运用胚外体腔液产前诊断α地中海贫血的临床研究

目的探讨胚外体腔穿刺液应用于α地中海贫血产前诊断的安全性及准确性。方法对40例妊娠6～10周、要求人工流产的单胎妊娠孕妇在终止妊娠前行胚外体腔穿刺术抽吸胚外体腔液，吸官术后取绒毛，采用聚合酶链反应(PCR)检测胚外体腔细胞及绒毛的α地中海贫血基因，比较两种标本诊断的符合率。结果36例胚外体腔细胞成功地扩增α地中海贫血基因，结果均与绒毛检测结果相符。结论通过胚外体腔穿刺术取胚外体腔细胞行PCR检测可用于α地中海贫血的早期产前诊断，具有可行性。     

胚外体腔穿刺术应用于α地中海贫血产前诊断的探讨

目的　探讨胚外体腔穿刺术应用于α地中海贫血产前诊断的可行性。方法　对 5 0例妊娠 6 10周、要求人工流产的单胎妊娠孕妇在终止妊娠前行胚外体腔穿刺术抽吸胚外体腔液 ,吸宫术后取绒毛 ,采用聚合酶链反应(PCR)扩增胚外体腔细胞及绒毛的α地中海贫血基因 ,比较两种标本诊断的符合率。结果　 40例胚外体腔细胞成功地扩增α地中海贫血基因 ,结果均与绒毛相符。结论　通过胚外体腔穿刺术取胚外体腔细胞行PCR检测可用于α地中海贫血的早期产前诊断 ,但应进一步提高诊断方法的敏感性。     

胚外体腔液中胎儿细胞DNA检测在产前诊断中的应用

目的探讨胚外体腔液中细胞DNA检测用于孕早期诊断性连锁遗传疾病的可行性和准确性。方法对25例孕6～11周、要求人工流产的孕妇术前行胚外体腔穿刺术,抽取胚外体腔液0．5～3ml。采用煮沸法对胚外体腔液进行DNA抽提,Picogreen染料对DNA进行定量分析。采用X,Y两对引物对DNA进行PCR检测。随后行人工流产术,取少量绒毛组织进行染色体核型分析,并与胚外体腔液中的DNA检测结果作对照。结果0．5～3ml胚外体腔液中含有750—6270个胎儿细胞。男性胎儿DNA扩增结果出现两个条带（X,Y）11例,女性胎儿出现1个条带（X）14例。与绒毛组织染色体核型分析结果完全一致。结论胚外体腔液中细胞DNA检测,用于孕早期性连锁遗传疾病的产前诊断具有快速、敏感的特点。     

胚外体腔穿刺术应用于α地中海盆血产前诊断的探讨

目的：探讨胚外体腔穿刺术应用于α地中海贫血产前诊断的可行性。方法：对50例妊娠6－10周、要求人工流产的单胎妊娠孕妇在终止妊娠前行胚外体腔穿刺术抽吸胚外体腔液，吸宫术后取绒毛，采用聚合酶链反应（PCR）扩增胚外体腔细胞及绒毛的α地中海贫血基因，比较两种标本诊断的符合率。结果：40例胚外体腔细胞成功地扩增α地中海贫血基因，结果均与绒毛相符。结论：通过胚外体腔穿刺术取胚外体腔细胞行PCR检测可用于α地中海贫血的早期产前诊断，但应进一步提高诊断方法的敏感性。     

早孕胚胎胚外环境的临床研究

目的:研究早孕胚胎的胚外环境。方法:非意愿妊娠人工流产前在B超下对28例孕6～11周胚胎进行胚外体腔穿刺,抽取体腔液,同时抽取孕妇静脉血,其中8例同时穿刺羊膜腔抽取羊膜腔液,行生化检测。结果:母血中碱性磷酸酶(AKP)、总蛋白远高于胚外体腔液及羊膜腔液(P<0.01);Na~+、Cl~+、肌酐浓度也较高(P<0.05),K~+、Ca~(2+)浓度基本相似(P>0.05);葡萄糖浓度略高于体腔液(P<0.01),与羊膜腔液基本相似(P>0.05)。胚外体腔液蛋白浓度是羊膜腔液的3倍,Ca~(2+)及肌酐浓度也较高(P<0.05)。其余指标无显著性差异。结论:早孕阶段母体-绒毛间隙-羊膜囊存在交流,并呈现一定梯度,绒毛膜及羊膜起了半透膜的作用,限制营养物质交换和代谢,从而保持胚胎在早期发育中有相对稳定的内环境。     

