Lipofuscin bodies in human oocytes as an indicator of oocyte quality

Refractile bodies are one of the main morphological abnormalities that can be observed in the cytoplasm of human oocytes. In the present studies the characteristics of refractile bodies and the relationship between the size of these structures and developmental competence of the affected oocytes and resulting embryos were examined. The refractile bodies were found to have yellow autofluorescence which was consistent with the typical autofluorescence of lipofuscin. Viewed by transmitted electron microscopy, the refractile bodies showed the conventional morphology of lipofuscin inclusions and consisted of a mixture of lipids and dense granule materials. Large refractile bodies (>5 μm) were positively stained by the Schmorl reaction and were considered to contain lipofuscin. These larger lipofuscin inclusions (>5 μm) were associated with significantly reduced fertilization and unfavorable blastocyst development. Refractile bodies in human oocytes were found to contain lipofuscin. The largest lipofuscin inclusions (>5 μm) were associated with significantly reduced fertilization and unfavorable blastocyst development.     

低剂量17β雌二醇在赠卵助孕术中的应用

目的 :研究恒定、低剂量微粒化 1 7β雌二醇 (诺坤复 )在赠卵助孕术中的应用。方法 :1 999年 1月至 2 0 0 0年 7月 ,应用诺坤复 1～ 2 mg/ d和黄体酮为 43例接受赠卵的患者准备子宫内膜共 5 1个周期 ,由 40例同期行体外受精和胚胎移植 (IVF-ET)或配子输卵管内移植术 (GIFT)的不孕患者和 5名正常妇女提供卵母细胞。结果 :供卵和接受赠卵者共移植 90个周期 ,妊娠 34例 ,妊娠率为 38%。接受赠卵的 5 1个周期中 ,临床妊娠 2 0例 ,供卵者移植 39例中 ,临床妊娠 1 4例 ,妊娠率分别为 39%和 36 % ,两组间无显著差别 ,妊娠与未妊娠患者之间平均年龄、不孕原因和移植的胚胎数等均无显著差别。结论 :在等待赠卵期间 ,用低剂量 (1～ 2 mg/ d)诺坤复可有效地刺激子宫内膜生长 ,为胚胎移植作准备。     

樟脑丸对雌性小鼠生殖细胞和脏器的影响

目的 :探索由对二氯化苯制成的樟脑的生殖毒性。方法 :采用卵母细胞体外培养、体外受精等方法研究樟脑丸对小鼠卵母细胞成熟和受精能力的影响。结果 :接触樟脑丸 3 d组和 5d组对小鼠的卵巢和肝脏有明显的毒性作用 ,5d组能显著降低超排卵数 ;樟脑丸对卵母细胞生发泡破裂没有影响 ,但抑制卵母细胞第一极体的释放 ,影响其存活率及降低体外受精率。结论 :本实验结果提示 ,樟脑丸可能影响超排卵小鼠卵母细胞的成熟 ,明显降低卵母细胞的体外受精的能力 ,并表现有明显的生殖细胞毒性。     

镍对小鼠卵母细胞成熟和体外受精的影响

为了研究镍的生殖毒性 ,采用卵母细胞体外培养、体外受精的方法研究镍对小鼠卵母细胞的成熟和体外受精率的影响。结果 :氯化镍对卵母细胞生发泡破裂没有影响 ,但抑制卵母细胞第一极体的排放 ,降低体外受精率 ,并降低卵母细胞的存活率。实验结果提示 ,氯化镍破坏小鼠卵母细胞的减数分裂 ,影响小鼠的生殖功能。     

The effect of a propofol-based sedation technique on cumulative embryo scores, clinical pregnancy rates, and implantation rates in patients undergoing embryo transfers with donor oocytes

Study Objective: To determine the effect, if any, of a propofol-based sedation technique on the reproductive outcomes of patients undergoing embryo transfers with donor oocytes. These ova recipients form a unique subgroup, whose clinical outcomes are unrelated to direct anesthetic effects on their reproductive tracts.

Design: Retrospective chart review.

Setting: A 1200-bed university medical center.

Patients: 117 patients who received fresh embryo transfer cycles between January 1991 and December 1995.

Measurements and Main Results: The anesthesia records of 106 women who donated ova were reviewed for propofol usage during the transvaginal needle aspiration of the ova. The medical records of the 117 patients who received these donated embryos were reviewed for cumulative embryo scores, clinical pregnancy rates, and implantation rates. Fourteen patients received ova from women who were sedated with fentanyl and midazolam during ovum retrievals, while 103 patients received ova from women who had been given fentanyl, midazolam, and propofol in doses of 1.87 mg/kg to 8 mg/kg. The pregnancy rate among all patients who received ova from women who received propofol (44 of 103 = 42.7%) was 14.1 % greater than those whose ovum donors did not receive propofol (4 of 14 = 28.6). 78.6% of both propofol and nonpropofol-exposed groups had cumulative embryo scores of greater than 50. Among patients who became pregnant, 52.3% of propofol-exposed and 50% of nonpropofol-exposed cases had greater than 20% implantation rates.

