Cx43、β-catenin、Smo在第二生心区的表达规律及意义

背景：第二生心区对于胚胎心脏发育具有重要意义,其发育异常会导致多种心脏畸形,Cx43基因敲除后第二生心区细胞的形成和增殖减少,但具体原因尚未明确。目的：(1)明确β-catenin、Smo以及Cx43在内胚层与第二生心区的表达模式,观察其有无共表达;(2)探究Cx43与Wnt/β-catenin通路或Shh通路之间是否存在相互作用,共同参与第二生心区的发育。方法：选取胚龄10-12 d的ICR小鼠胚胎石蜡包埋连续切片,进行免疫组织化学染色、苏木精-伊红染色、免疫荧光染色;剥离胚龄11 d小鼠胚胎的原始消化管用于蛋白印迹实验和免疫共沉淀。结果与结论：(1)胚龄10-12 d,Cx43与Isl1在前肠腹侧和心包腔背侧壁的部分间充质细胞呈共表达,Isl1阳性细胞增多的同时Cx43阳性细胞也增多;(2)胚龄10-12 d,Cx43与β-catenin在内胚层腹侧共表达;(3)胚龄10-12 d,Cx43与Smo在内胚层上共表达;(4)免疫共沉淀结果表明Cx43与β-catenin之间存在相互作用,共同参与第二生心区的发育。     

ELISA检测心磷脂抗体在胎儿发育方面的应用

目的:探讨胎儿生长迟缓(IUGR)与抗心磷脂抗体(ACA)的相关性。方法:用酶免疫吸附法ELISA检测5330例正常孕妇血清的ACA,同时对IUGR合并ACA阳性产妇产后胎盘进行免疫荧光检查,了解其结构改变。结果:正常孕妇ACA阳性发生率为2.70%,其中IUGR发生率15.28%,ACA阴性的孕妇IUGR发生率为1.77%,两者比较,差异有极显著性(P<0.01)。新生儿IUGRIgG阳性5例,胎盘免疫荧光检查发现胎盘组织均出现免疫球蛋白荧光抗体和补体阳性着色。结论:ACA阳性是IUGR发生的病因之一,ACA检测给IUGR的病因诊治提供了新的研究途径。     

抗心磷脂抗体与胎儿生长受限的相关性研究

目的 探讨抗心磷脂抗体（aCL）与胎儿生长受限（FGR）发生的关系，为胎儿生长受限的早期诊断和预防提供新的思路。方法对2004年6月～2005年2月广饶县妇幼保健院产科门诊、住院的胎儿生长受限患者20例，对照组40例（非孕期健康育龄妇女加例，孕期健康妇女20例）进行研究，应用ELISA法检测aCL-IgG、aCL-IgM抗体。结果正常非孕组aCL阴性率为0，正常妊娠组为5．0％（1／20），FGR组为35．0％（7／20），显着高于对照组（P=0．043），盖别有统计学意义。结论FGR孕妇血清aCL水平明显升高，提示FGR的发生与aCL阳性有关。     

胎儿超声四腔心切面法联合腹部横切面法诊断胎儿右位心的临床价值

目的探讨四腔心切面法联合腹部横切面法诊断胎儿心脏右位心的临床价值。方法回顾2004-2013年9年244 619例孕妇中超声发现的24例胎儿心脏右位心。结果 24例胎儿心脏右位心,单纯性右位心15例、伴心脏多发畸形5例,伴室间隔缺损2例,伴心外畸形2例。其中单纯性右位心15例中,2例首次产前超声没有发现,在例行产前超声复查中纠正并发现单纯性右位心。结论四腔心切面法联合腹部横切面法是诊断胎儿心脏右位心的最佳手段,而胎儿体位、孕周大小、羊水多少、操作经验都将影响超声判断胎儿心脏右位心的准确性,因此有必要对胎儿进行定期复查,以减少或预防胎儿心脏右位心的漏误诊。     

