天津地区汉族人2型糖尿病与维生素D受体基因多态性的研究

目的:探讨天津地区汉族人2型糖尿病(T2DM)与维生素D受体(VDR)基因多态性的关系。方法:采用聚合酶链反应-限制性酶切片段长度多态性分析方法测定116例T2DM患者和112例正常对照者的VDR-Fok1和VDR-Bsm1位点基因型,计算并比较T2DM患者和正常对照组基因型频率。结果:T2DM患者与正常对照组VDR-Fok1位点基因型、F/f等位基因的分布差异无统计学意义(P>0.05);VDR-Bsm1位点基因型、B/b等位基因的分布差异有统计学意义,T2DM患者VDR-Bsm1位点Bb基因型、B等位基因明显增加(P<0.01)。结论:VDR基因Bsm1位点Bb基因型、B等位基因可能是天津地区汉族人T2DM的基因型和易感基因。     

对氧磷酯酶1与2型糖尿病心脑血管并发症的关系

目的 探讨血清对氧磷酯酶1(PON1)活性与2型糖尿病(DM)心脑血管并发症的关系.方法 对2型糖尿病心脑血管并发症患者和正常对照组以对氧磷为底物测定血清PON1活性.结果 单纯2型糖尿病组PON1活性为(152±67)IU/L,比正常对照组显著降低[(224±89)IU/L,P＜0.01],2型糖尿病并发心脑血管病变组PON1活性为(110±46)IU/L,比单纯2型糖尿病组显著降低(P＜0.01).结论 PON1活性在单纯2型糖尿病及并发心脑血管病变患者中均显著降低,提示PON1活性的降低参与了糖尿病心脑血管并发症的发生.     

中国汉族人群对氧磷酶Q192R基因多态性与冠心病的相关性及危险因素对基因型影响的研究

目的:测定对氧磷酶(PON1)Q192R基因多态性及在人群中的分布,并分析不同基因型与冠心病(CHD)的关系,CHD常见危险因素对基因型亚组的影响。方法:应用聚合酶链反应-限制性片断长度多态性(PCR-RFLP)分析机聚丙稀酰胺凝胶电泳方法检测PON1多态性,等位基因频率计算采用计数法。对所得结果采用t检验、t′检验、或χ2检验进行统计学处理,统计分析用Sars软件。结果:PON1Q192R有QQ、QR、RR三种基因型,正常人的基因型频率分别为0.2000、0.4909、0.3091,Q、R等位基因频率分别为0.4500、0.5500,冠心病组QQ、QR、RR三种基因频率分别为0.2266、0.5313、0.2421,Q、R等位基因频率分别为0.4921、0.5079。吸烟对含R等位基因的基因型QR、RR是冠心病的独立危险因素,在冠心病组和对照组中血糖、血脂、饮酒、血压在各亚组中的影响均无明显相关性。结论:中国汉族人群(潍坊地区)PON1基因多态性和冠心病无关。吸烟可能是含R基因的基因型的人群患冠心病的易感因素。     

内皮固有型一氧化氮合酶基因4b/a多态性与2型糖尿病并发冠心病的关联性研究

目的 本研究探讨内皮固有型一氧化氮合酶(ecNOS)基因第4内含子27bp可变数目串联重复序列(VNTR)插入/缺失多态性(4b/a)与2型糖尿病(T2DM)并发冠心病的关联性.方法 观察80例2型糖尿病、92例T2DM合并冠心病患者及119例正常对照组人群,提取基因组DNA并应用PCR技术检测ecNOS基因内含子4VNTR多态性.结果 ecNOS基因4b/a多态性与T2DM及T2DM并发冠心病无关.发现1例4a/a纯合子基因型者,位于T2DM并发冠心病组.结论 缺失型等位基因4a可能处于非统治地位,是否与T2DM并发冠心病有关仍待更大样本的观察.     

内皮型一氧化氮合酶基因多态性与2型糖尿病合并高尿酸血症相关性研究

目的:探讨汉族人内皮型一氧化氮合酶(eNOS)基因内含子4可变数目串联重复序列插人,缺失多态性(4b/a)与2型糖尿病(T2DM)合并高尿酸血症的关系.方法:提取100例健康人(A组)、76例T2DM无高尿酸血症患者(B组)和124例T2DM合并高尿酸血症患者(C组)的基因组DNA,用聚合酶链反应-可变数目串联重复序列法测定4b/a多态性.结果:3组eNOS基因内含子4等位基因的分布和频率差异有统计学意义(P<0.05).T2DM所有患者中4b/b、4b/4a、4a/4a各基因型高尿酸血症的发病率逐渐升高分别为35.4%、58.8%和100%.4b和4a等位基因患者高尿酸血症的发病率分别为36.5%和63.6%,两者比较差异有统计学意义.携带a等位基因的T2DM患者较未携带a等位基因者合并高尿酸血症者的危险性升高(4b/4a的OR为3.42,P<0.05).结论:eNOS 4a等位基因是T2DM患者并发高尿酸血症的独立危险因素,可能为筛查T2DM患者高尿酸血症的高危人群提供侯选基因.     

