纳米磁珠提取法在骨骼DNA提取中的应用

目的对纳米磁珠法提取纯化骨骼DNA的效果进行比较评价,为方法选择提供应用参考。方法取泥土掩埋、水中浸泡1~10年不等的25根长骨,经水洗、刮净,液氮冷冻研磨器将骨骼研磨成粉末状,分别应用纳米磁珠提取法和King Fisher仪器自动化提取法提取DNA,IdentifilerPlus试剂盒进行扩增,ABI 3100遗传分析仪进行STR分型检测;对两种方法提取的DNA定量和经扩增、分型检测的结果进行比较。结果骨骼样本采用纳米磁珠法提取到的样本DNA(1.237 5ng/μL±0.319 2ng/μL),较之King Fisher法的浓度(0.506 2ng/μL±0.280 5ng/μL)更高,两种方法间差异具有统计学意义(P0.05);而纳米磁珠法的分型成功率亦更高,两种方法间差异具有统计学意义(P0.05)。结论用纳米磁珠提取纯化骨骼DNA,能得到高质量DNA模板,有利于提高分型检验的成功率,可在实际检案中选择使用。     

陈旧性骨骼DNA提取方法的探索

目的探索陈旧性骨骼DNA的提取方法。方法收集4~15年陈旧骨骼样本,去除表面污染物,经脱钙、裂解提取各样本DNA,使用QIAquickPCR Purification试剂盒进行纯化,检测DNA纯度和浓度,应用Power PlexFusion荧光标记复合扩增系统进行扩增,AB-3500型遗传分析仪检测STR分型。结果 15例陈旧性骨骼经提取、纯化,提取的DNA模板浓度较高,在42.9~176.4ng/μL之间,A260/A280值较稳定,在1.06~1.40之间;所有样本均获得完整STR分型。结论该方法简便快速,提取效果好,能够适用于法医学实际检案。     

2种陈旧骨骼、牙齿DNA纯化方法的比较

目的比较有机法+QIAquick纯化法和DNA IQ磁珠法对陈旧骨骼和牙齿DNA的纯化效果。方法选择10份陈旧骨骼和12份牙齿样本,进行消化后分别采用有机法+QIAquick纯化法和DNA IQ磁珠法进行提取纯化,进行DNA定量后用SinofilerTM试剂盒进行检测。结果 2种方法纯化的骨骼、牙齿DNA的IPC CT值无显著差异。有机法+QIAquick纯化法纯化的骨骼、牙齿DNA平均浓度分别为0.180ng/μL±0.068ng/μL和0.132ng/μL±0.027ng/μL,所有样品均获得全部基因分型。DNA IQ磁珠法纯化的DNA平均浓度分别为0.038ng/μL±0.028ng/μL和0.036ng/μL±0.007ng/μL,有5份骨骼和6份牙齿样本仅获得部分基因分型或未能分型。结论有机法+QIAquick纯化法对陈旧骨骼、牙齿DNA的纯化效果优于DNA IQ磁珠法。     

MiSeq FGx~(TM)系统进行基因突变亲权鉴定1例

<正>1材料与方法1.1 DNA提取及定量样本来源于本实验室日常案件中经ID-PLUS检测存在STR遗传位点突变的三联体的FTA唾液口腔卡,并使用Qiagen DNA Investigator试剂盒(Qiagen公司)提取样本DNA。样本DNA采用Qubit?3.0荧光定量仪(Life Technologies公司)进行DNA浓度检测,并将浓度稀释到0.2ng/μL[1]。     

05式警用转轮手枪弹壳接触DNA检验方法的比较

目的比较05式警用转轮手枪弹壳表面接触性DNA检验方法,为实际检验提供参考和借鉴。方法制备40例击发后手枪弹壳的模拟样本,分别用两步转移法提取弹壳表面不同部位检材,采用两种DNA提取法和两种扩增试剂盒对样本进行STR分型检验,比较评价检验结果。结果避开发射药残留区域采用两步转移法提取样本,有助于提高检出率;Chelex-100联合Microcon-100法提取模板DNA的产量最高可达1.18ng,高于Mini M48试剂盒法(0.91ng);MiniFilerTM试剂盒的等位基因检出率(23.61%)高于IdentifilerTM试剂盒(6.41%)。结论采用选择适当区域提取检材,采用Chelex-100联合Microcon-100法提取DNA,经MiniFilerTM试剂盒扩增,进行弹壳接触DNA分型的效果较好。     

