地塞米松对儿童原发性系膜增生性肾炎外周血淋巴细胞CTLA-4表达和凋亡的影响

目的：原发性系膜增生性肾炎(MsPGN)的发病机制和糖皮质激素(GC)耐药机制虽然部分已明确,但其确切机制仍未阐明。细胞毒性T淋巴细胞相关抗原(CTLA-4)是T细胞活化的重要抑制性分子。本研究探讨MsPGN患儿外周血淋巴细胞CTLA-4表达和淋巴细胞凋亡及地塞米松(Dex)对其影响。方法：分别采用直接免疫荧光、流式细胞仪及Annexin V-FITC/PI双染流式细胞仪检测Dex处理或非Dex处理的MsPGN组和对照组体外培养外周血淋巴细胞膜CTLA-4的表达和其凋亡。结果：体外培养MsPGN组淋巴细胞CTLA-4的自然表达和Dex诱导的表达均较对照组低（P＜0.05）;MsPGN组淋巴细胞自然凋亡和Dex诱导的凋亡亦较对照组低（P＜0.05）,且两者均呈正相关性（P＜0.05）。结论：CTLA-4表达异常可能参与了MsPGN的发病机制及GC耐药机制。［中国当代儿科杂志,2009,11（12）：957-960］     

原发性肾病综合征与细胞毒性T淋巴细胞相关抗原-4基因启动子区-318位点基因多态性的相关性

目的探讨细胞毒性T淋巴细胞相关抗原-4(CTLA-4)与糖皮质激素(GC)耐药型原发性肾病综合征(PNS)中系膜增生性肾小球肾炎(MsPGN)的相关性。方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测36例PNS-MsPGN患儿和30例正常对照儿童CTLA-4基因启动子区-318位点基因型。结果肾病组CTLA-4基因启动子区-318位点基因型频率分别为CC型38.9%、TC型61.1%和TT型0,等位基因频率为C等位基因69.4%、T等位基因30.6%。肾病组各基因型及等位基因频率与对照组相比均无显著差异(P均>0.03)。结论CTLA-4基因启动子区-318位点基因C/T双态性同GC耐药型PNS-MsPGN患儿无相关性,提示该基点基因多态性可能不参与GC耐药型PNS-MsPGN的发病机制及耐药机制。     

过敏性紫癜患儿治疗前后淋巴细胞凋亡情况分析

目的 探讨细胞凋亡在过敏性紫癜（HSP）发病及病理过程中的作用。方法 对73例HSP患儿治疗前后采用原位末端标记技术进行制片，在光学显微镜下观察，计算出凋亡淋巴细胞的比率。结果 HSP急性期（治疗前）组淋巴细胞凋亡较正常对照组延迟（P〈0．01）；而治疗后组淋巴细胞凋亡较治疗前组明显增多（P〈0．01）。结论 过敏性紫癜（HSP）存在淋巴细胞凋亡延迟，经治疗随着临床症状好转，淋巴细胞凋亡逐渐恢复正常，推测可用促细胞凋亡的药物治疗HSP。     

肾小球肾炎患者外周血淋巴细胞Bcl-2和Fas的表达

评价外周血淋巴细胞凋亡在肾小球肾炎（GN）发病中的作用。方法用免疫组化、荧光显微镜及电子显微镜对GN进行病理分型,采用流式细胞术测定不同病理类型GN患者外周血单个核细胞（PBMC）Bcl－2、Fas表达水平。结果各型GNPBMCBcL－2表达水平均高于对照组（P＜0．01）,Fas均低于对照组（P＜0．01）;增殖型GN与非增殖型GN及增殖型GN中IgAN、MPGN、HSPN新月体组与非月体组间,Bcl－2、Fas表达水平存在显著差异,前者Bcl-2高于后者（P＜0．01）,Fas低于后者（P＜0．01）。结论Bcl－2过量表达,Fas系统功能不良,活化PBMC增多,导致肾小球肾炎。Bcl－2、Fas表达水平与GN病理类型及临床预后有一定联系。     

T 淋巴细胞毒相关抗原-４基因49位点多态性与儿童1型糖尿病相关性的Meta分析

目的：评价T 淋巴细胞毒相关抗原-4（CTLA-4）基因49位点多态性与儿童1型糖尿病（T1DM）的相关性。方法：检索PubMed、EBSCO、中国学术期刊全文数据库（CNKI）、中国生物医学文献数据库（CBM）、万方数据库,收集CTLA-4基因49位点多态性与儿童T1DM相关性的文献。用Meta分析的方法检测CTLA-4基因49位点的基因型和等位基因在儿童T1DM组与对照组中是否有差异。结果：共纳入10篇文献,共有T1DM 1084例,对照组1338例。根据各项研究的异质性检验结果,均利用固定效应模型对CTLA-4基因49位点的AG、GG、GG+AG基因型和G等位基因与T1DM相关性进行Meta分析,各项研究的合并OR值（95% CI）分别为1.13（0.97～1.33）、1.42（1.16～1.75）、1.20（1.03～1.40）、1.21（1.09～1.33）,提示T1DM组CTLA-4基因49位点G等位基因和GG、GG+AG基因型表达与对照组比较均有显著差异。结论：CTLA-4基因49位点G等位基因和GG、GG+AG基因型均与儿童T1DM的发生相关。     

