MDR基因多态性对肾移植受者环孢素A急性肾毒性发生率的影响

目的 探讨供、受者细胞膜P-糖蛋白(P-gp)的编码基因(MDR基因)多态性对肾移植受者环孢素A(CsA)急性肾毒性发生率的影响.方法 采用聚合酶链反应/限制性片段长度多态性(PCR/RFLP)方法分析173例肾移植供、受者MDR基因外显子26(exon26)基因型;分别以供者和受者exon26基因型进行分组,观测术后3个月内受者血尿素氮、血肌酐和24 h尿量等指标与供、受者exon26基因型之间的关系.结果 供者和受者exon26基因型均为CC、CT和TT三种类型.173例供者中,CC、CT和TTr型比例分别为:28.3%(49/173)、41.6%(72/173)和30.0%(52/173);173例受者中,CC、CT和TT基因型比例分别为:27.7%(48/173)、40.5%(70/173)和31.8%(55/173).供、受者间exon26基因型的类型和分布比例相似,差异无统计学意义(P>0.05).肾移植术后,共有48例受者符合CsA急性肾毒性诊断标准.按受者CC、CT、TT型分组时,各组受者CsA急性肾毒性的发生率分别为25.0%(12/48)、41.7%(20/48)和33.3%(16/48),各组间比较,差异无统计学意义(P>0.05);按供者CC、CT和TT基因型分组时,各组受者术后CsA急性肾毒性的发生率分别为14.6%(7/48)、50.0%(24/48)和35.4%(17/48),CC与CT和TT型之间比较,差异有统计学意义(P<0.05).而且.受者CC型组肾功能异常发生率明显高于供者CC型组(P<0.05).结论 MDR基因多态性对肾移植受者CsA急性肾毒性发生率具有明显的影响,而发挥作用是供者MDR基因多态性.     

肾移植患者的多药耐药基因外显子26基因型与术后他克莫司用量的关系

目的研究肾移植患者的多药耐药基因(MDR1)外显子26(exon26)的基因型与术后他克莫司(FK506)用量的关系。方法回顾106例肾移植术后常规使用FK506患者的临床资料。肾移植患者MDR1 exon26基因分型的方法为:提取患者的DNA,采用聚合酶链反应(PCR)扩增MDR1基因,检测限制性内切酶片段的多态性(RFLP)。根据MDR1 exon26基因分型将患者分为CC、CT和TT 3组。检测各组患者肾移植后第3、6和12个月的FK506血药浓度,比较各组患者FK506血药浓度／FK506用量(μg·L-1／mg·kg-1·d-1)的比值及术后1个月内的急性排斥反应发生率。结果受者经MDR1 exon26基因分型示:CC型32例(30．2％),TT型30例(28．3％),CT型44例(41．5％)。CC型患者FK506血药浓度／FK506用量的比值明显低于CT型和TT型(P<0．01),而CT型患者又低于TT型(P<0．05)。CC型患者的排斥反应发生率明显高于CT和TT型(P<0．05),CT与TT型比较,差异无统计学意义(P>0．05)。结论MDR1 exon26 CC型的患者与CT或TT型比较,需服用更高剂量的FK506才能取得与CT或TT型相似的血药浓度。因此,了解患者的MDR1 exon26基因型有利于指导患者肾移植术后个体化用药。     

