胎儿颈项透明层厚度与不良妊娠结局

目的 探讨胎儿颈项透明层(nuchal translucency,NT)厚度增加与妊娠结局的关系,以期用NT预测胎儿临床预后. 方法 研究对象为2009年2月13日至2010年5月16日在昆明医科大学第一附属医院门诊就诊并行超声检查的妊娠早期单胎孕妇1839例.参照英国胎儿医学基金会的指南测量胎儿头臀径及胎儿NT厚度,随访妊娠结局.将预测不良妊娠结局发生高风险的界值分为NT≥2.0、2.5、3.0、3.5、4.5 mm,应用不同的界值,通过绘制受试者工作特征曲线评估NT增厚对不良妊娠结局的预测作用,观察NT增厚对临床结局的影响.用卡方检验或Fisher精确概率法分析敏感性和假阳性率之间的差异. 结果 1839例孕妇发生不良妊娠结局49例,不良妊娠结局发生率为2.7％.NT界值分别为2.0、2.5、3.0、3.5及4.5 mm时,预测总不良妊娠结局的阳性似然比分别为1.6、3.4、12.4、51.0及92.0,不同NT范围不良妊娠结局的发生率依次为2.7％、1.0％、4.0％、17.0％及75.0％.正常妊娠结局的比例随着NT增厚而降低.NT预测总不良妊娠结局的受试者工作特征曲线下面积为0.647,95％CI为0.561～0.732. 结论 妊娠早期胎儿NT增厚与多种不良妊娠结局相关.不同界值下NT预测不良妊娠结局的发生率可为临床咨询提供客观依据,且NT预测不良妊娠结局的效力达中等水平.     

薄型子宫内膜新鲜胚胎和冻融胚胎移植妊娠结局比较

目的探讨在薄型子宫内膜患者中新鲜胚胎移植与冻融胚胎移植(FET)妊娠结局的差异。方法回顾性分析接受体外受精/卵胞质内单精子显微注射-胚胎移植(IVF/ICSI-ET)治疗采用长方案胚胎移植h CG注射日与冻融周期胚胎移植内膜转化日的内膜厚度≤7 mm的患者共592个周期的临床资料。将移植周期按胚胎是否冻融分为新鲜胚胎移植组(n=173)和FET组(n=419)。比较组间的胚胎种植率、临床妊娠率、流产率、多胎率和异位妊娠率有无差异。结果新鲜胚胎移植组患者平均移植胚胎(2.1±0.4)枚,与FET组患者平均移植胚胎(2.1±0.5)枚比较,组间有统计学差异(P0.05);按照移植胚胎数分为3个亚组,新鲜胚胎移植组1枚胚胎者,妊娠率为7.7%,2枚者为30.2%,3枚者为23.8%;FET组1枚胚胎者15.6%,2枚者为34.9%,3枚者为41.6%,新鲜胚胎移植组与FET组间差异均无统计学意义(P0.05)。组间着床率、流产率、异位妊娠率等结果也均无统计学差异(P0.05)。移植3枚胚胎新鲜组多胎率(80.0%)高于FET组(29.7%)(P0.05)。新鲜胚胎移植组多胎率3个亚组间有统计学差异(P0.05),FET组妊娠率和流产率3个亚组间均有统计学差异(P0.05)。将移植胚胎数作为协变量,纳入Logistics回归模型对结果变量进行分析,说明周期类型与临床妊娠率间无显著相关性(OR=0.726,95%CI=0.504~1.104)。结论子宫内膜厚度≤7 mm的薄型内膜患者新鲜胚胎移植和FET妊娠结局相似,选择新鲜周期移植不影响妊娠结局并可缩短治疗周期,降低总费用。     

