丛状骨和骨单位带的种属特征及法医学应用价值

目的 探讨丛状骨和骨单位带的种属特征及其法医学应用价值。方法 人及猪、牛、羊、狗长骨中段横切磨片35张，在光学显微镜下观察骨组织中丛状骨和骨单位带的形态学特征。结果 丛状骨和骨单位带在猪、牛、羊、狗的密质骨中广泛存在，人骨组织中几乎不见；同种动物不同个体之间骨组织学结构也存在差异。结论 观察丛状骨和骨单位带是根据人骨鉴别种属的重要排除指标。     

烧骨残片种属鉴定的研究

为了找出常见动物骨骼焚烧残片的种属鉴定方法,笔者对猪、羊,股骨及肱骨的断端,猪、羊、牛颅骨的残片(额顶部),焚烧后进行了观察及测量,并与人类股骨、肱骨及焚烧的颅骨残片进行了比较.结果发现,长骨滋养孔的位置,骨干骨壁厚度,以及颅骨厚度,颅缝形态及内板表面特征等方面,是鉴定骨骼残片种属的有价值的特征.根据对常见动物长骨及颅骨残片的现察及研究,提出了骨骼残片种属鉴定的方法.     

人和动物SON基因3’非编码区的PCR测序分析

目的 研究人和猕猴、阿拉伯狒狒、猪、牛、羊、马、驴、骡、狗、猫、兔、大白鼠、小白鼠、豚鼠等14种哺乳类动物SON基因3’非编码区(3’UTR)的种属特异性碱基序列差异。方法 对人和上述14种哺乳类的SON基因3’非编码区中的部分种属特异性区域进行PCR测序分析。结果 人有2条扩增片段和14种哺乳类动物各有1条扩增片段的碱基序列;人无关个体和同一种属不同个体动物DNA扩增片段的碱基序列相同,人、猕猴、阿拉伯狒狒、猪、牛、羊、马、驴、骡、狗、猫、兔、大白鼠、小白鼠、豚鼠DNA的SON基因3’UTR扩增片段之间存在碱基序列差异。结论 SON基因3’UTR是一个具有良好种属特异性的遗传标记,采用SON基因3’UTR扩增测序技术可以对人和上述14种哺乳类动物进行准确的种属鉴定。     

PCR扩增SON基因3’非编码区进行种属鉴定

根据人 DNA的 SON基因碱基序列选择性设计一对特异性引物 ,并对人和猕猴、阿拉伯狒狒、猪、牛、羊、马、驴、骡、狗、猫、兔、大白鼠、小白鼠、豚鼠等 14种哺乳类动物染色体 DNA的 SON基因 3’非编码区中的种属特异性区域进行 PCR扩增 ,扩增产物经 SSCP分离银染色技术检测 ,观察到人和 14种哺乳类动物的 SSCP图谱有明显不同。本方法可以对人和上述 14种哺乳类动物进行种属鉴定 ,适合于法医学种属鉴定的应用     

扩增Amelogenin基因用于生物检材种属鉴定

目的扩增Amelogenin基因测定血痕或组织的种属,以确定在法医学检验中的应用价值。方法收集猪、羊、马等10几种常见动物的血痕或肌肉组织,应用PCR扩增Amelogenin基因,PAGE电泳,银染后观察结果。结果哺乳动物猪、牛、狗、羊、马、驴动物血痕扩增产物为1条带,片段长度102bp。猕猴检见106bp和112bp两条带,与人血痕没有区别。兔、猫、鼠血痕未检出特异性片段。其它常见物种鳝鱼、青蛙、鸭、鹅、鸽、鹌鹑、麻雀等均未见扩增产物。结论扩增Amelogenin基因进行种属鉴定,方法简单,灵敏度高,可应用于法医检案。     

人和猪、羊、牛肌组织形态的比较及法医学意义

目的观察不同种属肌组织形态的特征指标,建立种属鉴别的判别方程,尝试建立一种种属鉴别新方法。方法分别提取20名成年死者三个部位(肱三头肌、股二头肌和竖脊肌)的肌组织和达到出栏年龄的猪、羊、牛各20头三个部位(臂三头肌、股二头肌和背最长肌)的肌组织,制成石蜡切片。测量人及猪、羊、牛肌组织的11项观察指标,并应用统计学方法进行分析,建立种属判别方程。结果筛选出4项观察指标建立人与猪、羊、牛之间种属鉴别的判别方程,对人肌组织的判别准确率可达90%,对猪、羊、牛肌组织的判别准确率分别可达80%、100%和80%。结论形态学方法为人与猪、羊、牛之间肌组织的种属鉴别提供了一种新的方法,可为动物种属鉴别提供参考依据。     

骨骼种属鉴定的组织学研究

目的 探求骨骼残渣的种属鉴定的组织学方法。方法 对人体不同部位骨骼,及常见动物骨骼的 77张骨组织学切片,在光镜下进行了骨组织学形态学研究。结果 不同动物骨组织学的形态学特征存在明显差异,其主要差别有：内、外环骨板的形态和相对厚度;哈氏系统之间的骨板的形态和哈氏系统的大小;哈氏环层骨板的数量及骨陷窝的数量等。结论 骨组织学的特征,不仅可以区分人与动物骨骼的残渣,也可以对不同动物的骨骼残渣进行区分。     

