一种基于FRET的PSA荧光探针的制备及法医学应用

目的设计合成靶向识别检测前列腺特异性抗原的有机小分子荧光探针,探索荧光探针在生物物证精液斑确证试验上的应用。方法基于荧光共振能量转移机理设计合成前列腺特异性抗原荧光探针,通过质谱、高效液相色谱法等对探针进行结构及纯度表征分析,采用荧光光谱法测试探针对前列腺特异性抗原的荧光响应,制作不同附着物的精液样品检材,利用倒置荧光显微镜观察记录荧光成像。结果成功设计合成了一种前列腺特异性抗原荧光探针,荧光光谱实验表明其对前列腺特异性抗原有较好响应,最低检测限为0.8μg/mL,不同附着物精液检材荧光成像实验进一步验证了荧光探针在精液斑确证试验上的应用。结论本研究设计合成了一种对前列腺特异性抗原响应灵敏的荧光探针,有望用于法医学领域精液斑的确证试验。     

等位基因特异性PCR技术及其法医学应用

等位基因特异性PCR（allele-specific polymerase chain reaction,AS-PCR）是一种基于等位基因特异性引物引导的PCR技术,可以有效地分析单核苷酸多态性（SNP）,包括碱基的转换、颠换以及插入/缺失多态性,在疾病研究、分子诊断以及法医物证学研究中具有很好的应用价值.本文系统地综述了AS-PCR技术的原理、检测手段、改进方法及在常染色体、Y染色体和线粒体SNP等领域的研究成果,探讨其法医学应用价值.     

改良法检测微量血痕Gm抗原

Gm抗原为Grubb于1956年首次报道,其后相继发现了一系列密切相关的Gm抗原,讫今已叙述过的这类抗原至少有24种之多。世界卫生组织在1965年对Gm命名进行了修正,即是用数字来表示这一系统的有关抗原。由于Gm抗原是共显性的常染色体连锁等位基因的遗传产物,因而其表现型呈复染的遗传多态性。Gm抗原存在于     

性侵犯案件中对于精斑试纸条使用灵敏度以及特异性的研究

抗原胶体金免疫试纸条也就是精斑试纸条,这是一种使用色谱层析技术以及免疫学方法通过前列腺特异性抗原对特异性进行检测的办法.该办法在实际操作过程中具备简单、所耗费时间短、得出的结果容易判断以及没有特殊设备要求等诸多优势而在日常精斑确证试验中在法医学的实际检案上.在实际检测过程中,很可能会出现精子检测不到或者是男性的DNA不能进行扩增情况的发生,为对此种办法的灵敏度和特异性进行验证和比较,本文特对此课题做一探讨,相信本文能够对相关课题研究起到抛砖引玉的作用.     

SMCY性别特异融合抗原的克隆表达及其抗体制备

目的利用蛋白检测的快速性优势,研究不同性别在SMCY抗原氨基酸序列的差异,筛选出特异性氨基酸序列,并克隆表达性别特异融合抗原,制备相应抗体,建立一种快速鉴别法医物证性别的方法。方法通过对人SMCY和SMCX进行序列分析,发现了三段差异片段,采用搭桥PCR方法获得差异片段全长基因,连接入p ET-28a载体进行原核表达,用Ni柱纯化后的性别特异融合抗原免疫制备多克隆抗体,用ELISA法和western blot检测SMCY多抗与抗原的反应特异性,制作胶体金试纸条检测样本。结果筛选出具有性别特异性的氨基酸序列,获得SMCY性别特异融合抗原,成功制备出多克隆抗体及胶体金试纸条。结论获得SMCY性别特异融合抗原具有很好的抗原活性,制备的多克隆抗体可以与抗原特异性地结合,用于性别鉴定。     

DNA甲基化标记检测方法及其法医学应用

DNA甲基化是重要的表观遗传修饰,主要在转录水平上调控基因的表达。DNA甲基化在人类基因组中含量丰富、分布广泛;甲基化谱有时空特异性、细胞特异性和亲源特异性。新近研究表明DNA甲基化在组织体液鉴定、同卵双生子鉴定、年龄推断、性别推断和亲子鉴定等方面具有一定的应用价值,有望成为一种新的法医学遗传标记。本文就DNA甲基化检测方法的研究进展及法医学应用进行综述,以期为法医学实践提供参考。     