湛江地区4～8月龄胎儿心、肺微量元素的测定

本文采用原子吸收光谱法,测定了湛江地区4—8月龄水囊引产胎儿尸体心脏和肺脏83例中的Zn、Cu、Fe、Ca、Mg五种微量元素。结果显示,各元素的总体含量心脏依次为Mg>Fe>Ca>Zn>Cu,肺为Mg>Ca>Fe>Zn>Cu。五种元素中镁的含量最高,提示本地区胎儿心肺微量元素的含量与沿海环境有密切关系。     

胎儿心、肝和肾长度预测胎儿发育的可行性探讨

目的探讨南方地区胎儿期体质发育数据,为影像诊断、孕期保健等提供参考数据。方法对741例引产胎儿的肝脏、心脏、肾脏的长度及体重进行测量性分析。结果①胎儿的肝脏、心脏、肾脏的长度及体重增长速率与胎龄增加呈正相关。肝脏长度在24周前、33～37周间变化明显,心脏长度在25周后加速均匀增长,肾脏长度在28周前、33～37周间变化明显与体重增长速度比较一致,29～32周为体重变化敏感期。②胎儿体重与肝心肾脏长度间以及孕周与肝心肾脏长度存在线性关系,其中体重分别与心、肝脏长度间的线性关系较强,肝、心、肾脏长度对胎儿体重和孕周的预测效果较好。结论可考虑利用肝、心、肾脏长度对胎儿体重(间接反映发育状况)进行预测。     

抗心磷脂抗体与胎儿宫内发育迟缓及妊高征的关系

目的　探讨抗心磷脂抗体 (AcA)与病理性妊娠的关系 .方法　胎儿宫内发育迟缓 (IUGR) 144例为观察组A ,妊高征 (PIH) 98例为观察组B ,30例正常孕妇为对照组 .用酶联免疫吸附法 (ELISA)检测血清中的AcA .结果　观察组A的AcA阳性 32例 (2 2 .2 % ) ;观察组B阳性 12例 (12 .2 % ) ;对照组阳性 1例 (3 .3 % ) .观察组A、B分别与对照组抗体阳性率比较有明显差异 .结论　IUGR及PIH发病与体内AcA的存在有关 .     

神经嵴细胞和胰岛因子1阳性细胞与小鼠胚胎心流出道发育的关系

目的 探讨迁移中的细胞视黄酸结合蛋白1(CRABP1)阳性神经嵴细胞、胰岛因子1(ISL-1)、阳性心肌前体细胞与小鼠胚胎心流出道发育的关系.方法 36只胚龄8.5～13d小鼠胚胎心连续石蜡切片,选用抗α-平滑肌肌动蛋白(α-SMA)、抗心肌肌球蛋白重链(MHC)、抗转录因子ISL-1、抗CRABP1和抗磷酸化组蛋白H3(PHH3)抗体,进行免疫组织化学及免疫荧光染色.结果 胚龄8.5～10d,ISL-1阳性心肌前体细胞相继出现在心背系膜、原始咽两侧、头面部、鳃弓核心间充质和心包腔背侧壁间充质,构成心管流出道发育的第二生心区.胚龄11～13d,ISL-1阳性细胞在咽前方聚集,形成特征性锥体形结构,并向升主动脉、肺动脉干及主肺动脉隔延伸.胚龄9d前,神经嵴细胞散在分布于ISL-1阳性细胞之间,流出道远侧端可见少量CRABP1和ISL-1双阳性细胞.胚龄10d,CRABP1阳性神经嵴细胞分布在ISL-1阳性鳃弓核心间充质周围.随着发育,弓动脉等处的神经嵴细胞逐渐失去CRABP1表达,开始表达α-SMA.结论 ISL-1阳性第二生心区是动态结构,胚龄8.5～10d时,在神经嵴细胞配合下,向心管动脉端添加心肌细胞;胚龄11d后,开始向平滑肌方向分化,参与升主动脉、肺动脉干和主肺动脉隔的发育.     

