EZH2对先天性巨结肠结肠组织中GFRα1表达的影响

目的 通过检测先天性巨结肠（HSCR）患儿结肠组织内胶质细胞源性神经营养因子受体α1（GFRα1）及果蝇Zeste基因增强子人类同源物2（EZH2）的表达水平,探讨EZH2在调控GFRα1基因表达及HSCR发病过程中的作用。方法 随机选取24例行巨结肠根治术的HSCR患儿,取痉挛段结肠组织为试验组。以同期18例因新生儿坏死性小肠结肠炎而接受手术治疗的患儿,取手术切除的坏死结肠组织作为对照组。利用实时荧光定量PCR及Western blot检测两组结肠组织内GFRα1、EZH2的表达水平。将人神经母细胞瘤细胞SHSY5Y分为EZH2过表达组和阴性对照组,EZH2过表达组转染pCMV6-EZH2质粒,阴性对照组转染pCMV6质粒,检测转染后细胞中GFRα1、EZH2的表达水平。结果 与对照组相比,试验组GFRα1及EZH2 mRNA和蛋白的表达水平降低（P < 0.05）,且EZH2蛋白表达水平与GFRα1蛋白呈正相关（r=0.606,P=0.002）。与阴性对照组相比,EZH2过表达组EZH2及GFRα1表达水平明显上调（P < 0.05）。结论 HSCR患儿结肠组织中EZH2低表达可能是GFRα1表达不足,诱发HSCR的原因之一。     

肠神经系统发育的研究进展北大核心CSCD

肠神经系统是由肠神经元和神经胶质细胞组成的网络体,它起源于神经嵴干细胞。而神经嵴干细胞在多种细胞因子及信号分子的参与下,经历增殖、迁移及分化过程,形成各种类型的肠道细胞。这些细胞分别表达不同的神经递质和神经肽,共同调控肠道的功能。胚胎期肠神经系统的发育障碍是先天性巨结肠症(Hirschsprung’s disease,HSCR)的根本原因。近年来,胚胎期肠管神经发育的研究取得了一些进展,尤其是在信号通路及其与HSCR的关系上,这也为HSCR的治疗提供了一些借鉴。     

儿童功能性构音障碍的分子遗传学研究

遗传因素是造成功能性构音障碍的重要原因。本文综述了最近发现的与功能性构音障碍相关的一些基因和染色体区域。详细介绍了FOXP2基因的结构、表达和功能,以及FOXP2基因在发育性言语失用中的变异。作为重要的转录调控因子,FOXP2基因可以调控很多基因的表达。其中CNTNAP2基因是FOXP2基因的重要靶基因,是影响语言及言语发育的重要基因。功能性构音障碍可能发展为阅读障碍,阅读障碍的候选基因也成为研究功能性构音障碍的重要候选基因。一些与阅读障碍相关的染色体区域3p12-13、15q11-21、6p22 及1p34-36被认为可能与功能性构音障碍相关。位于这部分染色体区域的一些基因如ROBO1基因、ZNF280D 基因、TCF12 基因、EKN1基因、KIAA0319 基因等成为研究功能性构音障碍的候选基因。     

孤独症谱系障碍易感基因相关研究进展

孤独症谱系障碍(ASD)是神经发育过程中的一种发育障碍性疾病,是多个易感基因参与发病的多基因疾病。目前已报道的易感基因有100 多个,相关研究包括易感基因的染色体位点研究、易感基因筛查研究和易感基因的表观遗传学异常。已报道的易感基因编码的蛋白质有:神经细胞粘着分子;离子通道蛋白;支架蛋白;蛋白激酶、受体、载体;信号通路调控蛋白以及昼夜节律相关蛋白。易感基因突变和表达调控机制的研究进展有助了解ASD 的遗传参与的发病机制,可望能给ASD 的诊断和治疗提供新的思路。该文对ASD 易感基因方面的研究现状进行了综述。     

