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。     

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。     

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。     

不同类型不明原因不孕患者的体外受籼胎移植助孕结局探讨

目的：探讨不同类型不明原因不孕（UI）夫妇接受IVF-ET助孕治疗的结局。方法：共纳入212个因UI接受IVF-ET助孕的周期进行回顾性分析。所有周期均采用短时受精,在IVF受精后6h,对低受精率（〈30％）和IVF完全受精失败周期中未出现第二极体的成熟卵立即进行早期补救ICSI。结局按原发不孕（A1组,139例）及继发不孕（A2,组73例）分组进行比较。并将所有获得成熟卵的周期再分为IVF受精（B1组,181例）与补救ICSI受精（B2组,26例）进行比较。结果：1．A1组较A2组年轻,但不孕年限更长,MII卵IVF受精率及2PN受精率较低,补救ICSI受精周期率明显较高（17.04％比4.17％,P值〈0.05）。A1组因异常受精、未受精、未卵裂或胚胎质量差导致的无可移植胚胎周期取消率明显高于A2组（11.79％比2.74％,P值〈0.05）,但两组移植周期种植率、临床妊娠率及活产率无显著差异（P值〉0.05）。2．Bl组比B2组MII卵受精率更高（93.82％比84.78％,P值〈0.05）,但其余指标比较无显著差异。结论：不明原因不孕夫妇中原发不孕者更容易出现IVF受精失败或低受精率。采用短时受精及补救ICSI受精后可以获得与继发不孕夫妇相当的IVF-ETII临床结局。     

短时受精后补救ICSI与常规受精失败后补救ICSI的比较

目的探讨在常规IVF-ET受精失败后补救ICSI和短时受精后补救ICSI两种方法在临床中的应用。方法选择常规IVF中受精失败的20个周期行晚期补救ICSI,26个周期行短时受精后补救ICSI,两种方法进行比较。结果早期补救的2PN的受精率,种植率,妊娠率分别为85.28%,13.64%,29.63%,明显高于晚期补救ICSI的50.27%,5%,15.79%（P〈0.01）。结论早期补救ICSI较晚期补救ICSI的2PN的受精率,种植率,妊娠率高,能更好的改善妊娠结局。     

75例常规IVF失败后补救ICSI临床结局分析

目的 对常规IVF周期受精完全失败,行补卵母细胞浆内单精子显微注射(rescue ICSI)的结局分析.方法 回顾分析2005.03-2006.06在我院生殖医学中心接受常规IVF治疗,对卵母细胞完全未受精行ICSI再授精的患者,共75例.在随后治疗中,解冻复苏补救ICSI受精卵的胚胎,共移植10个周期.结果 在75例常规未受精补救ICSI周期中,共有759个未受精卵,其中可注射MⅡ卵有626个,正常受精数332个(受精率53.03%),290个卵裂,形成可移植胚胎199个.在75例补救IcsI周期中,有70例新鲜胚胎移植周期,结果4例临床妊娠(妊娠率为5.71%),均为单胎,分娩4例健康婴儿,2男,2女.解冻复苏补救ICSI受精卵胚胎10周期,结果1例临床,为双活胎(妊娠率为10%),分娩2例健康男婴.结论 ICSI可作为常规IVF失败后所采取的补救措施.     

体外受精-胚胎移植中不同受精方式的妊娠结局分析

目的探讨体外受精-胚胎移植中不同受精方式对妊娠结局的影响。方法回顾性分析2014年1月至12月在本院生殖中心接受IVF/ICSI-ET治疗的733个新鲜移植周期,按受精方式分为常规IVF组(605周期)和ICSI组(128周期),比较两组受精率、卵裂率、有效胚胎率、优胚率、胚胎种植率、临床妊娠率、早期流产率、多胎妊娠率、异位妊娠率及胎儿畸形率。结果两组间受精率、卵裂率、有效胚胎率、优胚率、胚胎种植率、临床妊娠率、早期流产率、多胎妊娠率、异位妊娠率及胎儿畸形率差异均无统计学意义(P0.05)。结论 ICSI技术可取得与常规IVF相似的妊娠结局,是治疗男性不育的有效方法。     

