体外受精失败的次级卵母细胞单精子显微注射后的效果观察

目的：探讨采用精子卵胞浆内注射（ICSI）对于常规体外受精和胚胎移植（IVF－ET）受精失败的次级卵母细胞实施补救受精的效果。方法：对在常规IVF－ET中没有得到受精卵的病例的次级卵母细胞实施ICSI，观察受精、卵裂、胚胎种植和受孕情况。结果：2000年1月～2000年12月在我院生殖中心接受常规IVF治疗的夫妇中，12例在授精后12～20 h卵子全部不受精，在取卵后24 h实施ICSI。12例患者有卵111个，对其中的69个次级卵母细胞实施ICSI， ICSI 20 h后检查，56个受精，受精率81．2％，形成胚胎50个，卵裂率89．3％，移植11个周期共38胚胎，临床妊娠1例（双胎）。胚胎着床率为5．3％。结论：在常规IVF无受精的情况下，应当立即实施ICSI以补救。但在此情况下，胚胎的着床率低下。

减少卵子老化对于补救性卵母细胞单精子显微注射结局的影响

Objective: To evaluate the results of a novel protocol that allows to rescue IVF unfertilized oocytes by intracytoplasmic sperm injection (ICSI).Design: Prospective clinical trial.Setting: Private reproductive medical center.Patient(s): Thirty patients undergoing IVF.Intervention(s): Controlled ovarian stimulation (COS), conventional IVF, rescue ICSI, embryo culture, and embryo transfer.Main Outcome Measure(s): Identification of unfertilized IVF oocytes 6 hours after insemination and fertilization, and developmental rates of those oocytes after rescue microinjection, as well implantation and pregnancy rates (PR).Result(s): All oocytes (392) from 30 patients were inseminated with standard IVF 3 hours after ovum pick- up.Polar body (PB) status was checked at decumulation and rechecked 3 hours later.Eighty- two oocytes were fertilized after IVF alone and 184 nonactivated oocytes (failed fertilization) were rescue microinjected and 166 of them fertilized (20 patients).Cleavage stage on day 2 was significantly more advanced and embryo grade was higher after standard IVF fertilization than after rescue ICSI.Eight of the 30 embryos transferred were implanted in the IVF- only patients (27% )- and 8 of 68 embryos in the rescue ICSI patients (12% ).Conclusion(s): Rescue ICSI of unfertilized IVF oocytes 6 hours after insemination (9 hours after egg retrieval) can provide normal fertilization, embryo development, and pregnancy; however, corresponding outcome parameters tend to be impaired in comparison to the standard IVF fertilization results.

在体外受精和卵母细胞单精子显微注射结局中由精子染色质散布试验测得的精子脱氧核糖核酸碎片水平的价值

Objective: To determine the prognostic value of sperm DNA fragmentation levels, as measured by the sperm chromatin dispersion (SCD) test, in predicting IVF and ICSI outcome. Design: Double- blind prospective study. Setting: University- affiliated private IVF setting. Patient(s): A total of 85 couples undergoing infertility treatment with IVF/ICSI. Intervention( s): Analysis of DNA fragmentation by the SCD test in 170 aliquots obtained from the ejaculate and from the processed semen used for assisted reproductive technologies (ART). Main Outcome Measure(s): Percentage of spermatozoa with fragmented DNA was statistically correlated with embryo quality and reproductive success. Result(s): Fertilization rate was inversely correlated with DNA fragmentation (r =- 0.245 P=.045). Higher DNA fragmentation rate gave an increased proportion of zygotes showing asynchrony between the nucleolar precursor bodies of zygote pronuclei (73.8% vs. 28.8% P < .001). In addition, the slower embryo development and worst morphology on day 6 was correlated with higher sperm DNA fragmentation (47.7% vs. 29.4% P=.044). We also observed a negative correlation between DNA fragmentation and the implantation rate (r =- 0.250 P=.042). However, SCD test values were not statistically different in cycles that resulted in a pregnancy compared with those that did not (33.2 vs. 28.2 and 32.4 vs. 34.7). Conclusion(s): This is the first report that describes a correlation between sperm DNA integrity, asmeasured by the SCD test, and fertilization rate, embryo quality, and implantation rate in IVF/ICSI. The degree of DNA fragmentation was inversely correlatedwith fertilization rate, synchrony of the nucleolar precursor bodies’ pattern in pronuclei, embryo ability to achieve blastocyst stage, and embryo morphological quality. Because SCD test values were correlated with embryo quality and blastocyst rate, the lack of correlation between sperm DNA fragmentation and pregnancy outcome in IVF might be due to embryo selection before transfer. The ability of the SCD test to predict the blastocyst rate after IVF/ICSI warrants further study.

