嗜酸粒细胞性支气管炎与咳嗽变异性哮喘患者的气道炎症特征的研究

目的 探究嗜酸粒细胞性支气管炎(EB)和咳嗽变异性哮喘(CVA)患者气道炎症细胞、细胞因子和炎性介质的特征,阐明两者存在不同气道炎性特征的可能机制.方法检测杭州市第一人民医院门诊收治的15例EB患者(EB组)、15例CVA患者(CVA组)、14例支气管哮喘(简称哮喘)患者(哮喘组)和14名健康体检者(健康对照组)诱导痰中嗜酸粒细胞(EOS)百分比;流式细胞仪检测白细胞介素(IL)-5及干扰素(IFN)-γ刺激的EOS表面CD69的表达;实时荧光定量PCR方法检测各组诱导痰上清液中前列腺素E2(PGE2)、白三烯C4(LTC4)、IL-5、IFN-γ mRNA的表达水平;酶联免疫吸附法(ELISA)检测各组诱导痰上清液中PGE2、LTC4、IFN-γ和IL-5蛋白表达水平.结果 EB组、CVA组、哮喘组诱导痰中EOS百分比分别为(15.8±3.2)%、(13.0±2.7)%和(11.6±4.5)%,均明显高于健康对照组的(1.0±0.4)%(均P<0.05).在IL-5和IFN-γ刺激下,EB组诱导痰中EOS表达CD69分别为1.49±0.42和1.51±0.52、CVA组分别为1.37±0.41和1.42±0.32、哮喘组分别为1.42±0.72和1.37±0.46,3组间差异无统计学意义,但较健康对照组(分别为0.42±0.21和0.39±0.12)差异均有统计学意义(均P<0.05).EB组、CVA组、哮喘组诱导痰中IL-5的mRNA及蛋白表达水平明显高于健康对照组(均P<0.05),但3组间差异无统计学意义;各组诱导痰中IFN-γ的mRNA及蛋白表达水平较健康对照组差异均无统计学意义.EB组诱导痰中PGE2浓度为(839±69)ng/L,明显高于CVA组的(33±8)ng/L、哮喘组的(25±6)ng/L和健康对照组的(24±8)ng/L(均P<0.01),后3组差异无统计学意义;EB组PGE2限速酶前列腺素氧化环化酶2(PTGS2)的mRNA水平表达量显著增加,较CVA组、哮喘组及健康对照组差异均有统计学意义(均P<0.01);CVA、EB和哮喘组诱导痰中LTC4浓度明显高于健康对照组(均P<0.05),CVA、EB及哮喘组中LTC4限速酶白三烯C4合成酶(LTC4S)的mRNA表达水平明显高于健康对照组(均P<0.05),EB组LTC4的mRNA及蛋白表达水平与CVA组和哮喘组比较,差异也有统计学意义(均P<0.05).CVA组、哮喘组诱导痰中LTC4/PGE2比值明显高于EB组(t值分别为8.67和13.12,均P<0.05).结论 EB患者诱导痰中PGE2高表达以及CVA组LTC4/PGE2比值较EB组显著增高,这两者可能是EB缺乏气道高反应性的炎症基础.