乳腺癌MDR1基因多态性与其表达及化疗血液毒性的关系

目的：探讨乳腺癌多药耐药基因MDR1基因多态性与其表达及与化疗血液毒性之间的关系。 方法：提取92例乳腺癌患者外周静脉血DNA,利用限制性片段长度多态性（PCR-RFLP）技术检测MDR1外显子（exon）26（C3435T）位点的多态性,并用RT-qPCR方法检测该92例患者癌组织及其中26例配对癌旁组织MDR1 mRNA的表达。根据患者的临床资料,分析C3435T位点多态性与化疗血液学毒性的关系。 结果：92例乳腺癌中, C3435T位点CC,CT,TT 3种基因型分别占21.7%（20/92）,62.0%（57/92）和16.3%（15/92）,所有乳癌组织中均有不同程度的MDR1 mRNA表达,但各基因型间MDR1 mRNA表达水平差异无统计学意义（F=0.173,P=0.841）;癌组织MDR1 mRNA表达水平明显高于其对应的癌旁组织[（3.83±5.27） vs. (1.81±4.42),t=2.522,P=0.018]。C3435T各基因型患者中性粒细胞减少反应差异有统计学意义,CC型中性粒细胞减少（III~IV度）的发生频率（5.0%）明显低于CT和TT型（26.3%和46.7%;χ2=8.075,P=0.018,95%CI=0.017~0.022）,而白细胞减少、贫血、血小板减少等方面各基因型间无统计学差异（均P>0.05）。 结论：乳腺癌患者MDR1基因C3435T位点多态性与MDR1基因表达水平无明显关系,CT和TT型患者化疗后发生中性粒细胞减少症的风险较大。     

冠心病与内皮型一氧化氮合酶基因多态性的关系

目的 探讨内皮型一氧化氮合酶(eNOS)基因-786T/C,4a4b,894G/T等3个多态性位点与冠心病(CAD)发病相关.方法 对146例中国汉族人群CAD患者和113例正常对照进行遗传学分析,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)和PCR技术分析2个SNP位点即-786T/C和894G/T,以及1个VNTR位点4a4b,检测各位点基因型和等位基因频率,采用HaploView 4.0及SPSS 13.0软件经x2检验比较两组间各位点基因型及等位基因频率的差异.结果 CAD组中eNOS基因-786T/C位点CC基因型频率为2.0%,4a4b位点4a/4a基因型频率为5.4%,对照组eNOS基因-786T/C位点CC基因型频率为0.0%,4a4b位点4a/4a基因型频率为0.9%,差异有统计学意义(P＜0.05).CAD组和对照组在eNOS基因的894G/T位点等位基因和基因型频率分布差异均无统计学意义(P＞0.05).结论 eNOS基因-786T/C和4a4b多态性与中国汉族人群CAD存在关联,C等位基因和4a等位基因可能是CAD发病的危险因素.eNOS基因894G/T位点与CAD发病无明显相关.

Abstract：

Objective To investigate the relationship between the 3 polymorphisms ( -786T/C,4a4b,894G/T) in endothelial nitric oxide synthase (eNOS) gene and coronary artery disease (CAD).Methods 146 patients with CAD and 113 healthy unrelated individuals in a Chinese Han nation were involved.The genotype and allele frequency of each polymorphism of the eNOS gene in these patients and normal controls were examined by using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) or PCR methods.Genotypes and allele frequency were analyzed by HaploView 4.0 and SPSS13.0 software.Results The frequency of CC genotype of the -786T/C was 2.0%,and that of 4a/4a genotype of the 4a4b was 5.4% in CAD.The frequency of CC genotype of the - 786T/C was 0.0%,and that of 4a/4a genotype of the 4a4b was 0.9% in controls ( P＜0.05 ).There were significant differences in both allele and genotype frequency of -786T/C and 4a4b between CDA group and control group.Between patients with CAD and controls,there were no significant differences in the frequency of the genotypes and alleles of the 894G/T in eNOS gene.Conclusion The - 786T/C and 4a4b polymorphisms of eNOS gene may be associated with CAD.The individuals with C allele of - 786T/C and 4a allele of 4a4b are susceptible to CAD.There is no significant correlation between 894G/T polymorphism in eNOS gene and CAD.     