行卵母细胞单精子显微注射且年龄≥40岁的患者使用促性腺激素释放激素类似物进行控制性超促排卵治疗短期或长期方案的比较

To determine whether the short or long protocol for controlled ovarian hyperstimulation works better in older patients undergoing IVF. Design: Controlled, randomized study. Setting: A single private IVF center. Patient(s): Two hundred twenty infertile women aged ≥ 40 years undergoing IVF.Intervention(s): At their first IVF cycle, the women were randomized into two stud groups according to a computer-generated number sequence: 110 patients were treated with a long protocol, and the other 110 were treated with a short protocol for controlled ovarian hyperstimulation. Main Outcome Measure(s): Days of stimulation, E2 level at the day of hCG administration, amount of FSH administered, number of oocytes collected, number of embryos obtained, pregnancy rate, implantation rate. Result(s): Patients treated with a long protocol showed a significantly higher number of oocytes retrieved, a higher number of embryos obtained, and a higher pregnancy rate, both for cycle and transfer, compared with the short-protocol patients. The other parameters evaluated did not show any statistically significant differences. Conclusion(s): Our study showed that the long protocol performed better than the short protocol in older women. Our findings demonstrated that flare-up in older women might be detrimental.

辅助孵化对卵母细胞单精子显微注射结局的影响:一项前瞻性随机临床试验及妊娠追踪观察

Objective: To evaluate the overall effect of assisted hatching (AH) on the implantation, pregnancy, and live birth rates in women undergoing intracytoplasmic sperm injection (ICSI)- cycles; and to determine the effect of AH on the cytogenetic outcome (chromosomal constitution) of pregnancy.Design: Prospective, randomized study.Setting: Academic research environment.Patient(s): A total of 172 couples were enrolled in the study.Intervention(s): Assisted hatching was carried out on day- 3 ICSI embryos.Main Outcome Measure(s): Implantation, clinical pregnancy, and live birth rates; cytogenetic analysis of abortuses and umbilical cord blood samples from newborns.Result(s): Biochemical, clinical, and ongoing pregnancy rates were not significantly different between the AH and control groups.The implantation rate was higher in the AH group than in the control group (16% vs.8% ), especially in women aged ≥ 35 years.Postnatal umbilical cord blood samples were collected and cytogenetically analyzed from 39 live births (20 from the AH group, 19 from the control group).Two abnormal karyotypes were found (one AH, one control).There were seven spontaneous losses during the study interval.Six of the abortuses underwent cytogenetic study (five AH, one control), and four were found to have an abnormal karyotype (three AH, one control). Conclusion: We found that AH improves implantation rates of ICSI cycles and seems to be most effective in women aged ≥ 35 years.A larger sample size is needed to determine whether AH improves the take- home- baby rate.Assisted hatching did not affect the rate of chromosomal abnormalities in live births in this study.

针灸对于体外受精和卵母细胞单精子显微注射结局的影响:一项前瞻性的随机对照临床研究

Objective: To determine the effect of luteal-phase acupuncture on the outcome of IVF/intracytoplasmic sperm injection (ICSI). Design: Randomized, prospective, controlled clinical study. Setting: University IVF center. Patient(s): Two hundred twenty-five infertile patients undergoing IVF/ICSI. Intervention( s): In group I, 116 patients received luteal-phase acupuncture according to the principles of traditional Chinese medicine. In group II, 109 patients received placebo acupuncture. Main Outcome Measure(s): Clinical and ongoing pregnancy rates. Result(s): In group I, the clinical pregnancy rate and ongoing pregnancy rate (33.6% and 28.4% , respectively) were significantly higher than in group II (15.6% and 13.8% ). Conclusion(s): Luteal-phase acupuncture has a positive effect on the outcome of 5 IVF/ICSI. .