B超引导下的脐静脉穿刺术在产前诊断应用中的安全性研究

目的探讨B超引导下的脐静脉穿刺术在产前诊断中的成功率和安全性。方法回顾性分析1990年3月～2003年6月对2403例因各种原因在B超引导下的脐静脉穿刺术进行产前诊断的孕妇临床资料,观察穿刺手术的成功率和并发症。结果一次性穿刺成功为2368例(98．5％),二次穿刺成功为2384例(99．2%)。75．5％的孕妇可在5min内完成穿刺术,93．0％的孕妇在10min内完成穿刺术。手术并发症包括：脐带或胎盘渗血315例(13．1％),胎儿心动过缓125例(5．2％),一次性穿刺失败35例(1．5％),流产18例(0．8％),早产4例(0．2e%),绒毛膜羊膜炎2例(0．1％)。结论脐静脉穿刺术是一项较为安全而简单易行的产前诊断取材技术。     

胎儿躯干周径:一个新的生物统计学参数

作者对131例无妊娠合并症、单胎、孕7～12周的孕妇用超声波检查胎儿心脏活动、胎儿顶-臀长度(CRL)和躯干周径(TC),发现CRL和TC两者均与孕龄成曲线相关,其关系是: 孕龄=5.23+(2.53)CRL+(-0.345)CRL~2+(0.0172)CRL(?) 孕龄=6.23+(0.16)TC+(-0.0011)TC~2 TC=2平均躯干直径×3.14 躯干直径为在胎儿心脏搏动点尾端垂直于躯干长轴测得的躯干直径。作者认为妊娠早期用测量胎儿长轴或CRL来     

不同标本来源对产前筛查结果影响的分析

目的:探讨不同标本来源对孕中期各筛查指标中位数倍数值(MoM)中位数(mMoM)及筛查效率 的影响。 方法:以广西壮族自治区妇幼保健院遗传代谢中心实验室2018 年1 月至2019 年5 月进行孕中期 唐氏综合征筛查的10318 例孕妇作为研究对象,其中院内采集样本 6855 例,合作单位送检样本 3463 例。 按照不同来源及孕周计算方式,分为院内顶臀长(CRL) / 末次月经(LMP)组(F1CRL/ F1LMP 组,4768 例), 院内末次月经组(F2LMP 组,2087 例),送检单位末次月经组(F3LMP 组,3463 例),通过 F1CRL/ F1LMP 组 建立本地标准化中位数方程,分别加入同期 F2LMP 组、F3LMP 组数据进行整体分析。 采用折线图、累积和 图对各筛查指标[甲胎蛋白(AFP)、游离 β-人绒毛膜促性腺激素(f-β-hCG)和游离雌三醇(uE3 )]mMoM 值 进行对比,并随访分析其筛查效率。 结果:①F1CRL、F1LMP 两方程各筛查指标 mMoM 值均介于0. 9 ~1. 1, 折线图、累积和图变化趋势无明显变化,经随访妊娠结局确诊唐氏综合征 0 例,F1CRL 的筛查阳性率高于 F1LMP(2. 33% vs 3. 25%,P <0. 05);②加入同期 F2LMP 组、F3LMP 组的数据示:AFP mMoM 值的影响均不 明显,而 F3LMP 组中 f-β-hCG mMoM 值升高、uE3 mMoM 值降低的趋势较为明显,F2LMP 组变化趋势不明 显,经随访妊娠结局 F2LMP 组、F3LMP 组分别确诊21-三体2 例、3 例,两组数据的筛查阳性率比较,差异有 统计学意义(5. 37% vs 7. 28%,P <0. 05),且 F2LMP 组阳性预测值(1/ 56)略高于 F3LMP 组(1/ 84)。 结论: CRL 与 LMP 两种不同孕周计算方式对筛查指标 mMoM 值影响较小,但 CRL 计算孕周应用于孕中期唐氏 综合征筛查的效率优于 LMP;筛查指标 f-β-hCG、uE3 mMoM 值受标本来源的影响较明显,故混合型标本来 源的数据不宜共用 f-β-hCG 及 uE3 的中位数方程进行风险结果评估。     