食管癌易感性与白细胞介素-8基因多态性的关系

目的 探讨白细胞介素(IL)-8多态性与食管癌发生危险因素的关系.方法 采用以医院为基础的病例对照研究方法,以高分辨率熔解曲线(HRM)小片段扩增子法分析351例食管癌和384例非肿瘤对照IL-8基因多态性,计算各种基因型的食管癌发生风险及其95%可信区间.结果 IL-8-251(A/T)多态3种基因型TT、AT、AA在食管癌组和非肿瘤对照组的频率分别为39.9%(IT)、54.7%(AT)、5.4%(AA)和43.8%(TT)、45.3%(AT)、10.9%(AA),与携带IL-8-251TT基因型的个体比较,IL-8-251 AA等位基因型可以减少46%食管癌的发病风险.结论 IL-8-25l(A/T)基因多态性AA基因型可能是食管癌发生的保护性因素.

Abstract：

Objective To investigate the relationship between interleukin-8 (IL-8) polymorphism and the susceptibility to esophageal cancer in a Chinese Han population. Methods Genotypes were determined by the high resolution melting (HRM) method in 351 esophageal cancer cases and 384 controls without cancers. Results The IL-8 genotype frequency was 39. 9% ( TF), 54. 7% (AT), 5.4% (AA)in the esophageal cancer group, and 43.8% (TT), 45. 3% (AT), 10. 9% (AA) in the control group respectively. Logistic regression analyses revealed that the risk associated with IL-8 variant genotype was 0. 54 (95% CI =0. 30-0. 98) for IL-8 AA compared with its wild-type homozygote. Conclusion IL-8 AA polymorphism may serve as a protective biomarker of esophageal cancer susceptibility.     

原发性胆囊癌胆囊黏膜和胆汁中螺杆菌相关基因的检测

目的 探讨人类原发性胆囊癌(primary carcinoma of the gallbladder,PCG)的发生是否与幽门螺杆菌(Helicobacter pylor,Hp)的感染有关,为原发性胆囊癌的治疗提供新的思路.方法 选取18例PCG患者、40例慢性胆囊炎、胆囊结石(chronic cholecystitis or cholelithiasis,CC)患者以及20例排除PCG和CC的患者(对照组)手术后的胆囊黏膜、胆汁,采用巢式PCR方法扩增螺杆菌特异的16S rRNA基因.阳性者检测Hp特异26 KDa蛋白1基因和4个相关功能基因(cagA、vacA、rps4、ureA),并进行测序分析,确定检出的细菌是否为Hp.结果 PCG组胆囊黏膜和胆汁标本有29份检出螺杆菌属16S rRNA基因,阳性率为81%,显著高于CC组(50%)和对照组(20%).PCG 组螺杆菌属16S rRNA基因阳性29份标本中,15份cagA基因阳性,阳性率为52%;25份26 KDa基因阳性,阳性率为86%;14份ureA基因阳性,阳性率为48%;未扩增出vacA和rps4基因;cagA、26 KDa、vacA、ureA和rps4基因至少一项阳性的样本数为27份,阳性率为93%.经测序鉴定,检测出的螺杆菌与Hp有99%以上的同源性.结论 PCG患者胆囊黏膜和胆汁中存在Hp感染,且感染率高.初步提示胆道系统Hp感染与PCG有关.

Abstract：

Objective To investigate the relationship between helicobacter pylor (Hp) and primary carcinoma of the gallbladder (PCG), providing a theoretical basis for the treatment of PCG.Methods Mucosa and bile of gallbladder samples were collected from 18 patients with PCG (PCG group), 40 patients with chronic cholecystitis or cholelithiasis (CC group), and 20 patients with no PCG and CC (control group). 16S rDNA-based polymerase chain reaction (PCR) followed by DNA sequence analysis of the obtained PCR fragments was performed. A developed search for Hp was also carried out by PCR. Five genes specific for Hp were amplified. Results In the PCG group, 81% samples of mucosa and bile were positive for Helicobacter-specific 16S rRNA gene. The positive rate in PCG group was significantly higher than those of the CC group (50%) and the control group (20%).Of the 16S rDNA sequence of Helicobacter pylori positive samples of mucosa and bile, 52% of samples were positive in cagA, 86% were positive in 26 KDa, 48% were positive in ureA separately. The vacA and rps4 genes were never detected in any of the samples of mucosa and bile. At least one gene was positive in 93% of samples of musca and bile. The sequencing showed more than 99% homology of Helicobacter 16S rRNA in PCG and Hp groups. Conclusion A higher infection rate is present in bile and gallbladder mucosa from patients with PCG. Hp in the biliary system may be one of the risk factors for occurrence of PCG.     

