无创产前基因检测在胎儿染色体非整倍体筛查中的应用研究

目的:探讨无创产前基因检测(NIPT)在胎儿染色体非整倍体疾病诊断中的临床应用价值及意义。方法:收集2016年1月至2016年12月在我院行NIPT检测的6030例单活胎孕妇(孕12~32周)外周血标本并提取胎儿游离DNA,进行文库构建后利用高通量测序技术,结合生物学信息分析,推算出胎儿患染色体疾病的风险率。对检测结果提示高风险者建议行羊水或者脐血穿刺及染色体核型分析;对检测结果低风险者进行电话跟踪随访。结果:行NIPT检测的6030例孕妇检出高风险67例,其中21-三体高风险16例,18-三体高风险3例,13-三体高风险5例,性染色体异常高风险34例,其他染色体异常高风险9例。67例高风险孕妇中,10例拒绝进一步产前诊断,余57例均行羊水/脐血穿刺检查,其中确诊胎儿染色体异常34例(21-三体16例,18-三体2例,13-三体2例,性染色体异常12例,其他染色体异常2例);余23例羊水/脐血穿刺结果正常者,继续妊娠。NIPT对21-三体和18-三体的阳性预测值达100.00%和66.67%,但对13-三体、性染色体异常以及其他染色体异常的阳性预测值仅50.00%、42.85%和33.33%。结论:NIPT对21-三体和18-三体的检测符合率较高,但对13-三体、性染色体异常以及其他染色体异常符合率偏低。NIPT是一项筛查技术,测序高风险的孕妇仍需行羊水/脐血穿刺,以保证产前诊断的准确性。     

无创产前检测进行染色体缺失或重复检测的临床应用价值

目的:探讨无创产前检测(NIPT)进行染色体缺失或重复检测的临床应用价值。方法:对传统产前筛查异常、既往有唐氏儿分娩史及高龄孕妇等3500例孕妇采集外周血,采用Illumina测序技术检测母体血浆胎儿游离DNA(cff DNA),分析胎儿性染色体及除21、13、18号染色体以外的其他常染色体信息,对NIPT阳性的孕妇进行遗传咨询,对其中自愿进行介入性产前诊断的,行染色体核型分析及染色体微阵列分析(CMA)进行验证。结果:3500例接受NIPT的病例中,检出32例常染色体异常(除外21、13、18号),其中有11例接受介入性产前诊断,染色体核型分析及CMA检测分别提示3例异常,符合率27.3%(3/11)。检出45例性染色体异常,其中有23例接受介入性产前诊断,染色体核型分析及CMA检测分别确诊12例异常,符合率52.2%(12/45)。结论:NIPT在预测胎儿性染色体异常及常染色体(除外21、13、18号)异常方面有一定的参考价值,但需要进行染色体核型分析和(或)CMA检测进行验证。     

8594例孕妇无创DNA产前检测在胎儿染色体非整倍体疾病筛查中的临床应用

目的:分析无创DNA产前检测(NIPT)技术在胎儿染色体非整倍体疾病筛查中的临床应用价值。方法:选择2014年10月至2017年12月在空军军医大学第一附属医院妇产科门诊接受NIPT的8594例孕妇为研究对象,统计不同年龄段、孕周的孕妇染色体异常风险增加情况,并进行NIPT与羊水细胞染色体核型分析的结果符合情况比较。结果:8594例孕妇筛查出染色体异常高风险96例(1. 12%)。筛查出21-三体综合征、18-三体综合征、13-三体综合征高风险44例(0. 51%),性染色体异常高风险45例(0. 52%)和其他常染色体异常高风险7例(0. 08%)。年龄≥35岁孕妇染色体异常高风险(1. 80%),高于30岁(0. 72%)和30～34岁(1. 19%)年龄段,差异有统计学意义(P 0. 05)。8594例孕妇中于孕中期(孕13～20周)接受NIPT的孕妇比例最高(6556例,76. 29%),经羊水细胞染色体核型分析确诊率也最高(42/61,68. 85%)。77例NIPT染色体异常高风险孕妇经羊水细胞染色体核型分析确诊54例(70. 13%),其中阳性预测值分别为21-三体综合征100. 00%、18-三体综合征62. 50%、13-三体综合征25. 00%、性染色体异常54. 55%、其他常染色体异常50. 00%。结论:NIPT对21-三体综合征具有较高的检测价值。选择孕中期进行NIPT准确性较高。NIPT可以作为高龄孕妇的主要筛查手段,能有效地避免染色体异常患儿的出生。     