妊娠中期单一的母血β-HCG水平下降与不良的妊娠结局无相关性

本研究的目的是探讨母血筛查中的低水平HCG是否导致不良的妊娠结局。 1999年 6月～ 2 0 0 1年 11月期间对孕 15～ 2 0周行母血筛查的妇女进行研究。以血AFP >0 .5、<2 .0中位数的倍数 (MoM) ,血β HCG >0 .5、<2 .0MoM ,雌三醇 (E3 ) >0 .6、<2 .0MoM为正常。选择母血 β HCG≤ 0 .5MoM ,AFP、E3 值正常者 14 6例作为研究病例 ,同时随机选择AFP、E3 、血 β HCG值均正常的妇女 2 92例作对照分析。两组孕妇的年龄、种族、以往不良妊娠结局、母血筛查时的胎龄及吸烟状况均无差异。研究组的母血 β HCG值为0 .4± 0 .1MoM ,显著…     

胚外体腔穿刺的现状

胚外体腔穿刺是于孕６～１０周在Ｂ超引导下,用穿刺针刺入胚外体腔抽吸液体作产前诊断,并可注入异体移植物行宫内治疗或诱导免疫耐受的一种新技术。它起源于１９９１年,由Ｗａｔｈｅｎ等［１］首次描述并操作。１９９３年由Ｊｕｒｋｏｖｉｃ等［２］首次应用于产前诊断...     

Aneuploidy screening in coelomic samples using fluorescence in situ hybridisation (FISH)

OBJECTIVES: Coelocentesis is the earliest invasive prenatal diagnostic procedure that has recently been used in ongoing pregnancies to identify single gene defects. Aneuploidy screening has not yet been performed in ongoing pregnancies following coelocentesis, but experimental studies have demonstrated the ability of determining the copy number of chromosomes 13, 18, 21, X and Y in uncultured coelomic samples, by FISH or PCR. The aim of this study was to extend previous studies and investigate the feasibility of analysing uncultured coelomic fluid samples for 11 chromosomes using fluorescence in situ hybridisation (FISH). METHODS: Coelocentesis was performed in 12 singleton pregnancies at 6 to 9 weeks of gestation immediately before surgical termination of pregnancy. Fluorescence probes for chromosomes 3,7,9,13,16,17,18,21,22, X and Y were applied on uncultured coelomic-fluid samples and placental tissue. In cases where coelomic cells were not of a sufficient amount, chromosomes X and Y were analysed in a second layer of hybridisation. RESULTS: Successful analysis by FISH was possible in all cases and the results from the coelomic fluid were concordant with those from the analysis of placental tissue and obtained within a few hours of receiving the samples. Problems associated with limited cell numbers were overcome by the application of a second layer of FISH. This sequential approach has also enabled accurate identification of maternal-cell contamination in male samples. CONCLUSION: Analysis of 11 chromosomes using FISH in coelomic fluid samples is feasible and it has the potential to be applied for rapid aneuploidy screening, should coelocentesis be used clinically as an early, invasive prenatal diagnostic tool.     

Studies on features of fetal movement and development of human fetus with use of fetal actogram

In 167 normal fetuses at 26 to 41 weeks of gestation, features of fetal movement and fetal development were investigated with use of actocardiograph in connection with a microcomputer system. The signals of fetal movement obtained by actocardiograph were stored in a floppy disc every 250 ms for 5 minutes through an AD-converter, and were analyzed every 5 minutes with the computer to reveal 3-dimensional (3-D) histograms. The 3-D histogram of fetal movement was composed of number, amplitude and interval of the signals in 11 voltage steps between 0.05 and 0.55V. The histogram clearly indicated state of fetal behavior, being either resting or active state. Fetal movement such as rolling movement, breathing movement and hiccup could be also identified with the computer analysis. In 68 normal fetuses at 14 to 41 weeks of gestation, the cross-correlation between fetal movement and fetal heart rate (FHR) were examined with the computer analysis. Finally fetal responses to acoustic and light stimulation were evaluated with use of pure-tone generator and flashlight. Acoustic stimulation was carried out in 53 normal fetuses at 28 to 41 weeks and light stimulation was performed in 116 normal fetuses at 18 to 41 weeks of gestation. The fetal responses were evaluated with actocardiogram. As a result, 1) Frequency in active state decreased and resting state increased as gestational weeks advanced, and then the frequencies of both state remained constant after 37 weeks of gestation. Duration of resting state also increased from 26 weeks to 37 weeks. These observations may suggest that fetal behavior can be established by 37 weeks of gestation. 2) Frequency in rolling movement decreased until 37 weeks of gestation, and then the movement increased during 38-41 weeks. Frequency in breathing movement increased to 33 weeks of gestation, then it remained constant. Hiccup occurred most frequently at 30-33 weeks, and it decreased thereafter. The function in fetal respiratory movement may be accomplished by 33 weeks of gestation. 3) Positive cross-correlation between fetal movement and FHR was observed as gestational weeks advanced. The correlation coefficient increased from 14 weeks up to 41 weeks. Acceleration of FHR following fetal movement eventually occurred in fetuses at 16 weeks, but the onset of acceleration delayed than normally occurred in developed fetuses. The delay was shortened in fetuses at 24 weeks and this was comparable to the delay in developed fetuses. These results suggest that the linkage of the acceleration of FHR with fetal movement is established at 24 weeks of gestation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Early amniocentesis: alphafetoprotein levels in amniotic fluid, extraembryonic coelomic fluid and maternal serum between 8 and 13 weeks.

