血管紧张素Ⅰ转化酶及血管紧张素Ⅱ受体1型基因多态性与糖尿病肾病的关联研究

目的探讨血管紧张素I转化酶(angiotensin converting enzyme,ACE)基因I/D多态性及血管紧张素Ⅱ受体1型(angiotensinⅡtype 1 receptor,AT1R)基因A1166C多态性与糖尿病肾病的关系。方法选择2013年4月至2015年10月在广东医科大学附属医院、东莞市塘厦医院和东莞市大朗医院住院2型糖尿病患者,研究对象分为2型糖尿病组(T2DM组)97例和正常对照组50例,其中T2DM组根据有无肾病分为52例糖尿病肾病(DN)组和45例糖尿病非肾病(非DN)组;采用标准的酚-氯仿-蛋白酶K法提取外周血DNA;分别用聚合酶链反应(PCR)技术、PCR-限制性片段长度多态性技术(PCR-RFLP)对ACE I/D、ATlR Al166C多态性进行检测。遗传平衡吻合度检验用HardyWeinberg平衡法。采用χ~2检验分析基因多态性与DN的相关性。结果 DN组和非DN组基线资料无统计学差异(P0.05)。经χ~2检验,DN组DD基因型频率32.6%(17/52)明显高于非DN组DD基因型频率20.0%(9/45)(χ~2=6.62,P0.05);DN组D等位基因频率58.7%(61/104)明显高于非DN组D基因型频率41.1%(37/90)(χ~2=5.94,P0.05)。DN组与非DN组AT1R Al166C各基因型分布频率无显著性差异(χ~2=0.22,P0.05);各等位基因频率分布亦无明显差异(χ~2=0.21,P0.05)。对照组与T2DM组ACE I/D和ATlR Al166C基因多态的基因型分布和等位基因频率均无显著性差异(P0.05)。结论 ACE基因I/D多态性与DN有关,携带DD基因型和D等位基因的T2DM患者是DN的易感人群。AT1R基因A1166C多态性与T2DM并发DN无关。     

内皮细胞型一氧化氮合酶基因多态性与前列腺癌的相关性研究

目的 探讨内皮细胞型一氧化氮合酶(ecNOS)基因多态性与前列腺癌(PCa)是否存在相关性.方法 分别采用聚合酶链反应-限制性片段长度多态性方法(PCR-RFLP)和序列特异性引物扩增方法(PCR-SSP),检测78例PCa患者和88例前列腺增生(BPH)患者的ecNOS基因G894T和ecNOS 4a/b VNTR多态性,分析两组ecNOS G894T和ecNOS 4a/b VNTR的基因型频率与等位基因频率的差异.结果 ecNOSG894T存在G、T 2种等位基因以及GG、GT、TT 3种基因型,ecNOS4a/b VNTR存在a、b 2种等位基因以及aa、ab及bb 3种基因型.PCa组与BPH组相比,ecNOS G894T和ecNOS4a/b VNTR的基因型频率及等位基因频率差异无统计学意义(P均>0.05).结论 ecNOS基因多态性不是前列腺癌发病的遗传危险因素.     

MTHFR基因多态性与2型糖尿病肾病相关性的研究

目的 研究亚甲基四氢叶酸还原酶（MTHFR）基因多态性与2型糖尿病肾病（DN）易感性的关系.方法 选择111例山西地区汉族人2型糖尿病患者,其中糖尿病肾病（DN＋）组56例,糖尿病非肾病（DN-）组55例,运用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）技术并结合琼脂糖凝胶电泳的方法检测111例患者的MTHFR基因多态性,测定各组间基因型频率和等位基因频率.结果 纯合基因型TT、T等位基因在DN＋组（21.43％,46.43％）的频率均明显高于DN-组（7.27％,29.09％）,差异均具有统计学意义（P＜0.05）.无论是在DN＋组还是DN-组中,TT基因型患者血同型半胱氨酸（Hcy）平均水平均大于CC基因型和CT基因型患者,DN＋组血浆Hcy水平明显高于DN-组,差异均具有统计学意义（P＜0.05）.在叶酸浓度≤6.92nmol/L时,DN＋组（24.24％,48.49％） TT型发生率及T等位基因频率明显高于DN-组（3.70％,25.93％）（P＜0.05）,当叶酸浓度＞6,92nmol/L时,DN＋组TT型发生率及T等位基因频率与DN-组无差异（P＞0.05）.结论 MTHFR基因C677T多态性与糖尿病肾病（DN）发生具有相关性,突变的T等位基因是DN易感基因,但其影响效果受叶酸浓度的影响.     