HLA DRB基因SSO分型方法的研究

按照第11届国际组织相容性抗原研讨会（IHW）工作会议HLAⅡ类PCR－SSO分型标准和美国国立骨髓供者计划组织（NMDP）对HLADRB基因分型要求,设计合成一对引物,可同时扩增HLA-DRB1＋B3＋B4＋BSDNA片段,长度为256bp,设计合成不同片段大小探针27种,可检出DRB座位上DRB1的39种等位基因,DRB3的3种等位基因,DRB4的1种等位基因和DRB5的3种等位基因。     

抗人SMCY抗原胶体金试纸条的研制

目的建立一种快速检测SMCY抗原的方法。方法采用胶体金免疫层析技术双抗体夹心检测法,在金标垫上包被胶体金标记的SMCY兔多抗,制成胶体金试纸条,用于检测12份猪、牛、狗、鸡和小鼠等动物血清以及50份人血清。结果制作的胶体金试纸条检测人和常见动物血清具有明显的种属特异性,能够区分男女血清。结论该方法可尝试用于鉴别男女血清,并具有一定种属特异性,其为法医学快速鉴别人类样本性别提供了研究方向。     

HLA基因多态性及其法医学应用

人类白细胞抗原(Human Leukocyte Antigen,HLA)基因复合物定位于人类第6号染色体短臂上,是人类最复杂的显性多态遗传系统,基因型可达108种之多,是理想的人类遗传标记,已广泛应用于法医学亲子鉴定和个体识别。本文就其基因多态性的研究进展和在法医学中的应用作一综述。     

闽南地区汉族17个Y-STR基因座遗传多态性

正在法医遗传学研究领域,Y染色体遗传标记已成为常染色体的重要辅助遗传标记。由于Y染色体遗传标记的遗传特点和在不同群体中的差异,其单倍型的分布具有群体特异性。本研究对中国闽南地区汉族17个Y-STR基因座和单倍型频率的遗传多态性进行调查,旨在为人类遗传学、法医遗传学研究提供群体遗传学数据。     

A test of social learning and intergenerational transmission among batterers

This research examined the direct and indirect transmission of family-of-origin violence among a sample of male domestic violence offenders. Intergenerational transmission of violence was tested by examining the effects of childhood corporal punishment experiences and witnessing inter-parental physical violence on the odds of reporting minor and severe intimate partner violence perpetration in adulthood. Social learning mechanisms were applied to examine the relationship between abuse experiences and the incidence of minor and severe forms of intimate partner violence. Use of a sample of 204 male domestic batterers attending court-mandated family violence intervention programs in an urban setting revealed considerable variation in minor and severe intimate partner violence. Results from logistic regression models suggested intergenerational transmission and social learning provided distinct mechanisms for both minor and severe forms of intimate partner violence.     

The Gender Gap in Sex Offender Punishment

This paper tests theoretical arguments that suggest court actors hold gendered views of sex offenders that result in a gender gap in sex offender punishment, where women who commit sexual offenses are treated more leniently than their male counterparts. We test this argument with precision matching analyses using 15 years of data on all felony sex offenders sentenced in a single state. Results indicate that gender disparities in sex offender sentencing exist and are pervasive across sex offense types. Specifically, male sex offenders are more likely to be sentenced to prison, and given longer terms, than female sex offenders. Findings are similar across sex offense severity and whether the offense involved a minor victim. These findings suggest that female sex offenders are treated more leniently than their matched male counterparts, even in instances of more serious sex offenses and those involving minor victims. Findings support theoretical arguments that contend that court decision-making is influenced by legally-irrelevant characteristics and raise questions about the source of gendered views of sex offenders and their effects on punishment approaches. Findings also raise questions about the virtue of get-tough sentencing policies that provide leeway for such dramatic variation across different groups of people.     

Static screening instruments for risk of minor and major violence

Background: Screens for violent convictions that are simple, accessible and parsimonious are needed, as a first stage in identifying those at high risk for further assessment. Aims: To construct and validate screening tools for minor and major violence convictions for released prisoners. Methods: Internal validation sample of 1647 serious offenders and an external validation of 46,704 general prisoners. The outcomes were binary indicators for having at least one conviction for minor and major violence. Risk factors were convictions for violence and age. Results: In the external validation sample, the instrument for risk of minor violence (PMIV) identified correctly 60.8% of male and 66.2% of female general prisoners. For risk of major violence, the instrument (PMAV) identified correctly 68.0% of male and 79.3% of female prisoners. Conclusions: The PMIV and PMAV will efficiently assist practitioners in a first stage of screening before in-depth clinical assessment of risk for future violent convictions.     