贵州地区汉族人群2型糖尿病肾病RANTES基因启动子多态性分析

目的探讨RANTES基因启动子-28C/G多态性与贵州地区汉族人群2型糖尿病并发肾病之间的关系。方法应用聚合酶链反应限制性片段长度多态性技术(PCR-RFLP),检测143例2型糖尿病患者(53例单纯2型糖尿病患者,90例2型糖尿病合并肾病患者)和55例正常对照组RANTES基因启动子-28C/G基因型,并对各组间的等位基因频率与基因型频率进行检测分析。结果(1)正常对照组与2型糖尿病组相比,-28G阳性基因型频率和G等位基因型频率差异无显著意义(P>0.05)。(2)糖尿病肾病组与单纯糖尿病组相比,-28G阳性基因型频率和G等位基因型频率显著升高(P<0.05)。结论RANTES基因启动子-28G等位基因与贵州地区汉族人群2型糖尿病的发生无关,但可能是糖尿病肾病的易感基因。     

MMP-9-1562C/T基因多态性与2型糖尿病肾病关系的研究

目的:探讨2型糖尿病(T2DM)患者基质金属蛋白酶-9基因-1562C/T(MMP-9-562C/T)多态性与糖尿病肾病(DN)及DN不同病期的关系.方法:将150例T2DM患者按DN诊断标准分为 DN 组与非糖尿病肾病(NDN)组,DN组分为微量白蛋白尿期、临床白蛋白尿期和肾功能不全期.另择52例健康人作为正常对照(NC)组.应用限制性片段长度多态性(RFLP)分析各组基因型.结果:DN组的糖化血红蛋白(HbA1c)、收缩压(SBP)、肌酐(Cr)、尿素氮(BUN)、尿微量白蛋白排泄率(UAER)值高于 NDN、NC组(P<0.01).DN组与NDN 组、NC组之间基因型分布差别均有统计学意义(P<0.01).T2DM中CC、CT、TT各基因型DN发生率依次递减,其中TT基因型组DN发生率低于CC基因型组(P<0.01).携 T 等位基因者发生DN的风险是携C等位基因者的0.47倍 (P<0.01,95%CI 0.29～0.75).MMP-9-1562 C基因型、HbA1c、TG、TC、Cr、BUN、SBP是DN发生的危险因素;DN组内微量白蛋白尿期、临床自蛋白尿期和肾功能不全期TT基因型与T等位基因频率随肾功能减退而降低(P<0.05或P<0.01).结论:MMP-9-1562C/T 基因多态性与DN发生有关,T等位基因是DN患者的保护基因.     

趋化因子CXCL5启动子区-156G/C基因多态性与2型糖尿病的关系

目的:探讨趋化因子CXCL5基因启动子-156位点G/C单核苷酸多态性与2型糖尿病的关系。方法:随机选择我院2型糖尿病患者(T2DM组)210例,对照组171例。采用聚合酶链式反应限制性片段长度多态性(PCR-RFLP)方法,检测趋化因子CXCL5基因启动子-156位点G/C多态性。结果:趋化因子CXCL5启动子-156G/C多态性(各基因型频率、C等位基因频率)在T2DM组与对照组的分布差异无统计学意义(P>0.05)。结论:CXCL5基因多态性可能不是糖尿病发病的危险因素。     

动脉粥样硬化性脑梗死合并2型糖尿病对氧磷酶2基因多态性相关分析

目的探讨对氧磷酶（PON）2基因多态性与动脉粥样硬化性脑梗死、2型糖尿病的相关性。方法将80例动脉粥样硬化性脑梗死患者按是否合并糖尿病分为脑梗死组（55例）和脑梗死合并糖尿病组（25例）2个亚组,同期选择门诊体检的健康体检者80名为健康对照组。2型糖尿病患者明确诊断、采用聚合酶链反应-限制性内切酶片段长度多态性（PCR-RFLP）分析PON基因中PON2A148G、C311S基因多态性,进行差异性分析。结果缺血性脑梗死合并2型糖尿病（T2DM）组与缺血性脑梗死组及健康对照组相比,PON2A148G基因型分布差异无统计学意义、PON2C311S基因型分布差异有统计学意义。结论PCIN2C311S多态性C等位基因的存在可能与缺血性脑卒中患者合并2型糖尿病的概率增高相关。     