Abstract：

Objective To explore the characteristics of airway inflammatory cells, cytokines and inflammatory mediators in eosinophilic bronchitis (EB) and cough variant asthma (CVA) patients and to elucidate the underlying mechanism of distinct airway inflammation between EB and CVA. Methods This study included 15 patients with EB (EB group), 15 patients with cough variant asthma (CVA, CVA group), 14 patients with bronchial asthma (asthma group) and 14 healthy controls (healthy group). Percentage of eosinophils (EOS) in sputum induced by hypertonic saline was detected by FACS. The percentage of CD+69 EOS stimulated by interleukin-5 (IL-5) and interferon γ (IFN-γ) was also detected by FACS. The expression of leukotriene C4 synthase (LTC4S) and prostaglandin-endoperoxide synthase-2 (PTGS2) mRNA in sputum was measured by real-time PCR and the concentration of leukotriene C4 (LTC4) and prostaglandin E2(PGE2) in sputum was measured by ELISA. Results The percentage of EOS in induced sputum was 15.8±3.2 (EB group), 13.0±2.7 (CVA group) and 11.6±4.5 (asthma group), respectively, which were significantly higher than 1.0±0.4 in the healthy group. The difference was significant and the t value was 16.31, 15.23 and 14.21 respectively (P<0.05). After stimulated by IL-5 and IFN-γ, the percentage of CD+69 EOS in induced sputum was 1.5±0.4 and 1.5±0.5 (EB group), 1.4±0.4 and 1.4±0.3 (CVA group) and 1.42±0.72 and 1.37±0.46 (asthma group) respectively. There was no statistical significance between these 3 groups, but when compared with 0.4±0.2 and 0.4±0.1 in healthy group, the difference was significant(P<0.05). The expression of IL-5 mRNA and protein in induced sputum of EB group, CVA group and asthma group were higher than the healthy group and the difference was all statistically different (P<0.05), but there was no statistical significance between EB group, CVA group and asthma group. The expression of IFN-γ mRNA and protein in induced sputum of each group was not different when compared with healthy group (P＞0.05). The concentration of PGE2 in induced sputum of EB group was(839±69)ng/L, which was higher than (33±8) ng/L of CVA group, (25±6) ng/L of asthma group and (24±8) ng/L of healthy group (all P<0.01). There was no statistical difference between CVA group, asthma group and healthy group. The expression of PTGS2 in induced sputum of EB group increased significantly; when compared with CVA group, asthma group and healthy group, the difference was significant (all P<0.01). The concentration of LTC4 in induced sputum of EB group, CVA group and asthma group was all higher than the healthy group (all P<0.05). The expression of LTC4S mRNA of EB group, CVA group and asthma group was also higher than the healthy group (all P<0.05). The expression of LTC4S mRNA and LTC4 in the EB group was higher than that in the CVA group and the asthma group (P<0.05). The value of LTC4/PGE2 in the CVA group and the asthma group was higher than that in the EB group (t=8.7 and 13.1, P<0.05). Conclusion These data suggest that the difference in airway function observed in subjects with eosinophilic bronchitis and CVA (or asthma) may be due to the results of differences in PGE2 production and an imbalance between the production of bronchoconstrictor LTC4 and bronchoprotective PGE2 lipid mediators.     

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.     

慢性咳嗽患者咳嗽激发试验的意义

目的 初步观察咳嗽激发试验在不同性别、病程、病因的慢性咳嗽患者中的差异,以期为进一步评价咳嗽激发试验的临床意义及对临床诊疗慢性咳嗽提供启迪和帮助.方法 选择进行了咳嗽激发试验并最终确诊为上气道咳嗽综合征(UACS)、咳嗽变异性哮喘(CVA)、胃食管反流性咳嗽(GERC)的慢性咳嗽患者108例,分析咳嗽激发试验在不同病因、病程、性别的慢性咳嗽患者间的差异.结果 108例慢性咳嗽患者中UACS 76例,CVA 19例,GERC 13例.咳嗽敏感性阈值的对数(lgC5)女性患者(1.80)显著低于男性(2.40,P<0.05);病程≥12个月患者lgC5(1.80)显著低于病程<12个月的患者(2.40,P<0.05);不同病因患者lgC5存在显著差异,其中GERC患者的lgC5(1.49)显著低于CVA(2.40,P<0.05)和UACS(2.40,P<0.05)患者.结论 不同病因、病程、性别的慢性咳嗽患者咳嗽敏感性均存在差异,咳嗽激发试验在慢性咳嗽病因诊断中具有重要价值.

Abstract：

Objective Preliminary study of the cough reflex sensitivity test in chronic cough patients with different gender,disease duration and causes to evaluate the clinical diagnostic significance of the test and further provide some information for the treatment of chronic cough.Methods Totally 108 chronic cough patients in our hospital were enrolled in the study with the final diagnosis of upper airway cough syndrome(UACS),cough variant asthma(CVA)and gastroesophageal reflux cough(GERC).They all went through the cough reflex sensitivity test and the retrospective analysis of the results was made.Results In 108 patients with chronic cough(76 UACS,19 CVA and 13 GERC),lgC5 was significantly slower in the female(1.80)than the male(2.40,P<0.05)and in the patients with cough duration over 12 months(1.80)than those with cough duration under 12 months(2.40,P<0.05).Cough sensitivity in patients with different causes was significantly different(P<0.05)with lower lgC5 in GERC patients(1.49)than CVA (2.40,P<0.05)and UACS(2.40,P<0.05)patients.Conclusion Cough sensitivity is different in chronic cough patients with different gender,disease duration and causes.Cough reflex sensitivity test is of great value in the etiology diagnosis of chronic cough.     