无创DNA检测在诊断高龄孕妇胎儿非整倍体中的应用

目的:探讨高龄妊娠胎儿染色体异常的风险以及无创DNA产前检测(NIPT)在诊断高龄孕妇胎儿非整倍体染色体病中的应用价值。方法:选择≥35岁的高龄孕妇2714例,按年龄分为35~39岁,≥40岁两组,采用NIPT高通量测序检测孕妇血浆游离DNA,并对检测结果提示21-三体、18-三体、13-三体及性染色体高风险者行羊膜腔穿刺术及胎儿染色体核型分析,对检测结果阴性者通过电话随访进行验证。计算NIPT检测的敏感度、特异度、阳性预测值、阴性预测值及Youden指数。结果:2714例高龄孕妇NIPT检测结果提示胎儿非整倍体染色体异常高风险47例,6例高风险孕妇拒绝侵入性产前诊断,余41例高风险孕妇行羊膜腔穿刺术及羊水细胞染色体核型分析,结果显示21-三体19例,18-三体1例,13-三体2例,性染色体异常7例。与现有的金标准羊膜腔穿刺术核型分析相比较,NIPT对高龄孕妇除外性染色体异常的胎儿非整倍体染色体异常检出敏感度为100.00%,特异度为99.93%,阳性预测值为90.91%,阴性预测值为100.00%,Youden指数为0.99。进一步通过年龄分组发现,40岁及以上年龄组异常率显著高于35~39岁年龄组(P=0.011)。结论:高龄孕妇可通过NIPT快速、安全地筛查出胎儿非整倍体染色体异常,减少侵入性产前检测比例,降低胎儿出生缺陷率。     

无创产前基因检测在胎儿性染色体疾病筛查中的应用

目的探讨无创产前基因检测(non-invasive prenatal genetic testing,NIPT)在胎儿性染色体疾病中的临床价值。方法统计深圳市第二人民医院2016年3月至2019年6月行NIPT的16119例单胎孕妇中提示21-三体、18-三体、13-三体、性染色体异常和其他染色体异常的阳性率;对比2016年3月至2019年6月NIPT示“胎儿性染色体异常”与本院染色体核型结果。结果①NIPT对21-三体,18-三体,13-三体,性染色体异常和其他染色体异常筛查的阳性率分别为0.42%(68/16119),0.10%(16/16119),0.07%(11/16119),0.38%(61/16119)和0.22%(36/16119)。②47例“NIPT示胎儿性染色体异常”者中,确诊为性染色体疾病者26例,阳性预测值55.32%。其中,NIPT对胎儿性染色体数目偏多的阳性预测值为91.30%(21/23),对胎儿性染色体数目偏少的阳性预测值20.0%(4/20),对胎儿性染色体数目异常的阳性预测值25.0%(1/4)。结论NIPT可作为胎儿性染色体疾病的筛查方法,但由于其对性染色体偏少和性染色体数目异常的假阳性率较高,检测阳性者仍需要做侵入性产前诊断确诊。     

