促排卵方案对染色体易位携带者胚胎植入前检测结局的影响

目的探讨促性腺激素释放激素(Gn RH)激动剂长方案与拮抗剂方案对染色体易位携带者胚胎植入前检测(PGT)结局的影响。方法回顾性分析2015年1月至2017年12月于中国医科大学附属盛京医院行胚胎植入前检测助孕的226例染色体易位携带夫妇的临床资料,根据促排卵方案分为长方案组(174例)和拮抗剂组(52例),对比两组间Gn用量、Gn天数、胚胎质量、正常和平衡胚胎数、每移植周期临床妊娠率、流产率、持续妊娠率等差异。结果长方案组和拮抗剂组的获卵数、MⅡ卵子数、正常受精率、卵裂率、囊胚形成率、优质囊胚数、活检囊胚数、正常和平衡胚胎数、每移植周期临床妊娠率、流产率、持续妊娠率和累积持续妊娠率差异均无统计学意义(P0.05)。而拮抗剂组Gn用量(2100 U vs. 2400 U)和天数(9 d vs. 10 d)显著少于长方案组(P0.05)。结论在胚胎植入前检测助孕过程中,长方案与拮抗剂方案的胚胎质量和妊娠率无明显差异,但拮抗剂方案在Gn用量和天数上占有优势。     

胚胎源性PAF水平联合胚胎质量对体外受精-胚胎移植结局的预测价值

目的:探讨接受IVF/ICSI-ET治疗的不孕妇女Day3胚胎培养液中PAF水平对IVF结局的影响。方法:选取接受IVF/ICSI-ET助孕且当月移植新鲜胚胎的85例患者,按移植胚胎质量分为优质胚胎组(64例)和非优质胚胎组(21例)。于Day3收集移植胚胎的培养液并测定PAF含量。结果:移植优质胚胎组患者的植入率、临床妊娠率和活产率均明显高于移植非优质胚胎组,差异有统计学意义(P0.05)。优胚培养液中PAF水平明显高于非优胚培养液,差异有统计学意义(P0.05)。结论:优质胚胎及种植的胚胎可分泌更高水平的PAF,可通过联合胚胎质量和培养液中PAF水平挑选具有种植潜能的胚胎进行移植,从而改善IVF最终结局。     

体外受精实验室空气净化系统高效滤膜更换对体外受精结局影响研究

目的探讨空气净化系统高效滤膜更换对体外受精(IVF)实验室空气中挥发性有机物(VOCs)含量、胚胎质量以及IVF结局的影响。方法回顾性分析2013-12-21—2014-03-13在重庆市妇幼保健院接收IVF助孕且进行移植的周期363个。根据高效滤膜更换时间分为两组:更换前组IVF治疗的235个周期和更换后组128个周期。分析更换前后实验室空气中VOCs含量以及两组正常受精率、可移植胚胎率、优质胚胎率、胚胎种植率、临床妊娠率和流产率等数据。结果滤膜更换前后两组患者的正常受精率分别为69.55%和69.82%;可移植胚胎率分别为55.87%和58.60%;优质胚胎率分别为7.54%和8.06%;胚胎种植率分别为49.14%和49.80%;临床妊娠率分别为62.98%和64.84%;流产率分别为10.14%和10.13%,差异均无统计学意义(P0.05)。结论高效滤膜更换后,实验室空气中VOCs含量明显下降,但对正常受精率、可移植胚胎率、优质胚胎率以及临床妊娠率、种植率和流产率等无明显影响。     

习惯性流产夫妇外周血及绒毛染色体分析

本文分析了１１２例习惯性性流产夫妇外周血染色体及２９例（３８人次）习惯流产妇女再次妊娠流产时绒毛染色体检查结果。结果表明，４例（３．６％）习惯性流产夫妇一方有外周血染色体异常，异常类型均为平衡易位。３８人次绒毛染色体结果中１４例（３６．８％）染色体异常，其中７１．４％为常染色体三体或常染色体三体嵌合体。本结果表明对习惯性流产夫妇进行常规的外周血染色体检查及对流产胚胎进行绒毛染色体检查是十分必要的。     