2型糖尿病8-异前列腺素F2α与脂联素基因SNP+45（T/G）多态性的相关性

目的:探讨脂联素基因SNP+45(T/G)多态性与2型糖尿病(T2DM)氧化应激指标8-异前列腺素F2α之间的关系。方法:聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)方法检测120例T2DM患者(T2DM组)脂联素SNP+45(T/G)多态性,同时选取正常体检者60例为对照组,采用竞争酶免疫测定的方法检测T2DM患者血清8-异前列腺素F2α水平,检测各组空腹血糖、血脂及空腹胰岛素水平,计算HOMA-IR等指标。结果:T2DM患者血清8-异前列腺素F2α水平高于对照组,T2DM患者脂联素SNP+45位点GG基因型频率高于对照组(P<0.05)。T2DM组内GG型的体质量指数(BMI)高于TT及TG型(P<0.05);TT、TG及GG基因型血清8-异前列腺素F2α水平差异无统计学意义(P>0.05)。结论:脂联素SNP+45(T/G)多态性与T2DM氧化应激之间可能无相关关系。     

Vascular sphingosine-1-phosphate S1P1 and S1P3 receptors

The sphingolipid sphingosine-1-phosphate (S1P) acts on five subtypes of G-protein- coupled receptors, termed S1P(1) (formerly endothelial differentiation gene-1 [Edg-1]), S1P(2) (Edg-5), S1P(3) (Edg-3), S1P(4) (Edg-6) and S1P(5) (Edg-8), and possibly several other "orphan" receptors, such as GPR3, GPR6 and GPR12. These receptors are coupled to different intracellular second messenger systems, including adenylate cyclase, phospholipase C, phosphatidylinositol 3-kinase/protein kinase Akt, mitogen-activated protein kinases, as well as Rho- and Ras-dependent pathways. Consistently with this receptor multiplicity and pleiotropic signaling mechanisms, S1P influences numerous cell functions. S1P(1)1, S1P(2) and S1P(3) receptors are the major S1P receptor subtypes in the cardiovascular system, where they mediate the effects of S1P released from platelets, and possibly other tissues (such as brain). Thus S1P(1) and S1P(3) receptors enhance endothelial and vascular smooth muscle cell proliferation and migration, playing a key role in developmental and pathological angiogenesis. In contrast, S1P(2) receptors inhibit migration of these cell types, probably because of their unique stimulatory effect on a GTPase-activating protein inhibiting the activity of Rac. S1P receptors can also cause relaxation and constriction of blood vessels. The former effect is mediated by pertussis toxin-sensitive receptors (possibly S1P(1)) located on the endothelium and stimulating phosphatidylinositol 3-kinase/Akt/endothelial nitric oxide synthase (eNOS). The vasoconstricting effect of S1P is likely to be mediated by S1P(2) and/or S1P(3) receptors, via Rho-Rho-kinase, and is more potent in coronary and cerebral blood vessels. Finally, S1P also protects endothelial cells from apoptosis through activation of phosphatidylinositol 3-kinase/Akt/eNOS via S1P(1) and S1P(3) receptors. The variety of these effects, taken together with the existence of multiple receptor subtypes, provides an abundance of therapeutic targets that currently still await the development of selective agents.     

Antimykobakterielle 1-Phenyl-1-alkylaminoalkane

Antimycobacterial 1-Phenyl-1-alkylaminoalkanes Synthesis and testing for antimycobacterial properties (M. tuberculosis H 37 Ra, Middlebrook-7H9-broth) of 1-phenyl-1-alkylaminoalkanes, which differ from antimycobacterial N-alkylbenzylamines by an additional alkyl chain in α-position, is described. By variation of both alkyl chains and introduction of one or two Cl-substituents in the aromatic ring the activity increases up to an optimum within the homologous series. Overstepping optimal lipophilicity or ramification of the alkyl chains decrease activity. Compounds 19, 20, 33-35, 51-53, 61-63, 65-67, 70-73, 96 and 102 - 104 inhibit the growth of M. tuberculosis in concentrations of 2 to 4 μg/ml.     

