N-乙酰基转移酶基因多态性与帕金森病相关性研究

目的　探讨N -乙酰基转移酶 (NAT2 )基因多态性与散发性帕金森病 (Parkinson’sdisease ,PD)的关系。方法　应用自动实时荧光Light-Cycler技术 ,分析 88例PD患者和 112例健康人NAT2 4个位点的基因多态性 ,比较PD患者与对照组间频率差异。结果　早发PD组NAT2 6A等位基因频率与对照组比较有显著性差异 (P <0 .0 5 ) ,使患PD的危险度提高了 2 .0 8倍 (P <0 .0 5 ) ,NAT2 5A和NAT2 7A/B等位基因频率与对照组比较无显著性差异 (P >0 .0 5 ) ;晚发PD组NAT2 4个多态位点各等位基因频率与对照组比较无显著性差异 (P >0 .0 5 ) ;未检测到NAT2 14A等位基因。结论　NAT2 6A等位基因可能主要与早发PD的易感性相关 ,并参与了神经毒素的解毒。     

子宫内膜异位症与细胞色素P450 1A1基因A4889G突变的相关性

目的探讨细胞色素P450 1A1(cytochrome P450 1A1,CYP1A1)基因A4889G突变与子宫内膜异位症(endometriosis,EM)的遗传易感性的关系.方法采用等位基因特异性聚合酶链反应的方法,研究了76例EM患者和80名正常对照CYP1A1基因A4889G位点碱基突变.两组均为广东籍汉族妇女.结果 CYP1A1基因A4889G位点等位基因A、G在EM组和对照组分布的差异有显著性(χ2=7.498,P<0.01),其中等位基因G使患EM的相对危险度提高了1.957倍.CYP1A1基因A4889G位点各基因型分布在两组间差异有显著性(χ2=6.915,P<0.05),GG基因型与AA基因型相比患EM的危险度高3.437倍(χ2=5.430,P<0.05).结论 CYP1A1基因A4889G突变等位基因与EM有一定的关联,突变基因型增加了EM的发病风险.     

中国北方汉族人细胞色素P450 1A1基因MspⅠ多态性与早发性帕金森病关系的研究

目的：探讨中国北方汉族人细胞色素P4501A1基因MspI多态性与早发性帕金森病的易感性的关系。方法：用聚合酶链反应－限制性片段长度多态性技术,分析了126例早发性帕金森病患者（发病年龄＜50岁）和172名正常健康成人CYP1A1基因3’端限制性内切酶MspI位点的3种基因型（A、B、C）的分布频率。结果：MspI基因型C在病例组和对照组中各占15．1％和13．4％,基因型A在两组中分别占41．3％和34．9％,基因型B在两组中分别占43．6％和51．7％,各基因型在两组中相比较,差异无显著性（P＞0．05）。C基因型在病例组和对照组分别占36．9％和39．2％,两者差异无显著性（P＞0．05）。结论：提示解毒酶CYP1A1基因MspI多态性的单独存在可能与早发性帕金森病的易患性无关。     

CYP1A1、CYP2D6基因多态性对白血病发生的影响

目的 分析细胞色素P450 CYP1A1和CYP2D6的多态性基因型在湖南地区白血病患者和健康人群中的分布及其对白血病发生的影响.方法 采用PCR及PCR-RFLP技术分析多态性基因型频率.结果 CYP1A1和CYP2D6基因的野生型、杂合突变型及纯合突变型的分布频率在急性淋巴细胞性白血病、急性非淋巴细胞性白血病、慢性粒细胞性白血病患者组与健康对照组之间无显著性差异;携带一个突变等位基因型的个体患白血病的风险与相应野生型携带者比较均无显著性差异;急性非淋巴细胞性白血病患者组的CYP1A1杂合突变型与CYP2D6杂合突变型的联合基因型频率高于健康对照组.结论 单独的CYP1A1或CYP2D6基因的多态性变异与白血病易感性不相关;CYP1A1杂合突变与CYP2D6杂合突变的联合基因型增加患急性非淋巴细胞性白血病的风险.     