钙库操纵性钙通道在先天性巨结肠结肠平滑肌层表达改变的初步研究

目的观察在先天性巨结肠(Hirschsprung's disease,HSCR)结肠平滑肌上钙库操纵性钙通道(store-operated calcium channel,SOCC)的相关的基因及蛋白Orai1和Stim1表达,初步探索SOCC与HSCR结肠动力障碍机制的相关性。方法选择2017年4月至2018年7月在上海交通大学医学院附属新华医院及嘉兴市妇幼保健院接受手术治疗的HSCR患儿24例。其中,男17例,女7例,均为常见型巨结肠;年龄为(8.6±3.6)个月,范围为6~14个月。采用蛋白质印迹法、荧光定量PCR、免疫组织化学等技术,在24例HSCR患儿手术切除的近端切缘段(正常段)、狭窄段和扩张段结肠平滑肌标本中检测与SOCC相关的基因及蛋白Orai1和Stim1的表达情况。结果蛋白质印迹法结果显示24例HSCR患儿结肠狭窄段Orai1及Stim1蛋白的表达水平与扩张段相比明显增加,差异具有统计学意义(t=2.746,P=0.012;t=2.779,P=0.011);扩张段Orai1及Stim1蛋白的表达水平与近端切缘段相比也明显增加,差异具有统计学意义(t=2.296,P=0.031;t=2.403,P=0.025);狭窄段Orai1及Stim1蛋白的表达水平显著高于近端切缘,差异具有统计学意义(t=5.915,P<0.01;t=7.726,P<0.01)。荧光定量PCR结果显示和扩张段相比,狭窄段Orai1和Stim1的mRNA的表达明显增加,差异具有统计学意义(t=2.330,P=0.029;t=2.698,P=0.013);扩张段Orai1和Stim1的mRNA的表达也较近端切缘明显增加,差异具有统计学意义(t=2.425,P=0.024;t=2.186,P=0.039);狭窄段Orai1及Stim1的mRNA的表达显著高于近端切缘,差异具有统计学意义(t=7.332,P<0.01;t=18.052,P<0.01)。免疫组织化学结果也显示与扩张段相比,Orai1和Stim1蛋白在狭窄段的表达明显增加,扩张段Orai1和Stim1蛋白的表达较近端切缘段亦明显增加。结论HSCR患儿结肠病变部位平滑肌层的SOCC的表达明显增强,说明该信号通路的异常可能与HSCR的结肠动力障碍机制相关。     

钙库操纵性钙通道在先天性巨结肠结肠平滑肌层表达改变的初步研究

目的观察在先天性巨结肠(Hirschsprung’s disease,HSCR)结肠平滑肌上钙库操纵性钙通道(store-operated calcium channel,SOCC)的相关的基因及蛋白Orai1和Stim1表达,初步探索SOCC与HSCR结肠动力障碍机制的相关性。方法选择2017年4月至2018年7月在上海交通大学医学院附属新华医院及嘉兴市妇幼保健院接受手术治疗的HSCR患儿24例。其中,男17例,女7例,均为常见型巨结肠;年龄为(8.6±3.6)个月,范围为6～14个月。采用蛋白质印迹法、荧光定量PCR、免疫组织化学等技术,在24例HSCR患儿手术切除的近端切缘段(正常段)、狭窄段和扩张段结肠平滑肌标本中检测与SOCC相关的基因及蛋白Orai1和Stim1的表达情况。结果蛋白质印迹法结果显示24例HSCR患儿结肠狭窄段Orai1及Stim1蛋白的表达水平与扩张段相比明显增加,差异具有统计学意义(t=2.746,P=0.012;t=2.779,P=0.011);扩张段Orai1及Stim1蛋白的表达水平与近端切缘段相比也明显增加,差异具有统计学意义(t=2.296,P=...     

