经皮穿刺附睾吸取精子卵浆内注射受精治疗不育

本文应用经皮穿刺附睾吸精子（ｐｅｒｃｕｔａｎｅｏｕｓｅｐｉｄｉｄｙｍａｌｓｐｅｒｍａｓｐｉｒａｔｉｏｎ，ＰＥＳＡ）经卵细胞浆内显微注射技术（ｉｎｔｒａｃｙｔｏｐｌａｓｍｉｃｓｐｅｒｍｉｎｊｅｃｔｉｏｎ，ＩＣＳＩ），对因精道不通致不育的８对夫妇进行治疗。按ＩＶＦ常规促超排卵方案及阴道Ｂ超取卵，共８个周期。ＰＥＳＡ有３例为输精管绝育术后吻合失败者，５例为先天双侧输精管缺如（ＣＢＡＶＤ），吸出精子活率１～８０％，精子密度２～８２×１０６／ｍｌ，共获６９个成熟卵母细胞，ＩＣＳＩ后７个卵子损伤，６２个存活，其中可见２原核正常受精的５０个，正常受精率为８０．６４％，至１９９８年８月已有５例妊娠，临床妊娠率为６２．５％，出生１例健康男婴，发育正常。结果显示，ＰＥＳＡ结合ＩＣＳＩ技术治疗精道不通不育，是一种有效的治疗方法。     

人卵细胞冷冻保存后经显微注射授精的初步研究

建立人卵细胞冷冻保存以及融解后受精和发育颇具实用价值。本文采用慢冷、快融法，结合使用丙二醇和蔗糖作为冷冻保护剂，对２８５个临床ＩＶＦ未受精卵进行冷冻保存，３天至４０天时间不等。冷冻存活率为５４．７３％，其中未成熟卵子５１．７２％，成熟卵子５５．５０％，二者无显著性差异（Ｐ＞０．０５）。冷冻损伤的卵子中，细胞膜损坏为９６．８９％，透明带破裂为１３．１８％。选择冻融后形态结构正常的１２６个成熟卵子进行胞浆内单个精子注射（ＩＣＳＩ），受精率达４２．０６％，其中５０．９４％受精卵子进一步发育成２～８细胞胚胎。来源于常规ＩＶＦ未受精卵的再次受精率为４０．７４％，来源于ＩＣＳＩ未受精卵的再次受精率为３６．７３％。作者认为，必要时可以借助显微操作技术进行胚胎活检，了解部分染色体畸变情况，使卵子冷冻保存技术安全地应用于临床。     

精子显微授精技术的初步探讨

本文用透明带下精子显微注射法（ＳＵＺＩ）及胞质内精子显微注射法（ＩＣＳＩ）进行小鼠显微授精的可行性及影响因素的研究。其中ＳＵＺＩ法的受精率达２９．２％，卵裂率为４２．８％，移植后出生了两只健康仔鼠。小鼠卵对ＩＣＳＩ法操作耐受差，用ＩＣＳＩ法注射后移植卵未有仔鼠出生。本研究建立了透明带下精子显微注射授精的动物模型，为精子显微注射法应用于临床奠定了基础。     

卵母细胞单精子显微注射治疗男性因素及不明原因不育

目的：研究应用卵母细胞单精子显微注射（ｉｎｔｒａｃｙｔｏｐｌａｓｍｉｃｓｐｅｒｍｉｎｊｅｃｔｉｏｎ，ＩＣＳＩ）技术，治疗因严重男性因素及不明原因引起不育患者的价值。方法：于１９９４年１２月，应用ＩＣＳＩ技术治疗３１例严重少、弱、畸精症及不明原因不育夫妇。按常规超排卵治疗，经阴道Ｂ超介导取卵，行成熟卵母细胞单精子显微注射受精。结果：２３７个成熟卵母细胞中，２１３个卵子存活，其中正常受精卵子１３２个，正常受精率为６２．０％。至１９９６年４月３０日，已有８例临床妊娠，临床妊娠率为２５．８％，其中１例在１９９６年１０月３日，于妊娠３９周＋３顺利分娩一婴儿。结论：ＩＣＳＩ技术用于治疗严重少弱畸精症及不明原因不育夫妇是适宜的。     

