体外受精-胚胎移植周期透明带透亮致密患者的受精与临床结局

目的：探讨卵子透明带透亮致密患者行常规体外受精（IVF）或卵胞质内单精子显微注射（ICSI）后受精、胚胎发育情况及临床结局。方法：回顾性分析体外受精过程中卵子出现透明带透亮致密的43个周期,其中IvF周期27例,ICSI周期16例,比较受精率与正常受精比例、胚胎质量、临床妊娠结局。结果：27例IVF周期均未受精,行早期补救ICSI,其中23例完全不受精。补救后IVF总体受精率达64．83％,正常受精率59.32％;16例ICSI周期均受精,受精率和正常受精率分别为85．11％和79．01％,均显著高于IVF组（P〈0．05）。IVF组和ICSI组卵裂率分别为97．96％、95．65％,优质胚胎率分别为52．67％、43．75％,组间差异无统计学意义（P〉0．05）。IVF组妊娠率为37．04％,种植率为33．33％;ICSI组妊娠率为31．25％,种植率为25．00％,组间临床结局相近,无统计学差异（P〉0．05）。结论：透明带透亮致密患者的自然受精能力下降,需要采用ICSI方式授精。常规IVF受精失败后采用早期补救ICSI或直接行ICSI能明显改善透明带透亮致密患者的受精结局,但不能改善妊娠结局。     

精子活动力对卯胞浆内单精子注射结局的影响

目的：探讨精子活动力与ICSI结局的关系。方法：对2001．12—2005．04在本中心完成ICSI治疗的139个周期进行分析。根据卵母细胞形态特征对928个MⅡ期卵进行分级（Ⅰ级、Ⅱ级和Ⅲ级），并分别用可获得的精子行显微穿刺。结果：用A级、B级和C级精子穿刺各级别的MⅡ期卵母细胞后，受精率在三者间无显著性差异，但高于D级精子行ICSI（P〈0．05）。用A、B、C或D级精子行穿刺后一旦使卵受精，卵裂率无明显差异。卵母细胞为Ⅰ级或Ⅱ级时。用A级，B级和C级精子穿刺后，优质胚胎率在三者间无显著性差异，但高于D级精子行ICSI；卵母细胞为Ⅲ时．用A、B级精子行ICSI后，优质胚胎率高于C、D级精子行ICSI（P〈0．05）。结论：精子活动力不影响ICSI后卵裂率；不活动精子行ICSI后受精率和优质胚胎率，总体上较活动精子低。卵母细胞质量较差时，活动精子的活动力等级对优质率胚胎有一定影响。     

两种人工激活剂对卵胞浆内单精子注射后未受精卵母细胞激活效果的比较

目的观察两种不同的人工激活剂对行卵胞浆内单精子注射（ICSI）后未受精卵母细胞的激活作用及效果。方法收集ICSI后24h未受精的人类成熟卵母细胞,分别采用乙醇和离子霉素进行激活处理。16～18h后观察第二极体排出及原核形成情况。激活后正常受精的胚胎继续培养3～5d。统计两组的激活率、原核形成情况、卵裂率及后期胚胎发育情况。结果乙醇处理组的激活率为65．6％（21／32）,离子霉素处理组的激活率为83．7％（36／43）,离子霉素处理组的激活率显著高于乙醇处理组（P〈0．05）;乙醇激活处理后共有10个卵母细胞表现为2PN＋2PB,占被激活卵母细胞的47．6％（10／21）,离子霉素激活处理后有24个卵母细胞表现为2PN＋2PB,占被激活卵母细胞的66．7％（24／36）,两组差异无统计学意义（P〉0．05）;乙醇激活组卵母细胞有4个发育到2～4细胞,2个发育到5～8细胞,1个发育到8细胞以上阶段,获1枚桑葚胚;离子霉素激活组卵母细胞有9个发育到2～4细胞,5个发育到5～8细胞,3个发育到8细胞以上阶段,最终获得2枚囊胚和1枚桑葚胚。结论两种激活剂均能激活ICSI后24h未受精的人类成熟卵母细胞恢复受精,并继续发育成胚胎;离子霉素比乙醇更适宜对ICSI后未受精卵母细胞的激活。     