醛糖还原酶基因5＇端(A-C)n的多态性与Ⅱ型糖尿病患者糖尿病肾病易感性的关系

目的探讨Ⅱ型糖尿病（DM）醛糖还原酶（AR）基因5’端的（AC）n的多态性与糖尿病肾病（DN）易感性的关系。方法应用32PdCTP掺入聚合酶链反应（PCR）,PCR产物6％甲酰胺聚丙烯酰胺凝胶电泳及放射性自显影的方法,分析了113例Ⅱ型DM组和42例正常对照（CON）组醛糖还原酶（AR）基因5’端的（AC）n的多态性与糖尿病肾病易感性的关系。结果DM组和对照组均发现7种等位基因（Z－6～Z＋6）,Z－2等位基因频率在DN组高于无微血管并发症组（NDC）和CON组;Z＋2等位基因频率在DN组低于NDC和CON组,P＜0．05。结论中国北方汉族Ⅱ型DM患者AR基因5’端（AC）n存在7种等位基因（Z－6～Z＋6）。Z－2等位基因可能是DN的易感基因,Z＋2等位基因可能为其保护基因。     

2型糖尿病肾病患者p22phox基因多态性的研究

目的研究NADPH氧化酶p22phox亚基基因多态性与中国上海汉族人群2型糖尿病肾病（DN）相关性。方法应用限制性片断长度多态性（RFLP）-PCR方法对105例健康对照组和194例2型糖尿病（DM）患者（其中71例DN）进行p22phox亚基C242T、A640G基因型检测。同时检测其身高、体重、血压、血脂、空腹血糖及胰岛素、HbAlc的水平。结果DN组CT＋TT基因型频率明显高于2型DM和对照组（26．76％比17．07％、3．81％，P=0．0002）；DN组T等位基因频率明显高于2型DM和对照组（22．54％比13．42％、2．86％，P=0．0001）；3组间AA基因型频率与A等位基因频率差异无统计学意义。多元回归分析显示，T242等位基因、收缩压、空腹血糖、HbAlc、β细胞功能指数（Homa-IS）是DN的危险因素。结论p22phox亚基T242等位基因变异可能是中国上海地区汉族人群DN的易感基因：而p22phox亚基A640G基因多态性与上述人群DN无相关性。T242等位基因、收缩压、空腹血糖、HbAlc、Homa-IS是DN的危险因素。     

维生素D受体基因BsmI多态性与2型糖尿病肾病的关系

目的 研究维生素D受体(VDR)基因BsmI位点多态性与汉族人群2型糖尿病肾病(DN)的关系.方法 应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测304例2型糖尿病患者(DM组)及100例健康体检者(NC组)VDR Bsml位点基因型和等位基因频率.根据尿白蛋白情况将DM组分为非糖尿病肾病组(DN0组,122例)、微量白蛋白尿组(DN1组,87例)、大量白蛋白尿组(DN2组,95例).83例病程5年以上仍未出现肾病的DM患者纳入L-NDN组;64例起病1年内即出现肾病的DM患者纳入EDN组.结果 DM组BB+Bb基因型和B等位基因频率均高于NC组(x2=7.088,P=0.008;x2=5.865,P=0.015).DN2组BB+Bb基因型和B等位基因频率高于NC组(x2=14.287,P=0.000; x2=12.621,P=0.000)及DN0组(x2=8.063,P=0.005;x2=8.173,P=0.004).其余组间差异均无统计学意义.EDN组BB+Bb基因型和B等位基因频率均显著高于L-NDN组(x2=7.228,P=0.007;x2=5.853,P=0.016).B等位基因阳性DN患者的尿白蛋白排泄率显著高于B等位基因阴性DN患者,差异有统计学意义(P＜0.01).BsmI位点基因型与DN发生密切相关.B等位基因阳性是DN发生及早发的危险因素(OR=2.004;0R=2.394).结论 VDRBsmI基因多态性与DN易感性相关.B等位基因阳性患者更易出现大量白蛋白尿及早期发生肾病.     

