用PCR—测序法对人类线粒体DNA多态区的研究

用PCR方法，对线粒体DNA多态区15997至三6401区域进行扩增，产物DNA经纯化处理局直接进行自动化序列测定，得到了准确的序列结果。将得到的中国人mtDNA多态区序列与国外已报道的相应序列进行对比，证实我们所测序列与白种人相应序列之间存在碱基差异。     

二代测序技术在法医学中的应用进展

近几年二代测序(second generation sequencing,SGS)技术的快速发展,使其在通量增大、读长增加的同时测序成本大幅降低,给生物学领域带来了新的突破,也使法医遗传学进入了一个新的发展阶段。本文从测序技术在法医遗传学的应用历程展开,回顾了测序技术对法医学遗传标记检测的重要性。以已有相关法医学研究应用的Roche、Illumina和Life Technologies三大测序技术公司推出的二代测序技术平台为例,探讨其在法医遗传学中的应用现状及潜能。目前可依赖这些平台完成DNA水平遗传标记(SNP、STR)、RNA水平遗传标记(m RNA、micro RNA)及线粒体DNA全基因组的测序。然而,技术产品的推出及验证、分析软件的成熟化、与现有数据库的对接、大数据使用中的伦理问题等都是决定该技术能否替代(或补充)成熟PCR毛细管电泳技术以及普遍应用于案件检测的关键。     

High-throughput DNA sequencing of environmentally insulted latent fingerprints after visualization with nanoscale columnar-thin-film technique

The goals of this study were to (a) ascertain human identity capabilities of DNA obtained from latent fingerprints that have been first environmentally insulted and then developed by the deposition of a columnar thin film (CTF), and (b) to determine if the CTF process and material are detrimental to single nucleotide polymorphism (SNP) analysis. Fingerprints were deposited on five different types of substrates and aged for one day, 7 days or 30 days while being environmentally insulted under one of the four conditions: 16.6 °C and 60% relative humidity (RH) (Condition A), 24.5 °C and 60% RH (Condition B), 35 °C and 67% RH (Condition C) and a cold condition (Condition D). Then CTF technique was then on 59% of these fingerprints. DNA samples from 805 fingerprints were extracted, quantified, subjected to manual library preparation using the Precision ID Identity Panel, and underwent high-throughput sequencing. The Ion S5™ platform was employed to sequence 124 SNP amplicons. SNPs were successfully sequenced from 802/805 samples. Total read depth was consistent across environmental conditions, and majority of samples had 100% profile completeness and 100% concordance. Anecdotally, libraries that were amplified with a higher cycle number had more ‘Major Allele Frequency’ flags compared to samples amplified with 23 cycle numbers, possibly due to stochastic effects. Neither the substrates nor the CTF process and materials inhibit downstream DNA analysis. DNA of low quality and quantity from the chosen samples can be sequenced using the Precision ID Identity Panel on the Ion S5™ platform which performed well, however, a different approach may be needed if spurious alleles are suspected.     

The effectiveness of various strategies to improve DNA analysis of formaldehyde-damaged tissues from embalmed cadavers for human identification purposes

Formalin-fixed tissues provide the medical and forensic communities with alternative and often last resort sources of DNA for identification or diagnostic purposes. The DNA in these samples can be highly degraded and chemically damaged, making downstream genotyping using short tandem repeats (STRs) challenging. Therefore, the use of alternative genetic markers, methods that pre-amplify the low amount of good quality DNA present, or methods that repair the damaged DNA template may provide more probative genetic information. This study investigated whether whole genome amplification (WGA) and DNA repair could improve STR typing of formaldehyde-damaged (FD) tissues from embalmed cadavers. Additionally, comparative genotyping success using bi-allelic markers, including INDELs and SNPs, was explored. Calculated random match probabilities (RMPs) using traditional STRs, INDEL markers, and two next generation sequencing (NGS) panels were compared across all samples. Overall, results showed that neither WGA nor DNA repair substantially improved STR success rates from formalin-fixed tissue samples. However, when DNA from FD samples was genotyped using INDEL and SNP-based panels, the RMP of each sample was markedly lower than the RMPs calculated from partial STR profiles. Therefore, the results of this study suggest that rather than attempting to improve the quantity and quality of severely damaged and degraded DNA prior to STR typing, a more productive approach may be to target smaller amplicons to provide more discriminatory DNA identifications. Furthermore, an NGS panel with less loci may yield better results when examining FD samples, due to more optimized chemistries that result in greater allelic balance and amplicon coverage.     

