咳嗽变异性哮喘患者诱导痰中神经生长因子和白细胞介素-4水平及气道炎症特征初探

目的 通过观察咳嗽变异性哮喘(CVA)患者诱导中神经生长因子(NGF)和IL-4水平,初步探讨咳嗽变异性哮喘的气道炎症特征.方法 选咳嗽变异性哮喘患者36例及健康体检者23例,对受试者进行痰诱导,查诱导痰中细胞分类计数,酶联免疫吸附法检测诱导痰中NGF和IL-4水平.结果 (1)咳嗽变异性哮喘患者诱导痰中嗜酸性粒细胞百分数为8%,显著高于健康体检者(1%),差异有统计学意义(P＜0.001).其中13例患者应用糖皮质激素联合长效β2受体激动剂(布地奈德/福莫特罗,每吸160μg/4.5μg,2吸/d)治疗1个月后,咳嗽症状明显好转,诱导痰中嗜酸性粒细胞百分数为2%,显著低于治疗前(5%),差异有统计学意义(P＜0.05);但仍高于健康体检者.(2)咳嗽变异性哮喘患者诱导痰中NGF和IL-4浓度高于健康体检者,差异有统计学意义(P＜0.05).其中13例患者经上述治疗后诱导痰中NGF和IL-4浓度下降,差异有统计学意义(P＜0.05);但仍高于健康体检者.(3)相关性分析:诱导痰中嗜酸性粒细胞计数与诱导痰上清中NGF、IL-4浓度呈正相关(r分别为0.397、0.332,P＜0.01).诱导痰上清中NGF与IL-4浓度呈正相关(r=0.728,P＜0.01).结论 神经-免疫机制与咳嗽变异性哮喘嗜酸性粒细胞性炎症密切相关,NGF和IL-4参与并介导了这一炎症.糖皮质激素联合长效β2受体激动剂吸入治疗,能显著降低诱导痰中NGF、IL-4和嗜酸性粒细胞水平,减轻嗜酸性粒细胞性炎症.

Abstract：

Objective To observe sputum cytology counts, the levels of nerve growth factor (NGF) and IL-4 in cough variant asthma (CVA) patients and the change of their levels after using glucocorticoids combined with β2-adrenergic agonists one month, and to investigate CVA's characteristics of airway inflammation. Methods Totally 36 patients with untreated CVA were selected, as well as 23 healthy controls. Coughed up sputum cells were obtained and HE strained for differential cell counting in each enrolled patient. In induced sputum's supernatant, the levels of NGF and IL-4 were determined by ELISA.Results Before treatment, CVA patients had a median eosinophils (EOS) percentage of 8%, which was significantly higher than that after treatment (2%, P＜0.05) and in healthy control group (1%, P＜0. 001). The levels of NGF and IL-4 in induced sputum of CVA group were (9. 50 ± 1.69) ng/L and (257.37 ± 53.57) ng/L. After treatment, they were (8.78±1.02) ng/L and (228.60 ±52.93)ng/L in CVA group, (6.98±0.69) ng/L and (166.44±24.75) ng/L in healthy control group. The levels of NGF and IL-4 before and after treatment in the CVA group , as compared with the healthy control group, had statistically significant differences (all P＜0.001). In CVA group before and after treatment, the level of NGF and IL-4 paired difference was significant (P＜0.001). The percentage of induced sputum EOS correlated with sputum supernatant concentrations of NGF and IL-4 (P ＜ 0.01). In induced sputum supernatant, the concentrations of NGF and IL-4 were significant correlated (P＜0.01). Conclusions Glucocorticoid joint long-term β2 agonist inhaled treatment significantly reduced NGF, IL-4 and EOS levels and reduced eosinophilic inflammation, which are closely related with the nerve-immune mechanism, NGF as well as IL-4 participated the inflammation. Induced sputum examination is non-invasive, economical,simple, easily accepted by patients, and repeatable, widely used in clinical.     