室间隔缺损患儿封堵术前后血浆N末端B型利钠肽原和左心室Tei指数变化的意义

目的 探讨室间隔缺损(VSD)患儿封堵术前后血浆N末端B型利钠肽原(NT-proBNP)水平和左心室Tei指数变化的临床意义.方法选择膜固型VSD患儿60例作为VSD组,年龄相当的健康儿童30名作为正常对照组.采用电化学发光免疫学方法测定正常对照组与VSD组术前和术后5 min、24 h、1个月、3个月、6个月的血浆NT-proBNP水平.应用超声心动图测定正常对照组与VSD组术前和术后24 h、1个月、3个月、6个月左心室Tei指数.将VSD封堵术前后患儿的左心室Tei指数的变化与血浆NT-proBNP水平作相关性分析.结果 (1)VSD组术前血浆NT-proBNP水平显著高于正常对照组[(229.45±57.75)ng/L比(99.21±46.86)ng/L,P＜0.01];VSD组术后5 min、术后24 h血浆NT-proBNP水平均显著高于术前[(356.27±96.78)ng/L和(356.38±91.95)ng/L比(229.45±57.75)ng/L,均P＜0.01];VSD组术后1、3和6个月血浆NT-proBNP水平均显著低于术前[(131.33±34.79)ng/L、(96.56±31.55)ng/L和(93.39±29.46)ng/L比(229.45±57.75)ng/L,P＜0.05或P＜0.01];VSD组术后3和6个月与正常对照组比较差异无统计学意义(P＞0.05).(2)VSD组术前左心室Tei指数显著高于正常对照组(0.45±0.05比0.33±0.08,P＜0.01);VSD组术后24 h(0.52±0.05)与1个月(0.51±0.03)左心室Tei指数均显著高于术前和正常对照组(均P＜0.01);VSD组术后3和6个月左心室Tei指数均显著低于术前(0.34±0.07和0.34±0.06比0.45±0.05,均P＜0.01),与正常对照组比较差异无统计学意义(P＞0.05).(3)VSD患儿封堵前后左心室Tei指数的变化与血浆NT-proBNP水平呈正相关(r=0.653,P＜0.05).结论血浆NT-proBNP水平与左心室Tei指数可作为评价VSD患儿封堵前后心功能的参考指标.

Abstract：

Objective To explore the implication of the dynamic changes of plasma N-terminal proB-type natriuretic peptide (NT-proBNP) level and Tei index of left ventricle (LV) in children with ventricular septal defect(VSD) treated by transcatheter closure. Methods Sixty children with VSD treated by transcatheter closure with VSD occluder (Group VSD) and 30 healthy children (Group C) were included in this study. The plasma concentration of NT-proBNP, Tei index of LV and left ventricle ejection fraction (LVEF) were measured in Group C and at before, 5th minute, 4th hour, 1st month, 3rd month and 6th month after VSD closure in Group VSD. Results ( 1 ) The concentration of plasma NT-proBNP was significantly increased in children with VSD before transcatheter closure compared with Group C [(229.45 ±57.75 ) ng/L vs. (99. 21 ± 46. 86) ng/L,P ＜ 0. 01], significantly increased at 5th minute and 24th hour after transcatheter closure[( 356.27 ± 96. 78 ) ng/L and ( 356. 38 ± 91.95 ) ng/L vs. ( 229. 45 ± 57.75 ) ng/L, all P ＜0. 01], and significantly decreased at 1st month, 3rd months and 6th months after transcatheter closure [( 131.33 ± 34. 79 ) ng/L, (96. 56 ± 31.55 ) ng/L and ( 93. 39 ± 29. 46 ) ng/L vs. ( 229. 45 ± 57.75 ) ng/L,P＜0.05 or P＜0. 01]. (2) The Tei indexes of LV in Group VSD before transcatheter closure were significantly higher than in Group C (0. 45 ± 0. 05 vs. 0. 33 ± 0. 08, P ＜ 0. 01 ) and Tei index was significantly increased at 24th hour, 1 st month after transcatheter closure (P ＜ 0. 01 ) while significantly decreased at 3rd and 6th month compared with those before transcatheter closure (0. 34 ±0. 07 and 0. 34 ±±0. 06 vs. 0. 45 ±0. 05 ,all P ＜0. 01 ). (3) There is a positive correlation between the changes of the plasma concentration of NT-proBNP and the change of Tei index of LV before and after transcatheter closure (r = 0. 653, P ＜ 0. 05).Conclusion Tei index of LV and NT-proBNP can monitor cardiac function changes in children with VSD before and after transcatheter closure.     