汉族与维吾尔族尿石症患者血管内皮生长因子基因多态性研究

目的探讨不同种族之间血管内皮生长因子（VEGF）基因型频率的分布。方法应用聚合酶链反应一限制性片段长度多态性分析术（PCR—RFLP）、基因测序等技术检测汉族尿石症患者、健康人群以及维吾尔族尿石症患者和健康人群各200名的VEGF基因多态性,比较两组VEGF基因型和等位基因的分布频率。结果汉族尿石症患者中CC基因型占1．00％、CT基因型占80．50％、TT基因型占18．50％,而维吾尔族尿石症患者中CC基因型占1．50％、CT基因型占69．50％、TT基因型占29．00％。汉族健康者中CC基因型占0．00％、CT基因型占85．50％、TT基因型占14．50％,维吾尔族健康者中CC基因型占0．25％、CT基因型占85．50％、TT基因型占13．75％。两民族VEGF基因型的分布频率差异有显著性（P〈0．05）。结论汉族、维吾尔族尿石症患者VEGF基因多态性分布频率有差异。     

多药耐药基因多态性与免疫抑制剂应用关系的研究

目的探讨多药耐药基因(MDR1)外显子21(exon21)的基因多态性对肾移植术后患者免疫抑制剂应用的影响. 方法选择同种异体肾移植术后患者168例.采用聚合酶链反应(PCR)扩增MDR1基因,限制性内切片段多态性(RFLP)方法对MDR1基因进行分型.根据分型将患者分为GG、GT和TT 3组,对肾移植术后第1、3、6、12个月中每月每2组患者间的CsA浓度与每天每公斤体重的CsA剂量比值进行比较. 结果168例患者中GG型46例(27.4%),GT型76例(45.2%),TT型46例(27.4%).GG型患者CsA浓度剂量比值明显低于GT型和TT型患者,差异均有统计学意义(P＜0.01),而GT型患者CsA浓度剂量比值又低于TT型患者,差异有统计学意义(P＜0.05).结论同种异体肾移植患者MDR1exon21基因型和CsA血药浓度与每天每公斤体重CsA剂量的比值有明显关系.GG型患者的比值明显低于GT或TT型患者,GG型患者达到相似的血药浓度需服用更高剂量的CsA.     

雌激素受体α基因Pvu Ⅱ位点多态性与女性膝骨关节炎相关性的Meta分析

[目的]探讨雌激素受体α(ERα)基因PvuⅡ位点多态性与女性膝骨关节炎(OA)易感性的关系。[方法]检索Pub Med数据库、web of science数据库、万方数据库、中国知网全文数据库、维普数据库、中国生物医学文献数据库发表的有关雌激素受体α(ERα)基因PvuⅡ位点多态性与女性膝骨关节炎的病例对照研究文献,OR值及95%CI为效应指标,采用固定或随机效应模型进行合并分析,并进行偏倚评估。应用Revman 5.1软件进行统计学处理。[结果]共纳入文献5篇,包括1 414例骨关节炎患者,903例健康对照者。PvuⅡ位点总人群等位基因及基因型(CT vs TT;CC vs TT;CT+CC vs TT;CC vs CT+TT)合并OR值均1;按地区分层提示亚洲地区人群等位基因及基因型(CC vs TT;CT+CC vs TT;CC vs CT+TT)合并OR值均1,欧美地区人群等位基因及基因型(CT vs TT;CC vs TT;CT+CC vs TT;CC vs CT+TT)合并OR值均1。另外,亚洲地区女性人群CC vs TT及CC vs CT+TT基因型P0.05(P=0.04;P=0.02)差异有统计学意义。[结论]雌激素受体α(ERα)基因PvuⅡ位点多态性与亚洲女性膝骨关节炎遗传易感性有一定关联性。     

