老年支气管哮喘患者血浆中IL-9检测及临床意义

目的 检测老年支气管哮喘患者血浆中IL-9的水平,探讨IL-9在支气管哮喘发生发展中的意义.方法 收集老年支气管哮喘患者和健康对照外周血标本.采用酶联免疫吸附试验(ELISA)检测血浆中IL-9和总IgE浓度.结果 老年支气管哮喘急性发作期患者血浆IL-9水平显著高于健康对照组(P<0.05),而临床缓解组与健康对照组差异无统计学显著性意义(P>0.05).老年支气管哮喘急性发作期患者血浆IL-9水平与总IgE浓度呈正相关.结论 老年支气管哮喘患者血浆中IL-9水平升高,与疾病的发生发展关系密切.     

支气管哮喘患儿血浆中IL-9的检测及临床意义

目的检测支气管哮喘患儿血浆中白细胞介素-9(IL-9)的水平,探讨IL-9在支气管哮喘发生发展中的意义。方法收集支气管哮喘患儿和健康对照外周血标本。采用酶联免疫吸附试验(ELISA)检测血浆中IL-9和总IgE浓度。结果支气管哮喘急性发作期患儿血浆IL-9水平高于健康对照组(P0.05),而临床缓解组和健康对照组比较,差异无统计学意义(P0.05)。支气管哮喘急性发作期患儿血浆IL-9水平与总IgE浓度呈正相关。结论支气管哮喘患儿血浆中IL-9水平升高,与疾病的发生发展关系密切。     

支气管哮喘患儿外周血Th17和Th9细胞及其细胞因子的表达及临床意义研究

目的检测儿童支气管哮喘患者T细胞17(Th17)、外周血辅助性T细胞9(Th9)的比例及血清中白细胞介素-17(IL-17)、白细胞介素-9(IL-9)和总IgE的表达,探讨其在儿童支气管哮喘发病中的作用及临床意义。方法选取46例哮喘患儿,分为哮喘发作组(26例)、哮喘缓解组(20例),并设健康对照组(20例)。采用流式细胞术分别检测3组外周血中Th17细胞和Th9细胞的比例;应用酶联免疫吸附试验(ELISA)检测3组儿童血清中IL-17和IL-9的水平;应用特定蛋白分析仪检测3组儿童血清中总IgE的水平。结果与哮喘缓解组、健康对照组相比,哮喘发作组外周血Th17和Th9细胞比例较高,差异有统计学意义(P0.05)。与健康对照组相比,哮喘缓解组外周血Th17和Th9细胞比例较高,比较差异有统计学意义(P0.05)。与哮喘缓解组、健康对照组相比,哮喘发作组血清中IL-17、IL-9和总IgE水平较高,差异有统计学意义(P0.05)。与健康对照组相比,哮喘缓解组血清中IL-17、IL-9和总IgE水平较高,差异有统计学意义(P0.05)。相关性分析结果表明,支气管哮喘患儿血清中IL-17、IL-9与总IgE水平均呈正相关(r值分别为0.717、0.491、0.786,P0.05)。结论支气管哮喘患儿外周血Th17和Th9细胞及其细胞因子在儿童支气管哮喘发病中发挥重要作用,并与哮喘的严重程度呈正相关。     

Th9和Th22细胞及其细胞因子在儿童支气管哮喘发病中的作用

目的检测支气管哮喘患儿外周血辅助性T细胞(Th)9和Th22细胞的比例以及血清中白细胞介素(IL)-9、IL-22和总IgE的表达,探讨其在儿童支气管哮喘发病中的作用及临床意义。方法选取56例哮喘患儿,其中哮喘发作组32例,哮喘缓解组24例。另选取20例健康儿童为对照组。采用流式细胞术(FCM)分别检测哮喘发作组、哮喘缓解组和健康对照组外周血中Th9细胞和Th22细胞的比例;应用酶联免疫吸附法(ELISA)检测各组儿童血清中IL-9和IL-22的含量;应用特定蛋白分析仪检测各组儿童血清中总IgE的水平。测得数据用SPSS 22.0统计软件进行分析,采用Pearson直线相关分析进行相关性检验。结果与哮喘缓解组和对照组相比,哮喘发作组外周血Th9和Th22细胞比例显著升高(P0.05);与对照组相比,哮喘缓解组外周血Th9和Th22细胞比例显著升高(P0.05)。与哮喘缓解组和对照组相比,哮喘发作组血清中IL-9、IL-22和总IgE水平显著升高(P0.05);与对照组相比,哮喘缓解组血清中IL-9、IL-22和总IgE水平显著升高(P0.05)。相关分析结果表明,支气管哮喘患儿血清中IL-9、IL-22与总IgE水平均呈正相关(P值均0.05)。结论支气管哮喘患儿外周血Th9和Th22细胞及其细胞因子在儿童支气管哮喘发病中发挥重要作用,并与哮喘的严重程度呈正相关。     

