IVF-ET周期中1原核(PN)及0PN(2Pb)胚胎的染色体组成分析

目的:初步探寻体外受精-胚胎移植(IVF-ET)周期中高发育潜能的1原核(PN)及0PN(2Pb)胚胎的方法。方法:观察787个IVF周期患者的PN及卵裂情况,培养至第6日,观察其囊胚形成情况。取囊胚滋养层细胞活检,采用胚胎植入前遗传学筛查(PGS)技术检测0PN(2Pb)、1PN及2PN囊胚的染色体组成。结果:787个IVF周期共获卵8 352枚,2PN、0PN(2Pb)、1PN胚胎的形成率分别为64.35%、4.93%、3.65%,囊胚形成率分别为45.71%、30.03%、18.18%。PGS结果显示,1PN及0PN(2Pb)囊胚染色体正常的比例明显低于2PN囊胚,差异有统计学意义(P0.05)。0PN(2Pb)囊胚的染色体正常比例较1PN囊胚的略高,差异无统计学意义(P0.05)。此外,发育至第6日的0PN(2Pb)囊胚及1PN囊胚较第5日胚胎的正常染色体比例高。结论:对于本周期无可利用胚胎的患者,建议移植1PN及0PN(2Pb)来源的第6日评分较好的囊胚。     

体外受精中3原核胚胎的发育及可利用价值的探讨

目的:探讨体外受精(IVF)中异常的3原核(PN)胚胎的发育及可利用价值。方法:收集IVF治疗周期中废弃的3PN受精卵204个进行体外培养,观察其发育能力,并与同周期的1 138个2PN受精卵进行比较;采用胚胎植入前遗传学筛查(PGS)技术对由3PN发育成的19枚囊胚进行非整倍体分析。结果:3PN组和2PN组的卵裂率无统计学差异(P0.05);但3PN组囊胚形成率显著低于2PN组[9.6%(19/97)vs 37.9%(204/342),P0.01]。整倍体分析显示,10.5%(2/19)的3PN来源的囊胚为正常二倍体核型。结论:3PN受精卵有继续发育能力;囊胚培养和高通量测序可作为有效筛选异常PN受精卵中正常核型胚胎的一种方法。     

血脂水平与体外受精周期胚胎质量的相关性初步研究

目的:研究不孕症患者血脂水平与体外受精(IVF/ICSI)胚胎质量的相关性。方法:对行IVF/ICSI治疗、符合纳入标准的646例不孕症患者的资料进行回顾性分析,测定、分析血清三酰甘油(TG)、总胆固醇(TC)、高密度脂蛋白(HDL)、非高密度脂蛋白(nonHDL)、低密度脂蛋白(LDL)和脂蛋白(a)[LP(a)]水平与胚胎质量的关系。血脂水平和IVF/ICSI胚胎质量相关性采用Spearman相关分析。结果:患者年龄、体质量指数(BMI)、基础FSH、窦卵泡数、Gn用量及血脂水平临床妊娠组和非临床妊娠组之间均无统计学差异;受精率、卵裂数、优质胚胎数和优质胚胎率临床妊娠组均显著高于未临床妊娠组(P<0.05)。血脂水平和IVF/ICSI胚胎质量相关分析显示:TG和TC与卵子数、受精数及卵裂数均显著负相关,TC、LDL及nonHDL与优质胚胎数呈显著负相关(P<0.05),其余均无统计学意义。结论:血脂水平临床妊娠组与未临床妊娠组间均无统计学差异;IVF/ICSI胚胎质量与TC、LDL及nonHDL呈显著负相关。     

