精子DNA碎片对IVF/ICSI结局的影响

目的探讨精子DNA碎片对体外受精/卵胞浆内单精子注射(IVF/ICSI)结局的影响。方法回顾性的分析了行IVF/ICSI治疗的725例不孕夫妇(IVF 502例、ICSI 223例)的临床资料。根据精子DNA碎片指数(DFI)分为正常组(DFI10%)、轻度DFI组(10%≤DFI20%)和重度DFI组(DFI≥20%),分别比较了IVF和ICSI中三组的受精率、卵裂率、可用胚胎率、优质胚胎率和临床妊娠率。结果在IVF周期中,DFI对受精率、卵裂率、可用胚胎率、优质胚胎率和临床妊娠率均无影响(P0.05);在ICSI周期中,DFI对受精率和卵裂率无影响(P0.05),但可用胚胎率和优质胚胎率三组之间有显著性差异(P0.05),临床妊娠率虽然随着DFI的增高而下降,却无显著相关性(P0.05)。结论精子DNA碎片对IVF结局基本无影响,但与ICSI结局中的可用胚胎率和优质胚胎率呈负相关,这可能与受精过程中IVF周期中卵子对精子有自然选择的过程,而ICSI周期中则是人为的选择精子有关。     

获单卵周期体外受精和单精子胞浆内注射对IVF-ET结局的影响

目的取卵日仅获单卵周期常规体外受精（IVF）和单精子胞浆内注射（ICSI）两种受精方式对的IVF—ET结局的影响。方法本研究对92个获单卵周期进行比较，分析了在年龄、激素和子宫内膜厚度无差别的情况下ICSI和IVF的结局。就不同年龄段及不同精液质量时，ICSI和IVF受精率、卵裂率及妊娠率进行比较。结果年龄、激素和子宫内膜厚度无差别时，ICSI的受精率和妊娠率分别为81．6％和12.2％，明显高于IVF组的69．8％和4．7％。不同年龄组，当精子正常时，ICSI组和IVF组受精率和卵裂率无统计学差别，但妊娠率在前者高；精子未达正常标准时，ICSI组的受精率、卵裂率及妊娠率均显著高于IVF组。结论精子质量对选用ICSI还是IVF是重要影响因素，未迭正常标准的精子选用ICSI更好。总体说来，对于获单卵周期，ICSI似乎是更好的受精方式。这次结果也为我们在自然周期选择何种受精方式提供了参考。     

未成熟卵体外培养发育潜能探究

目的体外受精-胚胎移植(In vitro fertilization and embryo transplantation,IVF-ET)周期所获卵子均未成熟时,其体外培养发育潜能探究。方法 25例IVF-ET周期所获卵子均未成熟(未成熟卵149枚),通过实验室体外过夜培养所有未成熟卵子(M1期,142枚;GV期,7枚),培养成熟卵子(M11期,98枚)行体外单精子胞内注射(Intracytoplasmic sperm injection,ICSI)后进行72h体外培养;回顾性分类统计IVF-ET周期中IVF、ICSI和Half ICSI的正常受精率、正常卵裂率和优胚率,比较它们间的差异。结果 IVF周期与Half ICSI周期中的IVF部分在受精率、正常卵裂率和优胚率均无统计学差异;ICSI周期与Half ICSI周期中的ICSI部分在受精率、正常卵裂率和优胚率均无统计学差异;所获卵子均未成熟周期与ICSI周期在受精率和优胚率有统计学差异,在正常卵裂率无统计学差异;所获卵子均未成熟周期与HalfICSI周期中的ICSI部分在受精率和优胚率有统计学差异,在正常卵裂率无统计学差异。结论所获卵子均未成熟周期的未成熟卵子经体外培养能部分成熟,并具有继续发育潜能,但其受精率和优胚率均明显低于体内成熟卵子的相应指标;且他们最终临床结局较差。     