基于生物信息学筛选唐氏综合征胎儿脑病变相关基因及信号通路北大核心CSCD

目的基于生物信息学分析,筛选唐氏综合征(DS)胎儿脑病变相关基因及信号通路,探究其在DS神经病变发生发展中的潜在作用机制。方法回顾性研究。2021年12月从基因表达数据库下载数据集GSE59630,利用R软件筛选DS和正常胎儿脑组织之间的差异表达基因(DEGs),并对其进行基因本体(GO)、京都基因与基因组百科全书(KEGG)富集分析和基因集富集分析(GSEA)。利用基因相互作用检索工具在线数据库和Cytoscape软件构建蛋白互作(PPI)网络,确定核心模块及核心基因。采用实时定量聚合酶链反应在3对22~33周龄DS和正常胎儿脑组织中验证神经退行性变相关核心基因的表达变化。结果从DS和正常胎儿脑组织中共筛选出225个DEGs,其中18个为上调基因,207个为下调基因。GO功能富集分析显示DEGs主要富集在神经发生、神经元凋亡、转录调控、线粒体能量代谢等过程,KEGG通路富集分析显示DEGs与多种神经退行性疾病有关,GSEA提示细胞凋亡、炎症反应在DS神经病变发生中发挥重要作用。根据PPI网络筛选出10个核心基因,主要与组蛋白乙酰化和转录调控相关。经组织验证,其中RAB8A、TBP、TAF6表达变化趋势与芯片数据相一致。结论通过胎儿脑组织转录组学分析筛选出的核心基因及关键信号通路,有助于全面了解DS神经病变发生的分子机制,为探索DS神经发育异常和智力低下的临床诊断及治疗靶点提供了新的方向。     

基于生物信息学分析microRNA-495-5p在早产儿支气管肺发育不良中的表达及其临床意义

目的 探讨支气管肺发育不良（BPD）早产儿血清中microRNA-495-5p（miRNA-495-5p）的表达变化并对其进行生物信息学分析，为深入研究miRNA-495-5p与BPD的关系提供理论依据。方法 收集2015年1月至2016年12月NICU住院治疗早产儿的一般临床资料，选取具有早期BPD临床表现的20例患儿为BPD组，无早期BPD临床表现的20例患儿为对照组。采集两组患儿末梢外周血，每组各随机选取5例患儿，应用miRNA芯片技术筛选两组患儿血清中差异性表达的miRNAs；每组各随机选取6例患儿，采用RT-PCR技术再次验证其差异性表达。应用TargetScan、miRDB、miRWalk数据库对miRNA-495-5p进行靶基因预测；采用DAVID数据库对靶基因进行基因功能富集分析和信号转导通路富集分析。结果 与对照组患儿相比，BPD组患儿血清miRNA-495-5p表达显著上调（P < 0.05）。通过3种数据库预测miRNA-495-5p的靶基因共有117个，其靶基因功能分别富集于转录调节活性、转录激活活性、转录辅助激活活性等分子功能，代谢过程的调控、依赖DNA的转录调控、血管模式等生物学过程，以及核质、膜组分、不溶性组分等细胞组分上（P < 0.05）；信号转导通路则显著富集于mTOR信号通路中（P < 0.05）。结论 miRNA-495-5p可能通过调控新生血管生成、干细胞分化、细胞凋亡及自噬等参与BPD的发生发展，为后续深入研究其在BPD中的作用及功能机制提供了重要线索。     

Neurexin、Neuroligin基因在人类及鼠类动物肠神经系统的表达及意义

目的 探讨Neurexin和Neuroligin基因在人类及鼠类动物肠神经系统(ENS)是否有表达,以及对ENS突触结构与功能的作用,探查其在先天性巨结肠各段的表达特征及意义.方法 首先制备小鼠、大鼠、豚鼠、正常人体肠管肌间神经丛铺片,通过双重免疫荧光细胞染色,证明Ne-urexin、Neuroligin基因是否在肠神经系统表达,再采用免疫组织化学技术对20例先天性巨结肠狭窄段,移行段和扩张段肌间神经丛铺片标本染色,判断Neurexin、Neuroligin基因的表达情况.结果①Neurexin、Neuroligin蛋白在小鼠、大鼠、豚鼠、人体的肠肌间神经丛与神经细胞标记Hu在同一细胞表达;而Neurexin蛋白与突触前标记物synaptophysin,Neuroligin蛋白与突触后标记物PSD95也在同一神经组织出现;②Neurexin、Neuroligin蛋白在先天性巨结肠扩张段表达最多,移行段明显减少(P<0.05),病变段无表达;同一组织Neurexin和Neuroligin的表达无明显差异(P>0.05).结论 Neurexin、Neuroligin基因在鼠齿类动物及人体肠神经系统都有表达,不仅在胞体,而且在轴突、树突都有表达,Neurexin在突触前表达,Neuroligin在突触后表达.它们对肠神经系统突触的结构功能起重要作用,与一些胃、肠道动力性疾病有密切关系,是导致先天性巨结肠蠕动功能丧失的重要原因.     

