IL-13基因多态性与RSV感染后婴幼儿喘息及哮喘关系的研究

目的 探讨IL-13基因Arg110Gln多态性在RSV感染后婴幼儿喘息发生和哮喘发病中的作用.方法 哮喘患儿和止常儿童各96例,RSV感染后喘息婴幼儿112例,取其颊黏膜细胞,提取DNA,用实时PCR法对IL-13 Arg110Gln位点进行基因分型.结果 IL-13 Arg110Gln位点的基因型和等位基因频率分布在哮喘组和对照组间差异有高度统计学意义(P<0.01);哮喘组的A等位基因和A/A纯合子基因型频率明显高于对照组(分别为68.2%和47.4%,46.9%和29.2%,P<0.01).IL-13 Arg110Gln位点的基因型和等位基因频率分布在哮喘组和RSV感染后喘息组间差异无统计学意义(P>0.05).结论 IL-13 Arg110Gln位点G→A变异与RSV感染后婴幼儿喘息发生和哮喘发病相关,两者在该位点上可能存在相似的遗传基础.     

IL-13 基因+2044G/A 多态性与儿童哮喘风险和血清IL-13 水平的相关性研究

目的探讨IL-13基因+2044G/A多态性与儿童哮喘发病风险及哮喘儿童血清IL-13水平的关系。方法采用PCR-RFLP和ELISA方法,对90例支气管哮喘患儿(哮喘组)和82例健康儿童(对照组)IL-13基因+2044G/A多态性和血清IL-13水平进行检测,计算基因型、等位基因频率及其所对应的血清IL-13水平。结果哮喘组和对照组IL-13+2044G/A多态性3种基因型的分布差异有统计学意义(P0.001);与GG基因型比较,GA基因型儿童哮喘风险增加3.52倍(95%CI:1.82~6.82),AA基因哮喘风险增加3.71倍(95%CI:1.06~12.97)。哮喘组A等位基因频率为36.1%,高于对照组的18.3%,差异有统计学意义(P0.001)。与G等位基因比较,A等位基因携带儿童哮喘风险增加2.53倍(95%CI:1.53~4.16)。哮喘组患儿血清IL-13水平为151.82(134.33~166.68)ng/L,对照组为79.00(58.74~111.09)ng/L,两组差异有统计学意义(P0.001)。哮喘组GG、GA和AA基因型的血清IL-13水平均分别高于对照组,差异有统计学意义(P均0.01)。哮喘组GG、GA和AA 3种基因型之间血清IL-13水平的差异也有统计学意义(P0.01),其中GA和AA基因型血清IL-13水平高于GG基因型。结论 IL-13基因+2044G/A多态性与支气管哮喘儿童血清IL-13水平具有关联性,且A等位基因可能造成了哮喘儿童血清高IL-13水平,携带A等位基因的儿童哮喘风险增加。     

TNF-α-308G/A和IL-6-174G/C基因多态性与呼吸道合胞病毒毛细支气管炎的相关性

目的：探讨肿瘤坏死因子(TNF)-α-308G/A、白细胞介素(IL)-6-174G/C基因多态性与呼吸道合胞病毒((RSV) 毛细支气管炎易感性的关系。方法：应用聚合酶链反应 限制性酶切片段长度多态性技术（PCR-RFLP）检测150例RSV毛细支气管炎患儿(病例组)和120例正常对照儿童(对照组)TNF-α-308G/A和IL-6-174G/C的基因多态性。结果：①病例组TNF-α-308位点GG、GA、AA基因型频率分别为68.0%、28.0%、4.0%,G、A等位基因型频率分别为82.0%,18.0%;对照组TNF-α-308位点GG,GA,AA基因型频率分别为80.8%,19.2%,0,G、A等位基因型频率分别为90.4%,9.6%;两组基因型及等位基因频率差异有显著性意义,病例组A等位基因的频率明显高于对照组(χ2=5.665,7.726,均P＜0.05);与G等位基因携带者相比,A等位基因携带者患RSV毛细支气管炎的风险增加了2.071倍(OR=2.071, P＜0.05);②病例组和对照组IL-6-174G/C位点均只见GG基因型。结论：TNF-α-308A等位基因与RSV毛细支气管炎的易感性相关,其可能是影响RSV毛细支气管炎发病的一个重要候选基因;温州地区汉族儿童未发现IL-6-174G/C基因多态性。［中国当代儿科杂志,2009,11（10）：821-824］     