多囊卵巢综合征患者体外受精/卵胞质内单精子注射-胚胎移植后自然流产相关高危因素的多元回归分析

目的通过了解多囊卵巢综合征(polycystic ovary syndrome,PCOS)患者行体外受精/卵胞质内单精子注射-胚胎移植(IVF/ICSI-ET)助孕后自然流产的发生率及其相关危险因素,探讨PCOS患者IVF-ET后流产的相关危险因素,以为临床改善PCOS患者助孕结局提供理论依据。方法回顾性分析因输卵管因素和/或男方因素行IVF/ICSI-ET助孕的PCOS患者共2231例(PCOS组),对照组为条件匹配的因输卵管因素进行助孕治疗的非PCOS患者共2231例,分析其自然流产率和染色体核型异常率。将PCOS组分为自然流产组和持续妊娠组,利用多元回归分析方法探索PCOS自然流产发生的高危因素。结果 PCOS组的自然流产率(24.15%)较对照组高(12.75%),差异有显著统计学意义(P0.001)。在流产的患者中,PCOS组的染色体核型异常率(36.05%,31/86)较对照组低(55.56%,50/90),差异有统计学意义(P=0.009)。在PCOS患者中,自然流产组的年龄、体质量指数(body mass index,BMI)、HOMA指数水平显著高于持续妊娠组(P0.05);经Logistic回归显示,年龄、HOMA指数及BMI与自然流产的发生密切相关;周期类型、移植胚胎质量、移植胚胎数、空腹血糖、空腹胰岛素及基础性激素等组间均无统计学差异(P0.05)。结论与非PCOS患者相比,PCOS患者的流产率高。染色体核型异常不是PCOS患者高流产率的主要因素。年龄、BMI及HOMA指数是PCOS患者在IVF/ICSI-ET助孕后发生自然流产的危险因素。     

复融新鲜胚胎混合周期移植应用价值探讨

目的:通过分析复融新鲜胚胎混合周期移植的妊娠结局,探寻改善高龄、反复助孕失败患者妊娠结局的方法。方法:回顾分析2014年4月至2016年4月在河南省人民医院生殖医学研究所行常规体外受精/卵胞浆内单精子注射-胚胎移植(IVF/ICSIET)患者的临床资料。根据移植胚胎类型、数量,设置A组(新鲜周期移植D3新鲜胚胎1枚及D3冷冻复苏胚胎1枚,36个周期)、B组(复苏周期移植D3冷冻复苏胚胎1枚,62个周期)和C组(复苏周期移植D3冷冻复苏胚胎2枚,62个周期)、D组(新鲜周期移植D3新鲜胚胎1枚,62个周期)和E组(新鲜周期移植D3新鲜胚胎2枚,62个周期)。比较各组的胚胎种植率、生化妊娠率、临床妊娠率、多胎率和流产率等指标。结果:5组患者中,混合周期患者的既往助孕周期数显著高于其他各组(P<0.05)。5组的妊娠结局、早期流产率、流产率及新生儿出生性别比差异无统计学意义(P>0.05),生化妊娠率、临床妊娠率、胚胎种植率、异位妊娠率、畸形率比较,差异有统计学意义(P<0.05)。A组能获得相对较高的生化妊娠率、临床妊娠率和胚胎种植率。结论:对于可利用胚胎数目较少的高龄反复种植失败的患者,可依据情况建议其采用混合周期移植以改善妊娠结局。     

应用荧光原位杂交技术进行植入前胚胎染色体诊断的价值

目的 初步探讨应用荧光原位杂交(FISH)技术进行植入前胚胎染色体诊断的价值。方法 对10对不孕夫妇进行植入前遗传学诊断(PGD)周期的超促排卵和卵母细胞浆内单精子注射,于受精后第3天进行胚胎活检及FISH分析,第4天选择染色体组成正常或平衡的胚胎进行移植。结果 10个PGD周期共获卵158个,对其中54个胚胎进行活检,51个胚胎获得明确诊断,诊断率为94％(51／54)。对染色体组成正常或平衡的24个胚胎进行官腔内移植,共4例获得妊娠,其中3例已足月分娩健康婴儿,1例为异位妊娠。结论 应用FISH技术进行植人前胚胎染色体诊断,是预防流产和染色体异常患儿出生的有效手段。     

单角子宫IVF-ET助孕双胎妊娠分娩:1例病例报道

目的:报道1例单角子宫患者行IVF-ET助孕双胎妊娠成功分娩活产的病案。方法:对本院接受IVF-ET助孕的1例原发性不孕单角子宫患者进行回顾性总结与分析。结果:采用GnRHa长方案控制性超促排卵(COH),获优质胚胎13枚,移植8cⅡ级胚胎2枚,宫内双胎妊娠因严重子痫前期于34+3周行剖宫产,早产2名健康活婴。结论:对子宫肌层厚度及宫腔容积正常的单角子宫不孕患者行IVF-ET助孕治疗是可行的。但应尽量避免多胎妊娠,减少流产、早产等并发症的发生,争取良好的妊娠结局。     

