卵子第一极体、卵周隙及胞浆形态与ICSI后受精率和胚胎质量的影响

目的从MII期卵子的第一极体、卵周隙和胞浆形态三方面综合探讨卵子质量对卵母细胞浆内单精子注射(ICSI)后的受精率、卵裂率、优胚获得率的影响.方法对ICSI中去除颗粒细胞后的MII期卵子在光镜下从第一极体、卵周隙和胞浆形态三方面综合评价卵子质量并按形态表现分为五级.且分别比较各级卵子的受精率、卵裂率及优胚率.结果Ⅰ级卵子与其他各级含异常形态的卵子比较,受精率、卵裂率和优质胚胎率,均有显著差异(P＜0.01).Ⅰ级卵子有最高的受精率(91.14%)、卵裂率(96.35%)和优质胚胎率(83.91%).形态异常的卵子的受精率、卵裂率和优质胚胎率均有明显的下降趋势.在取卵数≤15个及hCG注射日每个卵泡的血清E2浓度在732～1098pmol/L组中,可获得高比例的Ⅰ级卵子.结论根据MII期卵子的第一极体、卵周隙及胞浆形态三方面综合因素进行的卵子分级是可预测ICSI后受精率、卵裂率及优胚率.     

IVF中GnRH拮抗剂给药对多囊卵巢综合征患者VEGF的影响

目的 探讨IVF—ET中GnRH antagonist治疗多囊卵巢综合征（PCOS）对血清及卵泡液中VEGF的影响。方法 将2006年4月至6月42例临床诊断为PCOS的病人随机分为两组：治疗组18人（GnRH antagonist多荆方案）和对照组24人（GnRH agonist长方案），在超促排卵周期的取卵日留取卵泡液，采用EusA方法检测血清和卵泡液内VEGF的水平，同时比较两组间的获卵数、成熟卵母细胞数、卵裂数、受精率、植入率、临床妊娠率及流产率。结果 两组hCG日血清E2、LH水平及HCG日相对采卵日血清VEGF水平差异有统计学意义（P均〈0．05），着床率和妊娠率经Х^2检验，结果差异有统计学意义（P〈0．05），hCG日卵泡液VEGF水平，两组Gn的用量及用药天数，获卵数，成熟卵母细胞数，受精率，卵裂率相比较均无显著差异。结论 GnRH antagonist治疗PCOS不会改变卵泡液的VGEF水平，不会影响卵泡的发育及卵母细胞的受精率和卵裂率，而妊娠率和着床率均升高，E2水平明显降低可能是影响着床率和妊娠率的主要因素。治疗组OHSS的发生率是下降的。GnRH antagonist用药是安全和高效的。     

HCG日单个卵泡雌激素水平对IVF结局的影响

目的回顾性分析在HCG日单个卵泡雌激素水平对于IVF结局的影响。方法收集分析2009年12月至2010年12月在本中心行IVF/ICSI的不孕患者,所有病人均第一次接受IVF,共161个周期,分为妊娠组与非妊娠组,E2/卵泡<100,100-200,>200组,计算比较各组的一般情况,Gn的用量,促排卵天数,获卵数,受精率,卵裂率,优胚率,妊娠率等,并用分析HCG日血清E2,E2/卵泡(卵泡指HCG日≥14mm的卵泡)与受精率,卵裂率,HCG日LH的相关性。结果 E2/卵泡100-200组与>200组的获卵数,受精率,卵裂率与可用胚率均无统计学差异,而E2/卵泡100-200组的妊娠率明显高于E2/卵泡>200组,两者比较有非常明显的差异(P=0.006)。E2/卵泡与受精率无相关性,与可用胚率呈负相关,与HCG日LH水平呈正相关。结论 HCG日单个卵泡雌激素水平对IVF结局有一定的预测作用,可以作为决定HCG注射时间的重要参考因素。     