Sulfation of iodothyronines by human sulfotransferase 1C1 (SULT1C1)*

Sulfation is an important component of human thyroid hormone metabolism. The role of the human sulfotransferase 1C1 (SULT1C1) is not known. Because SULT1C1 is present in the adult thyroid, intra-thyroidal sulfation of thyroid hormones and their metabolites might occur. We tested this hypothesis by determining the ability of recombinant human SULT1C1 to catalyze iodothyronine sulfation. Apparent K(m) values for 3,3',5-triiodothyronine (T(3)), 3, 3'-diiodothyronine (3,3'-T(2)), 3',5',3-triiodothyronine (rT(3)), and 3,3',5,5'-tetraiodothyronine (T(4)) with SULT1C1 were 28.7, 10.3, 10.2, and 59.3 microM, respectively. Thermal stability and responses to inhibitors also were tested with T(3) as the substrate. Enzyme aliquots were measured simultaneously to determine SULT1C1 substrate preferences at optimal iodothyronine concentrations. SULT1C1 activity obtained with T(3) was used as 100%, and the activities with 3,3'-T(2), rT(3), T(4), and 3,5-diiodothyronine (3, 5-T(2)) were 614, 314, 25, and 4%, respectively. We report for the first time the characterization of human SULT1C1 with T(3) and the preferences of the enzyme for various iodothyronines. The presence of SULT1C1 in the adult thyroid gland raises the possibilities that the enzyme can contribute to intraglandular thyroid hormone processing and iodide reutilization.     

Phenotype of the Cyp1a1/1a2/1b1-/- triple-knockout mouse

Crossing the Cyp1a1/1a2(-/-) double-knockout mouse with the Cyp1b1(-/-) single-knockout mouse, we generated the Cyp1a1/1a2/1b1(-/-) triple-knockout mouse. In this triple-knockout mouse, statistically significant phenotypes (with incomplete penetrance) included slower weight gain and greater risk of embryolethality before gestational day 11, hydrocephalus, hermaphroditism, and cystic ovaries. Oral benzo[a]pyrene (BaP) daily for 18 days in the Cyp1a1/1a2(-/-) produced the same degree of marked immunosuppression as seen in the Cyp1a1(-/-) mouse; we believe this reflects the absence of intestinal CYP1A1. Oral BaP-treated Cyp1a1/1a2/1b1(-/-) mice showed the same "rescued" response as that seen in the Cyp1a1/1b1(-/-) mouse; we believe this reflects the absence of CYP1B1 in immune tissues. Urinary metabolite profiles were dramatically different between untreated triple-knockout and wild-type; principal components analysis showed that the shifts in urinary metabolite patterns in oral BaP-treated triple-knockout and wild-type mice were also strikingly different. Liver microarray cDNA differential expression (comparing triple-knockout with wild-type) revealed at least 89 genes up- and 62 genes down-regulated (P-value < or = 0.00086). Gene Ontology "classes of genes" most perturbed in the untreated triple-knockout (compared with wild-type) include lipid, steroid, and cholesterol biosynthesis and metabolism; nucleosome and chromatin assembly; carboxylic and organic acid metabolism; metal-ion binding; and ion homeostasis. In the triple-knockout compared with the wild-type mice, response to zymosan-induced peritonitis was strikingly exaggerated, which may well reflect down-regulation of Socs2 expression. If a single common molecular pathway is responsible for all of these phenotypes, we suggest that functional effects of the loss of all three Cyp1 genes could be explained by perturbations in CYP1-mediated eicosanoid production, catabolism and activities.     

间质作用因子1通过正调控窖蛋白1抑制FBJ-S1-H的迁移性

目的证明间质作用因子(stromal interaction molecule1,Stim1)在FBJ诱导的小鼠骨肉瘤细胞中的抑癌作用。方法在Stim1高表达的FBJ-S1-H细胞采用Stim1以siRNA干扰技术得到Stim1沉默的几株S1-H单克隆细胞株,通过细胞行为学方法和RT-PCR技术对其mRNA进行研究,通过明胶酶谱法对细胞基质金属酶活性进行研究。结果通过细胞行为学方法证明,Stim1的沉默提高了细胞的迁移性,通过对mRNA表达的研究发现,Stim1沉默引起了多种基因表达的变化,其中包括基质金属酶9(matrix mexalloprotelnase 9,MMP-9)的升高,窖蛋白(caveolinl,Cav1),甾醇调控因子Srebf1的降低等,提高单克隆细胞中的Cav1含量可以使细胞迁移性降低。结论实验结果证明在FBJ-S1-H细胞中,Stim1能够抑制细胞的移动性,沉默Stim1的表达能够提高细胞的迁移性。