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.     

不同病因儿童慢性咳嗽气道高反应的特征

目的 研究不同病因儿童慢性咳嗽的气道高反应性(AHR)特征.方法 纳入2012年4月至2013年12月于苏州大学附属儿童医院呼吸科就诊并明确为单病因的慢性咳嗽患儿,采用支气管激发试验测得FEV1下降20％时组织胺累计剂量(PD20),依据PD20将AHR严重程度分为重度、中度、轻度、极轻度4级,同时测定诱导痰嗜酸粒细胞(EOS)计数和呼出气一氧化氮(FeNO)浓度,按儿童慢性咳嗽指南推荐方案进行治疗,4周后随访,复查上述指标.结果 共纳入患儿66例,其中咳嗽变异性哮喘(CVA)17例,上气道咳嗽综合征(UACS) 37例,感染后咳嗽(PIC) 12例.CVA、UACS及PIC组AHR阳性初诊例数分别为17、13和9例,4周复诊时分别为17、5和2例.AHR严重程度分级,初诊时CVA、UACS和PIC组中～重度分别为5、0和0例,轻度～极轻度分别为12、13和9例,阴性分别为0、24和3例;4周后复诊CVA、UACS和PIC组中～重度度分别为1、0和0例,轻度～极轻度分别为16、5和2例,阴性分别为0、32和10例.CVA组PD20值为(0.47±0.28) mg,低于UACS组和PIC组[分别为(1.80±0.64)和(1.20±0.80) mg],差异均有统计学意义(均P＜0.01).CVA组的诱导痰EOS比例均＞3％,UACS和PIC组均＜3％.FeNO与痰EOS比例呈正相关(r=0.687,P=0.000).结论 不同病因儿童慢性咳嗽气道高反应的严重程度和持续时间不同,气道激发试验结合诱导痰对慢性咳嗽病因诊断有重要价值.     

Celecoxib inhibits proliferation and induces apoptosis via prostaglandin E2 pathway in human cholangiocarcinoma cell lines

AIM: To evaluate the roles and mechanisms of celecoxib in inducing proliferation inhibition and apoptosis of human cholangiocarcinoma cell lines. METHODS: Cyclooxygenase-2-overexpressing human cholangiocarcinoma cell line QBC939 and cyclooxygenase2-deficient human cholangiocarcinoma cell line SK-CHA-1were used in the present study. The anti-proliferative effect was measured by methabenzthiazuron (MTT) assay;apoptosis was determined by transferase-mediated dUTP nick end labeling (TUNEL) detection and transmission electron microscopy (TEN). Cell cycle was analyzed by flow cytometry (FCM). The PGE2 levels in the supernatant of cultured cholangiocarcinoma cells were quantitated by enzyme-linked immunoabsordent assay (ELISA). RESULTS: Celecoxib suppressed the production of PGE2and inhibited the growth of QBC939 cells. Celecoxib at 10,20, and 40 μmol/L inhibited PGE2 production by 26 %,58 %, and 74 % in QBC939 cells. The PGE2 level was much lower constitutively in SK-CHA-1 cells (18.6±3.2)compared with that in QBC939 (121.9±5.6) cells (P＜0.01)and celecoxib had no significant influence on PGE2 level in the SK-CHA-1 cells. The PGE2 concentration in SK-CHA-1cells also reduced but not significantly after treatment with celecoxib. The PGE2 concentration in SK-CHA-1 cells was (16.5±2.9) ng/well, (14.8±3.4) ng/well, (13.2±2.0) ng/well and (12.6±3.1) ng/well respectively, when pre-treated with 1 Jmol/L, 10 Jmol/L, 20 Jmol/L and 40 Jmol/L of celecoxib for 48 h (P＞0.05, vs control). The anti-proliferation effect of celecoxib (20 Jmol/L) on QBC939 cells was time-dependent,it was noticeable on day 2 (OD490=0.23±0.04) and became obvious on day 3 (OD490=0.31±0.07) to day 4 (OD490=0.25±0.06), and the OD490 in the control group (day 1)was 0.12±0.03 (P＜0.01, vscontrol). The anti-proliferation effect of celecoxib could be abolished by the addition of 200 pg/mL PGE2. The proliferation of SK-CHA-1 cells was inhibited slightly by celecoxib, the cell density OD490 in the presence of celecoxib and in control group was 0.31±0.04 and 0.42±0.03 respectively on day 2 (P＞0.05), 0.58±0.07 and 0.67±0.09 respectively on day 3 (P＞0.05), and 0.71±0.08 and 0.78±0.06 respectively on day 4 (P＞0.05). Celecoxib induced proliferation inhibition and apoptosis by G1-S cell cycle arrest: the percentage of QBC939 cells in G0-G1 phase after treatment with 40 Jmol/L (74.66±6.21) and 20 Jmol/L (68.63±4.36) celecoxib increased significantly compared with control cells (54.41±5.12, P＜0.01). The percentage of SK-CHA-1 cells in G0-G1 phase after treatment with various concentrations of celecoxib didn't change significantly compared with control cells. The TUNEL index was much higher in QBC939 cells treated with 20 Jmol/L celecoxib for 2 d (0.063±0.018) and for 4 d (0.102±0.037) compared with control cells (0.017±0.004, P＜0.01). CONCLUSION: The currentin vitro study indicates that inhibition of proliferation and induction of apoptosis in human cholangiocarcinoma cells by cyclooxygenase-2 specific inhibitor celecoxib may involve in COX-dependent mechanisms and PGE2 pathway. Celecoxib as a chemopreventive and chemotherapeutic agent might be effective primarily on COX2-expressing cholangiocarcinoma.     