IL-12 p40 3''''非翻译区rs3212227单核苷酸多态性与呼吸道合胞病毒感染后喘息发生的关系研究

目的 探讨IL-12 p40基因3'非翻译区(3'URT)单核苷酸多态性位点(rs3212227)与呼吸道合胞病毒(RSV)感染后病情严重度的关系.方法 对2004-2006年儿内呼吸科收治的呼吸道感染患儿用直接免疫荧光法进行鼻咽部分泌物呼吸道病原抗原检测,选取RSV阳性患儿共175例,按临床表现分为喘息组(112例)和非喘息组(63例),用Taqman探针检测法对两组患儿行IL-12 p40rs3212227位点单核苷酸多态性分析.两组患儿血清总IgE定量采用Unicap体外检测系统测定.结果 RSV感染后喘息组rs3212227位点等位基因A和C的发生频率分别为48.2%、51.8%,非喘息组分别为29.4%、70.6%,两组等位基因频率差异有统计学意义.喘息组A/A纯合子、C/C纯合子和A/C杂合子基因型频率分别为21.4%、25.0%和53.6%,非喘息组A/A纯合子、C/C纯合子和A/C杂合子基因型频率分别为9.5%、50.8%和39.7%,两组基因型频率差异有统计学意义.尽管喘息组患儿血清总IgE水平显著高于非喘息组患儿(P＜0.05),但喘息组rs3212227位点A/A纯合子、C/C纯合子和A/C杂合子患儿之间的血清总IgE水平差异无统计学意义(P＞0.05).结论 IL-12 p40rs3212227位点单核苷酸多态性与RSV感染后喘息的发生有关,但与血清总IgE水平无关.     

Clara细胞分泌蛋白10基因G38A多态性与5岁以下喘息患儿的相关性

目的探讨5岁以下喘息患儿Clara细胞分泌蛋白10(CC10)基因G38A位点多态性与喘息发病机制的关系。方法随机选取于本院就诊的5岁以下反复喘息患儿120例,分为有特应质高危因素的喘息Ⅰ组(n=67)(湿疹45例,父母或父母一方有哮喘病史13例,变应性鼻炎5例,变应性皮炎4例)和无特应质高危因素的喘息Ⅱ组(n=53);对照组为本院外科近期无感染疾病史、择期进行手术的术前患儿(n=55)。采用PCR-限制性片段长度多态性分析对喘息组和对照组患儿CC10 G38A位点基因型频率和等位基因频率进行检测,比较3组间CC10 G38A位点基因型频率和等位基因频率。结果喘息Ⅰ组、喘息Ⅱ组和对照组3种基因型AA、GA、GG分布频率分别为20.9%、44.8%、34.3%,9.4%、32.1%、58.5%、9.1%、31.0%、60.0%;喘息Ⅰ组和喘息Ⅱ组CC10基因G38A位点基因型频率比较差异有统计学意义(P<0.05);喘息Ⅰ组和对照组CC10基因G38A位点基因型频率比较差异亦具有统计学意义(P<0.05)。喘息Ⅰ组、喘息Ⅱ组和对照组38A和38G等位基因频率分别为43.3%、56.7%,25.5%、74.5%,24.5%、75.5%;喘息Ⅰ组和喘息Ⅱ组、喘息Ⅰ组和对照组比较差异均有统计学意义(Pa<0.05)。结论喘息患儿与哮喘存在相同的基因分布频率,发生哮喘的危险性高;对于CC10基因具有A等位基因的喘息患儿应密切关注。     

呼吸道合胞病毒毛细支气管炎与白细胞介素-10-592A/C基因多态性的相关性研究

目的 探讨白细胞介素-10-592A/C基因多态性与呼吸道合胞病毒(RSV)毛细支气管炎易感性、病情的关联和对血清白细胞介素(IL)-10的影响.方法 采用聚合酶链反应-限制性片段长度多态性检测100例汉族RSV毛细支气管炎患儿(病例组)和100例健康儿童(对照组)IL-10-592A/C位点单核苷酸多态性;应用ELISA法检测病例组血清IL-10水平.结果 病例组IL-10-592A/C位点基因型频率分别为AA 44%、AC 38%、CC 18%,等位基因频率分别为A 63%、C 37%;对照组基因型频率分别为AA 41%、AC 42%、CC 17%,等位基因频率分别为A 64%、C 36%;两组基因型及等位基因频率比较差异均无显著性(χ2=0.33,P>0.05;χ2=0.43,P>0.05).病例组IL-10-592A/C位点不同基因型患儿血清IL-10水平比较差异无显著性(F=0.87,P>0.05).IL-10-592A/C位点基因型频率在轻度和中重度患儿之间的差异无显著性(χ2=2.67,P>0.05).结论 IL-10-592A/C位点基因多态性与RSV毛细支气管炎不存在关联.     