荧光定量PCR技术验证Sinofiler试剂盒的适宜模板量

目的探讨Sinofiler试剂盒适合扩增的模板DNA用量。方法选取经Sinofiler试剂盒基因分型且图谱完整、扩增均衡性好的DNA样本,进行荧光定量PCR检测。结果实验结果表明,在12.5μL体系中,1.29～1.51ng的模板DNA能够获得理想的分型结果。结论应用荧光定量PCR技术检测不同试剂盒适合扩增的模板DNA用量是一种准确、有效的方法。     

PuriTyper~(TM)法医学纯化试剂盒的性能验证

目的验证PuriTyperTM纯化试剂盒各项性能指标和法医学应用价值。方法收集及制备抗凝血液、常见案件检材(唾液、烟头、精液、毛发、指甲、骨骼及组织块)、斑痕样本(血斑、唾液斑、精斑)以及模拟添加抑制剂和模仿自然环境中放置的血斑。采用PuriTyperTM纯化试剂盒提取纯化并进行DNA定量,IdentifilerTM复合扩增试剂盒扩增,产物经ABI 3130遗传分析仪进行检测,Genemapper软件分析结果,对该试剂盒灵敏度、稳定性、重复性、检材适应性进行测试。结果采用该试剂盒提取0.1～40μL血液分别获得0.042～26.45ng/μL的DNA。3种斑痕样本DNA产量平行试验结果稳定。不同类型检材重复检验所获IPC的CT平均值在27.60至28.03之间。常见案件检材所得分型与已知结果均一致。结论 PuriTyperTM纯化试剂盒能够满足法医DNA检验的要求,对法医学实践具有重要的应用价值。     

泥土中二氧化硅对硅珠法提取现场生物物证DNA的影响

目的 研究泥土中二氧化硅对硅珠法提取现场生物物证DNA的影响。方法 泥浆悬液和稀释血混合,制成混有灰尘、泥土的生物样本以模拟现场生物物证,分别用加热裂解和胍盐化学裂解的方式进行细胞裂解,硅珠法提取DNA后用Identifiler Plus试剂盒进行PCR扩增及毛细管电泳检测,并对电泳结果进行比较;用泥浆悬液代替硅珠提取稀释血DNA,反向验证泥土中二氧化硅对硅珠法提取现场生物物证DNA的影响。结果 混有泥浆悬液的4μL、20μL稀释血加热裂解,提取扩增后得到完整STR分型,平均峰高1 969.7±376.9 RFU、9 706.7±349.8 RFU;混有泥浆悬液的4μL稀释血胍盐化学裂解,提取扩增后无法获得完整STR分型;混有泥浆悬液的20μL稀释血胍盐化学裂解,提取扩增后得到完整STR分型,平均峰高1 899.8±801.3 RFU;泥浆悬液代替硅珠提取20μL稀释血得到完整STR分型。结论 泥土中的二氧化硅在胍盐存在条件下会与DNA结合,导致硅珠法提取现场生物物证DNA的回收效率降低。     

快速PCR仪与普通PCR仪扩增效果的比较研究

目的比较快速PCR仪与普通PCR仪的扩增效果。方法浓度为0.1、0.05、0.025、0.0125、0.0063ng/μL的9947A标准品各取样100例,采用Identifiler Plus试剂盒构建扩增体系,其中50例在普通PCR仪(9700型扩增仪)上进行扩增,50例在快速PCR仪(Speed cycler2扩增仪)上扩增,3500x L型遗传分析仪检测,对比每种浓度组两种扩增仪的检验结果。结果两种PCR仪的检出率均无差异(P0.05)。但当9947A的浓度为0.0125、0.0063ng/μL时,普通PCR仪检验样本的STR基因座检出数(13.7±1.0;11.3±1.5)均高于快速PCR仪(13.1±1.3;9.9±1.9),差异有统计学意义(P=0.029;P0.001);当9947A的浓度为0.1、0.05、0.025 ng/μL时,普通PCR仪检验样本的图谱峰高(18931±4625;13437±3165;5752±1344)均高于快速PCR仪(16929±4034;11815±4120;4865±1401),差异有统计学意义(P=0.023;P=0.030;P=0.002)。结论快速PCR仪可在较短的时间内达到与普通PCR仪相当的检出率,适合于实际案件中的应用;普通PCR仪检验获得的STR分型结果质量可能更高。     

