缺锌对哮喘大鼠Th1/Th2型细胞因子的影响

目的 探讨缺锌对哮喘大鼠Th1／Th2型细胞因子干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）的影响。方法 建立缺锌及哮喘模型，SD大鼠32只，体质量45～65g，按体质量随机分为4组，每组8只。A组为缺锌饲料＋卵清蛋白（OVA）激发组；B组为正常锌饲料＋OVA激发组；C组为正常锌饲料配对饲养＋OVA激发组；D组为正常锌饲料＋生理盐水激发组。利用酶联免疫吸附法（ELISA）检测肺组织IFN-γ和IL-4水平；逆转录PCR（RT-PCR）检测肺组织IFN-γ、IL-4mRNA表达。结果 与D组比较，A～C组大鼠肺组织匀浆IFN-γ水平、IFN-γ mRNA表达均明显减少（P均〈0．05），而IL-4水平、IL-4mRNA表达均明显增加（P均〈0．05）。与B组比较，A组大鼠IFN-γ水平、IFN-γmRNA明显减少（P均〈0．05）；IL-4水平无显著性差异（P〉0．05），IFN-γ／IL-4比例明显减少（P〈0．05）。B与C组比较检测结果均无显著性差异。结论 哮喘大鼠缺锌时其IFN-γ明显减少，而IL-4不受影响，Th1／Th2细胞比例严重失衡，是缺锌时呼吸道炎症反应增加原因之一。     

IL-12对小鼠哮喘模型Th亚群和气道炎症的影响

目的观察腹腔注射IL-12对哮喘小鼠Th亚群的调节和气道炎症的影响,并寻找更有效的干预阶段。方法将36只BALB/c小鼠分为4组,每组9只,分别为哮喘组、IL-12干预A组、IL-12干预B组及正常对照组。采用卵白蛋白(OVA)腹腔注射致敏加雾化吸入激发的方法制成哮喘模型,正常对照组小鼠同时以等量的生理盐水腹腔注射加雾化吸入。干预A组在致敏和激发阶段均给予IL-12腹腔注射进行干预,干预B组仅在激发阶段给予IL-12腹腔注射进行干预。最后1次激发结束后24h处死所有小鼠,取左肺作病理切片(HE染色),右肺提取总RNA并逆转录成cDNA后,应用半定量RT-PCR法检测肺组织内IFN-γ基因mRNA的转录表达情况,实时定量PCR法检测肺组织内IL-4基因mRNA的转录表达情况。结果与正常对照组相比,哮喘组小鼠肺组织内IFN-γmRNA表达显著减少(P<0.05),IL-4mRNA表达显著增多(P<0.05);与哮喘组相比,IL-12干预A、B组小鼠肺组织内IFN-γmRNA表达均显著增多(P均<0.05),IL-4mRNA表达均显著减少(P均<0.05)。干预A组与干预B组相比,IL-4mRNA的表达水平差异有统计学意义(P<0.05),而IFN-γmRNA表达水平差异无统计学意义(P>0.05)。结论IL-12能有效纠正哮喘小鼠失衡的Th亚群及控制气道炎症;致敏、激发阶段同时干预与仅激发阶段的干预均能有效的抑制Th2型免疫反应和气道炎症改变,前者能更有效的降低IL-4mRNA的表达水平。     

卡介苗新生期接种和呼吸道合胞病毒感染对小鼠实验性哮喘的联合作用

目的观察卡介苗（BCG）新生期接种和呼吸道合胞病毒（RSV）感染对小鼠实验性哮喘的联合作用.方法新生期小鼠分5组：对照组、鸡卵白蛋白（OVA）组、BCG＋OVA组、RSV＋OVA组、BCG＋RSV＋OVA组,除对照组外,其他各组为哮喘组都进行OVA致敏和激发.最后一次激发后24 h内测气道高反应性,进行支气管肺泡灌洗,观察支气管肺泡灌洗液（BALF）中白细胞数及分类,ELISA方法检测肺泡灌洗液中细胞因子含量和血清中OVA特异性IgE水平,肺组织HE染色进行炎症病理学分析.结果（1）各哮喘组BALF中白细胞总数及各类炎症细胞数都比对照组显著升高.BCG＋OVA组的白细胞总数、嗜酸性粒细胞和淋巴细胞计数显著低于其他哮喘组（P＜0.05或0.01）.（2）各哮喘组与对照组比较BALF中IL-4水平显著增高（P＜0.05）,IFN-γ水平显著减低（P＜0.05）,4组哮喘组之间两两比较差异无统计学意义.各实验组IL-10水平差异无统计学意义.（3）各哮喘组血清中OVA特异性IgE水平都比对照组明显增高（P＜0.05或0.01）,各哮喘组之间比较差异无统计学意义.（4）随吸入乙酰甲胆碱浓度增加各组气道反应性明显增加.RSV＋OVA组和BCG＋RSV＋OVA组的气道反应性最高,其后依次是OVA组、BCG＋OVA组、对照组.RSV＋OVA组和BCG＋RSV＋OVA组间比较差异无统计学意义.（5）各哮喘组的炎症评分都显著高于对照组.BCG＋OVA组的细支气管周围炎症和支气管周围嗜酸性粒细胞浸润百分率显著低于另外三组（P均＜0.05）,肺泡炎症反应明显低于OVA组和BCG＋RSV＋OVA组（P均＜0.05）.结论新生期BCG接种在OVA致敏/激发的小鼠哮喘模型中能减轻哮喘炎症和气道高反应性,但这种抗哮喘作用可被RSV感染逆转.     

