2178对自然流产史夫妇染色体分析

目的:分析自然流产史夫妇外周血染色体异常核型的种类及其在男女性中的分布特点。方法:检测2178对自然流产史夫妇外周血淋巴细胞染色体核型,分析比较染色体变异的种类、发生率及其在男、女性中的分布差异。结果:2178对(4356例)自然流产史夫妇中发现染色体异常539例(12.37%),其中男266例,女273例。染色体结构异常87例,其中相互易位最多见为58例(66.7%,58/87),有27例为世界首报染色体结构异常核型。其次为罗伯逊易位13例,倒位6例,插入、缺失等其他异常核型10例。染色体数目异常8例,包括2例标记染色体、1例XYY及5例不同类型的X染色体非整倍体嵌合。多态性改变444例中,D/G组染色体随体区变异最为多见,共271例(61.04%,271/444)。结论:自然流产史夫妇外周血染色体异常均有发生,男女性发生率并无明显差异,染色体异常以相互易位为主。染色体多态性发生率较高,对以自然流产史就诊的夫妇,有必要同时进行染色体检查,有助于病因的分析与诊断,并为临床咨询及后续生殖干预提供依据。     

Cytogenetic study in 50 couples with recurrent abortions

The presence of chromosome abnormalities in couples with repeated spontaneous abortion is known even if the phenomenon is far from a complete assessment. A cytogenetic investigation in 50 couples with a history of two or more spontaneous abortions is referred to in this study. A peripheral blood lymphocyte culture was harvested for each subject and the slides were stained by G- and C-banding. Of the 100 individuals examined, 4 were carriers of balanced translocations, 3 of which were of the Robertsonian type. A chromosomal fragility (chromatidic and/or chromosomic gaps) was seen in 2 cases. The incidence of balanced translocations found here is 8% which is near to the mode (about 9%) observed in previous studies. Those frequencies are greater than in the general population (0.1-0.4%). This indicates that balanced translocations have some importance in causing abortion while this is not the case for other chromosomal abnormalities (e.g. pericentric inversions). Thus, cytogenetic analyses should be recommended in couples with repeated spontaneous abortions, when clinical data fail to clarify the cause.     

Cytogenetic findings in fifty-five couples with recurrent fetal wastage

Balanced chromosomal translocations in parents and Müllerian abnormalities constitute defined causes of reproductive wastage. Fifty-nine couples with histories of recurrent abortion with or without fetal malformations were evaluated with cytogenetic studies and gynecography. In 44 of the couples with pure abortion histories of two or more spontaneous abortions, three (6.8%) balanced carrier parents were identified. In 11 couples with a mixed history of abortion plus fetal malformation, 3 (27.3%) had balanced translocations in one of the parents. The over-all incidence of Müllerian abnormalities in the group of 59 patients was 11.9%.     

两种不同遗传学分析方法用于诊断自然流产组织的比较

目的探讨比较基因组杂交（CGH）技术与绒毛细胞培养染色体核型分析用于自然流产组织遗传学诊断的准确性。方法选择妊娠49—91d的自然流产患者38例,在无菌条件下经宫颈取绒毛,其中难免流产的新鲜组织标本27份,过期流产的陈旧组织标本11份。每份组织标本均采用绒毛细胞培养染色体核型分析,并同时采用CGH技术对全基因组进行分析。结果CGH技术诊断成功率为100％（38／38）,而绒毛细胞培养染色体核型分析诊断成功率为82％（31／38）。两种方法的诊断符合率为90％（28／31）,在3例出现不同诊断结果的病例中,1例绒毛细胞培养染色体核型分析显示染色体核型正常,而CGH技术显示3q^22_q^24缺失;另2例绒毛细胞培养染色体核型分析为3倍体,但CGH技术诊断结果显示正常。在7例绒毛细胞培养失败而仅有CGH技术诊断结果者中,3例为染色体非整倍体异常,另4例正常。结论CGH技术用于诊断自然流产组织是可行的。绒毛细胞培养染色体核型分析比较,CGH技术诊断成功率高,且对非平衡染色体结构重排的诊断有较高的敏感性,可以作为绒毛细胞培养染色体核型分析的补充方法。     