扩增TP53内含子8用于生物检材的种属鉴定

目的扩增常见动物TP53内含子8片段,确定其在法医学生物检材种属鉴定中的应用价值。方法收集标本为包括人在内的15种常见动物的血痕或肌肉组织,提取DNA后定量,应用PCR扩增TP53内含子8,PAGE电泳,银染后观察结果。结果人和猕猴都扩增出一条长度为460bp的片段,鳝鱼、鲢鱼、青蛙、鸭、兔、猫、小白鼠、豚鼠、猪、牛、羊虽有扩增产物,但不在分型区内,鸡、狗未见扩增产物。结论扩增TP53内含子8进行种属鉴定,方法简单,灵敏度较高。     

根据股骨组织形态学变化进行骨骼种属鉴别

目的通过对成年人、成体黄牛、马、家猪、山羊、绵羊、狗、猫、长毛兔、鹅、鸭、鸡共12个种属动物股骨组织形态学结构特征的研究,建立一个有效的各类动物间种属鉴别的方法。方法在征得家属同意后,在尸检中取成人(20岁以上)右股骨中段约4cm,同时,收集黄牛、马、家猪、山羊、绵羊、狗、猫、长毛兔、鹅、鸭、鸡共11种常见动物右腿股骨,取中段约4cm,脱钙后制作成骨组织切片,在光学显微镜下观察,将显微镜下的图像录入电脑,选取24个指标进行分析。结果人与其他动物、以及各被检动物之间在骨单位数量等13个指标上具有显著差异,用这些指标建立种属判别数学模型,结果这些动物之间的判别率也可达89.4%。结论股骨中段骨密质的组织学结构具有明显的种属特征,且骨单位形态和数量呈现明显的生物进化趋势。根据这些特征可以有效地进行种属鉴别。     

线粒体16SrRNA和Cytb基因复合扩增进行种属鉴定

目的 建立一种用于种属鉴定的线粒体DNA16SrRNA基因和细胞色素b基因荧光标记复合扩增检测体系。方法 利用引物设计软件Primer 5．0对mtDNA序列的16SrRNA基因和细胞色素b基因各设计一对引物,建立复合扩增体系,分别扩增人和牛、猪、狗、鸡、草鱼5种常见动物,用310遗传分析仪对产物进行分析。结果 人和5种动物DNA扩增产物均出现两个峰。Cytb通用引物的扩增产物为人与动物的共有峰,为358bp;16SrRNA基因的扩增产物为人与动物间存在位置差异的特异峰,位于231～256bp之间。结论 该复合扩增体系可以明确区分人和5种动物样本,可用于种属鉴定。     

Differentiating human bone from animal bone: a review of histological methods

This review brings together a complex and extensive literature to address the question of whether it is possible to distinguish human from nonhuman bone using the histological appearance of cortical bone. The mammalian species included are rat, hare, badger, racoon dog, cat, dog, pig, cow, goat, sheep, deer, horse, water buffalo, bear, nonhuman primates, and human and are therefore not exhaustive, but cover those mammals that may contribute to a North American or Eurasian forensic assemblage. The review has demonstrated that differentiation of human from certain nonhuman species is possible, including small mammals exhibiting Haversian bone tissue and large mammals exhibiting plexiform bone tissue. Pig, cow, goat, sheep, horse, and water buffalo exhibit both plexiform and Haversian bone tissue and where only Haversian bone tissue exists in bone fragments, differentiation of these species from humans is not possible. Other primate Haversian bone tissue is also not distinguishable from humans. Where differentiation using Haversian bone tissue is undertaken, both the general microstructural appearance and measurements of histological structures should be applied. Haversian system diameter and Haversian canal diameter are the most optimal and diagnostic measurements to use. Haversian system density may be usefully applied to provide an upper and lower limit for humans.     

Histological Determination of the Human Origin of Bone Fragments

Abstract:  A frequently encountered task in the forensic scenario is verification of the human origin of severely degraded fragments of bone. In these cases histological methods which consider osteon size and morphology can prove to be useful. The authors in the present study verify the applicability of published algorithms to flat and subadult bones from human, dog, cat, cow, rabbit, sheep, pig, chicken, quail, and turkey samples. Metric analysis was performed on 2031 Haversian canals. Analyses carried out on human samples confirmed a success rate of around 70% on long adult bones; however the percentage of wrong answers was particularly high in the case of newborns and older subadults as well as on flat bones in general. Results therefore suggest that such regression equations should be limited only to bone fragments from long adult bones.     