Ⅱ型糖尿病肾病患者β3—肾上腺素能受体基因多态性研究

目的：探讨β3-肾上腺素能受体（β2-AR）基因64位点的色氨酸（Trp)密码子被精氨酸（Arg)置换与糖尿病肾病（DN）之间的关系。方法;采用聚合酶链反应-限制性片段长度多态性（PCR-RELP）技术,研究了196例Ⅱ型糖尿病患者β3-AR基因Trp64Arg突变基因型,并测体重指数（BMI）、血压、尿微量白蛋白排泄率（UAER）、空腹血脂、血糖、糖化血红蛋白A1c(HbA1c)、腹及餐后2hC肽。结果：⑴中国上海地区Ⅱ型糖尿病患者Arg64等位基因频率为0.13,低于印第安人（P＜0.001)和日本人（P＜0.01),略高于芬兰人（P＞0.05)。⑵糖尿病患者中,Arg64携带者组BMI、血甘油三酯水平明显高于非携带者组（P＜0.05),而发病年龄、确诊糖尿病时年龄、病程、血压、空腹血糖、HbA1c、空腹及餐后2hC肽、总胆固醇、高密度脂蛋白及低密度脂蛋白水平,两组无明显差别。⑶DN组Arg64等位基因频率为0.18,明显高于非DN组（P＜0.01),且与DN分期无关。结论：中国上海地区Ⅱ型糖尿病患者β3-AR基因Trp64Arg突变与体内脂肪分布及脂代谢紊乱有关;Arg64等位基因可能是DN的一个风险基因。     

内皮型一氧化氮合酶Glu298Asp基因多态性与老年原发性高血压微量白蛋白尿的关系

目的研究内皮型一氧化氮合酶（eNOS）Glu298Asp基因多态性与老年原发性高血压（EH）微量白蛋白尿（MAU）的关系。方法从到医院就诊的老年EH患者中筛选出202例无显性蛋白尿的患者,行24h MAU测定,并应用基因芯片技术检测eNOS Glu298Asp基因多态性,按照24hMAU定量分为MAU组和非MAU组（NAU组）,比较两组基因型和等位基因分布差异。结果两组等位基因和基因型的分布不同,MAU组等位基因T及含等位基因T的基因型（GT＋TT）分布频率明显升高（χ^2=6.62,P〈0.01;χ^2=7.29,P〈0.01）;T等位基因变异使老年EH患者MAU的相对危险度显著增高（OR=2.361,95%CI=1.256～4.437）。结论 T等位基因是老年EH患者MAU的易感基因,携带T等位基因导致老年EH患者出现MAU的风险显著增高。     

甲状旁腺素基因多态与2型糖尿病患者骨密度的关系

目的 探讨甲状旁腺素(PTH)基因多态与中国北方汉族人2型糖尿病患者骨密度的关系.方法 运用聚合酶链反应限制性片段长度多态性(PCR-RFLP)技术检测了2型糖尿病(T2DM)组104例,健康对照(CON)组102例,206例中国北方汉族人PTH基因多态性;采用双能X线吸收法骨密度仪(DEXA)测量骨密度(BMD).结果 甲状旁腺素(BSTB1位点)基因型和等位基因分布频率在2型糖尿病组与对照组间差异无显著性(P>0.05);在对照组及2型糖尿病组,Bb/bb基因型者L2-4和Neck部位BMD显著低于BB型(P<0.05),Troch及Wards三角区BMD差异无显著性(P>0.05).结论 2型糖尿病患者PTH基因多态性(BST B1位点)可能是预测骨量减少、骨质疏松易感性的遗传标志.     

细胞间黏附分子1基因K469E多态性与2型糖尿病肾病的相关性研究

目的 探讨细胞间黏附分子1(ICAM-1) K469E基因多态性与2型糖尿病肾病(DN)的关系。方法 运用PCR-限制性多态性片断长度(RFLP)技术检测98名健康对照者和210例2型糖尿病患者(其中DN患者110例，DM患者100例)的基因型，比较各组的基因型和等位基因频率。结果 (1)DN组KK基因型和K等位基因频率显著高于DM组和正常对照组；(2)与EE型组相比，KK型组DN的发生率明显上升；(3)Logistic回归分析显示ICAM-1 K469E基因多态性是DN的危险因素。结论 ICAM-1 K469E多态性与2型糖尿病伴发肾病的发生有关，K等位基因可能是DN的易感基因。     

对氧磷酯酶基因多态性与糖尿病肾病的关系

目的探讨对氧磷酯酶2(PON2)基因A148G多态性与糖尿病肾病的关系.方法(1)用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析法探查PON2基因A148G多态性在正常对照组、单纯2型糖尿病组、糖尿病肾病组中的基因频率分布;(2)放射免疫法检测血清免疫反应性胰岛素(IRI)、C肽(C-P)水平.结果(1)糖尿病肾病组GG基因型和G等位基因频率明显高于单纯2型糖尿病组(X2=4.26 P＜0.05,X2=4.89P＜0.05)和正常对照组(X2=4.79 P＜0.05,X2=5.49P＜0.01);(2)基因型为GG的糖尿病患者空腹血糖浓度高于基因型为GA和AA的糖尿病患者的空腹血糖浓度(F=3.90 P＜0.05,F=4.23 P＜0.05);(3)Logistic回归分析表明GG基因型是糖尿病肾病的独立变异危险因素(P＜0.05).结论PON2基因多态性与糖尿病肾病的发生有关.     

