Associations between toxic metals in follicular fluid and in vitro fertilization (IVF) outcomes

Purpose

We previously reported associations between trace concentrations of Hg, Cd and Pb in blood and urine and reproductive outcomes for women undergoing in-vitro fertilization (IVF). Here we assess measurements in single follicular fluid (FF) specimens from 46 women as a presumably more relevant marker of dose for reproductive toxicity.

Methods

FF specimens were analyzed for Hg, Cd and Pb using sector field-inductively coupled plasma-mass spectrometry (SF-ICP-MS). Variability sources were assessed by nested ANOVA. Multivariable regression was used to evaluate associations for square root transformed metals with IVF outcomes, adjusting for confounders.

Results

An inverse association is detected for FF Pb and fertilization (relative risk (RR) = 0.68, P = 0.026), although positive for Cd (RR = 9.05, P = 0.025). While no other statistically significant associations are detected, odds ratios (OR) are increased for embryo cleavage with Hg (OR = 3.83, P = 0.264) and Cd (OR = 3.18, P = 0.644), and for embryo fragmentation with Cd (OR = 4.08, P = 0.586) and Pb (OR = 2.22, P = 0.220). Positive estimates are observed for Cd with biochemical (RR = 19.02, P = 0.286) and clinical pregnancies (RR = 38.80, P = 0.212), yet with very low precision.

Conclusions

We have identified associations between trace amounts of Pb and Cd in FF from a single follicle, and oocyte fertilization. Yet, the likelihood of biological variation in trace element concentrations within and between follicles, coupled with levels that are near the limits of detection suggest that future work should examine multiple follicles using a ‘one follicle-one oocyte/embryo’ approach. A larger study is merited to assess more definitively the role that these environmental factors could play with respect to egg quality in IVF programs.     

脂质蓄积指数较腰围更好反映多囊卵巢综合征患者的胰岛素抵抗

目的:比较多囊卵巢综合征(PCOS)人群脂质蓄积指数(LAP)和腰围(WC)与胰岛素抵抗(IR)的相关性。方法:纳入121例PCOS患者和115例正常对照,测量人体参数及血液代谢指标,行75 g口服葡萄糖耐量试验,对其中20例对照及90例PCOS患者行高胰岛素-正葡萄糖钳夹术,并计算HOMA2-IR及LAP。结果:PCOS组LAP、WC等较对照组明显增高(P<0.05),胰岛素敏感性明显降低(P<0.05);匹配WC和年龄后,PCOS组LAP仍明显高于对照组(P<0.01),胰岛素敏感性明显低于对照组(P<0.05);与WC相比,LAP与M值(r=-0.66 vs r=-0.68,P<0.01)及HOMA2-IR(r=0.49 vs r=0.55,P<0.01)间关系更密切,校正WC后,LAP仍与M值(r=-0.37,P<0.01)及HOMA2-IR(r=0.26,P<0.01)相关;二项Logistic回归显示,高LAP患者发生IR的风险是正常LAP患者的6.05倍(OR=6.05,95%CI:1.67,21.90,P<0.05)。结论:在PCOS人群中,LAP与胰岛素敏感性相关,较WC更好地反映IR。     

长效孕激素炔诺酮庚酸酯及复方18甲基炔诺酮对血清性激素结合球蛋白的影响

24例妇女肌注炔诺酮庚酸酯200 mg,肌注前血清性激素结合球蛋白(SHBG)为53.32±23.23 nmol╱L。注射后5天为41.99±17.06nmol/L(P<0.05),21天降至最低值14.72±10.01 nmol/L,以后逐渐回升,直至注射后84天仍未回复到原来水平(34.46±20.39 nmol/L)。10例妇女单次口服18甲基炔诺酮6 mg+炔雌醚3 mg,服药前血清SHBG水平为42.76±13.89 n mol/L,服药后7天为62.53±10.90 nmol/L(P<0.01),14天达峰值为71.33±5.77 nmol/L,以后持续此高水平,直至服药后56天;8例连续3次口服18甲+炔雌醚,每次间隔23天,服药前血清SHBG水平为44.94±15.36 nmol/L,第1次服药后9天为83.46±10.08 nmol/L(P<0.01),16天达峰值为91.74±2.28nmol/L,以后持续高水平直至第3次服药后30天,第2、3次重复给药对业已升高的SHBG水平无影明显响。第2、3次服药后LNG峰值明显高于首次服药后峰值6倍。     

Maternal infibulation and obstetrical outcome in Djibouti

Objective: The objective of the present study was to assess the relation between female genital mutilation and obstetric outcome in an East African urban clinic with a standardized care, taking into account medical and socioeconomic status.

