翻译

有关基因的资料

目的：本研究的目的是确定是否血管紧张素转换酶抑制剂的插入/缺失（I/D），血管紧张素（RSA）的相关性。 研究设计：共251例不明原因流产，和126名健康对照者中至少有一个活产无流产史。、 结果：比值比（OR）的ACE ID (OR = 2.423；95%可信区间（CI）= 1.417-4.142; p = 0.001）和ACE II（OR= 2.050；95% CI = 1.143–3.675；P = 0.018）的ACE DD基因型之间差异极显著例特发性RSA与对照组无显著性差异；然而，患者和对照组之间就AT1R 1166a＞C和AGT M235 III类一致T多态性有。在一个单倍型分析I-A（ P = 0.010），D-A (p = 0.004), I-A-T (p = 0.033), D-A-T (p = 0.0005), 和 D-C-T (p = 0.013)和协同效应在患者和对照组的方法，导出多态性对具有显著效果。

结论：本研究表明，ACE，AT1R和AGT基因多态性和单倍型为在韩国女性特发性RSA的风险的遗传因素。

The renin–angiotensin–aldosterone system (RAAS) or therenin–angiotensin system (RAS), which is one of the important factors controlling blood pressure, volume in the cardiovascular system, and electrolyte balance is a hormone system. 肾素血管紧张素醛固酮系统（RAAS––）或肾素-–血管紧张素系统（RAS）

，这是一个控制血压的重要因素，在心血管系统的体积，和电解质的平衡是一种激素系统。

Angiotensinogen (AGT), which is produced by the liver, is part of the renin–angiotensin system and is a major target for drugs that lower blood pressure. AGT is also known as a renin . Angiotensin I is produced by the action of rennin on angiotensinogen. Angiotensin I is catalyzed by angiotensin I converting enzyme (ACE), which converts angiotensin I to angiotensin II, a potent vasoconstrictor in the cardiovascular system.

血管紧张素原（AGT），它是由肝脏产生的，是肾素血管紧张素系统的一部分，是–降血压药物的主要目标。AGT也被称为肾素底物。血管紧张素是由肾素-血管紧张素作用产生的。血管紧张素1是由血管紧张素1转换酶（ACE），将血管紧张素1转换为血管紧张素II，在心血管系统中一种有效的血管收缩剂。

Moreover, ACE degrades bradykinin, a potent vasodilator, and other vasoactive peptides. Angiotensin II has vasopressor（血管加压素）effects and regulates aldosterone secretion. Angiotensin II functions through at least two types of receptors (angiotensin II type 1 receptor

[AT1R] and angiotensin II type 2receptor [AT2R]).

AT1R has vasopressor effects and regulates aldosterone secretion. AT1R mediates the major cardiovascular effects, including vasoconstriction, aldosterone synthesis and secretion, increased vasopressin secretion, vascular smooth muscle cell proliferation, decreased renal blood flow, renal renin inhibition, renal tubular sodium reuptake, modulation of central sympathetic nervous system activity, and cardiac contractility [2].

AT1RAT1R介导的心血管作用，包括血管收缩，醛固酮的合成与分泌，加压素分泌增加，血管平滑肌细胞增生，肾血流量减少，肾肾素抑制，肾小管对钠的重吸收，中枢交感神经系统活动和心肌收缩能力的

有关基因的资料

调制[ 2 ]。

Angiotensin II has prothrombotic potential through adhesion and aggregation of platelets and production of plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator inhibitor-2 (PAI-2),which are significant components of the fibrinolysis cascade. AT1R and AT2R are expressed in human embryos during pregnancy and are included in organ development.

血管紧张素II有形成血栓的潜力，通过血小板粘附和聚集作用，血浆纤溶酶原激活物抑制剂-1（PAI-1）聚合和纤溶酶原激活物抑制物-2（PAI-2）的生产，并且这是纤溶级联的重要组成部分。AT1R和AT2R的表达在人类胚胎在妊娠过程中，包括在器官发育阶段。

ACE, AT1R, and AGT gene polymorphisms have been reported to be associated with abortion

[3–5]. There genes are also related to various vascular diseases, such as hypertension, coronary artery disease, ischemic stroke, cardiovascular disease, and pregnancy-related diseases, such as pregnancy-induced hypertension (pre-eclampsia) or serious pregnancy complications [6,7].

ACE，AT1R，和AGT基因多态性已被报道与堕胎相关[ 3 –5]。有基因也与多种血管性疾病有关，如高血压、冠状动脉疾病、缺血性中风、心血管疾病，与妊娠有关的疾病，如妊娠高血压综合征（先兆子痫）或严重的妊娠并发症[6,7]。

are many studies pertaining to ACE I/D, AT1R1166A>C, and AGT M235T polymorphisms, this is the first report which evaluates these three genes in idiopathic RSA by using haplotype-based analyses. We therefore determined the association between ACE I/D, AT1R 1166A>C, and AGT M235T polymorphisms in Korean women with idiopathic RSAs.