子宫内膜厚度对体外受精-胚胎移植治疗结局的影响

目的:探讨超促排卵(COH)周期中注射hCG当日的子宫内膜厚度对体外受精-胚胎移植治疗结局的影响。方法:回顾性分析502个IVF/ICSI周期,根据注射hCG当日子宫内膜厚度分为5组,A组:内膜厚度≤9mm,38个周期;B组:内膜厚度10-11mm,111个周期;C组:内膜厚度12-13mm,169个周期;D组:内膜厚度14-15mm,118个周期;E组,内膜厚度≥16mm,66个周期。结果:C组和D组的临床妊娠率(65.68%,67.80%)和种植率(42.59%,43.76%)显著高于A组(65.68%,33.23%)、B组(50.45%,34.31%)和E组(53.03%,31.89%)。结论:注射hCG当日子宫内膜的厚度影响了IVF-ET的治疗结局,厚度处于12-15mm时,更适宜着床。     

胚胎复苏后体外孵育时间对妊娠结局的影响

目的 探讨复苏后的体外孵育时间对冻存胚胎妊娠结局的影响。方法 对100例接受冷冻胚胎移植患者的临床资料进行回顾性分析。结果 100例冷冻胚胎移植周期，临床妊娠率为21％，复苏后体外孵育时间的不同妊娠率差异无显著性，但流产率差异有显著性。结论 复苏后延长体外孵育时间可以更好的观察胚胎的发育潜能，但增加流产率，应缩短复苏后体外孵育时间。     

移植8cell胚胎个数与妊娠率的关系

目的:探讨体外受精-胚胎移植(IVF-ET)周期中第三天移植8cell胚胎数量与临床结局的关系。方法:以2009年1月至2010年12月在本中心接受IVF-ET治疗625个周期为研究对象。将所有移植周期按移植8cell胚胎个数分为三组,分析三组的临床妊娠率、胚胎种植率、多胎率有无差异。结果:移植两个8细胞胚胎组(C组)的临床妊娠率(59.5%)显著高于无8细胞胚胎移植组(A组)和含一个8细胞组(B组)(P<0.05),A、B两组临床妊娠率无显著性差异;C组种植率(39.8%)显著高于其他两组,A组和移植胚胎中含一个8细胞组(B组)种植率比较有显著性差异,;多胎率分析,C组明显高于其他两组。结论:在IVF-ET周期中第3天移植8cell胚胎个数对妊娠率及种植率、多胎率有影响,在挑选胚胎时移植一个8cell胚胎搭配一个其他细胞数的胚胎也能得到较好的妊娠率并有效降低多胎率。     

移植8cell胚胎个数与妊娠率的关系

目的:探讨体外受精-胚胎移植(IVF-ET)周期中第三天移植8cell胚胎数量与临床结局的关系。方法:以2009年1月至2010年12月在本中心接受IVF-ET治疗625个周期为研究对象。将所有移植周期按移植8cell胚胎个数分为三组,分析三组的临床妊娠率、胚胎种植率、多胎率有无差异。结果:移植两个8细胞胚胎组(C组)的临床妊娠率(59.5%)显著高于无8细胞胚胎移植组(A组)和含一个8细胞组(B组)(P<0.05),A、B两组临床妊娠率无显著性差异;C组种植率(39.8%)显著高于其他两组,A组和移植胚胎中含一个8细胞组(B组)种植率比较有显著性差异,;多胎率分析,C组明显高于其他两组。结论:在IVF-ET周期中第3天移植8cell胚胎个数对妊娠率及种植率、多胎率有影响,在挑选胚胎时移植一个8cell胚胎搭配一个其他细胞数的胚胎也能得到较好的妊娠率并有效降低多胎率。     