不同年龄和流产次数的复发性流产患者绒毛染色体核型分析

目的:了解复发性自然流产胚胎染色体异常发生情况。方法:2008年1月至2011年12月,在我院诊治并成功行绒毛染色体核型分析的自然流产患者235例,根据自然流产次数分为复发性流产组(125例)和偶发性流产组(110例)。比较两组绒毛染色体异常发生率和类型的差异,不同流产次数的胚胎染色体异常发生情况,以及不同年龄患者绒毛染色体异常的发生情况。结果:复发性流产组,绒毛染色体异常发生率显著低于偶发性自然流产组(47.2%vs 70.9%,P<0.05)。复发性流产组中三体占异常染色体的66.1%(39/59),显著高于偶发性流产组(44.8%,35/78)(P<0.05)。随着自然流产次数的增加,绒毛染色体异常发生率降低,差异有统计学意义(χ2=15.266,P=0.004)。复发性流产组中,年龄≥35岁者的绒毛染色体异常发生率明显高于年龄<35岁者(60.9%vs39.2%,P<0.05)。偶发性自然流产组中,年龄≥35岁者的绒毛染色体异常发生率亦明显高于年龄<35岁者(88.9%vs 62.2%,P<0.05)。结论:胚胎染色体异常是引起复发性流产的一个重要原因,随着流产次数的增加,流产胚胎染色体异常的发生率降低。无论是复发性流产还是偶发性流产,高龄均是引起胚胎染色体异常的高危因素。     

Cytogenetic studies of spontaneous abortions in humans

Karyotypes were analyzed using directing method for chorionic villi chromosome preparation in 52 specimens from spontaneous abortions. A paralleled cytogenetic study of 51 specimens from induced abortions was carried out. Among 52 specimens from spontaneous abortions, 11 (21%) were chromosomally abnormal and 82% of these were found to have autosomal trisomies (3,14,16,19,20 and 21). Trisomy 16 was the most common abnormality accounting for 44% of trisomic abortuses. A double trisomic abortus with a 48, XX, +16, +20 karyotype was detected. These results showed that chromosome abnormality is an important cause of spontaneous abortion. The relationship between chromosomal abnormalities of spontaneous abortuses and the factors relevant to the abnormalities are discussed.     

染色体异常与自然流产

染色体异常是自然流产的常见原因,包括夫妇染色体异常和胚胎染色体异常。其中夫妇染色体异常涉及结构异常,如相互易位、罗氏易位、倒位等;数目异常,如特纳综合征、克氏综合征等。胚胎染色体异常以非整倍体为主。文章将对染色体异常与自然流产的相关研究进展予以阐述。     

染色体异常与自然流产

染色体异常是自然流产的常见原因,包括夫妇染色体异常和胚胎染色体异常。其中夫妇染色体异常涉及结构异常,如相互易位、罗氏易位、倒位等;数目异常,如特纳综合征、克氏综合征等。胚胎染色体异常以非整倍体为主。文章将对染色体异常与自然流产的相关研究进展予以阐述。     

Pregnancy outcomes in unselected singleton pregnant women with an increased risk of first-trimester Down's syndrome

BACKGROUND: The purpose of this study was to assess outcomes in pregnancies with a positive screen of first-trimester combined test (nuchal translucency, pregnancy-associated plasma protein-A and free beta-human chorionic gonadotropin). METHODS: Using a cut-off level of 1 in 270, 216 (7.1%) women had a positive screen. Among them, 187 delivered their babies in our hospital and the adverse outcomes, such as spontaneous abortion, intrauterine fetal demize, preterm prelabor rupture of the membranes, preterm labor, intrauterine growth restriction, gestational hypertensive disorders, placenta previa, chromosomal abnormalities and fetal structural anomalies, were identified and compared with the 2097 women who screened negative for Down's syndrome. RESULTS: Pregnancies with a positive screen had a significantly higher risk of adverse outcomes than those with negative screens (30.5% versus 15.3%; odds ratio 2.4; p < 0.001), especially for those complicated by spontaneous abortion (odds ratio 11.4; p < 0.05) and placenta previa (odds ratio 4.3; p < 0.05). CONCLUSIONS: Besides fetal chromosomal abnormalities and structural abnormalities, pregnancies with a positive screen for Down's syndrome in the first-trimester had a significantly higher incidence of subsequent adverse obstetric outcomes.     

