡依赖时限的延长而明显。     

Postmortem evaluation of barbiturate poisonings

The evaluation of barbiturate intoxication as the cause of death is often difficult when the concentration in body fluids and organs is not extremely high. The problem arises because of the great capacity of barbiturates to produce tolerance after chronic use, a property that is often unknown. Therefore, the most abused barbiturates were studied to assess whether chronic intake causes morphological liver changes or not. It was found that the chronic abuse of drugs containing seco-, cyclo-, brallo-, and/or pentobarbital produces hypertrophy of the smooth endoplasmic reticulum of hepatocytes corresponding to that of phenobarbital. Neither acute barbiturate overdose (without a history of abuse) nor chronic abuse of opiates causes similar liver changes. In conclusion, barbiturate tolerance can be evaluated postmortem by light microscopic examination of the liver.     

Utility of immunoassay in drug screening in skeletal tissues: sampling considerations in detection of ketamine exposure in femoral bone and bone marrow following acute administration using ELISA

Detection of ketamine exposure in skeletal tissues by automated enzyme-linked immunosorbent assay (ELISA) and gas chromatography with electron capture detection (GC-ECD) is described. Rats (n = 18) received 0, 15, 30, or 75 mg/kg ketamine hydrochloride acutely (i.p.), and were euthanized within 15 min or 1 h. Ketamine was extracted from ground femoral bone by methanolic incubation followed by liquid-liquid extraction (LLE), while marrow was homogenized in alkaline solution, and then underwent LLE. Extracts were analyzed by ELISA, and subsequently by GC-ECD following derivatization with trifluoroacetic acid anhydride. The effect of tissue type (i.e., diaphyseal bone vs. epiphyseal bone vs. bone marrow) on the immunoassay response was examined through determination of binary classification test sensitivity (S) and measurement of the relative decrease in absorbance (%DA, drug-positive tissues vs. drug-free controls) in each tissue type. The %DA varied significantly between different tissues examined under a given dose condition, and generally decreased in the order marrow > epiphyseal bone > diaphyseal bone, at all dose levels examined. Measured S values for marrow, epiphyseal bone, and diaphyseal bone were 100%, 77%, and 23%, respectively (75 mg/kg dose). These results suggest that the type of skeletal tissue sampled and position sampled within a given bone (diaphyses vs. epiphyses) are important parameters in drug screening of skeletal tissues.