运用多普勒超声心动图技术对染色体三体异常小鼠胚胎心脏…

对２０个１６三体染色体异常小鼠胚胎进行了多普勒超声心动图研究。龄从１０天至１５天。９例发现流入道返流波型，６例发现流出道反流波型。１２０例染色体正常胚胎无一例发现返流波型。１６三体小鼠模型作为先天性心脏病研究的理想模型，本文首次提供了对此类模型胚胎血液动力学研究的非创伤性监测方法。     

超声波抗早孕的动物实验研究Ⅲ.超声波对妊娠第7天小鼠的…

本文报道了一种应用治疗剂量超声抗鼠早孕的方法。结果显示：用１２Ｗ／ｃｍ＾２、１６Ｗ／ｃｍ＾２、２０Ｗ／ｃｍ＾２三组声强辐照小鼠妊娠第７天在体胚胎着床点的组织学急性改变为胎盘出血，电镜下见胎盘和胚胎细胞多数受到严重损伤和细胞质膜失去连续性、细胞核碎裂等；子宫内膜和平滑肌细胞出现线粒体肿胀、内膜细胞的胞质内有的局灶性坏死。受辐照小鼠继续妊娠至第１６天时处死发现受辐照的胚胎１００％死亡，光镜下见胎盘、胚     

多重连接探针扩增技术在胚胎停止发育染色体非整倍体异常分析中的应用

目的：探讨多重连接探针扩增（multiplex ligation-dependent probe amplification,MLPA）技术在分析胚胎停止发育（以下简称胚胎停育）流产绒毛组织染色体非整倍体异常中的应用,探讨染色体非整倍体异常导致胚胎停育与胎儿性别、患者年龄间的关系。方法：收集324例临床诊断为胚胎停育患者的流产绒毛组织,采用MLPA技术进行染色体检测,并分析检测结果与胎儿性别及患者年龄间的相关性。结果：在324例胚胎停育患者中,检出绒毛染色体非整倍体异常共68例（20.99%）,其中常染色体三体33例（13-三体12例、18-三体6例、21-三体15例）,常染色体缺失3例（13-单体1例、21-单体2例）,性染色体数目异常32例（45-XO 15例、47-XXY 13例、47-XXX 1例、47-XYY 3例）。常染色体数目异常组与染色体数目正常组间的胎儿性别分布差异无统计学意义（P>0.05）。对绒毛染色体数目正常组[平均年龄（32±6）岁]、常染色体数目异常组[平均年龄（34±7）岁]、性染色体数目异常组[平均年龄（28±6）岁] 3组患者的年龄进行比较,发现差异有统计学意义（P<0.05）。结论：MLPA技术可以作为一种辅助检测手段,用于检测胚胎停育流产绒毛组织染色体的非整倍体异常,染色体数目异常导致的胚胎停育与胎儿的性别无关,但与患者的年龄有关,胚胎常染色体数目异常组的孕妇年龄大于染色体数目正常组及性染色体数目异常组。     

染色体核型分析与荧光原位杂交用于胚胎停育患者病因学诊断

目的 探讨染色体核型分析技术及荧光原位杂交（FISH）技术两种检测方法在胚胎停育患者胎儿病因学诊断中的应用。 方法 对78例胚胎停育患者清宫后的妊娠组织行细胞培养后做染色体核型分析;若培养失败,采用FISH进行分析。 结果 78例胚胎停育患者的妊娠组织,染色体培养且成功用于核型分析者38例,其中检出正常核型23例、异常核型15例、常染色体三体8例、三倍体1例、性染色体异常3例、结构异常3例。40例染色体培养失败,采取FISH检测,检出正常染色体15例,异常染色体25例,其中常染色体三体占19例、三倍体2例、45XO 4例。 结论 胚胎停育的主要原因是胚胎染色体异常导致。通过染色体核型分析及FISH检测对胚胎停育的妊娠组织进行分析,两种方法对异常核型和常染色体数目异常的检出有明显差异,对胚胎坏死时间长的妊娠组织,FISH成功率更高。临床上应结合两种检测方法用于胚胎停育的原因分析,为下次妊娠提出指导意见。       