Prison Violence,Gender, and Perceptions: Testing a Missing Link in Discretion Research

Criminal justice literature often suggests female social control agents (e.g., police, prison staff, and others) use force less often than male colleagues facing similar situations. Most explain this with reference to personal or social factors which are gendered, arguing role-pressures or differences in skill sets between men and women lead to different reactions by agents. However, part of the story may also be about assessment rather than reaction—men and women may diverge in how they recognize and interpret cues in some (particularly minor) violence situations. Testing this, survey data was drawn from a random sample of 2,077 staff working in all 112 Federal Prisons operating in 2005. By estimating fixed effects models (grouped on prison), as well as controlling for correlates of exposure within a given prison, male and female staff were compared with respect to their assessment of statistically similar violence events. The data showed that women assessed fewer minor assault events to have occurred than their male colleagues, yet the same number of serious violence events. This supports the argument that perceptions may play an important role in explaining gender differences in the use of discretion.     

Recommendations for Correctional Leaders to Reduce Boundary Violations: Female Correctional Employees and Male Inmates

The correctional profession continues to report boundary-violating behavior by correctional employees with inmates, such as aiding an inmate in an escape and engaging in sexual contact with an inmate. These criminal behaviors obviously threaten the safety within the institutions and the community; however, these types of actions are normally preceded by minor boundary crossings within the institution. Therefore, all types of boundary violations and crossings between an inmate and an employee must be examined and eliminated. This article offers correctional leaders recommendations for organizational change that may reduce the number of boundary violations and crossings between female employees and male inmates. These recommendations are built on a larger qualitative study that used portraiture methodology, by interviewing 4 former female correctional employees who developed relationships with male inmates.     

Men’s Avoidance Coping and Female Partner’s Drinking Behavior: A High-Risk Context for Partner Violence?

The current study explored whether men's avoidance coping in response to the drinking behavior of their female partner with an alcohol use disorder (AUD) would be associated with higher levels of men's perpetration of intimate partner violence (IPV). Women with an AUD (n = 109) and their male partners in a U.S. urban area were assessed on men's perpetration of minor and severe violence using the Conflict Tactics Scale, men's avoidance coping using the Spouse Behavior Questionnaire, and men's and women's drinking behavior using the Time Line Follow Back Interview. Using multiple regression analysis, results showed that men's use of avoidance coping significantly predicted male IPV perpetration over and above the women's perpetration of violence toward him, while women's alcohol use did not significantly predict male-to-female IPV perpetration. Implications for teaching emotion-regulation strategies to male partners of women with an AUD to cope with partner drinking are discussed.     

Internal Validation of the AmpFlSTR Yfiler™ Amplification Kit for Use in Forensic Casework

Abstract:  Y-chromosomal short-tandem repeat (Y-STR) amplification has been used in forensic casework at the Bureau of Criminal Apprehension (BCA) Forensic Science Laboratory since 2003. At that time, two separate amplifications were required to type the SWGDAM recommended loci (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, and DYS439). The Yfiler™ kit coamplifies these loci as well as DYS437, DYS448, DYS456, DYS458, DYS635, and Y GATA H4. The Yfiler™ kit was validated following the internal validations outlined in the SWGDAM revised validation guidelines. Our studies show that 0.125 ng of male DNA will generate a complete 17 locus profile and that as little as 0.06 ng of male DNA yields an average of nine loci. In the male–male mixtures, a complete profile from the minor component was detected up to 1:5 ratio; most of the alleles of the minor component were detected at a 1:10 ratio and more than half the alleles of the minor component were detected at a 1:20 ratio. Complete YSTR profiles were obtained when 500 pg male DNA was mixed with female DNA at ratios up to 1:1000. At ratios of 1:5000 and 1:10,000 (male DNA to female DNA) inhibition of the YSTR amplification was evident. The YSTR results obtained for the adjudicated case samples gave significantly more probative information than the autosomal results. Our studies demonstrate that the Yfiler™ kit is extremely sensitive, does not exhibit cross-reactivity with female DNA, successfully types male DNA in the presence of overwhelming amounts of female DNA and is successful in typing actual forensic samples from adjudicated cases.     

