韩国人CYP450 1A1、CYP450 2E1和GSTM1、GSTT1、GSTP1基因座遗传多态性分析

目的 调查代谢相关的CYP4501A1、CYP4502E1和GSTM1、GSIT1、GSTP1基因座在韩国人群中的遗传多态性分布状况。方法 采用多重聚合酶链式反应、聚合酶链式反应-限制性片段长度多态性技术，分析300名韩国健康大学生的CYP1A1基因3′端限制性内切酶Msp Ⅰ位点、CYP2E1基因5′端转录调节区Pst Ⅰ位点和GSTM1、GSTT1缺失与存在、GSTP1基因第5外显子BsmA Ⅰ位点的基因型，计算基因型和基因频率。结果 CYP1A1基因型频率为ml／ml型39．7％、ml／m2型49．7％、m2／m2型10．7％，基因频率为ml 0．645、m2 0．355。CYP2E1基因型频率为cl／cl型66．7％、cl／c2型30％、c2／c2型3．3％，基因频率为C1 0．818、C2 0．182。GSTM1基因缺失型频率为53．3％。GSTT1基因缺失型频率为54．7％。GSTP1基因型频率为Ile／Ile型62％、Ile／Val型34．3％、VaL／Val型3．7％，基因频率为Ile 0．792、Val 0．208。基因分布符合Hardy-Weirtberg平衡定律。结论 韩国人CYP1A1、CYP2E1、GSTM1、GSTT1基因分布与我国人群较为相近，半数以上人缺乏GSTM1和GSTT1基因，纯合缺失型频率超过印度人的3倍。     

DMD基因突变及突变检测技术研究进展

假性肥大型进行性肌营养不良（DMD／BMD）一类发病率较高的X连锁隐性遗传病。DMD基因庞大,突变机制复杂,随着分子生物学的进展,对DMD基因及其所编码aystrophin蛋白的认识不断深入,探索更简便、准确、经济的基因突变检测技术成为目前对DMD研究的热点,本文着重对DMD基因、突变与表型关系的研究进展,以及突变检测技术的进展做一综述。     

ER、PR、P53、P16和CerbB-2在乳腺癌中的表达及与临床、病理的关系

目的联合检测抑癌基因P53、P16和癌基因CerbB-2在乳腺癌中的表达情况,结合ER、PR探讨与乳腺癌发生发展及临床、病理因素之间的联系.方法采用免疫组化S-P法分别检测145例乳腺癌和10例正常乳腺组织ER、PR、P53、P16和CerbB-2蛋白的表达.结果145例乳腺癌中ER、PR、P53、P16和CerbB-2的阳性表达率分别为43.4%、40.6%、45.5%、27.5%和84.8%.与正常乳腺组织比较,P53、P16和CerbB-2的阳性表达有差异（p＜0.05）.乳腺癌各病理类型之间表达没有差异（p＞0.05）.ER与年龄、PR、CerbB-2、肿瘤直径、月经状态、癌周浸润相关（p＜0.05）;PR与ER、肿块直径、癌周浸润相关;P53与淋巴结转移个数,癌周浸润相关;P16与初潮年龄、癌周浸润相关（＜0.05）;CerbB-2与ER、肿瘤直径、转移淋巴结个数、月经状态、癌周浸润相关（p＜0.05）.结论ER、PR是对激素治疗有价值的指标,并与P53、P16、CerbB-2等可作为评价乳腺癌的生物学行为、病理特征和预后的指标.     

A report of the first biennial meeting on Capita Selecta in Complex Disease Analysis (CSCDA2010), Leuven, Belgium, August 25-27, 2010

There is a need for interdisciplinary assessments and interpretations of -omics underpinnings of human complex diseases. However, often investigators from different, yet overlapping, disciplines experience difficulties in understanding the other discipline's language and there is a clear need for establishing a platform that nourishes interdisciplinary team processes and allows tearing down the professional's tower of Babel. To accommodate these needs, the biennial mini-conference Capita Selecta in Complex Disease Analysis was instigated. Abstracts are freely available online [http://www.aimontefiore.org/cscda2010/].     