Methods: This was a cohort study conducted in Djibouti between October 1, 2012 and April 30, 2014. Overall 643 mothers were interviewed and clinically assessed for the presence of female genital mutilation. The prevalence of obstetric complications by infibulation status was included in a multivariate stepwise regression model.

Results: Overall, 29 of 643 women did not have any form of mutilation (4.5%), as opposed to 238 of 643 women with infibulation (37.0%), 369 with type 2 (57.4%), and 7 with type 1 mutilation (1.1%).Women with a severe type of mutilation were more likely to have socio-economic and medical risk factors. After adjustment, the only outcome that was significantly related with infibulation was the presence of meconium-stained amniotic fluid with an odds ratio of 1.58 (1.10–2.27), p value=0.014.

Conclusions: Infibulation was not related with excess perinatal morbidity in this setting with a very high prevalence of female genital mutilation, but future research should concentrate on the relation between infibulation and meconium.     

高雄激素血症与多囊卵巢综合征中非酒精性脂肪性肝病的关系

目的:探讨高雄激素血症(HT)与多囊卵巢综合征(PCOS)患者非酒精性脂肪性肝病(NAFLD)发病的关系。方法:对306例PCOS患者进行基础内分泌、口服糖耐量试验及胰岛素释放试验、肝功能、血脂等检查,B超诊断脂肪肝。结果:306例PCOS患者中诊断出NAFLD94例,发病率为30.7%;其中NAFLD轻度56例(59.6%),中度34例(36.2%),重度4例(4.2%)。PCOS患者NAFLD的发病率与HA无明显相关性,与稳态模型评估胰岛素抵抗指数(HOMA-IR)、甘油三酯(TG)呈正相关。PCOS合并NAFLD组胰岛素抵抗(IR)发生率明显高于PCOS不合并NAFLD组,P<0.05;PCOS高雄激素血症(PCOS-HA)组IR发生率与PCOS非高雄激素血症(PCOS-NHA)组无明显差异,P>0.05。结论:PCOS患者NAFLD发病率较高,PCOS患者NAFLD的发生与IR、代谢异常密切相关,与HA无明显相关。     

取卵日血清脑源性神经营养因子与辅助生育结局的相关性

目的:探讨取卵日血中脑源性神经营养因子(BDNF)能否预测卵母细胞质量和其后的卵胞质内单精子注射(ICSI)的妊娠结局。方法:总共纳入57例进行ICSI患者,在取卵日清晨抽取空腹静脉血,测量BDNF,E2和P的浓度。比较妊娠组与非妊娠组的相关数据,并分析取卵日BDNF水平与E2、P、年龄和IVF相关数据之间的相关性。结果:妊娠组与非妊娠组的BNDF浓度无明显差异;取卵日BDNF浓度与E2和P呈明显正相关。BDNF与获卵数和成熟卵母细胞数成正相关。结论:尚不能用取卵日BDNF来预测ICSI的妊娠结局,但BDNF可能在卵母细胞发育成熟过程中发挥着重要作用。     

非编码RNA在卵泡发育和卵巢疾病中的研究进展

非编码RNA(non-coding RNA,nc RNA)是目前生物医学领域的研究热点,如长链非编码RNA(long non-coding RNA,lnc RNA)和小非编码RNA:微小RNA(mirco RNA,mi RNA)、内源性小干扰RNA(endogenous small interfering RNA,endo-si RNA)、PIWI相互作用RNA(PIWI interacting RNA,pi RNA)等,它们作为细胞内基因调控网络的重要成员影响细胞的各种生命活动。近年来研究表明这些非编码RNA在卵巢也发挥重要作用,参与了颗粒细胞增殖、分化、凋亡以及激素合成的调控,在卵母细胞存活和发育中发挥重要作用,非编码RNA的表达异常与多囊卵巢等疾病密切相关,本文就非编码RNA与卵巢卵泡发育以及卵巢相关疾病的研究作一简述。     