2. Materials and methods

2。材料与方法

2.1. Patients

The patient group was comprised of 251 Korean women (age range; 22–45 years, mean age SD：31.97±4.30 years, BMI：21.58±3.58) who were diagnosed with at least three consecutive spontaneous abortions. The patients were enrolled between March 1999 and February 2008 in the Department of Obstetrics and Gynecology, and the Fertility Center of CHA Bundang Medical Center in Seongnam, South Korea. Patients with a pregnancy loss were identified by hCG testing, ultrasound, and/or physical examination (<20 weeks gestational age).

患者组由251个韩国女性（年龄范围；22–45岁，平均年龄4.30年，SD；BMI；31.97，21.58 3.58）确诊为至少三个连续自然流产。患者1999年3月和二月的2008间产科病区，妇科，和茶Bundang医疗中心在城南的生育中心，韩国。有一个流产患者进行绒毛膜促性腺激素检测，超声检查，和/或体检（＜20孕周）。

Women who had previous venous or arterial thromboembolism, an indication for anticoagulant

有关基因的资料

treatment during pregnancy, endocrine disorders (such as diabetes mellitus or untreated thyroid dysfunction), and anatomic, chromosomal, infectious, or autoimmune causes of spontaneous abortion were excluded from the study group. 妇女谁了以前的静脉或动脉血栓栓塞，怀孕期间抗凝治疗迹象，内分泌失调（如糖尿病或甲状腺功能异常，未经处理的）和解剖，染色体，传染，或免疫性原因引起的自然流产被排除在研究组之外。

Also, we excluded patients who had a family history of RSA and spousal chromosomal abnormalities. 4.0% of the patients were experienced gravidity, and 10.4% had a parity history. The control group consisted of 126 women with at least 1 live birth and no history of pregnancy loss (age range; 23–43 years, mean age SD; 31.22±4.30 years，BMI：21.72±3.40).

另外，我们排除了那些有RSA家族史和配偶染色体异常的患者。4%的患者经历了妊娠，10.4% 的历史。对照组包括126名至少有1个活产和无流产史妇女（年龄范围；23–43岁，平均年龄SD：31.22±4.30 岁，BMI：21.72 ±3.40）。

Medical histories were obtained, and physical examinations, laboratory tests, and ultrasonography were performed on the women in the control group. The Institutional Review Board of CHA Bundang Medical Center approved the study in 1999, and all subjects gave written informed consent.

得到对照组妇女的病史和对她们进行体格检查，实验室检查，超声检查。1999年CHA Bundang医疗中心的机构审查委员会批准此研究，并所有受试者知情同意书上签过字儿。

Chromosomal analyses of blood and/or abortus samples were carried out using standard protocols [8,9,12,13]. Metaphase chromosomes were stained using the GTG-banding method and20 metaphases per sample were analyzed.

血液和/或流产标本的染色体分析使用标准协议进行的[8,9,12,13 ] 。中期染色体进行染色，使用GTG显带法和中期每样品进行分析。

2.2. Genetic analysis of ACE, AT1R and AGT polymorphisms

ACE insertion or deletion was screened by a polymerase chain reaction (PCR) using a sense primer (5’-CTG GAG ACC ACT CCC ATCCTT TCT-3’) and an antisense primer (5’-GAT GTG GCC ATC ACA TTCGTC AGA T-3’). The PCR product resulted in a 490 bp (I/I) and 190 bp (D/D) fragment analyzed on a 2% agarose gel stained with ethidium bromide。

ACE基因插入或缺失的聚合酶链反应（PCR）筛选用上游引物（5’-CTG GAG ACC ACT CCC ATCCTT TCT-3’）和反义引物（5’-GAT GTG GCC ATC ACA TTCGTC AGA T-3’）。PCR产物为490 bp（I / I）和190 bp（D / D）片段，并通过用溴化乙锭染色的2%琼脂糖凝胶进行分析。

The AT1R 1166A>C polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis with the following primers to amplify of the AT1R gene: forward, 5’-GCA GCA CTT CAC TAC CAA ATG AT-3’; and reverse, 5’-TGT TCT TCG AGC AGC CGT-3’.

AT1R 1166a＞C多态性是被聚合酶链反应限制性片段长度多态性（PCR-RFLP）分析下列引物扩增分析AT1R基因：向前，5’- GCA GCA CTT CAC TAC CAAATG AT-3”；反向，5’-TGT

有关基因的资料

TCT TCG AGC AGC CGT-3’.

The 176 bp fragment yielded by PCR amplification was incubated with 3U of BclI restriction enzyme for 16 h at 50℃. The 176 bp digestion fragment represents the normal homozygote (CC), the 176 bp and 156 bp fragments represent the heterozygote (AC), and the 156 bp fragment represents the mutant （突变的）homozygote (AA) genotype.