电针干预对肾虚型PCOS患者影响机制的研究

目的探讨电针干预对肾虚型多囊卵巢综合征(PCOS)患者体外受精-胚胎移植(IVF-ET)中卵细胞质量及妊娠结局的影响,并探讨其作用机理。方法将接受IVF-ET的肾虚型PCOS患者66例随机分为观察组34例,对照组32例,观察组加用电针干预。观察两组患者血清及卵泡液中干细胞因子(SCF)含量以及周期结局的不同。结果1、观察组治疗后肾虚症状改善明显(P(0.05),受精率、卵裂率、优质胚胎率均高于对照组,差异有显著性(P(0.05)。2、观察组取卵日血清、卵泡液SCF水平明显高于对照组(P(0.05)。妊娠组血清、卵泡液SCF水平高于非妊娠组,差异有显著性(P(0.05)。3、观察组临床妊娠率高于对照组,但差异无统计学意义(P0.05)。4、两组患者年龄、不孕年限、体重指数、基础内分泌、治疗前肾虚症状积分、hCG日子宫内膜厚度、血LH、E2、P水平、获卵数、OHSS发生率、周期取消率均无显著性差异(P0.05)。结论对PCOS患者IVF-ET中进行电针干预,可从整体上调节机体内分泌和卵巢局部微环境,提高了卵子质量和临床妊娠率,取得较高的临床效果。     

Triptorelin和Buserelin对不孕妇女体外受精—胚胎移植疗效的比较

目的 比较GnRH-a制剂Triptorelin和Buserelin的体外受精—胚胎移植治疗效果.方法 185例不孕妇女随机分成两组,A组:85例,黄体期应用Triptorelin(月经第21d皮下一次性注射1.25～1.60mg);B组:100例,黄体期应用Buserelin.两组均用长方案降调节.比较两组的高纯度促卵泡刺激素用药剂量、取卵数、受精率、卵裂率和妊娠率.结果 两组的取卵数、受精率、卵裂率和妊娠率无显著性差异(P＞0.05).A组平均每周期用高纯度卵泡刺激素40.3±18.2支,B组平均每周期用30.2±12.3支,有显著性差异(P＜0.01).结论 Triptorelin和Buserelin的长方案均可获得高质量的成熟卵子,对不孕妇女的体外受精—胚胎移植治疗效果相似.Triptorelin的降调节作用较Buserelin的强.黄体期一次性皮下注射Triptorelin1.25～1.60mg可简化治疗过程,患者易于接受.     

ICSI操作中拆卵时间对胚胎质量及妊娠结局的影响

目的探讨人类卵胞浆内单精子注射（ICSI）技术中,取卵后去除卵周颗粒细胞（拆卵）的适合时间。方法选择09年6月至10年3月男方精液正常、因输卵管因素进入周期的ICSI病例共190对夫妇作为研究对象。随机分为2组,分别在取卵后1～2h（G1）和4～6h（G2）拆卵行ICSI。回顾性分析2组卵子成熟率、受精率、卵裂率、囊胚形成率及妊娠率。结果两组在获卵数、卵子成熟率、受精率及卵裂率方面无统计学差异;妊娠率G2组高于G1组,但无统计学意义（P〉0.5）;两组囊胚形成率差异显著（P〈0.005）,G2组高于G1组。结论卵周颗粒细胞的存在对卵子体外成熟尤其是细胞质的成熟是必须的,取卵后培养4～6h拆卵行ICSI胚胎质量好,囊胚形成率高,有更好的妊娠结局。     

黄体中期长方案及拮抗剂方案对IVF患者卵母细胞质量的影响

目的比较两种控制性超排卵方案对卵母细胞质量的影响。方法回顾性分析2012年1月至2014年7月在北京妇产医院生殖医学科接受黄体中期长方案317例(326个周期)及拮抗剂方案218例(220个周期)治疗的患者,对其卵母细胞质量进行评级,比较两组卵母细胞质量、成熟率、受精率、卵裂率及优质胚胎率等。结果两组患者年龄、不孕年限、BMI、基础FSH、LH、E2水平、Gn总量均无统计学差异(P0.05)。黄体中期长方案组获卵数、成熟卵子数,3级卵子数显著高于拮抗剂组(P0.05);受精数、正常受精数、卵裂数、优胚数均显著高于拮抗剂组(P0.05)。两组的卵母细胞成熟率及各级成熟率、正常受精率、卵裂率、优胚率差异无统计学意义(P0.05)。结论黄体中期长方案组的获卵数、优卵数及优胚数均高于拮抗剂方案组,但两组间成熟卵子率、卵裂率、及优胚率均无差异,两种方案对患者卵母细胞质量影响差异不明显。     

