Detection of Apoptosis in Peripheral Blood Cells of 31 Subjects Affected by Down Syndrome Before and After Zinc Therapy

Thirty-one patients affected by Down syndrome (DS) were investigated to study the presence of apoptosis in peripheral blood cells in relation to the plasma levels of zinc. Twelve patients had undergone therapy with ZnSO₄, while the remaining 19 were untreated. The presence of programmed cell death was evaluated by means of electron microscopy, in situ nick translation (NT), and agarose gel electrophoresis of DNA. These approaches evidenced the presence of apoptosis in peripheral blood cells of patients before therapy with ZnSO₄, while after zinc supplementation there was a reduction in the number of apoptotic cells. These results suggest that the process of programmed cell death in peripheral blood cells of patients with Down syndrome is related to the plasma levels of zinc ion.     

Programmed Cell Death of Peripheral Myeloid Precursor Cells in Down Patients: Effect of Zinc Therapy

Hemopoietic stem cell differentiation represents the primary rule of self-renewal, proliferation, and specialization modulated by several mechanisms, including growth factors, cell interactions, and bioavailability of various ions, especially Ca²⁺ and Zn²⁺. Apoptotic death, during normal cell turnover, has been widely studied and is recognized as an important pathway for clonal deletion in the hemopoietic system. Multi-parametric analyses have shown that subjects with Down syndrome show low levels of plasmic zinc associated with the presence of immature myeloid cells in the peripheral blood. This arrangement is repaired by in vivo zinc therapy. This study presents morphological and biochemical analyses to show that ZnSO₄ therapy induces the disappearance of peripheral myeloid precursor cells by a programmed cell death mechanism. The programmed zinc-therapy-induced cell death presumably provides a simple way to regulate the myeloid differentiation selecting appropriate cells.     

Programmed Cell Death of Peripheral Myeloid Precursor Cells in Down Patients: Effect of Zinc Therapy

Hemopoietic stem cell differentiation represents the primary rule of self-renewal, proliferation, and specialization modulated by several mechanisms, including growth factors, cell interactions, and bioavailability of various ions, especially Ca²⁺ and Zn²⁺. Apoptotic death, during normal cell turnover, has been widely studied and is recognized as an important pathway for clonal deletion in the hemopoietic system. Multi-parametric analyses have shown that subjects with Down syndrome show low levels of plasmic zinc associated with the presence of immature myeloid cells in the peripheral blood. This arrangement is repaired by in vivo zinc therapy. This study presents morphological and biochemical analyses to show that ZnSO₄ therapy induces the disappearance of peripheral myeloid precursor cells by a programmed cell death mechanism. The programmed zinc-therapy-induced cell death presumably provides a simple way to regulate the myeloid differentiation selecting appropriate cells.     

实时荧光定量PCR快速分子诊断唐氏综合征

目的　探讨快速诊断唐氏综合征的检测方法 .方法　采用实时荧光定量PCR技术 ,检测唐氏患者 2 3例、正常人 33例 ,定量分析比较ΔCt值 .结果　正常组和唐氏组ΔCt值有显著性差异 (p <0 .0 0 1) ,并建立了相应的参考值范围 .结论　实时荧光定量PCR具有简单、快速、特异性高等优点 ,可用于快速诊断唐氏综合征     

唐氏综合征筛查高风险人群人口特征学分析

目的对十堰地区唐氏综合征筛查高风险人群进行人口特征学分析,以期发现该类人群的共性,加强对其健康管理,提 高检出率,减少唐氏儿的出生遥方法选择2010 年9 月~2014 年12 月于十堰市妇幼保健院就诊的DS 筛查高风险孕妇进行问 卷调查,并对调查结果进行统计学分析遥结果研究共纳入89 例DS 高风险孕妇,逸35 岁人群高危占比显著高于约35 岁人群 （约0.05）曰多孕多产者尧有异常孕产史者高危占比大于孕产次数少尧无不良孕产史者,但差异无统计学意义遥结论本研究发现 高龄与DS 筛查风险值密切相关,多孕多产史尧异常孕产史与DS 筛查高危有相关趋势,但关系尚不明确遥故临床工作中,应加强 对高龄孕妇的管理,做好其产前筛查工作曰对多孕多产与有异常孕产史妇女也应重视产前筛查遥     