Array comparative genomic hybridization profiling of first-trimester spontaneous abortions that fail to grow in vitro

OBJECTIVES: Cytogenetic analysis of spontaneous abortion samples can be limited by culture failure. Failure to grow in vitro has traditionally been suspected to be due to in vivo death of tissue associated with spontaneous abortion (SAB) or simply technical factors of growth in culture. METHOD: We used array comparative genomic hybridization (array CGH) to investigate chromosomal imbalances in products of conception that failed to grow in vitro. RESULTS: Our data on 26 cases of SABs that failed to grow in culture are compared and contrasted with published data on cytogenetic findings following in vitro culture. The results revealed abnormalities uncommonly seen by classic cytogenetic methods. These abnormalities include high rates of double aneuploidy and autosomal monosomy. The data taken together suggest that classic cytogenetics of spontaneous abortion may yield normal karyotypes or selected abnormal karyotypes that permit cell proliferation in vitro while Array CGH detects other abnormalities. CONCLUSION: Array CGH is becoming an important clinical assay for unbalanced chromosome abnormalities whether cells grow in culture or not and in cases of analysis on one or few cells.     

Survey of 1120 Japanese women with a history of recurrent spontaneous abortions.

Our recent 7-year clinical survey showed that among the 1120 women with repeated spontaneous abortions registered in this clinic, 2898 out of a total of 3216 pregnancies (90.1%) had terminated in spontaneous abortion. Among these wastages, 84.2% occurred before 12 weeks of gestation, and 11.1 percent occurred between 12 and 15 weeks. Through routine examination of reproductive wastage, 82 (9.9%) of the 825 Japanese couples examined were shown to have either a chromosomal abnormality or normal variants in the wife and/or husband, thus demonstrating no racial difference in the incidence of chromosomal abnormalities in infertile patients in comparison with studies performed in other countries. One hundred and forty-seven congenital uterine anomalies (14.7%) were found in 1000 hysterosalpingographies, and 12 of 148 examined females were positive for anti-cardiolipin antibody. 393 other females with no major abnormalities likely to induce spontaneous abortions were indicated for immunotherapy. Ample time spent on genetic counseling prevented further reproductive wastage, and ideal metroplasty resulted in a successful post-operative pregnancy course in more than 85% of cases. Immunosuppressant and anticoagulant therapy decreased the serum titer of anti-cardiolipin antibody, enabling pregnancies to be maintained to term. Immunotherapy utilizing the husband's lymphocytes also brought more than 80% of pregnancies to successful completion, with 200 deliveries achieved with this therapy. In contrast, 64.1% of pregnancies again terminated spontaneously in patients who were indicated for immunotherapy but did not receive treatment. The findings of the present study suggest that the causes of reproductive wastage, especially the etiology of early recurrent spontaneous abortion, are complex.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pregnancy outcomes with increased nuchal translucency after routine Down syndrome screening.

OBJECTIVES: The purpose of this study was to evaluate the outcomes of pregnancies with nuchal translucency greater or equal to 3 mm for routine first trimester screening in unselected populations. METHODS: A total of 2980 pregnant women for first trimester ultrasonography were routinely offered crown-rump length (CRL) and nuchal translucency (NT) for screening for Down syndrome between 11 and 14 weeks' gestation. A complete follow-up was obtained in all cases by a review of medical records. RESULTS: Using a cut-off value of 3 mm, the prevalence of increased fetal NT was 0.7% (n=22). Among the 22 cases, there were five (22.7%) chromosomal abnormalities. Of the 17 chromosomally normal pregnancies, four resulted in fetal demise (spontaneous abortion, intrauterine death or termination of pregnancy due to fetal abnormalities). The remaining 13 pregnancies resulted in live births, including one gestational hypertension and one preterm delivery, respectively. The total incidence of an adverse outcome in the group of increased fetal NT was 45.5%. CONCLUSIONS: In a routine population with first-trimester ultrasonography, fetal NT measuring greater than or equal to 3 mm was associated with a poor pregnancy outcome with not only chromosomal abnormalities and congenital cardiac diseases, but also poor maternal and fetal health or adverse pregnancy outcomes. In addition, this study also demonstrated the necessity for fetal assessment and follow-up in cases where the fetal NT is increased in the first trimester.     