X连锁多内分泌腺病肠病伴免疫失调综合征一例基因及蛋白表达研究

目的 探讨表现为顽固性腹泻、皮疹以及有或无胰岛素依赖性糖尿病的疑似X连锁多内分泌腺病肠病伴免疫失调综合征(IPEX)患儿FOXP3基因变异及其蛋白表达水平.方法 对近两年来我院收治的4例表现为早发性顽固性腹泻、皮疹以及有或无胰岛素依赖性糖尿病的疑似IPEX男性患儿进行FOXP3基因扩增及测序分析,将发现的可疑突变位点通过数据库查询及与100例健康儿童相同位点序列比较,采用流式细胞仪检测CD4~+CD25~+FOXP3~+调节性T细胞比例和FOXP3蛋白表达.结果 4例疑似患儿中发现1例FOXP3突变,为11号外显子66位碱基错义突变(G>A),导致FOXP3蛋白370位氨基酸由甲硫氨酸替换为异亮氨酸(Met370Ile),患儿母亲为携带者.100例正常儿童FOXP3基因相同位点未见变异,故可排除该位点多态性可能.该突变为此前未见报道的新突变,患儿CD4~+CD25~+FOXP3~+调节性T细胞比例升高而FOXP3表达量无减低.结论 通过临床、免疫学筛查和基因分析,发现我国首例IPEX患儿(g:13128 G>A c:1298 G>A Met370Ile),对早发胰岛素依赖性糖尿病、顽固性腹泻及不明肾脏损害婴幼儿,应考虑IPEX可能并进行FOXP3基因分析.     

Allergic factors associated with the development of asthma and the influence of cetirizine in a double-blind, randomised, placebo-controlled trial: First results of ETA®

There is a common progression known as the allergic march from atopic dermatitis to allergic asthma. Cetirizine has several antiallergic properties that suggest a potential effect on the development of airway inflammation and asthma in infants with atopic dermatitis. Methods. Over a two year period, 817 infants aged one to two years who suffered from atopic dermatitis and with a history of atopic disease in a parent or sibling were included in the ETAC^® (Early Treatment of the Atopic Child) trial, a multi-country, double-blind, randomised, placebo-controlled trial. The infants were treated for 18 months with either cetirizine (0.25mg/ kg b.i.d.) or placebo. The number of infants who developed asthma was compared between the two groups. Clinical and biological assessments including analysis of total and specific IgE antibodies were performed. Results. In the placebo group, the relative risk (RR) for developing asthma was elevated in patients with a raised level of total IgE (≥ 30 kU/I) or specific IgE (≥ 0.35 kUA/I) for grass pollen, house dust mite or cat dander (RR between 1.4 and 1.7). Compared to placebo, cetirizine significantly reduced the incidence of asthma for patients sensitised to grass pollen (RR = 0.5) or to house dust mite (RR = 0.6). However, in the population that included all infants with normal and elevated total or specific IgE (intention-to-treat - ITT), there was no difference between the numbers of infants developing asthma while receiving cetirizine or placebo. The adverse events profile was similar in the two treatment groups. Discussion. Raised total IgE level and raised specific IgE levels to grass pollen, house dust mite or cat dander were predictive of subsequent asthma. Cetirizine halved the number of patients developing asthma in the subgroups sensitised to grass pollen or house dust mite (i.e. 20% of the study population). In view of the proven safety of the drug, we propose this treatment as a primary pharmacological intervention strategy to prevent the development of asthma in specifically sensitised infants with atopic dermatitis.     

