In this article, the genetic damage at 3 genetic endpoints in the workers occupationally exposed to antineoplastc drug was studied with micronucleus test, comet assay, hprt gene mutation assay and TCR gene mutation assay.

HCV的基因型及其变异与肝细胞癌的关系

丙型肝炎病毒(hepatitis CviruS,HCV)为单股正链RNA病毒.国内外已对数十个HCV全基因或片段基因进行了序列分析,并对HCV的基因型分型.近年研究表明,HCV的基因型及其变异与肝细胞肝癌(HCC)有者一定的关系.其中,HCV1b基因型与HCC的发生显著相关;HCV核心区的变异与HCC的发生可能相关;HCV NS3与HCC的发生相关;HCV NS5的序列相对稳定为诱发HCC所必须的;而HCV E2/NS1与HCC的发生尚无定论.丙型肝炎及由其发展而来的肝细胞癌至今没有令人满意的治疗方法.研究HCV基因型及其变异与HCC发生的关系对阐明HCC发生的机理和HCC的防治有着重要的意义,为HCC的治疗开辟了新的途径,成为近年来国内外研究的热点,通过对他的深入研究可能为HCC的基因治疗找到有效措施.

胃癌癌前和发展阶段中E-cadhherin基因的突变

目的:细胞粘附分子E-cadherin作为转移抑制因子在肿瘤发展过程中起者重要的作用,在弥漫型家族性胃癌中有E-cadherin基因胚系突变的报道,并且在弥漫型散发性胃癌中也有关于该基因失活性突变的报道.异型增生是一种常见的胃癌癌前病变,为了认识E-cadherin基因在散发性胃癌发展过程中的分子改变,我们对不同地区不同阶段的胃组织标本进行了E-cadherin基因突变的检测.方法:采用变性梯度凝胶电泳(DGGE)和测序的方法对130例癌前、早癌及进展期胃癌组织进行了第6、7、8、9外显子的突变检测,其中高发区早癌40例,非高发区早癌30例,异型增生40例,进展期胃癌20例.结果:在一例进展期胃癌中检测到第9外显子的错义突变,该突变是文献未报道过的新突变,在早期胃癌和异型增生中都未发现突变.结论:E-cadherin基因突变在散发胃癌为少发事件,不是胃癌发生早期阶段的主要事件,在胃癌发生的早期阶段可能有其他更重要的机制.

We previously found that the K141N mutation in heat shock protein B8 (HSPB8) was respon-sible for Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of the present study was to generate a transgenic mouse model bearing the K141N mutation in the human HSPB8 gene, and to determine whether this K141NHSPB8 transgenic mouse model would manifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L, and consequently be suitable for use in studies of disease pathogenesis. Transgenic mice overexpressing K141NHSPB8 were generated using K141N mutant HSPB8 cDNA cloned into a pCAGGS plasmid driven by a human cytomegalovirus expression system. PCR and western blot analysis conifrmed integra-tion of the K141NHSPB8 gene and widespread expression in tissues of the transgenic mice. The K141NHSPB8 transgenic mice exhibited decreased muscle strength in the hind limbs and impaired motor coordination, but no obvious sensory disturbance at 6 months of age by behavioral assess-ment. Electrophysiological analysis showed that the compound motor action potential amplitude in the sciatic nerve was signiifcantly decreased, but motor nerve conduction velocity remained normal at 6 months of age. Pathological analysis of the sciatic nerve showed reduced myelinated ifber density, notable axonal edema and vacuolar degeneration in K141NHSPB8 transgenic mice, suggesting axonal involvement in the peripheral nerve damage in these animals. These ifndings indicate that the K141NHSPB8 transgenic mouse successfully models Charcot-Marie-Tooth disease type 2L and can be used to study the pathogenesis of the disease.