胎儿超声心动图检查在结构筛查发现心外异常时的作用

目的了解在超声胎儿结构筛查发现胎儿心外异常时行胎儿超声心动图检查的必要性。方法回顾性研究2011年1月至12月期间在鼓楼医院产前诊断中心超声室接受胎儿超声心动图检查的367例孕妇,其中355例曾在本院进行过超声胎儿结构筛查,回顾其超声结构筛查发现的阳性结果,评价超声结构筛查发现仅有心外异常的病例有无必要进行胎儿超声心动图检查。结果367例接受胎儿超声心动图检查的病例中,355例曾在本院先接受了超声胎儿结构筛查,247例因发现胎儿异常而进行胎儿超声心动图检查,其中发现心脏合并心外结构异常96例,仅怀疑心脏结构或血流异常102例,仅发现心外结构异常49例。它们进一步超声心动图检查阳性结果分别为85例、85例和31例,阳性率分别为88．5％、83．3％和63．2％。结论超声胎儿结构筛查发现阳性结果,不论是心脏结构异常或心外结构异常,均有必要进行胎儿超声＇心动图检查。     

胎儿骨髓源性亚全能干细胞分离培养及诱导肝细胞分化研究

目的研究分离胎儿骨髓源性亚全能干细胞及体内、外向肝细胞分化的潜能。方法利用密度梯度离心结合免疫磁珠方法分离胎儿骨髓源性亚全能干细胞，体外培养鉴定，诱导分化。制备肝功能衰竭重度联合免疫缺陷小鼠（severe combined immunodeficiency，SCID）模型。肝原位输注106左右CD105^＋细胞，对照组分别输注106左右的CD105^-细胞或同等体积的培养液。移植细胞后2d、7d、1个月、3个月时分别取3只鼠的血清及组织标本行肝功能和病理学、免疫组织化学检测。结果免疫磁珠筛选后的免疫细胞化学检测CD105呈弱阳性表达；细胞在对数生长期的倍增时间为30h左右；约传10代后进入衰退期；SCID鼠移植细胞3个月后用鼠抗人白蛋白抗体检测小鼠肝脏中的人白蛋白，可见有点状或小灶状表达。结论来源于胎儿骨髓的亚全能干细胞可以在体外及肝脏微环境下转化为肝细胞样细胞。因此进一步深入研究组织微环境在细胞转化中的机制有十分重要的意义，将为开发诱导胎儿骨髓亚全能干细胞的多向分化潜能提供理论依据。     

先天性心脏病胎儿的染色体检测分析

目的探讨胎儿先心病的发病与染色体异常的关系。方法我院2009年1月～2011年12月孕中晚期胎儿超声筛查检出心脏异常者共85例,采集羊水或脐血样本,行G显带染色体核型分析。结果 1.非青紫型心脏异常中,室间隔缺损发病率最高,其次为主动脉狭窄、肺动脉狭窄。青紫型心脏异常中,心内膜垫缺损发病率最高,其次是右室双出口和大动脉共干。2.先心病胎儿中32例（37.6%）染色体异常,最常见的为18-三体,其次为21-三体。3.合并心外畸形者44例,其中23例（52.3%）存在染色体异常;不合并心脏外异常者41例,其中9例（22.0%）存在染色体异常,二者比较有显著性差异。结论先天性心脏病的病因中染色体畸变占有相当重要的地位,先心病胎儿的染色体检测结果应作为判断是否保留胎儿以及是否手术矫正的指征。     

一例全前脑畸形胎儿的临床及遗传学分析

目的明确一全前脑畸形胎儿的致病原因,为该家系的遗传咨询提供依据。方法收集胎儿的孕期超声资料,应用全外显子测序技术(whole exon sequencing,WES)检测患儿的致病原因,低深度高通量测序技术对胎儿及其父母进行染色体拷贝数变异(copy number variant,CNV)检测,染色体细胞培养方法分析夫妻双方核型。结果胎儿孕期超声提示胎儿脑部结构异常,经诊断后明确为全前脑畸形。WES结果提示胎儿13号染色体存在约33 Mb片段缺失,缺失区域包含1个单倍剂量敏感基因ZIC2。染色体CNV检测结果提示胎儿13号染色体13q31.1-34区域存在32.32 Mb缺失,而夫妻双方均未发现相同的染色体片段缺失。夫妻双方核型分析结果未发现染色体大的结构改变。结论根据临床资料胎儿确诊为全前脑畸形,遗传学检测明确其致病原因为包含单倍剂量敏感基因ZIC2的13号染色体片段缺失。     