平行进口的法律性质分析

平行进口是否构成侵权已成为知识产权国际保护中的热点问题。文章研究了平行进口的理论基础 ,分析了TRIPS协议对平行进口的态度和西方主要国家在此问题上的立法与实践 ,探讨了我国现行法律制度和经济发展现状 ,并在此基础上提出我国对平行进口不应采取过于严厉的态度。     

DNA甲基化检测方法及其法医学应用研究进展

DNA甲基化是表观遗传标记的重要组成部分,参与基因表达的调控,在生物发育、衰老以及肿瘤学等领域受到广泛关注。由于具有相对稳定性、可遗传、含量丰富、随龄变化等特点,在法医学领域,DNA甲基化可以作为DNA序列相关经典遗传标记的有效补充,用于年龄推断、组织体液来源检测、同卵双生子的鉴定等。DNA甲基化的检测方法主要有基于甲基化敏感限制性内切酶、重亚硫酸盐转化以及甲基化CpG结合蛋白等原理而建立的一系列方法,近年研究表明,第三代测序技术也可用于DNA甲基化检测。本文就DNA甲基化检测方法及在法医学领域的应用研究进行回顾与综述,以期为法医学实践提供参考。     

国际平行诉讼的成因与对策分析

分析了国际平行诉讼产生的原因 ,阐述并分析了各国国内及相关国际条约对国际平行诉讼采用的不同对策 ,就当前我国立法和司法现状指出了存在的问题并提出了建议。     

浅析知识产权平行进口问题

由于经济利益的驱动,在国际贸易中,一国未被授权的进口商从外国知识产权人或持有人手中购得商品并未经批准而输入本国,而该知识产权在此以前已在本国得到保护,从而产生平行进口的问题.知识产权平行进口是国际贸易领域不可回避的课题,其合法性问题始终是知识产权法律界争议的焦点之一.学者因为对权利穷竭与地域性原则的理解不同,因而理论界出现了两种截然相反的观点.通过对相关理论及实践的分析,提出平行进口的合法化取决于各国国内法对于权利穷竭原则与地域性原则的明确与具体化的观点,并针对中国入市后对外贸易发展的需要提出了相应对策.     

国际贸易中的平行进口与知识产权保护

平行进口作为目前国际上的一个热门话题,在专利、商标、著作权等知识产权领域广泛存在。由于世贸组织的TRIPS协议没有提出较为统一的原则,因此各国采取了不同立场。随着中国加入WTO,我国由平行进口引发的纠纷也日渐增多。我国在对待平行进口问题上,应区分专利权、商标权以及著作权,分别加以考虑。     

Aid and state-building,Part II: Afghanistan and Iraq

Part I of this article found that, in South Korea and Taiwan, institutional legacy and continuity as well as the politics of aid did matter for post-war state-building. The inheritance and continuity of Weberian states and the receipt of aid either as budget support or increasingly aligned with local priorities helped to foster state-building. Part II of the study in this article explores a different dynamic of post-war aid to Afghanistan and Iraq which had a legacy of neopatrimonial and weak states. It argues that under more adverse initial conditions – for a neopatrimonial state – the role of aid regime and state-building strategies become even more important. Under these conditions, aid and state-building strategies may undermine state-building if they induce discontinuity in the existing state capacity and create parallel institutions to those of the state. Depending on the policies, state weakness may be reinforced if leaders are preoccupied with the politics of patronage.