Comparative Analysis of Mycobacterial Heat Shock Proteins-Induced Apoptosis of Peripheral Blood Mononuclear Cells in Sarcoidosis and Tuberculosis

Sarcoidosis (SA) is a granulomatous disorder of an unknown etiology. Mycobacterium tuberculosis heat shock proteins (Mtb-hsp), considered as causative agents, play an important role in apoptosis. A role for apoptosis has been proposed in pathogenesis of SA and tuberculosis (TB) granuloma formation but results remain controversial. Differences in Mtb-hsp-induced apoptosis between SA, TB, and healthy subjects found in this study might put some light on the etiology of SA. Early apoptotic peripheral blood mononuclear cells (PBMC) were determined in 22 SA patients, 20 TB patients, and 20 healthy volunteers by flow cytometry (Annexin-V-FITC). Our results revealed that spontaneous apoptosis of monocytes and CD8⁺ T-cells was comparable between tested groups. Apoptosis of unstimulated CD4⁺ T-cells was significantly lower in TB versus controls and insignificantly lower versus SA. Mtb-hsp- and PHA (Phytohemagglutinin)-induced monocytes apoptosis was significantly lower in TB versus controls and SA. Mtb-hsp-induced CD4⁺ T-cell apoptosis was significantly lower in TB versus controls and SA. There were no differences of PHA-induced CD4⁺ T-cell and CD8⁺ T-cell apoptosis between tested groups. Apoptosis of Mtb-hsp-induced CD8⁺ T-cells was significantly lower in TB and SA versus controls. Analysis of PBMC apoptosis before and after stimulation in each tested group revealed that, in contrast to TB, sarcoid monocytes were resistant to Mtb-hsp- and PHA-induced apoptosis and CD4⁺ T-cells were resistant to PHA- but not Mtb-hsp-induced apoptosis. CD8⁺ T-cell apoptosis, before and after Mtb-hsp or PHA stimulation, was significantly increased in all tested groups. It seems likely that dysregulated apoptosis of CD4⁺ T-cells and resistant apoptosis monocytes may be involved in pathogenesis of SA.     

Motor Development and Neuropsychological Patterns in Persons with Down Syndrome

Neuropsychological research has permitted defining specific cognitive profiles among individuals with mental retardation (MR) of different etiology. Namely, the cognitive profile of people with Down syndrome (DS) is often reported to be characterized by a deficit in language abilities that usually exceed impairments in visual–spatial capacities. However, recent studies have demonstrated a more complex neuropsychological profile in this population, with atypical development in the cognitive and in the linguistic domain. This paper is dedicated to reviewing literature regarding motor, linguistic and cognitive abilities in DS. Our aim is to present evidences supporting the hypothesis that individuals with these syndrome exhibit a peculiar motor development and neuropsychological profile with some abilities more preserved and others more impaired. This finding may have theoretical and practical implications. In fact, a better definition of the cognitive pattern in DS may contribute to understand the nature of MR in general and, also, it may suggests individualized rehabilitation treatment protocols.     

Increased Spontaneous Ex Vivo Apoptosis and Subset Alterations in Peripheral Blood T Cells from Patients with Multiple Sclerosis