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。     

早期补救卵胞浆内单精子注射在常规体外受精中的临床应用价值探讨

目的探讨早期补救卵泡浆内单精子注射(ICSI)在常规体外受精(IVF)中完全不受精或低受精的临床应用价值。方法回顾性分析2013年3月~2015年5月在本院生殖中心常规IVF治疗受精6h后判断为完全不受精或低受精而行早期补救ICSI的52周期患者,统计其实验室培养指标及临床结局,并与同期因男方因素而直接进行ICSI的127周期进行比较。结果早补救ICSI组的受精率、卵裂率、临床妊娠率及种植率和直接ICSI组比较差异无统计学意义(P0.05),而优胚率、胚胎利用率及囊胚形成率显著低于直接ICSI组,两组差异有统计学意义(P0.05)。结论早补救ICSI可作为常规IVF受精失败的补救措施具有较高的临床应用价值,但由于卵母细胞在体外的培养时间延长导致卵子老化,甚至会影响胚胎的发育潜能。建议对高度怀疑受精障碍尤其高龄(≥37岁)、获卵数≥8患者采用Half-ICSI以提高胚胎利用率和累积妊娠率。     

部分卵胞浆内单精子显微注射112例分析

目的分析本中心行部分卵胞浆内单精子显微注射的病例,探讨half-ICSI的指征的可行性。方法选择2004年1月至2008年12月在本中心行half-ICSI的112个周期,按适应症分三组,A组为男方中度少弱畸精子症,B组为前次IVF受精失败或低下(30%),C组为不明原因不孕,以同期常规IVF治疗的1377个周期为对照,分别比较三组中发生受精失败或低下发生率(受精率30%)情况的比例,以及half-ICSI的效果。结果 half-ICSI的112个周期共获卵1633个,行IVF766个卵,受精率为54.7%,其中完全不受精27个周期,受精低下(30%)11个周期,行ICSI部分867个卵,MII765个,受精645个,受精率84.3%,移植周期临床妊娠率为39.4%(其中IVF完全不受精者临床妊娠率为32.0%)。112个周期有38个周期发生受精失败或低下,发生率为33.9%(38/112),其中A组发生率31.8%,B组50.0%,C组30.8%。对照组发生受精失败或低下率为7.0%(96/1377),结果有显著性差异(P0.05)。结论对存在高危因素的病人在同一治疗周期中行half-ICSI可明显减少因不受精无胚胎移植的情况出现,从而提高妊娠率,又因有IVF胚胎移植,可减少ICSI子代遗传和先天缺陷的风险。     

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。     

Prevention of twin pregnancy after in-vitro fertilization or intracytoplasmic sperm injection based on strict embryo criteria: a prospective randomized clinical trial.

A prospective randomized study comparing single embryo transfer with double embryo transfer after in-vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) was carried out. First, top quality embryo characteristics were delineated by retrospectively analysing embryos resulting in ongoing twins after double embryo transfer. A top quality embryo was characterized by the presence of 4 or 5 blastomeres at day 2 and at least 7 blastomeres on day 3 after insemination, the absence of multinucleated blastomeres and <20% cellular fragments on day 2 and day 3 after fertilization. Using these criteria, a prospective study was conducted in women <34 years of age, who started their first IVF/ICSI cycle. Of 194 eligible patients, 110 agreed to participate of whom 53 produced at least two top quality embryos and were prospectively randomized. In all, 26 single embryo transfers resulted in 17 conceptions, 14 clinical and 10 ongoing pregnancies [implantation rate (IR) = 42.3%; ongoing pregnancy rate (OPR) = 38.5%] with one monozygotic twin; 27 double embryo transfers resulted in 20 ongoing conceptions with six (30%) twins (IR = 48.1%; OPR = 74%). We conclude that by using single embryo transfer and strict embryo criteria, an OPR similar to that in normal fertile couples can be achieved after IVF/ICSI, while limiting the dizygotic twin pregnancy rate to its natural incidence of <1% of all ongoing pregnancies.     