支气管哮喘患者诱导痰中基质细胞衍生因子的表达及其作用

目的 测定支气管哮喘(简称哮喘)患者在糖皮质激素(简称激素)治疗前后诱导痰中基质细胞衍生因子-1(SDF-1)和白细胞介素(IL)-17的水平,探讨SDF-1在哮喘发病机制中的作用.方法 收集2009年6月至2010年9月郑州大学第一附属医院门诊及住院的慢性持续期的哮喘患者99例(按病情严重程度分为轻、中、重度组)及健康体检者30名,哮喘患者在回答哮喘控制问卷(ACQ)后,两组研究对象分别进行肺功能检测和诱导痰检查,记录FEV1占预计值%,行诱导痰炎症细胞分类计数,酶联免疫吸附试验(ELISA)法检测诱导痰中SDF-1和IL-17水平;所有哮喘患者均参照支气管哮喘指南给予规范的吸入激素为主的治疗,4周后测定诱导痰中SDF-1、IL-17的水平及炎症细胞比率.结果 轻、中、重度持续组的ACQ评分及FEV1占预计值%差异均有统计学意义(F值分别为79.271和457.448,均P<0.01).治疗前哮喘组诱导痰嗜酸粒细胞比率、IL-17水平及SDF-1水平均高于健康对照组(均P<0.01);重度哮喘患者诱导痰中两种炎症细胞比率及SDF-1和IL-17水平均高于轻度哮喘患者(均P<0.05).哮喘患者FEV1占预计值%与诱导痰中嗜酸粒细胞、中性粒细胞比率均呈负相关(r值分别为-0.316和-0.409,均P<0.05);诱导痰中SDF-1与嗜酸粒细胞比率及中性粒细胞比率呈正相关(r值分别为0.875和0.716,均P<0.01);诱导痰中IL-17与嗜酸粒细胞及中性粒细胞比率呈正相关(r值分别为0.878和0.846,均P<0.01);诱导痰中SDF-1与IL-17水平呈正相关(r=0.872,P<0.01).治疗后哮喘患者诱导痰中两种炎症细胞比率及SDF-1和IL-17水平均低于治疗前患者(均P<0.01);治疗后未控制组哮喘患者诱导痰中性粒细胞比率、SDF-1水平和IL-17水平明显高于完全控制组(均P<0.05).结论 SDF-1和IL-17通过募集炎症细胞,特别是中性粒细胞参与了哮喘气道炎症的发生,SDF-1可作为哮喘患者临床病情判定和疗效观察的参考指标.

Abstract：

Objective To evaluate concentrations of stromal cell-derived factor 1 (SDF-1) and IL-17 in induced sputum supernatants from asthmatic patients before and after treatment with glucocorticosteroids. Methods Induced sputum was collected from 30 healthy controls and 99 patients with chronic persistent asthma from 2009-2010. Sputum samples were obtained before and after 4 week treatment with inhaled glucocorticosteroids. The sputum concentrations of SDF-1 and IL-17 were measured by ELISA. Results The FEV1% and the asthma control score of patients with severe asthma were decreased as compared with patients with moderate persistent and mild persistent asthma (F=457.448 and 79.271, all P<0.01). The concentrations of SDF-1 ,IL-17 and the percentage of eosinophils were increased in asthma group compared with control subjects (all P<0.01),but the percentage of sputum neutrophils was lower than that in the healthy controls(P<0.01). The percentage of sputum neutrophils and eosinophils and the level of SDF-1 and IL-17 in patients with severe persistent asthma were significantly higher than those in patients with mild persistent asthma (all P<0.05). The percentage of sputum neutrophils and eosinophils were negatively correlated with FEV1%(r=-0.409 and -0.316,all P<0.05). The levels of IL-17 and SDF-1 were positively correlated with the percentage of sputum neutrophils and eosinophils (all P<0.01). The levels of IL-17 were positively correlated with the levels of SDF-1(r=0.872, P<0.01).After glucocorticosteroid therapy, the percentage of eosinophils and neutrophils, the levels of IL-17 and SDF-1 decreased significantly in all patients(all P<0.01), while the percentage of sputum neutrophils and the levels of IL-17and SDF-1 in uncontrolled patients increased significantly compared with the controlled and partly controlled groups(all P<0.05). Conclusions SDF-1 and IL-17 may contribute to airway inflammation in asthma by chemotactic activity towards neutrophils. The concentration of SDF-1 may be used to evaluate the inflammation and the therapeutic effects.     