淋巴细胞间质性肺炎

淋巴细胞间质性肺炎目前是一个病理学诊断，占围生期获得性免疫缺陷病毒感染婴儿的30－50％，临床表现为慢性间质性肺炎，须经肺部活检或支气管肺泡灌洗液检查肯定诊断，发病机制涉及免疫系统的非特异性刺激，人类免疫缺陷病毒（HIV）特异性刺激或EB病毒和HIV之间的协同作用，本文对有关淋巴细胞间质性肺炎的定义，病理和X线征象，发病机制，诊断技术新进展，临床表现和治疗等方面进行综述。     

急性淋巴细胞白血病糖皮质激素耐药的分子机制及其相关问题

糖皮质激素(GC)能特异性地诱导淋巴细胞周期停滞和凋亡,因此几十年来一直是治疗淋巴系统肿瘤,尤其是急性淋巴细胞白血病(ALL)的主要组成药物。然而对GC产生耐药性是临床ALL治疗中常见的难题,也是导致治疗失败的主要原因。GC耐药的确切的分子机制尚未阐明,与不同的疾病类型、治疗方案,尤其是患者的遗传背景有关。最近发现mTOR激酶介导的信号通路的活化能够诱导淋巴细胞对GC耐药,以及mTOR激酶抑制剂雷帕霉素能够有效地逆转ALL细胞对GC耐药作用提示联合应用mTOR激酶抑制剂和GC有望成为治疗ALL的有效措施。GC敏感试验是儿童ALL的一项主要预后判断指标,然而本身还有很多局限性,因此有必要寻找精确的特异性高的体外GC敏感试验。     

过敏性紫癜患儿外周血免疫功能与淋巴细胞凋亡的变化

为探讨过敏性紫癜患儿外周血免疫功能与淋巴细胞凋亡的变化及其相互关系，选择过敏性紫癜患儿58例正常对照组28例，应用单克隆抗体间接免疫荧光法检测CD3^ ,CD4^ ,CD8^ ,IgA,IgM,IgG采用免疫比浊法，外周血成熟淋巴细胞凋亡检测采用吖啶橙-溴化乙啶细胞内染色法，结果；过敏性紫癜患儿CD3^ ,CD4^ /CD8^ ,IgG明显降低，CD8^ ,IgM，淋巴细胞凋亡率（PCD）明显升高，与正常组比较有统计学意义，结果表明，过敏性紫癜患儿免疫功能降低并存在功能紊乱，外周血淋巴细胞凋亡率升高是导致以上变化的重要原因之一。     

尘螨变应性哮喘患儿外周血T淋巴细胞中协同刺激分子和细胞因子的异常表达及其意义

目的分析变应原刺激前后变应性哮喘患儿外周血T淋巴细胞表面协同刺激分子及胞内细胞因子表达的变化,探讨CD28家族不同协同刺激信号在变应性哮喘免疫病理机制中的作用。方法选取尘螨变应性哮喘患儿(哮喘组)和健康儿童(健康对照组)各30例,密度梯度离心法分离其外周血单个核细胞,应用免疫荧光标记和流式细胞术检测尘螨刺激前后体外培养的CD4+T淋巴细胞表面协同刺激分子CD28、可诱导共刺激分子(ICOS)和细胞毒T淋巴细胞相关抗原4(CTLA-4)的表达,运用细胞内染色技术检测CD4+T淋巴细胞内细胞因子γ干扰素(IFN-γ)、IL-4和IL-13的表达。并运用统计学方法比较哮喘组和健康对照组之间的差异。结果哮喘组患儿外周血CD4+T淋巴细胞表面CD28和ICOS的表达与健康对照组比较差异均无统计学意义(Pa>0.05),而CTLA-4的表达显著降低(P<0.01);细胞内IFN-γ表达水平显著升高(P<0.000 1),而IL-4和IL-13表达水平无明显变化(Pa>0.05)。经尘螨刺激后,体外培养的哮喘患儿外周血CD4+T淋巴细胞表面ICOS的表达较健康对照组儿童显著上调(P<0.000 1),CD28和CTLA-4的表达则无明显变化(Pa>0.05);细胞内细胞因子IL-4和IL-13的表达显著上调(Pa<0.000 1),而IFN-γ的表达则无明显差异(P>0.05)。结论变应性哮喘患儿外周血存在组成性的CTLA-4的表达下调和细胞因子IFN-γ的表达上调,介导Th1型细胞的异常活化;而变应原尘螨的刺激又介导了ICOS依赖的Th2型细胞的分化,导致Th1/Th2失衡。     

