高浓度氧对早产鼠肺一氧化氮合酶基因表达的影响

目的　探讨高浓度氧 (简称高氧 )对早产鼠肺一氧化氮合酶 (NOS)分布及基因表达的影响。方法　将 2 1d孕早产鼠随机分为高氧暴露组 (简称高氧组 )和空气对照组 (对照组 ) ,分别置于常压高氧仓中 (氧体积分数 >0 95 )和正常空气中暴露 7d ,采用逆转录 聚合酶链反应 (RT PCR)、免疫蛋白分析和免疫组织化学染色观察NOS分布及基因表达。此外对肺组织干 /湿重比值 ,支气管肺泡灌洗液 (BALF)成份和肺病理组织变化也进行了对比分析。结果　高氧组与对照组比较 ,早产鼠肺组织有明显水肿、出血和炎症 ;高氧组支气管肺泡灌洗液中蛋白含量 (中位数为 1 4 9g/L)、细胞数 (中位数为 139 70× 10 7/L)和肺组织干 /湿重比值 (5 5 7± 0 2 9)较对照组 (分别为 :0 32g/L、16 30× 10 7/L、5 2 9± 0 2 5 )均明显增加 (t=2 8、2 1、2 2 9,P均 <0 0 5 ) ;高氧组肺组织内皮细胞型一氧化氮合酶 (eNOS)mRNA、eNOS和诱导型一氧化氮合酶 (iNOS)蛋白表达 (分别为 :1 0 2± 0 0 6、8 77± 0 75、4 6 1± 0 6 5 )较对照组 (分别为 :0 70± 0 12、4 5 2± 1 0 2、3 2 4± 0 5 5 )明显增加 (t =6 36、8 14、3 2 1,P <0 0 1、P <0 0 1、P <0 0 5 )。iNOSmRNA也显示有增加的趋势 ,但差异无统计学意义 ,同时免疫组织化     

高氧对新生大鼠肺caspase-3和p53基因表达及肺细胞凋亡的影响

目的探讨高氧对新生大鼠肺caspase3和p53基因表达及肺细胞凋亡的影响。方法采用SpraqueDawley新生大鼠95%氧气暴露建立高氧肺损伤模型。应用RTPCR技术检测肺组织caspase3mRNA和p53mRNA水平,凝胶电泳条带用成像系统照相分析结果。计算目的基因PCR产物条带与内参照βactin条带光密度值的比值,作为p53基因的相对表达量,结果以x±s标记,而caspase3mRNA表达量则以阳性表达或阴性表达为标记。应用脱氧核糖核酸转移酶介导的细胞凋亡标记技术(TUNEL)原位检测细胞凋亡。光镜下随机计算5个视野中500个肺细胞中的阳性细胞数,结果以x±s标记。结果新生大鼠暴露于95%氧浓度环境中24h后肺组织中p53mRNA表达中度增加(q=3.2305,P>0.05),48h后表达显著增加,与空气对照组相比差异有统计学意义(q=7.2941,P<0.05)。新生鼠高氧处理72h、96h后,肺组织p53mRNA表达又恢复到正常水平。在各空气对照组和各高氧处理组中个别新生鼠肺的caspase3mRNA有微量表达,绝大多数新生鼠肺的caspase3mRNA没有表达,差异无统计学意义。95%氧暴露7天的新生鼠肺细胞凋亡水平明显高于空气暴露组新生鼠肺细胞凋亡水平,两者比较差异有极显著的统计学意义(F=100,P<0.001)。结论在高浓度供氧下,肺组织通过暂时上调p53基因的表达,介导细胞周期停滞,阻止G0/G1期细胞进入S期,抑制细胞分裂、增殖,同时p53促进细胞凋亡,从而导致肺生长发育受阻和肺损伤。新生鼠暴露于95%氧环境中,肺组织caspase3基因基本上不表达,因此推测高氧肺细胞凋亡可能存在不经过caspase3的凋亡途径。     