甲泼尼龙对慢性咳嗽患者诱导痰巨噬细胞中Toll样受体2表达的影响

目的 探讨Toll样受体2(TLR2)在慢性咳嗽患者诱导痰巨噬细胞中的表达与气道炎症的关系以及甲泼尼龙对TLR2表达的影响.方法 收集2008年9月至2010年3月泸州医学院附属医院呼吸科门诊的慢性咳嗽患者86例,其中咳嗽变异性哮喘(CVA)组31例,上气道咳嗽综合征(UACS)组14例,嗜酸细胞性支气管炎(EB)组16例,胃食管反流性咳嗽(GERC)组25例.对照组为20名健康志愿者.各组均行痰液诱导检查,HE染色,免疫荧光染色测定TLR2表达,酶联免疫吸附法(ELISA)测定诱导痰上清液中细胞因子的浓度.结果 (1)CVA、UACS、EB和GERC各组患者诱导痰中细胞总数[分别为(3.4±1.1)×106/L、(2.6±0.6)×106/L、(2.6±0.6)×106/L、(2.7±0.5)×106/L]均明显高于对照组[(1.4±0.6)×106/L],差异均有统计学意义(均P＜0.01);分类计数显示以上4组痰中巨噬细胞比例(分别为0.35±0.04、0.40±0.06、0.38±0.04、0.43±0.05)均明显低于对照组(0.63±0.08);中性粒细胞比例(分别为0.52±0.06、0.58±0.06、0.52±0.04、0.54±0.06)明显高于对照组,差异均有统计学意义(均P＜0.01).CVA和EB组患者嗜酸细胞比例(分别为0.115±0.054和0.099±0.034)明显高于其他各组,差异有统计学意义(均P＜0.01),但两组间差异无统计学意义(t=1.18,P＞0.05).(2)CVA、UACS、EB和GERC各组患者诱导痰上清液白细胞介素-8浓度[分别为(7.0±1.2)μg/L、(10.2±3.1)μg/L、(6.3±1.9)μg/L、(7.7±2.5)μg/L]和肿瘤坏死因子-α浓度[分别为(30±11)ng/L、(46±16)ng/L、(2l±5)ng/L、(33±15)ng/L]明显高于对照组[分别为(1.6±0.8)μg/L和(13±6)ng/L],差异均有统计学意义(均P＜0.01).(3)CVA、UACS、EB和GERC各组患者诱导痰上清液中白细胞介素-8与痰中中性粒细胞数呈正相关(r值分别为0.628、0.816、0.769、0.733,均P＜0.05);CVA、UACS、EB和GERC各组患者诱导痰上清液中肿瘤坏死因子-α与痰中中性粒细胞数呈正相关(r值分别为0.579、0.865、0.730、0.755,均P＜0.05).(4)CVA、UACS、EB和GERC各组患者诱导痰巨噬细胞TLR2表达量(56±15、38±7、65±17、27±4)明显低于对照组(135±14),甲泼尼龙可上调以上各组患者诱导痰巨噬细胞中TLR2(75±17、53.29±16、94±30、41±5)的表达.结论 CVA、UACS、EB和GERC等慢性咳嗽患者诱导痰巨噬细胞中TLR2的表达下降,甲泼尼龙可上调巨噬细胞TLR2的表达.

Abstract：

Objective To explore the relationship between airway inflammation and Toll-like receptor 2 ( TLR2 ) expression on macrophages in induced sputum from chronic cough patients, and to observe the effect of methylprednisolone on this receptor. Methods Eighty-six outpatients with chronic cough were enrolled in this study from the respiratory department in the Affiliated Hospital of Luzhou Medical College from September 2008 to March 2010. These diseases were diagnosed according to the guideline of Chinese Society of Respiratory Diseases in 2005, excluding severe liver, heart and kidney pathology and pregnant women. Among 86 cases, 31 were cough variant asthma ( CVA), 14 upper airway cough syndrome (UACS), 16 eosinophilic bronchitis (EB) and 25 gastroesophageal reflux (GERC). Twenty healthy volunteers were included as controls. Induced sputum was collected and cell counts were analyzed. TLR2 expression on macrophages was determined by immunofluorescence staining. Cytokine levels were measured with enzyme linked immunosorbent assay (ELISA). Results (1) The total inflammatory cells [(3.4±1.1) ×106/L, (2.6 ±0.6)×106/L, (2.6±0.6)×106/L, (2.7±0.5) × 106/L] and neutrophils (0.52±0.06, 0.58 ±0.06, 0.52±0.04, 0.54 ±0.06) in induced sputum from CVA, UACS, EB and GERC were significantly increased(t value = 5.27, 4.09, 3. 16, 3.92 and 3.62, 3.49, 4.82, 6.17respectively, all P＜0.01 ), with decreased macrophages (0.35±0.04, 0.40±0.06,0.38±0.04,0.43 ±0. 05) in comparison with the control group (t value =4. 76,5.24,4. 57,3. 18, all P ＜0. 01 ). There was a significant increase in eosinophils(0.118±0.054,0.099±0.034) in CVA and EB group (t value =4.46,3.87,5. 19 and 3.51,4. 06,4. 38 respectively, all P＜0.01). (2) The concentration of IL-8[(7.0±1.2)μg/L,(10.2±3.1)μg/L,(6.3±1.9)μg/L,(7.7±2.5)μg/L] and TNF-α[(30 ± 11)ng/L, (46±16)rng/L, (21 ±5)ng/L, (33 ± 15)ng/L] from induced sputum in CVA, UACS, EB and GERC groups were significantly increased, compared with the control group( t value =4. 58,3.67,4. 26,5. 39 and 5.16,3.97,4. 59,3.34 respectively, all P ＜ 0.01 ). (3) The concentration of IL-8 was positively correlated with neutrophils in each group( R value = 0.628,0. 816,0. 769 and 0. 733 respectively, all P＜0. 05 ). The concentration of TNF-o was also positively correlated with neutrophils in each group ( R value = 0. 579,0. 865,0. 730 and 0. 755 respectively,all P ＜ 0. 05 ). (4) The expression of TLR2 ( 56 ± 15,38 ± 7,65 ±17,27 ±4) on macrophages in CVA, UACS,EB and GERC was significantly decreased. The expression of TLR2 (75±17, 53±16,94±30,41±5 ) on macrophages was enhanced by methylprednisolone.Conclusions The expression of TLR2 on macrophages from induced sputum in CVA, UACS, EB and GERC was decreased, but could be enhanced by methylprednisolone, suggesting that TLR2 might play a regulatory role in airway inflammatory response.     