half-ICSI中IVF受精率的差异对临床结局的影响

目的:比较行half-ICSI治疗中IVF受精率的差异对half-ICSI治疗结局的影响。方法:回顾性分析配偶精液分析处于临界状态,无法排除受精失败可能的患者,以及前次IVF受精失败的患者,采用half-ICSI治疗,并根据IVF部分的受精情况进行分组,其中IVF受精失败(定义为IVF受精率≤25%)的half-ICSI组为研究组,包括IVF受精率为0的完全受精失败half-ICSI组24个周期及IVF受精率低下的half-ICSI组19个周期;而IVF正常受精(定义为IVF受精率≥65%)的half-ICSI组19个周期作为对照组。结果:IVF完全受精失败的half-ICSI组中ICSI的受精率、卵裂率、优质胚胎率、植入率、妊娠率、分娩率、流产率分别为68.94%、95.50%、53.77%、19.23%、33.33%、33.33%、0%,IVF受精率低下的half-ICSI组ICSI的受精率、卵裂率、优质胚胎率、植入率、妊娠率、分娩率、流产率分别为71.43%、94.4%、52.54%、18.75%、36.84%、31.58%、14.29%,与对照组即IVF正常受精的half-ICSI组(分别为81.34%、97.25%、55.66%、21.74%、36.84%、36.84%、0%)相比,除受精率研究组与对照组有显著差异外(P<0.05),其余各率差异均无差异(P>0.05)。3组精液分析也无统计学差异。结论:half-ICSI中IVF受精失败将影响ICSI的受精率,但是对胚胎发育及妊娠结局并无显著影响。     

冻融胚胎移植术用于补救卵胞浆内单精子注射治疗周期的临床价值

目的：探讨冻融胚胎移植在补救卵胞浆内单精子注射（ICSI）治疗周期中的临床价值。方法：回顾分析常规体外受精完全失败的306个周期,根据补救ICSI的时间不同分为6～8h补救ICSI组（A组）和18～20h补救ICSI组（B组）,其中行新鲜胚胎移植且有胚胎冷冻的分别记为AI组（n=108）和BI组（n=51）,随后行冻融胚胎移植的分别记为AⅡ组（n：74）和BⅡ组（n=46）,观察各组的妊娠结局。结果：A组正常受精率、卵裂率、优质胚胎率和临床妊娠率均明显高于B组（P〈0．05）;BⅡ组临床妊娠率和胚胎种植率明显高于BI组（32．61％VS11．76％,15．08％VS5．04％,P〈0．05）;AII组临床妊娠率和胚胎种植率较AI组高（44．59％VS34．26％,21．86％VS19．40％）,但无明显的统计学差异（P〉0．05）。结论：合理的补救时间是影响补救ICSI结局的关键因素,进一步的冻融胚胎移植可以改善补救ICSI治疗周期的妊娠结局。     