Internal validation of the AmpFlSTR Yfiler amplification kit for use in forensic casework.

Y-chromosomal short-tandem repeat (Y-STR) amplification has been used in forensic casework at the Bureau of Criminal Apprehension (BCA) Forensic Science Laboratory since 2003. At that time, two separate amplifications were required to type the SWGDAM recommended loci (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, and DYS439). The Yfiler kit coamplifies these loci as well as DYS437, DYS448, DYS456, DYS458, DYS635, and Y GATA H4. The Yfiler kit was validated following the internal validations outlined in the SWGDAM revised validation guidelines. Our studies show that 0.125 ng of male DNA will generate a complete 17 locus profile and that as little as 0.06 ng of male DNA yields an average of nine loci. In the male-male mixtures, a complete profile from the minor component was detected up to 1:5 ratio; most of the alleles of the minor component were detected at a 1:10 ratio and more than half the alleles of the minor component were detected at a 1:20 ratio. Complete YSTR profiles were obtained when 500 pg male DNA was mixed with female DNA at ratios up to 1:1000. At ratios of 1:5000 and 1:10,000 (male DNA to female DNA) inhibition of the YSTR amplification was evident. The YSTR results obtained for the adjudicated case samples gave significantly more information than the autosomal results. Our studies demonstrate that the Yfiler kit is extremely sensitive, does not exhibit cross-reactivity with female DNA, successfully types male DNA in the presence of overwhelming amounts of female DNA and is successful in typing actual forensic samples from adjudicated cases.     

Performance characteristics of commercial Y-STR multiplex systems

In this work, a number of performance checks were carried out to evaluate the efficacy of commercial Y-short tandem repeats (Y-STR) kits for casework applications. The study evaluated the sensitivity, specificity and stability of the Y-STR markers used and the ability to obtain a male profile from postcoital samples taken at various time points after intercourse. All systems performed well with 1-3 ng of male DNA as recommended by the manufacturers. All systems gave full profiles at 100 pg of input DNA, which is within the realm of low copy number DNA analysis. Moreover all, except Y-Plex12, gave full profiles with 30-50 pg of male DNA. No increased performance was obtained with any of the systems by increasing the cycle number beyond that recommended by the various manufacturers. When up to 1 microg of female DNA was used (in the absence of male DNA) no female DNA cross reactivity was observed with the Y-Plex 12 and Y-Filer systems. PowerPlex Y produced female DNA derived products near the DYS438 and within the DYS392 loci at a rare allele position with high input DNA levels (300 ng and 1 microg, respectively). Male/female DNA admixture experiments indicated the particularly high specificity of the Y-Filer and PowerPlex Y systems under conditions of several thousand fold female DNA excess. All systems were able to detect the minor alleles in male/male DNA admixtures at a 1:5 dilution with the PowerPlex Y and Y-Filer being able to detect some minor alleles at 1:20. Species testing indicated some limited, minor cross reactivity of the commercial systems with some domestic male mammals although it is easily recognizable and would not pose any problems in casework analysis. As expected a significant number of cross-reacting products were obtained with nonhuman primate species. All Y-STR multiplex systems tested were able to produce complete Y-STR profiles from bloodstains and semen stains exposed up to 6 weeks when the samples were protected against precipitation and sunlight. However, exposure of the samples to precipitation either in the presence or absence of sunlight resulted in Y-STR profile loss over time, with total profile loss occurring with all systems after 3 weeks or more. Complete Y-STR profiles of the male donors up to 72 h postcoitus were obtained with all of the multiplex systems tested, except for Y-Plex12, which gave partial profiles.     

Death from renal cyst: spontaneous or traumatic rupture?

We report an unusual death of an apparently healthy 56-year-old male due to massive hemorrhage caused by rupture of an artery in the wall of a solitary renal cyst, possibly after a relatively minor trauma. Fatalities due to spontaneous or posttraumatic rupture of renal cysts are extremely rare but can represent a potential danger for people with acquired cystic kidney disease. Our report describes results of the forensic investigation and discusses possible mechanisms of the rupture.