卵母细胞单精子显微注射可能导致男胎Y染色体微缺失的垂直传播、扩展和新生

Y-chromosome microdeletion in male fetuses conceived by intracytoplasmic sperm injection (ICSI) was screened by polymerase chain reaction for sequence-tagged sites in azoospermia factor (AZF)-b and AZFc regions. Treatment with ICSI may lead to vertical transmission, expansion, and de novo Y-chromosome microdeletion in male fetuses.

卵母细胞单精子显微注射后卵母细胞退化:多因素分析评估其作为实验室或临床标记的重要性

Objective:Oocyte degeneration has historically been associated with the intracytoplasmic sperm injection(ICSI) technique. We sought to determine whether oocyte degeneration rates were associated with the technician performing the procedure,the baseline characteristics of the patient,and/or ovarian stimulation variables. We also evaluated whether the degeneration rate could serve as a surrogate marker for implantation potential. Design:Cohort study. Setting:Academic medical center. Patient(s):Couples undergoing ICSI. Intervention(s):Six thousand six hundred fifty-three injected oocytes were analyzed to determine whether the degeneration rate was technician dependent. Two hundred thirty first-entry down-regulated cycles were examined to identify predictors associated with oocyte degeneration. Multivariate analyses were performed using generalized linear model routines. Main Outcome Measure(s):Oocyte degeneration rates and implantation rates. Result(s):Neither the ICSI technician nor the stripping technician was associated with the oocyte degeneration rate. However,the day 3 FSH,number of mature oocytes retrieved,and E2 levels on the day of hCG were significant independent predictors of degeneration rate. Physician-adjustable ovarian stimulation variables were not associated with the degeneration rate. The degeneration rate did not appear to be associated with the implantation rate. Conclusion(s):These data suggest that oocyte degeneration is not technician or physician dependent. Degeneration is likely a function of the inherent oocyte quality in women who underwent ovarian stimulation. However,the remaining cohort of retrieved oocytes appears to be unaffected by virtue of an uncompromised implantation rate.

体外受精与卵母细胞单精子显微注射周期中不同受精过程对胚泡形成的影响

Objective: To compare the quality of early cleaving embryos and blastocysts ob tained by IVF or intracytoplasmic sperm injection (ICSI). Design: Retrospective study. Setting: Tertiary infertility center. Patient(s): Sibling oocytes of 104 patients in 104 IVF vs ICSI cycles. Intervention(s): Cumulus oocyte complexes (n = 1,358) were randomly subjected to ICSI or IVF. Main Outcome Measure(s): Embry o development and blastocyst formation rate. The blastocyst quality and cycle ef ficiency were also evaluated. Result(s): Early embryo cleavage was higher after ICSI (37.1%) compared to IVF (14.1%). The percentage of < 4-cell embryos on d ay 2 and < 8-cell embryos on day 3 was similar for both procedures. The overall blastocyst formation was not different between ICSI (50.2%) and IVF (54.8%), neither was the percentage of good-quality blastocysts (31.3%for ICSI and 36.0 %for IVF). The total cycle efficiency (percentage of embryos transferred and fr ozen per two pronuclei [2PN]) was comparable for the two techniques (51.7%for I CSI and 57.4%for IVF). Conclusion(s): No differences were found on sibling oocy tes in the embryo development and blastocyst formation, irrespective of the fert ilization procedure. Earlier suggestions that the ICSI technique may result in i mpaired blastocyst development were not confirmed in this study.

卵母细胞单精子显微注射后娩出脐部联体双胎

To describe a first case of parasitic twin achieved after intracytoplasmic sperm injection (ICSI). Case report. Perinatal center at a Japanese university hospital. A 30-year-old pregnant Japanese woman with a history of secondary infertility achieved after ICSI was diagnosed with an omphalopagus parasitic twin pregnancy by prenatal ultrasound at 28 weeks of gestation. A female infant was delivered by scheduled cesarean section at 30 weeks of gestation. The cesarean section was performed for dystocia. Brief review of parasitic twin and malformations in fetuses achieved after ICSI. Resuscitation was not done because of the poor prognosis for both fetuses. Molecular analysis with informative genetic markers is consistent with monozygotic pregnancy. A careful ultrasound examination is indicated to detect additional anomalous findings in twin fetuses achieved after ICSI.