  In order to characterize the immunophenotype and the lymphocyte apoptosis in multiple sclerosis (MS) patients, the peripheral blood lymphocytes of 46 MS patients and 12 healthy volunteers were studied by flow cytometry. Immunophenotypic alterations included significant increases in T CD4+ lymphocytes and reductions in the percentages of CD3+ and CD8+ T cells. After 24 h of culture spontaneous apoptosis was increased in almost T lymphocyte subsets from MS patients and it was significantly higher in those patients who had suffered more than two relapses in the two previous years. The incidence of spontaneous apoptosis was not dependent on FasL-Fas interactions and correlated with the percentages of cells positive for active caspases but not with percentages of Fas+ cells. The increased susceptibility to apoptosis of peripheral blood T lymphocytes from MS patients is difficult to reconcile with the previously proposed role of a defective lymphocyte apoptosis in the pathophysiology of MS.Alfredo Prieto and David Díaz contributed equally to this work and are joint first authors.The GENIO-II Group is composed by Pablo Barreiro from Hospital La Paz, Madrid; Clara de Andrés and Ma Luisa Martínez from Hospital Gregorio Marañón, Madrid; Bonaventura Casanova from Hospital La Fe, Valencia; Juan Antonio García-Merino and Rosario Blanco from Hospital Puerta de Hierro, Madrid; Jesús Martín from Hospital Miguel Servet, Zaragoza; Francisco Coret from Hospital Clínico, Valencia; José Carlos Alvarez-Cermeño and Dr José Francisco Plaza from Hospital Ramón y Cajal, Madrid; Eugenia Vilar, Blanca Felgueroso and Julián Benito from Hospital de Móstoles; Asunción Morales Otal and M^a Cruz Gutiérrez del Olmo from Hospital 12 de Octubre, Madrid.     

氢化可的松对人外周血来源树突状细胞的诱导凋亡作用研究

旨在探讨氢化可的松对人外周血单核细胞来源树突状细胞 (DC )的凋亡诱导作用及其表型、功能的变化 ;采用细胞因子体外诱导人外周血来源DC ,加入不同浓度的氢化可的松进行培养 ;以细胞计数、PI染色测定细胞周期、间接荧光表型分析、ELISA法测定DC分泌的IL 12和3H TdR掺入测定DC对T细胞的激发作用 ;实验结果表明 ,外周血贴壁的单核细胞在细胞因子的诱导下可以分化为成熟的DC ,不同浓度 (2 μg/ml～ 5 0 μg/ml)的氢化可的松可下调DC表达的CD80和HLA DR分子 (分别下降 41 1%和 10 9% ) ,并使其分泌IL 12的能力明显下降 (由 38 1pg下降为 0 ) ,DC对自体T细胞的激发功能明显降低 ,而且一定浓度的氢化可的松还可以使DC发生凋亡 (凋亡率可达 5 6 6 % )。糖皮质激素不仅通过下调DC表达的协同刺激分子及其分泌的IL 12从而影响DC对T细胞的激发作用 ,而且可以直接诱导DC发生凋亡 ,从而下调免疫应答的发生。     

高浓度抗CD3单抗诱导人外周血单个核细胞凋亡及细胞因子对其影响

使用抗CD3单抗诱导新鲜分离的人外周血单个核细胞(PBMC)凋亡,同时观察细胞因子对其影响。18h后电镜发现处理组发生典型凋亡改变,DNA电泳出现典型阶梯状条带。TUNEL法流式细胞仪检测发现处理组凋亡发生率39.6％,显著高于对照组(<0.01)。IFN-7/TNF-a可显著增强抗CD3单抗诱导的凋亡。IL-1～IL-3、TNF-=对凋亡无明显影响。CsA可抑制抗CD3单抗诱导的凋亡,但加入IFN-y则可完全逆转这种作用。结果表明抗CD3单抗可诱导PBMC发生凋亡,IFN-Y、TNF-与抗CD3有协同作用。     

外周血单核细胞SOCS-1表达与MODS患者预后关系的研究

目的了解外周血单核细胞SOCS-1表达与多器官功能障碍综合征（MODS）患者预后的关系及临床意义。方法收集24例MODS患者，并采集其外周静脉血，采用淋巴细胞分离液密度梯度离心法分离外周血单核细胞（PBMCs），分别以RT-PCR法及Western-blot法检测PBMCs中SOCS-1的基因及蛋白表达，分析其与预后及MODS评分的关系。结果MODS组中死亡患者PBMCs中的SOCS-1mRNA表达量（0.4938±0.0273）显著低于存活患者（0.5475±0.0289）（P〈0.05），SOCS-1蛋白表达量（0.7924±0.0284）显著低于存活患者（0.8406±0.0407）（P〈0.05）。MODS患者的PBMCs中的SOCS-1mRNA表达量与MODS评分呈显著的负相关关系（r=-0.723,P〈0.01），SOCS-1蛋白表达量与MODS评分呈显著的负相关关系（r=-0.534，P〈0.01）。结论在MODS中，SOCS-1的表达可能起到保护组织避免损伤的作用,SOCS-1表达的减少可能提示患者的预后不良     