应用荧光原位杂交技术检测54例胚胎停育患者绒毛染色体数目异常的分析

目的应用荧光原位杂交(fluorescence in situ hybridization,FISH)方法对胚胎停育的绒毛组织进行分析,明确胚胎停育与染色体数目异常的情况。方法选择针对13、16、18、21、22、X、Y染色体特异性荧光探针,应用FISH方法,对154例胚胎停育的绒毛组织中的染色体数目进行分析,并了解胚胎停育患者绒毛染色体数目异常情况及其与患者年龄及流产次数的差异。结果 1154例胚胎停育患者染色体数目异常56例(36.4%),其中,常染色体三体40例(71.4%),多倍体4例(7.1%),45,XO 12例(21.4%)。2绒毛染色体数目异常患者的年龄[(32.14±5.21)岁]与染色体数目正常患者的年龄[(29.49±4.07)岁]比较,差异有统计学意义(u=2.145,P<0.05)。3流产次数2次及以上的染色体数目异常发生率(44.8%)高于流产次数为1次的发生率(25.4%),差异有统计学意义(χ2=6.19,P<0.05)。结论染色体数目异常是胚胎停育的重要发病因素,其中常染色体三体最多见;对2次及以上的流产患者,更应关注是否有染色体异常的发生。     

扬州地区58例21-三体综合征儿童细胞遗传学分析

目的探讨扬州地区21-三体综合征儿童染色体核型分布及特点。方法对2006年11月至2015年12月前来该院门诊进行遗传咨询的433例儿童进行外周血染色体核型分析。采用外周血淋巴细胞培养和G显带技术,进行细胞遗传学分析。结果共发现21-三体综合征58例。其中单纯型21-三体占93.10%(54/58);嵌合型占3.45%(2/58);易位型占3.45%(2/58)。在单纯型21-三体患儿的性别比(男∶女)为1∶1.16。结论扬州地区约93%的21-三体综合征患者染色体核型是单纯型21-三体,男孩和女孩21-三体综合征患儿比例相当。     

孕妇外周血游离 ＤＮＡ 检测用于胎儿 １６－三体筛查的 临床价值

摘要：目的 探讨孕妇血浆游离 ＤＮＡ（ｃｆ ＤＮＡ）检测技术用于胎儿 １６－三体综合征（１６－三体）筛查的临床应用价值。方法 收 集２０１７ 年 １ 月至 ２０２１ 年 １２ 月在南京市妇幼保健院进行孕妇血浆 ｃｆ ＤＮＡ 检测且结果为 １６－三体高风险的孕妇，对其侵入性 产前诊断验证结果，并对妊娠结局进行回顾性分析。结果 在 ５６ ２４７ 例进行 ｃｆ ＤＮＡ 检测的孕妇中，检出 １６－三体高风险 ２２ 例（０．０４％，２２ ／ ５６ ２４７）；其中 ８ 例选择了侵入性产前诊断，检出 ２ 例单亲二倍体，未检出胎儿真阳性，阳性预期值为 ０；妊娠结 局回访发现 ２２ 例 １６－三体高风险孕妇中，１９ 例（８７．４％，１９ ／ ２２）孕妇存在流产、胎儿发育异常、早产、生长发育迟缓等。结论 血浆 ｃｆ ＤＮＡ 检测提示１６－三体高风险中胎儿真阳性的可能性小，多数可能是限制性胎盘嵌合；ｃｆＤＮＡ 检出 １６－三体高风险中 大部分孕妇存在不良妊娠结局，应密切关注这类胎儿的生长发育情况及围产期可能的并发症。     

低深度高通量测序技术在早期流产中的应用

目的:探讨低深度高通量测序技术(CNV-seq)在早期流产中的应用。方法:将2017年1月～2018年1月我院收治的265例早期流产患者(99例染色体核型异常)作为研究对象,取其绒毛或胚胎组织,提取DNA,采用低深度高通量测序技术分析其基因组信息,并与常规染色体核型相对照,分析染色体异常的类型及检出率情况。结果:265例早期流产患者中出现99例染色体核型异常者,发生率为37.36%;其中染色体异常核型分布:16三体12例(12.12%)、13三体6例(6.06%)、21三体9例(9.09%)、2三体3例(3.03%)、3三体4例(4.04%)、8三体3例(3.03%)、10三体4例(4.04%)、22三体3例(3.03%)、常染色体缺失或重排27例(27.27%)、45,X 21例(21.21%)、47,XYY 3例(3.03%)、47,XXX 4例(4.04%)。低深度高通量测序对核型异常检出率为100.00%,显著高于常规染色体分析的74.75%,比较具有统计学差异(P0.05)。结论:低深度高通量测序技术较常规染色体分析对早期流产患者基因组信息的检出率较高,值得临床选择。     