儿童外伤性脑梗死与蛋白S基因多态性的关系

目的 探讨儿童外伤性脑梗死发生发展与蛋白S基因多态性的关系.方法 应用聚合酶链反应(PCR)和限制性酶切片段长度多态性(RFLP)技术对61例外伤性脑梗死患儿蛋白S基因A2148G多态性进行分析.结果 蛋白S基因型与等位基因频率分别为:GG基因型57.38%、GA基因型37.70%、AA基因型6.55%,G等位基因76.23%、A等位基因23.77%;外伤性脑梗死患儿GG基因型和G等位基因频率明显高于对照组,差异有统计学意义(P ＜ 0.05);蛋白S GG基因型发生外伤性脑梗死的相对危险高,OR值为3.14.结论 蛋白S GG基因型可能是儿童外伤性脑梗死的易感基因型.开展儿童外伤性脑梗死与蛋白S基因多态性关系的研究对于探讨儿童外伤性脑梗死的发病机制,寻找儿童外伤性脑梗死的保护基因和易感基因及预防和及时治疗儿童外伤性脑梗死有重要意义.     

白细胞介素8-251位点基因多态性与呼吸道合胞病毒毛细支气管炎及毛细支气管炎后婴幼儿喘息的关系

目的探讨IL-8启动子-251A／T基因多态性与呼吸道合胞病毒（RSV）致毛细支气管炎（简称毛支）及毛支后婴幼儿喘息的相关性。方法应用聚合酶链反应．限制性片段长度多态性（PCR—RFLP）技术检测320例RSV毛支患者及272例正常对照者IL-8基因-251A／T多态性,酶联免疫吸附试验（ELISA）法检测血清IL-8、IgE浓度。并对人组的毛支患儿随访3年,记录婴幼儿毛支后喘息再发的情况。结果（1）RSV毛支组和对照组IL-8-251位A等位基因频率分别为45．6％、37．7％,两组问比较差异有统计学意义（P〈0．05）;（2）基因型TT、AT、AA的毛支患儿血清IL-8浓度分别为（17±6）ng／L、（21±7）ng／L、（24±9）ng／L,三种基因型间比较差异有统计学意义（P〈0．01）;（3）RSV毛支后喘息组和未再喘息组IL-8-251位A等位基因频率分别为54．6％、35．8％,两组间比较差异有统计学意义（P〈0．05）。结论IL-8启动子-251A／T基因多态性与RSV毛支易感性相关,且携带IL-8-251A等位基因的患儿在RSV毛支后更容易出现喘息。     

白细胞介素-10基因多态性与儿童肠道病毒71型感染的相关性研究

目的 探讨白细胞介素-10（IL-10）基因位点-1082A/G、-819C/T和-592C/A多态性与IL-10水平及儿童肠道病毒71型（EV71）感染程度的关系。方法 选取EV71感染的手足口病患儿137例为研究对象,其中轻症组91例,重症组46例,另选取行健康体检儿童122例为健康对照组,采集临床数据。采用酶联免疫吸附试验（ELISA）检测血清IL-10水平。采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析技术检测IL-10基因位点-1082A/G、-819C/T和-592C/A的多态性。结果 与健康对照组相比,EV71感染组患儿的-1082位点AA基因型频率和A等位基因频率更高（P < 0.05）。在EV71感染组中,重症组-1082位点AA基因型频率和A等位基因频率高于轻症组（P < 0.05）。两组间IL-10基因位点-819C/T和-592C/A多态性的分布比较差异均无统计学意义（P > 0.05）。重症组患儿血清IL-10水平明显高于轻症组和健康对照组（P < 0.05）。IL-10-1082 AA基因型、-819 TT基因型和-592 AA基因型与IL-10的低表达有关（P < 0.05）。在单倍型构建上,EV71感染组GCC单倍型的频率低于健康对照组（P < 0.05）。在EV71重症感染组中,ATA单倍型患儿的IL-10水平较其他单倍型显著降低,而GCC患儿的IL-10水平较其他单倍型显著升高（P < 0.05）;轻症组和健康对照组各单倍型间的IL-10水平比较差异无统计学意义（P > 0.05）。结论 IL-10基因多态性与IL-10表达水平及EV71感染严重程度有关。     