OBJECTIVE: To evaluate the association of X-ray cross-complementing group 1 (XRCC1) Arg399Gln, Arg194Trp and Arg280His polymorphisms with the risk of glioma. DATA SOURCES:A systematic literature search of papers published from January 2000 to August 2012 in PubMed, Embase, China National Knowledge Infrastructure database, and Wanfang da-tabase was performed. The key words used were“glioma”,“polymorphism”, and“XRCC1 or X-ray repair cross-complementing group 1”. References cited in the retrieved articles were screened manual y to identify additional eligible studies. STUDY SELECTION: Studies were identified according to the fol owing inclusion criteria:case-control design was based on unrelated individuals;and genotype frequency was available to estimate an odds ratio (OR) and 95%confidence interval (CI). Meta-analysis was performed for the selected studies after strict screening. Dominant and recessive genetic models were used and the relationship between homozygous mutant genotype frequencies and mutant gene frequency and glioma incidence was investigated. We chose the fixed or random effect model according to the heterogeneity to calculate OR and 95%CI, and sensitivity analyses were conducted. Publication bias was examined using the inverted funnel plot and the Egger’s test using Stata 12.0 software. MAIN OUTCOME MEASURES: Association of XRCC1 Arg399Gln, Arg194Trp, and Arg280His polymorphisms with the risk of glioma, and subgroup analyses were performed according to differ-ent ethnicities of the subjects.
RESULTS:Twelve articles were included in the meta-analysis. Eleven of the articles were concerned with the Arg399Gln polymorphism and glioma onset risk. Significantly increased glioma risks were found only in the dominant model (Gln/Gln+Gln/Arg versus Arg/Arg:OR=1.26, 95%CI=1.03-1.54, P=0.02). In the subgroup analysis by ethnicity, significantly increased risk was found in Asian subjects in the recessive (OR = 1.46, 95%CI = 1.04-2.45, P = 0.03) and dominant models (OR = 1.40, 95%CI = 1.10-1.78, P =0.007), and homozygote contrast (OR=1.69, 95%CI=1.17-2.45, P=0.005), but not in Caucasian sub-jects. For association of the Arg194Trp (eight studies) and Arg280His (four studies) polymorphisms with glioma risk, the meta-analysis did not reveal a significant effect in the al ele contrast, the recessive genetic model, the dominant genetic model, or homozygote contrast. CONCLUSION: The XRCC1 Arg399Gln polymorphism may be a biomarker of glioma susceptibility, es-pecial y in Asian populations. The Arg194Trp and Arg280His polymorphisms were not associated with overal glioma risk.

建立PCR-高分辨熔解分析法检测急性髓系白血病IDH1基因突变

可溶性异柠檬酸脱氢酶1(IDH1)基因在细胞氧化损伤反应中发挥着重要的调控作用[1].2008年Parsons等[2]首先报道胶质瘤患者中12%IDH1转录本第395位碱基由鸟嘌呤突变为腺嘌呤(c.395G→A),导致该酶精氨酸被组氨酸所替代(R132H).其后发现IDH1基因突变还可见于星形细胞瘤、少突神经胶质瘤、急性髓系白血病(AML)、骨髓增生异常综合征、骨髓增殖性肿瘤等[3].高分辨率熔解(HRM)分析是近年来发展起来的一种新型的突变扫描和基因分型的遗传学分析方法.美国Idaho公司的LightScanner是以HRM技术为基础的分析仪,它采用新型饱和双链DNA结合饱和荧光染料LC Green,通过分析PCR扩增产物解链过程中熔解曲线的变化来检测单碱基突变、小片段插入或缺失[4],具有灵敏度高、特异性好、成本低廉、高通量检测的优点,已越来越多地被用于单核苷酸多态性分析(SNP)和基因突变扫描.

先天性并(多)指(趾)畸形家系的HOXD13基因突变分析

先天性并(多)指(趾)(synpolydactyly,SPD)为常染色体的显性遗传病,常有家族发病的倾向.SPD是目前发现的第一个由HOXD13突变所导致的人类畸形综合征.

一先天性厚甲家系角蛋白基因突变鉴定

目的 探讨一个先天性厚甲家系角蛋白基因突变.方法 用PCR及Sanger测序技术对先天性厚甲家系先症者KRT17基因所有外显子和KRT6B基因编码螺旋起始和终止区域序列进行突变鉴定,针对发现的可疑位点,Sanger测序检测家系其他成员该位点的变异情况.结果 基因检测结果表明,家系患者KRT17基因错义突变c.263T＞C,该突变导致角蛋白17(K17)第88位氨基酸由蛋氨酸变成苏氨酸(p.M88T),KRT6B基因未见异常.结论 KRT17基因c.263 T＞C(p.M88T)突变是该先天性厚甲家系致病基因突变.