多次经直肠电射精对精子参数及卵母细胞单精子显微注射结果的影响

Objective To compare semen parameters and intracytoplasmic sperm inje ction (IC SI) outcome in spinal cord-injured subjects who underwent single (group 1) or m ultiple (group 2) electroejaculations before ICSI. Design Prospective, randomize d, controlled study. Setting Department of gynecology, obstetrics, and pediatric science in a reproductive medicine unit at a major Italian university. Patient( s) Thirty-four healthy women with a male partner with SCI who were seeking assi sted reproduction services. Intervention(s) Transrectal electroejaculation, cont rolled ovarian hyperstimulation, and ICSI. Main outcome measure(s) Sperm concent ration, morphology, and motility and fertilization and pregnancy rates after ICS I. Result(s) Sperm was successfully retrieved in 94.1%of cases. In male subject s who underwent multiple electroejaculations, statistically significant improvem ents in sperm concentration and total sperm motility rate were observed. The ove rall fertilization rate was 63.6%. The number of oocytes retrieved and injected was comparable between the two groups. A total of nine clinical pregnancies wer e achieved. The pregnancy rate was statistically significantly higher in group 2 (n = 6/16; 37.5%) than in group 1 (n = 3/16; 18.75%). Conclusion(s) These dat a suggest that multiple electroejaculation has a positive effect on semen parame ters and ICSI outcome.

睾丸活检组织学结果的变异与卵母细胞单精子显微注射治疗男性不孕症的相关性

To determine whether center to center discrepancies in the ability to locate sperm in infertile testes with abnormal histology stems in part from inconsistencies in pathologists’ readings of testis biopsies. Design: Prospective cohort study. Setting: Academic male infertility practice. Patient(s): Consecutive series of azoospermic men referred with testis biopsy slides between 1998 and 2003. Intervention(s): Testis biopsy histologies on azoospermic patients referred for infertility care were re-reviewed by a single pathologist blinded to the original reading. Subsequent infertility care was guided by the findings from the second histologic reading. Main Outcome Measure(s): Agreement between the outside and in-house review of testis biopsy readings was assessed with the kappa statistic. Pregnancy outcomes that resulted from clinical decisions informed by the second histologic readings were also assessed. Result(s): Among 113 histologic specimens, re-review was complicated by fixation artifacts in 18 cases (16% ) and insufficient biopsy sample size in 13 cases (12% ). The kappa score for interobserver agreement in readings was 0.43 (95% CI 0.32- 0.54). Mixed histology patterns in particular were underappreciated by outside pathologists (13% of cases on original reading, 36% of cases on review). In 27% of all cases, the differences in biopsy readings had a significant impact on clinical management. Conclusion(s): A correlation between independent testis histology readings in azoospermic men demonstrates frequent inconsistencies. These differences contribute to inaccurate phenotyping of male infertility and can significantly impact the direction of infertility care. These findings highlight the need for a standardized approach to testis histologic review.

轻度的少畸弱精症夫妇与精子正常的夫妇行传统体外受精与卵母细胞单精子显微注射的比较

Use of intracytoplasmic sperm injection(ICSI )in couples with mild oligoteratoasthenozoosp ermia decreases the com-plete fertilization failure rate an d may also reduce the em-bryo cleavage rate.ICSI does not ben efit couples with normal sperm.

补救卵母细胞单精子显微注射和植入前遗传学诊断联合获得妊娠

Objective: To report the birth of a baby from the transfer of one embryo after rescue intracytoplasmic sperm injection (ICSI) and preimplantation genetic diagnosis (PGD). Design: Case report. Setting: IVF center. Patient(s): A 42- year-old G2P0020 woman with unexplained infertility. Intervention(s): Rescue ICSI followed by PGD. Main Outcome Measure(s): Pregnancy. Result(s): Rescue ICSI was performed on seven unfertilized oocytes, which resulted in three embryos. The PGD analysis revealed one normal embryo, which was transferred and resulted in pregnancy and delivery. Conclusion(s): Rescue ICSI in combination with PGD can result in a successful pregnancy.

