转录因子T-bet、GATA-3在过敏性紫癜患儿发病机制中的作用

目的 探讨转录因子T-bet和GATA-3在过敏性紫癜(HSP)患儿发病机制中的作用.方法 本院儿科2009年2月-2010年2月收治急性期HSP患儿46例(HSP组)以及健康对照儿童30例(健康对照组).采用SYBR Green I实时荧光定量PCR方法检测其外周血单个核细胞T-bet mRNA、GATA-3 mRNA的表达.应用Luminex-100液相芯片技术检测其血浆细胞因子IL-2、IL-4、IFN-γ水平.结果 HSP组患儿外周血单个核细胞T-bet mRNA相对表达水平(53.98±35.79)低于健康对照组(181.56±96.90)(P＜0.001).GATA-3 mRNA相对表达水平(964.30±655.18)高于健康对照组(78.09±57.20)(P ＜0.001).HSP组血浆细胞因子IL-2[( 16.54±7.38) ng?L-1],IFN-γ[(11.31±8.02) ng?L-1]的表达显著低于健康对照组[(35.73±22.66) ng?L-1,(30.67±21.29) ng?L-1](P＜0.001,0.05),IL-4[(74.66±26.15) ng.L-1]的表达高于健康对照组[(51.81±27.76) ng?L-1].IFN-γ水平与T-bet mRNA表达呈正相关(r=0.882,P＜0.01);IL4水平与GATA-3 mRNA的表达呈正相关(r=0.886,P＜0.01).结论 HSP患儿急性期存在Th1/Th2失衡,主要表现为Th2优势活化,该失衡与其特异性转录因子T-bet mRNA表达降低、GATA-3 mRNA 表达增高有关,本研究为阐明Th1/Th2失衡的分子机制之一提供了实验依据.     

哮喘缓解期患儿Th1/Th2相关上游转录因子及细胞因子的表达

目的从基因和蛋白质水平了解哮喘缓解期患儿T辅助淋巴细胞(Th)免疫平衡状况。方法选取24例哮喘缓解期和12例健康儿童作为对照,采用酶联免疫吸附法(ELISA)和实时荧光定量PCR(real time PCR)法分别测定外周血细胞因子干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)水平及外周血单个核细胞(PBMC)中特异性转录因子T-bet、GATA-3表达情况。结果病例组血浆IL-4水平(37.14±12.77)ng/L显著高于对照组(27.08±6.12)ng/L,t=2.467 P<0.05;病例组PBMC中GATA-3的表达强度(5.45±1.11)较对照组(4.70±0.86)显著增高(t=2.049 P<0.05);病例组与对照组比较IFN-γ/IL-4及T-bet/GATA-3比率均显著降低(t=-2.208,-4.212 P<0.05,P<0.01)。结论哮喘缓解期患儿体内存在基因和蛋白质水平Th1/Th2平衡的紊乱,以Th2亚群亢进为主。     

急性毛细支气管炎患儿血清白细胞介素13、γ-干扰素表达的意义

目的 探讨急性毛细支气管炎患儿血清IL-13、IFN-γ表达及其临床意义.方法 用酶联免疫吸附法(ELISA)检测42例急性期毛细支气管炎患儿(其中轻症组22例,重症组20例)和20名健康婴儿血清IL-13、IFN-γ水平.采用方差分析和成组t检验,检测各组间差异.结果 1.急性毛细支气管炎患儿血清IL-13[(6.88±2.12 )ng/L]明显高于对照组[(5.48±1.28 )ng/L](P＜0.05).2.急性期毛细支气管炎患儿IFN-γ[(10.71±2.44 )ng/L]明显高于对照组[(9.20±1.54)ng/L](P＜0.05);轻症组明显高于重症和对照组(P＜0.05),而重症与对照组则无显著性差异(P＞0.05).结论 1.IL-13参与毛细支气管炎的发病过程,但其水平不能反映病情严重程度;2.IFN-γ水平在轻症毛细支气管炎组明显增高,而重症组不增高,可能与急性重症毛细支气管炎患儿IFN-γ产生受抑制有关.     

ITP患儿外周血T细胞T-bet/GATA-3基因的检测及意义

目的研究急慢性ITP患儿Th1、Th2类细胞因子和转录因子T-bet和GATA-3的相关性,探讨其在ITP发病机制。方法选择30例ITP患儿,运用T细胞分离富聚柱得纯化的T细胞,分别用ELISA法和RT-PCR技术检测ITP患儿血清中的IFN-γ、IL-4水平和外周血T细胞转录因子T-bet和GATA-3的表达状态,设30例健康儿童为对照组。结果慢性ITP患儿血清中IL-4水平较正常人降低,IFN-γ的水平升高,GATA-3 mRNA表达水平下降,T-be tmRNA表达水平升高;而急性ITP患儿外周血中IL-4水平较正常人升高,血清中IFN-γ水平及T-bet/GATA-3比值无显著性差异。结论慢性ITP患儿外周血Th1/Th2比例失衡,T-bet/GATA-3的表达失调,可能引起慢性ITP的免疫紊乱:而急性ITP患儿的发病机制可能与T-bet/GATA-3的表达无关。     

