类赖氨酰氧化酶1、转化生长因子β1在子宫腺肌病中的表达及临床意义

目的:探讨类赖氨酰氧化酶1(LOXL1)、转化生长因子β1(TGFβ1)在子宫腺肌病(AM)中的作用及临床意义。方法:采用免疫组织化学法检测30例AM患者在位内膜和异位内膜组织及20例正常子宫内膜组织中LOXL1、TGF-β1的表达,并对其蛋白表达量进行相关性分析。结果:LOXL1在AM异位内膜组、AM在位内膜组及对照组中表达的平均光密度(MOD)值分别为:0.22±0.10、0.15±0.09、0.15±0.06。AM异位内膜组LOXL1的表达高于AM在位内膜组和对照组(P<0.05),而AM在位内膜组和对照组的LOXL1表达无统计学差异(P>0.05);TGFβ1在AM异位内膜组、AM在位内膜组及对照组的相对表达量分别为:0.26±0.02、0.16±0.03、0.15±0.02,AM异位内膜组TGFβ1的表达高于AM在位内膜组和对照组(P<0.05),AM在位内膜组和对照组间TGF-β1的表达无统计学差异(P>0.05)。AM异位内膜组LOXL1蛋白与TGFβ1蛋白的表达呈正相关(r=0.402,P<0.05)。AM在位内膜组LOXL1蛋白与TGFβ1蛋白的表达无相关关系(r=0.24,P>0.05)。LOXL1在子宫大小>孕12周及腺肌瘤>5 cm的AM异位内膜组表达强于子宫大小≤孕12周和腺肌瘤≤5 cm的AM异位内膜组(P<0.05),LOXL1的表达量与患者痛经与否、月经量差异亦无明显关系(P>0.05)。TGFβ1在AM异位内膜中表达与患者痛经、月经量、AM病灶大小的差异均无明显相关性(P>0.05)。结论:LOXL1、TGFβ1在子宫腺肌病的异常表达可能参与子宫腺肌病的发生和发展。     

肾损伤分子-1、β2-微球蛋白、内皮素-1和肌酐在妊娠期高血压疾病并发急性肾损伤的临床研究

目的:探讨血清肌酐(SCr)、内皮素-1(ET-1)、肾损伤分子-1(KIM-1)和β2-微球蛋白(β2-MG)的水平在妊娠期高血压疾病并发早期肾损伤的临床意义。方法:将妊娠期高血压疾病并发肾损伤的孕产妇40例设为肾病组,同期正常孕产妇40例设为对照组,分别在孕龄30周、32周、34周、36周检测孕妇血SCr、ET-1、尿KIM-1和β2-MG的水平。结果:1肾病组与对照组比较,患者的年龄、孕次和流产史均无统计学差异(P0.05),尿量、尿素氮(BUN)、SCr及尿蛋白定量均存在统计学差异(P0.001);2肾病组和对照组整体比较,SCr、ET-1、KIM-1和β2-MG的组间、时间点间的交互作用均有统计学差异(P0.05);3孕36周SCr、孕32周KIM-1及孕34周ET-1和β2-MG水平肾病组与对照组组间存在统计学差异(P0.05);4对照组组内不同孕周间SCr、ET-1、KIM-1和β2-MG水平无统计学差异(P0.05);肾病组组内与孕30周时比,KIM-1水平在孕32周显著升高(P0.05),在孕34周、36周进一步升高(P0.01);ET-1和β2-MG水平在孕34周显著升高(P0.05),而SCr在孕36周显著升高(P0.05)。结论:妊娠期高血压疾病孕妇产前监控中,联合检测SCr、ET-1、KIM-1和β2-MG能及时发现肾损伤的存在;妊娠期高血压疾病并发肾损伤时KIM-1、ET-1和β2-MG的改变早于SCr的改变。     