Association of the nitric oxide synthase gene polymorphism with an increased risk for progression to diabetic nephropathy in type 2 diabetes

A mutation of endothelial nitric oxide synthase (ecNOS)-a key enzyme of the endogenous nitrovasodilator system that is essential for the regulation of blood flow and blood pressure-may aggravate the progression to diabetic nephropathy and/or retinopathy. To investigate the association of ecNOS tandem repeat polymorphism with diabetic nephropathy, the ecNOS genotype was assessed in 82 Japanese type 2 diabetic patients without nephropathy, 94 patients with microalbuminuria, 39 patients with nephropathy, and 155 healthy control subjects. The analysis revealed that type 2 diabetic patients with nephropathy (not with microalbuminuria) were significantly different from type 2 diabetic patients without nephropathy and healthy control subjects in genotype distribution (P = 0.0423) and frequency of the ecNOS4a allele (19.2% vs. 7.3 and 7.1%, respectively; P = 0.0078). The odds ratio of progression to diabetic nephropathy in diabetic patients who carry the mutated allele is about 2.87 compared with noncarriers. The stepwise multiple regression analysis in these patients showed that hypertension (F = 9.760) and ecNOS gene polymorphism (F = 5.298) are the relevant variables for nephropathy. However, no association was found between the ecNOS4a allele and hypertension or diabetic retinopathy. These results imply that the ecNOS gene polymorphism may be associated with progression to diabetic nephropathy in Japanese type 2 diabetic patients.     

西藏地区糖尿病肾病临床特点分析

目的 分析西藏地区2型糖尿病肾病(DN)的临床特点。 方法 回顾分析2001年5月至2006年10月间在我科住院的306例2型糖尿病（DM）患者的临床资料。 结果 306例DM患者包括151例DN和155例非DN患者，根据尿白蛋白及Scr水平，DN组患者再分为微量白蛋白尿组、临床蛋白尿组和肾功能不全组。DN组尿微量白蛋白、Scr和血、尿β2微球蛋白(MG)均较非DN组显著增高（均P < 0.01）；且尿微量白蛋白与收缩压、血β2-MG呈正相关（r = 0.187， P < 0.05； r = 0.297， P < 0.01），而与GFR呈负相关（r = －0.287，P < 0.01）。DN组高血压发生率高（60.27％），血压显著高于非DN组（P < 0.01），且以收缩压更显著。DN组发生尿毒症者14例（9.27％），死亡8例（5.30％），其中5例死于尿毒症；并发糖尿病视网膜病变20例(13.25%)；发生心脑血管意外者6例(3.97%)。 结论 西藏地区2型糖尿病肾病早期即有明显的蛋白尿、血压及血、尿β2-MG增高，后期GFR急剧下降且并发症多而严重。     

Correlation between parathyroid hormone, bone alkaline phosphatase and N-telopeptide of type 1 collagen in diabetic and non-diabetic haemodialysis patients

BACKGROUND: Haemodialysis (HD) patients with diabetes mellitus often have renal osteodystrophy (ROD) characterized by reduced bone turnover, but little is known about the correlation between bone formation and bone resorption in this population. METHODS: The authors measured serum parathyroid hormone (PTH), bone alkaline phosphatase (BAP), N-telopeptides of type 1 collagen (NTx) and fasting glucose in 48 patients with diabetic nephropathy (DN) and 80 patients with glomerulonephritis (non-DN) who had received or=5 years HD (r = 0.568) this correlation was similar to that in the non-DN group (r = 0.653), whereas there was no significant correlation in those receiving <5 years HD. Patients receiving >or=5 years HD had a comparable glucose level (111.1 +/- 19.2 mg/dL) to the non-DN group, whereas those receiving <5 years had a higher level (196.1 +/- 53.1 mg/dL). CONCLUSION: Differences in the interaction between bone cells between DN and non-DN patients are one potential cause of lower bone turnover in the former group. Research of this correlation is needed to increase understanding of the complexities of bone metabolism in DN patients.     

Haplotype analysis of the RAGE gene: identification of a haplotype marker for diabetic nephropathy in type 2 diabetes mellitus.