CYP1B1基因多态性与子宫内膜异位症易感性的相关研究

目的 研究CYP1B1基因第2外显子119(G-T)、第3外显子432(C-G)多态性与子宫内膜异位症(endometriosis,Ems)易感性的关系.方法 采用等位基因特异性聚合酶链反应对55例Ems患者和45例对照组进行CYP1B1基因第2外显子119(G-T)、第3外显子432(C-G)突变分析,探讨Ems的发生与CYP1B1基因多态性之间的相关性.结果 CYP1B1基因密码子119中等位基因G、T在Ems组和对照组分布的差异有统计学意义(P＜0.05),其中等位基因T使Ems发病风险提高2.061倍;CYP1B1基因密码子119G/T各基因型分布两组间差异有统计学意义(P＜0.05),纯合突变(T/T)基因型、杂合突变(G/T)基因型与野生型(G/G)基因型相比,患Ems的危险度分别为2.625倍和3.214倍.以CYP1B1联合野生型GG和CC个体的OR值为1相比,CYP1B基因密码子119杂合型突变(Ala/Ser)合并密码子432野生型个体的OR值为2.976,95%CI:1.129～7.848,P＜0.05.结论 CYP1B1基因第2外显子119(G-T)突变等位基因与Ems的发生有一定关系,突变基因型增加了Ems的发病风险;CYP1B1基因第2外显子杂合型突变(Ala/Ser)联合密码子432野生型能增加Ems的发病风险.     

CYP1A1基因I462V多态与非小细胞肺癌的相关性

目的 探讨CYP1A1基因1462V多态性与非小细胞肺癌的相关性.方法 采用病例对照研究,应用聚合酶链反应-限制性片段长度多态性对72例非小细胞肺癌患者(病例组)和90例正常对照(对照组)CYP1A1基因I462V多态进行检测,分析基因型频率和等位基因频率在病例组和对照组的分布,比较不同基因型与非小细胞肺癌患病风险的关...     

中国北方汉族人细胞色素P4501 A1基因MspⅠ多态性的研究

目的　探讨中国北方汉族人细胞色素P(cytochromeP ,CYP) 4 5 0 1A1基因MspⅠ多态性。方法　用聚合酶链反应 限制性片段长度多态性 (PCR RFLP)技术 ,分析了 172名北方汉族正常健康成人CYP1A1基因 3′端限制性内切酶MspⅠ位点的3种基因型 (A、B、C)的分布频率。结果　MspⅠ等位基因m1、m2分别占 6 0 8%、39 2 %。MspⅠ基因型A占 34 9% ,基因型B占 5 1 7% ,基因型C占 13 4 %。结论　本研究结果提示中国北方汉族人解毒酶CYP1A1基因存在MspⅠ多态性。     

中国人NAD(P)H:醌氧化还原酶基因多态性与帕金森病遗传易感性的关系

目的　探讨依赖还原型辅酶 / 醌氧化还原酶 [NAD(P) H:quinone oxidoreductase,NQO1]c DNA6 0 9位点 C→ T多态性与帕金森病 (Parkinson'sdisease,PD)遗传易感性的关系。方法　用聚合酶链反应 -变性高效液相技术 (polymerase chain reaction- denaturing high performance liquid chromatog-raphy,PCR- DHPL C)分析了 NQO1基因c DNA6 0 9位点C→T多态性在 PD患者与正常对照之间分布频率的差异。结果　PD组和对照组的 TT基因型频率分别为 2 2 .6 %和 11.8% (P=0 .0 0 4 ) ,TT基因型使患 PD的危险度提高 2 .186倍 (P=0 .0 0 5 ) ;根据发病年龄分组后 ,这种差异主要存在于晚发性 PD和对照组之间 ,TT基因型使患 PD的危险度提高 2 .6 2 7倍 (P=0 .0 0 1)。等位基因在总体 PD组、早发 PD组、晚发 PD组和对照组中的频率分布差异无显著性。结论　NQO1基因c DNA6 0 9位点C→T多态性在对照组和PD患者之间的分布差异有显著性 ,突变基因型 (TT基因型 )频率在 PD组中较高 ,研究结果支持 NQO1基因多态性与 PD相关的假说 ,而且与 PD发病年龄有关。     