Resurgence of measles in Singapore: profile of hospital cases

OBJECTIVE: To ascertain the profile of cases of measles seen at a general hospital during a recent outbreak that occurred despite a measles vaccination program. METHODOLOGY: A retrospective study from January 1991 to March 1998. All patients with measles (ICD code 055. 9) seen at the emergency unit or as inpatients were included. RESULTS: There were 87 cases identified. The diagnosis was clinical in all and proven serologically in 71%. Eighty-five per cent of the cases occurred between January 1997 and March 1998. There was a bi-modal age distribution with peaks in the very young (相似文献   

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孤独症谱系障碍(autistic-spectrum disorders,ASDs)近年来患病率逐年攀升至1%左右,其症状往往伴随终生,成为严重威胁儿童健康和发展的神经发育性疾患;注意缺陷多动障碍(attention deficit hyperactivity disorder,ADHD)是儿童期最常见的精神障碍,国内报道患病率为4.13%～5.83%,其症状可延续至青少年期,甚至到成年期[1]。这两类精神障碍在成年期的临床表现、共患病、治疗策略和预后与儿童期有哪些不同呢?本文通过回顾相     

EXCITING NEW TREATMENT APPROACHES FOR PATHYPHYSIOLOGIC MECHANISMS OF SICKLE CELL DISEASE

During the past several decades, our understanding of the complex pathophysiology of vasoocclusion associated with sickle cell disease has improved greatly. Interaction of genes, hemoglobin molecules, red cell membrane and metabolic changes, cell-cell interactions and cell-plasma interactions, red cell adhesion to vascular endothelium, activation of coagulation, and vascular reactivity play a role in vaso occlusion. Penicillin prophylaxis of pneumococcal infections and appropriate use of blood transfusions and other supportive measures improved survival of sickle cell patients. Hydroxyurea made a major impact on sickle cell therapy when it was shown to decrease acute painful episodes, acute chest syndrome, and the need for blood transfusion in adults. Significant experience in the use of hydroxyurea has been accumulated in older children. The benefits and risks of hydroxyurea for younger children and long-term risks in all patients will be evaluated in future investigations. Other promising therapies include butyrate compounds, clotrimazole, magnesium supplementation, poloxamer 188, antiadhesion agents, anticoagulant approaches, and nitric oxide. Hemopoietic transplantation remains the only curative therapy. However, several transgenic mouse models are available for studies of gene therapy or other treatment approaches on biochemical, cellular, and pathologic effects of mutant genes.     

Infiltrations of Epstein-Barr virus-carrying cells in granular lymphocyte-proliferative disorders corresponding to chronic active Epstein-Barr virus infection

A 21-year-old man with granular lymphocyte-proliferative disorders (GLPD) associated with chronic active Epstein-Barr virus (EBV) infection is described. Chromosomal analyses revealed several clonal abnormalities and two of them were mainly repetitious. High copy numbers of monoclonal EBV genome were also detected in the proliferative large granular lymphocytes (LGLs), indicating the monoclonal expansion of EBV-infected LGLs. The patient had an indolent course for several years, and there was no evidence of infiltrations of his bone marrow until the end stage. At autopsy, microscopic studies revealed marked infiltrations of LGL in the liver and spleen, and the infiltrating cells were NK-cell immunophenotype. The infiltrated LGLs showed latency I.     

LUTEINIZING HORMONE RECEPTOR MUTATIONS IN DISORDERS OF SEXUAL DEVELOPMENT AND CANCER

Human male sexual development is regulated by chorionic gonadotropin (CG) and luteinizing hormone (LH). Aberrant sexual development caused by both activating and inactivating mutations of the human luteinizing hormone receptor (LHR) have been described. All known activating mutations of the LHR are missense mutations caused by single base substitution. The most common activating mutation is the replacement of Asp-578 by Gly due to the substitution of A by G at nucleotide position 1733. All activating mutations are present in exon 11 which encodes the transmembrane domain of the receptor. Constitutive activity of the LHR causes LH releasing hormone-independent precocious puberty in boys and the autosomal dominant disorder familial male-limited precocious puberty (FMPP). Both germline and somatic activating mutations of the LHR have been found in patients with testicular tumors. Activating mutations have no effect on females. The molecular genetics of the inactivating mutations of the LHR are more variable and include single base substitution, partial gene deletion, and insertion. These mutations are not localized and are present in both the extracellular and transmembrane domain of the receptor. Inactivation of the LHR gives rise to the autosomal recessive disorder Leydig cell hypoplasia (LCH) and male hypogonadism or male pseudohermaphroditism. Severity of the clinical phenotype in LCH patients correlates with the amount of residual activity of the mutated receptor. Females are less affected by inactivating mutation of the LHR. Symptoms caused by homozygous inactivating mutation of the LHR include polycystic ovaries and primary amenorrhea.     