哮喘儿童白细胞介素13基因型与表型的关联研究

目的探讨哮喘儿童白细胞介素13(IL-13)Arg130Gln基因型与血清总IgE、IL-13水平及过敏原表型之间的关系。方法采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)法,对130例哮喘患儿和100例健康对照儿童进行IL-13Arg130Gln基因多态性的检测,并用酶免法检测血清总IgE、IL-13水平和空气过敏原。结果基因型分布及等位基因频率哮喘组与对照组比较差异均无显著性,哮喘组突变型个体的Log10IL-13水平高于未突变型个体的水平(P<0.05),哮喘组突变纯合型个体的Log10IgE水平高于突变杂合型及未突变型个体的水平(P<0.05),哮喘组及对照组突变型与未突变型的过敏原阳性率比较差异均无显著性。结论IL-13Arg130Gln突变不是中国长春地区儿童哮喘的易感基因,但此突变可能与血清IL-13蛋白质水平升高有关,突变纯合型可能与血清总IgE水平相关联。     

Clara cell protein 10 polymorphism is not associated with severe respiratory syncytial virus infection*

Aim: To investigate a possible correlation between severe respiratory syncytial virus (RSV) infection and single‐nucleotide polymorphism (SNP) in the Clara cell protein 10 (CC10) gene (A38G). Methods: Children hospitalized with severe RSV lower respiratory tract (LRTI) infection at Karolinska University Hospital, Stockholm, during three subsequent RSV seasons were selected and genotyped. Age‐matched controls were used. The two groups were compared regarding SNP in the CC10 gene (A38G) and regarding later development of wheezing. Results: No correlation was found between the genotype CC10 +38AA and severe RSV infection or wheezing during follow‐up. Wheezing (p = 0.022), frequent wheezing (≥3 episodes) (p = 0.042) as well as ever wheezing (p = 0.0022) occurred significantly more often after discharge in children hospitalized for RSV compared with the control group. Conclusion: Single‐nucleotide polymorphism in the CC10 gene (A38G) does not seem to be involved in the severity of RSV infection or wheezing. In agreement with other studies, we found that children with severe RSV are at increased risk of future wheezing. The latter finding implies that the studied population represented children with severe LRTI, which supports that a correlation between the genotype CC10 +38AA and severe RSV is less likely.     

Nasopharyngeal aspirate cytokine levels 1 yr after severe respiratory syncytial virus infection

Respiratory syncytial virus (RSV) infection is an important cause of recurrent wheezing in infants. Nevertheless, the link between RSV infection and wheezing has yet to be elucidated at the molecular level. Here, we present a preliminary study on the evolution of the immune response in the respiratory tract at long‐term after RSV infection. Twenty‐seven immune mediators were profiled in nasopharyngeal aspirates (NPAs) obtained from 20 children hospitalized due to a severe infection by RSV at discharge from hospital and again 1 yr later. The same mediators were profiled in parallel in NPAs from 12 healthy controls. In the year following discharge, 85% (17/20) of children of the RSV group suffered at least one episode of wheezing documented by the pediatrician. On the contrary, wheezing episodes were observed only in 25% (3/12) of children in the control group. While most of the mediators profiled returned to normal levels by 1 yr after discharge from hospital, RSV children showed a persistent nasal hyper‐secretion of VEGF, G‐CSF, IL‐10, IL‐6, IFN‐γ, IL‐7 and IL‐13. In previous works VEGF, IL‐10 and IFN‐γ have been put in relation with the pathogenesis of post‐virus induced asthma. G‐CSF, IL‐6, IL‐7 and IL‐13 are increased in respiratory and plasma samples of asthmatic patients. Here, we evidence for the first time a persistent elevation of these mediators as late as 1 yr after severe RSV disease resolution, reinforcing their possible implication in the pathogenesis of wheezing.     

白细胞介素-4受体基因多态性及IgE水平与哮喘预测指数相关性研究

目的研究新疆维吾尔自治区儿童白细胞介素-4受体(IL-4R)基因多态性及Ig E水平与哮喘预测指数(API)的相关性。方法选取167例API(+)、187例API(-)及203例健康婴幼儿(对照组),应用PCR聚合酶链反应和DNA测序法进行基因分型。同时ELISA法检测三组婴幼儿血清Ig E水平。结果 IL-4R基因Arg551Gln位点各基因型频率分布在API(+)组、API(-)组、对照组三组中差异无统计学意义(P0.05);API(+)组血清Ig E水平明显高于API(-)及对照组(P0.01);API(+)患儿中2岁者血清Ig E水平明显低于≥2岁者(P0.01)。结论 Arg551Gln位点多态性与API结果无相关性,而API阳性与Ig E水平升高存在关联性,年龄≥2岁是API阳性的儿童Ig E水平升高的风险因素。     

