mRNA在体液斑鉴定研究中内参基因的选择与应用

利用mRNA分析技术鉴定生物检材中各种体液斑迹的组织属性来源是近年来法医物证学的一个重要进展。终点逆转录PCR和实时荧光定量PCR技术是法医学实验室常用的mRNA检测技术,为保证实验结果的准确性,研究者通常选用组成性表达的管家基因作为内参,对不同样本提取RNA进行质控和标准化,以正确评估目标mRNA在各样本中的表达量。本文聚焦近年来使用mRNA分析技术进行体液斑迹组织属性来源鉴定的研究报道,对这些工作中内参基因的选择与应用效果进行综述。     

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。     

血痕特异性mRNA标记研究

血痕是案发现场尤其是命案中比较常见的生物物证,而血痕的正确组织来源推断是当前鉴定工作中急需解决的问题。随着法医物证学的不断发展,以mRNA(messenger RNA)为基础的体液斑迹组织来源鉴定技术作为一种不同于传统血痕免疫学检测的新型方法,已经越加显示其独特的优越性。在该技术的基础上,实现共同提取生物物证的RNA与DNA的目标,建立体液斑迹鉴定与DNA分型兼容的方法,有利于现场重建,提高生物物证的证据效力,完善证据链。本研究建立了一个包括血痕总RNA提取、逆转录、荧光特异引物扩增、遗传分析仪电泳检测分析等步骤的血痕来源推断技术平台。实验共采集制备了40份的中国人群(女性)外周血、16份月经血样本,筛选了5个外周血标记:HBA、HBB、GYPA、SPTB、ALAS2,2个月经血标记:MMP7、MMP11,构建了一个囊括外周血、月经血特异标记的荧光复合扩增体系。结果显示mRNA技术为基础的鉴定血痕来源的方法是可行的,并且建立了血痕RNA检验的遗传分析仪结果判读方法。     

基于微生物标记物鉴别唾液、阴道分泌物的多重荧光标记复合扩增检测体系构建

目的 构建基于微生物标记物的荧光复合检测体系，实现对唾液斑迹、阴道分泌物斑迹的区分鉴别。方法经文献检索结合本实验室前期工作，根据特异性和丰度筛选唾液和阴道分泌物微生物标记物；制备唾液斑迹、阴道分泌物斑迹样本，提取总DNA，利用毛细管电泳技术构建基于微生物标记物鉴别唾液、阴道分泌物的多重荧光标记复合扩增检测体系，并使用体液斑迹样本对体系特异性、灵敏度进行检验。结果 本研究筛选出5个唾液微生物标记物（微黄奈瑟氏菌Neisseria subflava、非典型韦荣氏菌Veillonella atypica、唾液链球菌Streptococcus salivarius、口腔链球菌Streptococcus oralis、凯托卟啉单胞菌Porphyromonas catoniae）和3个阴道分泌物微生物标记物（卷曲乳杆菌Lactobacillus crispatus、加氏乳杆菌Lactobacillus gasseri、短双歧杆菌Bifidobacterium breve），利用毛细管电泳技术构建了8重微生物荧光标记复合扩增检测体系，并使用140份样本对体系特异性进行验证；测试结果显示，唾液和阴道分...     

基于microRNA表达谱初步构建PLS-DA体液识别模型

针对外周血、唾液、精液、月经血、阴道分泌物这五种法医学常见的体液样本，采用二代测序（NGS）平台检测获得microRNA(miRNA)表达谱，使用偏最小二乘判别分析（PLS-DA）建立基于这五种体液的识别模型，并探讨PLS-DA在法医体液溯源中的应用价值。经优化小分子RNA文库制备过程，采用Ion Torrent S5 XL测序系统对前述法医学五种体液样本（每种10例）进行小RNA测序，以PLS-DA构建体液识别模型，评估不同数量miRNA标记组合下预测的准确性。本研究获得法医学常见五种体液样本的miRNA表达谱，外周血与月经血中表达量前10名的miRNAs有6个重叠；唾液和阴道分泌物中表达量前10名的miRNAs有4个重叠。基于全数据集、107个和11个miRNAs构建的体液来源识别模型的准确率分别为0.95、0.94、0.89。本研究通过NGS测序分析获得了五种体液样本的miRNA组（miRNome），利用PLS-DA初步构建了体液识别模型，对于应用miRNome进行体液识别的相关研究具有参考价值。     

miRNA451用于法医学血液鉴别的可行性研究

目的探讨miRNA451是否可以用于法医学血液类物证的鉴别。方法以法医学常见血液类(外周血和月经血)及非血液类(精液、唾液、阴道分泌液)体液样本为研究对象,提取总RNA,以RT-qPCR方法检测miRNA451的表达,利用SPSS 22.0进行统计分析。结果 miRNA451在外周血及月经血中均具有高表达的特点,且表达量显著高于在唾液、精液及阴道分泌液等非血液类物证中的表达量(6组P均0.05);放置一年的血斑中miRNA451表达量与新鲜血液无显著差异。结论 miRNA451在血液类体液样本中的表达水平显著高于非血液类体液样本,且稳定性良好,可作为法医学血液类体液样本鉴别的良好分子标记物。     