泛素-蛋白酶体途径在高体积分数氧肺损伤大鼠早期的激活

目的 探讨高体积分数氧(高氧)暴露早期大鼠肺损伤情况,以及泛素-蛋白酶体途径是否激活.方法 实验采用完全随机设计,SD大鼠20只随机分为高氧组和空气组,每组10只.空气组大鼠持续吸入空气(氧体积分数210 ml/L)72 h;高氧组大鼠持续吸入高氧(氧体积分数950 ml/L)72 h制备高氧肺损伤模型.并视肺形态学变化并确定大体病理分级,肺损伤病理评分;测定肺湿/干质量比值.免疫组织化学方法和Western blot方法检测肺组织泛素化蛋白的表达和蛋白酶体20 s的活性,评价泛素-蛋白酶体途径激活情况.结果 1.成功复制高氧肺损伤大鼠模型.2.大鼠高氧暴露后,出现明显肺水肿,肺湿/干质量比值增高(P=0).3.高氧组大鼠高氧暴露72 h肺组织鲜红色,饱满状,表面有点片状出血或局灶状出血,胸腔有少量积液.高氧组大体病理分级与空气组比较差异有显著性(P=0.005).光镜观察高氧组肺泡上皮肿胀,肺泡壁增宽,毛细血管扩张和充血,肺泡间质和肺泡腔水肿明显,肺泡内炎性细胞增多.高氧组肺损伤病理评分较空气组显著增高(P=0).4.免疫组织化学和Western blot结果均显示大鼠高氧暴露72 h肺组织泛素化蛋白表达显著增强(P=0).5.高氧组蛋白酶体20 s活性较空气组明显升高(P=0).结论 高氧暴露72 h对大鼠肺脏有明显损伤,弥散性肺泡上皮细胞、肺血管内皮细胞损伤,肺内炎性反应细胞浸润伴肺水肿为主要病理学改变.高氧暴露早期大鼠肺组织泛素-蛋白酶体途径激活,并参与肺损伤病理生理过程.     

p38丝裂素活化蛋白激酶在新生大鼠高氧肺损伤中的作用

目的探讨p38丝裂素活化蛋白激酶(p38MAPK)在新生大鼠高氧肺损伤中的表达及其特异性抑制剂SB203580,对其产生保护作用的机制。方法160只新生大鼠随机分为空气对照组、高氧肺损伤组、高氧肺损伤 SB203580组和高氧肺损伤 生理盐水组,建立模型。作用12、24、72h和1周后,分别处死大鼠。取右上肺进行肺组织病理学检查,取右下肺进行肺湿/干重比值(W/D)测定,取左肺以Western免疫印迹法检测肺组织中p38MAPK的表达,以酶联免疫吸附试验检测肺组织中TGF-β1的含量。结果72h时,高氧肺损伤组和高氧肺损伤 生理盐水组p38MAPK呈强阳性表达;在这两组中,肺组织TGF-β1浓度随时间延长呈持续上升趋势,在各时相点均明显高于空气对照组和高氧肺损伤 SB203580组(P均<0.01)。结论p38MAPK参与了新生大鼠高氧肺损伤的过程,SB203580能够通过阻断p38MAPK表达,进而抑制TGF-β1表达来减轻这种损伤。     

地塞米松对高氧肺损伤新生大鼠肺水通道蛋白1表达的影响

目的 探讨地塞米松对新生大鼠高氧肺损伤时水通道蛋白1(AQP1)表达的影响及其对肺损伤的可能保护机制.方法 新生Wistar大鼠32只随机分为空气组、高氧组、空气+地塞米松组、高氧+地塞米松组.第3天取肺组织,采用逆转录-聚合酶链反应(RT-PCR)和免疫组织化学法检测AQP1的mRNA表达和分布变化;并对肺湿/干重比(W/D)、支气管肺泡灌洗液(BALF)中的蛋白含量、肺通透指数及组织病理学改变进行对比分析.结果 高氧暴露第3天肺组织出现出血、炎性细胞浸润和水肿,肺W/D、BALF蛋白含量、肺通透指数明显升高;地塞米松干预组肺损伤程度减轻,测定值降低.空气组、高氧组、高氧+地塞米松组AQP1 mRNA相对吸光度比值分别为0.70±0.04、0.42±0.03、1.04±0.04,各组间差异有显著性(P<0.05);与空气组相比,高氧组AQP1 mRNA表达明显降低,高氧+地塞米松组AQP1 mRNA表达显著上调;AQP1蛋白表达与其mRNA变化一致.结论 高氧肺损伤时大鼠肺AQP1表达下调;地塞米松干预对肺损伤有保护作用,上调肺AQP1的表达可能是其作用机制之一.     

硫氧还蛋白及其还原酶在新生鼠高氧肺损伤中的表达

目的 研究硫氧还蛋白(Trx)及其还原酶(TR)在早产新生鼠高氧肺损伤的肺组织中的表达变化及意义.方法 早产新生SD大鼠,生后第一天随机分为空气组和高氧组.每组64只.两组分别于高氧或空气暴露后第4、7、10、14天提取肺组织总RNA,采取半定量逆转录聚合酶链反应(RT-PCR)测定Trx和TR mRNA表达;同时采用HE染色观察肺组织病理学变化,并进行辐射状肺泡计数(RAC).结果 病理学观察显示:与对照组比较,高氧组肺组织出现明显肺泡炎性改变和肺发育滞后,同时RAC值亦较空气组显著减少(P＜0.05).高氧暴露各组Trx、TR mRNA表达较空气组均明显增强(P＜0.05),且二者表达强度分别于第10天和第7天达高峰.结论 高氧上调肺组织Trx、TR表达;肺组织Trx系统表达增高可能在早产鼠高氧肺损伤的发病过程中发挥重要保护作用.     