多囊卵巢综合征患者脂联素、C反应蛋白水平的变化及其意义

目的:探讨脂联素(APN)、C反应蛋白(CRP)的变化对多囊卵巢综合征(PCOS)发病的意义。方法:按体重指数(BMI≥25kg/m2或<25kg/m2)分别将PCOS患者52例、对照组47例分为PCOS肥胖组(25例)、非肥胖组(27例)和对照肥胖组(23例)、对照非肥胖组(24例)4组。用ELISA法测定4组的APN水平、散射比浊法测CRP水平,葡萄糖氧化酶法测定空腹血糖(FPG)、化学发光法测空腹胰岛素(FIN)水平,并计算胰岛素抵抗指数(HOMA-IR)。结果:①PCOS组APN水平低于对照组(P<0.05),且同组肥胖者低于非肥胖者。②PCOS组CRP水平高于对照组(P<0.05),且肥胖者高于非肥胖者。③APN水平与BMI、HOMA-IR水平呈明显负相关(P<0.05)。结论:PCOS组APN水平降低,CRP水平升高,且以肥胖者明显。APN水平降低、CRP水平升高与PCOS患者胰岛素抵抗密切相关。     

细菌脂多糖对小鼠生长发育和骨骼发育的影响

目的:研究细菌脂多糖(LPS)对小鼠宫内胎儿死亡(IUFD)、生长发育迟缓(IUGR)和骨骼发育迟缓的影响。方法:LPS低中高组小鼠于妊娠d15-17分别腹腔注射不同剂量LPS(25μg/kg、50μg/kg、75μg/kg),LPS+2-苯叔丁基硝酮(PBN,活性氧ROS拮抗剂)组在LPS(75μg/kg)处理前30min和后3h经腹腔各给予100mg/kg的PBN,对照组给予等容量生理盐水。孕鼠于妊娠d18处死。另给药1d时取LPS高剂量组LPS+PBN组和对照组于LPS处理后6h处死孕鼠。结果:①小鼠妊娠d15-17给予LPS后,中高剂量组平均每窝死胎数明显高于对照组,活胎体重、身长和尾长下降,并呈明显的剂量-效应关系;LPS高剂量导致IUGR和骨骼发育迟缓;PBN处理明显抑制LPS对胎儿的作用。②LPS使母肝、胎肝和胎盘组织脂质过氧化,GSH含量显著降低。PBN显著抑制LPS的这些作用。结论:母鼠妊娠晚期接触LPS引起IUFD、IUGR和骨骼发育迟缓,ROS至少部分参与了LPS的引起IUFD、IUGR和骨骼发育迟缓。     

Ameliorative effect of myo-inositol on red blood cell alterations in polycystic ovary syndrome: in vitro study

Polycystic ovary syndrome (PCOS)is a gynecological endocrine disorder which is associated with systemic inflammatory status inducing red blood cells (RBC) membrane alterations related to insulin resistance and testosterone levels which could be greatly improved by myo-inositol (MYO) uptake. In this study we aim to evaluate the effect of MYO in reducing oxidative-related alterations through in vitro study on PCOS RBC. Blood samples from two groups of volunteers, control group (CG, n?=?12) and PCOS patient group (PG, n?=?12), were analyzed for band 3 tyrosine phosphorylation (Tyr-P), high molecular weight aggregate (HMWA), IgG in RBC membranes, and glutathione (GSH) in cytosol, following O/N incubation in the presence or absence of MYO. PCOS RBC underwent oxidative stress as indicated by higher band 3 Tyr-P and HMWA and increased membrane bound autologous IgG. Twenty four hours (but not shorter time) MYO incubation, significantly improved both Tyr-P level and HMWA formation and concomitant membrane IgG binding. However, no relevant modification of GSH content was detected. PCOS RBC membranes are characterized by increased oxidized level and enhanced sensitivity to oxidative injuries leading to potential premature RBC removal. MYO treatment is effective in reducing oxidative related abnormalities in PCOS patients probably restoring the inositol phospholipid pools of the membranes.     

