柯萨奇病毒Ｂ３感染小鼠脑组织的钙超载及Ｎ—甲基—Ｄ—天冬氨酸受体Ｉ型mR

目的：为探索肠道病毒性脑炎的发病机制,以柯萨奇病毒（ＣＶ）B3感染小鼠,观察其神经细胞内钙离子[Ca^2 ]i及脑组织Ｎ－甲基－Ｄ－天冬氨酸受体Ｉ型（ＮＭＤＡＲ１）mRNA表达的变化,方法（１）取新生Ｂalb/c小鼠大脑皮质神经细胞,培养１２d 后,于CVB3感染12h（12份）用流式细胞仪检测细胞内[Ca^2 )]i,并用原子吸收光谱法检测神经细胞培养上清液中游离钙Ca^2 含量,（2)以CVB3颅内接种小,用NMDAR1的cDNA的cDNA地高辛标记探针对感染3d 后处死的石蜡包埋脑组织切片作原位杂交检测,结果：（1）感染12h小鼠脑神经细胞内[Ca^2 ]i值较对照组明显增高（P＜0．01）,感染组神经细胞培养上清液中Ca^2 含量低于对照组（P＜0．01）,（2）感染组脑组织中NMDAR1mRNA表达明显高于对照组（P＜0．01）,结论：CVB3感染小鼠脑神经细胞可导致细胞[Ca^2 ]i超载,且与Ca^2 内流有关。CVB3感染又可引起小鼠脑组织NMDAR1mRNA表达增高,其结果可能进一步引起Ca^2 内流和神经兴奋性毒性作用损伤脑组织。     

Bcl-2和Bax基因产物在小鼠柯萨奇病毒性心肌炎心肌组织中的表达

目的　探讨Bc1 2和Bax基因在病毒性心肌炎进展中的作用。方法　柯萨奇病毒B3 (CVB3 )感染12 5只雄性Balb/c小鼠 ,分别于d7、10、14、2 1、2 8随机取 2 0只实验组小鼠和 5只对照组小鼠处死 ,采用免疫组织化学方法检测CVB3 病毒性心肌炎小鼠不同时期心肌组织中Bcl 2和Bax蛋白表达情况。结果　病毒性心肌炎发病率为 86 % ,其中 80 %小鼠为轻、中度病变。正常对照组心肌中 ,无Bc1 2蛋白表达。在实验组中Bc1 2蛋白表达的动态变化与心肌病变积分呈显著正相关 (r =0 .93　P <0 .0 1) ,感染后 7～ 14d ,实验组小鼠心肌组织中Bax蛋白表达显著高于正常对照组 (P <0 .0 5 )。少数心肌浸润淋巴细胞表达Bc1 2和Bax。结论　Bc1 2和Bax基因参与病毒性心肌炎细胞凋亡的调控。     

卡托普利对病毒性心肌炎小鼠心肌线粒体结构及其酶活性的影响

目的　研究卡托普利对病毒性心肌炎小鼠心肌线粒体结构和三磷酸腺苷 (ATP)酶活性变化的影响。方法　将雄性Balb/c小鼠随机分为柯萨奇病毒B3 (CVB3 )感染组、CVB3 感染加卡托普利治疗组和对照组。以d3、d14为两个时相点 ,比较线粒体超微结构 ,线粒体Na+ K+ ATP酶、Ca2 + ATP酶活性变化。结果　感染组心肌细胞线粒体结构破坏 ,线粒体Na+ K+ ATP酶、Ca2 + ATP酶活性较对照组显著下降 (P均 <0 .0 1)。卡托普利治疗组各时相点线粒体结构破坏较轻 ,线粒体Na+ K+ATP 酶、Ca2 + ATP酶活性较感染组明显增高 (P均 <0 .0 5 )。结论　病毒性心肌炎早期即可见心肌线粒体结构破坏 ,ATP酶活性下降。卡托普利可有效保护心肌线粒体结构和功能     