支气管哮喘患者外周血单个核细胞NLRP3mRNA和血清IL-18、IL-1β的表达和研究

目的检测支气管哮喘患者中外周单个核细胞Nod样受体热蛋白结构域相关蛋白3(NLRP3)mRNA和血清白细胞介素18(IL-18)、白细胞介素1β(IL-1β)的表达,探讨其在哮喘发病中的机制和临床意义。方法采用实时荧光定量聚合酶链反应(PCR)法检测100例支气管哮喘患者治疗前后和对照组外周血单个核细胞核内NLRP3mRNA的表达量,通过酶联免疫吸附测定(ELISA)法检测哮喘患者治疗前后和对照组血清IL-1β、IL-18水平,比较组间的差异,并进行相关性分析。结果急性发作期哮喘患者的NLRP3mRNA的表达及IL-18、IL-1β水平均明显高于慢性持续期哮喘患者(P0.05)。急性发作期和慢性持续期哮喘患者的NLRP3mRNA的表达及IL-18、IL-1β水平均明显高于健康对照组(P0.05)。急性发作期和慢性持续期哮喘患者经治疗后的NLRP3mRNA的表达量及IL-18、IL-1β水平较各自治疗前水平明显降低,且均高于健康对照组水平,差异具有统计学意义(P0.05)。结论 NLRP3、IL-18、IL-1β均参与了支气管哮喘的气道炎症反应,且与哮喘患者的病情严度程度密切相关。     

支气管哮喘患者IL-4、IL-5及血清IgE水平的变化

目的探讨支气管哮喘患者血清白细胞介素-4(IL-4)、白细胞介素-5(IL-5)、IgE水平及不同病程的差异.方法采用酶联免疫吸附试验(ELISA)检测了支气管哮喘患者经植物血凝素(PHA)刺激的外周血单介核细胞(PBMC)培养上清液中IL-4、IL-5水平,同时用散射比浊法进行IgE检测.结果支气管哮喘患者经PHA刺激的PBMC培养上清液中IL-4、IL-5及IgE较对照组明显增高,差异有显著性;哮喘发作期IL-4、IL-5、IgE水平较缓解期明显升高,差异有显著性.结论 IL-4、IL-5与支气管哮喘的发生密切相关.     

支气管哮喘患儿血清白细胞介素变化及其与 IgE、中性粒细胞百分比相关性分析

目的：通过观察支气管哮喘患儿血清中白细胞介素( IL-17)水平变化,分析IL-17在支气管哮喘发病过程中的作用。方法严格按照标准纳入80例急性发作期支气管哮喘患儿,以性别和年龄相匹配的40例门诊健康体检儿童作为健康对照组。采用酶联免疫吸附试验( ELISA)法测定支气管哮喘患儿及健康对照组血清中IL-17浓度;免疫比浊法测定免疫球蛋白E( IgE)。结果①支气管哮喘患儿血清IL-17中浓度高于健康对照组,差异具有显著性(P<0.05);②支气管哮喘患儿血清IL-17与IgE呈正相关(r=0.761,P<0.05);③支气管哮喘患儿血清IL-17与中性粒细胞百分比(NEU%)呈正相关(r=0.619,P<0.05)。结论IL-17与IgE有密切联系,共同参与了哮喘的发病。 IL-17通过促进中性粒细胞的增殖与募集,在支气管哮喘的发病过程中发挥着重要作用。     