心肌炎患儿淋巴细胞凋亡的改变

目的 了解心肌炎患儿的淋巴细胞凋亡的改变,进一步探讨心肌炎的发病机理。方法 荧光法检测41例心肌炎患儿外周血淋巴细胞凋亡的改变,同时测定T淋巴细胞亚群与免疫球蛋白的变化。结果 心肌炎组淋巴细胞凋亡率高于正常组,且T淋巴细胞亚群CD4+,CD4+／CD8+明显下降,CD8+增高(P<0.01)。结论 淋巴细胞凋亡过度导致淋巴细胞总量失衡,T淋巴细胞及体液免疫功能均低下,可能是病毒或支原体感染后引起心肌炎的重要原因之一。     

小阴茎、小睾丸的诊断与治疗

小阴茎、小睾丸的病因复杂,临床诊断较为困难。下丘脑-垂体-性腺轴及雄激素合成和转化过程中任何一个环节出现异常,皆会影响阴茎和睾丸的发育。治疗应在明确诊断的基础上予以个体化方案。提倡重视小青春期内的早期诊断,以期早期治疗,使病变的危害降至最低。     

Age-dependency of alpha- and beta-adrenoceptors on thrombocytes and lymphocytes of asthmatic and nonasthmatic children

Among the possible mechanisms which may cause wheezing or asthmatic episodes a genetically determined -adrenoceptor blockade and a hyperresponsiveness of -andrenoceptors has been postulated. Evidence to support this hypothesis stems from an increased bronchial sensitivity to -blockers, a reduced formation of cyclic AMP in response to -adrenergic stimulation and enhanced -adrenergic responses in asthmatic subjects. The recent development of techniques for measuring the specific, high-affinity binding of radiolabeled -and -adrenergic antagonists made it possible to study - and -adrenoceptors in vitro. Based upon the assumption that a change in the number and/or affinity of adrenergic receptors might be a general phenomenon, we have performed - and -receptor binding studies on lymphocytes and platelets from wheezing infants and asthmatic children as well as of infants, children, and adults not suffering from these diseases.Using ¹²⁵[I]-cyanopindolol (ICYP) and ³[H]-yohimbine (HYOH) as highly specific ligands for - and -adrenoceptors, the following results were obtained: (1) Lymphocytes and platelets from control subjects and asthamatics bound similar amounts of ICYP and HYOH and thus showed no differences either in the number or the affinity of - and -adrenoceptors. Lymphocytes and platelets of wheezing and nonwheezing infants also bound the same amounts of the radioligands. (2) In asthmatic children receiving 4×2 puffs salbutamol -adrenoceptor were down-regulated and this may mimic -adrenoceptor blockade. (3) When subjects were divided into four categories according to age (0–5, 5–10, 10–20 years, adults) the number of -adrenoceptor binding sites showed an age-dependent increase. The number and affinity of -adreneceptor binding sites on platelets was neither influenced by age nor disease.It is concluded that the - and -adrenoceptors of wheezing infants and asthmatic children at least on blood cells are normal. However the -adrenoceptors show an age-dependent maturation process, which may account for an unresponsiveness to -adrenoceptor agonists in wheezing infants.Supported by a grant from the Ministerium für Wissenschaft und Forschung, NRWPresented at the 19th Workshop for Pediatric Research, University of Göttingen, March 10–11, 1983     

Specifics of histopathological and genetical diagnosis and classification of lymphomas in children and adolescents

Malignant lymphoma along with leukemias account for nearly half of all malignancies arising in childhood and adolescence. The correct tissue-based histopathological diagnosis of lymphomas results from a close interdisciplinary exchange between pediatric oncologists and hematopathologists. We describe here relevant features of lymphoma subtypes arising in the young age group, Burkitt lymphoma, precursor/lymphoblastic lymphomas, anaplastic large cell lymphoma and diffuse large B-cell lymphoma as well as primary mediastinal B-cell lymphoma and the rare pediatric follicular lymphomas. Special focus is put on specific diagnostic difficulties as well as new insights into biological features of pediatric lymphomas in comparison with their adult counterpart. In addition the relevance of newly defined lymphoma entities of the WHO-classification 2008, e.g. greyzone lymphomas, will be discussed for the young age group.     

Diagnosis and management of disorders of IGF-I synthesis and action

After a proper medical history, growth analysis and physical examination of a short child, followed by radiological and laboratory screening, the clinician may decide to perform genetic testing. We recently proposed several clinical algorithms that can be used to establish the diagnosis. GH insensitivity (primary IGF-I deficiency) can be caused by genetic defects in GHR, STAT5B, IGF1, IGFALS, which all have their specific clinical and biochemical characteristics. IGF-I resistance is seen in heterozygous defects of IGF1R. If besides short stature additional abnormalities are present, these should be matched with known dysmorphic syndromes. If no obvious candidate gene can be determined, a whole genome approach can be taken to check for deletions, duplications and/or uniparental disomies (SNP-array) or whole exome sequencing. Children with GHR defects, and presumably STAT5B and homozygous IGF1 defects, can be treated with rhlGF-I. Children with IGF1R defects and mild or heterozygous IGF1 defects respond to GH treatment.