BACKGROUND: Diabetic nephropathy (DN) represents a devastating complication of diabetes. Family clustering, heterogeneity in the onset and progression and results of segregation studies indicate that susceptibility to DN is a complex trait. METHODS: Common single nucleotide polymorphisms in the RAGE (receptor of advanced glycation end-products) gene (-429T/C, -374T/A, G82S, 1704G/T, 2184A/G and 2245G/A) were studied in the association study comprising 605 Caucasian subjects by means of haplotype analysis in order to identify an eventual haplotype marker for DN in type 2 diabetes. Haplotypes were constructed computationally; frequencies were compared among groups of subjects with type 2 diabetes (DM) and DN, diabetics without DN and non-diabetics. Survival analysis was carried out to ascertain whether certain RAGE haplotypes influence onset of DN in type 2 diabetics. RESULTS: Significant differences in haplotype frequencies among DM + DN vs DM non-DN and non-DM groups were found (P = 0.0007 and 0.0013, respectively; permutation test). Frequency of the RAGE(2) haplotype containing minor alleles in positions -429 and 2184 (CTGGGG) in the DN group was significantly higher than in the two control groups (21.7% vs 12.8% and 13.8%, both P(corr)<0.003; two-tail Fisher exact test); odds ratios 1.65 [95% confidence interval (CI): 1.08-2.50; P = 0.020] and 1.79 (95% CI: 1.22-2.62; P = 0.003), respectively. In survival analysis, duration of diabetes until the onset of DN (e.g. appearance of persistent proteinuria) was significantly different among RAGE(2) diplotype groups (P<0.05); median DN-free interval was 9.6 years in RAGE(2) +/+ homozygotes, 15.2 years in +/- heterozygotes and 17.0 years in the -/- combination. CONCLUSIONS: The RAGE(2) haplotype is associated with DN in type 2 diabetics and with earlier DN onset and, thus, can be regarded a marker for DN.     

A polymorphism in the gene for the atrial natriuretic peptide and diabetic nephropathy

Background: Atrial natriuretic peptide is involved in blood pressure regulation via its vasodilating and natriuretic actions. Since diabetic nephropathy and hypertension are closely related, ANP is a reasonable candidate gene for diabetic nephropathy (DN). Methods: We genotyped 410 patients with type I diabetes (without DN n=307; with DN n=103) and 658 patients with type II diabetes (without DN n=464; with DN n=194). In the patients the duration of diabetes was at least 10 years. Diabetic nephropathy was defined as urinary albumin excretion of ⩾30 mg/24 h. The Hpa II polymorphism in intron 2 of the ANP gene was determined using PCR amplification followed by restriction digest. Alleles were separated on agarose gels stained with ethidium bromide. Results: We compared genotype distribution and allele frequencies between patients with and without nephropathy. No significant difference was observed either in type I (allele frequency without DN H1, 0.02/H2, 0.98 vs with DN H1, 0.05/H2, 0.95; P=0.132) or in type II diabetes (allele frequency without DN H1, 0.04/H2, 0.96 vs with DN H1, 0.05/H2, 0.95; P=0.551). Conclusions: The polymorphism in the gene for the atrial natriuretic peptide does not seem to play a major role in the development of diabetic nephropathy in either type I or in type II diabetes.     

Association of uromodulin gene polymorphism with diabetic kidney disease in Han population in Tianjin of China

Objective To investigate the association of single nucleotide polymorphism (SNP) rs13333226 in uromodulin (UMOD) gene with diabetic kidney diseases (DKD) in Han population in Tianjin, China. Methods A total of 210 type 2 diabetes (T2DM), 90 normal controls (NC) and 280 DKD patients were recruited. According to the level of estimated glomerular filtration rate (eGFR), the DKD subjects were further subdivided into three groups: GFR≥90 ml/min group (n=105), 60 ml/mim≤GFR＜90 ml/min group (n=84) and GFR＜60 ml/min group (n=91). Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for UMOD rs13333226C genotyping. Results The frequencies of AA, GA, GG genotype were 27.8%, 58.9%, 13.3% in NC group and 41.0%, 48.6%, 10.5% in T2DM group and 54.3%, 36.1%, 9.6% in DKD group．The frequency of G allele was 42.8% in NC group, 34.8% in T2DM group and 27.7% in DKD group. The genotype distribution of UMOD was statistically significant between NC group and DKD group, and between T2DM group and DKD group (P＜0.05). G allele of UMOD was an independent protective gene polymorphism of DKD in Logistic regression (B=-0.248, Wald=8.012, P=0.021, OR=0.780, 95%CI 0.612-0.968). Conclusion The G allele of UMOD gene may be an independent protective factor of DKD in Han population in Tianjin, China.