Analysis of 50 SNPs in CYP2D6, CYP2C19, CYP2C9, CYP3A4 and CYP1A2 by MALDI-TOF mass spectrometry in Chinese Han population

One of the major challenges in the near future is the identification of genes that affect the metabolism of different drugs. Large scale association studies that utilise single nucleotide polymorphisms (SNPs) have been considered a valuable tool for this purpose. CYP2D6, CYP2C19, CYP2C9, CYP3A4 and CYP1A2 were found to be involved in the majority of hepatically cleared drugs. To determine the allele frequencies of some SNPs that may have great potential value in forensic science, we screened 50 SNPs in these 5 CYP genes in Chinese Han people using an accurate, high-throughput, cost-effective method. Primers were designed using the MassARRAY Assay Design software. Genomic DNA was prepared from blood samples obtained from individuals of Chinese Han origin. Multiplex PCR was performed to amplify the relevant gene fragments, and the polymorphisms were analysed by allele-specific primer extension followed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). A panel of genomic DNA samples previously genotyped by other methods were analysed simultaneously for quality control, and the results demonstrated that this assay was 100% accurate. A total of 17 of the analysed SNPs were polymorphic. Of these 17 SNPs, 8 (rs16947, rs28371725, rs1800754, rs4244285, rs4986893, rs12248560, rs3758580, rs2242480) had an allele frequency that was significantly different between this Chinese Han population and Caucasians (p<0.01). In addition, the frequencies of two of these SNPs (rs1800754, rs3758581) in our Chinese Han population differed significantly from the existing Chinese frequency data (p<0.01). The described method thus provides reliable results and enables the genotyping of up to thousands of samples by taking advantage of the high-throughput MALDI-TOF technology. The results herein are now included as a supplement to the P450 database.     

CYP2D6 and CYP2C19 genotypes and amitriptyline metabolite ratios in a series of medicolegal autopsies

In a series of 202 postmortem toxicology cases, the CYP2D6 and CYP2C19 genes were genotyped, and the concentrations of amitriptyline (AT) and six metabolites were analyzed. The polymorphic CYP2D6 and CYP2C19 genes encode enzymes participating in the metabolism of several potentially toxic drugs, and mutations in these genes may lead to adverse drug reactions, possibly even intoxications. AT was chosen as the substrate of interest because it is mainly metabolized by these enzymes, is considered relatively toxic, and ranks among the major causes of fatal drug poisoning in Finland. Our objective was to evaluate genetically determined interindividual variation in conjunction with metabolite ratios of drugs found in toxicological analysis in a series of medicolegal autopsies. Positive correlations were found between the proportion of trans-hydroxylated metabolites and the number of functional copies of CYP2D6 and between the proportion of demethylated metabolites and the number of functional copies of CYP2C19. None of the accidental or undetermined AT poisonings coincided with the CYP2D6 or CYP2C19 genotype which predicts a poor metabolizer phenotype. However, an unusually high femoral blood concentration of AT, 60mg/l, was found in one suicide case with no functional CYP2D6 genes. Our study shows a concordance of AT metabolite patterns with CYP2D6 and CYP2C19 genotypes in the presence of confounding factors typical for postmortem material. This result demonstrates the feasibility of postmortem pharmacogenetic analysis and supports the dominant role of genes in drug metabolism.     

A fatal doxepin poisoning associated with a defective CYP2D6 genotype

It has been suggested that the polymorphism of the CYP2D6 gene can contribute to occurrence of fatal adverse effects. We therefore investigated postmortem toxicology cases of fatal drug poisonings related to CYP2D6 substrates, with the manner of death denoted as accidental or undetermined. CYP2D6 genotypes were determined in 11 consecutive cases with samples available for DNA analysis. A case of fatal doxepin poisoning with an undetermined manner of death was found to coincide with a completely nonfunctional CYP2D6 genotype (*3/*4), indicating a total absence of CYP2D6 enzyme and suggesting a poor metabolizer phenotype. The doxepin concentration was 2.4 mg/L, the concentration of nordoxepin 2.9 mg/L, and the doxepin/nordoxepin ratio 0.83, the lowest found among the 35 nordoxepin-positive postmortem cases analyzed during the same year. No alcohols or other drugs were detected in the case. The CYP2C19 genotype was determined as that of an extensive metabolizer. The high N-desmethylmetabolite concentration is not consistent with acute intoxication. It is therefore probable that the defective genotype has contributed to the death, possibly involving repeated high dosage of doxepin. Our case strongly emphasizes that a pharmacogenetic analysis in postmortem forensic setting may reveal new insight to the cause or manner of death.     

评析“赵C案”

“赵C案”是一个典型的影响性诉讼案件，对公民维护自身姓名权具有标志性意义，此案过程比结果更加重要，因为它的教育意义更加深刻，也会让影响性诉讼更深入人心。     

INNOCENCE, ERROR, AND THE "NEW ABOLITIONISM": A COMMENTARY

If statistics are any indication, the system may well be allowing some innocent defendants to be executed.
- Justice Sandra Day O'Connor     

"A tempest in a cocktail glass": mothers, alcohol, and television, 1977-1996

This article examines the portrayal of pregnancy and alcohol in thirty-six national network evening news broadcasts (ABC, CBS, NBC). Early coverage focused on white, middle-class women, as scientific authorities and government officials warned against drinking during pregnancy. After 1987, however, women who drank during pregnancy were depicted as members of minority groups and as a danger to society. The thematic transition began before warning labels appeared on alcoholic beverages and gained strength from official government efforts to prevent fetal alcohol syndrome. The greatest impetus for the revised discourse, however, was the eruption of a "moral panic" over crack cocaine use. By linking fetal harm to substance abuse, the panic suggested it was in the public's interest to control the behavior of pregnant women.     