亚甲基四氢叶酸还原酶基因C677T多态性与男性不育的相关性及机制研究

目的探讨亚甲基四氢叶酸还原酶(MTHFR)基因C677T基因多态性与男性不育人群精液质量指标的相关性及可能机制。方法回顾性分析我院2016年1月至2017年3月就诊的59名男性不育症患者的临床资料,根据MTHFR基因型C677T分型分为CC、CT和TT基因型3组,比较各组精子质量参数,包括各组重度少、弱精子症、严重少弱精子症和正常精子者的比例,精子存活率、DNA完整性和顶体完整性,同时分析各组精液活性氧(ROS)水平。结果 CT基因型组正常精子者比例显著低于CC基因型组(P0.05);TT基因型组重度少、弱精子症、严重少弱精子症比例显著高于CC基因型组,严重少弱精子症比例亦显著高于CT型组,正常精子比例显著低于CC型和CT型组(P均0.05)。CT基因型组精子存活率显著低于CC基因型组,TT基因型组精子存活率显著低于CC、CT基因型组(P均0.05)。TT基因型组较CC、CT基因型组精子DNA完整性相关指标均有显著升高(P0.05);TT基因型组精子顶体完整性则显著低于CC、CT基因型组(P0.05)。CT基因型组ROS水平显著高于CC基因型组(P0.05),TT基因型组ROS水平显著高于CC、CT基因型组(P0.05)。结论 MTHFR基因C677T基因多态性与男性不育人群的精子质量指标相关,可能与ROS水平增高有关。     

Association between ABCB1 (MDR1) gene 3435 C>T polymorphism and colchicine unresponsiveness of FMF patients

The multidrug resistance gene-1 (MDR1, adenosine triphosphate-binding cassette transporter: ABCB1, P-glycoprotein) encodes membrane proteins that play a crucial role in protecting cells from xenobiotics, chemicals, and drugs. The TT genotype of 3435 codon in exon 26 of MDR1 gene causes overexpression of gene activity and effluxes many chemically diverse compounds across the plasma membrane. We studied the association between C3435T polymorphisms (single nucleotide polymorphism) of MDR1 gene and colchicine-resistant familial Mediterranean fever (FMF) patients. Total genomic DNA samples from 52 FMF patients of colchicine unresponsiveness were used for FMF (MEFV) and MDR1 genes profile analyses. Target genes were genotyped by multiplex PCR-based reverse-hybridization Strip Assay method. The preliminary current results showed increased T allele frequency (0.596) in colchicine unresponsiveness of FMF patients. The distributions of the CC, CT, and TT genotypes in colchicine nonresponder FMF patients were 17%, 46%, and 37%, respectively. Our results indicate that C3435T polymorphism in exon 26 of MDR1 gene is associated with colchicine resistance in nonresponder FMF patients during the common therapy protocol.     

MDR-1 C3435T polymorphism influences cyclosporine a dose requirement in liver-transplant recipients