Micronucleated lymphocytes in parents of Down syndrome children

Down syndrome (DS) is the most common disease due to an autosomal aneuploidy in live born children and also the major known genetic cause of mental retardation. The risk of a DS pregnancy increases substantially with increasing maternal age. However, several women aged less than 35 years at conception have a child with DS. The micronucleus (MN) assay can identify chromosome breakage or chromosome malsegregation and is an ideal biomarker to investigate genomic instability. The aim of the present study was to determine the frequency of peripheral lymphocytes with MN in the parents of DS individuals. The subjects were 17 couples, 1 father and 9 mothers, and 24 couples who had at least one healthy child formed the control group. For each individual we evaluated the frequency of binucleated micronucleated lymphocytes (BNMN%) as number of binucleated lymphocytes containing one or more MN per 1000 binucleated cells. The mean age of DS parents and controls was 32.6 and 29.8 years, respectively. The frequency of MN in DS parents was significantly higher compared to controls. The higher frequency of MN in DS parents suggests a higher predisposition of DS parents to aneuploidy events in this sample.     

Congenital gastrointestinal defects in Down syndrome: a report from the Atlanta and National Down Syndrome Projects

We report Down syndrome (DS)-associated congenital gastrointestinal (GI) defects identified during a 15 year, population-based study of the etiology and phenotypic consequences of trisomy 21. Between 1989 and 2004, six sites collected DNA, clinical and epidemiological information on live-born infants with standard trisomy 21 and their parents. We used chi-squared test and logistic regression to explore relationships between congenital GI defects and infant sex, race, maternal age, origin of the extra chromosome 21, and presence of a congenital heart defect. Congenital GI defects were present in 6.7% of 1892 eligible infants in this large, ethnically diverse, population-based study of DS. Defects included esophageal atresia/tracheoesophageal fistula (0.4%), pyloric stenosis (0.3%), duodenal stenosis/atresia (3.9%), Hirschsprung disease (0.8%), and anal stenosis/atresia (1.0%). We found no statistically significant associations between these defects and the factors examined. Although not significant, esophageal atresia was observed more often in infants of younger mothers and Hispanics, Hirschsprung disease was more frequent in males and in infants of younger mothers and blacks, and anal stenosis/atresia was found more often among females and Asians.     

Investigation of CBS,MTR, RFC-1 and TC Polymorphisms as Maternal Risk Factors for Down Syndrome

Recent evidence shows that almost 92% of the DS children are born from young mothers, suggesting that other risk factors than advanced maternal age must be involved. In this context, some studies demonstrated a possible link between DS and maternal polymorphisms in genes involved in folate metabolism. These polymorphisms, as well as low intake of folate could generate genomic instability, DNA hypomethylation and abnormal segregation, leading to trisomy 21. We compared the frequency of CBS 844ins68, MTR 2756A>G, RFC-1 80G&gt A and TC 776C>G polymorphisms among 114 case mothers and 110 matched controls, in order to observe whether these variants act as risk factors for DS. The genotype distributions revealed that there were not significant differences between both samples. However, when we proceed the multiplicative interaction analyses between the four polymorphisms described above together with the previously studied MTHFR 677C>T, MTHFR 1298A>C and MTRR 66A>G polymorphisms, our results show that the combined genotype TC 776CC / MTHFR 677TT and TC 776CC / MTR 2756AG were significantly higher in the control sample. Nevertheless, there was no significant association after Bonferroni correction. Our results suggest that maternal folate-related polymorphisms studied here have no influence on trisomy 21 susceptibility in subjects of Brazilian population.     