胆红素诱导原代培养神经细胞内Ca2+含量的变化及MgSO4的拮抗作用

目的　探讨胆红素诱导原代培养神经细胞内Ca2 + 含量的变化 ,以及MgSO4 的拮抗作用。方法　采用胎鼠大脑皮层细胞作原代神经细胞培养 ,观察 10 0 μmol/L胆红素对不同成熟程度神经细胞内Ca2 + 含量的影响。原代培养神经细胞经Fura- 2 /AM负载 ,荧光图像分析测定细胞内Ca2 + 含量。结果　未成熟神经细胞 (培养2天 )暴露于 10 0 μmol/L胆红素 4h ,细胞内Ca2 + 从 75 .3nmol增加到 32 0 .8nmol;继续暴露 4h ,细胞内Ca2 + 增加到 40 0 .7nmol;去除胆红素暴露 ,细胞内Ca2 + 不能逆转。成熟神经细胞 (培养 8天 )暴露于 10 0 μmol/L胆红素 4h ,细胞内Ca2 + 从 70 .6nmol增加到 15 0 .8nmol;继续暴露 4h ,细胞内Ca2 + 不再增加 ;去除胆红素暴露 ,细胞内Ca2 + 浓度可显著降低。培养液中Mg2 + 加至 2mmol/L可减轻胆红素诱导的神经细胞内Ca2 + 含量变化。结论　胆红素可诱导原代培养的、未成熟的神经细胞Ca2 + 超载 ,MgSO4 可拮抗胆红素诱导的神经细胞Ca2 + 超载。     

胆红素诱导原代培养神经细胞内Ca2+含量的变化及MgSO4的拮抗作用

目的　探讨胆红素诱导原代培养神经细胞内Ca2 + 含量的变化 ,以及MgSO4 的拮抗作用。方法　采用胎鼠大脑皮层细胞作原代神经细胞培养 ,观察 10 0 μmol/L胆红素对不同成熟程度神经细胞内Ca2 + 含量的影响。原代培养神经细胞经Fura- 2 /AM负载 ,荧光图像分析测定细胞内Ca2 + 含量。结果　未成熟神经细胞 (培养2天 )暴露于 10 0 μmol/L胆红素 4h ,细胞内Ca2 + 从 75 .3nmol增加到 32 0 .8nmol;继续暴露 4h ,细胞内Ca2 + 增加到 40 0 .7nmol;去除胆红素暴露 ,细胞内Ca2 + 不能逆转。成熟神经细胞 (培养 8天 )暴露于 10 0 μmol/L胆红素 4h ,细胞内Ca2 + 从 70 .6nmol增加到 15 0 .8nmol;继续暴露 4h ,细胞内Ca2 + 不再增加 ;去除胆红素暴露 ,细胞内Ca2 + 浓度可显著降低。培养液中Mg2 + 加至 2mmol/L可减轻胆红素诱导的神经细胞内Ca2 + 含量变化。结论　胆红素可诱导原代培养的、未成熟的神经细胞Ca2 + 超载 ,MgSO4 可拮抗胆红素诱导的神经细胞Ca2 + 超载。     

三七皂甙治疗新生儿缺氧缺血性脑病的临床研究

目的：探讨三七皂甙对脑细胞的保护作用。方法：用新型钙染色剂Indo-1/Am检测缺氧缺血性脑病(HIE)患儿三七皂甙治疗前后细胞内钙变化,观察三七皂甙对细胞内钙超载的拮抗作用。将HIE患儿分为治疗组24例,对照组18例,同时设正常对照10例。在传统治疗的基础上,治疗组加用三七皂甙5～8 d,分别于入院时、治疗后24 h、72 h检测各组红细胞内游离钙(RBC[Ca2+]i)的浓度变化。结果：HIE治疗组及对照组RBC[Ca2+]i较正常足月儿升高,差异有非常显著性意义(P<0.01),HIE治疗组与HIE对照组比较,RBC[Ca2+]i在治疗前无明显差异(P>0.05),治疗后随着缺氧缺血状态的改善,检测48 h、72 h RBC[Ca2+]i,治疗组RBC[Ca2+]i明显下降,差异有显著性意义(P<0.01),HIE治疗组自身比较,RBC[Ca2+]i在治疗72 h后明显下降,差异有显著性意义[(2.619±0.175)vs(2.358±0.280);P<0.01]。治疗组中枢性呼吸衰竭、循环不良、胃肠功能紊乱治疗的有效率分别为100.0％(5/5)、83.3％(20/24)、91.7％(22/24),对照组有效率分别为20.0％(1/5)、33.3％(6/18)、16.7％(3/18),(P<0.01)。结论：三七皂甙能缓解细胞内钙超载,保护脑组织,改善临床症状。     