BACKGROUND: Cyclosporine A (CsA) is characterized by high interindividual variations in oral bioavailability and a narrow therapeutic index. CsA is a substrate for P-glycoprotein, a member of the ABC transporter family encoded by the multiple drug-resistant gene MDR1. METHODS: Because MDR1 gene exon 26 C3435T polymorphism influences intestinal P-glycoprotein expression, we investigated whether this polymorphism was correlated with variation in CsA dose requirement and concentration/dose ratio in 44 liver-transplant recipients during 1 month after transplantation. CsA concentration was measured 2 hours after administration (C2), according to international recommendations. RESULTS: The MDR-1 wild-type genotype (3435CC) was observed in 15 patients (34%), whereas 21 (48%) patients were heterozygous (3435CT), and 8 (18%) patients were homozygous for the mutation (3435TT). There was no significant difference between the three groups regarding corticosteroids treatment or renal function during this period. One to 3 days after liver transplantation, when every patient received a similar CsA weight-adjusted dose, the concentration/dose ratio was correlated with exon 26 single nucleotide polymorphism and was significantly higher in subjects homozygous for the mutation (P=0.012). This was confirmed 1 month after transplantation (P=0.049), when the dose was adjusted to maintain the C2 target level of 1,000 microg/L and we observed that TT patients required approximately 50% lower weight-adjusted CsA dose than wild-type patients (P=0,033). CONCLUSIONS: These findings demonstrate that the MDR1 exon 26 C3435T polymorphism is a major determinant of CsA concentration/dose ratio in liver-transplant recipients and is predictive of the dose of CsA to be administered to achieve the target C(2) concentration.     

Association between ABCB1 (MDR1) Gene 3435 C>T Polymorphism and Colchicine Unresponsiveness of FMF Patients

The multidrug resistance gene-1 (MDR1, adenosine triphosphate-binding cassette transporter: ABCB1, P-glycoprotein) encodes membrane proteins that play a crucial role in protecting cells from xenobiotics, chemicals, and drugs. The TT genotype of 3435 codon in exon 26 of MDR1 gene causes overexpression of gene activity and effluxes many chemically diverse compounds across the plasma membrane. We studied the association between C3435T polymorphisms (single nucleotide polymorphism) of MDR1 gene and colchicine-resistant familial Mediterranean fever (FMF) patients. Total genomic DNA samples from 52 FMF patients of colchicine unresponsiveness were used for FMF (MEFV) and MDR1 genes profile analyses. Target genes were genotyped by multiplex PCR-based reverse-hybridization Strip Assay method. The preliminary current results showed increased T allele frequency (0.596) in colchicine unresponsiveness of FMF patients. The distributions of the CC, CT, and TT genotypes in colchicine nonresponder FMF patients were 17%, 46%, and 37%, respectively. Our results indicate that C3435T polymorphism in exon 26 of MDR1 gene is associated with colchicine resistance in nonresponder FMF patients during the common therapy protocol.     

Association of cadherin23 single nucleotide polymorphism with age-related hearing impairment in han chinese

Objective Genetic variation of cadheri23 (cdh23; 753G>A in exon 7) has been implicated with age-related hearing impairment (ARHI) in mice. This study aimed to test the association of the CDH23 tag single nucleotide polymorphism (SNP) in intron 7 with ARHI in Han Chinese. Study Design Individual cohort study. Setting Tertiary medical center. Subjects and Methods A total of 1175 Han Chinese subjects were divided into the case group (n = 310, 26% with poorest hearing) and the control group (n = 308, the 26% with best hearing) according to the Z(high) score converted from the original frequency-specific hearing thresholds. The CDH23 SNP locus (rs7087735: C/T) in intron 7 (coordinate: 72996763) shown in the HapMap was genotyped with correlation to the hearing phenotype. Results The genotype distributions of CDH23 (CC/CT/TT) were not significantly different between the case and control group (P = .489). Compared with genotype CC, the odds ratios of the genotypes CT and TT for ARHI were not significantly different after adjustment for other environmental factors (P = .299 for CT; P = .610 for TT). Conclusions Despite that the Ahl allele of Cdh23 had been implicated with ARHI in mice, we found no positive association of the CDH23 tag SNP in intron 7 with ARHI in Han Chinese.     