反复体外受精方法失败者卵胞浆内单精于注射预后分析

目的 探讨反复常规体外受精—胚胎移植(IVF—ET)方法失败伴受精不良者行卵胞浆内单精子注射(ICSI)预后。方法 收集1999年10月到2002年8月在我院生殖中心接受常规IVF技术治疗至少1个周期以上失败伴受精率低于50％的23例(31个周期)女性不育患者为研究对象，在以后的治疗周期采用ICSI治疗方法。同时选择2001年10月到2002年8月因男性因素不育而行ICSI的30例(32个周期)患者为对照组，对比两组的临床结果。采用常规超排卵治疗长方案治疗，成熟的卵母细胞行单精子显微注射。结果 两组平均获卵数和受精率、四、六细胞卵裂率无显著差异。但八细胞卵裂率、种植率、临床妊娠率，IVF失败组分别为：26．6％、7．5％、21．4％，男性因素不孕组分别为：35.8％、18％、47.2％。两组有显著性差异。同时比较失败组自身前后两种不同受精方法的实验室情况。IVF的受精率，四、六细胞卵裂率无显著差异。但八细胞卵裂率、种植率、临床妊娠率，IVF失败组分别为：26．6％、7．5％、2l、4％，男性因素不孕组分别为：35．8％、18％、47.2％。两组有显著性差异。同时比较失败组自身前后两种不同受精方法的实验情况。IVF的受精率，四、六、八细胞卵裂率分别为48．1％、35.4％、29．9％、26.8％。ICSI的分别为68.9％、53．7％、29．9％、26.6％。其受精率及四细胞卵裂率有显著差异，六、八细胞卵裂率无差异。结论 ICSI可提高反复IVF-ET失败件受精不良患者受精率，并使部分患者妊娠，但其优质胚胎率、种植率及临床妊娠率相对单纯男性因素行ICSI者较低。提示反复IVF-ET失败伴受精不良者可能存在某种配子异常，显然ICSI可使卵子受精，但配子的异常使胚胎的发育受到损害，降低了种植率和临床妊娠率。     

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。     

血清抗体对体外受精--胚胎移植治疗结局的影响

目的探讨血清抗体抗精子抗体（ASAb）、抗内膜抗体（EmAb）、抗心磷脂抗体（AcA）对体外受精-胚胎移植（IVF-ET）中胚胎着床的影响。方法患者为2001年1月至2002年10月所有行IVF—ET治疗的患者，采用酶联免疫吸附法（EHSA）法检测静脉血ASAb、EmAb、AcA水平，同时观察IVF-ET周期的妊娠结局。筛选出AsAb阳性30周期（30例），EmAb阳性30周期（30例），AcA阳性30周期（30例），做为观察组。抗体阴性30周期（30例）做为对照组，观察120个IVF-ET周期受精率、卵裂率、妊娠率及流产率等。结果30个As址阳性周期受精率64．21％，卵裂率73．64％都低于对照组。30个EmAb阳性周期的妊娠率16．67％，明显低于对照组的11例36．33％。30个AcA阳性周期流产率25％，明显高于对照组1例10％。生化妊娠30个AcA阳性周期37．53％明显高于对照组25％。未妊娠30个EmAb阳性周期最高为31．27％，高于对照组22．07％。生化妊娠与未妊娠者两组比较无显著差异（P〉0．05）。结论妇女体内AsAb、EmAb、AcA可能干扰受精卵的早期发育和早期着床．干扰胚胎发育、胎盘的形成，引起流产等。女性体内的自身抗体也是IVF—ET治疗失败的重要原因之一。     

冷冻复苏的睾丸精子对ICSI助孕结局的影响

目的探讨冷冻复苏的睾丸精子对ICSI助孕结局的影响。方法回顾性分析2014年3月至2017年5月因男方梗阻性无精子症(OA)或非梗阻性无精子症(NOA)在我院采用细针睾丸穿刺抽吸精子行ICSI助孕的209个周期的临床资料,新鲜睾丸精子(新鲜睾丸精子组)用于174个周期,冷冻复苏的睾丸精子(冻精组)用于35个周期。比较两组的一般资料、受精率、卵裂率、优质胚胎率、临床妊娠率以及种植率。结果两组的一般资料比较无统计学差异(P0.05)。冻精组与新鲜睾丸精子组行ICSI助孕后的受精率、卵裂率、优质胚胎率、临床妊娠率及种植率比较均无统计学差异(P0.05)。结论冷冻复苏的睾丸精子对ICSI助孕结局并无显著影响。     

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.     