癌基因PPO及其同源性基因的结构分析

目的采用PPO(proliferation and phosphorylation oncogene)基因与其同源性基因的结构分析,推测PPO并验证其功能。方法利用PPO的蛋白序列寻找同源基因,比较其结构并利用蛋白免疫杂交方法证实其功能。结果克隆了PPO基因,找到了PPO基因与小鼠、果蝇、蚊子、线虫以及酵母等的同源基因,比较发现PPO在进化上非常保守。人和小鼠的氨基酸序列有许多与磷酸化有关的功能域,并发现其中某些氨基酸在进化上非常保守。进一步研究表明PPO能使ERK2和MEK磷酸化。结论PPO基因在进化上非常保守,与磷酸化功能有关。     

Brazilian P[8],G1, P[8],G5, P[8],G9, and P[4],G2 rotavirus strains: nucleotide sequence and phylogenetic analysis

Rotavirus epidemiological surveys with molecular analysis of strains are required for gastroenteritis control and prevention. Twenty-nine human rotavirus strains detected in Rio de Janeiro, Brazil, from 1986 to 2004 were characterized as P[8],G1, P[8],G5, P[8],G9, and P[4],G2 genotypes. The VP7 genes were sequenced and phylogenetic analysis was performed. Strains of genotype G1 revealed two distinct lineages, G1-3 and G1-4; strains of genotype G2 grouped in lineage G2-1; G5 strains clustered with other Brazilians G5 strains and G9 strains were closely related to each other in lineage G9-3, distinct from the original G9 strains detected in 1980s. The VP4 genes were analyzed and P[8] strains fell into two major genetic lineages, P[8]-2 and P[8]-3. Our findings document an intragenotype diversity represented by lineages and sublineages within rotavirus circulating in Rio de Janeiro from 1986 to 2004, before application of a vaccine (Rotarix) in Brazil. This report emphasizes the importance of continuing monitor genotypes to verify if uncommon strains or newly strains are emerging to be specifically addressed in future vaccine trials.     

Genetic variation in biotransformation enzymes,air pollution exposures,and risk of spina bifida

Spina bifida is a birth defect characterized by incomplete closure of the embryonic neural tube. Genetic factors as well as environmental factors have been observed to influence risks for spina bifida. Few studies have investigated possible gene‐environment interactions that could contribute to spina bifida risk. The aim of this study is to examine the interaction between gene variants in biotransformation enzyme pathways and ambient air pollution exposures and risk of spina bifida. We evaluated the role of air pollution exposure during pregnancy and gene variants of biotransformation enzymes from bloodspots and buccal cells in a California population‐based case‐control (86 cases of spina bifida and 208 non‐malformed controls) study. We considered race/ethnicity and folic acid vitamin use as potential effect modifiers and adjusted for those factors and smoking. We observed gene‐environment interactions between each of the five pollutants and several gene variants: NO (ABCC2), NO₂ (ABCC2, SLC01B1), PM₁₀ (ABCC2, CYP1A1, CYP2B6, CYP2C19, CYP2D6, NAT2, SLC01B1, SLC01B3), PM_2.5 (CYP1A1 and CYP1A2). These analyses show positive interactions between air pollution exposure during early pregnancy and gene variants associated with metabolizing enzymes. These exploratory results suggest that some individuals based on their genetic background may be more susceptible to the adverse effects of pollution.     

Trinucleotide repeat polymorphism within exon 5 of the MICA gene (MHC class I chain-related gene A): allele frequency data in the nine population groups Japanese, Northern Han, Hui, Uygur, Kazakhstan, Iranian, Saudi Arabian, Greek and Italian

We recently identified a trinucleotide repeat polymorphism, (GCT)n, within the transmembrane (TM) segment of the human MHC class I MICA gene (MHC class I chain-related gene A). Five distinct alleles (A4, A5, A5.1, A6, A9) corresponding to 4, 5, 5 with one nucleotide insertion, 6 and 9 repetitions, respectively, have been detected in various HLA-homozygous B cell lines. Here we present allele frequencies for this trimeric short tandem repeat (STR) in 604 unrelated individuals collected from nine human populations (Japanese, Northern Han, Hui, Uygur, Kazakhstan, Iranian, Saudi Arabian, Greek and Italian) determined using the polymerase chain reaction (PCR) combined with fluorescent-based automated fragment detection technology. All alleles were present in each population, but allelic distributions varied from one population to another. No new alleles (such as A7 or A8) were identified. The evolutionary and structural significance of these data as well as the potential application to forensic medicine is discussed.     