哮喘小鼠肺组织GATA-3及Th2型细胞因子表达的意义

目的探讨Th2特异性转录因子GATA-3及Th2型细胞因子（IL-5、13）在支气管哮喘（哮喘）小鼠中的表达及意义。方法BALB/c小鼠16只随机分为对照组和模型组。卵清蛋白致敏制备哮喘模型。HE染色观察小鼠肺组织病理变化,逆转录聚合酶链反应（RT-PCR）法检测小鼠肺组织GATA-3 mRNA表达,酶联免疫吸附法（ELISA）检测小鼠肺组织IL-5、-13蛋白的表达。结果HE染色示模型组支气管周围见大量炎性细胞浸润,而对照组则不明显。与对照组相比,模型组GATA-3 mRNA和IL-13蛋白表达量明显增加（Pa〈0.05）。结论哮喘小鼠肺部存在GATA-3高表达,GATA-3可直接调控IL-5、-13表达,参与哮喘呼吸道炎症的发生。     

血管活性肠肽对哮喘小鼠气道炎症及Th17/Treg平衡的影响

目的探讨血管活性肠肽(VIP)对哮喘小鼠气道炎症的影响,以及对Th17细胞/调节性T细胞(Treg)失衡的调控作用。方法将30只BALB/c小鼠随机分成对照组、哮喘组、VIP组,每组10只。利用卵白蛋白(OVA)致敏和激发制作急性哮喘小鼠模型;对照组致敏和激发阶段均以生理盐水代替OVA;VIP组每次OVA激发前,先以VIP溶液(20μg/m L)雾化吸入30 min。留取各组小鼠支气管肺泡灌洗液、肺组织等标本。利用苏木精-伊红染色观察肺组织病理变化;ELISA法检测肺泡灌洗液中Th17/Treg相关细胞因子水平;免疫组化及Real-time PCR检测肺组织中Th17细胞特异性转录因子RORγt及Treg特异性转录因子Foxp3表达情况。结果病理组织学结果显示VIP组小鼠肺组织气道炎症表现较哮喘组减轻。哮喘组小鼠BALF中IL-17浓度高于对照组(P0.01),VIP组IL-17浓度较哮喘组降低(P0.01),但仍高于对照组(P0.01)。哮喘组BALF中IL-10浓度低于对照组(P0.01),VIP组IL-10浓度较哮喘组升高(P0.01),但仍低于对照组(P0.01)。哮喘组小鼠肺组织RORγt mRNA及蛋白表达高于对照组(P0.01),Foxp3 mRNA及蛋白表达低于对照组(P0.01);VIP组肺组织RORγt mRNA及蛋白表达水平低于哮喘组(P0.01),Foxp3 mRNA及蛋白表达水平高于哮喘组(P0.05)。结论哮喘小鼠存在Th17/Treg免疫失衡,VIP可通过调控Th17/Treg免疫失衡而改善哮喘小鼠气道炎症。     

树突状细胞对呼吸道合胞病毒感染T细胞免疫反应的调节作用

目的研究在呼吸道合胞病毒（RSV）感染过程中树突状细胞（dendritic cell,DC）的数量和功能变化,探讨其对T细胞免疫反应的调节作用。方法BALB／c小鼠感染RSV后,分离并用流式细胞仪检测肺DC的数量和成熟标志;分离肺淋巴细胞并在体外培养,ELISA测定培养上清中Th2和Th1细胞因子的水平;在体外RSV或流感病毒（IFZ）感染骨髓衍生树突状细胞（BMDC）48h,检测培养体系上清中IL-12、IL-10水平。在BMDC和T淋巴细胞共培养体系中加入重组IL—10,观察对T细胞分泌细胞因子的影响。结果RSV感染小鼠肺淋巴细胞在体外培养情况下IL—13、IL-4和IL-5分泌显著增加,均高于IFZ感染小鼠（P均〈0．01）;IFN-γ分泌水平在RSV或IFZ感染小鼠均升高,但两者间差异无统计学意义（P〉0．05）。RSV感染小鼠后3d,肺DC数量增多,DC成熟标记分子MHCⅡ-Ⅰa、CD86和CDS0表达增加（P均〈0．01）;IFZ感染BMDC48h,培养上清中IL—10显著增高（P〈0．01）,而在RSV感染组不升高（P〉0．05）。在BMDC和T淋巴细胞共培养体系中加入重组IL—10后,IFN-γ的分泌水平显著增加,IL-13和IL-5的分泌水平受到抑制（P均〈0．01）。结论RSV感染过程中,DC数量增加,成熟度提高,但分泌IL-10不足,通过补充外源性IL-10实验证明这是RSV感染引起Th2反应为主的原因之一。     