胚胎冻融后体外培养时间对妊娠结局的影响

目的:探讨人卵裂期胚胎玻璃化冷冻-复苏后体外培养时间对妊娠结局的影响。方法:回顾性分析623例冻融胚胎移植(FET)周期的资料,根据解冻后不同移植时间分为当日移植组(A组,n=307)和次日移植组(B组,n=316);再根据卵裂球的生长与否将B组分为全部生长(B1组,n=184),部分生长(B2组,n=101)和均无生长(B3组,n=31)3组。比较各组间的临床妊娠率和胚胎种植率。结果:A组临床妊娠率和种植率分别为36.2%和19.6%,B组为38.3%和20.1%,组间差异无统计学意义(P>0.05);次日移植的B1组临床妊娠率和种植率(41.8%和23.4%)最高,但与B2组(39.6%和22.8%)比较差异无统计学意义(P>0.05);B3组最低(12.9%和5.4%),与B1、B2组比较有统计学差异(P<0.01)。结论:延长冻融胚胎体外培养时间不能显著提高冻融胚胎移植周期的妊娠率和种植率,延长培养时间后,无生长的胚胎发育潜能低。因此可以通过观察复苏后胚胎的生长情况评估其发育潜能,选择优质的胚胎进行移植。     

精子与卵母细胞透明带结合对ICSI治疗结局的影响

目的:探讨使用透明带结合后的精子行ICSI,是否可改善ICSI治疗结局。方法:选择上一周期IVF完全不受精者或男方为少、弱、畸形精子症而接受ICSI治疗的不育患者,共收录106对不孕夫妇,将其随机分成常规ICSI对照组和利用未成熟卵透明带上结合的精子进行ICSI的试验组,每组53例,分析比较ICSI结局。结果:试验组的卵裂率、优质胚胎率、可使用胚胎率均显著高于对照组,差异有统计学意义(P<0.05)。虽然临床妊娠率试验组(36%,19/53)与对照组(49%,26/53)组间无统计学差异(P>0.05),但试验组种植率(29%)、多胎率(35.71%,10/26)明显高于对照组(17%、10.53%,2/19)(P<0.01)。结论:通过利用患者自体ZP结合的精子进行ICSI注射,能获取高质量胚胎和高种植率,这种改良的ZP结合ICSI方法或许能为今后ICSI治疗提供一个新的思路。     

不同胚胎选择体系对IVF-ET妊娠结局的影响

目的:探讨IVF-ET周期中不同胚胎选择体系对妊娠结局的影响。方法:将530个IVF-ET周期随机分为3组,按不同的胚胎选择方法选择胚胎进行移植。A组:228个移植周期,根据d3胚胎常规评分方法选择胚胎;B组:140个移植周期,根据d3胚胎常规评分结合原核评分选择胚胎;C组:162个移植周期,根据d3胚胎常规评分结合胚胎早期分裂选择胚胎。将C组进一步分为移植胚胎中有早裂胚胎和无早裂胚胎二组。比较各组的临床妊娠率和单胚着床率。结果:B组的临床妊娠率和单胚种植率为37.86%和21.77%,与A组(33.33%和20.56%)比均无统计学差异(P>0.05);而C组(45.06%和30.88%)均显著高于A组(P<0.05);C组移植胚胎中有早裂胚胎者的临床妊娠率和单胚着床率为55.68%和38.38%,分别显著高于移植胚胎中不含早裂胚胎者(32.43%和21.94%)(P<0.05)。结论:d3胚胎常规评分结合胚胎早期分裂选择的胚胎移植,可以提高IVF-ET周期的临床妊娠率和单胚着床率。     

宫腔镜检查对反复胚胎种植失败者再次种植妊娠率的影响

目的:探讨宫腔镜检查对反复胚胎种植失败(RIF)患者再次胚胎种植的临床妊娠率(CPR)的影响。方法:回顾性分析RIF患者281例的临床资料,再次种植前接受宫腔镜检查为A组(210例),其中宫腔镜下无异常发现为A1组(144例),宫腔镜下有异常发现为A2组(66例),再次种植前未接受宫腔镜检查为B组(71例)。宫腔镜手术中对宫腔异常者予以处理,宫腔无异常者仅行内膜搔刮。3组患者均再次行胚胎种植,比较3组再次种植的CPR。结果:A1组CPR为25.7%,A2组CPR为30.3%,B组再次种植CPR为11.3%,A1组、A2组的CPR均高于B组(P0.05),A1组与A2组的CPR比较,差异无统计学意义(P0.05)。A2组中子宫内膜息肉(22例)、宫腔粘连(12例)患者的再次种植CPR(45.5%,41.7%)明显高于B组(11.3%),差异有统计学意义(P0.05);宫腔偏小(29例)患者的再次种植CPR(17.2%)与B组比较,差异无统计学意义(P0.05);宫腔镜下发现黏膜下子宫肌瘤、鞍状子宫、单角子宫各1例,宫腔镜术后再次胚胎种植均未妊娠。结论:宫腔镜下发现并处理宫腔异常,特别是子宫内膜息肉与宫腔粘连的治疗能提高再次种植的CPR;宫腔镜无异常发现,仅行内膜搔刮亦能提高再次种植的CPR。     