胚胎因子对小鼠红细胞免疫粘附功能的影响

本文采用补体致敏酵母菌血凝法，检测了胚胎因子对小鼠红细胞免疫粘附功能的影响，结果显示，对照组小鼠红细胞血凝阳性滴度多为１：４，实验组小鼠红细胞血凝阳性滴度多为１：８或１：１６。在血凝滴度１：８时，二者阳性率有明显差异（Ｐ〈０．０５），揭示胚胎因子对小鼠红细胞免疫粘附功能有增强作用。     

遗传学检验与I临床

四川地区重型β-地中海贫血患儿及双亲基因突变的研究，精子发生障碍患者Y染色体AZF微缺失的筛查及意义,无精子症和严重少精子症134例遗传学分析，16例多发性骨髓瘤患者的染色体分析,马凡综合征FBN1基因单倍型连锁分析,应用引物原位标记技术快速检测18三体综合征,短串联重复序列聚合酶链反应产前诊断多胎妊娠合子性质及常见染色体三体,染色体分析对恶性胸水的诊断价值     

Longitudinal in Utero Analysis of Engrailed-1 Knockout Mouse Embryonic Phenotypes Using High-Frequency Ultrasound

Large-scale international efforts to generate and analyze loss-of-function mutations in each of the approximately 20,000 protein-encoding gene mutations are ongoing using the “knockout” mouse as a model organism. Because one-third of gene knockouts are expected to result in embryonic lethality, it is important to develop non-invasive in utero imaging methods to detect and monitor mutant phenotypes in mouse embryos. We describe the utility of 3-D high-frequency (40-MHz) ultrasound (HFU) for longitudinal in utero imaging of mouse embryos between embryonic days (E) 11.5 and E14.5, which represent critical stages of brain and organ development. Engrailed-1 knockout (En1-ko) mouse embryos and their normal control littermates were imaged with HFU in 3-D, enabling visualization of morphological phenotypes in the developing brains, limbs and heads of the En1-ko embryos. Recently developed deep learning approaches were used to automatically segment the embryonic brain ventricles and bodies from the 3-D HFU images, allowing quantitative volumetric analyses of the En1-ko brain phenotypes. Taken together, these results show great promise for the application of longitudinal 3-D HFU to analyze knockout mouse embryos in utero.     

Evaluation of hypertrophic cardiomyopathy by Doppler color flow imaging: initial observations

We evaluated hypertrophic cardiomyopathy in 12 patients by Doppler color flow imaging and continuous-wave Doppler echocardiography. Mitral regurgitation was detected by continuous-wave Doppler echocardiography in eight patients and was related to the degree of systolic anterior motion of the mitral valve. Adequate color flow images were obtained in 10 of the 12 patients, and mitral regurgitation was demonstrated in 6. A qualitative and quantitative analysis of the color flow imaging revealed a temporal pattern in the left ventricular outflow tract that consisted of normal-velocity laminar flow during early systole followed by turbulent flow in midsystole. The maximal amount of mitral regurgitation on color flow imaging occurred late in systole, after the appearance of turbulent flow in the left ventricular outflow tract. Of the 12 patients, 10 had late-peaking continuous-wave Doppler velocity profiles in the left ventricular outflow tract. The peak velocity detected in the left ventricular outflow tract was positively correlated with the degree of systolic anterior motion of the mitral valve. Patients with higher peak velocities in the left ventricular outflow tract had prolonged ejection times. These findings on Doppler echocardiography support the concept of left ventricular outflow obstruction in some patients with hypertrophic cardiomyopathy.     

High-Throughput,High-Frequency 3-D Ultrasound for in Utero Analysis of Embryonic Mouse Brain Development

With the emergence of the mouse as the predominant model system for studying mammalian brain development, in utero imaging methods are urgently required to analyze the dynamics of brain growth and patterning in mouse embryos. To address this need, we combined synthetic focusing with a high-frequency (38-MHz) annular-array ultrasound imaging system for extended depth-of-field, coded excitation for improved penetration and respiratory-gated transmit/receive. This combination allowed non-invasive in utero acquisition of motion-free 3-D data from individual embryos in approximately 2 min, and data from four or more embryos in a pregnant mouse in less than 30 min. Data were acquired from 148 embryos spanning 5 d of early to mid-gestational stages of brain development. The results indicated that brain anatomy and cerebral vasculature can be imaged with this system and that quantitative analyses of segmented cerebral ventricles can be used to characterize volumetric changes associated with mouse brain development.     