重症肌无力被动转移幼鼠的黏膜免疫耐受研究

目的 观察特异性耐受原-双类似物(Lys262-Ala207)对重症肌无力被动转移(PTMG)小鼠模型的黏膜免疫效果,探讨其作用机理及该方法在模型应用中的可行性.方法 将 C57BL/6幼年雌性小鼠60只分为3组:耐受组、模型组、对照组,各20只.耐受组和模型组建立PTMG模型.耐受组在致敏前10 d,每天经鼻腔滴人含耐受肽-双类似物25μg的PBS 50μl;模型组连续10 d经鼻腔滴入不含耐受肽的PBS 50μl;正常对照组不做特殊处理.观察实验各组小鼠的体重、临床评分、白细胞介素4(IL-4)、γ干扰素(IFN-γ)、TGF-a1及其他各项指标的改变情况.结果 模型组小鼠表现较为典型的PTMG症状;耐受组小鼠经耐受治疗有效,其临床症状较轻.血清乙酰胆碱受体抗体(AChRAb)含量耐受组为(16.01±1.09)mg/L,模型组为(28.12±1.28)mg/L,均高于对照组[(1.60±0.28)mg/L](t=44.37,70.27,P<0.01).耐受组外周血中TGF-B1[(437.19±1.93)ng/L]高于模型组[(175.63±3.12)ng/L](t=36.07,P<0.01),模型组IL-4、IFN-γ[(193.37±3.95)ng/L,(320.46±2.14)ng/L],均高于耐受组[(141.02±3.11)ng/L,(187.99±4.67)ng/L](t=37.20、51.69,P<0.01).各因子与对照组比较仍未达正常水平,差异有统计学意义(t=26.65、31.05、49.02,P<0.01).结论 双类似物鼻黏膜耐受对缓解PTMG有效,表现为:血清AChRAb含量降低,外周血中TGF-a1分泌增高,IL-4、IFN-γ分泌降低和临床症状的缓解等.同时该方法还具有使用方便、安全等特点.     

T-bet基因传递对支气管哮喘模型小鼠T淋巴细胞亚群的免疫调节作用

目的 应用携带T-bet基因的重组腺相关病毒载体(rAAV-T-bet)经鼻干预支气管哮喘(哮喘)小鼠模型,探讨T-bet基因传递对哮喘小鼠T淋巴细胞亚群的免疫调节及改善炎症的作用.方法 采用随机数字表法将40只健康的6～8周龄SPF级Balb/c小鼠随机分为正常对照组(A组)、哮喘模型组(B组)、rAAV介导的增强型绿色荧光蛋白(rAAV-eGFP)干预对照组(C组)及rAAV-T-bet干预组(D组),每组10只.B、C、D组以卵清蛋白(OVA)致敏和激发,建立哮喘小鼠模型,其中C、D组在第18、19天分别经鼻滴入等剂量rAAV-eGFP、rAAV-T-bet进行干预;B组处理方法同上,以等剂量的9 g/L盐水干预.于最后一次激发24 h后处死小鼠并取材.应用免疫组织化学方法观察小鼠肺组织中转录因子T-bet、GATA-3和Foxp3水平;计数BALF中细胞总数并分析其细胞类型;ELISA法测定BALF中细胞因子IFN-γ、IL-4、IL-5表达水平.结果1.B组、C组中T-bet表达细胞的阳性率均明显低于A组(P均＜0.05),GATA-3表达细胞的阳性率明显高于A组(P均＜0.05),D组中T-bet表达明显增强,GATA-3表达则显著下降(P均＜0.05),Foxp3的表达阳性率在各组间的差异与T-bet的表达类似.2.D组BALF中细胞总数为(0.35±0.11)×109/L,其中EOS比例为0.04±0.02,虽高于A组,但明显低于B、C组(P均＜0.05);IL-4和IL-5分别为(158±55) ng/L、(68±22) ng/L,显著低于B、C组(P均＜0.05),高于A组但差异无统计学意义(P均＞0.05),而IFL-γ水平[(113±35) ng/L]明显低于B、C组(P均＜0.05),与A组比较水平相近(P＞0.05).结论 在建立小鼠哮喘模型的基础上,经鼻应用rAAV-T-bet干预可使T-bet基因顺利定向插入到受转染细胞的基因组中,成功编码T-bet蛋白,并进一步对哮喘小鼠模型发挥相应的免疫调节作用.     