体外受精-胚胎移植中胚胎形态和发育时期与受孕结果的相关性

目的 :观察胚胎形态和发育速度对体外受精 -胚胎移植 ( IVF-ET)治疗结果的影响。方法 :35 4对夫妇进行的 371个 IVF-ET治疗周期 ( ICSI177个周期 ,常规 IVF 194个周期 ) ,根据移植胚胎形态等级将 IVF-ET周期分为 A、B、C、D 4组 ;再根据移植胚胎中有无≥ 4-细胞期胚胎将 IVF-ET周期分为≥ 4-细胞期胚胎组和 <4-细胞期胚胎 2组 ,分析移植胚胎形态等级和卵裂期对胚胎种植率和临床妊娠率的影响。结果 :371个周期共移植 112 5个新鲜胚胎 ,四个形态等级胚胎组之间种植率 ( P=0 .0 12 )和妊娠率 ( P=0 .0 0 6)差别有显著性。两个卵裂期胚胎组之间种植率 ( P=0 .0 0 7)和妊娠率 ( P=0 .0 10 )有显著性差异。 L ogistic回归分析发现胚胎质量 (形态等级 P=0 .0 0 38,卵裂期 P=0 .0 0 45 )是影响临床妊娠结果的因素。结论 :移植胚胎的形态和发育期与胚胎种植率及临床妊娠率密切相关     

受精方式、女方年龄、生育史和取卵数对体外异常受精的影响

目的:探讨人卵子体外受精后单原核(PN)和多原核胚胎形成的影响因素,为提高正常受精率探寻可行的方法。方法:回顾性分析1 004个IVF周期和250个ICSI周期,共计15 364个卵细胞资料,研究胚胎原核形成与体外受精方式、女方年龄、生育史、获卵数的关系。结果:①ICSI周期的1PN率明显高于IVF周期,而3PN和4PN形成率则明显低于IVF(P<0.05)。②当女方年龄>35岁时,1PN、3PN和4PN形成率均显著高于年龄≤35岁者(P<0.05);当女方年龄在28￣35岁时2PN(正常受精率)生成率最高(P<0.05),此年龄段的患者施行ICSI时3PN生成率也最高(P<0.05)。③女方无生育史的1PN形成率上明显高于有生育史患者(P<0.05)。④获取6￣10枚卵组,2PN形成率明显高于1￣5枚卵组和>20个卵组(P<0.05);而对于1PN、3PN、4PN形成率,获11￣15枚卵组明显高于其他获卵数组(P<0.05);而在获卵数>20枚时ICSI组2PN(正常受精率)生成率显著降低。结论:人体外受精正常与否受到受精方式、女方年龄、生育史和取卵数目等因素的影响,因此,在进行辅助生殖技术(ART)时应综合考虑这些因素,有助于提高正常受精率,降低异常受精率。     

体外受精-胚胎移植技术中74例异位妊娠的临床分析

目的:分析体外受精-胚胎移植(in vitro fertilization and embryo tranfer,IVF-ET)助孕技术中发生异位妊娠的影响因素、诊断、治疗方法及预防措施。方法:回顾性分析IVF-ET助孕技术中发生异位妊娠74例患者的临床资料。结果:所有接受IVF-ET的1 585患者中,共发生异位妊娠74例,异位妊娠发生率为4.67%;异位妊娠类型中输卵管妊娠71例,占总异位妊娠的95.95%。新鲜胚胎移植周期组异位妊娠发生率(5.41%)显著性高于冷冻胚胎复苏移植(frozing embryo transfer,FET)周期组(2.35%)(P<0.05)。移植深度距离宫底>1.2 cm时异位妊娠发生率显著性低于移植深度0.8~1.2 cm组。结论:IVF-ET助孕技术中异位妊娠发生率较自然妊娠过程中高;输卵管因素、促排卵药物的应用以及胚胎移植的深度是异位妊娠发生的主要影响因素。     

短时常规体外受精中废弃胚胎染色体非整倍体分析

目的:探讨短时常规体外受精周期中废弃胚胎中染色体非整倍体的情况。方法:收集常规体外受精周期及短时常规体外受精周期1原核(1PN)、2PN和≥3PN的废弃胚胎,运用2轮荧光原位杂交技术对胚胎染色体进行分析。结果:短时常规体外受精组与常规体外受精组废弃胚胎染色体异常率差异无统计学意义。结论:短时常规体外受精技术本身不会增加胚胎染色体异常的几率,是一种安全有效的受精方式。     