孕妇外周血胎儿游离DNA无创产前检测应用价值探讨

目的:研究胎儿游离DNA无创产前检测(NIPT)胎儿非整倍体筛查的准确性。方法:选取2013年8月至2017年2月于我院行羊水穿刺及胎儿染色体检查的孕妇共800例,孕妇均已获得外院NIPT检测结果,其中NIPT阳性557例,包括21三体高风险256例,18三体高风险85例,13三体高风险37例,性染色体异常85例,其他染色体异常94例;NIPT阴性243例,包括超声畸形筛查发现胎儿结构畸形或软指标183例,高龄27例,上胎异常7例,早孕用药1例,无指征要求做羊水穿刺25例。NIPT检测结果来自国内11家医院或检测机构。染色体检查采用核型分析和染色体芯片分析(CMA),分析比较800例孕妇的有创产前诊断与NIPT无创产前诊断结果。结果:NIPT对21三体、18三体、13三体和性染色体非整倍体的阳性预测值分别为81.6%(209/256),68.2%(58/85),8.1%(3/37)和41.2%(35/85),NIPT对21三体的阴性预测值为99.6%(242/243),对13三体、18三体和性染色体非整倍体的阴性预测值均为100%(243/243);NIPT对其他染色体非整倍体的阳性预测值为1.6%(1/61),NIPT对CNV的阳性预测值为54.5%(18/33)。结论:分散在各家检测机构模式下的NIPT检测结果阳性预测值较低,NIPT的临床应用需加强规范和质控。     

高通量测序胎儿染色体非整倍体筛查方法在双胎妊娠中的应用

目的探讨高通量测序用于胎儿染色体异常产前筛查在双胎妊娠胎儿非整倍体染色体异常产前检测中的应用价值。方法选择妊娠15~18周接受孕中期血清学筛查和核型分析的200例双胎孕妇,妊娠12~27周行非侵入产前诊断(NIPT),对于染色体异常风险度升高者行双胎妊娠羊膜腔穿刺染色体核型分析;未提示风险度升高者,分娩时留取脐血检查有无染色体异常;双胎妊娠出现流产等不良妊娠结局者,留取胎儿组织提取DNA等进行高通量测序及染色体核型分析。以胎儿/新生儿染色体核型分析结果为金标准,分析NIPT结果。结果200例双胎妊娠患者,行孕中期血清学筛查155例(77.5%,155/200),45例(22.5%,50/200)因年龄高风险未行血清学筛查。155例血清学筛查中,高风险孕妇13例(8.4%,13/155),核型分析和妊娠结局均未发现异常。200例NIPT显示,1例性染色体异常(0.5%,1/200),染色体核型分析证实双胎之一胎儿核型为45,XO;1例18-三体综合征高风险(0.5%,1/200),双胎核型未见明显异常;1例21-三体高风险(0.5%,1/200),双胎之一核型为47,+21,XN;余197例染色体核型分析未提示明显异常。结论高通量测序胎儿染色体异常产前筛查在双胎胎儿染色体非整倍体的检测有一定的应用价值。     

染色体微阵列分析在无创产前筛查非整倍体高风险中的应用价值

目的:探讨染色体微阵列分析(CMA)技术在无创产前筛查(NIPT)提示非整倍体高风险胎儿的产前诊断中的应用价值。方法:241例NIPT提示胎儿染色体非整倍体高风险的孕妇,经羊膜腔或绒毛穿刺获取胎儿细胞,进行CMA分析。对于NIPT提示21-三体和性染色体非整倍体高风险的孕妇根据筛查原因(高龄筛查组、唐筛异常或超声软指标异常组、非高龄筛查组)进行组间NIPT阳性预测值的比较,并对CMA检测提示有染色体拷贝数变异(CNV)的病例进行致病性分析。结果:NIPT对于染色体非整倍体总的阳性预测值为76.3%。NIPT对于21-三体、18-三体、13-三体和性染色体非整倍体的阳性预测值分别为95.0%、88.8%、75.0%和53.3%。经CMA分析确认,在101例NIPT提示21-三体高风险病例中,以高龄筛查组阳性预测值最高(100.0%),3组间比较,差异有统计学意义(P=0.001)。在90例NIPT提示性染色体非整倍体高风险病例中,仍以高龄筛查组阳性预测值最高(61.9%),3组间比较,差异有统计学意义(P=0.002)。12例NIPT提示染色体非整倍体高风险经CMA检测存在CNV。结论:对于21-三体和性染色体非整倍体,不同人群中高龄组的NIPT阳性预测值最为显著,临床上对于此类人群,更要引起足够的重视,做好跟踪随访。CMA较传统的染色体核型分析能额外检出CNV,更能提高染色体结构异常的检出率。     