呼出气一氧化氮测定在慢性咳嗽诊治中的应用价值初探

目的 评估呼出气一氧化氮(FENO)在慢性咳嗽诊断和治疗中的临床应用价值.方法 根据中华医学会呼吸病学分会哮喘学组制定的咳嗽诊断与治疗指南(2009版),纳入卫生部中日友好医院呼吸内科门诊2010年1-6月明确诊断为慢性咳嗽的连续病例75例,其中咳嗽变异性哮喘(CVA组)29例,嗜酸粒细胞性支气管炎(EB组)19例,其他病因慢性咳嗽(胃食管反流性咳嗽、上气道咳嗽综合征等)27例,对所有患者分别进行FENO检测、诱导痰细胞分类计数、肺功能及激发试验、血清总IgE检测等,并评价咳嗽症状积分、生活质量问卷.按指南推荐的方案治疗4周后随访,再次进行上述评价.结果 CVA组的FENO值为(58±26)ppb,高于EB组的(36±18)ppb及其他慢性咳嗽组的(20±7)ppb,EB组高于其他慢性咳嗽组(F=28.2,P<0.01).相关性分析结果显示,治疗前慢性咳嗽患者的FENO与诱导痰嗜酸粒细胞比例(r=0.56)、与激发试验传导率下降的斜度(r=0.57)、与咳嗽症状总积分(r=0.61)均显著相关(均P<0.01).与血清总IgE水平、咳嗽生活质量问卷总分也具有相关性.CVA、EB组患者抗炎治疗后咳嗽缓解患者的基线FENO值为(63±42)ppb,高于未缓解患者的(28±13)ppb(t=3.91,P<0.01).抗炎治疗后FENO下降的比例与痰嗜酸粒细胞下降比例显著相关(r=0.53,P<0.01),与咳嗽症状积分下降比例显著相关(r=0.48,P<0.01).结论 呼出气一氧化氮与慢性咳嗽患者的嗜酸性气道炎症、气道反应性、特应质及咳嗽症状均存在相关性,在慢性咳嗽的诊治中具有一定的辅助价值.