食物过敏动物模型中辅助性T淋巴细胞1/辅助性T淋巴细胞2相关细胞因子变化

目的观察食物过敏动物模型辅助性T淋巴细胞(Th)1/Th2型细胞因子变化,探讨其在食物过敏发生中作用。方法选取110只Balb/c小鼠,用卵清蛋白(OVA)致敏、激发建立食物过敏实验性小鼠模型,设立对照组及致敏组,进行食物过敏原皮肤点刺及小肠病理组织学检测,ELISA检测其血清特异性IgE、IL-4、IFN-γ水平,real-tim e PCR检测其小肠IL-10、TGF-βmRNA表达。结果致敏组小鼠IL-4水平较对照组升高、IFN-γ水平较对照组降低,均有统计学意义(Pa<0.05);其小肠IL-10、TGF-βmRNA表达较对照组降低,有统计学意义(Pa<0.05)。结论食物过敏是以Th2型淋巴细胞占优势的过敏性疾病;Th1/Th2细胞因子失平衡可能参与食物过敏发生,并可能在其中发挥重要作用。     

GATA-3在支气管哮喘小鼠肺组织表达及地塞米松对其抑制作用

目的探讨转录因子GATA-3在支气管哮喘小鼠肺组织中表达及地塞米松（Dex）对其干预作用。方法采用卵清蛋白（OVA）致敏方法建立支气管哮喘小鼠模型;Balb/c小鼠24只随机分为对照组、哮喘组、Dex治疗组各8只。采用免疫组织化学方法检测肺组织GATA-3蛋白表达;反转录聚合酶链反应（RT-PCR）方法检测肺组织GATA-3 mRNA相对水平;流式细胞技术检测小鼠脾CD4 T细胞白细胞介素-4（IL-4）水平;HE染色评价各组小鼠气管炎性反应变化。结果哮喘组小鼠肺组织GATA-3及mRNA水平、IL-4水平均明显高于对照组（P〈0.05）,Dex治疗组GATA-3阳性细胞数及mRNA水平、IL-4水平均明显低于哮喘组,具有显著性差异（P〈0.05）。HE染色结果表明Dex治疗组气管炎性反应明显缓解。结论GATA-3在支气管哮喘小鼠肺组织呈高表达。Dex可阻断哮喘鼠肺组织GATA-3表达,抑制Th2型细胞因子释放,减轻气管炎性反应。     

新生期小鼠卡介苗接种和呼吸道合胞病毒感染对肺部炎症和免疫状态的影响

目的研究小鼠新生期卡介苗（BCG）接种对呼吸道合胞病毒（RSV）感染后肺部炎症和免疫状态的影响。方法新生BALB／c小鼠分4组：A组为对照组、B组为BCG接种组、C组为RSV感染组、D组为BCG接种后RSV感染组。RSV感染后1周观察肺泡灌洗液（BALF）中白细胞数及分类，ELISA法检测BALF中IFN-γ、IL-4、IL-10含量，RT-PCR检测肺组织Toll样受体4（TLR4）mRNA表达，HE染色进行肺组织炎症病理评分。结果C、D两组BALF中自细胞总数、淋巴细胞数、中性粒细胞数、IFN．7含量以及肺组织TLR4 mRNA表达量和炎症病理评分显著高于A、B组（P〈0．01或0．05），BALF中IL-4、IL-10含量4组间差异无显著性，C、D两组之间在各项指标上差异无显著性。结论小鼠新生期BCG接种对RSV感染诱发的肺组织炎症和免疫状态无影响。     

呼吸道合胞病毒感染大鼠转录因子T-bet表达及与Th1/Th2的关系

目的 探讨呼吸道合胞病毒(RSV)感染时肺T-bet(T-box expressed in T cells)表达对Th1/Th2类细胞因子分泌的调控作用.方法 随机将20只SD大鼠分为对照组、RSV感染组,每组10只.在第6周时测定气道反应性;取肺组织苏木素伊红(HE)染色观察病理改变;通过蛋白质印迹法检测T-bet在肺组织中的表达;酶联免疫吸附法检测肺组织匀浆白细胞介素4(IL-4)和干扰素Ⅱ型(IFN-γ)水平.结果 RSV感染组大鼠肺部病理学切片呈现典型的间质性肺炎表现,而且RSV感染组大鼠气道阻力程度明显高于对照组.肺组织中T-bet表达RSV感染组相对灰度值0.61 ± 0.23较对照组0.81 ± 0.35显著减低(t ＝ 16.345,P ＜ 0.01).RSV感染组的IL-4蛋白水平较对照组明显增高(P ＜ 0.01),而IFN-γ蛋白水平明显减低(P ＜ 0.01).肺T-bet蛋白水平与IFN-γ蛋白水平呈正相关(r ＝ 0.725,P ＜ 0.01),与IL-4 蛋白水平及IL-4/IFN-γ比值均呈负相关(r ＝ -0.837、-0.864,P均＜ 0.01).结论 RSV感染可以下调T-bet在肺组织中表达,并与Th1/Th2免疫失衡相关.     

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.