钙离子载体A23187联合嘌呤霉素激活行卵母细胞质内单精子注射后未受精卵母细胞的研究

目的观察钙离子载体A23187联合嘌呤霉素对卵母细胞质内单精子注射（ICSI）后未受精卵母细胞的激活作用。方法将体外成熟（IVM）-ICSI和常规ICSI后未受精卵母细胞,按行ICSI后体外培养的时间,分为IVM-ICSI 22h组（33个）、IVM—ICSI 44h组（18个）、ICSI44h组（37个）、ICSI68h组（25个）,分别采用钙离子载体A23187联合嘌呤霉素进行激活处理。应用荧光原位杂交（FISH）技术,对来源于双原核合并第二极体合子的激活胚胎进行性染色体分析。结果钙离子载体A23187联合嘌呤霉素能激活行ICSI后22—68h未受精的卵母细胞。其中IVM-ICSI22h组卵母细胞激活率为88％（29／33）、总卵裂率为62％（18／29）、4细胞阶段胚胎发育率为28％（5／18）,1个胚胎发育到桑椹胚阶段;而IVM-ICSI44h组、ICSI44h组、ICSI68h组的未受精卵母细胞激活率分别为56％（10／18）、65％（24／37）、52％（13／25）;总卵裂率分别为20％（2／10）、42％（10／24）、46％（6／13）,仅ICSI44h组有1个胚胎发育到4细胞阶段。FISH对激活胚胎的分析显示,4个胚胎为XX,9个胚胎为XY。结论钙离子裁体A23187联合嘌呤霉素能有效激活行ICSI失败的卵母细胞;行ICSI后22h内,是对未受精卵母细胞进行辅助激活较为理想的时机。激活的双原核合并第二极体胚胎中有雄原核的存在。     

非男性因素不孕体外受精失败行卵胞浆内单精子注射的结局分析

目的:探讨卵胞浆内单精子注射(ICSI)对非男性因素不孕IVF失败患者治疗结局的影响。方法:回顾性分析由于第一周期常规IVF治疗中卵子完全不受精或受精率≤25%,行补救性ICSI的10个周期(补救性ICSI组),以及因前次受精失败而在随后的治疗周期中采取ICSI方法受精的19个周期(后续性ICSI组)的ICSI治疗结局,并以因男方少弱精子症进行第1次ICSI治疗的133个周期为对照组。结果:后续性ICSI组受精率、植入率、妊娠率和分娩率均高于补救性ICSI组,但差异均无统计学意义(P>0.05)。后续性ICSI组优胚率显著高于补救性ICSI组(P<0.05);补救性ICSI组受精率(48.9%)、优质胚胎率(29.2%)、植入率(0%)、妊娠率(0%)、分娩率(0%)均显著低于对照组(分别为72.1%、46.6%、21.2%、45.1%、39.1%);后续性ICSI组受精率、植入率、妊娠率、分娩率分别为55.4%、8.8%、21.1%、15.8%,均低于对照组(P<0.05或P<0.01)。优质胚胎率后续性ICSI组(44.2%)低于对照组,但无统计学差异。结论:对于非男性因素不孕IVF失败患者,ICSI能避免受精失败,但是受精率以及妊娠结局受到卵母细胞隐匿性异常的影响。     

体外受精失败后补行卵胞浆内单精子注射的临床价值探讨

目的:探讨卵胞浆内单精子注射(ICSI)在体外受精完全失败或受精率低于25%的常规IVF周期中的临床价值。方法:回顾分析2001.01-2004.12在我院生殖医学中心接受常规IVF治疗的35例非男性因素不育患者,取卵后体外受精培养16-18h,发现卵母细胞完全未受精或受精率低于25%,立即行ICSI再授精。结果:在24个常规IVF低于25%的周期中,共有197个未受精卵,其中159个MⅡ期卵,显微注射159个,受精123个,最终形成胚胎96个,受精率为77.4%,卵裂率为78.1%,在22个新鲜移植周期(每周期的移植胚胎由来源于常规体外受精卵和补救ICSI后受精卵的胚胎组成),共有4例临床妊娠;在8个冷冻移植周期中(每周期的移植胚胎完全来源于补救ICSI后的受精卵),有1例临床妊娠。在11个常规IVF完全失败周期中,共有89个未受精卵,其中78个MⅡ期卵,显微注射78个,受精63个,卵裂51个,受精率为80.7%,卵裂率为80.9%,在10个新鲜移植周期(每周期的移植胚胎完全来源于补救ICSI后的受精卵)中共有2例临床妊娠;在2个冷冻移植周期(每周期的移植胚胎完全来源于补救ICSI后的受精卵)中有1例单胎妊娠,妊娠早期流产。胚胎来源于常规体外受精卵和补救ICSI后受精卵的移植周期临床妊娠率为18%;胚胎完全来源于补行ICSI后受精卵的移植周期临床妊娠率为20%。结论:ICSI可作为常规IVF失败后的有效补救措施。     

