Towards molecular autopsies: Development of a FFPE tissue DNA extraction workflow

Sudden unexpected death (SUD) is a devastating event and forms a substantial proportion of the cases investigated at forensic mortuaries each year. Despite post-mortem investigations, the cause of death may remain undetermined. There is potential for these unresolved cases to benefit from retrospective molecular autopsies for investigation into genetic mutations which may have contributed towards death. Often, formalin fixed paraffin embedded tissues (FFPET) are the only archival sources of DNA available for retrospective analyses. However, extracting usable DNA from FFPET is challenging as current methods yield poor quality and quantity DNA. Thus, this study aimed to optimise DNA recovery from FFPET by investigating several variables within the DNA extraction workflow, including the selection of tissue type, number and thickness of tissue sections, deparaffinisation method, and DNA extraction kit. The quantity and quality of DNA recovered were assessed using spectrophotometry, real time PCR, digital capillary electrophoresis and DNA profiling. This study was the first to implement a nuclei quantification using microscopy to guide the selection of the best tissue type to use for DNA analysis. The use of a greater number of thinner tissue sections (100 sections, each 1 μm) significantly improved DNA concentration, purity and fragment length. Additionally, the combination of Deparaffinization Solution with the QIAamp® DNA FFPE Tissue Kit proved most favourable with a median DNA yield of 320 ng and 55% of DNA fragments greater than 400 bp. Isolated DNA was of single source, indicating no contamination in the workflow, and FFPET blocks that were stored for up to 3.5 years did not significantly affect DNA degradation (p = 0.1764). These results are especially informative for designing library preparation and sequencing workflows for determining cause of death in unresolved SUD cases.     

Position Effect on Facial Soft Tissue Depths: A Sonographic Investigation

The head is positioned erect for an approximation; yet most facial soft tissue depths (FSTD) used are measured from supine subjects. Depth difference might be significant, but there is a paucity of data to verify. This study compared erect and supine values for 17 landmarks from 30 healthy New Zealand (European population affinity) women (18–30 or 40–55 years) in erect then supine positions. Height, weight, and sonographic FSTD data, totaling 1020 measurements, were obtained. Three midline and seven averaged bilateral values were compared using ANOVA, p values, and Pearson's correlations. Correlative strength of age and body mass index, BMI (kg/m²), was determined by values. Results showed averaged erect and supine differences were significant for four of ten FSTDs. Between individuals, difference was various and not unidirectional. In conclusion, depth differences were observed but not all significant or unidirectional, BMI significantly influenced nine FSTD values, but age group did not.     

Vertical Lip Position and Thickness in Facial Reconstruction: A Validation of Commonly Used Methods for Predicting the Position and Size of Lips

This study examined several methods used to estimate oral fissure position, lip margin position, and lip thickness recommended by Angel, George, Lebedinskaya, Taylor, Wilkinson et al., Balueva and Veselovskaya. A sample of 86 lateral head cephalograms of adult subjects from central Europe were measured and the actual and predicted dimensions were compared. The best estimation for oral fissure position was “opposite the lower ¾ mark of maxillary incisors” (error of 1.3 mm). Upper lip margin was predicted best by “upper ¼ mark of maxillary incisors” (error of 1.7 mm), and lower lip margin by “cementum‐enamel junction of mandibular incisors” (error of 2.3 mm). The regression equations of Wilkinson et al. displayed least error (1.3 mm and 1.8 mm, respectively) for upper and lower lip thickness, and method of George (error of 3.4 mm) for total lip thickness.     

mRNA在体液斑迹鉴定与组织来源推断中的应用

体液斑迹鉴定及其组织来源推断一直是法医学研究的重要方向之一,其传统的检验方法存在诸如假阳性率高、检材易破坏等缺陷,亟需更高效的确证实验。高度分化的体细胞表达特异的m RNA分子,可推断其来源:外周血、月经血、精液、唾液、阴道分泌物、接触斑等,且灵敏度、特异度及保存时间相对较为理想,是未来用于体液斑迹鉴别的理想遗传标记。本文对m RNA在体液斑迹鉴定和组织来源推断方面的应用及其前景进行综述。     

论小产权房与我国土地的利益分配

小产权房是一个全国热议的话题,针对小产权房问题各家学者观点纷呈。与其说小产权问题首先涉及法律问题,不如说首先涉及利益分配的问题,处理小产权房问题,也就是对整个社会资源的再分配问题。在小产权房带来的这一场变化中,受益最大的是地方政府和一些房地产开发商。未来土地制度的设计必须充分考虑我国的国情,因此,必须结合我国人口资源环境长远发展道路,思考"小产权房"问题。     

广泛软组织挫伤--挤压综合征

本文通过回顾国内外医学文献,对广泛性软组织挫伤--挤压综合征的研究现状和取得的成就进行了介绍,并对其今后的发展趋势进行了探讨.     

推进长吉图开发开放先导区建设的政策建议——基于地区间产业关联研究的结论

推进长吉图开发开放先导区的建设对于培育吉林省新的增长极、提高东北地区开发开放水平意义重大。利用地区间/多区域投入产出模型,对东北地区和东北亚区域产业关联进行实证分析,发现东北地区在国际产业关联、区际产业关联、省际产业关联等三个方面的程度和效能都比较低,导致产业发展和产业结构调整严重受限。今后,吉林省需要积极培育和发展主导产业(群)、通过合理配套相关产业政策、加强基础设施建设,重点发挥长吉图开发开放先导区在东北地区"飞铃"型产业关联的中枢作用,带动东北地区全面发展。     

Rethinking ‘Crowding Out’ and the Return of ‘Private Affluence and Public Squalor’

This article traces the history of ‘crowding out’, and its use as a justification for austerity and state deflation from its origins in the 1920s to its latest post-2010 incarnation. It examines why governments have kept turning to austerity and continue to justify it on the grounds that public sector activity crowds out more productive private activity, despite the accumulated evidence that this traditional pro-market formulation has failed to deliver its stated goals. It examines three other embedded forms of crowding out that have been highly damaging—leading to weakened social resilience and more fragile economies—but which have been ignored by both governments and mainstream political economists.     

Alprazolam and Zolpidem in Skeletal Tissue of Decomposed Body Confirms Exposure

In several medico‐legal cases, bone samples analysis may provide the only source of toxicological information. This case study reports the analysis of a human bone specimen, belonging to a 46‐year‐old man, found 3 months after his death due to cervical–thoracic injuries in a motorcycle accident. Bone specimen was the only available material for toxicological analysis, among few skull hair and rotten skin. Analysis was performed by a newly developed and validated ultra‐high‐pressure liquid chromatography–mass spectrometry/mass spectrometry (UHPLC‐MS/MS) method, following simple and efficient sample pretreatment. The results were in accordance with the man's medical record: Alprazolam and zolpidem were found at 2.2 and 5.4 ng/g of bone, respectively. Both these drugs were prescribed to the deceased.