高龄孕妇胎儿基因组拷贝数变异发生率研究

目的探讨应用基因组拷贝数变异测序(CNV-seq)技术检测高龄孕妇羊水中基因组拷贝数变异(CNV)的发生率。方法收集2018年1月1日至12月31日因年龄高风险在哈尔滨医科大学附属第一医院应用羊膜腔穿刺术进行产前诊断的孕妇羊水标本247例。对胎儿羊水细胞进行G显带染色体核型分析的同时应用CNV-seq检测羊水细胞基因组DNA,统计在羊水染色体核型分析正常的高龄孕妇中CNV的发生率。结果 247例羊水标本中,染色体核型分析和CNV-seq同时检测出染色体非整倍体共计21例,其中21-三体综合征14例,18-三体综合征2例,13-三体综合征1例,性染色体非整倍体4例。染色体核型分析检出性染色体非整倍体1例(为45,XO[3]/46,XX[97]);染色体结构异常9例,其中染色体易位4例,染色体正常变异3例,2例染色体结构异常经CNV-seq检测最终确定异常片段的来源。染色体核型异常总检出率为12.55%(31/247)。在216例染色体核型结果正常的羊水标本中,CNV-seq额外检出明确致病CNV 4例,临床意义未明的染色体微缺失6例(其中1例为父源,1例为母源,其余4例未进行夫妇验证),CNV多态性15例。羊水样本中CNV的总检出率为19.43%(48/247),CNV-seq比染色体核型分析额外增加了6.88%的检出率,其中明确致病的CNV检出率为1.62%(4/247)。结论对进行有创产前诊断的高龄孕妇,在排除了常见的非整倍体异常后也应建议进行CNV检测。     

细菌人工染色体标记-微球鉴别/分离法对1973份羊水的快速产前诊断

目的探讨细菌人工染色体标记-微球鉴别/分离法(bacterial artificial chromosomes-on-beads,BoBs)在产前诊断中的应用价值。方法对2015年1月至2016年6月西京医院产前诊断中心1973例有产前诊断指征的孕妇羊水细胞进行BoBs检测和染色体核型分析。结果 1973例羊水中,共发现异常核型150例,其中染色体核型分析检出145例,BoBs检出133例。不同产前诊断指征下,无创DNA产前检测(NIPT)高风险组的染色体异常检出率最高,占该指征的81.82%。150例异常核型中,BoBs和核型分析均检出染色体非整倍体124例,占异常核型的82.67%;BoBs检出染色体微缺失/微重复综合征5例,核型分析结果均未见异常;染色体核型分析检出性染色体嵌合7例,BoBs检出4例;染色体核型分析检出结构异常14例,BoBs结果未见异常。结论 BoBs技术可以快速检测染色体非整倍体和基因微缺失/微重复,联合染色体核型、基因芯片(chromosomal microarray analysis,CMA)及荧光原位杂交技术(fluorescence in situ hybridization,FISH)可对染色体结构异常和低嵌合正确检出,大大提高了产前诊断的效率与准确性。     

Anonymous and non-anonymous gamete and embryo donations: pros and cons

Consequences of legal restrictions concerning gamete and embryo donation have been increasingly discussed during the last three decades, partly dependent upon the fact that assisted reproductive technologies have opened up new medico-technical possibilities. In the present communication we want to elucidate some of the problems and consequences of a Swedish law from 1985 requiring that all sperm donors should be non-anonymous. A similar law was passed in 2003 for oocyte donation. Such legal restrictions do not seem to be in the interest of the infertile couple who, to a large extent, seek treatment abroad where the laws are more liberal or non-restrictive (reproductive tourism). Whether or not the Swedish law is in the best interest for the offspring remains to be seen.     