体外受精-胚胎移植患者卵泡液中免疫球蛋白的测定

目的　了解体外受精 -胚胎移植患者卵泡液中是否存在免疫球蛋白 (Ig)及补体C3,及其与所获卵子数、受精率、卵裂率及妊娠成功率的关系。方法　体外受精 -胚胎移植 (IVF -ET)患者 50例 ,分为两组 :妊娠组与非妊娠组 ,利用速率散射比浊法测定每例患者血清及卵泡中的IgG、IgA、IgM、C3水平及记录各个病人的卵子数、受精数、卵裂数。结果　卵泡液中Ig、C3比血液中的显著降低 (P <0 0 1 )且卵泡液中IgA、IgM与血液中IgA、IgM有相关性。卵泡液中Ig、C3与所获卵子数、受精率、卵裂率、妊娠率无相关性。结论　卵泡液中存在免疫球蛋白 (Ig)及补体C3且不影响妊娠的成功。     

Triptorelin和Buserelin对不孕妇女体外受精一胚胎移植疗效的比较

目的　比较GnRH -a制剂Triptorelin和Buserelin的体外受精—胚胎移植治疗效果。方法　 185例不孕妇女随机分成两组 ,A组 :85例 ,黄体期应用Triptorelin(月经第 2 1d皮下一次性注射 1.2 5～ 1.6 0mg) ;B组 :10 0例 ,黄体期应用Buserelin。两组均用长方案降调节。比较两组的高纯度促卵泡刺激素用药剂量、取卵数、受精率、卵裂率和妊娠率。结果　两组的取卵数、受精率、卵裂率和妊娠率无显著性差异 (P >0 .0 5 )。A组平均每周期用高纯度卵泡刺激素 4 0 .3± 18.2支 ,B组平均每周期用 30 .2± 12 .3支 ,有显著性差异 (P <0 .0 1)。结论　Triptorelin和Buserelin的长方案均可获得高质量的成熟卵子 ,对不孕妇女的体外受精—胚胎移植治疗效果相似。Triptorelin的降调节作用较Buserelin的强。黄体期一次性皮下注射Triptorelin1.2 5～ 1.6 0mg可简化治疗过程 ,患者易于接受。     

人卵泡液氨基酸谱影响卵母细胞发育潜能的初步研究

目的分析体外受精（IVF）患者单卵泡液氨基酸谱含量与卵母细胞发育潜能的相关性。方法利用高效液相色谱（HPLC）分析技术检测49份IVF患者优势卵泡的单卵泡液氨基酸谱,并追踪所获卵母细胞发育结局。依据取卵后第三天（D3）胚胎评分分为可用胚胎组（A组）与不可用胚胎组（B组）,比较两组氨基酸谱含量差异。结果①两组患者的年龄、不孕年限、体重指数、基础性激素及用药支数均无统计学差异（P〉0.05）;②A组患者的取卵数、受精率及卵裂率略高于B组,但无统计学意义（P〉0.05）;A组患者的可用胚胎数及可用胚胎率（7.41±4.64、65.15%）显著高于B组（5.00±3.38、49.75%）（P〈0.05）;③A组卵泡液中丙氨酸（Ala）含量（335.86±70.79μmol/ml）明显高于B组（293.56±67.30μmol/ml）（P〈0.05）,其余氨基酸含量无统计学差异（P〉0.05）。④利用ROC曲线确定卵泡液中Ala含量预测可用胚胎形成的临界值为325.4μmol/ml,灵敏度63%,特异度81.8%。结论优势卵泡的发育可在一定程度上反映IVF患者的总体情况,卵泡液中Ala含量可作为预测卵母细胞受精后早期胚胎发育的参考指标之一。     

The effect of propofol anaesthesia on oocyte fertilization and early embryo quality