Rescue ICSI of unfertilized oocytes after IVF

BACKGROUND: Failed fertilization after IVF occurs in 10-20% of cycles. Conflicting results of rescue fertilization by ICSI have been reported. We therefore compared the success rate in terms of fertilization and pregnancy of cycles in which rescue ICSI was performed with those from a matched control group of primarily ICSI cycles. METHODS: Unfertilized oocytes from IVF cycles with total fertilization failure where at least four metaphase II oocytes were available were treated by ICSI (group I; n = 120). A matched control group was established with patients undergoing ICSI during the same period (group II; n = 280). RESULTS: Both fertilization rate and the proportion of embryos with four blastomeres on day 2 after ICSI were significantly higher in the control group (P < 0.05). Embryo quality, however, was comparable in both groups. The pregnancy rate in the control group was 25.3% whereas in group I with rescue ICSI, no pregnancy was obtained. CONCLUSIONS: Although unfertilized oocytes after IVF can be fertilized by ICSI, the developmental potential of the ensuing embryos is very poor. Therefore, rescue ICSI after total failure of fertilization is not recommended.     

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.     

Andrology: Successful treatment of severe male factor infertility in 100 consecutive cycles using intracytoplasmic sperm injection

In this report, we present the results of our first 100 consecutivecycles of intracytoplasmic sperm injection (ICSI). Overall,fertilization occurred in 98% of cycles and embryos were transferredin 94% (2.6 embryos per cycle). About 50% of patients had embryosfrozen. The overall fertilization rate was 71%, of which 4%were abnormally fertilized (three pronuclei). A total of 30clinical pregnancies were established (32% per transfer), resultingin 18 singleton, six twin and one triplet ongoing pregnancies.The implantation rate per embryo was 15%. There were no significantdifferences in the fertilization or pregnancy rates betweenpatients Who had only occasional motile spermatozoa in the ejaculate,semen that was too poor for routine in-vitro fertilization (IVF),or who had failed routine IVF and/or subzonal sperm injection(SUZI). A group of 18 patients were treated with both ICSI androutine IVF on their first cycle because of the high likelihoodof failed fertilization due to poor sperm morphology (<20%normal). In this group, ICSI oocytes had a fertilization rateof 76% compared to only 15% for the routine IVF (control) oocytes,and six patients conceived after transfer of ICSI embryos (33%),indicating that ICSI can be used successfully on 50% of theoocytes if fertilization failure is expected. Similarly, patientswho had failed to become pregnant with SUZI achieved excellentresults after ICSI. There were no significant differences betweenICSI and routine IVF in the proportions of grade 1, 2 or 3 embryoson day 3 post-oocyte recovery. In conclusion, we have achievedresults comparable to those reported from Belgium and we havefound that ICSI is universally applicable to all forms of severemale factor infertility. ICSI produces fertilization, pregnancyand freezing rates comparable to routine IVF with normozoospermicsamples and has none of the drawbacks of other assisted fertilizationtechniques.     

Day 2 elective single embryo transfer in clinical practice: better outcome in ICSI cycles

BACKGROUND: Single embryo transfer (SET) after IVF/ICSI has been shown to result in an acceptable pregnancy rate in selected subjects. In our unit, SET is routinely carried out among women under the age of 36 in the first or second treatment cycle when a top-quality embryo is available. In order to define further the selection criteria for SET, we have analysed the outcome of elective SET (eSET), including the cumulative pregnancy rate after frozen embryo transfers, performed in the years 2000-2002 in the Oulu Fertility Center. METHODS: During the study period, a total of 1271 transfers were performed, and in 468 cycles SET (39% of all transfers) was carried out. Of the SET cycles, in 308 cases a top-quality embryo was transferred on day 2 and extra embryos were frozen. Of these eSET cycles, ICSI was carried out in 87 cycles (28%). RESULTS: The overall clinical pregnancy rate per transfer was 34.7% in the eSET cycles. In the eSET ICSI cycles, the clinical pregnancy rate was significantly higher than in the corresponding IVF cycles (50.6 versus 28.5%, P < 0.001). The cumulative pregnancy rate per patient after fresh and frozen embryo transfers was also significantly higher after ICSI (71.2 versus 53.4%, P < 0.01). CONCLUSIONS: A high cumulative pregnancy rate per oocyte retrieval can be achieved after eSET in daily clinical practice. The implantation rate of fresh top-quality embryos in the ICSI cycles was significantly higher than in the IVF cycles, possibly due to more successful selection of the embryo for embryo transfer on day 2 after ICSI. In addition, our data suggest that embryo quality is a more important determinant of outcome than the age of the woman.     