Blood lead and cadmium and birth weight among sub-arctic and arctic populations of Norway and Russia

BACKGROUND: Delivering women and their newborns in the Kola Peninsula of Russia and the neighboring arctic area of Norway were studied to explore relationships between maternal cadmium and lead status and birth weight as a pregnancy outcome. METHODS: Life-style information, maternal blood and cord blood specimens were collected from 50 consecutive mother-infant pairs from hospital delivery departments in three Russian and three Norwegian communities. Pregnancy outcomes were verified by consulting medical records. Lead and cadmium were determined in the blood samples by electrothermal atomic absorption spectrometry. RESULTS: The median blood-cadmium concentration for the Russian mothers was 2.2 nmol/L (n = 148) versus 1.8 nmol/L in the Norwegian group (n = 114, p = 0.55). A weak association was observed between maternal cadmium and amount smoked (r = 0.30, p<0.001); no correlation was found between maternal blood cadmium and birth weight. The corresponding maternal lead values were 0.14 (Russia) and 0.06 micromol/L (Norway), p<0.001. The latter lead concentration constitutes one of the lowest adult population values reported to date. Maternal and cord blood lead levels were strongly correlated (r = 0.88, p<0.001). In a multivariate linear regression model, maternal blood lead was recognized as a negative explanatory variable (p<0.05) for birth weight and child's body mass index (BMIC), with or without adjustment for gestational age. A similar association was suggested by ANOVA-analysis of maternal blood lead by quartiles. CONCLUSION: Maternal blood-lead level as an environmental factor is an apparent predictor of low birth weight and BMIC. It reduced substantially the contribution of a country factor in explaining the observed differences in birth weight.     

Adiposity and Endometriosis Severity and Typology

Study ObjectivePrior research has collectively shown that endometriosis is inversely related to women's adiposity. The aim of this study was to assess whether this inverse relationship holds true by disease severity and typology.DesignCross-sectional study among women with no prior diagnosis of endometriosis.SettingFourteen clinical centers in Salt Lake City, UT, and San Francisco, CA.PatientsA total of 495 women (of which 473 were analyzed), aged 18–44 years, were enrolled in the operative cohort of the Endometriosis, Natural History, Diagnosis, and Outcomes (ENDO) Study.InterventionsGynecologic laparoscopy/laparotomy regardless of clinical indication.Measurements and Main ResultsParticipants underwent anthropometric assessments, body composition measurements, and evaluations of body fat distribution ratios before surgery. Surgeons completed a standardized operative report immediately after surgery to capture revised American Society for Reproductive Medicine staging (I–IV) and typology of disease (superficial endometriosis [SE], ovarian endometrioma [OE], and deep infiltrating endometriosis [DIE]). Linear mixed models, taking into account within-clinical-center correlation, were used to generate least square means (95% confidence intervals) to assess differences in adiposity measures by endometriosis stage (no endometriosis, I–IV) and typology (no endometriosis, SE, DIE, OE, OE + DIE) adjusting for age, race/ethnicity, and parity. Although most confidence intervals were wide and overlapping, 3 general impressions emerged: (1) women with incident endometriosis had the lowest anthropometric/body composition indicators compared with those without incident endometriosis, (2) women with stage I or IV endometriosis had lower indicators compared with women with stage II or III, and (3) women with OE and/or DIE tended to have the lowest indicators, whereas women with SE had the highest indicators.ConclusionOur research highlights that the relationship between women's adiposity and endometriosis severity and typology may be more complicated than prior research indicates.     

Toll样受体与子宫内膜及子宫内膜异位症

子宫内膜异位症(EMs)发病机制尚未完全阐明.大量研究表明,免疫因素在EMs的发病机制中起重要作用.EMs免疫应答异常主要是巨噬细胞数量和活性增加及其分泌产物,如生长因子、细胞因子和血管生成因子的改变.Toll样受体(TLRs)识别特异性的病原体相关分子模式,启动和介导免疫应答,在固有免疫中发挥重要作用,并诱导产生适应性免疫反应.TLRs在正常子宫内膜中的生理作用以及在EMs中的相关研究已逐步开展,对其深人认识和研究将为EMs诊断、治疗和预后判断提供新思路和手段.