Propofol, frequently used for i.v. induction of anaesthesia in assisted reproduction procedures, has been suspected of damaging oocytes. Concentrations of propofol have recently been shown to increase in follicular fluid during oocyte retrieval. Our study was designed to assess whether exposure to increasing concentrations of propofol has a measurable effect on in-vitro fertilization, cleavage and embryo development. A cohort of 130 women underwent i.v. anaesthesia using propofol and fentanyl. Time of anaesthesia from i. v. injection of propofol was measured, as were the doses of the two drugs. In 32 women expected to have more than 15 oocytes retrieved, first, middle and last oocytes were cultured separately. The mean time from i.v. injection to first follicle aspiration was 200 s. The mean time for the aspiration of each additional oocyte was 17.6 s. In 10 out of 11 cases where follicular fluid concentrations of propofol were measured, there was an increase from the first to the last follicle, but no difference was found in the ratio of mature to immature oocytes. Nor were any differences found in fertilization, cleavage and embryo cell number. In so far as in-vitro development reflects embryo quality, we conclude that the time elapsed between retrieval of the first and last oocyte does not affect oocyte quality.     

Relationship between fertilization results after intracytoplasmic sperm injection, and intrafollicular steroid, pituitary hormone and cytokine concentrations

Previous studies relating hormone and cytokine concentrations in follicular fluid to oocyte fertilizability were flawed by the uncertainty about the actual oocyte maturity status at the time of recovery and by the possible contribution of the male factor to failures of conventional in-vitro fertilization. This is the first study in which oocyte maturity was assessed immediately after recovery and only mature oocytes were selected for treatment by intracytoplasmic sperm injection. Fertilization outcomes were related to follicular fluid concentrations of 17beta-oestradiol, progesterone, follicle stimulating hormone, luteinizing hormone (LH), growth hormone (GH), prolactin (PRL), interleukin-1 (IL-1) and tumour necrosis factor-alpha (TNF alpha). Those oocytes that subsequently showed normal fertilization were harvested from follicles with higher concentrations of progesterone, GH, PRL, IL-1 and TNF alpha as compared with those of oocytes that failed to fertilize. Among the normally fertilized oocytes, low GH concentrations were associated with the failure of cleavage and with poor morphology of cleaving embryos, whereas rapidly cleaving embryos developed from oocytes recovered from follicles with high concentrations of LH and IL-1. These data suggest important roles for GH, IL-1 and TNF alpha, and of residual LH after pituitary suppression, as positive regulators of the final phase of oocyte intrafollicular development.     

Physiology: Follicular volume and number during in-vitro fertilization: association with oocyte developmental capacity and pregnancy rate

This study examined the effect of the number of follicles aspiratedduring egg retrieval on pregnancy likelihood during an ovulationinduction, in-vitro fertilization (IVF) programme. In addition,the volume of each individual follicle was related to the probabilityof obtaining an oocyte from that follicle. Its capacity to befertilized, the incidence of polyspermic fertilization, thequality of early embryos derived from that oocyte and the influenceof patient age were the other outcomes studied. Large follicularnumber and high mean follicular volume related positively topregnancy outcome. Successful egg retrieval, normal fertilizationand good embryo quality were more likely with increasing individualfollicular volume. A model was constructed to quantify the predictivevalue of follicular fluid volume on the developmental potentialof individual oocytes and embryos derived from that follicle.Successful outcome of IVF is more likely if ovulation inductionresults in many follicles, particularly if they have a highmean volume. Individual oocyte and embryo quality can be tracedback to the follicular level.     