体外受精过程中胚胎污染的临床结局分析（附精液反复污染改行ICSI妊娠一例）

目的对体外受精过程中发生胚胎污染后的临床结局的探讨和分析。方法回顾性分析1997年3月至2007年12月,在本中心接受体外受精-胚胎移植（IVF-ET）治疗1634个周期中,体外受精过程中发生胚胎污染10个周期的临床结局。结果10例胚胎污染周期中移植8个周期18个胚胎,均未妊娠。其中1例精液反复污染,改行卵胞浆内单精子注射（ICSI）,移植3个胚胎,双胎妊娠。结论体外受精过程中发生胚胎污染会造成IVF-ET失败,胚胎污染的病人下一周期可改行ICSI治疗。     

A comparison of four different techniques of assisted hatching

BACKGROUND: Assisted hatching (AH) has been proposed as a means to increase the implantation rate in patients with poor prognosis for pregnancy. The procedure appears to be effective when used selectively. Several different methods for AH have been introduced over the years but comparative studies are lacking. The aim of the current study was to compare retrospectively the efficacy of AH performed with four different methods in patients undergoing IVF or ICSI. METHODS: AH was performed prior to day 3 embryo transfer in 794 IVF/ICSI cycles. Indications for AH were females aged >35 years and/or elevated follicular phase FSH levels, previous failed IVF/ICSI cycles, poor embryo quality, and thick zona pellucida (>15 microm). Assignment to one of the four methods of AH was according to the availability of the particular method during the study period. The study was not randomized. RESULTS: Partial zona dissection was used in 239, acid Tyrode in 191, diode laser in 219 and pronase thinning of the zona pellucida in 145. Mean female age, mean number of previous failed IVF/ICSI cycles, number of oocytes retrieved, fertilization and cleavage rates, good quality embryos and zona thickness on day 3 did not differ between groups. Mean number of embryos transferred, implantation rate, clinical pregnancy rate, and abortion rates were likewise similar. CONCLUSIONS: Selective AH using four different methods yields similar implantation and pregnancy rates.     

Frozen embryo transfers: implications of clinical and embryological factors on the pregnancy outcome

BACKGROUND: Frozen embryo transfers are characterized by impaired pregnancy outcome and increased incidence of pregnancy loss as compared with fresh IVF/ICSI embryo transfers. In this study, we performed a retrospective analysis of clinical and embryological factors that potentially influence the outcome of frozen embryo transfer. METHODS: We reviewed the outcome of 1242 frozen embryo transfers with respect to the age of the woman, the method of fertilization, embryo quality before and after freezing and the number of embryos transferred. RESULTS AND CONCLUSIONS: The pregnancy (positive hCG) and clinical pregnancy rates were 25.8 and 21.1%, respectively. A total of 107 (33.3%) of the 321 pregnancies identified by a positive hCG test miscarried either before (18.4%) or after (15%) the clinical recognition of gestational sac(s). The delivery rate for the frozen embryo transfers analysed was 17.2%. Our data revealed that the delivery rate after frozen embryo transfer was dependent on both the woman's age and the quality of embryos transferred, at the same time being unaffected by IVF/ICSI treatment. In addition, the increased woman's age at IVF/ICSI treatment was identified as the only parameter elevating the biochemical pregnancy rate, whereas the clinical abortion rate was found to be unrelated to the clinical or embryological parameters studied.