甘露糖结合蛋白基因突变与感染性疾病间关系的研究进展

甘露糖结合蛋白(mannose binding protein,MBP)或甘露糖结合凝集素(mannose binding lectin,MBL),是一种钙依赖性糖结合蛋白,属于凝集素家族,可与甘露糖残基结合,参与细胞表面识别,广泛存在于人的肝脏和血液中,是天然免疫(innate immunity)的重要组成部分.MBP的功能及基因结构和感染性疾病的关系近年来倍受关注.

毛细管电泳PCR-SSCP分析p16基因突变

Lung cancer is one of the most common cancers in the world. Some genetic alterations such as p53 gene and ras gene mutations, have been identified in this disease. Recently, a putative tumor suppressor gene, the p16/CDKN2/MTS1 gene containing 3 extrons and 2 introns, located in the chromosome p21 region, was cloned independently by three research groups. Traditionally, gene mutation analysis was performed by slab polyacrylamide gel electrophoresis. However, this method is laborious, time-consuming, low sensitivity and harmful to human health. Capillary electrophoresis (CE) with the characteristics of rapidity and high performance has numerous advantages over conventional slab polyacrylamide gel electrophoresis. An important advantage of CE is that the commercially available system is automation.

Objective:To investigate the role of Mdm2 expression in hepatocellular carcinoma.Methods: Streptavidin-peroxidase conjugation method (SP)was used to observe the expression of Mdm2 and p53 proteins in 61 cases of hepatocellular carcinoma(HCC)and 59 cases of corresponding paracancerous tissues,among which p53 mutations in exons 5～8 were detected in 21 cases by PCR-SSCP.Results:Positive nuclear P53 and Mdm2 immunostains were demonstrated in 57.38％ and 26.23％ of HCC,1.69％ and 3.39％ of corresponding paracancerous tissues respectively.The expression of p53 and Mdm2 proteins in hepatocellular carcinoma was significantly higher than that in paracancerous tissues(P＜0.01).The expression of P53 and Mdm2 was not significantly correlated.42.86％ of hepatocellualr carcinomas showed mutations in exon 7 of p53 gene,and no mutation was found in exons 5,6,8 and paracancerous tissues. 66.67％ of mutational cases had P53 overexpression and 11.11％(1/9)showed overexpression of both P53 and Mdm2,Mdm2 overexpression also appeared in 25％ of cases without mutations.Conclusions:Mdm2-induced p53 inactivation and p53 gene mutation play an important role in carcinogenesis of hepatocellular carcinoma.Tumorigenic property of Mdm2 itself,together with p53 mutation,may take part in hepatocarcinogenesis.

肺癌EGFR基因突变检测方法的研究进展

表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变与肺癌病人对抗肿瘤药物-吉非替尼的敏感性有关[1,2].而肺癌的前期临床试验[3,4]表明,在含铂类药物联合化疗失败后,吉非替尼仍有12%-18%的客观有效率,疾病控制率在50%左右,中位生存期约为7个月左右,受试者的症状改善且安全性良好,所以EGFR基因突变检测对于临床医生进行肺癌的个体化治疗具有重要的参考价值.关于EGFR基因突变检测的方法,文献报道主要来源于国外,但当前仍然没有一种完全适用于临床的方法.现就肺癌EGFR基因突变检测方法的研究作一综述.

小睑裂综合征的基因定位和突变研究进展

小睑裂综合征又称睑裂狭小、倒转型内眦赘皮和上睑下垂综合征(blepharophimosis epicanthus in-versus and ptosis, BPES),1875年,Galeyowski首先报告本病,

Background: Dent's disease is a rare X-linked recessive hereditary disease caused by mutations in either the CLCN5 or OCRL1 genes. This disease is characterized by manifestations of proximal renal tubule dysfunction associated with low molecular weight proteinuria (LMWP), hypercalciuria, nephrocalcinosis, nephrolithiasis, and progressive renal failure.
Methods: We report a Chinese boy with Dent's disease, clinically diagnosed by LMWP and hypercalciuria. Genetic analysis was made of the CLCN5 and OCRL1 genes. Related studies were also reviewed.
Results: A splice site mutation IVS6, +2T>C of the CLCN5 gene was revealed in this case, and it was not reported previously.
Conclusions: Clinical and genetic analysis is valuable for the diagnosis of Dent's disease. A novel mutation in the CLCN5 gene was identifi ed in our patient.