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。     

Dehydroepiandrosterone sulfate/free androgen index ratio predicts a favorable metabolic profile in patients with polycystic ovary syndrome

Potential effect of hyperandrogenemia on metabolic disturbances in polycystic ovary syndrome (PCOS) has always been a matter of interest. We analyzed the records of 125 patients with PCOS and 54 age-matched healthy women. All participants underwent biochemical and hormonal assessment and a 75?g oral glucose tolerance test was performed. PCOS and control groups were comparable in terms of age. Dehydroepiandrosterone sulfate/free androgen index (DHEAS/FAI) ratio was negatively correlated with body mass index (BMI) (p?<?.001), fasting glucose (p?=?.02), area under the curve (AUC) of glucose (p?=?.03), AUC of insulin (p?=?.001), homeostasis model assessment-estimated insulin resistance (HOMA-IR) (p?<?.001), and triglycerides (TG) (p?=?.009), and positively correlated with insulin sensitivity index (ISI) (p?<?.001) and high-density lipoprotein cholesterol (HDL-C) (p?<?.001) among PCOS patients. In logistic regression analysis, higher DHEAS/FAI ratio levels were associated with lower risk of low HDL-C [RR(95%CI); 0.97(0.95–0.98); p?<?.001] as well as atherogenic dyslipidemia (TG/HDL-C) [RR(95%CI); 0.97(0.94–0.99); p?=?.035] even after adjustment for BMI in the PCOS group. Androgens, DHEAS and FAI act differently on metabolic parameters. Our results demonstrate that high DHEA-S/FAI ratio levels are associated with a more favorable metabolic profile.     

小鼠胚胎干细胞体外诱导分化为精原干细胞的研究

目的:初步探讨在体外条件下诱导小鼠胚胎干细胞(EGFP-mESC)分化为精原干细胞(SSC)的条件,建立有效的技术平台。方法:采用部分模拟胚胎早期发育的方法,将EGFP-mESC(XY)经体外悬滴培养,制备拟胚体(EB);使用免疫磁珠分选法从EB中分离出表达原始生殖细胞(PGC)表面标志SSEA-1的细胞;获得的细胞经2μmol/L维甲酸(RA)诱导增殖,使之向雄性生殖细胞分化。对诱导的细胞采用RT-PCR及免疫荧光检测特异性基因及蛋白的碱性磷酸酶。结果:在诱导EGFP-mESC向雄性生殖细胞分化的过程中,采用RT-PCR方法检测到了雄性生殖细胞特异表达基因Sry、fragilis、stella、Tex14等mRNA的表达;利用激光共聚焦方法检测到SSC表面标志蛋白Stra8、integrin-α6及Hsp90α的表达。结论:采用RA体外诱导EGFP-mESC定向分化为SSC是可行的。     

使用国产长效避孕皮下埋植剂妇女五年内血清雌二醇、孕酮、左旋18甲基炔诺酮水平

５例使用国产长效避孕皮埋剂（Ｓｉｎｏ－Ｉｍｐｌａｎｔ，以下简称为Ｓｉｎｏ）长达五年以上的妇女，每年末抽血一个周期（即连续４～５周，每周定时抽血一次）。用ＲＩＡ测定血清中雌二醇（Ｅ２），孕酮（Ｐ）和左旋１８甲基炔诺酮（ＬＮＧ）水平。研究结果显示：长期使用Ｓｉｎｏ未出现相似的卵巢内分泌反应。８４％（２１／２５）抽血周期血清Ｅ２水平出现峰值（Ｅ２＞１５０ｐｇ／ｍｌ），以例数计，埋植后１～４年内每年有８０％（４／５）抽血周期血清中Ｅ２显示峰值，第五年为１００％（５／５）。４０％（１０／２５）抽血周期显示黄体活性（Ｐ＞３ｎｇ／ｍｌ），以例数计，埋植第一年为０％（０／５）；第二年为４０％（２／５）；３～５年每年６０％（３／５）表现黄体活性。５例对象中有２例埋植后五年内未见血清Ｐ＞１ｎｇ／ｍｌ，而且这２例分别在埋植后１～２年及３～４年未见Ｅ２峰（Ｅ２＜１５０ｐｇ／ｍｌ）。５例对象埋植前对照周期均有正常的Ｅ２峰和黄体活性。在用Ｓｉｎｏ的５年内抽血周期血清Ｅ２和Ｐ的均值都低于对照周期。在用Ｓｉｎｏ期间，有黄体活性的周期均伴有Ｅ２峰。然而，并非所有有Ｅ２峰的周期都表现黄体活性。在埋植第一年，除第一个月外，血清ＬＮＧ水平低而相对?     