C反应蛋白、白细胞介素8对哮喘病情评估价值研究

目的研究支气管哮喘患者急性发作期与非急性发作期血清c反应蛋白（cRP）、白细胞介素8（IL-8）水平变化,探讨血清CRP、IL-8水平对支气管哮喘急性发作期患者的诊断意义和临床价值,评价以上两种因子是否可成为支气管哮喘病情分期评估指标,以便更好地指导临床治疗。方法选择76例支气管哮喘急性发作期患者,经过规范的治疗,分别于治疗前、治疗后测定CRP、IL-8水平,同时选择68例健康成年人作为对照组;CRP、IL-8的检测分别应用免疫透射比浊法及酶联免疫吸附测定法。结果支气管哮喘急性发作期患者血清CRP含量明显高于健康对照组和非急性发作期,（52．8±6．2）mg／LVS（6．2±4．5）mg／L、（7．4±4．9）mg／L（均尸〈O．01）,而支气管哮喘非急性发作期患者与健康对照组比较差异无统计学意义;支气管哮喘急性发作期患者血清IL-8含量明显高于健康对照组和非急性发作期,（33．6±5．3）ng／LVS（19．6±5．1）ng／L、（24．9±4．9）ng／L（均P〈0．01）,支气管哮喘非急性发作期患者亦明显高于健康对照组（P〈0．01）。结论支气管哮喘患者急性发作期血清cRP、IL-8水平显著升高,提示血清CRP、IL-8水平检测可作为支气管哮喘急性发作期病情分期评估指标。     

支气管哮喘外周血单核细胞环氧化酶2的表达及临床意义

目的：检测支气管哮喘外周血单核细胞（PBMC）中环氧化酶-2（COX-2）mRNA的表达水平并探讨其意义。方法：应用逆转录-聚合酶链反应（RT-PCR）检测了38例支气管哮喘患者和20例正常人PBMC中COX-2的表达水平。结果：支气管哮喘发作期患者PBMC中COX-2的平均表达水平（0.82±0.13）明显高于缓解期（0.20±0.31）和正常对照组（0.17±0.06）,差异非常显著（P〈0.01）;哮喘缓解期患者PBMC中COX-2的平均表达水平与正常对照组间差异无显著性（P〉0.05）。结论：COX-2表达水平可用以评估支气管哮喘的炎症状况。     

哮喘患儿外周血RORC mRNA、IL-17水平及其与气道重塑的关系研究

目的：研究Th17细胞上游调控基因维甲酸相关孤儿核受体C （RORC）mRNA及主要效应分子IL-17在支气管哮喘患儿外周血单个核细胞中的表达变化,并探讨其与哮喘气道重塑的关系。方法收集2010年9月至2012年8月中山大学孙逸仙纪念医院儿科就诊的中重度持续哮喘儿童45例,其中哮喘急性发作期25例,临床缓解期20例,以20名健康儿童为对照组。采用荧光实时定量PCR技术和双抗体夹心ELISA法分别检测PBMC中 RORC mRNA和IL-17的表达;选取其中14例病程较长的临床缓解期患儿,采用高分辨螺旋CT进行胸部检测及支气管壁测量,并与同期IL-17水平进行相关分析。结果哮喘儿童急性发作期外周血PBMC 表达RORC mRNA、IL-17水平均显著高于缓解组和正常对照组;缓解期患儿IL-17水平比对照组明显增高（P＜0．05）,而RORC mRNA表达水平与对照组比较差异无统计学意义（P＞0．05）。中重度哮喘患儿气道壁厚度、气道壁面积均明显增加,且与IL-17水平呈正相关（r 分别为0．78、0．77,P＜0．05）。结论转录因子RORC及IL-17参与儿童哮喘发病,其中IL-17水平升高与儿童哮喘气道重塑密切相关。     

Expression of mRNA for interleukin-5 in mucosal bronchial biopsies from asthma.

We have attempted to identify mRNA for IL-5 in endobronchial mucosal biopsies from asthmatics and controls, using the technique of in situ hybridization. Bronchial biopsies were obtained from 10 asthmatics and 9 nonatopic normal controls. A radio-labeled cRNA probe was prepared from an IL-5 cDNA and hybridized to permeabilized sections. These were washed extensively before processing for autoradiography. An IL-5-producing T cell clone derived from a patient with the hyper-IgE syndrome was used as a positive control. As a negative control, sections were also treated with a "sense" IL-5 probe. Specific hybridization signals for IL-5 mRNA were demonstrated within the bronchial mucosa in 6 out of the 10 asthmatic subjects. Cells exhibiting hybridization signals were located beneath the epithelial basement membrane. In contrast, there was no hybridization in the control group. No hybridization was observed with the sense probe. The six IL-5 mRNA-positive asthmatics tended to have more severe disease than the negative asthmatics, as assessed by symptoms and lung function, and showed a significant increase in the degree of infiltration of the bronchial mucosa by secreting (EG2+) eosinophils and activated (CD25+) T lymphocytes. Within the subjects who showed positive IL-5 mRNA, there was a correlation between IL-5 mRNA expression and the number of CD25+ and EG2+ cells and total eosinophil count. This study provides evidence for the cellular localization of IL-5 mRNA in the bronchial mucosa of asthmatics and supports the concept that this cytokine regulates eosinophil function in bronchial asthma.     