体外受精-胚胎移植短方案周期中移植胚胎数与临床妊娠结局的关系

目的:探讨体外受精-胚胎移植(IVF-ET)短方案周期中移植不同胚胎数对于临床妊娠率和多胎发生率的影响。方法:回顾性分析2002.01-2004.10期间进行第一次IVF-ET短方案周期治疗、年龄<35岁的患者1463例,将2002.01-2003.09间移植2个胚胎者为A组(n=84)、移植3个胚胎者为B组(n=716);2003.10-2004.10期间移植2个胚胎者为C组(n=663)。分析和比较3组的可移植胚胎数、胚胎种植率、临床妊娠率和多胎发生率等。结果:B组的可移植胚胎数(7.8±3.7)显著高于移植2个胚胎的A组(4.6±4.7)和C组(6.9±3.9),P均<0.05,胚胎种植率各组间无显著性差异(A:20.62%,B:14.88%,C:21.66%),P均>0.05。B组的临床妊娠率(42.96%)显著高于A组(25.93%)和C组(39.06%),P均<0.05;各组间的单胎妊娠率无显著性差异(A:27.19%,B:20.99%,C:29.42%,P均>0.05);A组(4.94%)和C组(10.06%)的多胎妊娠率均比B组(15.77%)显著降低,P<0.05。结论:IVF-ET短方案周期中,减少移植胚胎数可能会降低临床妊娠率,但移植2枚胚胎能够有效降低多胎妊娠的发生。     

父源性染色体异常胚胎植入前遗传学诊断的最新研究进展

染色体异常是导致男性不育的一个重要原因。不同类型染色体异常的生殖细胞进行减数分裂时形成不同类型的配子,且各种异常配子所比例的实际值与理论值不相符。另外,正常胚胎率与正常配子率也不一致。本文对染色体结构异常(相互易位、罗伯逊易位、倒位、染色体复杂性重组)和数目异常(性染色体数目异常、常染色体数目异常)男性的正常精子率以及正常胚胎率进行综述。     

胚胎分泌的可溶性人类白细胞抗原G(sHLA-G)与IVF临床妊娠率的相关性研究

目的:探讨可溶性人类白细胞抗原G(sHLA-G)与IVF临床妊娠率的关系。方法:收集IVF-ET患者体外单个培养胚胎的培养液,通过ELISA法检测培养液中sHLA-G的浓度,并计算相应移植胚胎的妊娠率。结果:在41名患者的84份胚胎培养液中,sHLA-G阳性率52%(44/84),阳性培养液中sHLA-G浓度为3.7￣16.6ng/ml;移植至少一个sHLA-G阳性胚胎的妊娠率为69%,移植胚胎全部为sHLA-G阴性的妊娠率为18%,两者相比有统计学差异(P<0.05)。结论:以sHLA-G阳性表达水平结合传统胚胎形态学评估方法,可以作为辅助选择移植胚胎的临床指标,预测IVF妊娠成功率。     

Human Embryos Derived from In Vitro and In Vivo Matured Oocytes: Analysis for Chromosomal Abnormalities and Nuclear Morphology

Purpose: To determine whether embryos resulting fromoocytes matured in vitro have a higher incidence of nuclearand/or genetic abnormalities compared to embryos resultingfrom oocytes matured in vivo. Methods: Fluorescence in situ hybridization analysis forchromosomes X, Y, and 18 was used to compare the ratesof aneuploidy, mosaicism, and nuclear abnormalities inembryos derived from oocytes that were prophase I ataspiration (immature group) to that observed in embryos resultingfrom oocytes that were metaphase I or II at aspiration(mature group). Results: Based on nuclear morphology, significantly moreembryos in the mature group (23percnt;) were classified as normalcompared to embryos in the immature group (3percnt;). Nodifference was found in the rate of aneuploidy or in the incidenceof mosaicism involving these chromosomes. Conclusions: These findings suggest that few embryosderived from prophase I oocytes collected following ovarianstimulation are morphologically normal.     