过氧化物酶体增殖物激活受体γC161T基因多态性与糖皮质激素性骨质疏松症的关系

目的 探讨中国人过氧化物酶体增殖物激活受体γ(PPARγ)基因外显子6 C161T多态性与糖皮质激素性骨质疏松症(GIO)的相关关系。方法 应用聚合酶链反应-限制性片段长度多态性(PCR-RELP)方法测定208例正常健康人（Ⅰ组）、168例非GIO患者（Ⅱ组）和104例GIO患者（Ⅲ组）PPARγ基因外显子6 C161T的基因型。应用双能X线骨密度仪(DEXA)测定股骨、腰椎等部位的骨密度。 结果 外显子6 C161T有CC、CT、TT 3种基因型。GIO组CC基因型频率显著低于正常对照组；CT和TT基因型频率显著高于正常对照组。非GIO组、应用激素组（GIO组＋非GIO组）与正常对照组比较，各基因型频率差异均无统计学意义。正常对照组C161T的CC基因型组各部位的骨密度有高于CT和TT基因型组的趋势，但差异无统计学意义。非GIO组和GIO组C161T的CC基因型组腰椎的骨密度明显高于CT和TT基因型组 (P ＜ 0.05)，分别为非GIO组CC型（1.04±0.17） g/cm2，CT+TT型（1.02±0.07） g/cm2；GIO组CC型（0.94±0.12） g/cm2，CT+TT型（0.83±0.08） g/cm2。经年龄、体重指数等因素校正后，差异仍有统计学意义(P ＜ 0.05）。 结论 PPARγ基因C161T基因型在正常人和应用激素患者之间无明显差异，它可能与肾小球肾炎的发病无关。C161T基因型在GIO组和正常对照组之间差异有统计学意义，它可能与糖皮质激素性骨质疏松症的发病有关。PPARγ基因C161T多态性与应用糖皮质激素患者腰椎的骨密度有关。等位基因C可能是骨量的保护因子，它可能与应用糖皮质激素后骨量的丢失有关。     

霉酚酸酯药代动力学与多重耐药基因1多态性的相关性研究

目的 研究肾移植受者术后早期霉酚酸酯(MMF)的药代动力学与人类多重耐药基因1(MDRI)多态性的相关性.方法 选择初次肾移植的汉族受者28例,肾移植术后2周时,于口服MMF之前及服药后0.5、1、1.5、2、4、6、8、10、12 h共10个时间点分别采集外周血,以高效液相色谱(HPLC)法测定全血霉酚酸酯(MMF)的活性成分霉酚酸(MPA)的浓度,直接观察其峰值浓度(Cmax)和达峰时间(Tmax).应用Winnolin 3.1软件计算MPA 0～12 h药物时间一浓度曲线下面积(AUC)和平均滞留时间(MRT).同时从外周血提取基因组DNA,应用多聚酶链反应-限制性片断长度多态性(PCR-RFLP)测定MDR1第12、21、26号外显子C1236 T、G2677 T/A、C3435 T单核苷酸多态性(SNP).比较3个SNP位点的不同基因型、单倍型间MMF药代动力学参数的差异;比较MPA高暴露组(MPA AUC≥60 mg·h-1·L-1)与MPA低暴露组(MPA AUC＜60 mg·h-1·L-1)间MDR1多态性差异.结果 MDR1第12、21、26号外显子SNP位点突变型纯合子基因型(1236 TT、2677 TT/AA、3435 TT)频率分别为0.368、0.184和0.211.1236 TT基因型受者MPA AUC水平显著高于1236 cc/CT受者,分别为(65.36±11.51)mg·h-1·L-1和(53.33±13.77)mg·h-1·L-1(P=0.032).MPA高暴露组第12号外显子SNP位点上,TT基因型频率显著高于低暴露组,分别为66.7%和15.8%(P=0.013,OR=2.526);T等位基因频率有高于低暴露组的趋势,分别为83.3%和53.3%(P=0.072).结论 具有MDR1第12号外显子TT等位基因的受者,肾移植早期MPA AUC显著高于同一位点其他基因型受者,是MPA高暴露的危险个体.     