Campylobacter fetus bloodstream infection: risk factors and clinical features

In this paper, we report 21 cases of Campylobacter fetus bloodstream infection observed in our institution over a 9-year period. The median age of the patients was 78 years. Most of them (62%) had a significant underlying disease, such as diabetes, immunodeficiency or cardiovascular disease. The main clinical features were fever with (62% of cases) or without (38%) extra-intestinal symptoms. These included mycotic aneurysm of the abdominal aorta (24%) and cellulitis (19%). Antibiotic treatment was mainly based on amoxicilline-clavulanate (57%) or imipenem (21%), for a median duration of 28 days. A favourable outcome was observed in 72% of cases. Death directly attributable to infection was observed for three patients, due to the rupture of an infected aneurysm or relapsing bloodstream infection with septic shock. All patients initially treated with imipenem had a favourable outcome. This report adds evidence that C. fetus bloodstream infection should be suspected in elderly patients with fever, immunodeficiency and cardiovascular damages. Imipenem seems to be the most active drug, especially in severe cases.     

Neuropathological features in a female fetus with OPHN1 deletion and cerebellar hypoplasia

We report the case of a 33-year-old pregnant woman. The third-trimester ultrasound scan during pregnancy revealed fetal bilateral ventricular dilatation, macrosomia and a transverse diameter of the cerebellum at the 30th centile. A brain MRI scan at 31 weeks of gestation led to a diagnosis of hypoplasia of the cerebellar vermis without hemisphere abnormalities and a non compressive expansion of the cisterna magna. The fetal karyotype was 46,XX. The pregnancy was terminated and array-CGH analysis of the fetus identified a 238 kb de novo deletion on chromosome Xp12, encompassing part of OPHN1 gene. Further studies revealed a completely skewed pattern of X inactivation. OPHN1 is involved in X-linked mental retardation (XLMR) with cerebellar hypoplasia and encodes a Rho-GTPase-activating protein called oligophrenin-1, which is produced throughout the developing mouse brain and in the hippocampus and Purkinje cells of the cerebellum in adult mice. Neuropathological examination of the female fetus revealed cerebellar hypoplasia and the heterotopia of Purkinje cells at multiple sites in the white matter of the cerebellum. This condition mostly affects male fetuses in humans. We report here the first case of a de novo partial deletion of OPHN1, with radiological and neuropathological examination, in a female fetus.     

Unusual fan shaped ossification in a female fetus with radiological features of boomerang dysplasia

We report on a female fetus of 24 weeks whose clinical and radiological findings were compatible with boomerang dysplasia (BD). However, histopathology was unusual with a lateral fan shaped diaphyseal ossification. This has never been described either in typical atelosteogenesis I (AT-I) or in BD. The purpose of this report is to find out if this condition is a separate lethal bone dysplasia or another histological feature of the nosological group of AT-I and BD.     

Distribution of the neural crest cells in the heart of birds: a three dimensional analysis

Summary The distribution of neural crest derived cells (NC) in the heart of quail-chick chimeric embryos was analyzed three-dimensionally after computer reconstruction. During the division of the truncus arteriosus into the aorta and the pulmonary trunk, ventral and dorsal columns of NC-derived cells were found in the truncal swellings. These columns were elongations from the aorticopulmonary (AP) septum. The dorsal column extended more proximally than did the ventral column. Around hatching, NC-derived cells located between the proximal aorta and the pulmonary trunk, differentiated into cartilage and connective tissue. They formed a part of the cardiac skeleton. A small number of NC-derived cells were scattered in the cusps of the arterial valves. Cells derived from the right NC were located around the aorta and the right arch arteries but not around the distal pulmonary trunk and the left arch arteries. At the proximal level, cells derived from the rigth NC were located in both the dorsal and ventral columns. These results suggest that the AP septum is mainly formed by NC-derived cells, right and left NC cells migrating into assigned areas in the heart. Location of two columns of NC-derived cells may support a translocation hypothesis for the AP septum during truncal division.     

