Toll样受体9及其配体研究进展

哺乳动物的免疫系统通过模式识别受体来识别病原微生物,Toll样受体(TLR)9是主要的先天性免疫模式识别受体之一。TLR9的天然配体是病毒和细菌基因组中的非甲基化胞嘧啶-磷酸-鸟嘌呤二核苷酸序列(CpG),实验研究中多采用人工合成的具有抗核酶降解作用的类似物——含有CpG基序的寡聚脱氧核苷酸。TLR9被其配体激动后可使机体产生强烈的Th1型优势免疫应答,在变态反应性疾病、感染性疾病和癌症方面的治疗中有广泛的应用前景。     

慢性鼻窦炎儿童的免疫状况分析

目的:分析慢性鼻窦炎患儿细胞免疫和体液免疫功能状况及其临床意义.方法:对43例3～5岁的慢性鼻窦炎患儿(病例组)和46例健康对照儿童(对照组)进行淋巴细胞亚群及免疫球蛋白测定.淋巴细胞亚群采用美国贝克曼库尔特EPICS-XL流式细胞仪进行检测,血清免疫球蛋白采用免疫比浊法检测,唾液sIgA测定用单向免疫扩散法.结果:与对照组比较,病例组淋巴细胞总数、CD3+、CD4+、CD8+和CD16+CD56+细胞数量均少于对照组,差异有统计学意义(t值9.258～50.948,均P＜0.01),两组CD19+细胞数量差异无统计学意义.病例组CD4+%、CD16+CD56+%和CD4/CD8少于对照组,差异有统计学意义(t值4.068～4.700,均P＜0,01);病例组CD19+%多于对照组,差异有统计学意义(t=9.514.P＜0.01).两组CD3+%和CD8+%比较差异无统计学意义.病例组血清IgG和唾液sIgA低于对照组,差异有统计学意义(t值为3.648和2.569,均P＜0.05).两组血清IgA和血清IgM水平差异无统计学意义.结论:慢性鼻窦炎患儿淋巴细胞总数、CD3+、CD4+、CD8+和CD16+ CD56+细胞数量降低,血清IgG和唾液sIgA水平也降低.慢性鼻窦炎患儿存在细胞免疫和体液免疫功能低下.     

孟鲁司特联合匹多莫德治疗儿童支气管哮喘35例的疗效观察

目的 探讨孟鲁司特联合匹多莫德口服治疗间歇发作支气管哮喘患儿的临床疗效及安全性.方法 对69例支气管哮喘患儿采用随机数字表分为两组:治疗组35例用孟鲁司特钠咀嚼片口服12个月,前3个月同时服用匹多莫德颗粒;对照组34例仅给予孟鲁司特钠咀嚼片口服12个月.两组均随访12个月,记录两组患儿临床表现情况、血液淋巴细胞亚群、免疫球蛋白及不良反应.结果 与对照组比较,治疗组的哮喘发作次数、上呼吸道感染次数和下呼吸道感染次数均少于对照组,差异均有统计学意义.治疗组在治疗后3个月时CD3、CD4、CD8,CD4/CD8比值及血清IgA升高,与治疗前比较有统计学意义;对照组与治疗前比较差异无统计学意义.两组不良反应发生率差异无统计学意义.结论 联合使用孟鲁司特和匹多莫德口服可显著减少哮喘发作次数,疗效明显优于单独使用孟鲁司特组,两种药物联合使用无明显不良反应.     

新生儿高胆红素血症巨细胞病毒感染状况分析

随着人们对新生儿黄疸的重视,因黄疸就诊、住院的新生儿逐年增多。人类巨细胞病毒（Cytomegalo virus,CMV）感染是新生儿黄疸的重要原因,可导致黄疸消退延迟或复现、肝脾肿大、肝功能异常等。本文对216例新生儿高胆红素血症患儿进行CMV感染情况分析和随访,以期了解新生儿高胆红素血症患儿中CMV感染情况。     

Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

目的 研究Toll样受体9(toll-like receptor 9,TLR9)基因启动子区单核苷酸多态性在浙江汉族儿童中的分布,探讨其与哮喘易感性及其表型之间的相关性.方法 对312例变应性哮喘患儿(哮喘组)和339名健康儿童(对照组)采用DNA直接测序法检测TLR9基因-1486(rs187084)和-1237(rs5743836)单核苷酸多态性;采用ELISA法检测两组不同基因型血清干扰素γ(interferon-γ,IFN-γ)、白细胞介素12(interleukin-12,IL-12)和白细胞介素4(interleukin-4,IL-4)水平;采用化学发光法检测血清总免疫球蛋白E(immunoglobulin E,IgE)水平;采用酶免疫荧光法检测血清变应原特异IgE.结果 (1)哮喘组和对照组均存在-1486位点T→C突变,哮喘组TT、TC和CC 3种基因型的频率分别是3 8.8%、48.4%和12.8%,对照组分别是41.0%、44.3%和14.7%;未发现-1237位点存在多态性.(2)哮喘组和对照组-1486位点各基因型的频率分布差异无统计学意义(P＞0.05),年龄分层后比较差异也无统计学意义(P＞0.05).(3)哮喘组-1486位点3种基因型的血清IFN-γ和IL-4水平差异有统计学意义(P＜0.01),CC基因型的IFN-γ水平较低而IL-4水平较高;对照组2种细胞因子的差异无统计学意义(P＞0.05).哮喘组和对照组血清IL-12水平在3种基因型间差异均无统计学意义(P＞0.05).(4)哮喘组-1486位点不同基因型血清总IgE水平差异无统计学意义(P＞0.05).结论 浙江汉族儿童不存在TLR9基因-1237位点多态性.TLR9基因-1486 C/T位点单核苷酸多态性与浙江汉族儿童哮喘易感性、血清IL-12及总IgE水平无关;-1486 C/T位点多态性与哮喘患儿血清IFN-γ和IL-4水平有关联,CC基因型的IFN-γ水平较低而IL-4水平较高.