Improved implantation rate after chemical removal of the zona pellucida

OBJECTIVE: To evaluate the implantation rate achieved after chemical removal of the zona pellucida from day 5 human in vitro-derived embryos. DESIGN: Prospective, randomized, controlled study. SETTING: A tertiary care infertility clinic. PATIENT(S): Two hundred fifty-seven patients undergoing IVF with transfer of morulas or blastocysts on day 5 after oocyte retrieval. All patients had had at least two previous implantation failures. INTERVENTION(S): Chemical removal of zona pellucida by using acidic Tyrode's solution vs. no removal (controls). MAIN OUTCOME MEASURES: Clinical pregnancy rate and implantation rate per transfer. RESULT(S): Embryos without zona pellucida implanted at nearly twice the rate of control embryos (15.7% vs. 27.5%). The pregnancy rate was also significantly higher in the zona pellucida-free group than the control group (31.0% vs. 46.1%). Removal of zona pellucida was most effective in embryos with delayed development, which reached the morula or early cavitating stage on day 5 of in vitro culture (implantation rate, 12.1% vs. 25.7%). CONCLUSION(S): Chemical removal of zona pellucida from day 5 in vitro cultured human embryos is an effective and safe method of significantly improving the implantation rate, especially of embryos with delayed development.     

Influence of elevated pH levels on structural and functional characteristics of the human zona pellucida: Functional morphological aspects

Purpose A total of 86 fresh and salt-stored immature human oocytes derived from postmortem ovarian tissue were used for this study.Methods Oocytes were randomly incubated either in synthetic human tubal fluid medium (untreated zonae) or in a chemically defined medium (treated zonae).Results Sperm binding experiments using hemizona assay conditions exhibited a 10-fold increased binding of sperm to treated compared to untreated oocytes (272.7±43 versus 24.3±15 sperm bound, respectively; P<0.0001). pH recordings during incubation showed elevated pH levels of 8.1 compared to pH 7.2 among treated and untreated zonae, respectively. Ultrastructural examination showed a spongy appearance of the surface of treated zonae, whereas untreated zonae appeared compact with smooth surface.Conclusions The marked increase in sperm binding among treated zonae, together with the ultrastructural findings, suggest that the altered zona surface enhances sperm binding. The physiological maturational process of the zona pellucida might be manipulated in vitro, thus increasing sperm binding to the zona.Presented at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, April 3–7, 1995. Vienna, Austria.     

Chemical removal of the outside of the zona pellucida of day 3 human embryos has no impact on implantation rate

Two hundred eighteen consenting patients entered a randomized study of the application of chemical zona pellucida thinning on their day 3 embryos, prior to uterine transfer. Of those control patients (n =108), whose embryos remained unmanipulated, 40 (37.0%) have ongoing/delivered pregnancies, while in the experimental group (n =110), whose embryos had their zonae pellucidae chemically thinned, there are 49 patients (44.6%) who have ongoing/delivered pregnancies. Although this difference is not significant, clearly the application of this micromanipulative intervention has not been detrimental, and this bodes well for routine application of embryonic micromanipulation procedures in general. Certain patient subgroups were studied including older women, those with elevated basal follicle stimulating hormone levels, patients with embryos of differing zona thickness, and patients with embryos of differing uniformity of zona thickness. No significant influence of chemical removal of the outside of the zona on the implantation rate of embryos in any of these subgroups was observed other than a marginally significant (P =0.095) improvement of implantation of embryos with less than 4.0 µm variation in zona thickness when chemical zona thinning was applied. Failure of chemical zona thinning to enhance human embryo implantation significantly, compared to assisted hatching by complete zona drilling, strongly suggests that the bilayered human zona pellucida needs to be fully breached, unlike that of the mouse.     