通过PCR扩增法得到的176 bp片段在50℃温度下培养3U BclI限制性酶 16 h。176 bp的消化片段表现正常纯合子（CC），176 bp和156 bp的片段代表杂合子（AC），并156 bp的片段代表突变纯合子基因型（AA）。

To determine the AGT M235 T genotype, PCR-RFLP analysis was also performed with a sense primer (5’ -CAG GGT GCT GTC CAC ACT GGA CCC C-3’ ) and an antisense primer (5’ -CCG TTT GTG CAG GGC CTG GCT CTC T-3’ ). The length of the amplified fragment was 165 bp. After amplification, PCR products were digested by a restriction endonuclease (3U of PsyI) for 16 h at 37 ℃. The PsyI-restricted products were as follows: 165 bp (MM), 165 and 141 bp (MT), and 141 bp (TT).

为了确定AGT M235 T基因型，使用上游引物进行（（5’ -CAG GGT GCT GTC CAC ACT GGA CCC C-3’）和反义引物（5’ -CCG TTT GTG CAG GGC CTG GCT CTC T-3’）进行了PCR-RFLP分析。扩增片段长度为165 bp。扩增后，PCR产物在37 ℃温度下用限制性内切酶（3U的psyi）消化16 小时。psyi限制的产品如下：165 bp（MM），165和141 bp（MT），和141 bp（TT）。

2.3. Statistical analysis

Differences in the ACE, AT1R, and AGT genotype frequencies between the cases and the normal controls were compared using a chi-square test. The odds ratio (OR) and 95% confidence interval (CI) were used as a measure of the strength of the association between genotype frequencies and RSA. Statistical significance was accepted at a p < 0.05 level. All statistical analyses were performed with GraphPad Prism 4.0 (GraphPad Software Inc., San Diego, CA, USA).

病例组和正常对照组的ACE，AT1R与AGT基因型频率之间的差异的比较使用卡方检验。比值比（OR）和95%置信区间（CI）是用来作为一个衡量基因型和RSA之间关联强度的指标。统计意义是公认的在P＜0.05水平。所有的统计分析是用GraphPad Prism 4.0 （graphpad 软件有限公司，San Diego, CA, USA）。

The MDR method has been proposed by Ritchie et al. [10] and Moore and William [11], and implemented by Hahn et al. [12] and Ritchie et al. [13]. The MDR method is described in detail previously [10–13]. Briefly, the MDR was comprised of two steps. First, the best combination of multi-factors was chosen. Second,the combinations of genotypes are classified into high- and low-risk groups [14].

在MDR方法是由Ritchie[10]和威廉摩尔[11]和哈恩[12]等人提出的，并被Ritchie[13]等人实施的。MDR方法以前被描述的非常细节[13]。简单的讲，MDR是又两个步骤组成。第一，多因素最优配合的选择。第二，基因型的组合被分为高和低风险两个组[ 14 ]。

有关基因的资料

Interaction analysis was performed in the open-source MDR software package (v.2.0) available from www. http://wendang.chazidian.com. Using MDR analysis, we constructed all possible combinations of one to three polymorphisms. The HAPSTAT program was used to estimate the haplotype frequencies for the polymorphisms selected by MDR analysis with strong synergistic effects. 相互作用分析是用open-source MDR software package (v.2.0)软件进行分析，并可从www. http://wendang.chazidian.com下载。使用MDR分析，构建了三个多态性的所有可能的组合。该HAPSTAT程序被用来估计，通过强大的协同效应被选取的，多态性的基因单倍型频率。

HAPSTAT allows testing of the haplotype effects and haplotype-environment interactions by maximizing the (observed-data) likelihood that properly accounts for phase uncertainty and study design. The current versions of the HAPSTAT software (v.3.0) are available from the

HAPSTAT允许测试单倍型效应及单倍型与环境的相互作用的影响，通过增大（观测数据）似然值，正确地考虑相位不确定性的研究与设计。HAPSTAT软件的当前版本（V3.0）可从下载。

3. Results

The three SNPs studied were in complete Hardy-Weinberg equilibrium. The distribution of ACE, AT1R, and AGT allele（等位基因） and genotype frequencies （基因型频率）among the controls and the RSA patients is shown in Table 1.

三个SNP位点进行了完整的哈迪-温伯格平衡。对照组和RSA病例组的ACE，AT1R和AGT等位基因的分布和基因型频率在表1中所示。

The genotype of each polymorphism in the subjects was in Hardy–Weinberg equilibrium. For the ACE I/D polymorphism of the 251 patients analyzed, 44 (17.5%) had the ACE DD genotype, 130 (51.8%) had the ACE ID genotype, and 77 (30.7%) had the ACE II genotype.

每个患者多态性的基因型是哈代–温伯格均衡额。对ACE I / D多态性的251名患者的分析中，44（17.5%）有ACE DD基因型，130（51.8%）有ACE ID基因型，和77（30.7%）有ACE II基因。