The role of interleukin-4 and interleukin-13 in the non-immunologic aspects of asthma pathogenesis.

Bronchial asthma is a complex disease characterized by airway inflammation involving a Th2-cytokine, interleukin (IL)-13. A substantial body of evidence has accumulated pointing to the pivotal role of IL-13 in the pathogenesis of bronchial asthma. The evidence is categorized as (i) analyses of mouse models, (ii) expression of these cytokines in the bronchial lesions, and (iii) genetic association of the signaling molecules of these cytokines. In addition, the molecular mechanism of the signal transduction of IL-13 has also been well characterized. We have applied microarray analyses to human bronchial epithelial cultures to search for genes regulated by IL-13 and have identified a subset of disease-relevant genes by comparison with cDNA libraries derived from normal and asthmatic bronchial biopsies. Expression of squamous cell carcinoma antigen-1 (SCCA1) and SCCA2, the cysteine and serine protease inhibitors, respectively, was the highest in the bronchial epithelial cells stimulated by IL-4 and IL-13 and was augmented in the asthmatic cDNA library. Furthermore, serum levels of SCCA were also elevated in asthmatic patients. Taken together, it was supposed that SCCA may play some role in the pathogenesis of bronchial asthma, and measuring its serum level may be relevant for diagnosing or monitoring the status of bronchial asthma.     

双歧杆菌对过敏性哮喘儿童DC成熟及其分泌细胞因子的影响

目的研究青春型双歧杆菌对过敏性哮喘儿童外周血单个核细胞（PBMC）来源的树突状细胞（DC）表达CD86和HLA-DR及其分泌IL-1βI、L-6I、L-10、IL-12、IL-23和IFN-γ的影响。方法从15名过敏性哮喘儿童和15名非哮喘儿童的外周血单个核细胞诱导生成未成熟DC,分别加入青春型双歧杆菌或脂多糖（LPS）后继续培养DC 2天,①每天于倒置显微镜下观察DC形态;②用流式细胞仪检测各组DC表面CD86和HLA-DR的分子表达;③用ELISA方法检测培养上清中IL-1βI、L-6I、L-10、IL-12I、L-23和IFN-γ的水平。结果①DC经双歧杆菌和LPS分组处理后第2天,于倒置显微镜下观察到双歧杆菌组和LPS组的DC突起均较空白组明显,具有突起的细胞数量较空白组增多,而双歧杆菌组与LPS组比较,LPS组有突起的细胞数量明显增多,提示DC过度生长。②双歧杆菌刺激后,哮喘儿童DC表面CD86表达明显增高,HLA-DR表达无明显变化,非哮喘儿童CD86和HLA-DR表达均无明显变化;LPS刺激可明显增加哮喘儿童和非哮喘儿童CD86和HLA-DR的表达。③双歧杆菌刺激哮喘儿童DC分泌IL-12、IFN-γI、L-1β及IL-6水平增高,刺激非哮喘儿童DC分泌IL-12I、L-10I、L-1β及IL-23水平增高。结论①过敏性哮喘儿童DC表面CD86的表达可能存在缺陷,双歧杆菌能适度上调其表达,在DC的成熟过程中可能起调节作用。②双歧杆菌能刺激过敏性哮喘儿童DC分泌IL-12和IFN-γ,从而改变Th2优势分化,纠正Th1/Th2失衡。③过敏性哮喘儿童DC分泌IL-10可能存在缺陷,从而影响免疫耐受的形成。④双歧杆菌可能通过刺激哮喘儿童DC分泌IL-1β及IL-6增高,达到促进Th17细胞分化的作用。     

Comparative assessment of various methods of determination of interleukin-5 production and their clinical significance in bronchial asthma