HMME介导的光动力对兔血管平滑肌细胞生长抑制及诱导细胞凋亡的作用

目的：观察原代培养的兔血管平滑肌细胞在HMME-PDT作用下生长的变化及其死亡方式。方法：常规台盼兰染色观察HMME-PDT作用后6h和24h对细胞的杀伤作用，HMME的浓度为10μg／ml，激光剂帚为2．4～9．6J／cm^2；HE染色法观察PDT后24h细胞形态的变化；流式细胞仪Annexin V／PI双染法检测细胞死亡方式；共聚焦品微镜观察HMME在线粒体的定位。结果：台盼兰法检测结果娃示随着PDT能量密度的增加细胞的存活率逐渐下降，光镜下可见大部分细胞呈死亡或凋亡样改变，Annexin V／PI法检测显示PDT 24h后凋亡率可达50．5％．共聚焦显微镜观察到HMME在线粒体内有分布。结论：HMME-PDT能显著抑制兔血管平滑肌细胞的生长．并使其发生明显的凋亡。     

Detection of Single Antibody-Secreting Cells Generated After in Vitro Antigen-Induced Stimulation of Human Peripheral Blood Lymphocytes

Culture and assay procedures are described for the generation and the subsequent detection of single antigen-specific antibody-secreting cells after in vitro stimulation of human peripheral blood lymphocytes with tetanus toxoid. Simple, specific and sensitive, this new assay-culture system is well suited for the analysis of specific antibody production in man.     

Functional Analysis of Genes Implicated in Down Syndrome: 1. Cognitive Abilities in Mice Transpolygenic for Down Syndrome Chromosomal Region-1 (DCR-1)

Down syndrome occurs every 1/1000 births and is the most frequent genetic cause of mental retardation. The genetic substrate of Down syndrome, an extra chromosome 21, was discovered by Lejeune, half-a-century ago, and the chromosome has been fully sequenced, although the gene(s) implicated in the mental retardation observed with the syndrome are still unknown. Observations of patients with partial trisomy of the 21q22.2 fragment suggest that most of the signs of the syndrome, including mental retardation, could be influenced by the region referred to as the Down Minimal Chromosomal Region-1 (DCR-1) for that reason. Using the extensive syntenies between human chromosome 21 and murine chromosome 16, Smith et al. (1995, 1997) developed transpolygenic mice with human chromosome 21 fragments covering the DCR-1. Here, we explored cognitive performances in mice over-expressing the genes carried by these fragments with the Morris water-maze and fear-conditioning procedures. The 152F7 transpolygenic mice had lower performance levels, compared to non-transgenic and other transgenic mice on most measurements in the water-maze. In fear-conditioning, all transgenic mice recorded lower performance levels compared to controls in the altered context stage. The 230E8, 141G6 and 285E6 mice failed to learn or react when the sound used as the conditional stimulus was added. These results showed that the 152F7 region played a crucial role in cognitive impairment, supporting the hypothesis of DYRK-1A gene involvement. However, the data presented here also suggest that other chromosomal regions within the DCR-1 may be involved in specific cognitive functions.     

极谱氧传感仪测定SOD快速诊断先天愚型及其产前诊断的研究

应用极谱氧传感检测仪，测定红细胞中的ＳＯＤ活性来鉴定Ｄｏｗｎ＇ｓ，探索替代传统的染色体诊断方法。本文检测６２例健康人红细胞的ＳＯＤ为６９３５±４８７ｕ／ｍｌ，３２例Ｄｏｗｎ＇ｓ红细胞的ＳＯＤ为１０６０２±８２９ｕ／ｍｌ。而检测４８例健康新生儿红细胞的ＳＯＤ为５６２７±４６３ｕ／ｍｌ，３６例健康胎儿红细胞的ＳＯＤ为５７０７±４７５ｕ／ｍｌ，５例Ｄｏｗｎ＇ｓ新生儿红细胞的ＳＯＤ为８３０７±２２９ｕ／ｍｌ。研究结果显示Ｄｏｗｎ＇ｓ比健康人的红细胞ＳＯＤ高１．５倍，两者具有显著性差异。本方法检测快速，只需５０分钟，灵敏度高，操作简便，具有实用推广价值。