多囊卵巢综合征妇女体外成熟卵母细胞胚胎慢速冷冻复苏移植结局分析

目的评价未成熟卵母细胞体外成熟（IVM）后形成的卵裂期胚胎经慢速冷冻一解冻后的发育能力。方法将2006年1月至2010年12月北京大学第三医院因多囊卵巢综合征（PCOS）合并不孕症行卵裂期胚胎复苏移植的385例患者分为两组：复苏胚胎来源于体外成熟的卵母细胞组（IVM组,46例）和复苏胚胎来源于常规体内成熟的卵母细胞组（IVF组,339例）。采用慢冻速溶法解冻移植后比较两组患者的临床结局。结果IVM组复苏胚胎243枚,复苏后存活162枚,复苏率为66．67％;IVF组复苏胚胎1605枚,复苏后存活1082枚,复苏率为67．41％,两组比较,差异无统计学意义（P〉0．05）。IVM组患者的临床妊娠率和着床率分别为19．30％（11／57）和10．61％（14／132）,明显低于IvF组临床妊娠率（45．45％,175／385）和着床率（26．14％,240／918;P均〈O．05）。结论体外成熟卵母细胞发育形成的卵裂期胚胎慢速冷冻后临床结局欠佳,可能与冻融前胚胎自身的发育潜力有关。     

34个周期玻璃化冷冻卵母细胞临床应用分析

目的:探讨玻璃化冷冻卵母细胞的临床应用可行性。方法:回顾性分析玻璃化冷冻卵母细胞34个周期,对其复苏率、受精率、卵裂率、临床妊娠率等进行统计分析,并筛选出因男方因素不孕的21个周期(研究组)和同期因男方因素进行新鲜卵细胞浆内单精子注射-胚胎移植(ICSI-ET)的78个周期(对照组)进行比较。结果:298枚玻璃化冻存的卵母细胞复苏后存活278枚,对其中成熟的234枚卵母细胞进行ICSI受精后,受精182枚,形成优质胚胎82枚,移植70枚胚胎,11例患者获得成功分娩(10例单胎,1例双胎)。研究组受精率﹑优质胚胎率比对照组低,差异有统计学意义(P0.05),卵裂率、临床妊娠率、种植率、活产率和流产率差异均无统计学意义(P0.05)。结论:玻璃化冻融卵母细胞的技术可以在有适应证的患者中应用,可获得较好的临床结局。     

Successful pregnancy after ICSI with strontium oocyte activation in low rates of fertilization

Fertilization failure (complete fertilization failure or low fertilization rates) after intracytoplasmic sperm injection (ICSI) can occur in rare cases. In the majority of these cases, the unfertilized oocytes are inactivated. Assisted oocyte activation was applied as a treatment option for a case of low fertilization rate as a clinical trial. A patient with a low fertilization rate (ranging from 0% to 33.3%; mean = 17.0%) after eight previous ICSI cycles at another hospital, was diagnosed with fertilization failure. The most likely cause of fertilization failure was failure of oocyte activation. Therefore, artificial oocyte activation by strontium treatment was combined with ICSI to achieve viable fertilized oocytes. Oocytes were stimulated with strontium (10 mM SrCl(2), 60 min) approximately 30 min after ICSl. Six injected oocytes were stimulated and all were then successfully fertilized. Two blastocysts were transferred into the uterus, resulting in a pregnancy and birth. A second pregnancy was achieved following implantation of two cryopreserved embryos (one blastocyst and one morula). In conclusion, strontium treatment was found to be an effective method for artificial oocyte activation in a case with a low fertilization rate after ICSI.     