生物检材DNA碱性裂解提取法的优化与改良

目的通过对DNA碱性裂解提取法进行优化和改进,建立一种操作更简便、检验快速、结果更可靠的生物检材DNA提取方法。方法以抗凝血作为检验样本,通过改变提取试剂的浓度、pH值、保存时间及孵化样本的温度、时间等条件,观测对检验结果峰值的影响,以确定提取试剂最佳条件,DNA提取物最佳保存条件。通过用改良后的碱性裂解法及Chelex100法同时提取不同量的抗凝血样本、各类法医生物检材,用不同厂家的DNA试剂盒扩增,对碱性裂解法的灵敏度、适应性和适用性进行评估。结果改良后的碱性裂解法只需将生物检材用NaOH（0.25mol/L）99℃孵化8min,振荡后加入TrisHCl（0.05mol/L,pH=6.5）中和液,离心后直接扩增检测。DNA提取物于-20℃冰箱可长期保存。对于毛发及精斑类检材优于Chelex100法。DNA提取物适用于现有各种DNA试剂盒扩增检测,其灵敏度与Chelex100法相似。结论改良后的碱性裂解法,操作简便、检验快速、结果可靠,可适合于法庭科学的检案与建库。     

Bhartṛhari and Maṇḍana on Avidyā

The concept of avidyā is one of the central categories in the Advaita of Śaṇkara and Maṇḍana. Shifting the focus from māyā, interpreted either as illusion or as the divine power, this concept brings ignorance to the forefront in describing duality and bondage. Although all Advaitins accept avidyā as a category, its scope and nature is interpreted in multiple ways. Key elements in Maṇḍana’s philosophy include the plurality of avidyā, individual selves as its substrate and the Brahman as its field (viṣaya), and the distinction in avidyā between non-apprehension and misapprehension. A closer investigation shows that Maṇḍana is directly influenced by Bhartṛhari’s linguistic non-dualism in developing the concept of avidyā. This study also compares other key constituents such as vivartta and pariṇāma that are relevant to the analysis of avidyā. As the concept of counter-image (pratibimba) emerges as a distinct stream of Advaita subsequent to Maṇḍana, this study also compares the application of pratibimba in the writings of Bhartṛhari and Maṇḍana.     

Unraveling the <Emphasis Type="Italic">Kāvyaprakāśa</Emphasis>: Jayadeva Pīyūṣavarṣa’s idiosyncratic sequence of topics in the <Emphasis Type="Italic">Candrāloka</Emphasis>

In his twelfth century alaṃkāraśāstra, the Candrāloka, Jayadeva Pīyūṣavarṣa reverses the sequence of topics found in Mammaṭa’s Kāvyapr-akāśa, an earlier and immensely popular work. With such a structural revisionism, Jayadeva asserts the autonomy of his own work and puts forth an ambitious critique of earlier approaches to literary analysis. Jayadeva investigates the technical and aesthetic components of poetry in the first part of the Candrāloka, prior to his formal semantic investigations in the latter half of the text, thus suggesting that aesthetic evaluations of poetry beneficially inform scientific investigations of language. Jayadeva’s organization of his chapters on the semantic operations, moreover, intimates that the study of suggestive and metaphoric functions of language clarifies our understanding of denotation, which is conventionally understood to be the primary and direct path of verbal designation.     