Abstract：

Objective To evaluate the values of fractional exhaled nitric oxide (FENO) in the diagnosis and treatment of chronic cough. Methods Based on the protocol from The Chinese Respiratory Society guidelines for management of cough, 75 consecutive subjects with chronic cough and normal chest radiographs were recruited from the outpatient clinic of the Department of Respiratory Diseases in China-Japan Friendship Hospital from January to June 2010. All the patients accepted FENO tests, sputum cell counts, pulmonary function tests, bronchial hyperresponsiveness(BHR), serum IgE, cough symptom scores and Licester cough Questionnaire (LCQ) before and after treatment of 4 weeks. Results The final diagnosis of the subjects included 29 with cough variant asthma (CVA), 19 with eosinophilic bronchitis (EB) and 27 with other causes(Others). FENO levels in CVA(58±26)ppb were significantly higher than those in EB(36±18)ppb and Others(20±7)ppb, and the FENO levels in EB were higher than Others(F=28.2,P<0.01). FENO levels showed significant correlations with sputum eosinophils, BHR, cough symptom scores, non-specific IgE, and LCQ scores. The mean baseline FENO level of the subjects whose coughs were relieved after inhaled corticosteroid therapy was(63±42)ppb, higher than those of the non-responders (28±13)ppb(t=3.91,P<0.01). There were significant correlations between the percentage of FENO decrease and the percentage of sputum eosinophil decrease or the cough symptom score decrease. Conclusion FENO could be used as a inflammation marker of chronic cough because of its good correlation with sputum eosinophils, AHR, atopy, and cough symptoms. FENO also has a potential to predict the response of anti-inflammatory therapy because FENO decrease is correlated with the decrease of eosinophilic inflammation and improvement of cough symptoms.     

Airway remodeling assessed by high-resolution computed tomography in patients with asthma:relationship to biological markers in induced sputum

Objective To explore the significance of assessing asthma control by high-resolution computed tomography(HRCT) and biological markers in induced sputum.Methods Forty-eight patients with asthma(asthma group) and 10 healthy subjects(control group) were retrospectively analyzed.     

Effect of electro-acupuncture at Foot-Yangming Meridian on somatostatin and expression of somatostatin receptor genes in rabbits with gastric ulcer

AIM: To discuss the protective effect of electroacupunc-ture at the Foot-Yangming Meridian on gastric muco-sal lesion, somatostatin (SS) and the expression of SS receptor genes (SSRimRNA ) in rabbits with gastric ulcer and to further explore the relative specificity of meridians and viscera at gene expression level. METHODS: Forty rabbits were randomly divided into control group (A), gastric ulcer model group (B), Foot-Yangming Meridian group (C), Foot-Shaoyang Meridian group (D) and Foot-Taiyang Meridian group (E). The gastric ulcer model was prepared by infusing alcohol into stomach. Groups C-E were treated with electro-acupuncture at points along the above meridians using meridian stimulating instruments for 7 days respectively. By the end of treatment, the index of gastric ulcer was determined, the amount of epidermal growth factor(EGF) and somatostatin was measured by radioimmunoassay (RIA). SS-RimRNA expression in gastric mucosa was determined by RT-PCR. RESULTS: The value of EGF in model group was obviously lower(73.6±14.8 vs 91.3±14.9 pg/mL, P<0.01) than that in control group. The index of gastric ulcer, content of SS and expression of SSRimRNA in gastric mucosa were significantly higher than those in control group (24.88±6.29 vs 8.50±2.98 scores, P<0.01; 2978.6±587.6 vs 1852.4±361.7 mIU/mL, P<0.01; 2.56±0.25 vs 1.04±0.36, P<0.01) . The value of EGF in Foot-Yangming Meridian group was higher than that in model group(92.2±6.7 vs 73.6±14.8 pg/mL, P<0.01). The index of gastric ulcer, content of SS and expression of SS-R1mRNA in gastric mucosa were significantly lower than those in control group(10.88±3.23 vs 24.88±6.29 scores, P< 0.01; 1800.2±488 vs 2978.6±587.6 mIU/mL, P<0.01; 1.07±0.08 vs 2.56±0.25mIU/mL, P<0.01). Compared to the model group, the content of SS and expression of SSRimRNA in gastric mucosa in Foot-Shaoyang Meridian group decreased (2441.0±488. vs 2978.6±587.6 mIU/mL, P<0.05;1.73±0.16 vs 2.56±0.25 mIU/mL, P<0.01). But the above parameters in Foot-Taiyang Meridian group did not improve and were significantly different from those in Foot-Yangming Meridian group (P<0.05) CONCLUSION: Electro-acupuncture at Foot-Yangming Meridian can protect gastric mocusa against injury. The mechanism may be releted to the regulation of brain-gut peptides and the expression of SSRimRNA.     