植入前遗传学诊断四例临床分析

目的 探讨对遗传病高危夫妇采用单细胞荧光原位杂交（FISH）进行胚胎植入前遗传学诊断（PGD）的临床价值。方法 对曾生育过遗传病患儿的4对夫妇通过超排卵获得卵子,体外受精,体外培养至6-8细胞胚胎,每个胚胎取1-2个细胞,采用FISH进行遗传学分析。筛选无遗传病发病风险的胚胎移植入子宫。结果 4例患者共进行4个治疗周期,获得可供活检的胚胎12个,活检细胞20个,固定后有核细胞17个,FISH后除2个细胞无杂交信号外,其余杂交信号清楚,结果明确,活检后的12个胚胎继续发育,结合遗传学诊断,8个胚胎可供移植,其中1例妊娠,于2001年9月14日足月剖宫产分娩一女婴,发育正常,体重4270g,出生后染色体检查为正常女性核型。结论 对遗传病高危夫妇采用FISH技术进行PGD具有临床应用价值。     

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。     

引物延伸预扩增法应用于β-地中海贫血患者胚胎植入前遗传学诊断--附一例报告

目的 探讨对β-地中海贫血进行胚胎植入前遗传学诊断(PGD)的方法。方法 两对夫妇双方均分别为密码子41-42(-TCTT)及插入序列(IVS)-Ⅱ654(c→T)突变杂合子,在本中心进行超排卵和体外受精-胚胎移植治疗,胚胎活检后应用全基因组扩增技术及反向点杂交进行胚胎PGD,根据诊断结果选择健康的胚胎移植入子宫。结果 2例患者共获卵35个,受精率为87％,共活检胚胎16个,获卵裂球25个,单卵裂球总扩增效率为84％,等位基因脱扣率为15％。2例患者共移植5个胚胎,1例获得妊娠,已分娩健康双胎。结论 应用引物延伸预扩增技术可对β-地中海贫血进行PGD,达到优生的目的。     

Preconception and preimplantation diagnosis for cystic fibrosis.

Preimplantation diagnosis provides couples at high genetic risk the possibility of avoiding genetic disease without the need for prenatal diagnosis and selective abortion of the affected pregnancy. Following extensive background work on the reliability of genetic diagnosis in a single cell, we offered on a research basis preimplantation diagnosis to five couples at risk for offspring with the delta-F508 mutation (the major mutation causing cystic fibrosis). There was no detrimental effect from polar body removal on either fertilization or preimplantation development. Genetic analysis, undertaken in 22 polar bodies and 15 corresponding blastomeres, identified 21 embryos of which ten were transferred.     

Pregnancies after in vitro fertilization and transfer of human blastocysts

In a study of 29 cycles of IVF, ET was performed on day 5 after oocyte recovery when embryos had developed to the morula/blastocyst stage. Three preclinical pregnancies and three live births resulted (2 singleton and 1 twin), giving a viable PR per ET of 10%. It is concluded that while day 5 ET may well be important in terms of embryo biopsy for the preimplantation diagnosis of genetic disease, day 2 ET remains preferable for therapeutic IVF. Although these data would not support the introduction of day 5 ET into routine therapeutic IVF, delayed ET should be considered as an alternative approach to preimplantation diagnosis. Indeed, because the latter will generally involve the treatment of normal, fertile couples, it might be predicted that embryo survival rates, and thus the rate of pregnancy after day 5 ET, would be better than those presented here.     

Chromosome analysis in embryos from young patients with previous parity

This study included 173 young couples of proven fertility who had previously undergone preimplantation genetic screening for chromosomes X and Y for family balancing. Several months later, when the outcome of the pregnancies was already known, the blastomeres from the corresponding embryos transferred were reanalysed by fluorescence in-situ hybridization (FISH) for chromosomes 13, 16, 18, 21, 22 with the aim of investigating correlation with embryo viability and the level of FISH sensitivity (embryos confirmed to be euploid). According to the results, informative in 152 couples, the proportion of euploid embryos was significantly lower in 53 nonpregnant women when compared with 99 women with term pregnancy (49% versus 75% respectively, P < 0.001). In addition, in 21 nonpregnant patients, all embryos transferred were found to be chromosomally abnormal. The level of FISH sensitivity was calculated in the group of term pregnancies where the number of euploid embryos was expected to exceed or match with the number of babies born. The resulting false-negative rate was 4.0% per patient and 1.9% per embryo. These findings confirmed the limited prediction power of embryo morphology on implantation but also the relevance of chromosomal abnormalities in causing embryo demise.     