体外培养24小时受精失败卵母细胞单精子显微注射再授精的观察

目的 探讨采用单精子显微注射（ＩＣＳＩ）技术处理体外培养２４ｈ后常规体外受精（ＩＶＦ）失败卵母细胞的效果。方法 对１９９７年６月至１９９８年１０月在我院生殖医学研究中心接受常规ＩＶＦ治疗的１７例非男性因素不育患者,采用常规超排卵方案治疗,经阴道Ｂ超介导取卵,共获卵１７９个,体外培养２４ｈ,所有卵母细胞在常规ＩＶＦ失败后,采用ＩＣＳＩ再授精。结果 共有１３０个卵母细胞常规ＩＶＦ失败后进行ＩＣＳＩ再授     

人类胚胎辅助孵化技术和应用概况

人类胚胎辅助孵化技术和应用概况李善国综述邵敬於审校（上海市第一妇婴保健院，上海，２０００４０）由于培养条件改善和授精技术发展，人类体外受精（ＩＶＦ）已达到比较理想的结果。据报道，目前常规ＩＶＦ受精率为６９～９７％［１］，卵子胞浆内单精子注射（ＩＣＳＩ...     

体外受精-胚胎移植过程中多原核孕卵生成的影响因素

目的 探讨体外受精－胚胎移植（ＩＶＦ－ＥＴ）过程中多原核孕卵生成的影响因素及降低该类异常受精的方法。方法 应用多因素Ｌｏｇｉｓｔｉｃ回归方法,分析１４５个ＩＶＦ－ＥＴ周期、１１６８个卵细胞的资料,研究多原核孕卵生成率与夫妇年龄,促超排卵方案,穿刺卵泡数,获取卵细胞数,授精前孵育时间,精液精子与授精精子质量和数量以及卵细胞受精率的关系。结果 成熟卵多原核孕卵生成率与女方年龄和可穿刺卵泡数呈显著性负相     

ZP3对男性生育力的诊断及应用

受精是一个精卵相互作用严格有序而又协调发育的复杂过程，它必须具备三个条件：（1）有正常的生殖细胞（精子和卵子）；（2）精子和卵子能正常结合成受精卵；（3）受精卵能正常着床和发育。不孕不育的原因最常见的是精子或卵子有缺陷，或生殖细胞的遗传物质异常，导致受精失败，或胚胎不能正常发育而流产。临床资料表明，在男女不孕不育中，女方原因占50％，男方原因占40％，男女双方都有问题占10％。     

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。     

体外受精治疗周期中出现过多的多原核合子5例分析

在体外受精中异常受精的现象，已经引起越来越多胚胎学家的关注。在常规体外受精（IVF）中多原核一般认为是由于多精受精引起的，而对于卵胞浆内单精子注射（ICSI），研究者则认为多原核的形成是由于第二极体不能排出，也可能是二倍体的精子或卵母细胞参与了受精。文献报道在体外受精周期中，受精卵中出现多原核的比例约为3％-10％。然而在一些体外受精治疗周期中，受精卵中存在过多多原核形成的现象。本文对体外受精（IVF和ICSI）治疗周期中，部分患者的大部分受精卵中出现多原核的现象进行汇总分析，以更好地了解多原核形成的原因及其机制，为临床上体外受精方法的选择提供依据。     

Complete failed fertilization after intracytoplasmic sperm injection--analysis of 10 years' data

OBJECTIVE: To investigate incidence and causes of complete failed fertilization after intracytoplasmic sperm injection (ICSI) in a tertiary care facility. METHODS: A total of 1,779 cycles between February 1994 and December 2003 were analyzed. Study parameters were female age, infertility diagnosis, ovarian stimulation protocol, estradiol level on day of hCG administration, number of follicles, number of oocytes retrieved, number of oocytes injected, and semen parameters. RESULTS: Complete failed fertilization occurred in 23 cycles (1.29%) involving a total of 85 oocytes injected. Infertility causes among patients with failed fertilization included unexplained (43.6%), male factor (26%), presence of more than one factor (17.4%), hysterectomy (4.4%), premature ovarian failure (4.3%), and advanced age (4.3%). In 12 cycles (52%), fewer than 5 follicles were present. In three (13%) cycles, no mature (MII) oocyte was available and in 61% (14/23) fewer than 3 MII oocytes were available for ICSI. Immotile sperm was used for ICSI in 5 cycles (21.7%). The source of sperm in 17 (74%) cycles was from ejaculate, in 4 cycles from testicular aspiration (TESA), one from percutaneous epididymal sperm aspiration (PESA) and one from retrograde ejaculation. CONCLUSIONS: Our data indicate that major contributing factors to failed fertilization after intracytoplasmic sperm injection are number of MII oocytes retrieved and availability of viable sperm for injection. Although the incidence of complete failed fertilization is not remarkable, it may increase with increasing patient age and a lower number of follicles.     