Association of transforming growth factor-beta (TGF-beta) T869C (Leu 10Pro) gene polymorphisms with type 2 diabetic nephropathy in Chinese

INTRODUCTION: Transforming growth factor-beta (TGF-beta) is known to play a pivotal role in the regulation of extracellular matrix (ECM) accumulation. Since diabetic nephropathy (DMN) is characterized by basement membrane thickening and mesangial expansion, control of ECM deposition is believed to be important in the pathogenesis of the disease. Recently, TGF-beta T869C (Leu 10Pro) gene polymorphism has been identified which may be associated with circulating TGF-beta levels. METHODS: In order to examine the relationship between TGF-beta gene polymorphism with DMN in Chinese, we carried out a case-control study, which recruited 123 Chinese type 2 diabetic patients with an average duration of diabetes for 12 years. A total of 58 patients who developed DMN (micro- or macroalbuminuria, with or without renal impairment) were compared with 65 diabetic patients without DMN despite similar duration of disease (normoalbuminuric and creatinine <120 micromol/L). TGF-beta T869C (Leu 10Pro) gene polymorphism was determined by polymerase chain reaction (PCR). RESULTS: Both groups of patients had similar baseline characteristics, including blood pressure, diabetic control, and duration of diabetes. Distribution of TGF-beta T869C (Leu 10Pro) genotype among the whole group is confined to Hardy Weinberg equilibrium. The DMN+ group has higher frequency of TGF-beta CC/CT genotypes than the DMN- group [CC, CT, TT = (DMN+) 46, 45, 9 (%) vs. (DMN-) 37, 37, 26 (%), P < 0.05]. C allele frequency is also higher in the DMN+ group than DMN- group (69% vs. 55%, P < 0.05). The adjusted odds ratio for TGF-beta CC/CT vs. TT genotype to develop DMN is 3.8 (3.2 to 4.4). Multivariate logistic regression analysis [hypertension, gender, age, duration of diabetes, hemoglobin (HbA1c), usage of angiotensin-converting enzyme (ACE) inhibitor, and cholesterol level] showed that TGF-beta genotype (P = 0.03) is an independent predictor for type 2 DMN. Among patients with DMN, those with TGF-beta CC/CT genotypes also had worse renal function and increased risk for macroalbuminuria. CONCLUSION: Our results suggest that TGF-beta T869C (Leu 10Pro) gene polymorphism is associated with DMN in Chinese.     

Association of transforming growth factor beta1 genotype with therapeutic response to active vitamin D for postmenopausal osteoporosis.

Transforming growth factor beta (TGF-beta) is an important regulator of bone metabolism, its effects being intertwined with those of estrogen and vitamin D. A T-->C polymorphism in exon 1 of the TGF-beta1 gene, which results in the substitution of proline for leucine, is associated with bone mineral density (BMD). However, it is not known whether this polymorphism affects the response to treatment with active vitamin D or to hormone replacement therapy (HRT) in individuals with osteoporosis. Changes in BMD at the lumbar spine (L2-L4 BMD) were compared among TGF-beta1 genotypes in 363 postmenopausal Japanese women who were divided into three groups: an untreated, control group (n = 130), an active vitamin D treatment group (n = 117), and an HRT group (n = 116). TGF-beta1 genotype was determined with an allele-specific polymerase chain reaction assay. In the control group, the rate of bone loss decreased according to the rank order of genotypes TT (homozygous for the T allele) > TC (heterozygous) > CC (homozygous for the C allele), with a significant difference detected between the CC and TT genotypes. The positive response of L2-L4 BMD to HRT increased according to the rank order of genotypes TT < TC < CC, although the differences among genotypes were not statistically significant. Individuals with the CC genotype responded to active vitamin D treatment with an annual increase in L2-L4 BMD of 1.6%, whereas those with the TT or TC genotypes similarly treated lost bone to a similar extent as did untreated subjects of the corresponding genotype. These results suggest that TGF-beta1 genotype is associated with both the rate of bone loss and the response to active vitamin D treatment.