Diff-Quik染色法评估精子形态与体外受精胚胎质量的关系

目的探讨Diff-Quik染色法评估精子形态与体外受精胚胎质量的关系。方法在76个IVF周期中,采用Diff-Quik染色后评估精子畸形率,并分析其与IVF受精率、卵裂率、优质胚胎率之间的关系。结果Diff-Quik染色后精子畸形率与卵母细胞受精率呈负相关(P0.05),与卵裂率、优质胚胎率之间关系不显著(P0.05)。结论Diff-Quik染色后精子畸形率越高,受精率越低,但精子形态与胚胎早期卵裂速度及胚胎形态间无明显相关。     

助孕方式对卵巢低反应患者治疗结局的影响

卵巢低反应患者在体外受精控制性超排卵助孕治疗中发生率9％-24％^[1]，由于获卵数少，许多中心为防止常规体外受精-胚胎移植（IVF—ET）不受精，而行卵胞浆内单精子注射（ICSI）治疗，以改善该患者的助孕结局。本文探讨助孕方式（既选择IVF或ICSI）对卵巢低反应患者治疗结局的影响。     

Effect of in-utero diethylstilboestrol exposure on human oocyte quality and fertilization in a programme of in-vitro fertilization.

Genital tract abnormalities and adverse pregnancy outcome are well known in women exposed in utero to diethylstilboestrol (DES). Data about adverse reproductive performance in women exposed to DES have been published, including controversial reports of menstrual dysfunction, poor responses after ovarian stimulation, oocyte maturation and fertilization abnormalities. We compared oocyte quality, in-vitro fertilization results and embryo quality for women exposed in utero to DES with a control group. Between 1989 and 1996, 56 DES-exposed women who had 125 in-vitro fertilization (IVF) attempts were retrospectively compared to a control group of 45 women with tubal disease, who underwent 73 IVF attempts. Couples suffering from male infertility were excluded. The parameters compared were oocyte quality (maturation abnormalities, immature oocyte, mature oocyte), fertilization and cleavage rate (per treated and metaphase II oocytes), and embryo quality (number and grade). We found no significant difference in oocyte maturational status, fertilization rates, cleavage rates, embryo quality and development between DES-exposed subjects and control subjects. These results suggest that in-utero exposure to DES has no significant influence on oocyte quality and fertilization ability as judged during IVF attempts.     

The influence of different types of antibodies on in vitro fertilization results

PROBLEM: The influence of anti-sperm (ASA), anti-phospholipid (APA), and antizonal (AZA) antibodies on in vitro fertilization (IVF) results and the need for intracytoplasmic sperm injection (ICSI) were assessed. METHOD OF STUDY: Forty-four couples with infertility of immunologic origin were investigated. ASA in serum and ovulatory mucus were studied by a tray agglutination test (TAT) and indirect mixed anti-globulin reaction test (MAR) test, AZA were studied by passive hemagglutination and commercial enzyme-linked immunosorbent assay (ELISA; BioGen, Germany), and APA were tested by ELISAs in immunoglobulin isotypes IgG and IgM. RESULTS: Because of failed or very low fertilization after standard IVF in the previous cycle, ICSI had to be used in five out of 15 cases with ASA (33.3%), in 16 out of 18 couples with AZA (89.4%), and in only one case if APA were present (9%). Clinical pregnancy rate was 60% in cases with ASA, 38.5% with AZA, and 27.3% per embryo transfer (ET) if APA were detected. CONCLUSIONS: Immunologic infertility can be treated by IVF with very good results. The most important group are women with AZA, in whom IVF ICSI without any delay is recommended.     

Low pregnancy rate is achieved in patients treated with intracytoplasmic sperm injection due to previous low or failed fertilization in in-vitro fertilization

The main indications for intracytoplasmic sperm injection (ICSI) are severe male factor and fertilization failure or a low fertilization rate in previous in-vitro fertilization (IVF) treatments. The fertilization and pregnancy rates after ICSI, however, are seldom reported separately for these two different indications. The aim of this study was to compare the treatment outcome and pregnancy rate after ICSI between 65 patients with previous failed fertilization or a low fertilization rate without male factor, and 219 patients with a primary male factor. From the 2726 oocytes collected, 2087 (77%) were micro-injected and 1355 (65%) achieved normal fertilization. The oocyte fertilization rate was similar in the group with previous failed fertilization or a low fertilization rate and the group with a male factor (65 and 65% respectively), as was the cleavage rate of normally fertilized oocytes (92 and 94% respectively). Despite the similar fertilization and cleavage rates and the similar number and morphological quality of embryos transferred in both groups, the pregnancy rate was significantly lower (P < 0.05) in the group with previous failed fertilization or a low fertilization rate than in the group with a male factor (19.6 versus 33.5% respectively; 95% confidence intervals for the difference, 2-26%). The implantation rate was also lower (P = 0.01) in patients with previous failed fertilization or a low fertilization rate (9.6%) than in the group with a male factor (19.5%). We conclude that patients with previous failed fertilization or a low fertilization rate in standard IVF without male factor have a significantly smaller chance of becoming pregnant after subsequent ICSI than patients with a primary male factor. This poor outcome probably reflects intrinsic oocyte defects not bypassed by ICSI.      