Immigration,intermarriage and the changing face of Europe in the post war period

In this article we compare the propensity to intermarry of various migrant groups and their children who settled in Germany, France, England, Belgium and the Netherlands in the post-war period, using a wide range of available statistical data. We try to explain different intermarriage patterns within the framework of Alba and Nee's assimilation theory and pay special attention to the role of religion, colour and colonial background. We therefore compare colonial with non colonial migrants and within these categories between groups with ‘European’ (Christian) and non-European (Islam, Hinduism) religions. First of all, religion appears to be an important variable. Migrants whose faith has no tradition in Western Europe intermarry at a much lower rate than those whose religious backgrounds correspond with those that are common in the country of settlement. The rate of ethnic endogamous marriages in Western Europe are highest in Hindu and Muslim communities, often regardless if they came as guest workers or colonial migrants. Whereas differences in religion diminish the propensity to intermarry, colour or ‘racial’ differences on the other hand seem to be less important. This is largely explained by the pre-migration socialisation. Furthermore, the paper argues that the attention to institutions, as rightly advocated by Richard Alba and Victor Nee, needs a more refined and layered elaboration. Institutions, often as barriers to intermarriage, do not only emanate from the receiving society, but also—be it less formalized—within migrant communities. Especially religions and family systems, but also organized nationalist feelings, can have a profound influence on how migrants think about endogamy. Finally, strong pressures to assimilate, often through institutionalized forms of discrimination and stigmatization, not only produce isolation and frustrate assimilation (with resulting low intermarriage rates), but can also stimulate assimilation by 'passing' mechanisms. These factors, together with a more comparative perspective, are not completely ignored in the new assimilation theory, but—as this study of Western European intermarriage patterns stresses—deserve to be included more systematically in historical and social scientist analyses.     

The Supreme Array Scripture: A New Interpretation of the Title “Gaṇḍavyūha-sūtra”

This article argues for a new interpretation of the Sanskrit compound gaṇḍa-vyūha as it is used in the common title of the Mahāyāna text the Gaṇḍavyūha-Sūtra.The author begins by providing a brief history of the sūtra’s appellations in Chinese and Tibetan sources. Next, the meanings of gaṇḍa (the problematic member of the compound) are explored. The author proposes that contemporary scholars have overlooked a meaning of gaṇḍa occurring in some compounds, wherein gaṇḍa can mean simply “great,” “big” or “massive.” This general sense is particularly common in the compound gaṇda-śaila (a “massive rock” or “boulder”) and is found in such texts as the Bhāgavata Purāṇa, the Harivaṃśa and the Harṣacarita. Following the discussion of Gaṇḍa, the author examines the term vyūha (“array”) as it is used in the Gaṇḍavyūha-sūtra. The article concludes with the suggestion that a more appropriate translation of the Gaṇḍavyūha-sūtra would be “The Supreme array Scripture.”     

Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA

Luminol, leuchomalachite green, phenolphthalein, Hemastix, Hemident, and Bluestar are all used as presumptive tests for blood. In this study, the tests were subjected to dilute blood (from 1:10,000 to 1:10,000,000), many common household substance, and chemicals. Samples were tested for DNA to determine whether the presumptive tests damaged or destroyed DNA. The DNA loci tested were D2S1338 and D19S433. Leuchomalachite green had a sensitivity of 1:10,000, while the remaining tests were able to detect blood to a dilution of 1:100,000. Substances tested include saliva, semen, potato, tomato, tomato sauce, tomato sauce with meat, red onion, red kidney bean, horseradish, 0.1 M ascorbic acid, 5% bleach, 10% cupric sulfate, 10% ferric sulfate, and 10% nickel chloride. Of all the substances tested, not one of the household items reacted with every test; however, the chemicals did. DNA was recovered and amplified from luminol, phenolphthalein, Hemastix, and Bluestar, but not from leuchomalachite green or Hemident.     

Illumination,Imagination, Creativity: Rājaśekhara,Kuntaka, and Jagannātha on <Emphasis Type="Italic">Pratibhā</Emphasis>

Sanskrit poeticians make the visionary faculty of pratibhā a necessary part of the professional poet’s make-up. The term has a pre-history in Bhartṛhari’s linguistic metaphysics, where it is used to explain the unitary perception of meaning. This essay examines the relation between pratibhā and possible theories of the imagination, with a focus on three unusual theoreticians—Rājaśekhara, Kuntaka, and Jagannātha Paṇḍita. Rājaśekhara offers an analysis of pratibhā that is heavily interactive, requiring the discerning presence of the bhāvaka listener or critic; he also positions pratibhā in relation to Bildung (vyutpatti) and practice. For Kuntaka, pratibhā, never an ex nihilo creation by a poet, serves as the basis for the peculiar forms of intensified insight and experience that constitute poetry; these may also involve the creative scrambling and re-articulation of the object in terms of its systemic composition. At times, Kuntaka’s pratibhā comes close to a strong notion of imaginative process. But the full-fledged thematization of the imagination, and of pratibhā as its support and mechanism, is best seen in the seventeenth-century debates preserved for us by Jagannātha. A link is suggested between the discourse of poetic imagination in Jagannātha and similar themes that turn up in Indo-Persian poets such as Bedil.     