Serum sIL-2R,TNF-α and IFN-γ in alveolar echinococcosis

AIM:To approach the relationship between alveolarechinococcosis (AE) pathology and level of sIL-2R,TNF-αand IFN-γ in sera and the significance of cytokines indevelopment of AE.METHODS:After 23 patients with AE were confirmed byELISA and ultrasound,their sera were collected and theconcentrations of sIL-2R,TNF-α and IFN-γ were detectedby double antibody sandwich.Twelve healthy adults servedas controls.According to the status of livers of AE patients byultrasound scanning,they were divided into 4 groups:P_2,P_3,P_4 groups and C group (control).Average of concentrationsof sIL-2R,TNF-α and IFN-γin homologous group was statisticallyanalyzed by both ANOV and Newman-Keuls,respectively.RESULTS:The mean of sIL-2R in P_2 group was 97±29,P_3:226±80,P_4:194±23 and control group (111±30)×10~3 u/L(P<0.01).The mean of TNF-α in P_2 group was 1.12±0.20,P_3:3.67±1.96,P_4:1.30±0.25 and control group 0.40±0.19 μg/L(P<0.01).The mean of IFN-γ in P_2 group was 360±20,P_3:486±15,P_4:259±19 and control group:16±2 ng/L (P<0.01).Judged by ANOV and Newman-Keuls,the mean concentrationsof sIL-2R,TNF-α and IFN-γ had a significant differenceamong groups.Except for P_2 group,the mean sIL-2R betweenother groups of AE patients had a significant difference(P<0.05).The mean of TNF-α concentration in P_3 groupwas the highest (P<0.01).The mean of IFN-γ concentrationin all patients was higher than that in control group (P<0.01),but there was no difference between AE groups (P>0.05).CONCLUSION:Low sIL-2R level indicates an early stage ofAE or stable status,per contra,a progression stage.Higherlevel of TNF-α might be related to the lesion of liver.Therote of single IFN-γ is limited in immunological defenseagainst AE and it can not fully block pathological progression.     

嗜酸粒细胞性支气管炎患者气道炎症细胞及介质特征的探讨

目的观察嗜酸粒细胞性支气管炎(EB)诱导痰和支气管肺泡灌洗液(BALF)中细胞分类和炎症介质浓度,探讨EB的气道炎症特征。方法对43例EB(EB组)患者行诱导痰检查,将20例咳嗽变异型哮喘(CVA)患者(CVA组)、16例典型支气管哮喘(哮喘组)患者和21名健康人(健康对照组)行对照诱导痰检查,并对部分EB(11例)和CVA患者(10例)行支气管肺泡灌洗(BAL)。观察检测诱导痰、BALF中的细胞分类、嗜酸粒细胞阳离子蛋白(ECP)、白三烯C4(LTC4)和组胺的浓度。结果EB组患者诱导痰中嗜酸粒细胞(EOS)百分比为0.1130±0.1470,CVA组为0.1900±0.1800,哮喘组为0.3860±0.2670,与健康对照组(0.0020±0.0050)比较差异有统计学意义(P均<0.01);哮喘组与CVA组、CVA组与EB组比较差异均有统计学意义(P均<0.05);EB组BALF中EOS为0.011±0.016,CVA组为0.053±0.040,两组比较差异有统计学意义(P<0.05);EB组诱导痰中的ECP浓度为(0.62±0.66)mg/L、CVA组为(1.27±1.74)mg/L,对照组为(0.07±0.10)mg/L,3组间比较差异有统计学意义(P<0.01);CVA组诱导痰中的LTC4浓度为(0.65±0.62)μg/L,EB组为(0.39±0.61)μg/L,对照组为(0.15±0.11)μg/L,3组间比较差异有统计学意义(P分别<0.05、0.01);CVA组BALF中组胺浓度为(3.4±1.4)μg/L,EB组为(1.6±1.5)μg/L,两组比较差异有统计学意义(P<0.05)。结论EB组EOS炎症主要局限于中心气道,部分气道炎性介质水平低于CVA组。上述气道炎性特征可能是EB患者无非特异性气道高反应性的重要机制。     

支气管刷检嗜酸粒细胞计数在嗜酸粒细胞性支气管炎诊断中的价值

目的探讨支气管刷检嗜酸粒细胞（EOS）计数在嗜酸粒细胞性支气管炎（EB）诊断中的价值。方法选择32例EB患者（EB组）,18例咳嗽变异性哮喘患者（CVA组）,26例其它病因咳嗽患者（其它病因组）和13名健康人（健康对照组）,分别进行诱导痰、支气管刷检洗涤液和支气管肺泡灌洗液（BALF）中EOS的检测。结果EB及CVA组各标本中EOS的比例较其它病因及健康对照组明显增高（P〈0．001）;在EB及CVA组,支气管刷检洗涤液中EOS的比例显著高于诱导痰和BALF（P均〈0．叭）;支气管刷检EOS计数诊断EB的敏感性、特异性、阳性预测值和阴性预测值分别为100％、71．9％、66．7％和100％。结论经纤支镜支气管刷检EOS计数在EB的诊断中具有较重要的价值。     