卵胞浆内单精子注射在男性与非男性不育患者中的应用

目的比较卵胞浆内单精子注射（ICSI）在男性不育与非男性不育患者中的助孕结局。方法回顾性分析2009年1月至2010年12月在我院接受ICSI治疗109个周期,将其分组：A组：男性不育73周期,包括射精组43个周期,手术抽吸精组30个周期,B组：非男性不育36周期,均为射精组。结果①A组：手术抽吸精组中女方年龄、平均获卵数与B组比较结果有统计学意义,P〈0.05;②射精组与手术抽吸精组：两组的正常受精率、卵裂率、优胚率、种植率、妊娠率比较,结果无统计学差异,P〉0.05;③B组中移植（ET）胚胎数、种植率、妊娠率、流产率与A组相比较,结果有统计学差异,P〈0.05;与正常受精率、卵裂率、优胚率,结果比较无统计学差异,P〉0.05。结论①ICSI仍是治疗男性因素的首要指证;②取精方式对妊娠率无影响;③对非男性因素,能获得较高的受精率,有临床治疗价值。     

常规IVF受精失败补救ICSI后行冻融胚胎移植的临床价值

目的探讨冻融胚胎移植在常规体外受精(IVF)失败后补救卵胞浆内单精子注射(L-ICSI)中的应用价值。方法在12个常规体外受精失败周期中应用ICSI对未受精的MⅡ期卵子进行显微授精,将获得的优质胚胎进行冷冻,再择期行冻融胚胎移植。结果对93个未受精的MⅡ卵子接受L-ICSI,受精63枚,受精率为67.7%(63/93),异常受精3枚(2枚1PN,1枚3PN),57个正常受精卵发生卵裂,卵裂率为95.0%(57/60),优质胚胎率为43.9%(25/57),10例患者冷冻胚胎25枚,其中4例采用程序化冷冻,6例采用玻璃化冷冻。9个患者行冻融胚胎移植,共移植胚胎18枚(其中解冻后胚胎碎裂死亡5枚),其中1个周期因冻融后2个胚胎碎裂放弃移植,2例获得临床妊娠,1例分娩出正常婴儿,1例正在妊娠中,临床妊娠率为22.2%。结论 ICSI可使常规体外受精失败的卵子再受精,冻融胚胎移植可以解决胚胎与子宫内膜不同步的问题,获得相对满意的临床结局,具有一定的应用价值。     

Outcome of conventional IVF and ICSI on sibling oocytes in mild male factor infertility

BACKGROUND: Decisions concerning the treatment choice for assisted reproduction (IVF or ICSI) are usually made after the evaluation of male fertility factors, or after taking into account the results of previous IVF attempts. There are no widely accepted criteria, so decisions for couples with male subfertility are often empirical and may lead to complete fertilization failure after IVF, or to the unnecessary use of ICSI. METHODS: A study was conducted in which half the oocytes from each of 58 couples with moderate oligo +/- astheno +/- teratozoospermia were inseminated (conventional IVF) and the other half microinjected (ICSI). The technique used for subsequent cycles depended on the results of the first cycle. RESULTS: Nineteen of the 58 IVF/ICSI attempts resulted in fertilization after ICSI only (32.8%) and 39 in fertilization after IVF and ICSI (67.2%). For patients with oocyte fertilization only after ICSI, 61.5% of the oocytes microinjected were fertilized. A mean of 2.2 embryos per patient were transferred, leading to eight clinical pregnancies (42.1%).The implantation rate was 21.4%. All subsequent cycles were carried out with ICSI. Couples with oocyte fertilization after both IVF and ICSI had slightly better semen characteristics than those with oocyte fertilization only after ICSI, but this difference was not significant. Overall, no statistically significant difference was observed between IVF and ICSI in sibling oocytes for any of the variables studied: fertilization rate, embryo morphology and rates of development, pregnancy and implantation. Although only small numbers of oocytes or embryos were available for each couple, six couples had lower fertilization rates after IVF and eight had lower embryo quality after IVF. Eight patients had lower sperm quality in the second cycle, and only seven couples underwent subsequent IVF cycles. CONCLUSIONS: This strategy enabled us to avoid 32.8% of complete fertilization failures after IVF, but not to decrease significantly the number of ICSI attempts in subsequent cycles. However, the uncertainties concerning the safety of ICSI suggest that ICSI should be used cautiously and judiciously.     