APOA5基因单核苷酸多态性与冠心病相关性研究

目的　探讨APOA5基因多态性与冠状动脉粥样硬化性心脏病 (coronaryheartdisease ,CHD)的相关性、与血脂关系及其在中国汉族人群中的分布。方法　用聚合酶链反应 限制性片段长度多态性分析APOA5与APOA4交界区域的T/C单核苷酸多态。结果　T/C单核苷酸多态位点等位基因T、C频率在CHD组和正常对照组分别为 0 .43 5、0 .5 65和 0 .3 74、0 .62 6。等位基因频率和基因型频率分布均符合Hardy Weinberg平衡定律。T/C基因多态性基因型频率 ,等位基因T、C频率在两组间差异有显著性(P <0 .0 5 ) ,且基因型CC的冠心病患者其血浆高密度脂蛋白水平显著高于其他基因型患者 (P <0 .0 1)。中国人T/C单核苷酸多态位点T、C等位基因频率与欧洲白人比较 ,差异存在非常显著性 ( 0 .3 74vs 0 .663、0 62 6vs 0 .3 3 7;P <0 .0 0 1)。结论　T/C单核苷酸多态性与CHD存在相关性 (P <0 .0 5 ) ,患者组CC基因型与血浆高密度脂蛋白水平密切相关 (P <0 .0 1)。     

细胞代谢解毒酶CYP1A1、GSTM1基因多态性与吸烟对男性肺鳞癌发病的影响

 目的 探讨细胞代谢解毒酶细胞色素P450酶1A1（CYP1A1）、谷胱甘肽S-转硫酶M1（GSTM1）基因多态性和吸烟因素对男性肺鳞癌发病的影响。方法 采用基因芯片技术对125例男性肺鳞癌患者和125例男性健康对照者CYP1A1、GSTM1基因多态性进行检测。结果 CYP1A1 m2位点GG基因型和GSTM1缺失基因型在肺鳞癌组与健康对照者间存在显著性差异(P<0.05 )。吸烟者携带CYP1A1 m2位点至少一个变异G等位基因或携带GSTM1缺失型者患肺鳞癌的危险性进一步显著增加,OR值分别为4.50和3.81(P<0.01)。结论 吸烟与CYP1A1、GSTM1基因多态性与男性肺鳞癌的发生有关。     

HLA-DRB1, -DQA1, -DQB1, -DPA1 and -DPB1 genes in Japanese multiple sclerosis patients

Japanese MS patients and controls were examined for the distribution of HLA-DRB1, -DQA1, -DQB1, -DPA1 and -DPB1 alleles using in vitro amplification of genomic DNA and probing with sequence-specific oligonucleotides. No significant difference in frequency of the examined alleles was observed among the two groups. This is in contrast to Norwegian MS patients, where an association to a combination of certain DQA1 and DQB1 alleles has previously been demonstrated.     

NDE1 and NDEL1: Multimerisation,alternate splicing and DISC1 interaction

Nuclear Distribution Factor E Homolog 1 (NDE1) and NDE-Like 1 (NDEL1) are highly homologous mammalian proteins. However, whereas NDEL1 is well studied, there is remarkably little known about NDE1. We demonstrate the presence of multiple isoforms of both NDE1 and NDEL1 in the brain, showing that NDE1 binds directly to multiple isoforms of Disrupted in Schizophrenia 1 (DISC1), and to itself. We also show that NDE1 can complex with NDEL1. Together these results predict a high degree of complexity of DISC1-mediated regulation of neuronal activity.     

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。     

Rb1cc1 is critical for myoblast differentiation through Rb1 regulation

Rb1-inducible coiled-coil 1 (Rb1cc1) expressed at high levels is associated with the maturation of human embryonic musculoskeletal cells. To clarify the molecular role of Rb1cc1 in muscular differentiation, we investigated the expression of Rb1cc1 and other genes that regulate differentiation in murine embryonic tissues and in C2C12 myoblasts. We also evaluated the effects of RNA interference (RNAi)-mediated Rb1cc1 knockdown on C2C12 myoblast differentiation. After Rb1cc1, Rb1 and myosin heavy chain (Myhc) were expressed in mouse embryonic muscles. The synchronous expression of Rb1cc1 and Rb1 predicted Myhc expression during C2C12 myoblast differentiation. RNAi-mediated knockdown of Rb1cc1 led to Rb1 suppression, and C2C12 myoblasts failed to differentiate. These results indicated that Rb1cc1 is a potent regulator of the Rb1 pathway and a novel mediator that plays a crucial role in muscular differentiation. Rb1cc1 expression is, thus, a prerequisite for myogenic differentiation.     