对体外受精失败后不同授精时间进行补救ICSI的结局比较

目的:探讨初次体外受精(in vitro fertilization,IVF)失败后不同时间进行补救单精子卵胞浆内注射(intracytoplasmic sperm injection,ICSI)授精对胚胎发育潜力及妊娠结局的影响。方法:将104例完全不受精或低受精率的常规IVF周期分为6 h后行补救ICSI短时授精组(A组,再将A组按补救ICSI时间分为6～8 h的A1组和8.1～12 h的A2组)及晚时授精(22 h后行补救ICSI)组(B组),比较各组间的治疗结局。结果:A组总的卵裂率、可移植胚胎率、优质胚胎率、妊娠率与B组相比差异显著(P<0.05)。A1组与A2组相比,受精率、卵裂率、可移植胚胎率、优质胚胎率无显著差异,妊娠率差异显著(P<0.05)。结论:缩短卵龄有利于补救ICSI结局的改善。     

Rescue intracytoplasmic sperm injection (ICSI)-salvaging in vitro fertilization (IVF) cycles after total or near-total fertilization failure

Objective: To evaluate the effectiveness of delayed oocyte reinsemination by ICSI (rescue ICSI) after total or near-total fertilization failure (≤25%) in IVF.

Design: A retrospective clinical study.

Setting: Non–hospital-based IVF program.

Patient(s): Thirty IVF cycles with total fertilization failure and two cycles with ≤25% initial fertilization.

Main Outcome Measure(s): Fertilization and pregnancy rates after rescue ICSI.

Intervention(s): Rescue ICSI 19–22 hours after initial oocyte insemination.

Result(s): A fertilization rate of 60.2% was achieved with rescue ICSI (141 of 234 oocytes, 29 of 32 patients). Of 30 patients with total fertilization failure, 27 had fresh transfers with rescue ICSI embryos. Two additional patients with ≤25% initial fertilization had subsequent replacement of frozen-thawed rescue ICSI embryos. Six pregnancies resulted, including three singleton, one twin, one missed abortion, and one ectopic pregnancy (20.7%). One of the singleton pregnancies resulted from replacement of four frozen-thawed embryos and is the first known pregnancy achieved from cryopreserved rescue ICSI embryos.

Conclusion(s): Rescue ICSI should be considered in the presence of total or near-total fertilization failure in IVF. Early application of rescue ICSI (19–22 hours after insemination) may be critical for establishing fertilization within an optimal window and producing viable embryos and pregnancies.     

Intracytoplasmic sperm injection as a treatment for unexplained total fertilization failure or low fertilization after conventional in vitro fertilization

OBJECTIVE: To determine whether IVF or intracytoplasmic sperm injection (ICSI) should be the choice of treatment in case of a previous IVF attempt with unexplained total fertilization failure or low fertilization (<25%). DESIGN: Prospective study. SETTING: Leiden University Medical Center. PATIENT(S): Thirty-eight couples undergoing IVF and ICSI on sibling oocytes after a first IVF attempt with total fertilization failure or with low fertilization (<25%). INTERVENTION(S): Performing IVF and ICSI on sibling oocytes. MAIN OUTCOME MEASURE(S): Fertilization and (ongoing) pregnancy rate. RESULT(S): A total of 271 oocytes were collected in 24 oocyte retrievals in the total fertilization failure group. Hundred nine oocytes were randomly allocated to IVF and 12 were fertilized (11%); 162 sibling oocytes were allocated to ICSI and 78 were fertilized (48%). In 8 of the 24 patients fertilization occurred after IVF. The pregnancy rate after transfer of 1 IVF and 1 ICSI embryo (n = 3) was 67% and after the transfer of 2 ICSI embryos (n = 21) this was 52%. In the low fertilization group 169 oocytes were collected in 14 oocyte retrievals. Seventy-two oocytes were randomly allocated to IVF and 16 were fertilized (22%). Ninety-seven sibling oocytes were allocated to ICSI and 58 were fertilized (60%). In 7 of 14 patients fertilization occurred after IVF. The pregnancy rate after the transfer of 1 IVF and 1 ICSI embryo (n = 5) was 80% and after the transfer of 2 ICSI embryos (n = 9) this was 33%. CONCLUSION(S): Performing ICSI on some oocytes of a cohort may avoid total fertilization failures both in patients with a history of total fertilization failure and in patients with a history of low fertilization, as the percentage of fertilization is higher after ICSI compared to IVF and the recurrence of total fertilization failure and low fertilization is high after IVF treatment.     