Sperm DNA fragmentation decreases the pregnancy rate in an assisted reproductive technique

BACKGROUND: Standard sperm characteristics are poor predictors of the outcome of IVF treatments. On the contrary, sperm genome quality has been emphasized for several years as playing a major role in early embryogenesis, thus in the success of IVF attempt. METHODS: Sperm DNA fragmentation from a selected group of 104 couples undergoing assisted reproductive techniques (ART) (IVF: n = 50; and ICSI: n = 54) was measured by TUNEL assay and correlated with semen and ART outcomes. RESULTS: A negative correlation was found between sperm characteristics and the proportion of sperm showing DNA fragmentation. For fragmentation >10%, a significant decrease of the fertilization rate was observed. No correlation was found between sperm DNA fragmentation and embryo quality. A high proportion of sperm with fragmented DNA was a pejorative factor to obtain pregnancies when ICSI was performed, but there was no relationship when conventional IVF was performed. CONCLUSIONS: The proportion of sperm with DNA fragmentation appears to be potentially useful as a predictor of ICSI outcome, whereas embryo quality based on morphological criteria, appeared unaffected by DNA fragmentation.     

GnRH拮抗剂与GnRH激动剂短方案IVF-ET结局的比较

目的比较GnRH antagonist与GnR Hagonist短方案的IVF-ET结局。方法2006年8月至2007年8月GnR Hantagonist治疗组54人和GnR Hagonist短方案对照组132人,记录促性腺激素的用量及其用药天数、hCG日子宫内膜厚度和激素水平、获卵数、受精率、卵裂率、优胚率、妊娠率和OHSS发生率等指标。结果两组促性腺激素的用量及其用药天数、获卵数、受精率、卵裂率、着床率和妊娠率相比较均无显著差异（P〉0.05）。GnR Hantagonist组在hCG日激素水平低,与对照组比较其差异有统计学意义。结论行GnR Hantagonist方案IVF-ET助孕治疗与传统的GnR Hagonist短方案比较,其hCG日雌激素水平下降可能是OHSS发生率显著下降的主要因素;但卵泡的发育、卵母细胞的受精率、卵裂率及妊娠率和着床率均不受影响。GnR Hantagonist的使用为IVF-ET助孕药物提供了一种新的选择。     

常规体外受精失败后行补救性卵胞质内单精子注射的临床总结

目的回顾分析常规IVF受精完全失败在临床中的发生情况及受精失败后应用补救性卵母细胞内单精子注射（ICSI）的临床结局。方法总结1999年5月-2009年5月,在我中心行常规IVF患者中,受精完全失败的发生情况。回顾性分析了2004年1月-2009年5月,因受精完全失败行补救性ICSI后的临床结局。结果1999年至2009年5月,我中心共行常规IVF8359个周期,其中,受精完全失败232个周期,发生率为2．78％。2004年1月至2009年5月,共184个周期受精完全失败,资料完整,对其中115个周期共838个MII期卵子行补救性ICSI,其受精率、卵裂率与优质胚胎率分别为71.0％、93．78％和51．79％;临床妊娠率为11％,获得活产儿5个。结论补救性ICSI能获得较高的受精率和卵裂率,降低周期取消率,但并没有明显增加活产儿出生数,改善妊娠结局。     

IVF-ET患者卵巢颗粒细胞COX-2的表达与妊娠结局的相关性研究

目的探讨体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)患者卵巢颗粒细胞中环氧合酶-2(cyclooxygenase-2,COX-2)的表达,分析其与妊娠结局的关系。方法荧光半定量real-time PCR检测48例行IVF-ET患者卵巢颗粒细胞中COX-2的mRNA表达,分析COX-2与注射hCG日血清雌二醇(estradiol,E_2)水平、获卵率、受精率、优质胚胎率及妊娠率的关系。结果卵巢颗粒细胞COX-2的表达量与hCG日血清E_2水平及获卵率无关,与受精率、优质胚胎率及妊娠率相关。结论 PCOS患者卵巢颗粒细胞COX-2的表达与胚胎质量相关,研究卵巢颗粒细胞的COX-2的表达可以作为预测IVF-ET妊娠结局指标之一。