Reduction of progesterone receptor expression in human cumulus cells at the time of oocyte collection during IVF is associated with good embryo quality

BACKGROUND: It has been reported that the progesterone receptor (PR) level is transiently increased within the follicle by LH stimulation and controls cumulus cells in follicles and oocyte maturation. The purpose of this study was to predict developmental competence of human oocytes during IVF via analysis of PR in cumulus cells surrounding mature oocytes. METHODS: Prior to oocyte retrieval, the follicular diameter was measured and follicular fluid was collected from each mature follicle. Cumulus cells were manually separated from the oocyte-cumulus complex under a microscope. PR and PR mRNA were assessed by immunohistochemistry and real-time quantitative polymerase chain reaction (RT-PCR) measurement in human cumulus cells. RESULTS: Immunoreactive PR-A was mainly localized in the cytoplasm and PR-B was localized in the nuclei. There was no significant relationship between PR expression and follicular diameter, follicular fluid concentration of steroids, or LH. There was no significant relationship between expression of PRs and fertilization or cleavage rate. However, PR expression was lower in the good morphology group (blastomeres > or =7 cells with fragmentation > or =5% on day 3) when compared to the other groups (P<0.05). CONCLUSIONS: These results suggest that follicular LH or steroids do not affect PR expression, and full reduction of total PR expression on cumulus cells at the time of oocyte collection is associated with good morphology in human oocytes.     

Propofol anaesthesia for ultrasound guided oocyte retrieval: accumulation of the anaesthetic agent in follicular fluid.

Propofol (2,6-diisopropylphenol, Diprivan, ICI-Pharmaceuticals, Manchester, UK) is widely used either as an adjunct in general anaesthesia or as sole anaesthetic agent by the continuous intravenous route and intermittent bolus injections for minor surgical interventions. For several years, we have been using this kind of anaesthesia in transvaginal oocyte retrieval for in-vitro fertilization (IVF), allowing a completely painless puncture on an out-patient basis. From in-vitro studies on mouse oocytes, it appeared that propofol could be deleterious for fertilization in a dose- and time-dependent manner. We therefore investigated the concentrations of propofol in follicular fluid during oocyte retrieval in women. We measured propofol levels in serum and follicular fluid of nine patients at fixed intervals during ultrasound guided oocyte retrieval. Serum levels fluctuated randomly, due to interference from top-off doses of propofol. In follicular fluid, however, we found a steady increase of propofol levels, which was proportional to the total dose of propofol administered. These data indicate that propofol accumulates in follicular fluid. Although it seems unlikely that propofol as used in the present protocol exerts a clinically significant unfavourable effect on IVF, we suggest that the oocyte retrieval procedure should be kept as short as possible, in order to limit the accumulation of the anaesthetic in follicular fluid.     

Spindle imaging: a new marker for optimal timing of ICSI?

BACKGROUND: The LC Polscope facilitates visualization of the meiotic spindle in human oocyte. This study aimed to investigate meiotic spindle assembly in correlation to time elapsed after HCG administration, and to determine whether spindle imaging may serve to indicate the likelihood of fertilization and embryo cleavage. METHODS: Metaphase II (MII) oocytes from 103 couples who were being treated for male infertility were imaged with the Polscope prior to sperm injection. Spindle imaging was correlated to time elapsed from HCG administration, fertilization rate and embryo cleavage. The main outcome measures were spindle visualization, fertilization and embryo cleavage on day 3. RESULTS: A total of 770 MII oocytes were imaged. A spindle was imaged in a significantly higher number of oocytes from >or=38 h after HCG administration compared with those in the <38 h group (78.1-81.5% versus 61.6%; P < 0.001). The fertilization rate in oocytes with a visible spindle was statistically higher compared with oocytes in which spindle could not be detected (70.4% versus 62.2%; P = 0.035). We found no relationship between spindle imaging and embryo cleavage on day 3. CONCLUSIONS: Spindle imaging, in addition to first polar body appearance, is an accurate indicator for oocyte maturity. We suggest that spindle imaging be performed prior to sperm injection.     