喘息婴幼儿外周血气道炎症相关介质水平的研究

目的 通过检测喘息婴幼儿外周血相关炎症介质水平,从辅助性T细胞（Th）1/Th2失衡及气道炎症两个方面探讨婴幼儿喘息发生的可能机制。方法 选取急性喘息发作婴幼儿50例为喘息组,25例健康婴幼儿为健康对照组。喘息组患儿依据喘息发生次数分为首次喘息组（首发组）25例和反复喘息组（反复组,发作次数≥ 2次）25例;根据是否存在哮喘发生的高危因素分为有高危因素组22例和无高危因素组28例;根据病原学检测结果分为病原学阳性组23例和病原学阴性组27例。检测各组外周血Th1细胞因子白介素（IL）-2,Th2细胞因子IL-4、IL-5、IL-13、转化生长因子-β1（TGF-β1）,以及总IgE （TIgE）水平。喘息组患儿同时送检外周血嗜酸性粒细胞（EOS）计数,并采集标本进行呼吸道病原学检测。结果 喘息组外周血IL-4、IL-5、IL-13、TGF-β1及TIgE水平较健康对照组均明显升高（P < 0.05）;外周血IL-2、IL-4、IL-5、IL-13、TGF-β1及TIgE水平在首发组与反复组,有哮喘高危因素组与无哮喘高危因素组,以及病原学阳性组与病原学阴性组之间比较,差异均无统计学意义（P > 0.05）。相关性分析表明喘息组外周血EOS计数与IL-4水平呈正相关（P < 0.01）;IL-4水平与IL-5、IL-13水平呈正相关（P < 0.01）;IL-5水平与IL-13水平呈正相关（P < 0.01）;IL-2水平与TGF-β1水平呈正相关（P < 0.05）。结论 喘息婴幼儿存在Th1/Th2失衡,表现为Th2优势表达;IL-4、IL-5、IL-13、TGF-β1及IgE共同参与了婴幼儿喘息性疾病的发病过程。喘息婴幼儿存在气道炎症,与喘息发作次数、是否存在哮喘高危因素及病原学检测是否阳性无关。     

IL-13和TNF-α在肺炎支原体肺炎儿童血清中的表达及意义

目的：观察白细胞介素13（interleukin-13,IL-13）和肿瘤坏死因子α（tumor necrosis factor alpha,TNF-α）在小儿肺炎支原体肺炎中的变化。方法:酶联免疫方法检测40例肺炎支原体肺炎伴喘息患儿、40例肺炎支原体肺炎不伴喘息患儿、其他病原菌感染肺炎40例（普通肺炎组）及40例正常对照组儿童的血清IL-13和TNF-α的浓度。结果:肺炎支原体肺炎患儿血清IL-13、TNF-α的水平高于正常对照组及普通肺炎组;肺炎支原体肺炎伴喘息患者的IL-13、TNF-α的水平分别为214.6±67.2 ng/L,0.55±0.13 ng/L,高于不伴喘息者（189.6±52.1 ng/L, 0.42±0.16 ng/mL）,差异均有显著性（P<0.01）。结论：IL-13、TNF-α浓度增高可能在小儿肺炎支原体肺炎导致喘息样发作中起重要作用。［中国当代儿科杂志,2010,12（4）：275-277］     

Age and sex as factors of response to RSV infections among those with previous history of wheezing

Although enhanced immune reaction caused by the respiratory syncytial virus (RSV) in allergen-sensitized animal model has been reported, RSV illnesses in children already sensitized or having recurrent wheezing episodes have not been completely studied. In addition, the reason for male dominances in RSV infection at young ages was also inconclusive. Therefore, gender analysis in recurrent wheezing children with RSV infection can shed light on asthma pathogenesis. We studied the clinical features and the laboratory data of RSV infections in children who had recurrent wheezing histories. The subjects with RSV infection consisted of 98 boys and 58 girls. The children under 4 yr of age were 123 (78.8%) in number. Children with pneumonia were 78 and those with febrile episode were 119. Children above 1 yr of age were highly sensitized with mite antigen (75/96, 78.1%). The clinical symptoms and signs differed according to their ages. Children in each age group behaved differently in their immune reaction to RSV. Above all, 3-yr-old children deteriorated clinically during acute RSV infection, accompanied by transient elevated C-reactive protein (CRP) and suppressed blood eosinophil counts. Clinical features differed in several points between boys and girls. In general, the white blood cell count and the CRP levels were higher in girls in every age group. Blood eosinophil counts at the acute illness were significantly higher in boys than girls aged 2 and 3< yr. Age and gender comparison in already sensitized children might suggest a clue to asthma pathogenesis.