多囊卵巢综合征患者卵泡液LIF、IL-1β及性激素水平与IVF-ET结局关系的研究

目的:探讨多囊卵巢综合征(PCOS)患者卵泡液中白血病抑制因子(LIF)、白细胞介素-1β(IL-1β)及性激素水平与IVF-ET结局的关系。方法:用酶联免疫双抗夹心法和时间分辨免疫荧光法前瞻性研究了行IVF-ET的11例PCOS患者、14例对照组患者卵泡液中IL-1β、LIF及雌二醇(E2)和孕酮(P)的定量表达。结果:PCOS组卵泡液中LIF为21.1±11.1pg/mL,P为191.9×103nmol/L,明显低于对照组(33.5±11.8pg/mL,305.9×103nmol/L,P<0.05);而PCOS组卵泡液IL-Iβ为39.9±11.5pg/mL,E2浓度为3334.00nmol/L,明显高于对照组(28.3±10.6pg/mL,2138.1nmol/L),P<0.05。PCOS组胚胎种植率为8.8%,临床妊娠率为18.2%,明显低于对照组(16.7%,42.9%),P<0.05;PCOS组OHSS发生率为27.3%,明显高于对照组(7.1%,P<0.05)。LIF与E2在两组患者呈负相关(r=-0.442,P=0.027)、LIF与LH/FSH比值在PCOS组呈负相关(r=-0.682,P=0.021);IL-Iβ与E2在PCOS组呈正相关(r=0.612,P=0.045);LH/FSH比值与P在PCOS组呈负相关(r=-0.780,P=0.005);LIF与IL-Iβ水平两者间无明显相关性。结论:LIF可能是PCOS患者低种植率的关键因子;IL-Iβ可能是PCOS患者在控制性超排卵过程中易发生OHSS的一个致病因子;卵泡液IL-Iβ、LIF受卵巢激素调控。     

一氧化氮与大鼠离体黄体细胞凋亡的关系及白细胞介素-1β对其影响

目的 :探讨一氧化氮在黄体退化中的作用及细胞因子对其影响。方法 :以孕马血清促性腺激素 -人绒毛膜促性腺激素处理的雌性 Wistar大鼠为模型 ,分离制备了黄体细胞。采用 3'-末端标记 (TUNEL)法分析了 NO供体硝普钠 (SNP)、白细胞介素 -1 β(IL-1 β)对离体黄体细胞凋亡的作用。应用比色法及高效液相色谱仪分别检测了 IL-1 β对大鼠离体黄体细胞 NO合酶 (NOS)活性和 NO释放量的影响。结果 :SNP可以剂量方式诱导黄体细胞凋亡 ,1 μmol/ L SNP即可显著增加大鼠离体孵育 1 2 h的黄体细胞凋亡率 (为对照组的 1 33±1 8% ,P<0 .0 5)。IL-1 β(1 0 ng/ ml)可增强 SNP的作用。此外 ,IL-1 β可显著增加离体孵育1 2 h黄体细胞的总 NOS活性 (由 0 .2 0 1± 0 .0 51 U/ mg protein增高到 0 .382± 0 .1 0 6 U/mg protein,P<0 .0 1 ) ,促进 NO生成。结论 :NO-NOS系统可能通过诱导黄体细胞凋亡的途径参与黄体退化过程 ,IL-1 β可增强 NO-NOS系统的活性     

Tumor necrosis factor receptor 1 expression and early spontaneous abortion.

OBJECTIVES: To compare tumor necrosis factor receptor 1 (TNFR1) protein expression in women with unexplained early spontaneous abortion (UESA) and normal pregnancy. METHODS: In a prospective study, 62 women with UESA and 60 with normal pregnancy were studied. Decidual membrane TNFR1 was detected by flow cytometry and immunohistochemistry, and serum soluble TNFR1 were by ELISA. Statistical analyses of resulting data employed the student's t test, the Fisher's Exact, and the nonparametric Wilcoxon test RESULTS: The percentage of membrane tumor necrosis factor receptor 1 (mTNFR1) positive decidual cells was 16.42+/-7.1 for women with UESA and 12.47+/-5.3 for women with normal pregnancy (p<0.05). The number of mTNFR1 positive cells was more in decidual stromal and vessel endothelial cells in women with UESA, and serum soluble tumor necrosis factor receptor 1 (sTNFR1) concentration was significantly higher than in women with normal pregnancy (554.56+/-126.7 pg/ml vs. 175.3+/-52.4 pg/ml; p<0.001). CONCLUSION: The overexpression of TNFR1 may contribute to the development of ESA.     

TGF-β_1在PCOS卵巢间质纤维化形成中的作用

目的:探讨TGF-β1在PCOS大鼠卵巢间质纤维化、包膜硬化形成中的作用。方法:利用脱氢表雄酮(DHEA)皮下注射的方法建立PCOS病理模型大鼠20只,用微粒子酶免分析法测定血清性激素E2、T、LH、FSH、LH/FSH、空腹胰岛素(FINS)水平,及光镜下观察PCOS大鼠卵巢的病理结构,透射电镜观察细胞超微结构来验证模型。采用免疫组化法检测PCOS组(n=20)卵巢细胞因子TGF-β1的表达,并与20只正常大鼠相比照。结果:TGF-β1在PCOS组各阶段卵泡卵母细胞、颗粒细胞的表达强度与对照组相比,差异无统计学意义(P>0.05)。而在窦状卵泡膜细胞和卵巢间质细胞中的表达,PCOS组显著高于对照组(P<0.01,P<0.05)。结论:多囊卵巢中TGF-β1表达异常,可能是导致多囊卵巢间质纤维化、包膜增厚的原因之一,TGF-β1也参与PCOS卵泡发育、闭锁的调控。     

Production of interleukin-1-like molecules by human sperm cells

Objective: To characterize and localize interleukin (IL)-1 and IL-1β in human sperm cells.