Comparison of several noninvasive methods for estimation of pulmonary artery pressure

Noninvasive estimation of pulmonary artery pressure is an important component of cardiac ultrasound studies. A number of methods are available for estimation of pulmonary pressure, each with varying degrees of reported accuracy. To assess feasibility and accuracy, noninvasive pulmonary artery pressure estimates were performed in infants and children at the time of catheterization. Patients were examined prospectively until there were 50 patients, in whom each of six methods for estimation of pulmonary pressure had been accomplished. All patients had tricuspid and pulmonary regurgitation of less than severe degree and no structural, flow, or electrocardiographic abnormality known to compromise the six methods. Systolic pressure was estimated by the Burstin method and also from peak tricuspid regurgitation velocity. Mean pressure was estimated by acceleration time divided by ejection time from waveforms obtained from the right ventricular outflow tract and main pulmonary artery. Diastolic pressure was estimated by systolic time intervals and from end-diastolic pulmonary regurgitation velocity. Noninvasive estimates were compared with simultaneous or nearly simultaneous catheterization measurements. For systolic pressure Burstin estimates were accomplished in 89% with high accuracy (r = 0.97). Tricuspid regurgitation velocities were recorded in 82%, also with high accuracy (r = 0.96). Waveforms for mean pressure estimation were recorded in 98% to 100% of patients. Those from the right ventricular outflow tract corresponded well with catheterization pressures (r = 0.94), whereas those recorded from the main pulmonary artery offered poor prediction of pulmonary pressure (r = 0.63). Systolic time interval measurements were accomplished in only 65% and did not correlate highly with catheterization (r = 0.84). Diastolic pressure estimates based on pulmonary regurgitation velocity were recorded in 98% of subjects with high accuracy (r = 0.96). Each method had advantages and disadvantages. The Burstin method was accurate but technically demanding and is reported to be limited by heart rate and significant right-sided regurgitation. Peak tricuspid velocities proved unexpectedly difficult to record in some patients but when successful, provided excellent prediction of pressure. Recording of waveforms for ratios of acceleration time to ejection time proved easy, but accuracy was high only for outflow tract waveforms. Peculiarities of main pulmonary artery flow may have led to poor accuracy for ratios measured from that site. For diastolic pressure estimation, systolic time interval records were the most difficult to obtain and did not provide useful accuracy. In contrast, pulmonary regurgitation velocities were easily obtained and provided high accuracy results. This is a selected pediatric series, evaluating methods in nearly ideal circumstances.(ABSTRACT TRUNCATED AT 400 WORDS)     

Disruption of ECE-1 and ECE-2 reveals a role for endothelin-converting enzyme-2 in murine cardiac development

Endothelin-converting enzyme-1 and -2 (ECE-1 and -2) are membrane-bound metalloproteases that can cleave biologically the inactive endothelin-1 (ET-1) precursor to form active ET-1 in vitro. We previously reported developmental defects in specific subsets of neural crest-derived tissues, including branchial arch-derived craniofacial structures, aortic arch arteries, and the cardiac outflow tract in ECE-1 knockout mice. To examine the role of ECE-2 in cardiovascular development, we have now generated a null mutation in ECE-2 by homologous recombination. ECE-2 null mice develop normally, are healthy into adulthood, are fertile in both sexes, and live a normal life span. However, when they are bred into an ECE-1-null background, defects in cardiac outflow structures become more severe than those in ECE-1 single knockout embryos. In addition, ECE-1(-/-); ECE-2(-/-) double null embryos exhibited abnormal atrioventricular valve formation, a phenotype never seen in ECE-1 single knockout embryos. In the developing mouse heart, ECE-2 mRNA is expressed in the endocardial cushion mesenchyme from embyronic day (E) 12.5, in contrast to the endocardial expression of ECE-1. Levels of mature ET-1 and ET-2 in whole ECE-1(-/-); ECE-2(-/-) embryos at E12.5 do not differ appreciably from those of ECE-1(-/-) embryos. The significant residual ET-1/ET-2 in the ECE-1(-/-); ECE-2(-/-) embryos indicates that proteases distinct from ECE-1 and ECE-2 can carry out ET-1 activation in vivo.     