Reliability of the ica, aap and atlE genes in the discrimination between invasive, colonizing and contaminant Staphylococcus epidermidis isolates in the diagnosis of catheter-related infections

Objectives   To evaluate the usefulness of detecting two genes involved in biofilm formation ( icaA and aap ) and one gene involved in initial adhesion ( atlE ) for discrimination between contaminant, colonizing and invasive Staphylococcus epidermidis isolates involved in catheter-related infections.
Patients   The first group contained 29 isolates that were isolated from the skin of healthy volunteers (contaminant isolates). The second group contained 16 isolates recovered from catheters (>1000 CFUs on quantitative catheter culture) from asymptomatic patients without bacteremia. These isolates were considered to be colonizing isolates. The third group contained 34 isolates grown in ≥2 different blood cultures from patients with a systemic inflammatory response. These isolates were considered to be invasive isolates.
Results   The prevalence of atlE did not differ between the three groups. The icaA and aap genes were significantly more prevalent in colonizing isolates (88% aap ; 88% icaA ) than in invasive isolates (68% aap , P  = 0.179; 59% icaA , P  = 0.055) and than in skin isolates (52% aap , P  = 0.02; 38% icaA , P  = 0.002).
Conclusions   The high prevalence of aap and icaA in skin isolates and their higher prevalence in colonizing than in invasive isolates led to a low specificity when these genes were used to differentiate between contamination, colonization and invasive infection. We conclude that, although the prevalence of these genes differs in the three groups, their presence cannot be used for clinical decision-making.     

Livin基因、FHIT基因与妇科肿瘤间关系研究进展

目的:总结Livin基因、FHIT基因与妇科肿瘤的关系.方法:在Pubmed、中国知网查阅有关Livin基因、FHIT基因与妇科肿瘤关系的文献,进行汇总分析.结果:在妇科肿瘤中,Livin基因高表达、FHIT基因低表达,抑制细胞凋亡,促进肿瘤的发生.结论:通过对Livin基因、FHIT基因的深入研究,为妇科肿瘤的治疗提供新靶点.     

染色体9p21-22区域两个新的SNPs位点在湖南汉族人群中的多态分布

目的 在人类染色体9p21—22区域克隆的两个候选抑瘤基因NGX6和UBAPl以及该区域另一个已知抑瘤基因CDKN2A中筛查未知的单核苷酸多态(single nucleotide polymorphisms，SNPs)。方法 用96名正常人外周血白细胞基因组DNA在3个基因的外显子及基因上下游调控区中逐段PCR扩增、大规模涵序。结果 在UBAP1基因第6外显子及CDKN2A基因第4外显子中各发现一个新的SNP位点，dbSNP登录号为rs3135929和rs3088440，其多态信息含量分别为0．102和0．213。两个SNPs之间不存在连锁不平衡，单倍型分析表明两个SNPs联用，其多态信息含量可达到0．302。结论多 个SNP联用，其多态信息含量明显提高，可克服SNPs信息量不足的弱点；两个SNPs均位于基因的3'非翻译区，对基因表达调控是否有影响值得进一步研究。     