Design: Prospective and comparative study.

Setting: Andrology clinic of a university hospital.

Patient(s): Two groups of normogonadotropic men: 17 fertile men (donors with proved fertility) and 8 oligoteratoasthenospermic infertile men.

Intervention(s): None.

Main Outcome Measure(s): Evaluation of IL-1 and IL-1β levels and expression in sperm cells by immunohistochemical staining, immunoassay, and Western blot analysis.

Result(s): Both types of IL-1–like molecules (IL-1 and IL-1β) were expressed in the tail, neck, and head of sperm cells of fertile men and patients with oligoteratoasthenospermia. Swim-up sperm cells from fertile men and patients with oligoteratoasthenospermia secreted similar levels of IL-1–like molecules. The levels of IL-1β–like molecules were higher than those of IL-1–like molecules in both groups. The expressed IL-1–like molecules were characterized by the presence a 60-kd protein for both IL-1–like and IL-1β–like molecules. In some samples of both fertile men and infertile men with oligoteratoasthenospermia, 17-kd, 33-kd, and 45-kd IL-1β–like molecules were detected. Impairment of sperm function, such as decreased sperm count and motility and/or impaired morphology, was not related to the capacity of sperm cells to produce IL-1–like molecules.

Conclusion(s): IL-1 molecules originating in sperm cells may play a role in the physiologic functions of sperm cells (autocrine effect) and/or in cell–cell interactions within the testis (paracrine effect).     

Prenatal diagnosis of familial recessive PIGN mutation associated with multiple anomalies: A case report

ObjectiveWe present a novel homozygous splice site mutation in the PIGN gene identified by whole exome sequencing and explored the genotype–phenotype correlation.Case reportA healthy 32-year-old woman underwent an ultrasound at 13 + 5 weeks of gestation. The ultrasound revealed multiple anomalies again including cystic hygroma, omphalocele and a ventricular septal defect. The pregnancy was subsequently terminated, and whole exome sequencing revealed a novel homozygous splice site mutation in the PIGN gene c.963 G > A (p.Gln321Gln). The same variant was also detected by pedigree-based Sanger sequencing in both parents as heterozygous, while they had normal karyotypes.ConclusionOur case report enhances the phenotype–genotype correlation associated with homozygous loss of function mutations in the PIGN gene.     

Up-regulation of p21-activated kinase 1 by in vitro treatment with interleukin 1-beta and its increased expression in ovarian endometriotic cysts

We evaluated whether the proinflammatory cytokines can regulate p21-activated kinase (Pak1) expression in endometrial cells as well as whether its expression is increased in endometriotic cysts. We found that interleukin-1β up-regulates Pak1 expression in endometrial stromal cells (ESC) and that the immunoreactivity of Pak1 is increased in the endometriotic cysts.     

Difference in expression of collagen type I and matrix metalloproteinase-1 in uterosacral ligaments of women with and without pelvic organ prolapse

Objectives

The objective of this study was to determine whether or not the angiotensin-converting enzyme insertion/deletion (ACE I/D), angiotensin II type 1 receptor (AT1R), and angiotensinogen (AGT) gene polymorphisms are associated with idiopathic recurrent spontaneous abortions (RSAs) in Korean women.

Study design

A total of 251 patients with unexplained consecutive pregnancy losses, and 126 healthy controls with at least one live birth and no history of pregnancy loss.

Result

The odds ratios (ORs) of the ACE ID (OR = 2.423; 95% confidence interval (CI) = 1.417–4.142; p = 0.001) and the ACE II (OR = 2.050; 95% CI = 1.143–3.675; p = 0.018) for the ACE DD genotype were significantly different between patients with idiopathic RSA and controls; however, there were no significant differences between patients and controls with respect to the AT1R 1166A>C and AGT M235 T polymorphisms. In a haplotype-based analysis of I-A (p = 0.010), D-A (p = 0.004), I-A-T (p = 0.033), D-A-T (p = 0.0005), and D-C-T (p = 0.013) polymorphism pairs with synergistic effects derived by the MDR method in patients and in controls showed significant results.

Conclusion

This study suggests that ACE, AT1R and AGT polymorphisms and haplotypes are a genetic determinant for the risk of idiopathic RSA in Korean women.     