胚胎染色体非整倍体筛查用于平衡易位携带者植入前的遗传学诊断

目的 交信号和1个Y染色体杂交信号者,则诊断为整倍体胚胎;异常杂交信号的胚胎则诊断为非整倍体胚胎.结果 (1)11个平衡易位的PGD周期中,选出杂交信号完整的130个细胞核进行分析,FISH共分析了937个荧光杂交信号,其中整倍体细胞核38个,共有304个杂交信号;其余92个为非整倍体细胞核.(2)在92个非整倍体细胞核中,Ⅰ、Ⅱ及Ⅲ级胚胎的比例分别为20个(22%)、36个(39%)及36个(39%);38个整倍体细胞核中,Ⅰ、Ⅱ及Ⅲ级胚胎的比例分别为13个(34%)、17个(45%)及8个(21%),两者的Ⅰ、Ⅱ及Ⅲ级胚胎数分别比较,差异均无统计学意义(P＞0.05).虽然染色体整倍体率在不同级别胚胎中的分布比较,差异均无统计学意义(P＞0.05),但优质胚胎(Ⅰ级+Ⅱ级)中非整倍体率仍为60%(56/92).(3)平衡胚胎来源的卵裂球细胞核整倍体率(71.4%,30/42)明显高于非平衡胚胎来源的卵裂球细胞核整倍体率(9.1%,8/88),两者比较,差异有统计学意义(P＜0.05).平衡胚胎来源的卵裂球细胞核非整倍体率(包括三体、单体、复杂非整倍体、单倍体、多倍体)明显低于非平衡胚胎来源的卵裂球细胞核非整倍体率(P＜0.05).结论 平衡易位携带者的胚胎中有较高的非整倍体率,因此,胚胎非整倍体筛查在平衡易位携带者的PGD中有重要价值和临床意义.

Abstract：

Objective To determine the importance of aneuploidy screening in preimplantation genetic diagnosis for the couples of chromosome translocation carriers. Methods To perform 11 prenatal genetic disgnosis (PGD) cycles for 7 couples of chromosome translocation carriers from January 2006 to March 2009 in the Reproductive Medical Center, First Affiliated Hospital of Zhengzhou University. To re-analyze the well-fixed, non-multinuclear and non-debris nuclei using the probes of LSI 13, 18, 21,CEPX, CEPY to detect the aneuploidy rate which come from the PGD cycles of the couples of chromosome translocation carriers. The euploid embryo was defined as two fluorescence in situ hybridization (FISH)signals of LSI 13, 18, 21 respectively and two signals of CEPX, or one signal of CEPX and one signal of CEPY. The other abnormal signals were defined as aneuploid embryo. Results (1) A tolal of 130 nuclei from 11 PGD cycles got the integrated re-FISH signals. Nine hundred and thirty-seven FISH signals were analysized, including 304 signals from 38 euploid nuclei and the others from 92 aneuploid nuclei. (2) The number of the aneuploid nuclei from grade Ⅰ , Ⅱ and Ⅲ embryo was 20 (22%), 36(39%), and 36(39%). The number of the euploid nuclei from grade Ⅰ , Ⅱ and Ⅲ embryo was 13(34%), 17(45%),and 8(21%). There was no significant difference of aneupioidy rate between the embryos form different grades (P＞0.05). However, the rate of aneuploid nucleus from good quality embryos (grade Ⅰ + grade Ⅱ) was 60% (59/92). (3) The euploidy rate was 71.4% (30/42) from balanced embryos, while 9.1%(8/88)from unbalanced embryos. There was significant difference between them (x2=53.4, P＜0.05).The rate of aneuploidy from balanced embryos was lower than those from unbalanced embryos (P＜0.05).Conclusions Since higher rate of aneuploidy was detected in embryos of the couples of chromosome translocation carriers. It is advisable to recommend the FISH re-analysis for aneuploidy screening to preimplantation genetic diagnosis for the couples of chromosome translocation carriers.     