机械通气在不同条件下对新生兔肺损伤及MMP-2表达的影响

目的 观察基质金属蛋白酶MMP-2在新生兔机械通气肺损伤中的表达和变化规律,研究肺损伤的分子机制.方法 将108只新生兔随机分配在高浓度氧、低浓度氧组,每组又分为高吸气峰压、中吸气峰压、低吸气峰压共6组进行机械通气并与正常未通气组6只进行对照,通气后1、3、6 h 3个时间点活杀取肺,病理切片MMP-2免疫组织化学染色,ELISA法检测肺组织匀浆中MMP-2的浓度,RT-PCR检测肺组织MMP-2mRNA的表达.结果 机械通气时,不同浓度的氧能使MMP-2表达上调,高浓度氧组为(5.38±1.57) μg/ml、低浓度氧组为(4.83±1.46) μg/ml,两组间差异无统计学意义,但同对照组(4.36±0.51) μg/ml相比,高氧组差异有统计学意义(F=3.658,P＜0.05).不同的机械拉伸力能使MMP-2上调,高、中、低压力3组分别为(5.10±1.79)、(5.35±1.38)、(4.96±1.49) μg/ml,3组间差异无统计学意义,但同对照组比,中压力组差异有统计学意义(F=2.764,P＜0.05).MMP-2同肺组织湿干比弱相关,同灌洗液细胞总数、细胞分类、肺损伤评分间统计学不相关.结论 新生兔机械通气6 h内,高浓度的氧通气能上调MMP-2,适中机械拉伸上调MMP-2,MMP-2同肺损伤有较少的相关性.     

硫氧还蛋白在早产鼠高氧肺损伤中的动态表达及其临床意义

目的研究高氧肺损伤早产鼠肺组织硫氧还蛋白(Trx)的表达及其临床意义。方法早产新生SD大鼠128只,生后d1随机分为空气组和高氧组,每组64只。分别于d1、4、7、10、14提取肺组织总RNA,采取半定量逆转录聚合酶链反应(RT-PCR)测定Trx mRNA水平;免疫组织化学方法检测肺组织切片凋亡蛋白半胱氨酸蛋白酶-3(Caspase-3);HE染色观察肺组织病理变化。结果高氧组肺发育明显受阻,与空气组比较,高氧暴露后Trx mRNA表达显著增强,且于d10时达高峰(P<0.05,0.01)。Caspase-3表达显著升高(P<0.05,0.01)。结论高氧上调早产鼠肺组织Trx mRNA的表达;肺组织Trx mRNA的表达增高可能在早产鼠高氧肺损伤的发生发展中发挥重要保护作用。     

p38丝裂素活化蛋白激酶在新生大鼠高氧肺损伤中的作用

目的 探讨p38丝裂素活化蛋白激酶（p38MAPK）在新生大鼠高氧肺损伤中的表达及其特异性抑制剂SB203580，对其产生保护作用的机制。方法 160只新生大鼠随机分为空气对照组、高氧肺损伤组、高氧肺损伤＋SB203580组和高氧肺损伤＋生理盐水组，建立模型。作用12、24、72h和1周后，分别处死大鼠。取右上肺进行肺组织病理学检查，取右下肺进行肺湿／干重比值（W／D）测定，取左肺以Western免疫印迹法检测肺组织中p38MAPK的表达，以酶联免疫吸附试验检测肺组织中TGF-β1的含量。结果 72h时，高氧肺损伤组和高氧肺损伤＋生理盐水组p38MAPK呈强阳性表达；在这两组中，肺组织TGF-β1浓度随时间延长呈持续上升趋势，在各时相点均明显高于空气对照组和高氧肺损伤＋SB203580组（P均〈0．01）。结论 p38MAPK参与了新生大鼠高氧肺损伤的过程，SB203580能够通过阻断p38MAPK表达，进而抑制TGF-β1表达来减轻这种损伤。     

新生大鼠缺氧缺血性脑损伤血红素氧化酶-1表达与脑细胞凋亡的关系

目的　探讨新生大鼠缺氧缺血性脑损伤 (HIBD)后海马区血红素氧化酶 1(HO 1)的表达 ,以及与细胞凋亡的关系。方法　 7日龄SD仔鼠 72只 ,随机分为HIBD组和假手术对照组。用原位杂交和免疫组化观测两组在不同时间点海马区HO 1mRNA及蛋白阳性细胞数量变化 ;用原位缺口末端标记 (TUNEL)法检测凋亡的脑细胞。结果　 1.HIBD组右侧海马HO 1mRNA表达 4h开始升高 (P <0 .0 5 ) ,12h达高峰 ,4 8h开始略有下降 ,72h后仍高于对照组 (P <0 .0 1)。 2 .HO 1蛋白表达与HO 1mRNA表达变化规律相一致。 3.凋亡检测发现 ,HIBD组右侧海马 12h凋亡细胞数增加 ,2 4h已有明显细胞凋亡 (P <0 .0 1) ,72h凋亡细胞减少 ,仍高于对照组 (P <0 .0 5 )。结论　新生大鼠HIBD后HO 1mRNA及其蛋白表达增加 ,可能参与神经细胞凋亡     

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.