子宫内膜异位症患者卵巢颗粒细胞SF-1基因启动子甲基化状态的研究

目的:探讨子宫内膜异位症(endometriosis,EMS)患者卵巢颗粒细胞中类固醇生成因子-1(steroidogenic factor-1,SF-1)表达下降的可能机制。方法:应用重亚硫酸盐测序法(bisulfite sequencing PCR,BSP)检测EMS患者和正常女性卵巢颗粒细胞SF-1基因启动子区5'Cp G位点甲基化水平,将测得的序列与原始序列比对,统计甲基化位点、数量并分析EMS组和正常组甲基化程度。结果:BSP片段涵盖SF-1基因转录起始位点附近的13个Cp G位点(从Cp G-84到Cp G+168),EMS组SF-1基因启动子区域的整体甲基化水平显著高于对照组(P0.05),逐个位点的分析显示EMS组Cp G+7、+18、+21、+42、+54、+60、+132以及+146位点相较于对照组呈现高甲基化水平(P0.05);而EMS组Cp G+77、+121和+141位点的高甲基化水平尤为显著(P0.001)。结论:EMS患者卵巢颗粒细胞SF-1基因启动子区5'Cp G位点甲基化水平显著高于对照组,可能是EMS患者卵巢颗粒细胞SF-1 m RNA表达降低的机制。     

受精方式、女方年龄、生育史和取卵数对体外异常受精的影响

目的:探讨人卵子体外受精后单原核(PN)和多原核胚胎形成的影响因素,为提高正常受精率探寻可行的方法。方法:回顾性分析1 004个IVF周期和250个ICSI周期,共计15 364个卵细胞资料,研究胚胎原核形成与体外受精方式、女方年龄、生育史、获卵数的关系。结果:①ICSI周期的1PN率明显高于IVF周期,而3PN和4PN形成率则明显低于IVF(P<0.05)。②当女方年龄>35岁时,1PN、3PN和4PN形成率均显著高于年龄≤35岁者(P<0.05);当女方年龄在28￣35岁时2PN(正常受精率)生成率最高(P<0.05),此年龄段的患者施行ICSI时3PN生成率也最高(P<0.05)。③女方无生育史的1PN形成率上明显高于有生育史患者(P<0.05)。④获取6￣10枚卵组,2PN形成率明显高于1￣5枚卵组和>20个卵组(P<0.05);而对于1PN、3PN、4PN形成率,获11￣15枚卵组明显高于其他获卵数组(P<0.05);而在获卵数>20枚时ICSI组2PN(正常受精率)生成率显著降低。结论:人体外受精正常与否受到受精方式、女方年龄、生育史和取卵数目等因素的影响,因此,在进行辅助生殖技术(ART)时应综合考虑这些因素,有助于提高正常受精率,降低异常受精率。     

水通道蛋白1在子宫内膜异位症患者血清和腹腔液中的表达及意义

目的:研究子宫内膜异位症(EMT)患者血清和腹腔液中水通道蛋白1(AQP1)及血管内皮生长因子(VEGF)表达水平的变化及其相关性,探讨其在子宫内膜异位症发病中的作用。方法:采用酶联免疫吸附方法(ELISA),测定36例EMT患者和22例对照组血清和腹腔液中AQP1和VEGF的含量,进行相关性分析。结果:EMT组血清和腹腔液中AQP1、VEGF水平均高于对照组,差异均有统计学意义(P<0.05)。AQP1、VEGF在EMT组血清和腹腔液的表达均有显著正相关性(r=0.776,P=0.000;r=0.771,P=0.000);AQP1、VEGF在对照组血清和腹腔液的表达则均无相关性(r=-0.026,P=0.910;r=-0.040,P=0.860)。结论:AQP1在EMT患者血清和腹腔液中过表达,可能推动异位内膜黏附和侵袭,AQP1与VEGF可能通过促血管生成,共同促进异位子宫内膜种植存活,是EMT形成的基础。     

雌、孕激素受体、生长激素受体、胰岛素样生长因子1受体与子宫内膜容受性的关系

辅助生殖技术(ART)经过近40年的发展,已取得突飞猛进的进展。但是临床妊娠率仍然徘徊在30%~40%。决定能否成功妊娠的因素主要包括2个方面,一是胚胎发育的质量,二是子宫内膜对胚胎的容受性。而在临床工作中,即使胚胎质量良好,种植率仍然不能令人满意。因此,如何提高子宫内膜容受性已经成为辅助生殖领域一个亟待解决的问题,了解影响子宫内膜容受性的相关因素,对于在种植窗期如何调节母体的内膜达到最佳的接纳胚胎的状态,具有重要的指导性意义。本文着重阐述子宫内膜容受性与雌激素受体(ER)、孕激素受体(PR)、胰岛素样生长因子1受体(IGF-1R)、生长激素受体(GHR)之间的关系,期望对临床判断子宫内膜容受性有所帮助。