人Hes1-shRNA,Hes5-shRNA慢病毒表达载体的构建、包装及鉴定

目的 构建人Hes1-shRNA和Hes5-shRNA慢病毒表达载体,为Notch-Hes信号通路的相关研究奠定基础.方法 根据人Hes1,Hes5基因mRNA序列分别设计、合成多对互补的DNA单链寡核苷酸,退火后克隆至pENTR /U6入门载体.通过入门载体瞬时转染神经胶质瘤U251细胞筛选有效干扰序列.将含有效干扰序列的入门载体与pLenti6/BLOCK-iT-DEST载体进行LR重组构建Hes1-shRNA和Hes5-shRNA慢病毒表达载体,经脂质体介导入293FT细胞,包装成慢病毒.用该慢病毒感染U251细胞,Western印迹法分别检测Hes1,Hes5蛋白的表达.结果 分别构建了针对Hes1和Hes5基因的特异性shRNA慢病毒表达载体,其包装获得慢病毒可有效感染U251细胞并分别对Hes1,Hes5蛋白的表达有显著抑制作用.结论 成功构建了Hes1-shRNA和Hes5-shRNA慢病毒表达载体.     

Genes,factor X,and allergens: what causes allergic diseases?

Genetic disposition and allergen exposure play the main roles in the development of allergic diseases. Another factor that could be involved is the nutritional intake of vitamin D.     

结直肠癌患者肿瘤组织中KRAS、NRAS和BRAF基因突变的分子病理检测分析

目的：探讨结直肠癌患者肿瘤组织中KRAS、NRAS和BRAF基因各亚型突变状况。方法：应用Taqman-ARMS方法检测101例结直肠癌患者石蜡组织中KRAS、NRAS、BRAF基因突变情况。结果：结直肠癌患者肿瘤组织中KRAS基因总突变率为42.57%,其中30例(29.70%)检测到外显子2第12密码子突变;其中13例(12.87%)检测到KRAS基因外显子2第13密码子突变,为G13D点突变,未检测到G13C点突变;NRAS基因总突变率为3.96%,其中外显子2第12密码子突变1例(0.99%),外显子2第13密码子突变1例(0.99%),外显子3第61密码子突变2例(1.98%);BRAF基因总突变率为3.96%。结论：结直肠癌患者组织中KRAS基因突变率较高,NRAS和BRAF基因突变率虽低但不容忽视。KRAS、NRAS和BRAF基因检测人群的筛选对结直肠癌患者治疗方案的选择意义重大,能够更有效的指导精准医学个体化治疗。     

Genetic changes of CDH1, APC, and CTNNB1 found in human brain tumors

This paper focuses on changes in E-cadherin (CDH1), adenomatous polyposis coli (APC), and beta-catenin (CTNNB1) in 50 tumors of the central nervous system. All gene products are components of adherens junctions, but are also involved in wnt signaling. The results of our analysis showed LOH of CDH1 gene in 31% of meningiomas examined (significant correlation; p=0.002). LOH was noted in a single case of germinoma, while other tumor types did not demonstrate any change in CDH1. Fourteen samples (29.2%) with changes in APC gene were observed. The changes were seen in 33.3% of glioblastomas and in 27% of meningiomas; LOH occurred in five informative astocytomas (20%) and in six informative neurinomas (17%). One oligoastrocytoma showed LOH at exon 11, and one medulloblastoma had allelic imbalance at both exons. Five samples (10%) showed heteroduplexes in exon 3 of beta-catenin. Potential mutations were confined to two meningiomas, one astrocytoma, one glioblastoma, and one germinoma. Our results suggest that genetic changes in wnt components are involved in brain tumor genesis. Changes in E-cadherin are involved in meningiomas, while changes in APC gene occur in different tumor types, with glioblastomas showing the highest percentage.     

Sequence analysis of the capsid gene of Aichi viruses detected from Japan, Bangladesh, Thailand, and Vietnam

Sequence analysis of the capsid gene of Aichi viruses was performed on 12 strains detected in Japan, Bangladesh, Thailand, and Vietnam during 2002-2005. The phylogenetic tree constructed from 17 nucleotide sequences of the capsid gene of the strains studied and reference strains demonstrated that Aichi virus strains clustered into two branches. A classification of Aichi viruses based on the capsid gene was proposed, in which lineage I consists of the Aichi virus strains detected from Japan, Thailand, Vietnam, and Germany, and lineage II includes Bangladeshi strains and a Brazilian strain.