Pregnancy following calcium ionophore oocyte activation in an oligozoospermia patient with repeated failure of fertilization after ICSI

A successful pregnancy outcome after calcium ionophore A23187 oocyte activation in an infertile couple with a repeated failure of achieving fertilization after ICSI is reported. The secondary infertility couple with oligozoospermia underwent ICSI two times. However, none of the oocytes were fertilized. In the third ICSI attempt, three oocytes after ICSI were activated using calcium ionophore for five minutes. Two of three oocytes thus became fertilized. A successful pregnancy outcome was thereafter achieved with the delivery of a healthy infant without congenital abnormalities. Oocyte activation using calcium ionophore was thus found to be a useful method in a case of repeated failure of fertilization after ICSI.     

The present status of artificial oocyte activation in assisted reproductive technology

Intracytoplasmic sperm injection (ICSI) is the most effective treatment for achieving fertilization in assisted reproductive technology (ART). However, fertilization failure occurs. The incidence of fertilization failure after ICSI is 1–5%. Approximately 50% of fertilization failure cases could be attributed to the abnormality of sperm factor. As the fertilization fails after ICSI using mature sperm, round spermatids and globozoospermia, artificial oocyte activation may provide a means of improving fertilization rates in such cases. The oocyte activation treatments used in clinical research include calcium (Ca) ionophore treatment, electrostimulation and strontium treatment. In terms of the efficiency of oocyte activation, electrostimulation and Ca ionophore gave better outcomes than strontium treatment. Strontium treatment causes Ca²⁺ oscillations in mice, so it has been viewed favorably. However, in human oocytes calcium oscillation has not been observed. The fertilization rate after ICSI was low in the case of globozoospermia and wiht round spermatids. Some cases of pregnancy were achieved by ICSI alone and oocyte activation methods were not essential in these cases. Among the various oocyte activation methods currently used, it should be noted that issues of genetic safety have not been addressed for the combined use of these oocyte activation methods. (Reprod Med Biol 2008; 7 : 133–142)     

甲状腺自身抗体阳性对体外受精-胚胎移植结局的影响

目的：探讨抗甲状腺自身抗体（antithyroid autoantibodies,ATA）阳性对体外受精．胚胎移植泐vitro fertilization and embryo transfer,ⅣF—ET）结局的影响。方法：回顾性分析行IVF／ICSI治疗不孕的患者资料。选取甲状腺抗体阳性但甲状腺功能正常者65例（共71个周期）作为研究组（ATA＋组）,492例甲状腺抗体阴性患者（共512个周期）作为对照组（ATA组）。结果：ATA＋组与ATA-患者的一般情况、促排卵天数、促性腺激素使用总量（Gn总量）、hCG注射日E2水平、获卵数、ICSI受精率、可移植胚胎数和妊娠率,组间比较均无统计学差异（P〉0．05）。ATA＋组IVF受精率（59．73％）、优质胚胎率（26．94％）明显低于ATA-组（70．86％和36．31％）;ATA＋组的流产率（37．50％）明显高于ATA组（17．79％）,差异有统计学意义（P〈0．05）。结论：ATA＋对IvF—ET妊娠结局有不利的影响,因此,对于ATⅣ患者在接受IVF—ET治疗前调节甲状腺自身免疫功能可对妊娠结局有利。