A Norms Approach to Jury "Nullification:" Interests, Values, and Scripts

Juries and other lay tribunals are often justified because they leaven the law with community norms. Unfortunately, we do not have a particularly good theory of when and how juries substitute their normative judgments for the law. A first step in developing such a theory is to examine the nature of norms and the way jurors bring normative judgments to their task. In this article I compare and contrast different understandings of norms that currently are in vogue in the social sciences and then use these approaches to develop a more systematic understanding of when juries do and when they do not substitute their normative judgment for that of the law.     

RESPONDING TO THE "CALL FOR PAPERS": Participation at A.S.C. Annual Meetings

The purpose of this study is to examine the participation of criminology and criminal justice graduate departments at the 1977-1979 American Society of Criminology meetings. Results show that institutional prestige. not productivity, is the leading predictor of meeting participation. In addition, meeting participation does not appear to be influenced by meeting site. Finally, it is suggested that criminologists focus more attention upon professional activities in order to facilitate our understanding of criminology as a science.     

“规范汉字”的法律意义——从“赵C”姓名权案到《现代汉语词典》第6版事件的法学联想

语言不仅是交流的工具,而且是文化身份的标志.语言规划立法,既有促进交流的目的,也担当着身份象征和文化传承的使命,应该综合这两方面的考虑,来理解语言规划立法所具有的特殊意义.我国公民享有姓名决定权,但仍应受到《国家通用语言文字法》和《居民身份证法》的限制,即公民居民身份证上的姓名必须使用规范汉字.因为我国目前在汉语词汇构成方面未有强制性标准,所以,《现代汉语词典》第6版收录“西文字母开头的词语”做法并没有“违法”.但是,面对字母词的使用现实及其可能引发的后果,我国确实应该尽快制定汉语词汇构成的国家标准,以明确西文缩略语的性质和词典编纂的规范要求.     

REVIEW ESSAY: DANGEROUS PEOPLE Criminal Careers and "Career Criminals" edited by Alfred Blumstein, Jacqueline Cohen, Jeffrey A. Roth, and Christy A. Visher. Washington, D.C.: National Academy Press, 1986.

Criminal Careers and "Career Criminals" edited by Alfred Blumstein, Jacqueline Cohen, Jeffrey A. Roth, and Christy A. Visher.     

The frequencies of mutated alleles of CYP2D6 gene in a Turkish population

Allele and genotype frequency distribution of CYP2D6*3, *4, *5, *6 and *10 variants were analyzed in blood samples of 100 unrelated healthy individuals by Real-Time PCR. The allele frequencies of CYP2D6*3(A2549del), *4(G1846A), *6(T1707del) and *10(C100T) were 1%, 10%, 2.5% and 14.5% respectively, while allele frequency of CYP2D6*5 was 3% of the subjects tested. Extensive, poor and intermediate metabolizer (EM, PM, IM) genotype frequencies were 63%, 4% and 12%, respectively. CYP2D6 gene duplication was 4%. Our results show that the frequencies of the mutated alleles of CYP2D6 in Turkish populations are similar to some European populations. 4% of Turkish people who have two nonfunctional defective allele are a high risk group and 12.5% of Turkish people who have two decreased functional defective allele or one normal and one non functional defective allele were also in the risk group. Findings of this study demonstrate the importance of genetic variation in drug intoxicants.     

法律信仰批判

中国法学界的"法律信仰"命题,是对美国学者伯尔曼有关法与信仰(宗教)之关系论述的误解,这种理论对中国社会影响很深,并导致了法律界的一系列错误观念。因此,有必要从理论上对这一命题进一步进行反思和批判。理论分析与实证研究表明,法律不能被信仰;我国建立法治的途径不能依赖法律信仰,而应是加强法与社会的沟通,增加法的现实性、可行性、合理性与正当性。     

批准(登记)生效合同、“申请义务”与“缔约过失” 《合同法解释(二)》第8条评注

本条规定了批准(登记)生效合同"成立后生效前"的"申请"义务以及违反该义务所生的责任,可作为请求权核心规范,与其他条文一起完成请求权规范链条,为此应在体系之中,按民法规范"要件——效果"的理念予以构造。在法教义学体系上,应重新审视本条与预约、条件、同意(追认)及违反强制性规定的关系。在适用范围上,一要注意基础行为与履行行为的区分,二要注意基础行为自身与前置行为的区分。在申请义务上,应作扩张解释,并灵活把握其主体及属性。在效果上,应综合考量批准要求之法规目的、信赖保护、机会损失等各种因素,妥当选择实际履行和损害赔偿等救济方式,厘定复杂的规范层次。尤须避免(违约责任进路下)缔约强制过于猛烈的影响。     

Development of a Tetraplex PCR Assay for CYP2D6 Genotyping in Degraded DNA Samples

CYP2D6 polymorphism analysis is gaining increasing interest in forensic pharmacogenetics. Nevertheless, DNA recovered from forensic samples could be of poor quality and not suitable for long polymerase chain reaction required to type CYP2D6 gene prior to SNaPshot minisequencing analysis performed to define alleles with different enzymatic activity. We developed and validated following the guidelines of the Scientific Working Group on DNA Analysis Methods a tetraplex PCR yielding four amplicons of 597, 803, 1142, and 1659 bp encompassing the entire CYP2D6 gene to analyze eleven SNP positions by SNaPshot minisequencing. Concordance, sensitivity, and specificity were assessed. The method, applied to thirty‐two forensic samples failed to amplify with long PCR, allowed the amplification of CYP2D6 gene in 62.5% of degraded samples. The new tetraplex PCR appears a suitable method for CYP2D6 analysis in forensic pharmacogenetics.