骨骼肌circRNA在死亡时间推断中的初步筛选及验证研究

目的通过构建骨骼肌中环状RNA(circular RNA,circRNA)表达谱,筛选出人体骨骼肌中与死亡时间具有时序性变化的circRNAs,并对筛选出的circRNAs进行验证,进而探讨其相对表达量与死亡时间的相关性。方法芯片实验选取2例外界温度、湿度相对一致,已知死亡时间在24 h内的个体,分别于尸检即刻(12 h以内)、3、5、7、10 d各取50 g组织进行总RNA提取,并进行逆转录、探针标记、基因芯片检测等实验,初步得到circRNA表达谱;对筛选出的circRNAs在后续收集的18例检材中进行验证,对所得数据进行统计分析。结果骨骼肌组织中共检测出13 156个circRNAs,但在10 d内整体呈下降趋势的circRNAs只有hsa_circ₀001727、hsa_circ₀001136、hsa_circ₀001871、hsa_circ₀005251、hsa_circ₀     

Anthropological analyses of 30 insertion/deletion autosomal markers in five major ethnic groups of Pakistan

We investigated the forensic efficacy of the 30 insertion/deletion (Indel) markers included in the Qiagen Investigator® DIPplex kit in 529 Pakistani individuals from five major subpopulations in Pakistan (Punjabi, Pashtun, Sindhi, Saraiki, and Baloch). In the Sindhi population, the distribution of HLD81 and HLD97 alleles deviated from Hardy-Weinberg equilibrium after Bonferroni correction. The combined match probability ranged from 2.0E-12 (Pashtun and Baloch) to 1.0E-12 (Sindhi), and the mean paternity exclusion power varied from 0.995 (Punjabi, Sindhi, and Saraiki) to 0.996 (Pashtun and Baloch). The high combined power of discrimination (0.999 999 999 999 97) and low combined match probability (1.7E-12) for all subpopulations studied support the utility of the 30 Indel markers for forensic identification in the studied subpopulations. The allele frequencies of the Indel markers in the Pakistani subpopulations were compared with those from 18 other populations. The results show that the populations clustered according to geography. The subpopulations investigated in this work showed a close genetic relationship with others from Pakistan, as well as with South Central Asian and Middle Eastern populations. The results suggest that the Investigator^® DIPplex kit can be useful as a supplementary tool for human identification in the five Pakistani subpopulations investigated in this study.Supplemental data for this article is available online at https://doi.org/10.1080/20961790.2021.1933366 .     

Age estimation of individuals aged 5–23 years based on dental development of the Indonesian population

Dental development can be used to estimate age for forensic purposes. However, most of the currently available methods are less reliable for the Indonesian population due to population variability. This study presents a new method and evaluates other methods that utilize dental development to estimate the age of Indonesian people. Panoramic radiographs of 304 young Indonesian people aged 5–23 years old were analysed for deciduous tooth root resorption, permanent tooth calcification, and eruption. The extent of tooth root resorption was determined based on AlQahtani’s modified Moorrees et al. method. Tooth calcification was classified based on a modified Demirjian et al. method. Tooth eruption was evaluated based on AlQahtani’s modified Bengston system. The sequence of tooth root resorption, and permanent tooth calcification and eruption were grouped into 19 age categories (from 5–23 years old) in an atlas. The differences between males and females, between maxillary and mandibular teeth, and between right and left teeth were also analysed. There were minimal significant differences of tooth development between males and females, and between the right and left teeth (P > 0.05), while the maxillary and mandibular dental development was significantly different (P < 0.05). The newly developed atlas showed the development of the right side of maxillary and mandibular tooth of combined sex of Indonesian population. Another 34 panoramic radiographs of known-age and sex individuals from Indonesia were assessed using the newly developed Atlas of Dental Development in the Indonesian Population, Ubelaker’s Dental Development Chart, The London Atlas of Human Tooth Development and Eruption by AlQahtani, and the Age Estimation Guide-Modern Australia population by Blenkin-Taylor. Accuracy was assessed by comparing estimated age to actual chronological age using the Bland-Altmand test. Results show that the smallest range of error was found in the Atlas of Dental Development in the Indonesian Population (−0.969 to 1.210 years), followed by The London Atlas of Human Tooth Development and Eruption by AlQahtani (−2.013 to 1.990 years), the Age Estimation Guide-Modern Australia population by Blenkin-Taylor (−2.495 to 2.598 years), and the Dental Development Chart by Ubelaker (−2.960 to 3.289 years). These findings show that the Atlas of Dental Development constructed in this study performs better than the other three methods and presents greater accuracy of age estimation in the Indonesian population.

Key points

• Dental development such as deciduous tooth root resorption, permanent tooth calcification, and tooth eruption can be used to estimate age for forensic purposes.
• The development of the teeth are influenced by genetic, ethnicity, and sex, therefore an age estimation method must be constructed based on the same population.
• There were minimal significant differences in tooth development between male and female, and between right and left teeth, but there was significant difference between maxillary and mandibular teeth.
• The Atlas of Dental Development in the Indonesian Population constructed in this study allowed more accurate age estimation of the Indonesian sample than the other methods tested.