Production of monoclonal antibodies to porcine zona pellucida and their inhibition of sperm penetration through human zona pellucida in vitro

Two hybridoma cell lines producing murine monoclonal antibodies to antigens common to the zona pellucida (ZP) of pigs and humans were obtained by immunization of mice with solubilized porcine zona antigen. Indirect immunofluorescence tests showed that both these monoclonal antibodies stained the entire layer of porcine ZP but stained different regions of human ZP, one staining the entire layer and the other only the outer surface. At high concentrations, these two monoclonal antibodies directed against antigens common to porcine and human ZP prevented sperm binding and penetration into human ZP in vitro, whereas a monoclonal antibody directed against an antigen restricted to porcine ZP did not have these inhibitory effects. It is concluded that human and porcine ZP share at least two antigens with different locations in the ZP, and that these influence or are essential for interaction of human sperm with the ZP. These results provide a rationale for using porcine ZP clinically as a vaccine for human immunocontraception.     

Enzymatic characterization of zona pellucida hardening in human eggs and embryos

Purpose To characterize possible hardening of the human zona pellucida (ZP) and evaluate the effect of culture duration, patient age, and ZP thickness, ZP of unfertilized eggs (experiment 1, n =367; experiment 2, n =174) and abnormal embryos (experiment 1, n=52) were randomly designated for -chymotrypsin treatment after 0, 24, 48, 72, 96, 120 h (experiment 1) and 48 h, 72 h, and 1 week (experiment 2) of in vitroculture in HTF medium supplemented with 0.5% human serum albumin. Mean ZP thickness was predetermined in experiment 2.Methods The dispersion of the ZP glycoproteins was assessed, and the duration of time for complete digestion was recorded as an index of ZP hardness.Results In experiment 1, enzyme digestion duration increased (P<0.05) in the first 24 h in vitrofrom 18.0±2.0 to 34.6±2.5 min, and tended to decrease over the next 4 days in culture (25.2±1.3, 29.4±0.9, 27.3±0.6, 26.6+1.1, and 20.7±1.5 min on Day 2– 6 ZP, respectively). Zona hardening of fertilized eggs was revealed by a longer (P<0.01) digestion time (32.2±1.8 vs 25.8±0.6 min).Conclusions There were significant patient-to-patient variations (16.4±0.7 to 39.6±2.2 min); however, age was not correlated to enzyme digestion duration. In experiment 2 we determined that ZP thickness (range 8.4–21.6 m; mean 14.6±0.2 m) was not correlated (r=0.09) to the digestion interval (mean 24.3 ± 0.8 min). Based on our enzymatic ZP digestion measurements, it is apparent that spontaneous zona hardening does occur within 24 h of in vitroculture, similar to levels achieved postfertilization. The data do not support, however, the concept that additional, abnormal hardening of the ZP occurs during extended culturing.Presented at the 50th Annual Meeting of the American Fertility Society, San Antonio, Texas, November 5–10, 1994.     

Hemizona assay: Use of fresh versus salt-stored human oocytes to evaluate sperm binding potential to the zona pellucida

Salt-stored human oocytes (pH 7.0) showed sperm binding ability equal to that of fresh, living oocytes under hemizona assay (HZA) conditions.     