拷贝数变异测序在染色体易位夫妇植入前遗传学诊断中的临床应用

目的探讨拷贝数变异测序(copy number variation sequencing, CNV-seq)用于染色体易位夫妇胚胎植入前诊断的应用价值。 方法回顾性分析2017年1月至2018年12月,在广东省妇幼保健院生殖健康与不孕症科进行植入前遗传学诊断(preimplantation genetic diagnosis,PGD)的211对染色体易位夫妇患者的临床病例。使用CNV-seq对胚胎染色体进行检测,并对患者一般信息和PGD结果进行分析。 结果1210个胚胎中,被检出837个(79.2%)胚胎存在染色体异常,373个(30.8%)胚胎为整倍体。在241个PGD周期中,68个(27.6%)周期所有胚胎均存在染色体异常,178个(72.4%)周期至少含有一个整倍体胚胎。在176个移植周期中,130个(73.9%)确定临床妊娠,已出生46个健康婴儿,12例发生早期流产。 结论CNV-seq可准确地区分胚胎染色体是否存在异常,避免因胚胎含有染色体异常而被移植,是一种可靠而准确的PGD技术。     

Effect of maternal age on spontaneous abortion during the first trimester in Northeast China

Objective: We aimed to determine the relationship between fetal chromosomal abnormalities and maternal age among spontaneous first trimester abortions in women in Northeast China.

Methods: We evaluated 497 chorionic villi samples from patients with a history of spontaneous abortion during the first trimester. We divided the samples into five groups according to the maternal age: <25, 25–29, 30–34, 35–39, and ≥40 years. We identified chromosomal abnormalities by fluorescence in situ hybridization.

Results: Among the 497 chorionic samples of spontaneous abortion, 180 (36.22%) had fetal chromosomal abnormalities. Patients aged ≥40 years had a significantly higher percentage (60.61%) of fetal chromosomal abnormalities compared with the other groups. More women in the ≥40 and 35- to 39-year groups had a history of three consecutive miscarriages and 10 kinds of abnormalities. The most frequent aneuploidy was trisomy 22, followed by trisomy 16.

Conclusions: These results revealed that the kinds of fetal abnormalities, numbers of abortions, and chromosomal abnormality rates increased with increasing maternal age. The most common trisomy types in spontaneous abortions were closely related to maternal age. We hypothesize whether the larger probability of chromosomal abnormalities is due to increased mutation rate with maternal age, or due to a worse in-utero conditions.     

Quantitative fluorescent polymerase chain reaction to detect chromosomal anomalies in spontaneous abortion

OBJECTIVES: To evaluate the value of short tandem repeats (microsatellites) in the study of numerical chromosomal anomalies in spontaneous abortion. METHOD: Multiplex quantitative fluorescent polymerase chain reaction (QF-PCR) was carried out on 61 spontaneous abortion samples and 48 controls using microsatellite markers from 8 chromosomes where aneuploids are commonly found. RESULTS: Of the 61 samples, 65.6% were successfully karyotyped, and the call rate of the QF-PCR was 98.3%. The correspondence between PCR and karyotyping was 95%. The success rate of karyotyping in the inevitable abortion group was 79.6%, higher than for the missed abortion group (8.3%), P<0.001. The call rate of QF-PCR showed no difference between these 2 groups (100% vs 91.7%, P=0.197). CONCLUSION: Microsatellite-based QF-PCR is a helpful and reliable tool to diagnose numerical chromosomal anomalies in spontaneous abortion. It also provides a diagnosis for necrotic tissue.     