Preventing Homelessness for System‐Involved Youth

Too many youth and young adults find themselves on the streets, couch‐surfing with friends, in emergency shelters or worse, after exiting the child welfare and juvenile justice systems. In some circumstances, youth have had court hearings until their exit from the legal system, but those hearings have not focused on long‐range plans of youth and emergencies youth may encounter. In other circumstances, there has been little or no planning prior to discharge, especially for young people who leave the juvenile justice system. Courts can and should prevent, alleviate or end youth homelessness for youth who appear before them through strategies that are enumerated in the recently‐passed NCJFCJ resolution. This article expounds on three of these strategies – coordinating transition and re‐entry plans, insisting on effective legal representation of youth, and utilizing sound judicial leadership. It also describes the concurrent efforts of the Coalition for Juvenile Justice and the American Bar Association's Homeless Youth Legal Network to remove legal barriers and improve outcomes for youth and young adults experiencing homelessness.     

Best practices for structuring a family court parenting plan evaluation under the 2022 AFCC Guidelines

Evaluation appointment orders provide enforceable scaffolding for conduct of family court parenting plan evaluations, and use of the evaluator's reports, feedback, file, and testimony. Unlike a contract, a stipulated or adjudicated appointment order is directly enforceable by the family court. It unambiguously positions the evaluator as the family court's appointee – answerable directly to the court and, in some jurisdictions, protected by quasi-judicial immunity from damages claims. A well-crafted appointment order governs the roles and expectations of the court, the evaluator, the parties, the lawyers, and the collateral witnesses. An appointment order mandates the legal duties, rights, powers, and responsibilities of the professionals, the parties, and the collateral witnesses. At minimum, an appointment order articulates the legal basis for the appointment, the purpose and scope of the evaluation, compensation of evaluator, and the duty of the parties to participate in the process. A written evaluation protocol or procedures statement discloses in advance the methods of investigation and assessment that the evaluator intends to use. Together, the appointment order and written protocol help the evaluator, lawyers, parents, and judge manage the complexity of the evaluation process.     

Monks Who Have Sex: Pārājika Penance in Indian Buddhist Monasticisms

In the study of Buddhism it is commonly accepted that a monk or nun who commits a pārājika offence is permanently and irrevocably expelled from the Buddhist monastic order. This view is based primarily on readings of the Pāli Vinaya. With the exception of the Pāli Vinaya, however, all other extant Buddhist monastic law codes (Dharmaguptaka, Mahāsāṅghika, Mahīśāsaka, Sarvāstivāda and Mūlasarvāstivāda) contain detailed provisions for monks and nuns who commit pārājikas but nevertheless wish to remain within the saṅgha. These monastics are not expelled. Rather, they are granted a special status known as the śikṣādattaka. In this paper I explore the rules. concerning pārājika penance and the śikṣādattaka with specific regard to monastic celibacy. Given that five out of six extant law codes recognise this remarkable accommodation to the rule of celibacy, I argue that we must look to Vinayas other than the Pāli Vinaya if we are to arrive at a nuanced and representative view of Indian Buddhist monasticism.

Grant me chastity and continence, but not yet. Augustine of Hippo (354–430 C.E.)

     

What similes in Sāṃkhya do: a comparison of the similes in the Sāṃkhya texts in the Mahābhārata, the Sāṃkhyakārikā and the Sāṃkhyasūtra

In Sāṃkhya similes are an important means to communicate basic philosophical teachings. In the texts similes are frequently used, especially in the Sāṃkhya passages in the Mahābhārata, in the Sāṃkhyakārikā and in the Sāṃkhyasūtra. This paper compares the similes in these three texts and analyses changes in the philosophy as revealed in the similes. A comparison of the similes of Sāṃkhya texts produced over more than one thousand years reveals changes in the emphasis in this philosophical system. The purpose of the similes in the Sāṃkhya passages of the Mahābhārata is to produce an intuitive understanding of the separateness of puruṣa and prakṛti. The similes are designed to lead the listener to understand this basic dualism. In the Sāṃkhyakārikā the most difficult issues are the relationship between prakṛti and puruṣa and the idea of prakṛti working for the salvation of puruṣa. One whole chapter of the Sāṃkhyasūtra is devoted to similes.