Management of long-standing unexplained infertility: A prospective study.

OBJECTIVE: Our purpose was to evaluate a protocol for the management of long-standing unexplained infertility. STUDY DESIGN: A prospective study was conducted at an in vitro fertilization-embryo transfer center with 485 patients with the diagnosis of long-standing unexplained infertility. Patients were treated by means of controlled ovarian hyperstimulation and intrauterine insemination for a maximum of 3 cycles. Those who did not become pregnant underwent in vitro fertilization and intracytoplasmic sperm injection on sibling oocytes. RESULTS: Among 485 patients, 921 cycles of controlled ovarian hyperstimulation and intrauterine insemination were performed. The pregnancy rate was 15.7% per cycle and 29.8% per patient. Among those who did not become pregnant, 131 patients underwent in vitro fertilization and intracytoplasmic sperm injection into sibling oocytes; 48 women became pregnant (36.7%). There was total failure of the in vitro fertilization oocytes in 23 cycles (17.6%). CONCLUSION: Management of long-standing unexplained infertility with controlled ovarian hyperstimulation and intrauterine insemination resulted in a reasonable pregnancy rate. Treatment by means of in vitro fertilization and intracytoplasmic sperm injection into sibling oocytes among patients who did not become pregnant resulted in a good pregnancy rate. The overall results showed that the proposed protocol for management of unexplained infertility is satisfactory.     

Chemical activation of oocytes in different types of sperm abnormalities in cases of low or failed fertilization after ICSI: a prospective pilot study

This study evaluated the influence of chemical activation of oocytes in cases with abnormal sperm morphology and previous low intracytoplasmic sperm injection (ICSI) fertilization rate. Fertilization, early cleavage and embryo quality were assessed according to individual abnormal sperm morphology with or without chemical activation of oocytes after ICSI in 56 consecutive cycles with different types of abnormal sperm morphology (A = amorphous head, B = tapered head, C = bent neck) having a history of previous failed or low fertilization rate. Chemical activation significantly increased fertilization rate in subgroup A (82.7 versus 36.7%, P = 0.0008) and subgroup B (81.7 versus 39.3%, P = 0.005), but not in subgroup C (48.2 versus 49.4%). There was no effect on early cleavage (25 h after injection) or on the number of good embryos on day 2 (48 h after injection) in the different subgroups. Chemical activation of oocytes after ICSI in selected cases with previous history of failed or low fertilization rate and 100% sperm abnormality may increase fertilization rate in some sperm morphological abnormalities (amorphous head and tapered head) but not in others (bent neck). However, there was no significant effect on early cleavage or number of good embryos in all subgroups.     

In vitro oocyte maturation and subsequent delayed fertilization is associated with increased embryo aneuploidy

Failed fertilization and the appearance of immature oocytes are common in IVF practice; rescue intracytoplasmic sperm injection can be used as a therapy. However, this study indicates that embryos created after in vitro maturation and delayed intracytoplasmic sperm injection contain an increase in aneuploidy (79.7%) over control embryos (60.5%). Therefore, patients should be informed of the possible risk when presented with delayed intracytoplasmic sperm injection.     

A strategy for treatment of couples with unexplained infertility who failed to conceive after intrauterine insemination

Couples with unexplained infertility treated unsuccessfully with intrauterine insemination often receive further treatment with IVF or intracytoplasmic sperm injection (ICSI). The aim of this study was to evaluate the frequency of fertilization and fertilization failure with respect to the method of fertilization used, when half of the sibling oocytes were fertilized by conventional IVF and insemination and the remainder by ICSI. Included was the first IVF/ICSI treatment of 248 unexplained infertile couples who had failed to conceive after three IUI cycles. An overall pregnancy rate per embryo transfer of 57% was observed. A significantly better fertilization rate was obtained after ICSI as compared with IVF (68 versus 46%) (P < 0.005), and total fertilization failure following ICSI and IVF treatment was seen in 4.4 and 25% of the cycles respectively. The group who experienced total fertilization failure after IVF had normal semen parameters, although significantly lower sperm concentration and motility as compared with the entire study group. Transfer of their ICSI-fertilized oocytes subsequently resulted in a pregnancy rate of 49.2% per transfer. The policy of splitting the sibling oocytes can effectively minimize complete fertilization failure while maintaining high chances of achieving a pregnancy. At the same time, the optimal fertilization method for subsequent treatment cycles can be determined.     