盐酸戊乙奎醚对新生大鼠缺氧缺血性脑病模型脑细胞钙离子浓度及天冬氨酸特异性半胱氨酸蛋白酶-3表达的影响

目的 探讨盐酸戊乙奎醚(长托宁)对新生大鼠缺氧缺血性脑病( hypoxic-ischemic encephalopathy,HIE)模型脑细胞内钙离子浓度[Ca2+]i及天冬氨酸特异性半胱氨酸蛋白酶-3(cysteinecontaining aspartate-specific proteases-3,Caspase-3)表达的影响.方法 新生7日龄健康SD大鼠,共128只,随机分为4组:假手术对照组(Sham组,n=32),HIE模型组(HIE组,n=32),尼莫地平干预组(N组,n=32)及长托宁干预组(P组,n=32).各组于手术后不同时间点(2h,4h,6h,12h)分批采集脑组织标本.用共聚焦激光显微镜和荧光显微镜分别观察不同组及各组不同时间点大鼠脑皮层细胞[ca2+]i和Caspase-3的表达.结果 HIE组大鼠脑皮层细胞[Ca2+]i和Caspase-3的表达,显著高于Sham组(P＜0.01),缺氧缺血后2h,[Ca2+]i和Caspase-3表达即增加,随着时间延长,[Ca2+]i和Caspase-3表达逐渐增加,12 h时显著高于其他时间点(P＜0.05).N组与P组的[Ca2+]i和Caspase-3 表达较HIE组明显降低(P＜0.01),但仍高于Sham组(P＜0.01).N组与P组之间[Ca2+]i和Caspase-3表达差异无统计学意义(P＞0.05).N组与P组内12h时[Ca2+]i和Caspase-3表达与6h比较,差异无统计学意义(P＞0.05).结论 长托宁可显著缓解缺氧缺血后脑皮层细胞钙超载并减少Caspase-3表达,进而减少脑细胞死亡,起到保护脑细胞的作用.     

黄芪甲甙对病毒性心肌炎小鼠神经生长因子表达的影响

目的探讨黄芪活性成分黄芪甲甙对病毒性心肌炎(VM)小鼠心肌中神经生长因子(NGF)表达的影响。方法Balb/c小鼠100只,随机分成6组。非感染小鼠腹腔无菌注射病毒培养液,分为正常对照组[A组,10只,以羧甲基纤维素钠(CMC)0.1 mL灌胃7 d]、高剂量对照组(B组,10只,9%黄芪甲甙液0.1 mL灌胃7 d);余80只小鼠以柯萨奇病毒(CVB3)腹腔无菌注射制作VM模型,VM小鼠随机分为心肌炎对照组和低、中、高剂量干预组,分别以CMC、1%、3%、9%黄芪甲甙液0.1 mL灌胃7 d(分别为C、D、E、F组,每组20只)。14 d后处死小鼠并取其心脏,采用RT-PCR和免疫组织化学检测NGF mRNA及蛋白表达水平。结果F组小鼠较C组死亡率明显降低,B组无小鼠死亡;与C组比较,NGF mRNA、蛋白表达水平以及心肌病变积分在F组均显著下降,而低、中剂量黄芪甲甙对心肌炎小鼠NGF mRNA、蛋白表达及心肌病变积分无明显影响。结论黄芪甲甙可能通过抑制NGF表达,对Balb/c小鼠CVB3心肌炎具有良好的治疗作用。     