Maternal genetic polymorphisms in CYP1A1, GSTM1 and GSTT1 and the risk of hypospadias

Hypospadias is one of the most common congenital anomalies. Increased exposure to environmental factors (endocrine-disrupting chemicals and smoking) or maternal endogenous estrogen may cause hypospadias because male sexual differentiation is dependent on normal androgen homeostasis. Moreover, interactions between genetic factors and cigarette smoking and other chemicals have been suggested. It has been demonstrated that the CYP1A1 metabolizes not only environmental chemicals but also estrogens, and glutathione-S-transferases (GSTs) are detoxification enzymes that protect cells from toxicants by conjugation with glutathione. In this study, to investigate the association of CYP1A1 (MspI), GSTM1 and GSTT1 polymorphisms with hypospadias, a case-control study of 31 case mothers who had boys with hypospadias and 64 control mothers was performed in Japan. These polymorphisms were investigated by PCR-based methods using DNA from peripheral lymphocytes. We found that the heterozygous CYP1A1 and heterozygous and homozygous CYP1A1 were less frequent in the case mothers than in the control mothers [adjusted odds ratio (OR)=0.17, 95% confidence interval (CI)=0.04-0.74, OR = 0.28, 95% CI = 0.08-0.97, respectively]. We found no effect of maternal smoking on the hypospadias risks among the gene polymorphisms. The results suggest that mothers with the CYP1A1 MspI variant allele may have a decreased risk for hypospadias.     

Circulatinglong non-coding RNA FEZF1-AS1 and AFAP1-AS1 serve as potential diagnostic biomarkers for gastric cancer

BackgroundGrowing evidence indicates that two long non-coding RNAs (lncRNAs), FEZ family zinc finger 1 antisense RNA 1 (FEZF1-AS1) and Actin filament associated protein 1 antisenseRNA1 (AFAP1-AS1), are highly expressed in different cancers, including gastric cancer (GC). However, the expression pattern and clinical utility of these two lncRNAs are still unknown.MethodsSerum expression levels of FEZF1-AS1 andAFAP1-AS1 were measured by quantitative real-time polymerase chain reaction (qRT-PCR). CEA and CA19-9 were detected by ARCHITET I2000 SR. Analyses were all performed using SPSS software version 20.0 (SPSS Inc., Chicago, USA). P < 0.05 was considered statistically significant.ResultsDetection of serum FEZF1-AS1 and AFAP1-AS1 showed both of them were up-regulated in GC patients compared with the normal controls (p < 0.0001), and high serum expression levels were correlated with tumor size, tumor-node-metastasis (TNM) stage and lymph node metastasis. Besides, the area under the ROC curve (AUC) demonstrated the two lncRNAs had higher diagnostic utility than CEA and CA19-9. Furthermore, when combined the two lncRNAs as a model, it yielded an AUC of 0.866, and the combination of the model, CEA and CA19-9 could observably improve diagnostic sensitivity to 95.5 %. What’s more, circulating FEZF1-AS1 and AFAP1-AS1 were significantly decreased after the GC patients underwent the operation (both p < 0.001).ConclusionOur study indicated that serum FEZF1-AS1 and AFAP1-AS1 had better sensitivity and efficiency for the diagnosis of GC and the combination of the two lncRNAs might be used as a potential prognostic indicator in GC.     