Supplemental data for this article are available online at https://doi.org/10.1080/20961790.2021.1886648.     

Analysis of sensitivity and specificity: precise recognition of neutrophils during regeneration of contused skeletal muscle in rats

In this report, we applied the TissueFAXS 200 digital pathological analysis system to rapidly and accurately identify neutrophils during regeneration of contused skeletal muscle, and to provide information for follow-up studies on neutrophils to estimate wound age. Rat injury model was established, and skeletal muscle samples were obtained from the control group and contusion groups at 1, 1.5, 2, 3, 4, and 6 h, as well as at 1, 3, 5, and 15 d post-injury (n = 5 per group). The expression of nuclei and neutrophils was detected by hematoxylin and eosin (HE) staining and immunohistochemical (IHC) staining. A total of 20 injury site areas of 0.25 mm² (0.5 mm × 0.5 mm) were then randomly selected at all time points. A TissueFAXS 200 digital pathological analysis system was used to identify the positive and negative numbers. Knowledge of five professional medical workers were considered the gold standard to measure the false positive rate (FPR), false negative rate (FNR), sensitivity, specificity, and area under the curve (AUC) of receiver operating characteristic (ROC) curves. As a result, with a staining area of neutrophils from 8 µm² to 15 µm², the FPR was 4.28%–12.14%, the FNR was 12.42%–64.08%, the sensitivity was 35.92%–87.58%, the specificity was 87.86%–95.72%, the Youden index was 0.316–0.754, the accuracy was 82.80%–88.30%, and the AUC was 0.771–0.826. The AUC was largest when the cut-off value of the staining area was 12 µm². Our results show that this software-based method is more accurate than the human eye in evaluating neutrophil infiltration. Based on the sensitivity and specificity, neutrophils can be accurately identified during regeneration of contused skeletal muscle. The TissueFAXS 200 digital pathological analysis system can also be used to optimize conditions for different cell types under various injury conditions to determine the optimal cut-off value of the staining area and provide optimal conditions for further study. Furthermore, it will provide evidence for forensic pathology cases.     

犬细小病毒JL-(18)1/Beagle株的分离鉴定及遗传进化分析

为研究犬细小病毒(CPV)流行株遗传变异情况,本研究从吉林省某比格犬养殖场采集疑似CPV感染致死犬的肠道组织,将病料接种至F81细胞进行病毒分离,并通过电镜形态学、PCR、血凝试验及动物回归试验进行鉴定.结果显示,所分离的这株病毒在F81细胞中产生了典型CPV细胞病变(CPE);在电镜下可见病毒呈无囊膜、直径为20 n...     

指纹鉴定标准及鉴定结论概率化研究

指纹鉴定标准是指纹鉴定理论的核心内容之一.指纹鉴定理论和实践中形成的最低特征数量标准、质量—数量标准、面积—质量量化标准、拓扑学方法和标准、形态学方法和标准等均有其局限性.运用统计学理论和贝叶斯定理建立指纹鉴定结论概率化方法和评价标准,不仅在理论和实践上具有必要性和可行性,而且也可以避免指纹鉴定标准之争,使指纹鉴定从经验走向科学.通过统计每枚指纹平均细节特征数量和每类特征的出现率,可以计算出两枚指纹匹配的概率值,进而对概率结论的可靠性作出分析评断.     

无骨折脱位型颈脊髓损伤法医学鉴定25例分析

目的 探讨无骨折脱位型颈脊髓损伤案例中外伤与损害后果因果关系法医学鉴定的要点.方法 收集25例无骨折脱位型颈脊髓损伤的法医学鉴定案例资料,总结被鉴定人的年龄、致伤方式、影像学检查结果、自身颈椎退变情况、临床表现、主要诊疗情况、预后情况等,按照损伤与损害后果相关程度分为直接原因、主要原因、同等原因、次要原因、无原因进行伤...     

Hair as a specimen to document tetramethylene disulfotetramine exposure

Tetramethylene disulfotetramine (tetramine) is a rodenticide that has been banned for many years in China. Since 2005, inhabitants of a village in the Henan Province have been suffering from grand mal seizures. To investigate the possibility of tetramine as the cause, we developed a method to determine tetramine in human hair. Sample preparation involved external decontamination, frozen pulverization, and ultrasonication in 2 mL ethyl acetate in the presence of cocaine-d3 as an internal standard. The method exhibited good linearity; calibration curve was linear over a range of 0.1-20 ng/mg hair. The limit of detection for the assay was 0.05 ng/mg hair. Except for one subject (No. 4), all head and pubic hair samples were positive for tetramine. The concentrations of tetramine in pubic hair were significantly higher than those in the same subjects' head hair samples. Because of a long retention in body, segmental head hair analysis cannot provide an accurate exposure history of tetramine in the body.