Effect of neural crest ablation on development of the heart and arch arteries in the chick

Mesenchymal derivatives of the neural crest contribute to the connective tissues and blood vessels of the pharyngeal arches, and participate in the septation of the outflow tract of the heart. The present study was designed to determine the nature and timing of alterations in the development of the heart and arch arteries subsequent to diminished neural crest contributions. The neural crest contributing to the three caudalmost pharyngeal arches was ablated bilaterally in chick embryos and compared with sham or unoper-ated controls. Heart development was studied by scanning electron microscopy. Arch artery development was studied microscopically after intravascular injection of India ink and clearing of the specimen. Neural crest ablation caused morphological changes in most hearts. Hearts in experimental animals commonly were elongate and were subject to inappropriate development of ventricular and atrial areas. A surgical effect delayed the disappearance of arch arteries one and two, and removal of neural crest produced an additional delay. Neural crest ablation caused failure of arch arteries three, four (right), and six to develop to the proper size in some animals. Survival of those whose sixth arch arteries achieved the proper size caused group measurements to reach normal values again by stage 32. Closure of arch arteries in some animals and maintenance in others produced greater variability in experimental animals than in controls. It is significant that heart morphology was altered before septation of the outflow tract normally occurs. This indicates at the least that another factor, such as altered blood flow, contributes to the abnormal development. Altered flow may result from changes in pharyngeal arch mesenchyme and arch artery endothelium.     

Loss of differentiation features in trypsin separated heart muscle cells

Summary Loss of maturation features is demonstrated for 8-day-old chick embryo heart myocytes, once they have been completely dissociated by trypsin. In support of this statement a total of 65 sections of six isolated cells, fixed while still spherical or during early flattening, were examined under the electron microscope. Trypsin-separated heart muscle cells, even though originating from already differentiated embryonic heart tissue, can therefore in principle be used for differentiation experiments in culture. However, the same cell suspensions also yielded an appreciable quantity of nonisolated cells. In such cell complexes, one can find areas showing well-ordered fibrils and intercalated disks. From 27 sections of a cell pair incidentally transferred into culture undissociated and then fixed while still in the globular state, the fourth and fifth sections, starting from the substrate side of the culture, showed an intercalated disk. Because of its small diameter, this cell complex would hardly have been retainable by a gauze with meshes likely to allow passage of only single cells. Thus the availability of differentiation experiments in culture, starting with already differentiated heart tissue, is restricted to cases where, in a selected territory, each cell has been established without doubt as isolated.Supported by grants from the Deutsche Forschungsgemeinschaft.Dedicated to Professor Dr. O. Bucher, Director of the Institute of Histology and Embryology, on the occasion of his sixty-fifth birthday.     

Cardiac neural crest in zebrafish embryos contributes to myocardial cell lineage and early heart function.

Myocardial dysfunction is evident within hours after ablation of the cardiac neural crest in chick embryos, suggesting a role for neural crest in myocardial maturation that is separate from its role in outflow septation. This role could be conserved in an animal that does not have a divided systemic and pulmonary circulation, such as zebrafish. To test this hypothesis, we used cell marking to identify the axial level of neural crest that migrates to the heart in zebrafish embryos. Unlike the chick and mouse, the zebrafish cardiac neural crest does not originate from the axial level of the somites. The region of neural crest cranial to somite 1 was found to contribute cells to the heart. Cells from the cardiac neural crest migrated to the myocardial wall of the heart tube, where some of them expressed a myocardial phenotype. Laser ablation of the cardiac premigratory neural crest at the three- to four-somite stage resulted in loss of the neural crest cells migrating to the heart as shown by the absence of AP2- and HNK1-expressing cells and failure of the heart tube to undergo looping. Myocardial function was assessed 24 hr after the cardiac neural crest ablation in a subpopulation of embryos with normal heart rate. Decreased stroke volume, ejection fraction, and cardiac output were observed, indicating a more severe functional deficit in cardiac neural crest-ablated zebrafish embryos compared with neural crest-ablated chick embryos. These results suggest a new role for cardiac neural crest cells in vertebrate cardiac development and are the first report of a myocardial cell lineage for neural crest derivatives.