A profile of respiratory symptoms in urban and rural South Australian school children

This report describes the cross-sectional analyses of data from the first year of a longitudinal study using questionnaire and respiratory function data over a 5 year period from a sample of rural South Australian school children. The cumulative or lifetime prevalences of respiratory symptoms were estimated in 825 rural and 1261 urban school children aged between 5 and 15 years in order to determine if the prevalence rates differed between rural and urban school children. The study found the overall cumulative prevalence of asthma and/or wheezy breathing (AWB) to be 24.1% in the rural school children compared to 27.6% in the urban school children. Most children developed AWB symptoms before the age of 7 years, with 20% reporting moderately severe symptoms and 10% having more than one attack per fortnight. The cumulative prevalence of bronchitis, loose/rattly cough (BLRC) differed significantly between the rural school children (34.1%) and urban school children (47.9%). The BLRC symptoms preceded the development of AWB in many cases. Urban school children also reported a higher prevalence of atopic conditions.     

Personality profiles and heart rate variability (vagal tone) in children with recurrent abdominal pain

The aim of the study was to explore psychological factors and autonomic activity in children with recurrent abdominal pain and to compare them with those in a control group of healthy children. The Personality Inventory for Children was used for assessment of developmental, emotional and psychosocial factors in 25 children with recurrent abdominal pain (age, 7-15 y). Parasympathetic and sympathetic functions in these children and in 23 healthy control subjects (age, 7-13 y) were also investigated, non-invasively using a computerized polygraph. Vagal tone (parasympathetic function) was indexed by calculation of respiratory sinus arrhythmia in beats/min. Skin conductance (sympathetic function) was recorded by the constant current method. On the Personality Inventory for Children, 16 patients had high scores on somatic concern. Several patients had scores in the clinical range for depression, withdrawal and anxiety, but the mean scores for these personality profile scales were well within the normal range of healthy children. Interestingly, there was a spike on the L (Lie)-scale for most of the patients and 15 patients had scores above or close to the clinical cut-off value. As compared with the scores in healthy children, vagal tone and sympathetic tone were normal. Conclusion: Many children with recurrent abdominal pain have scores in the clinical range for depression, withdrawal, anxiety and L-scale indicating coping problems, denial and a trend towards somatic concern that may contribute to the evolution of abdominal pain. Autonomic nerve activity was not disturbed in these children.     

Hauttemperaturen verschiedener Körperoberflächenareale des Rumpfes bei Säuglingen

Summary In two groups of infants (3–53 weeks old) skin temperatures were controlled in different areas of the trunk—i.e.: regions of sternum, lungs, heart, liver, spleen, kidneys—at different room-temperatures (group I: 21–25°C; group II: 29–32°C). Rectal temperatures of some probands in both groups also had been controlled simultaneously. A definite change in the reaction to heat was proofed in different periods of the first year of life. In higher environmental temperatures the skin temperature was almost constant at every controll-point of the skin, even in older infants. In lower environmental temperatures the skin temperatures lowered continuously with age till 7. to 9. moth. From 10. to 12. month the lowering of skin temperature discontinued. The rectal temperatures were relatively constant in all infants. Only in infants from 7. to 12. month, whose skin temperatures were controlled in lower as well as in higher environmental temperatures, a tendency to higher rectal temperatures was proofed in warmer environmental temperatures.The significance of these results is discussed.

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Untersuchungen mit Unterstützung durch die Deutsche Forschungsgemeinschaft.