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。     

依非韦仑为主的方案治疗人类免疫缺陷病毒/丙型肝炎病毒感染患者的安全性和有效性

目的 评价以依非韦仑为主的方案治疗HIV/HCV合并感染患者的安全性和有效性.方法 以依非韦仑为主的方案治疗53例HIV/HCV合并感染患者7年,观察CD4+T淋巴细胞计数、HIV RNA、肝功能、肝纤维化指标、血脂、血糖、血尿酸、血常规的变化.治疗前、后均数的比较采用t检验.结果 53例患者治疗前、后HIV RNA分别为(4.56±0.88)lg拷贝/mL和(1.70±1.10)lg拷贝/mL(t=14.781,P＜0.01);CD4+T淋巴细胞分别为(188.37±151.14)×106/L和(445.18±314.25)×106/L(t=5.362,P＜0.01);ALT分别为(36.6±16.3)和(57.2±9.9)U/L(t=7.864,P＜0.01);甘胆酸(CG)分别为(444.22±476.74)和(556.88±733.05)mg/L(t=0.938,P＜0.05);ⅣC分别为(45.13±8.25)和(47.88±4.51)ng/mL(t=2.129,P＜0.05);三酰甘油分别为(1.57±0.65)和(2.51±1.29)mmol/L(t=4.737,P＜0.01);血尿酸分别为(298.5±48.2)和(495.1±89.4)mmo1/L(t=14.092,P＜0.01).结论 以依非韦仑为主的方案治疗HIV/HCV合并感染患者是有效的,但可引起肝脏功能损害和代谢异常.

Abstract：

Objective To evaluate the efficacy and safety of efavirenz-based therapy in patients with human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infection. Methods Fiftythree HIV/HCV co-infected patients received efavirenz-based therapy were followed up for 7 years.The changes of CD4+ T lymphocyte count, HIV virus load, hepatic function, hepatic fibrosis index,blood lipid, blood glucose, blood uric acid and blood routine were observed. The comparison of means before and after treatment was performed by t-test. Results The HIV RNA levels at baseline and endpoint were (4. 56±0. 88) lg copy/mL and (1.70±1.10) lg copy/mL, respectively (t=14. 781, P＜0.01). The peripheral blood CD4+ T lymphocyte counts were ( 188.37±151.14)×106/L and (445.18±314.25)×106/L, respectively (t=5.362, P＜0.01).The alanine aminotransferase (ALT) levels were (36.6±16.3) U/L and (57.2±9.9) U/L, respectively (t=7.864, P＜0. 01).The glycocholic acid levels were (444.22±476.74) mg/L and (556.88±733.05) mg/L, respectively (t=0.938, P＜0.05). The Ⅳ-collagen(Ⅳ-C) levels were (45.13±8.25) ng/mL and (47.88±4.51) ng/mL, respectively (t= 2.129, P＜0.05). The riacylglycerol levels were (1.57±0.65)mmol/L and (2.51±1.29) mmol/L, respectively (t=4.737, P＜0.01). The blood uric acid levels were (298.5±48.2) mmol/L and (495.1±89.4) mmol/L, respectively (t= 14.092, P＜0.01).Conclusions The efavirenz-based therapy is efficacious in HIV/HCV co-infected patients, but it could cause liver injury and metabolic disorder.     