Cytogenetic findings and reproductive outcome of infertile couples referred to an assisted reproduction program

It is still undefined whether all the couples entering an assisted reproduction program should undergo to karyotype analysis. The present study was conducted to determine the prevalence of chromosomal abnormalities in a non-selected sample of 1,146 couples referred to assisted reproduction technologies (ART), and to analyze the outcome of pregnancies from couples in whom cytogenetic anomalies were detected. Irrespective of the infertility factor, fertilization was achieved by intracytoplasmic sperm injection (ICSI). A total number of 35 karyotype anomalies were diagnosed, corresponding to an abnormality frequency of 1.52% (1.83% for men and 1.22% for women). As could be expected, the majority of men presenting katyotype anomalies had a low sperm count. Among women, the majority of cytogenetic anomalies were detected in individual not presenting risk factors for aberrant karyotype. Around 41% of pregnancies achieved in couples presenting chromosomal anomalies ended in spontaneous abortion. Information on fetal karyotype was limited. No major malformations were observed among newborns from parents with abnormal karyotype. In consideration of the elevated frequency of pregnancy loss, it seems advisable to recommend that chromosomal analysis be performed in all couples undergoing ART. This with the aim of identifying patients that would possibly benefit from pre-implantation genetic diagnosis.     

Chromosomal copy number analysis of products of conception by conventional karyotyping and next-generation sequencing

PurposeChromosomal abnormalities are a major cause of spontaneous abortion, and conventional G‐banded karyotyping (G‐banding) is mainly utilized for chromosomal analysis. Recently, next‐generation sequencing (NGS) has been introduced for chromosomal analysis. Here, we aimed to investigate the applicability and utility of NGS‐based chromosomal analysis of products of conception (POC) on chorionic villus samples from spontaneous abortion.MethodsThe results of chromosomal analysis of 7 chorionic villus samples from spontaneous abortion were compared between conventional G‐banding and NGS‐based chromosomal copy number analysis. Age dependency and frequency of each chromosomal aneuploidy were evaluated for 279 cases analyzed by NGS.ResultsExcluding two cases (culture failure and maternal cell contamination), the results were consistent between G‐banding and NGS. For cases analyzed by NGS, the rate of chromosomal abnormality increased in a maternal age‐dependent manner. The frequency of each chromosomal aneuploidy detected by NGS was almost the same as that previously reported. Finally, NGS analysis was possible for difficult cases by G‐banding analysis, such as culture failure, maternal cell contamination, long‐term storage cases, and low cell number.ConclusionsChromosome analysis using NGS not only obtains comparable results to conventional G‐banding, but also can analyze POC more accurately and efficiently.     

自然流产夫妇的细胞遗传学研究─附10例世界首报染色体核型

用人体外周血淋巴细胞培养法对６２４对原因不明的自然流产夫妇进行了细胞遗传学研究，检出异常染色体携带者５７例，其中女３８例，男１９例。５７例染色体异常中，结构异常４５例；染色体数目异常２例；９号染色体臂间倒位９例，Ｙ染色体倒位１例。异常染色体涉及到１，３，４，５，６，７，８，９，１０，１１，１３，１４，１５，１７，１８，１９，２１，２２，Ｘ，Ｙ共２０条。４５例结构异常中１０例经湖南医科大学国家细胞遗传学培训中心鉴定为世界首报。     

自然流产夫妇的细胞遗传学研究─附10例世界首报染色体核型

用人体外周血淋巴细胞培养法对６２４对原因不明的自然流产夫妇进行了细胞遗传学研究，检出异常染色体携带者５７例，其中女３８例，男１９例。５７例染色体异常中，结构异常４５例；染色体数目异常２例；９号染色体臂间倒位９例，Ｙ染色体倒位１例。异常染色体涉及到１，３，４，５，６，７，８，９，１０，１１，１３，１４，１５，１７，１８，１９，２１，２２，Ｘ，Ｙ共２０条。４５例结构异常中１０例经湖南医科大学国家细胞遗传学培训中心鉴定为世界首报。     

Fertility in women after age forty-five.

Pregnancy after age 45 years is infrequent and the mother and baby should be considered as a high risk. There is a greater incidence of spontaneous abortion, gestational trophoblastic disease and chromosomal abnormalities in the fetus. Birth control practices should be discontinued after 49 years of age in the best interest of the woman's sexuality if abortion is acceptable to her. The patient should be completely informed of the risks and, in the event of pregnancy, abortion should be advised.