Apoptosis in human cumulus cells in relation to zona pellucida thickness variation, maturation stage, and cleavage of the corresponding oocyte after intracytoplasmic sperm injection

OBJECTIVE: To assess the degree of apoptosis in the cumulus cells and the variation of the zona pellucida and the maturity and fertilization of the corresponding oocyte. DESIGN: Retrospective study. SETTING: Private fertility clinic. PATIENT(S): Fifty couples undergoing ICSI. INTERVENTION(S): ICSI. MAIN OUTCOME MEASURE(S): Correlation between apoptosis in the cumulus cells and the zona pellucida thickness variation, maturation stage, fertilization rate, and embryo score. RESULT(S): This study demonstrated no correlation between apoptosis in cumulus cells and the thickness and variation of the zona pellucida in oocytes and embryos. The incidence of apoptosis was significantly higher in cumulus cells from empty zona pellucidas and germinal vesicle stage and metaphase I oocytes compared with metaphase II oocytes. Non-fertilized metaphase II oocytes showed significantly higher incidence of apoptosis compared with fertilized metaphase II oocytes. There was a correlation between embryo score and the zona pellucida thickness variation. CONCLUSION(S): Apoptosis in cumulus cells had no impact on the zona pellucida thickness and variation in oocytes and embryos. The zona pellucida thickness variation was positively correlated to good embryo score. A higher degree of apoptosis was seen in cumulus cells from immature oocytes compared with mature oocytes. Furthermore, apoptosis in cumulus cells impaired the fertilization rate of metaphase II oocytes after ICSI.     

Infertility and Immunocontraception based on zona pellucida

The zona pellucida (ZP) is an extracellular matrix surrounding ovarian oocytes, ovulated eggs and preimplantation embryos. It plays several important roles at different stages of reproduction. Its constituent glycoproteins are expressed specifically in the ovary. It is thus possible to produce autoantibodies to ZP proteins that interfere with reproductive functions including folliculogenesis, fertilization and implantation. First, this article describes the history of anti-ZP antibodies detected in women with idiopathic infertility. Second, the current relationship between anti-ZP antibodies and infertility is discussed in relation to assisted reproductive medicine. Third, we introduce the latest studies of animal experiments involving the ZP. Finally, immunocontraceptive vaccine development using various ZP antigens is reviewed.     

Effects of aging on the zona pellucida surface of mouse oocytes

In a scanning microscopy study of mouse oocytes from immature, young, and aged females and of oocytes agedin vivo orin vitro, we have observed four types of zona pellucida, which we classify as types A, B, C, and D. Oocyte aging gives rise to a significant increase in predegenerative (type C) oocytes; this type of zona pellucida surface could result from a zona hardening effect and decrease the rate of fertilization of the oocytes affected.     

A comparison of the effects of different degrees of zona pellucida damage followed by cryopreservation on the postthaw development of mouse embryos

Purpose: The totally intact zona pellucida is not essential for the development of embryos. It is still unclear how much effect the degree of damages to the zona pellucida will have on the developmental potential of postthaw embryos after cryopreservation. We compared the developmental potential of cryopreserved mouse embryos after induction of two degrees of mechanical damage to the zonae pellucidae by micromanipulation. Methods: In experiment I, the development of 124 cryopreserved ICR mouse embryos to the blastocyst stage after zona pellucida penetration of two-cell embryos as in the procedures of subzonal sperm insertion (SUZI) was compared with the development of zona-intact cryopreserved embryos. In experiment II, the zonae pellucidae of 93 two-cell mouse embryos were dissected as in the procedures of partial zonal dissection (PZD), following which the embryos were frozen. This postthaw development was also compared with that of zona-intact two-cell cryopreserved embryos. All the embryos were thawed and cultured to the blastocyst stage. Additional controls were provided by culturing zonaintact and zona-penetrated or zona-dissected embryos without cryopreservation. Results: The development of unfrozen mouse embryos was not affected by either zona penetration (P=0.433) or zona dissection (P=0.659). The developmental potential of cryopreserved mouse embryos was significantly affected after zona dissection (blastocyst rate, 31% ZD vs 72%, control; P<0.001) but not after zona penetration (blastocyst rate, 59% ZP vs 64% control; P=0.441). Conclusions: The quality of cryopreserved embryos was affected by a large hole on the zona pellucida created by zona dissection but not by simple zona penetration.