HIV-1 Nef stabilizes AP-1 on membranes without inducing ARF1-independent de novo attachment

HIV-1 Nef affects the trafficking of numerous cellular proteins to optimize viral replication and evade host defenses. The adaptor protein (AP) complexes, which form part of the cytoplasmic coat of endosomal vesicles, are key cellular co-factors for Nef. Nef binds these complexes and alters their physiologic cycle of attachment and release from membranes. Specifically, while AP-1 normally becomes cytosolic when attachment events are blocked by inhibition of the GTPase cycle of ADP-ribosylation factor-1 (ARF1), the complex remains membrane-associated in Nef-expressing cells. To investigate the mechanism of this effect, we used a permeabilized cell system to detect the de novo attachment of exogenous AP-1 to endosomal membranes. Nef did not mediate de novo attachment independently of ARF1, despite its ability to maintain the association of AP-1 with endosomal membranes when the activity of ARF1 was blocked. We conclude that Nef stabilizes AP complexes on endosomal membranes after ARF1-dependent attachment. This stabilization may facilitate coat formation and stimulate the trafficking of multiple cellular proteins.     

神经系统RNA结合蛋白Musashi1 RRM1的初步结晶

目的 对Musashi1发挥功能的 RRM1结构域进行结晶,得到可用来衍射的蛋白晶体,为之后的结构解析打基础。方法 通过构建Musashi1RRM1的原核表达载体,并在BL21中表达、纯化高纯度的蛋白质,通过筛选结晶体条件得到蛋白晶体。结果 通过系统筛选和优化晶体生长条件得到了蛋白晶体。结论 Musashi1 RRM1的蛋白晶体质量较好,满足蛋白晶体衍射和数据收集的要求。     

IL—1受体相关激酶—1和—2协同调控IL—1诱导的AP—1的活化

目的研究白介素 - 1受体相关激酶 - 1(IRAK- 1)和 IRAK- 2在白介素 - 1(IL - 1)诱导 AP- 1活化中的作用。方法L ipofectin介导反义 IRAK- 1寡核苷酸和反义 IRAK- 2寡核苷酸转染 Hep G2细胞。用逆转录 PCR法检测 IRAK - 1和 IRAK- 2m RNA表达水平 ;Western blot分析 IRAK- 1和 IRAK - 2蛋白表达水平。以 Sandwich EL ISA法检测 AP- 1的活化。结果反义IRAK- 1寡核苷酸和反义 IRAK- 2寡核苷酸通过抑制各自靶基因 m RNA和蛋白表达抑制 IL- 1诱导的 AP- 1活化 ;反义 IRAK-1寡核苷酸与反义 IRAK- 2寡核苷酸共转染 Hep G2细胞对 AP- 1的抑制作用较两者单独转染明显增强。结论 IRAK- 1和 I-RAK- 2在调控白介素 - 1诱导的 AP- 1活化时协同作用。     

Ah receptor, CYP1A1, CYP1A2 and CYP1B1 gene polymorphisms are not involved in the risk of recurrent pregnancy loss

The etiology of recurrent pregnancy loss (RPL) remains unclear, but it may be related to a possible genetic predisposition together with involvement of environmental factors. We examined the relation between RPL and polymorphisms in four genes, human aryl hydrocarbon (Ah) receptor, cytochrome P450 (CYP) 1A1, CYP1A2 and CYP1B1, which are involved in the metabolism of a wide range of environmental toxins and carcinogens. All cases and controls were women resident in Sapporo, Japan and the surrounding area. The Ah receptor, CYP1A1, CYP1A2 and CYP1B1 genotypes were assessed in 113 Japanese women with recurrent pregnancy loss (RPL) and 203 ethnically matched women experiencing at least one live birth and no spontaneous abortion (control). No significant differences in Ah receptor, CYP1A1, CYP1A2 and CYP1B1 genotype frequencies were found between the women with RPL and the controls [Ah receptor: Arg/Arg (reference); Arg/Lys and Lys/Lys, odds ratio (OR)=0.67; 95% confidence interval (CI)=0.40-1.11, CYP1A1: m1m1 (reference); m1m2 and m2m2, OR = 0.86; 95% CI = 0.53-1.40, CYP1A2: C/C and C/A (reference); A/A, OR = 1.16; 95% CI = 0.71-1.88, CYP1B1: Leu/Leu (reference); Leu/Val and Val/Val, OR = 1.18; 95% CI = 0.68-2.02]. The present study suggests that the Ah receptor, CYP1A1, CYP1A2 and CYP1B1 gene polymorphisms are not major genetic regulators in RPL.