毛细支气管炎患儿免疫功能状态评价的意义

目的探讨毛细支气管炎(毛支)患儿的免疫功能状态的意义。方法采用酶联免疫吸附分析法(ELISA)和逆转录聚合酶链反应(RT-PCR)测定25例毛支患儿外周血单个核细胞体外产生IL-12、18、10、4,IFN-γ及血浆IgE的水平及膜表面蛋白分子淋巴细胞活化基因3(LAG-3)mRNA的表达。门诊体检的健康儿童14例作为正常对照组。结果毛支患儿急性期IL-12 (390．94±95．88)ng／L及II-18(146．23±55 81)ng／L水平明显低于正常对照组[(452．44±195．07)、(246．69±121．29)ng／L] (P<0．05),IL-10(302．40±100．04)ng／L、IL-4(30．52±17．65)ng／L及IgE(82．90±51．4)ng／L水平明显升高(P分别<0．05, 0．01,0．05),IFN-γ(103．25±82．15)ng／L水平虽低于正常对照组,但无显著差异(P>0．05)。IFN-γ/IL-4比率(3．6±0．9:1．0) 较对照组(9．7±2．1:1．0)降低约3倍(P<0．05)。TH1细胞表面标记LAG-3 mRNA的表达明显低(P<0．05)。结论毛支患儿存在TH1/TH2失衡、TH1功能下降、TH2功能占优势的状态,与APC的功能状态及其所分泌的上游调控细胞因子(IL-12、18、 10)的平衡紊乱有关。     

川崎病患儿急性期血浆IFN-γ、IL-4水平变化研究

目的探讨川崎病(KD)急性期T细胞的功能状态及临床意义。方法ELISA法检测急性期KD患儿35例和10例健康儿童血浆Th1/Th2细胞的代表性细胞因子IFN-γ、IL-4水平变化,评价T细胞功能状态。结果急性期KD患儿血浆[IFN-γ(34±17)ng/L、IL-4(45±23)ng/L]水平较对照组[IFN-γ(59±21)ng/L、IL-4(69±28)ng/L]均明显下降,差异有显著性(P<0.05);两组IFN-γ/IL-4值比较,差异无显著性(P<0.05)。冠状动脉受累(CA)组[IFN-γ(28±15)ng/L、IL-4(31±14)ng/L]与无冠状动脉受累(NCA)组[IFN-γ(39±25)ng/L、IL-4(43±21)ng/L]比较,差异无显著性(P>0.05);同样,两组IFN-γ/IL-4值比较,差异无显著性(P>0.05)。结论KD急性期T细胞功能受抑,这或许是KD免疫学特征之一。     

儿童过敏性哮喘尘螨特异性免疫治疗中干扰素-γ白介素4 T-bet和GATA-3 mRNA表达变化的研究

目的 探讨在过敏性哮喘尘螨特异性免疫治疗(SIT)过程中,与TH1/TH2细胞分化相关的干扰素-γ(IFN-γ)、白介素4(IL-4)、T-het和GATA-3 mRNA在外周血单个核细胞(PBMCs)中表达的变化.方法 根据进行抗原特异性免疫治疗(SIT)的时间,采集郑州大学第一附属医院儿科门诊2007年11月至2008年9月收治的过敏性哮喘惠儿36例,对其SIT治疗前、治疗16周后和治疗1年后的外周静脉血分离外周血单个核细胞,提取总RNA,逆转录成cDNA,利用SYBR Green Ⅰ荧光定量PCR的方法检测每组中IFN-γ、IL-4、T-bet和GATA-3mRNA表达情况.同时,观察患儿治疗前后的临床疗效.结果 随着SIT治疗的进行,患儿临床症状明显减轻,外周血中在mRNA表达水平IFN-γ/IL-4和T-bet/GATA-3的比值逐渐升高.结论 哮喘患儿特异性免疫治疗有效;治疗后TH1/TH2细胞的失衡得到一定的纠正;T-bet和GATA-3转录因子在哮喘的发生和SIT治疗中起重要作用.     

白介素-13与γ-干扰素在急性毛细支气管炎患儿血清中的表达及其临床意义

目的探讨白介素-13(IL-13)、γ-干扰素(IFN-γ)在急性期毛细支气管炎患儿血清中的表达及与病情轻重之间的相关性。方法用酶联免疫吸附(ELISA)法测定2005-02—2005-11于遵义医学院附属医院治疗的42例急性期毛细支气管炎患儿(其中轻症组22例,重症组20例)和16名健康婴儿的血清IL-13、IFN-γ的质量浓度。结果急性期毛细支气管炎患儿血清IL-13的质量浓度[(6.88±2.12)ng/L]明显高于对照组[(5.48±1.28)ng/L,P<0.01]。急性期毛细支气管炎患儿IFN-γ质量浓度[(10.71±2.44)ng/L]明显高于对照组[(9.20±1.54)ng/L,P<0.05];其中轻症组明显高于重症组和对照组(P<0.05),而重症组与对照组差异则无显著性(P>0.05)。结论急性期毛细支气管炎患儿血清IL-13质量浓度明显增高,提示IL-13参与了毛细支气管炎的发病过程,但所测质量浓度不能反映病情严重程度;IFN-γ在轻症毛细支气管炎组质量浓度明显增高,而重症组不增高,这可能与急性重症毛细支气管炎患儿IFN-γ产生受抑制有关。     

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.