OBJECTIVE--The aim was to establish a normal range of alphafetoprotein (AFP) concentrations in amniotic fluid from 8 to 12 weeks gestation, and to determine any difference between AFP levels in amniotic fluid and extraembryonic coelomic fluid. DESIGN AND SUBJECTS--150 women had a transvaginal ultrasound guided amniocentesis before termination of an apparently normal first trimester pregnancy. Separately identified samples of amniotic fluid and extraembryonic coelomic fluid were obtained and assayed by radioimmunoassay for AFP. RESULTS--In amniotic fluid, very high levels of AFP were present at 8 weeks, levels falling rapidly up to 10 weeks after which there was a slight rise. Thus over the period 8 to 10 weeks, there was a significant inverse correlation between amniotic fluid AFP and gestational age (r = 0.67; P less than 0.001). In extraembryonic coelomic fluid, by contrast there was no trend in AFP relative to gestational age. CONCLUSIONS--The rapidly changing levels of AFP from 8 to 10 weeks as well as the small volume of the amniotic cavity makes the use of amniocentesis impracticable before 11 weeks gestation. The lack of any relation between AFP levels in amniotic fluid and extraembryonic coelomic fluid emphasises the importance of identifying the site of amniocentesis in the first trimester.     

Prenatal paternity testing using DNA extracted from coelomic cells

OBJECTIVE: Prenatal paternity testing can be performed following invasive prenatal diagnosis with amniocentesis or CVS. Coelocentesis is a new technique that could be used as an alternative method early in the first trimester of pregnancy. The aim of this study is to investigate the potential use of the DNA extracted from coelomic cells in the prenatal paternity testing. METHODS: Coelocentesis was performed in 20 singleton pregnancies at 7-9 weeks of gestation immediately before surgical termination of pregnancy. Chorionic cells from the placenta and blood cells from the parents were processed by the standard salt extraction method. Two loci, TPO and Apo B, were used for paternity testing in the DNA of coelomic cells, chorionic cells and blood cells. RESULTS: There was concordance in the results obtained from the coelomic cells and chorionic villi. In two cases only the polymorphisms used were not conclusively informative for paternity exclusion. CONCLUSIONS: Coelomic cells are potentially useful for early paternity testing.     

Evaluation of fetal heart rate variation during amniocentesis: correlation with fetal karyotype

Objective.?We monitored the fetal heart rate (FHR) during amniocentesis in fetuses at 16–18 weeks of gestation and investigated whether an abnormal FHR is associated with chromosomal abnormalities.

Methods.?This prospective study involves 807 women at 16–18 weeks of gestation who underwent genetic amniocentesis. The FHR, expressed as beats for minute, is recorded before (FHR1), immediately after (FHR2) and 60?min after (FHR3) the invasive procedure. Structural malformations detected by ultrasound and multiple pregnancy are excluded from the study.

Results.?Chromosomal abnormalities have been diagnosed in 27 fetuses. A mean FHR decrease after amniocentesis has been observed in normal and in abnormal fetuses. The mean variation during amniocentesis is significant in both groups (P?<?0.01). The comparison between the mean FHR of the two groups shows no differences in FHR1 and FHR2 (P?>?0.05) but a significant difference in FHR3 (P?<?0.05).

Conclusion.?The FHR decreases after amniocentesis; the decrease is larger in chromosomally abnormal fetuses than in normal fetuses. This difference in heart rate reaction to amniocentesis might be due to cardiac defects or developmental delay associated with the abnormal karyotype.     