Abstract：

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

氟替卡松联合细菌溶解产物对儿童哮喘的疗效观察

目的探讨丙酸氟替卡松吸入气雾剂联合细菌溶解产物（泛福舒）对间歇发作的支气管哮喘儿童的临床疗效和安全性。方法2009年2～6月收治的支气管哮喘患儿93例随机分为两组：实验组47例在吸入丙酸氟替卡松的基础上加用细菌溶解产物胶囊,口服3个月;对照组46例仅给予丙酸氟替卡松吸入治疗。两组均随访12个月,记录两组患儿临床表现、血清细胞因子、免疫球蛋白及不良反应。结果与对照组比较,治疗后实验组的上呼吸道感染次数（t=10．731）、下呼吸道感染次数（Z=3．548）、哮喘发作次数（t=4．758）、每次发作的咳嗽天数（Z=3．267）、发热天数（t=-2．401）和抗生素使用天数（t=-2．079）均少于对照组,差异均有统计学意义（均P〈0．05）。与对照组比较,实验组在治疗后3个月时血清IFN-γ（χ2=-10．417）、血清IgA（t=5．928）和唾液sIgA（t=-13．627）高于对照组,血清IL-4（χ2=-6．106）低于对照组,差异有统计学意义（均P〈0．05）。两组不良反应发生率差异无统计学意义（χ2=2.60t,P〉0．05）。结论间歇发作的支气管哮喘患儿在使用丙酸氟替卡松吸人的基础上加用细菌溶解产物口服,可显著减少哮喘发作次数,疗效明显优于单独使用丙酸氟替卡松组,两药联合使用无明显不良反应。     

孟鲁司特和卡介苗多糖核酸对哮喘大鼠血单个核细胞中STAT5b mRNA和IL-4 mRNA表达的影响

目的：研究孟鲁司特和卡介苗多糖核酸对哮喘大鼠血单个核细胞中信号转导子和转录激活子5b(STAT5b) mRNA和白细胞介素4（IL-4）mRNA的转录表达的影响。方法：52只体重为140~200 g清洁级Sprague-Dawley雄性大鼠,随机分为4组：哮喘组、孟鲁司特组、卡介苗多糖核酸组和正常对照组。用卵白蛋白制备大鼠哮喘模型。计数血中嗜酸性粒细胞(EOS);应用双抗体夹心酶联免疫吸附试验测定血浆中IL-4和γ-干扰素（IFN-γ）浓度;采用SYBR GREEN I荧光实时定量PCR法测定STAT5b mRNA和IL-4 mRNA的相对表达量。结果：①哮喘组血单个核细胞中STAT5b mRNA和IL-4 mRNA的表达量高于其他各组（P<0.01）;在孟鲁司特组和卡介苗多糖核酸组中两者的表达量相近（P>0.05）。哮喘组除了IFN-γ水平低外,其余无论是STAT5b mRNA,IL-4 mRNA,EOS绝对值等都是4组中最高的（P<0.01）;除哮喘组外,其余组的各项指标相近（P>0.05）。②STAT5b mRNA表达量与IL-4 mRNA表达量、IL-4浓度、EOS绝对值呈正相关(P<0.01),与IFN-γ浓度呈负相关（P<0.01）。结论：哮喘大鼠血单个核细胞中STAT5b mRNA和IL-4 mRNA的表达增强且呈正相关;孟鲁司特和卡介苗多糖核酸可以下调STAT5b mRNA和IL-4 mRNA的表达,可能为其抑制哮喘气道炎症形成的重要机制之一。［中国当代儿科杂志,2009,11（2）：133-137］     

细菌溶解产物联合孟鲁司特治疗儿童哮喘的疗效观察

目的：探讨细菌溶解产物（泛福舒）联合孟鲁司特口服对间歇发作的支气管哮喘患儿的临床疗效及安全性。方法：173例间歇发作的支气管哮喘患儿随机分为两组：实验组87例,用孟鲁司特咀嚼片口服12个月,前3个月同时服用细菌溶解产物;对照组86例,仅给予孟鲁司特咀嚼片口服12个月。两组均随访12个月,记录两组患儿临床表现、血液淋巴细胞亚群、免疫球蛋白及不良反应情况。结果：173例中有156例（实验组76例,对照组80例）完成随访12个月。与对照组比较,实验组的哮喘发作次数、呼吸道感染次数、发热天数、咳嗽天数与抗生素使用天数均少于对照组,差异均有统计学意义（P〈0.05）。治疗后3个月时,实验组CD3^＋%、CD4^＋%、CD4^＋/CD8^＋比值、血清IgA和唾液sIgA高于对照组,差异均有统计学意义（P〈0.01）;两组CD8^＋%、IgG、IgM和IgE差异无统计学意义（P〉0.05）。两组不良反应发生率比较差异无统计学意义（校正χ^2=0.856,P〉0.05）。结论：间歇发作的支气管哮喘患儿联合使用细菌溶解产物和孟鲁司特口服可显著减少哮喘发作次数,疗效明显优于单独使用孟鲁司特,两种药物联合使用无明显不良反应。