Conclusions: There is no evidence from our data that the administration of propofol during the aspiration of ovarian follicles for oocyte donation had a negative impact on the oocytes as measured by cumulative embryo scores, probability of a clinical pregnancy, or implantation rate.     

Ultrastructure of the final nuclear maturation of bovine oocytes in vitro

Summary Cumulus-oocyte complexes were obtained from cow ovaries by aspiration from small (1–6 mm in diameter) antral follicles after slaughter. Complexes with a compact multilayered cumulus investment were cultured and subsequently processed for electron microscopy after various periods of culture. By morphological criteria the oocytes could be divided into the following sequence of meiotic stages. The oocyte nucleus I stage was characterized by a spherical nucleus located peripherally in the ooplasm while undulation of the nuclear envelope and initial chromatin condensation was seen at the oocyte nucleus II stage. The oocyte nucleus breakdown stage was characterized by formation of long slender projections from the nuclear envelope in which the envelope doubled back on itself, appearance of dense areas and haphazardly oriented microtubules in the nucleus, marked condensation of the chromatin, and dissolution of the nuclear envelope into irregular vesicles and tubules. The condensed chromatin I stage was characterized by the location of condensed chromatin configuration and uniformly oriented microtubules in a dense area peripherally in the ooplasm while the final condensed chromatin II stage was characterized by a gradual invasion of condensed chromatin configurations into a dense area combined with the presence of the first polar body in the perivitelline space.     

卵母细胞浆内单精子显微注射技术的临床应用

目的:研究卵母细胞浆内单精子显微注射(ICSI)技术的临床应用价值及影响ICSI成功率的因素。方法:对接受ICSI技术治疗的69例男性因素不育、不明原因不孕及体外受精失败患者75个周期的病例资料进行回顾分析。结果:对750个成熟卵母细胞进行ICSI操作,正常受精530个,受精率70.67%;25例临床妊娠,临床妊娠率33.33%。2003～2004年ICSI的受精率和卵裂率均显著高于2001～2002年(68.53%Vs75.86%;89.19%Vs95.45%)。女方三个不同年龄组间的受精率、种植率及临床妊娠率存在显著性差异。体外受精失败组受精率显著下降。结论:ICSI技术是治疗不孕不育症的有效方法,其治疗结局受卵子质量、女方年龄及ICSI操作技术等因素的影响。     

Spontaneous cortical granule release and alteration of zona pellucida properties during and after meiotic maturation of mouse oocytes

Exocytosis of cortical granules (CGs) and the concomitant electron density changes of the zona pellucida (ZP) in the absence of sperm penetration were investigated in mouse oocytes processed with tannic acid containing fixation at various stages during and after maturation. After fusion of the CG membrane with the plasma membrane, the CG contents became very electron-dense, due to tannic acid. CG material is seen to be made up of coarse granular structures which gradually change to fine amorphous structures, which accumulate within the developing perivitelline space (PVS). When the coarse CG material attaches to the ZP, small domains exhibiting higher electron density appeared, and the number of these domains gradually increased. Release of CG was observed from metaphase I through metaphase II. In metaphase I to immediately after ovulation, the higher electron density of ZP and CG release was restricted to the cortical area overlying the meiotic spindle. Finally, the CG-free domain formed itself overlying the meiotic spindle as a result of CG release. However, in oviductal ova, CG release additionally occurred in the hemisphere opposite the spindle. At this stage the entire PVS was well developed and contained numerous fine electron-dense materials. Moreover, the inner half of the ZP increased in electron density as well. This change in electron density of the ZP might be associated with released CG material. These results suggest that the “partial cortical reaction” may play an important role in conditioning the ZP prior to ZP reaction. © 1993 Wiley-Liss, Inc.     

玻璃化冷冻对小鼠卵母细胞和颗粒细胞的影响

目的探讨玻璃化冷冻法保存小鼠卵巢组织对卵泡内卵母细胞和颗粒细胞的影响。方法分别应用两种玻璃化冷冻方案(ED20和EE40)冷冻小鼠卵巢组织,常规组织学检测新鲜和复苏卵巢组织内卵泡形态,同时应用TUNEL方法观察冻融前后卵泡内卵母细胞和颗粒细胞凋亡情况。结果ED20组和EE40组卵泡存活率分别为78.99±5.99%和71.50±5.81%,明显低于新鲜卵巢组织。冻融组织卵泡凋亡率较冷冻前显著升高。ED20组、EE40组和新鲜对照组颗粒细胞凋亡的卵泡比例分别为8.30±2.14%、11.98±2.34%和4.95±1.62%,差异有显著性;而冷冻前后卵母细胞凋亡的卵泡比例无明显差异。结论玻璃化冻融过程对小鼠卵泡中颗粒细胞的损伤较大,而对卵母细胞的影响较小。