Efficacy of microsurgical epididymal sperm aspiration (MESA) and intracytoplasmic sperm injection (ICSI) in obstructive azoospermia

Purpose: Our purpose was to test whether micromanipulation using subzonal insemination and intracytoplasmic sperm injection could improve the poor fertilization and pregnancy rates obtained when attempting in vitro fertilization in patients with congenital absence of the vas deferens and unreconstructable obstructive azoospermia with microsurgically retrieved epididymal spermatozoa. Results: Conventional in vitro fertilization (group A; 14 cycles), subzonal insemination (group B; 13 cycles), and intracytoplasmic sperm injection (group C; 28 cycles) were carried out in 55 treatment cycles. Fertilization rates for groups A, B, and C were 16.1, 31.4, and 48.6%, respectively (P<0.05). Clinical pregnancy rates for groups A, B, and C were 7.1, 7.7, and 32.1% (P<0.05), respectively. In five cycles, intracytoplasmic sperm injection using epididymal sperm from alloplastic spermatoceles was performed and two clinical pregnancies (40%) were obtained. Conclusions: The combined microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection procedure is highly effective in improving the fertilization and pregnancy rate in congenital absence of the vas deferens and unreconstructable obstructive azoospermia. Furthermore, alloplastic spermatoceles may be useful for repeat sperm aspirations.Presented in part at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, Vienna, Austria, April 3–7, 1995.     

早期补救卵胞浆内单精子显微注射在完全受精失败周期的临床结局分析

目的:评估短时受精联合早期补救卵胞浆内单精子显微注射(R-ICSI)在完全受精失败周期的临床价值。方法:2009年1月～2010年6月我中心试管婴儿助孕治疗709例,其中短时受精完全失败行早期R-ICSI周期82例,卵胞浆内单精子显微注射(IC-SI)周期627例,比较两组正常受精率、异常受精率、优质胚胎率、胚胎种植率、临床妊娠率及流产率。结果:两组正常受精率、优质胚胎率、胚胎种植率、临床妊娠率及流产率无统计学差异,R-ICSI组异常受精率(5.0%)显著高于ICSI组(3.0%)(P<0.05)。结论:短时受精联合早期R-ICSI可及早发现受精失败并及时补救,获得较好的临床结局。     

Effects of failed oocyte activation and sperm protamine deficiency on fertilization post-ICSI

Sperm premature chromosomal condensation (PCC) has been associated with failed fertilization. Previous studies suggest that protamine deficiency or failed oocyte activation may make spermatozoa prone to PCC. However, it is not clear which of these two factors has a more profound effect on fertilization failure. In order to distinguish between these two phenomena, oocytes that failed to fertilize after intracytoplasmic sperm injection (ICSI) were artificially activated and the association between protamine deficiency and PCC was evaluated in the remaining oocytes that failed to fertilize. The results of this study reveal that after artificial activation, fertilization rate post-ICSI increased from 59.95 to 87.7% and PCC spermatozoa appeared to be present in over 50% of the remaining oocytes that failed to fertilize. The percentage of sperm PCC was significantly higher in protamine deficient samples, thus suggesting that after failed oocyte activation, sperm PCC induced by protamine deficiency may be considered as an alternative cause of failed fertilization post-ICSI. Furthermore, the results of this study did not show any correlation between pronuclei size asynchrony and protamine deficiency.     

Failed fertilization: is it predictable?

PURPOSE OF REVIEW: The purpose of this review is to critically examine the ability of screening tests to predict fertilization failure. RECENT FINDINGS: Failed fertilization occurs in 5-10% of in-vitro fertilization cycles and 2-3% of intracytoplasmic sperm injection cycles. Failed fertilization may result from impaired spermatozoa, oocyte deficiencies or defects in the in-vitro sperm/oocyte medium. In the in-vitro fertilization setting most cases are caused by male factor deficiencies, whereas failure of oocyte activation is the most common cause of failed fertilization after intracytoplasmic sperm injection. Although the standard semen analysis has limited ability to predict fertilization failure, strict sperm morphology criteria, sperm-zona binding ratios and zona pellucida induced acrosome reaction tests provide increased capacity to avoid this outcome. The quality of the semen sample on the day of oocyte retrieval and fertilization performance in previous in-vitro fertilization cycles may also guide the appropriate use of intracytoplasmic sperm injection. However, the routine use of the latter technique in cases of non-male factor infertility is contraindicated. SUMMARY: The ever improving techniques highlighted in this analysis offer improved ability to predict failed fertilization.