吸入糖皮质激素(布地奈德)/长效β2受体激动剂(福莫特罗)复合制剂治疗中、重度持续支气管哮喘的疗效研究

目的 观测联合吸入糖皮质激素(布地奈德)/长效β2受体激动剂(福莫特罗)干粉吸入剂治疗期间支气管哮喘(简称哮喘)患者诱导痰嗜酸粒细胞(EOS)计数、肺功能与哮喘临床症状的关系.方法 本院健康呼吸中心就诊的中、重度持续哮喘患者33例(哮喘组)联合吸入糖皮质激素/长效β2受体激动剂复合制剂治疗6个月,测定肺功能(FEV1、FEV1/FVC、PEF),诱导痰中EOS计数,记录哮喘控制得分(ACT)及患者哮喘生命质量评分.哮喘组在治疗开始后1个月,2个月,3个月和6个月时进行重复测定.且以10名健康者做为对照组,同期测定上述观察指标.同时记录吸入用药后的不良反应.结果 研究共纳入38例患者,共有33例完成6个月或更长的随访观察.吸入糖皮质激素/长效β2受体激动剂复合制剂治疗后1个月,FEV1值明显改善[分别为(2.53±0.46)L,(2.89±0.62)L,P<0.01];3个月后上述变化更为显著达(3.19±0.47)L,与治疗1个月后相比发生显著变化(P<0.05);哮喘组诱导痰中EOS显著升高达(0.156±0.047)×10<'6>(P<0.05),激素吸入治疗过程中可见显著性变化,3个月降至(0.072±0.015)×10<'6>,6个月后痰中EOS计数明显减少,与治疗前相比较差异有统计学意义(q=6.58,P<0.05).吸入治疗后ACT评分明显改善,从治疗前(8±5)分增加至治疗后2个月(15±6)分,治疗后3个月到(20±4)分,与治疗前相比较差异有统计学意义(F=5.72,P<0.05).治疗6个月后,患者哮喘生命质量评分明显提高(P<0.05).哮喘组中共有12例患者(36.36%)获得完全控制.结论 联合吸入糖皮质激素/长效β2受体激动剂复合制剂治疗哮喘明显改善患者肺功能,减少痰EOS计数,有较好的临床疗效,且应至少连用6个月或以上.     

支气管哮喘儿童急性发作期诱导痰液T细胞亚群及自然杀伤T细胞的变化

目的 分析支气管哮喘(简称哮喘)儿童急性发作期及缓解期诱导痰液T细胞亚群及自然杀伤T细胞(NKT细胞)的变化,探讨T细胞及NKT细胞在儿童哮喘气道炎症中的作用.方法 选取18例哮喘急性发作、21例哮喘缓解期及12名正常儿童的痰液,采用流式细胞术比较各组儿童诱导痰液CD3、CD4、CD8、NKT(CD3~+ CD56~+)百分比及CD4/CD8比值.结果 哮喘急性发作组CD4细胞百分比[(43.75±13.5)%]明显高于缓解期组[(37.04±7.11)%]和正常对照组[(33.57±7.54)%](P＜0.05),CD8细胞百分比[(21.10±6.10)%]明显低于缓解期组[(28.67±5.32)%]和正常对照组[(28.31±9.46)%](P<0.05),CD4/CD8比值(2.14±0.94)高于缓解期组(1.33±0.35)和正常对照组(1.31±0.42)(P＜0.05),CD4、CD8细胞百分比及CD4/CD8缓解期组和正常对照组差异无统计学意义(P>0.05),急性发作组CD4/CD8比值与嗜酸粒细胞百分比呈正相关(r=0.559,P＜0.05).缓解期组CD4/CD8比值与嗜酸粒细胞无相关性(r=0.398,P>0.05).急性发作组CD3+ CD56+细胞百分比[(3.33±1.69)%]略高于缓解组[(3.09±1.23)%]及对照组[(2.94±0.87)%],但差异无统计学意义(P>0.05).CD3- CD56+细胞百分比在三组之间无统计学差异(P>0.05).结论 儿童哮喘急性发作期,气道内CD4细胞占优势,CD4/CD8细胞比例失衡,CD4细胞可能介导儿童哮喘气道炎症过程.     