Interleukin-5 (IL-5) is an important marker of inflammation in bronchial asthma (BA). The level of IL-5 was investigated by immune-enzyme assay (IEA); the expression degree of IL-5 mRNA was studied, before and after the conducted therapy, by the inhibition reaction-IEA (IR-IEA) in sputum and blood serum of patients. No differences between contents of IL-5 were found in blood plasma of patients with various disease degrees or of patients with different BA etiologies. The IL-5 contents in sputum were reliably different in different groups and depended on a disease severity, exacerbation and remission. An evaluation of an expression degree of the IL-5 RNAm in eosinophiles, derived from patients' blood, provided for clarifying the differences between acute asthma and other disease forms and for defining the therapy influence on the parameter in question. The IL-5 RNAm expression in sputum was reliably different in patients with moderate forms and with acute forms of the disease; it was decreasing due to treatment. Finally, the results of the evaluation of the IL-5 level and the study of mRNA expression of the cytokine mutually supplement one another and make it possible to evaluate the disease degree and therapy efficiency.     

Reaction of mononuclear cells from patients with bronchial asthma to Candida albicans

This study was undertaken to investigate cytokine production of mononuclear cells (MNCs) from patients with bronchial asthma stimulated by antigens of Candida albicans in vitro. The reaction of MNCs from corticosteroid-dependent patients was restricted to a low level. The level of tumor necrosis factor (TNF)-alpha released into the culture fluid was significantly higher in the high responders (HR) of the atopic corticosteroid-independent (A-CSID) group than those of other asthmatic and control groups (p less than 0.01). The level of IL-1 beta of the A-CSID group was significantly higher than that of the non-atopic corticosteroid-independent group (p less than 0.05), but not significantly different from the controls. In HR of the A-CSID group, the production of TNF-alpha and IL-1 beta was augmented and interferon-gamma production was also increased in these patients. These results suggest that Candida albicans can contribute to the augmentation of cytokine production in bronchial asthma, especially in some of atopic type.     

白细胞介素类mRNA在儿童哮喘中异常表达的检测及意义

目的：研究白细胞介素－12（IL－12），IL－13在哮喘发病中的作用。方法：用逆转录聚合酶链反应（RT－PCR）法半定量分析了哮喘急性发作期及正常对照组儿童外周血单个核细胞（PBMC）中IL－12，IL－13mRNA表达水平的变化，同时对IgE水平进行了检测，结果：哮喘组IL－12 mRNA表达减少，IL－13 mRNA表达增多，病情越重，IL－12mRNA表达越少，IL－13 mRNA表达越多，无论IgE升高与否。与对照组比较，IL－12，IL－13mRNA表达，差异均有显著性，结论：IL－12，IL－13可能是构成气道慢性炎症的各类因素之一。     

血液透析对终末期肾病患者体内IL-13水平的影响

目的探讨终末期肾病（End stage renal disease ESRD）患者血浆白细胞介素13（Interleukin 13 IL-13）水平以及血液透析（Hemodialysis HD）对其的影响，并判断其能否作为评价透析膜生物相容性的指标。方法选择同济大学附属东方医院肾内科30例ESRD患者分为未透析组和维持性血液透析、（Maintenance hemodialysiS MHD）组，MHD组随机分为纤维素生物膜组（650）和聚砜膜组（F6）各10例，同时设立健康正常者10例为对照组。应用酶联免疫吸附（ELISA）法测定MHD患者透析前、后血浆中IL-13水平，以及外周血单个核细胞（Peripherial blood mononuclear cell SPBMC）经及未经植物凝集素（Phytohemagglutinin PHA）干预的培养上清中IL-13的水平，同时以半定量逆转录多聚酶链反应（RT-PCR）法检测PBMC中IL-13mRNA的表达。结果ESRD未透析组及MHD组透析前血浆IL-13水平较正常对照明显升高，MHD组经透析后血浆IL-13水平下降。ESRD未透析患者的PBMc经PHA刺激后IL-13培养上清水平及mRNA表达量均明显降低，650组PBMC经PHA刺激后IL-13培养上清水平及mRNA表达量无明显变化，F6组PBMC经PHA刺激后IL-13培养上清水平及mRNA表达量明显升高。结论①ESRD患者体内处于微炎症状态；②血液透析可改善其炎症状态，从而使IL-13水平下调；③聚砜膜较纤维素生物膜能更好的改善患者的T细胞对抗原的反应能力；④IL-13可作为评价透析膜生物相容性的指标之一。