The role for preimplantation genetic diagnosis in balanced translocation carriers

OBJECTIVE: Preimplantation genetic diagnosis is an established technique that provides an alternative to prenatal diagnosis for patients who are at risk of transmitting a serious genetic disorder to their offspring. Preimplantation genetic diagnosis has been used for couples who have been at risk for having offspring with single gene or X-linked disorders and for screening for common age-related aneuploidy and in couples who themselves carry balanced chromosomal rearrangements. The aim of this study was to summarize our experience using preimplantation genetic diagnosis after the identification of a parental balanced translocation, specifically as it relates to the number of embryos that are suitable for transfer after preimplantation genetic diagnosis for a known translocation and aneuploidy screening. STUDY DESIGN: This is a retrospective review of data from a single center that involved 6 couples that initiated the process of preimplantation genetic diagnosis for translocation and aneuploidy screening by fluorescent in situ hybridization. RESULTS: A total of 65 embryos were obtained, of which 56 embryos (86%) were suitable for fluorescent in situ hybridization analysis. After fluorescent in situ hybridization, 1 embryo was diagnosed as normal or balanced (1.7%). Forty-three embryos (76.8%) were unbalanced for the translocation; 8 embryos (14.3%) were aneuploid, and 4 embryos (7.1%) were uninformative. There were no clinical pregnancies. CONCLUSION: In our experience, there are very few embryos that are available for transfer from these patients after translocation and aneuploidy screening because of multiple unbalanced segregation products and a high rate of aneuploidy. Factors that contributed to this may be related to which parent carries the translocation, methods that were used for in vitro fertilization, and advanced maternal age. Although preimplantation genetic diagnosis for translocation carriers theoretically can enhance the pregnancy rate for a couple, there are limitations. This information should be shared with couples who are contemplating preimplantation genetic diagnosis for translocation, and the options of sperm or egg donor should be considered.     

Pregnancy after polar body biopsy and freezing and thawing of human embryos

OBJECTIVE: To evaluate the outcome of frozen-thawed ET using embryos previously biopsied for preimplantation genetic diagnosis during a fresh ET cycle. DESIGN: Prospective evaluation. SETTING: Assisted reproductive biology program. PATIENT(s): A 31-year-old, G4, P1, TAB1, SAB2 carrier of a balanced RT 45,XX der(14;21)(q10;q10) translocation. INTERVENTION(s): Preimplantation genetic diagnosis by polar body biopsy. Excess embryos were frozen using the one-step method and then thawed. MAIN OUTCOME MEASURE(s): Embryo survival after thawing and subsequent pregnancy outcome. RESULT(s): Among the 32 mature oocytes, the results of fluorescence in situ hybridization were available for 25 polar bodies. Eleven were unbalanced, 10 were normal (8 fertilized), and 4 were balanced (3 fertilized) for the fresh IVF cycle. Two normal embryos were transferred. Four normal and 3 balanced embryos were cryopreserved. A chemical pregnancy resulted. Four months later, the 7 cryopreserved embryos were thawed; 2 survived (1 balanced and 1 normal) and were transferred. An ongoing pregnancy resulted, and a normal (46,XX) female was delivered. CONCLUSION(s): Freezing and thawing of biopsied embryos resulted in a low survival rate. However, this should not be a deterrent to the cryopreservation of extra chromosomally normal embryos because the embryos that do survive are able to implant.     

First successful pregnancy following PGD for chromosome translocation on embryos generated from in-vitro matured oocytes: a case report

As far as is known, this is the first report of a successful pregnancy outcome following preimplantation genetic diagnosis for a chromosome translocation in embryos generated from in-vitro matured oocytes. A couple presented to the study clinic where the female partner was a carrier of the reciprocal chromosome translocation 46,XX,t(1;20)(p36.1;p12.2) with three consecutive pregnancy terminations due to either fetal abnormality or unbalanced translocation products detected in the conceptus. Under routine ultrasound investigation she was diagnosed with polycystic ovaries. The patient underwent an in-vitro maturation/preimplantation genetic diagnosis cycle where the immature oocytes were matured in vitro and fertilized by intracytoplasmic sperm injection. Day-3 embryos were screened for the chromosome abnormality by fluorescent in-situ hybridization. A single embryo diagnosed as chromosomally normal/balanced was transferred on day 5 and resulted in the birth of a healthy child.