Changes in glandular and stromal estrogen and progesterone receptor isoform expression in eutopic and ectopic endometrium following treatment with the levonorgestrel-releasing intrauterine system

Objectives

The levonorgestrel (LNG) intrauterine system (LNG-IUS) has been shown to improve symptoms in women with minimal to moderate endometriosis. The precise mechanism for this is unknown. We hypothesized that this involves alteration in the expression of estrogen receptors (ER) and progesterone receptors (PR).

Study design

A prospective study of tissues obtained prospectively from 28 women with laparoscopically confirmed minimal to moderate endometriosis treated with LNG-IUS for 6 months. Endometrial and endometriotic biopsies obtained before and 6 months after treatment were processed and stained for ER-α, ER-β and PR expression by immunohistochemistry. Photographs were obtained and the receptors quantified.

Results

The mean (±SD) age of the 28 women was 31 ± 7.2 (range 18-42) years. Eight of them at initial biopsy were in the proliferative phase and 20 in the secretory phase. ER-α, ER-β and PR expression decreased significantly in the glandular (P < 0.0001) and stromal (P < 0.0001) compartments of the eutopic endometrium after treatment with LNG-IUS. Similarly, ER-α, ER-β and PR were significantly decreased in the stromal compartment of ectopic endometrium (P < 0.0001), and significantly decreased in the ectopic glands of ER-α (P < 0.0001), ER-β (P = 0.0002) and PR (P = 0.0064) expression.

Conclusion

The ameliorative effect of LNG-IUS on the symptoms of minimal to moderate endometriosis is likely modulated through a decrease in the expression of glandular and stromal ER-α, ER-β and PR in the ectopic endometrium.     

卵泡液中双酚A在多囊卵巢综合征发病中的作用及相关因素探讨

目的:探讨卵泡液中双酚A(bisphenol A,BPA)在多囊卵巢综合征(PCOS)发病中的作用。方法:于取卵日收集行体外受精/卵胞质内单精子显微注射(IVF/ICSI)的PCOS患者和非PCOS患者的卵泡液,检测患者卵泡液BPA水平和卵泡液性激素水平,并进一步与患者血清抗苗勒氏管激素(AMH)及临床结局等因素行相关性分析。结果:PCOS患者和非PCOS患者的卵泡液BPA水平(446.57±63.57 ng/L vs 336.29±59.02 ng/L,P0.001)、血清AMH水平(12.09±4.78μg/L vs5.69±2.98μg/L,P0.001)、卵泡液FSH(5.58±1.66 IU/L vs 3.95±1.54 IU/L,P=0.008)、LH(4.54±2.97 IU/L vs 0.96±1.10 IU/L,P0.001)和T(54.68±34.34 nmol/L vs 32.48±16.68 nmol/L,P=0.035)组间比较有统计学差异。相关性分析显示,卵泡液中BPA水平与血清AMH水平呈显著正相关(r=0.577,P=0.001)。结论:卵巢局部的BPA暴露可能参与了PCOS疾病的发生、发展。     

肥胖型多囊卵巢综合征并发妊娠期高血压/子痫前期风险因素的研究

目的探讨肥胖型多囊卵巢综合征(PCOS)患者并发妊娠期高血压/子痫前期(PIH/PE)的风险因素。方法对PCOS合并不孕患者通过促排卵成功单胎妊娠的妇女进行前瞻性队列研究,随访至产后12周。从孕前的临床体征评分、内分泌及代谢状态中筛选出PCOS并发PIH/PE的风险因素。结果本研究共纳入促排卵受孕的PCOS患者92例,其中肥胖型和非肥胖型分别为38例和54例,并发PIH/PE分别为11例和6例,发病率分别为28.9%和11.1%(P0.05)。Logistic回归多因素分析显示肥胖型PCOS患者并发PIH/PE的风险因素为血性激素结合球蛋白(SHBG)(OR=0.975,P0.05),是唯一的独立风险因子;受试者工作特征(ROC)曲线分析孕前SHBG和腰臀比(WHR)对肥胖型PCOS患PIH/PE的预测价值分别为0.883及0.775(P均0.05)。结论肥胖型比非肥胖型PCOS患者更易并发PIH/PE,肥胖型PCOS患者孕前的低SHBG、中心性肥胖都可能与孕期并发PIH/PE有关。