既往卵母细胞单精子显微注射(ICSI)不良结局和有限的第2次有丝分裂中期卵细胞的患者应用激光辅助的ICSI

In this study, laser-assisted intracytoplasmic sperm injection (ICSI) improved the fertilization rate and the embryo quality in patients with a history of poor ICSI outcome and with limited metaphase II oocytes. This technique is less traumatic to the oocytes during the procedure, and the use of the technique may be expanded.

阴道补充孕酮可提高卵母细胞单精子显微注射的种植率与妊娠率而对体外受精无影响:一项回顾性研究

Objective: To compare the effectiveness of vaginal progesterone supplementatio n with intramuscular supplementation in assisted reproduction cycles. Design: Re trospective study. Setting: Physiopathology of Human Reproduction Unit, Pordenon e, Italy, from July 2000 to June 2004. Patient(s): Three hundred and eight-five intracytoplasmic sperm injection (ICSI)-procedures (188 with vaginal gel and 1 97 with intramuscular progesterone) and 373 in vitro fertilization (IVF) cycles (227 with vaginal and 146 with intramuscular progesterone). Intervention(s): Pro gesterone luteal supplementation: vaginal gel (Crinone 8%90 mg/day) or intramus cular (Prontogest 50 mg/day). Main Outcome Measure(s): Implantation rates, and t otal and clinical pregnancy rates. Result(s): Higher rates of implantation and t otal and clinical pregnancies were observed in the vaginal supplemented ICSI sub group than in the intramuscular one. This difference was observed for all transf ers (13.3%vs. 8.8%, 39.8%vs. 23.3%, and 28.7%vs. 18.6%)and for ultrasound -guided transfers (17.2%vs. 9.3%, 49%vs. 27%, and 36.9%vs. 21.1%, respect ively). Conclusion(s): The vaginal route of luteal supplementation may be better than the intramuscular one, yielding higher implantation rates as well as total and clinical pregnancy rates in ICSI cycles but not in classic IVF treatments.

行体外受精和卵母细胞单精子显微注射后多原核孕卵形成患者在单胚胎移植后分娩正常婴儿

To report an unusual case of idiopathic zygotic polypronuclei. Case report. Ma jor urban infertility referral center. A 34-year-old woman with unexplained in fertility. The patient underwent two cycles of controlled hyperstimulation and o ocyte retrieval followed by in vitro insemination and intracytoplasmic sperm inj ection (ICSI). Pregnancy and delivery of a normal infant following transfer of a single preembryo from an ICSI cycle in which only one zygote showed a normal pr onuclear number (2PN) and 12 zygotes appeared polypronucleated (< 3PN). On the f irst IVF cycle, 23 oocytes were retrieved and inseminated with 240×103 motile s perm/mL, after which two zygotes showed a normal pronuclear number and 20 zygote s appeared polyploid with three to seven pronuclei. Transfer of two poorquality day-3 preembryos following assisted hatching did not achieve pregnancy. On the subsequent ICSI cycle, 33 oocytes were retrieved, and 17 mature oocytes were sub jected to ICSI, after which only one zygote showed 2PN and 12 zygotes appeared polyploid with three to eight pronuclei. Th e normally fertilized zygote developed into a poor-quality, day-3 embryo and w as subjected to assisted hatching. Transfer of this preembryo resulted in an une ventful pregnancy and birth of a normal infant. Mechanisms other than polyspermi a may result in polypronuclear development in some patients.