Nitric oxide (NO) and interleukin-1beta (IL-1beta) in follicular fluid and their correlation with fertilization and embryo cleavage

PROBLEM: Using an IVF model, the goal of the study was to investigate the relationship between follicular fluid (ff) NO and IL-1beta levels, as well as their correlation with fertilization of mature oocytes and embryo cleavage rates. METHOD OF STUDY: Follicular fluid was collected from 17 patients at the time of transvaginal oocyte retrieval following controlled ovarian stimulation. Oocytes harvested from these follicles were followed through fertilization and embryo cleavage. The NO metabolites nitrate/nitrite (NO3/NO2) were measured using the Griess reaction as an indirect assessment of NO activity. IL-1beta was measured using a high sensitivity ELISA system (Amersham, UK). The Student's t-test was utilized for unpaired data with the means considered significantly different when P < or = 0.05. RESULTS: Follicular fluid NO3/NO2 levels were significantly lower in follicles containing mature oocytes that fertilized (n = 30; 9.7 +/- 1.0 microM), versus those that did not fertilize (n = 23; 15.4 +/- 2.4 microM; P < 0.05). Follicles that contained oocytes that fertilized and went on to divide beyond the 6 cell stage had significantly lower ff levels of NO3/NO2 (n = 18; 7.5 +/- 0.9 microM), as compared to ff that contained oocytes that did not fertilize or failed to develop beyond the 5 cell stage (n = 35; 14.6 +/- 1.7 microM; P < 0.01). No correlation was found between ff NO3/NO2 levels (n = 28; 13.8 +/- 2.0 microM) and ff IL-1beta levels (n = 28; 0.5 +/- 0.08 pg/mL). An analysis of ff IL-1beta levels in relation to fertilization and embryo cleavage rates revealed no correlation. CONCLUSIONS: Lower ff NO3/NO2 levels at the time of oocyte retrieval are associated with adequate fertilization and embryo cleavage rates. In our IVF model, no correlation was found between ff IL-1beta levels and ff NO3/NO2, fertilization, or embryo cleavage rates.     

High bone morphogenetic protein-15 level in follicular fluid is associated with high quality oocyte and subsequent embryonic development

BACKGROUND: Bone morphogenetic protein-15 (BMP-15) has been shown to influence oocyte maturation and quality. However, no relationship has been established between BMP-15 and oocyte quality/embryonic development in humans. The aim of this study is to investigate BMP-15 level in human follicular fluid (FF) and its possible role in determining oocyte quality and developmental potential. METHODS: A total of 79 oocytes and their corresponding FF from 79 women undergoing ICSI were examined. Individual oocytes were inseminated and subsequently assessed on the basis of their fertilization, cleavage and preimplantation development. BMP-15, FSH, estradiol (E(2)) and progesterone levels of FF were also analysed via the techniques of western blot or radioimmunoassay. RESULTS: Higher FF BMP-15 levels were observed in the fertilized and cleaved groups versus the unfertilized and uncleaved groups, respectively (P < 0.05). The best (Grade I) embryo morphology was associated with higher FF BMP-15 levels than Grade II or III embryos (P < 0.01). A significant positive correlation was found between BMP-15 and E(2) levels in the same follicle. CONCLUSION: The present study demonstrates that the BMP-15 level in FF appears to be a potential factor in predicting oocyte quality and subsequent embryo development, and is correlated with E(2) level, which may additionally be a valuable predictor of oocyte fertilization.     

Antioxidants and reactive oxygen species in follicular fluid of women undergoing IVF: relationship to outcome

BACKGROUND: The role of free radicals and reactive oxygen species (ROS) in female reproductive function is still unclear. The present study was designed to investigate their relationship with ovulation, fertilization and conception. METHODS: Follicular aspirates obtained from women undergoing IVF following controlled ovarian stimulation were evaluated using the ferric reducing antioxidant power (FRAP) assay for baseline total antioxidant capacity (TAC). Both the baseline TAC and the decline in TAC over 72 h (two-point assay) were used as markers of oxygen radical activity. RESULTS: A total of 303 follicular aspirates from 63 women were analysed. Two hundred and eighteen (71.9%) yielded oocytes, 169 (77.5%) of these fertilized and 134 (79.3%) of these embryos survived until the time of embryo transfer. Baseline TAC was no different in follicular fluid whether the follicle contained an oocyte or not, but was significantly higher in fluid from follicles whose oocyte successfully fertilized and significantly lower in fluid from follicles where the resultant embryo survived to transfer. The decline in TAC was lower when the oocytes fertilized and higher in association with embryo viability, but the differences were not statistically significant. CONCLUSIONS: These results provide further evidence that ROS play a role in female reproductive function.