大剂量维生素C对病毒性心肌炎小鼠心肌腺苷酸酶活性和钙代谢的影响

有研究显示 ,心肌感染病毒后Ｃａ2 内流增加[1 ] ,维生素Ｃ(ＶｉｔＣ)能明显降低病毒滴度、增强心肌活力、减少细胞死亡[2 ] 。但ＶｉｔＣ能否改善病毒感染后心肌Ｃａ2 内流、减轻细胞Ｃａ2 超载 ,尚无相关文献报道。本实验通过复制小鼠病毒性心肌炎模型 ,对此进行探讨。对象6～ 8周龄近交系雄性Ｂａｌｂ ｃ小鼠 70只 ,体重 (2 0± 2 )ｇ。随机分为柯萨奇Ｂ3病毒 (ＣＶＢ3)感染组 (ＩＧ)、ＣＶＢ3感染加大剂量ＶｉｔＣ治疗组 (ＩＶＧ)及对照组 (ＣＧ)。ＩＧ及ＩＶＧ均分设感染后第 3天 (6只 )、14天 (11只 )及 30天 (13只 ) 3个时相点…     

骨膜蛋白在实验性病毒性心肌炎小鼠的表达及意义

目的 探讨骨膜蛋白(periostin)在病毒性心肌炎(VMC)中的表达及可能作用.方法 应用柯萨奇病毒B3(CVB3)感染4周龄Balb/c小鼠,建立VMC小鼠模型.100只小鼠分为正常对照组(n ＝ 40)和病毒组(n ＝ 60).在CVB3感染后第0、7、14、28、56天留取标本,测定各实验组不同时间点心肌胶原容积分数(CVF)值(Masson染色)、血清血管紧张素(Ang)Ⅱ水平(ELISA法)、心肌组织中骨膜蛋白和转化生长因子(TGF)β1 mRNA相对表达量(RT-PCR).将骨膜蛋白 mRNA相对表达量与其他指标进行直线相关性分析.结果 VMC组小鼠骨膜蛋白mRNA表达量从病毒感染后第7天开始增加,第28天持续升高,第56天时表达量最高(P ＜ 0.01).骨膜蛋白mRNA表达量与CVF、血清AngⅡ、TGFβ1 mRNA相对表达量均呈直线正相关(r ＝ 0.870 ～ 0.943,P均＜ 0.01).结论 骨膜蛋白的异常表达在VMC的发生、发展中起重要作用,可能参与了VMC心肌纤维化的发生过程.同时骨膜蛋白与AngⅡ及TGF-β1表达相关,可能成为抗VMC心肌纤维化的新靶点.     

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.     

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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.     

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   

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.     

Overcoming surfactant inhibition with polymers

Inhibition of the function of pulmonary surfactant in the alveolar space is an important element of the pathophysiology of many lung diseases, including meconium aspiration syndrome, pneumonia and acute respiratory distress syndrome. The known mechanisms by which surfactant dysfunction occurs are (a) competitive inhibition of phospholipid entry into the surface monolayer (e.g. by plasma proteins), and (b) infiltration and destabilization of the surface film by extraneous lipids (e.g. meconium-derived free fatty acids). Recent data suggest that addition of non-ionic polymers such as dextran and polyethylene glycol to surfactant mixtures may significantly improve resistance to inhibition. Polymers have been found to neutralize the effects of several different inhibitors, and can produce near-complete restoration of surfactant function. The anti-inhibitory properties of polymers, and their possible role as an adjunct to surfactant therapy, deserve further exploration.     

Understanding infant formula

The World Health organisation recommends breast feeding infants for the first six months of life. When this breast feeding does not occur either through parental choice or medical need, infant formulas will be required. There is a bewildering array of formulas on the UK market for many different requirements. When faced with an unsettled infant many parents (and healthcare professionals) will experiment with the infant formula available and then attend the paediatric clinic looking for help and advice. It is therefore essential that paediatricians understand what milks are available and what the key differences between different products are. This review attempts to provide a simple guide through many of the formulations currently available in the UK; and offers advice for the dietary management of the child with extra calorie requirements, infants with cow's milk protein allergy, gastro oesophageal reflux disease, apparent unresolved hunger and infantile colic. Whatever the underlying condition, there is likely to be an infant formula that is suitable in this generation of ever expanding formulations.