人类体外受精培养液内毒素水平的评价

目的:检测人类体外受精使用的商品性培养液的内毒素水平,评价人精子存活试验和小鼠2-细胞发育试验测试内毒素的差异。方法:36批次的商品性体外受精培养液,使用前(A组,n=36),及使用后的其中25份样品(B组,n=25)采用鲎试验法检测内毒素水平。另对比人精子存活试验和2-细胞胚胎发育试验的敏感性。结果:A组样品无内毒素阳性检出,B组有2份样品检出内毒素。A组和B组样品24h精子活动率改变与对照组比无显著性差异(P>0.05),但A组中有3份样品抑制小鼠2-细胞胚胎的发育。结论:体外培养环境和实验操作须防止内毒素污染。虽然商品性培养液未检出内毒素,仍须对其作严格的质量控制。人精子存活试验测试培养液质量和低内毒素水平的敏感性低于小鼠2-细胞发育试验。     

Ethical Dimensions of the Number of Embryos to be Transferred in In Vitro Fertilization

Purpose: We propose an ethically justified policy for the number of embryos to transfer in an in vitro fertilization (IVF), by considering four factors: medical outcomes, patient's preferences, costs, and market forces of providers.Methods: We develop an ethical framework that incorporates three ethical principles: beneficence, respect for autonomy, and justice; and three professional virtues: integrity, compassion, and self-sacrifice.Results: This ethical framework calls for an informed consent process for IVF that provides: information about medical outcomes; information about the risks of multiple gestation; the opportunity to weigh the goal of pregnancy and live birth against the medical and moral risks of multiple gestation; evidence-based recommendations; protection of the woman from potentially coercive influences; and discussion of living with infertility and going to better centers.Conclusion: The number of embryos to be transferred in IVF should mainly be a function of the pregnant women's informed decision. Limiting the number of transferred embryos to two in all cases is not ethically justified at this time.     

Effects of Reducing Insemination Time in Human In Vitro Fertilization and Embryo Development by Using Sibling Oocytes

Purpose: Recent studies showed a beneficial effect of reducing the time of sperm–oocyte interaction on fertilization, division, and implantation rates of the oocytes obtained from randomized patients. In the present study, the effects of reduced insemination time on fertilization and embryo development were evaluated by using sibling oocytes from the same patient. Methods: A total of 464 oocytes from 36 patients was randomly allocated to be inseminated for either 1 hr (reduced) or 18 hr (regular). Results: Fertilization rates were not significantly different between reduced (135/229; 59%) and regular (150/235; 64%) groups. Cleavage rates and embryo quality were similar in both groups. A total of 135 embryos (73 from the reduced and 62 from the regular group) was transferred to 36 patients. Thirty-four embryos implanted in 18 patients (25.2% implantation and 50.0% pregnancy rates). Conclusions: Fertilization, cleavage, and embryo development from 1-hr insemination is comparable, not superior, to those from an 18-hr insemination time, which is commonly used in in vitro fertilization programs. These data suggest that reduced insemination time can be used during in vitro fertilization to avoid unnecessarily longer exposure to spermatozoa.     

Culture with Matrigel Inhibits Development of Mouse Zygotes

Purpose: It was reported that Matrigel improved hatching of mouse blastocysts produced in vitro from F ₁ hybrid-derived zygotes. We investigated whether Matrigel would be similarly beneficial with outbred strain-derived embryos, which exhibit a two-cell block similar to the developmental blocks of other species. Methods: Mouse embryo development was assessed with or without Matrigel in KSOM medium, which supports the development of blocking strain zygotes in vitro, and in human tubal fluid (HTF) medium, which normally does not but which is used for human IVF. Results: Matrigel severely inhibited the development of zygotes to blastocysts in KSOM and did not improve culture in HTF. There was no effect on development from the two-cell stage. We were not able to replicate the previous finding of Matrigel's beneficial effect on hatching of F ₁ -derived zygotes. Conclusions: Matrigel may be a deleterious addition to embryo culture or coculture systems.     

体外受精后多原核受精卵的移植价值

目的:对体外受精周期中产生多原核受精卵运用荧光原位杂交技术(FISH)进行非整倍体率检测,分析其移植价值。方法:应用FITC、Texas red标记的X/Y双色染色体着丝粒部位探针对体外受精(IVF)和单精子显微注射(ICSI)周期中的三原核受精卵(3PN)进行荧光原位杂交和分析。结果:分析有杂交信号的3PN卵子68个,IVF后3PN的三倍体率为92.31%,二倍体率为3.85%,单倍体率为3.85%;共存在3种三倍体核型:XXX、XXY、XYY。ICSI后3PN 的三倍体率为70.83%,二倍体率为25%,单倍体率为4.17%;共存在2种三倍体核型:XXX、XXY。结论:IVF后3PN卵子中多余的原核多为精子来源,没有移植价值;ICSI后3PN中多余的原核非精子来源,ICSI后3PN中二倍体率与IVF相比显著增高,在患者ICSI后无2PN卵子时可考虑行植入前遗传学诊断。     