Transabdominal coelocentesis as early source of fetal DNA for chromosomal and molecular diagnosis

Abstract

This study reports a comparative analysis between results of transabdominal coelocentesis and traditional invasive procedure in order to assess the usefulness of coelocentesis as a source of fetal DNA for molecular and chromosomal analysis. A number of 28 women were included in the study. A successful sampling of coelomic fluid was obtained in 25 women by transabdominal procedure. A positive amplification of DNA with QF-PCR techniques was obtained in 90% of cases, while 10% of cases failed to reveal interpretable results. Although all samples were cultured, the growth rate was not sufficient to determine karyotypes within 2 weeks. Five samples were selected to be analyzed by array-based comparative genomic hybridization (a-CGH) but the interpretation of these results was difficult and ambiguous. Our results suggest that transabdominal coelocentesis is suitable for the detection of single DNA variation and for QF-PCR analysis, while further experiments are needed to develop optimized protocols for traditional karyotyping and array-analysis.     

The clustering of fetal heart rate changes and fetal movements in pregnancies between 20 and 30 weeks of gestation

The temporal organization (nonrandomness) of fetal heart rate (FHR), fetal movement, and their association was evaluated in 20 normal pregnancies at 20 to 22 or 28 to 30 weeks of gestation with the use of external electronic fetal monitoring and statistical analysis with the runs test. At 20 to 22 weeks, significant clustering was found in one of 10 pregnancies for FHR change, two of 10 pregnancies for fetal movement, and one of 10 pregnancies for FHR changes associated with fetal movement. At 28 to 30 weeks, significant clustering was found in eight of 10 pregnancies for FHR change, eight of 10 pregnancies for fetal movement, and uli 10 pregnances for FHR change associated with fetal movement. There was significantly more clustering of FHR change, fetal movement, and FHR change associated with fetal movement at 28 to 30 weeks of gestation than at 20 to 22 weeks of gestation. Clustering of FHR changes was highly correlated with clustering of fetal movement.     

Chorionic villus sampling significantly affects fetal cardiovascular function.

OBJECTIVE: To assess the effects of chorionic villus sampling (CVS) on fetal heart rate (FHR). METHODS: A prospective longitudinal study was conducted among 300 patients undergoing transabdominal CVS between 8 and 13 weeks of gestation. Duration of the procedure, number of needle passes, sample weight, maternal age, fetal gender, and FHR response to CVS were recorded. RESULTS: The FHR before but not after CVS was inversely correlated with gestational age (r = -0.406, p < 0.001). Conversely, following CVS, no correlation was observed between FHR and gestational age (r = -0.06, p = 0.27). The difference between FHR after CVS and that obtained before CVS (delta FHR) increased with increasing gestational age at sampling (r = 0.372, p < 0.0001), decreased with increasing specimen weight (r = -0.16, p = 0.01) and increased with increasing maternal age (r = 0.22, p < 0.0001). Duration of the procedure, fetal gender and number of needle passes did not affect delta FHR. Multiple logistic regression indicated that gestational age at CVS and maternal age but not the other variables significantly affected delta FHR and together they accounted for over 22% of the variance (R(2) = 0.224, p < 0.0001). CONCLUSIONS: In summary, our results suggest that acute fetal hemodynamic changes accompany CVS and that these changes vary with gestational age.     

Computerised cardiotocography following vibro-acoustic stimulation.

To study the effect of vibro-acoustic stimulation (VAS) to the mean fetal heart rate (FHR), period of high and low FHR variation, overall variation (msec and bpm) and short term variation (msec). In a prospective study 17 pregnant women between 34-42 weeks gestation admitted to antenatal ward for obstetric complications two 60 min FHR recording was carried out with an interval of 30 min in between recordings. On a random basis the fetus was stimulated by a vibro-acoustic stimulator for 5 sec at the beginning of one of the two 60 min FHR recordings. Automated analysis of the FHR, tocodynamometry, and maternal perception of fetal movements was done by a commercially available computerized programme (System 8000). It was possible to obtain the two 60 min recordings with signal loss of < 10% in 12 out of 17 patients. No changes were observed when the FHR parameters for 60 min after VAS was compared with the control period. When analyzed in segments of 0 to 10, 11 to 20, 20 to 40 and 41 to 60 min the mean baseline FHR was significantly higher after VAS during the first 10 minutes compared with any 10 or 20 min segment of the control period or any such segments 10 min after VAS. Concomitantly the overall variation and short-term variation was significantly lower during the first 10 minutes following VAS (p < 0.05) compared with the parameters in the corresponding periods during the control period.(ABSTRACT TRUNCATED AT 250 WORDS)