严重与轻度少精症患者精液中非整倍体发生率的比较及其对卵母细胞单精子显微注射后妊娠结局的影响

Objective: To determine the incidence of disomy and diploidy for chromosomes 18, X, and Y in the sperm samples of severe oligozoospermic (< 5 ×106 spermatozoa/mL) and oligozoospermic (5-20 ×106 spermatozoa/mL) men undergoing intracytoplasmic sperm injection (ICSI) and to evaluate the influence of sperm aneuploidy on pregnancy outcome. Design: Prospective study. Setting: Infertility clinic and genetic laboratory. Patient(s): Fifteen patients with severe oligozoospermia, 15 patients with oligozoospermia, and 10 normal fertile donors. Intervention(s): Fluorescence in-situ hybridization (FISH) performed on sperm samples. Main Outcome Measure(s): The frequency of disomy and diploidy for chromosomes 18, X, and Y was analyzed using FISH, and the clinical outcome after ICSI was correlated. Result(s): Significantly greater frequencies of XY, YY disomy and diploidy were observed in severe oligozoospermic men compared with oligozoospermic and normozoospermic men. Although the fertilization rate was similar, the pregnancy rate was higher in the group with oligozoospermia versus severe oligozoospermia. Conclusion(s): This study demonstrated the presence of an elevated sperm aneuploidy rate in patients with low semen quality. Additionally, the data show a negative influence of sperm chromosome abnormalities on ICSI outcome.

卵母细胞单精子显微注射过程中使用含有羟乙基哌嗪乙磺酸(HEPES)的缓冲液对体外受精的结局有害

Objective: This study was conducted to determine whether N-hydroxyethylpiperazine-N-ethanesulfonate (HEPES)-buff-ered medium used for the microinjection of sperm into oocytes may be detrimental for the embryo. Design: Controlled randomized study. Setting: Private IVF center. Patient(s): Women (n = 708) undergoing ICSI. Intervention(s): The women were randomized into two study groups: 2,204 oocytes from 357 women were treated using a medium buffered with bicarbonate without HEPES during the ICSI procedure, and 2,168 oocytes from 351 women were treated using a medium buffered with HEPES during the ICSI procedure. Main OutcomeMeasure(s): Fertilization rate, degeneration rate, triploid rate, cleavage rate, embryo quality, pregnancy rate, implantation rate, and abortion rate. Result(s): Oocytes treated with a HEPES-buffered medium showed a statistically significant higher rate of triploid and degenerated oocytes after fertilization with ICSI compared with oocytes treated with a medium without HEPES. The embryos obtained from oocytes microinjected with a HEPES buffered medium showed a statistically significant higher rate of highly fragmented embryos compared with the controls. Pregnancy rate and implantation rate were statistically significantly lower in the patient group with oocytes treated with the HEPES buffered medium. The other parameters evaluated did not show any statistically significant differences. Concluxion(s): Our study showed that the use of media buffered with HEPES, during the microinjection of sperm into the oocytes, is detrimental for IVF outcome and should be avoided.

通过同卵共享模型研究经体外受精和卵母细胞单精子显微注射后男性因素对受孕及胚胎发育的影响

Objective To investigate whether sperm from different males can influ ence fert ilization and embryo development. Design To use an egg-sharing model, in which the eggs from one woman are shared between herself and a recipient, and differen t spermatozoa are used to fertilize the eggs. Setting Assisted Conception Unit, Birmingham Womens Hospital, Edgbaston, United Kingdom. Patient(s) Infertile wo men undergoing egg sharing. Intervention(s) In vitro fertilization (IVF). Main o utcome measure(s) Fertilization rates and the mean day 2 or 3 embryo score (cell number X grade) were examined for eggsharing pairs. A comparison was also made for pairs in which intracytoplasmic sperm injection (ICSI) and IVF was used as t he insemination method. A paired samples t-test was used to compare the sharer and recipient results. Result(s) Pregnancy rates did not differ between sharer a nd recipient couples. Interestingly, when comparing fertilization, there was a s ignificant difference (P <.05) in favor of IVF over ICSI.When comparing embryo d evelopment between egg-sharing pairs, we found that approximately 30%of patien ts showed a difference in mean embryo score of ≥5 in all embryo development and 14%in the quality of embryos available for transfer. Conclusion(s)We showed th at the egg-sharing model is a successful alternative for the treatment of women who required donated eggs. More important, the egg-sharing model shows that, i n a certain percentage of couples, differences in early embryo development are p aternally influenced.