Estimation of age from the femur of Japanese cadavers

The purpose of this study is to estimate the age of cadavers by histomorphometry of the femur. Seventy-two Japanese males ranged from 43 days to 92 years old and 26 females ranged from 2 to 88 years old were used. The thickness of sections was adjusted at 50 to 70 microns by grinding with sand paper. The sections were not decalcificated. They were stained with Villanueva's bone staining powder and with thionin dye. Microradiographs of the sections were obtained by the soft X-ray apparatus. The area, maximum and minimum diameter, and perimeter of the perfect osteon and Haversian canal were measured. In addition, the type II osteon number, osteon fragment number, and area of triangle were also determined. All these parameters were examined by an image analyzer. The parameters of the osteon showed high correlation coefficient with age (magnitude of r > 0.77), while those of the Haversian canal were low (magnitude of r < 0.11). All parameters were subjected to multiple regression analysis for producing a multiple regression equation of age estimation. For the stepwise selecting method, the perimeter of osteon, maximum length of the Haversian canal and osteon fragment number were selected for the equation. Their multiple r2 and standard error of estimation were 0.8874 and 6.39, respectively. For the forward selection method, in addition to the above items, three parameters, the maximum length of Haversian canal, triangle area, osteon fragment number were selected. Their multiple r2 and standard error of estimation were 0.9484 and 4.884, respectively. Bone staining was useful to clarify the demarcation between osteon and fragment, leading to an increase in the accuracy of age estimation. However, the entire range from birth to 90 years was difficult to cover for precise age estimation.     

人与动物骨组织特征的比较研究

目的研究区别骨骼残片是否为人类骨骼的方法。方法提取人体骨骼及鱼类、两栖类、爬行类、鸟类、哺乳类、灵长类等动物的骨骼标本制做骨骼组织学片,在显微镜下观察,将显微镜下的图像录入电脑进行分析。结果内、外环骨板,骨单位,骨细胞的显微镜下的结构,人类与动物存在明显差别。结论根据骨骼的组织学特征可以区别人类与动物的骨骼。     

Microscopic Pattern of Bone Fractures as an Indicator of Blast Trauma: A Pilot Study

The assessment of fractures is a key issue in forensic anthropology; however, very few studies deal with the features of fractures due to explosion in comparison with other traumatic injuries. This study focuses on fractures resulting from blast trauma and two types of blunt force trauma (manual compression and running over), applied to corpses of pigs; 163 osteons were examined within forty fractures by the transmission light microscopy. Blast lesions showed a higher percentage of fracture lines through the Haversian canal, whereas in other types of trauma, the fractures went across the inner lamellae. Significant differences between samples hit by blast energy and those runover or manually compressed were observed (p < 0.05). The frequency of pattern A is significantly higher in exploded bones than in runover and compressed. Microscopic analysis of the fracture line may provide information about the type of trauma, especially for what concerns blast trauma.     

Differentiating fragmented human and nonhuman long bone using osteon circularity

Distinguishing between human and nonhuman bone is important in forensic anthropology and archeology when remains are fragmentary and DNA cannot be obtained. Histological examination of bone is affordable and practical in such situations. This study suggests using osteon circularity to distinguish human bone fragments and hypothesizes that osteons will more closely resemble a perfect circle in nonhumans than in humans. Standard histological methods were used, and circularity was determined using an image analysis program, where circularity was controlled for by Haversian canal measurements. Homogeneity was first tested for multiple variables within human and nonhuman samples. No significant differences were found between human sexes (p = 0.657) or among nonhuman species (p = 0.553). Significant differences were found among intraskeletal elements of both humans (p = 0.016) and nonhumans (p = 0.013) and between pooled samples of humans and nonhumans (p < 0.001). Results of this study indicate that osteon circularity can be used to distinguish between fragmented human and nonhuman long bone.     

Microscopic Age Estimation from the Anterior Cortex of the Femur in Korean Adults*

Abstract: The purpose of this study was to develop age‐predicting equations from the anterior cortex of the femur of Korean adults. Seventy‐two femoral samples (44 male and 28 female) were obtained from Korean cadavers and used to develop the equations. The thin sections (<100‐μm thick) were prepared by manual grinding; the sections were not decalcified and were stained with Villanueva bone stain reagent. Analysis of covariance showed no significant differences in age‐adjusted histomorphological variables between sexes. In stepwise regression analysis, osteon population density, average osteon area, and the most anterior cortical width were selected for an age‐predicting equation which produced a high regression correlation (R² = 0.789). The average Haversian canal area was not significantly related to age for any specimen.     

Differences among species in compact bone tissue microstructure of mammalian skeleton: use of a discriminant function analysis for species identification

In order to develop an identification key for distinguishing between several mammalian species, bone structure of their compact bone tissue was analyzed using qualitative and quantitative characteristics. Ninety femora of adult male humans, pigs, cows, sheep, rabbits, and rats were studied. The average area, perimeter, minimum, and maximum diameter of 1863 Haversian canals and 1863 secondary osteons were measured using a digital image device. The observed data were first used to evaluate inter- and intraspecies diversity. After that, we applied a discriminant function analysis for differentiation of the species by these variables. Classification functions for investigated species give cross-validated correct classification rates for 76.17% of cases. This percentage value can be increased by integrating conclusions from the qualitative analysis.