The JH/LVOH method in the quantification of aortic regurgitation: how the cardiac sonographer may avoid an important potential pitfall

Color flow Doppler allows for methods in which to quantitate the severity of valvular regurgitation. In particular, the regurgitant jet height/left ventricular outflow tract height (JH/LVOH) method of quantitating the severity of aortic regurgitation has been validated and is routinely used in the adult echocardiography laboratory. A potential pitfall exists in the measurement of the LVOH. This article points out this potential source of error and, in addition, proposes steps that may be taken by the cardiac sonographer to avoid this potential measurement pitfall.     

应用彩色多普勒超声探讨主动脉瓣返流与年龄的关系

本文报告经彩色多普勒超声心动图仪检查的１１４６例临床诊断为正常主动脉瓣成人的左室流出道血流结果，共检出有主动脉瓣返流者７１人。各项检查结果经统计学处理后显示了主动脉瓣返流信号的检出率、返流速度、返流量、超声心动图显示的主动脉瓣增厚，回声增强随着年龄的增长，四者呈正相关。认为这种返流信号可能与随着年龄的增长，心血管病发病率增高，主动脉瓣出现退行性变化有关，提出在老年前期就应有效地防治老年病，以来减缓或防止主动脉瓣膜的退行性变。     

STUDIES ON THE MECHANISM OF ADAPTATION OF INFLUENZA VIRUS TO MICE

1. When strains of influenza A virus which have been isolated in chick embryos are introduced into the mouse lung, the virus multiplies readily and achieves initially a titer which is as high as is even obtained, even after repeated passage. The high initial titer of virus may be unaccompanied by any lethal or visible pathogenic effects; but with four or five mouse passages the agent becomes lethal in high titer and causes extensive pulmonary consolidation, though its capacity to multiply in the lung has not increased. In one example the adaptation to mouse lung was accompanied by increasing capacity to agglutinate guinea pig red cells without a corresponding increase in agglutinating power for chicken cells. Influenza B virus, in preliminary tests, did not behave in a similar fashion. 2. The adaptation of influenza A virus to mice is accompanied by changes in antigenic pattern, as detected by cross-tests with the agglutination inhibition method. Two strains, initially similar, with passage, changed in pattern along divergent paths so that they became not only unlike the parent strains but unlike each other. This finding has important implications for the interpretation of the strain difference problem in human influenza.     

超声波终止小鼠早孕的组织学研究

本文报道了一种应用治疗剂量超声抗鼠早孕的方法。结果显示:用12W/cm~2、16W/cm~2、20W/cm~2三组声强辐照小鼠妊娠第7天在体胚胎着床点的组织学急性改变为胎盘出血,电镜下见胎盘和胚胎细胞多数受到严重损伤和细胞质膜失去连续性、细胞核碎裂等;子宫内膜和平滑肌细胞出现线粒体肿胀、内膜细胞的胞质内有的局灶性坏死.受辐照小鼠继续妊娠至第16天时处死发现受辐照的胚胎100%死亡,光镜下见胎盘、胚胎结构解体、塌陷、吸收,细胞核多数溶解,部分呈核淡染、固缩,碎裂或仅存空的子宫腔,子宫内膜增生,平滑肌无明显改变,电镜下见胚胎和胎盘失去正常细胞结构,多为细胞碎片,子宫内膜细胞和平滑肌细胞未见明显异常。结果表明,孕体较母体组织对超声更敏感,更易受损。同时,也说明超声波抗鼠早孕是可行的。     

Atypical ductus venosus blood flow pattern in fetuses with severe tricuspid valve regurgitation.

We observed seven cases of atypical ductus venosus (DV) blood flow velocity waveform pattern with impairment of systolic forward flow resulting in a notch or a significant reduction in peak velocity during the S-wave in systole. All affected fetuses had severe tricuspid valve regurgitation associated with congestive heart failure and/or cardiac malformations. The decrease in venous systolic forward flow modulates the venous pulsatility indices towards more favorable values and should be considered when fetuses with tricuspid regurgitation are followed by Doppler assessment of the DV. Detection of these changes in the DV flow profile should prompt detailed color Doppler echocardiography with special emphasis on right atrioventricular valve regurgitation.