LTC4及其合成酶与咳嗽变异性哮喘气道重建的相关性研究

目的研究白三烯C4（LTC4）及其合成酶表达与咳嗽变异性哮喘（CVA）气道重建的相关性,探讨CVA的发病机制,为CVA的早期诊断提供理论依据。方法将40例入选患者分为支气管哮喘组、CVA组各20例,另择20例健康人作为对照组。采用ELISA技术检测静脉血中LTC4的含量;应用RT-PCR检测LTC4合成酶在多形核细胞和淋巴细胞中的表达。选择对照组6例、支气管哮喘组6例、CVA组12例分别进行纤维支气管镜检查,取右中叶支气管组织,进行常规石蜡切片,HE染色,光镜下观察并测量基底膜厚度。结果支气管哮喘组LTC4含量及基底膜厚度显著高于CVA组和对照组,CVA组高于对照组（P〈0.01或〈0.05）。各组基底膜厚度与静脉血LTC4含量之间存在相关性（P〈0.05）。LTC4合成酶在CVA组和支气管哮喘组患者中的表达明显高于对照组（P〈0.05）。LTC4合成酶在多形核细胞中表达相对较强,在淋巴细胞中表达相对较弱。结论 CVA存在气道重建,静脉血中LTC4及其合成酶的表达可以作为CVA的快速诊断的指标之一。     

支气管哮喘患者不同时期诱导痰及血嗜酸粒细胞水平的观察

目的探讨诱导痰及末梢血嗜酸粒细胞(EOS)比例、诱导痰嗜酸粒细胞阳离子蛋白(ECP)水平在支气管哮喘（简称哮喘）发病不同时期的变化及应用价值。方法检测哮喘发作期40例（发作组）、哮喘缓解期48例（缓解组）、40名正常健康体检者（对照组）的诱导痰及末梢血EOS占白细胞百分比,同时分别测定痰ECP。结果 ①末梢血中EOS比...     

支气管哮喘患儿诱导痰嗜酸粒细胞与肺功能及FeNO浓度的关系

目的探讨支气管哮喘(哮喘)患儿诱导痰嗜酸粒细胞(EOS)与肺功能及呼出气一氧化氮(FeNO)浓度的关系。方法回顾分析了2016年10月至2018年12月在海南省妇女儿童医学中心儿科就诊的哮喘患儿共113例的病例资料,纳入观察组;将同期在海南省妇女儿童医学中心体检的50例健康儿童纳入对照组。分别检测2组受试者的诱导痰EOS、第1秒用力呼气容积占预计值百分比(FEV1%pred)以及FeNO水平并进行相关分析。结果观察组患儿诱导痰EOS和FeNO水平显著高于对照组(t=11.921、17.081,P值均<0.05),而其FEV1%pred较对照组差,差异有统计学意义(t=18.673,P<0.05)。针对观察组患儿的相关数据进行分析,发现FeNO与痰EOS水平呈正相关(r=0.265,P=0.005)。肺功能FEV1%pred与痰EOS水平无明显相关性(r=0.158,P=0.093)。结论哮喘患儿诱导痰EOS水平与FeNO呈正相关,但与肺功能无明显相关性。     

咳嗽变异型哮喘患者外周血细胞计数和血浆降钙素基因相关肽的水平观察

目的 探讨咳嗽变异型哮喘(CVA)患者外周血细胞计数、血浆降钙素基因相关肽(CGRP)的水平及其临床意义.方法 选择咳嗽变异型哮喘、典型支气管哮喘发作期患者各30例,对照组25名健康者,抽取清晨空腹静脉血5 ml,其中2 ml血行血常规检查,比较外周血中性粒细胞、嗜酸性粒细胞、淋巴细胞计数,3ml血用放射免疫法测定CGRP,对相关资料进行统计学分析.结果 (1)外周血细胞计数比较:中性粒细胞计数CVA组显著低于典型哮喘组与对照组(P＜0.05);嗜酸性粒细胞计数CVA组与典型哮喘组显著高于对照组(P＜0.05);淋巴细胞计数CVA组显著高于典型哮喘组与对照组(P＜0.05).(2)血浆CGRP水平比较:CVA组与典型哮喘组显著低于对照组(P＜0.01),CVA组与典型哮喘组比较差异无显著性(P＞0.05).结论 CVA患者外周血细胞计数是以嗜酸性粒细胞、淋巴细胞升高为主;CVA患者血浆CGRP水平显著低于对照组,提示CGRP在CVA发病过程中具有重要作用.