Fertilization, Embryo Quality, and Cryosurvival in In Vitro Fertilization and Intracytoplasmic Sperm Injection Cycles

Purpose: Our purpose was to investigate the influence of semen quality on fertilization, embryo morphology, cleavage, and cryosurvival in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI)programs. Methods: A retrospective analysis of 513 couples undergoing IVF and 255 couples undergoing ICSI was done. Results: Semen quality influenced fertilization in IVF and abnormal fertilization in IVF and ICSI, but no effects on the development, morphology, implantation capacity, or cryosurvival of embryos were found. Fertilization, embryo quality, and cryosurvival rates were similar after IVF and ICSI. The fertilization rate of mature oocytes in IVF was lower when cytoplasmic immaturity in the oocyte population was frequent. The speed of development of embryos was 2 hr faster after ICSI than after IVF. Two-cell–stage embryos survived best after cryopreservation with propanediol and sucrose on day 2. Conclusions: After fertilization, semen parameters had no effect on the quality or cryosurvival of embryos in either IVF or ICSI.     

Incidence of Chromosomal Abnormalities from a Morphologically Normal Cohort of Embryos in Poor-Prognosis Patients

Purpose: Preimplantation genetic diagnosis of aneuploidy was performed on the embryos yielded by 70 poor-prognosis patients, with the aim of transferring those with a normal chromosomal complement, thus possibly increasing the chances of pregnancy. Methods: Multicolor fluorescence in situ hybridization (FISH) was applied for the simultaneous detection of chromosomes X, Y, 13, 16, 18, and 21. Inclusion criteria were (1) a maternal age of 36 years or older (n = 33), (2) three or more previous in vitro fertilization cycles (n = 20), and (3) an altered karyotype (n = 17). Results: A total of 412 embryos underwent FISH, resulting in 234 (57%) that were chromosomally abnormal. Euploid embryos were available for transfer in 59 patients, generating 19 pregnancies (32%), with an implantation rate of 19.9%. Conclusions: High rates of chromosomally abnormal embryos in poor-prognosis patients can determine repeated in vitro fertilization failures when embryo selection is performed on the basis of morphological criteria alone. Hence, the FISH analysis could represent the prevailing approach for the identification of embryos possessing full potential for developing to term.     

Smoking and In Vitro Fertilization: A Meta-Analysis

Purpose: Including our own data and seven relevant publications from the literature, this meta-analysis aimed to establish the influence of the status of female smokers on the clinical pregnancy rate after the first attempt at in vitro fertilization–embryo transfer (IVF-ET). Methods: We started to collect information about our own patient's smoking habits in January 1996 to evaluate prospectively the influence on IVF success. Information regarding 799 patients (607 nonsmokers and 192 smokers) was obtained. Additionally we retrieved seven relevant publications from the unabridged MEDLINE1982–1996. They were recalculated to first cycles, because from a biometrical point of view, only the results of the first IVF cycle are really well comparable, as the number of necessary IVF cycles always correlates with the number of pregnancies. These data were compared in the nonparametric sign test according to Dixon–Mood and calculated as relative success ratios, defined as the quotient of the probability of IVF-ET success of nonsmokers divided by that of smokers for each publication. Results: This meta-analysis, based on a total of 2314 first IVF-ET treatments, showed that almost twice as many IVF-ET cycles were needed for smokers as for nonsmokers to become pregnant. For the success quotient, we obtained a ratio of 1.79, with an associated 95% confidence interval of from 1.24 to 2.59. The total analysis shows, with a P value of less than 0.01, significantly higher pregnancy rates (21%) in nonsmokers compared to smokers (14%). Conclusions: Based on the analyzed data there exists—from a statistical point of view—a significant negative effect on the chances of success for smokers to become pregnant compared to nonsmokers.