利用mRNA检验确证无精子症精斑1例

正近年来,利用mRNA进行生物物证体液斑迹的研究已经取得一定的进展,其在DNA鉴定工作中发挥的作用也越来越被重视。本实验室受理了1起强奸案,通过mRNA的检测分析,为刻画嫌疑人提供了依据,现报道如下。1案例资料1.1简要案情2015年4月某强奸案中,现场提取到有疑似精斑的床单1件,斑迹经人精P30试纸条检测为阳性。用差异裂解法提取检材DNA,同时检测上清液和沉     

法医体液溯源的分子生物学技术研究进展

通过对犯罪现场发现的体液溯源,可以在样本来源者与实际犯罪行为之间建立联系,为现场重建提供重要证据。随着分子生物学技术的发展,DNA甲基化、RNA等遗传标记以及特定微生物检测被发现具有潜在的法医体液溯源价值。本文对近年来应用分子生物学方法进行法医体液溯源的研究现状和应用前景进行综述。     

ARMS-PCR技术检测法医学体液样本DNA甲基化方案

目的 案件现场遗留的体液等生物检材对案件性质的判定以及犯罪现场的重建具有重要意义。本研究旨在确定ARMS-PCR技术能否用于DNA甲基化位点的检测，评估候选的DNA甲基化标记物是否具有体液特异性，以便于解决目前体液鉴定复杂、成本高、混合样本识别困难等问题。方法 利用ARMS技术设计DNA甲基化特异性引物，构建复合扩增体系，通过毛细管电泳进行分型检测。结果 本研究利用ARMS技术以及毛细管电泳构建了包含22个甲基化位点的复合扩增体系，进一步检测并识别出了100例单一体液样本。结论 该方法可实现甲基化位点的检测，是一种快速、低成本的检测方式，候选位点具有体液特异性。     

应用SNaPshot技术检测精液特异性cSNP遗传标记

目的 探讨基于SNaPshot技术的精液特异性编码区单核苷酸多态性（codingregionsinglenucleotidepolymorphism,cSNP）遗传标记检测在精液（斑）溯源及混合体液（斑）鉴定中的可行性。方法 制备16例精液斑和11例精液-静脉血混合斑样本,提取其基因组DNA(genomicDNA,gDNA)和总RNA,并将总RNA逆转录为互补DNA(complementaryDNA,cDNA)。在已验证的精液特异性mRNA编码基因上筛选cSNP遗传标记。基于SNaPshot技术构建cSNP复合检测体系并通过CE对样本进行基因分型。结果 成功构建了包含5个精液特异性cSNP的复合检测体系。在16例精液样本中,除了位于TGM4基因上的cSNP在cDNA的检测结果中出现等位基因丢失外,其余cSNP的gDNA和cDNA分型结果高度一致。检测精液-静脉血混合斑时,检出的cSNP分型结果均与精液提供者的基因型一致,未受到静脉血提供者基因型的干扰。结论应用SNaPshot技术检测精液特异性cSNP的方法可以应用于法医学精液（斑）的基因分型,并为混合体液（斑）中精液来源个体的判定提供...     

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.     

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.     

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.     

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.     

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.     

<Emphasis Type="Italic">Anyathākhyāti</Emphasis>: A Critique by Appaya Dīkṣita in the <Emphasis Type="Italic">Parimala</Emphasis>

In this paper, the problem of illusory perception, as approached by the Nyāya and Advaita Vedānta schools of philosophy, is discussed from the standpoint of the Parimala. This seminal work belonging to the Bhāmatī tradition of Advaita Vedānta was composed in the sixteenth century by the polymath Appaya Dīk?ita. In the context of discussing various theories of illusion, Dīk?ita dwells upon the Nyāya theory of anyathākhyāti, and its connection with jñānalak?a?apratyāsatti as a causal factor for perception, and closely examines if such an extraordinary (alaukika) perception is tenable to explain illusory perception. He then proceeds to point out the deficiencies of this model and thereby brings to the fore the anirvacanīyakhyāti of Advaitins as the only theory which stands scrutiny.     

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.)