不同病因慢性咳嗽临床特征的诊断价值

目的 比较不同病因慢性咳嗽患者的特征及伴随症状,探讨其对临床慢性咳嗽的病因诊断的价值.方法 按慢性咳嗽病因诊断程序对2003年6月至2007年12月广州呼吸疾病研究所专科门诊的慢性咳嗽患者进行病因诊断.对单一病因的患者按常见病因进行分组,分析比较不同单一病因的慢性咳嗽患者的咳嗽时相、性质、诱发因素等特征及相关伴随症状.计量资料用x±s表示,计数资料以率表示,统计采用方差分析及卡方检验.结果 共入选196例单一病因的慢性咳嗽患者,男108例,女88例;年龄14～72岁,平均(40±12)岁.其中咳嗽变异性哮喘(CVA)50例,嗜酸粒细胞性支气管炎(EB)55例,上气道咳嗽综合征(UACS)45例、胃食管反流性咳嗽(GERC)46例.不同病因组慢性咳嗽患者年龄、性别及病程差异无统计学意义(均P>0.05).CVA患者以夜间为主的咳嗽发生率为26.0%(13/44),明显高于EB组[9.1%(5/55),x2=5.272,P<0.05]、UACS组[2.2%(1/45),x2=10.657,P<0.01]及GERC组[0%(0/46),x2=13.833,P<0.01],其诊断CVA的特异度为95.9%.GERC的进食相关咳嗽、反流相关症状的发生率分别为52.2%(24/46)和69.6%(32/46),均高于其他病因组(均P<0.05),诊断GERC的特异度分别为83.3%、80.0%.UACS的鼻后滴流感、鼻炎相关症状及鼻病史发生率为66.7%(30/45)、88.9%(40/45)及82.2%(37/45),均高于其他病因组(均P<0.05),诊断UACS的特异度分别为89.4%、65.6%及63.6%.结论 咳嗽的时相特征及部分伴随症状对单一病因慢性咳嗽的诊断具有一定的参考价值.     

老年人慢性咳嗽的病因构成和临床分析

目的 对老年慢性咳嗽患者采取经验性治疗联合病因诊断流程要求,分析老年人慢性咳嗽的病因及诊治效果. 方法 按照我国咳嗽指南,对52例老年慢性咳嗽患者,采取经验性治疗,效果不明确者,再采取病因诊断流程,包括肺功能、支气管舒张试验或支气管激发试验、诱导痰细胞学检查、鼻窦CT、24 h食管pH值监测等检查,进行病因判断和临床治疗. 结果 52例老年慢性咳嗽患者,明确病因76例次.老年人慢性咳嗽病因依次为咳嗽变异性哮喘(CVA)23例次(30.3％),上气道咳嗽综合征(UACS) 16例次(21.1％),胃食管反流性咳嗽(GERC)12例次(15.8％),药源性咳嗽(DC)9例次(11.8％),脑血管疾病7例次(9.2％),肺间质纤维化、嗜酸粒细胞性支气管炎(EB)、变应性咳嗽(AC)、左心功能不全各1例次(各占1.3％),病因未明4例次(5.3％).单一病因31例(59.6％),二重病因11例(21.2％)、三重病因5例(9.6％)、四重病因1例(1.9％). 结论 CVA、UACS、GERC和DC是最常见的老年人慢性咳嗽的病因,遵循我国咳嗽指南,根据常见病因采取经验性治疗与病因诊断相结合,可较好适合我国基层医院的诊疗需求.     

"不明原因"慢性咳嗽的病因构成和临床特征分析

目的 分析经遵循我国慢性咳嗽诊断流程明确的慢性咳嗽患者的病因构成和临床特征,为经验性诊治慢性咳嗽提供依据.方法 对"不明原因"慢性咳嗽患者,遵循我国咳嗽指南慢性咳嗽诊断流程,在询问病史和查体的基础上,进行常规肺通气功能测定和支气管激发试验、诱导痰细胞学检查、鼻窦X线片或CT、24 h食管pH值监测等检查,根据检查结果和治疗后反应,最后确定病因诊断.收集卫生部中日友好医院患者病因资料并与广州呼吸疾病研究所的研究结果进行比较,以x2检验比较构成比差异.结果 2005年10月至2009年2月共收集不明原因慢性咳嗽患者103例,明确诊断咳嗽病因123例次.慢性咳嗽病因分布依次为咳嗽变异性哮喘41例次(33.3%),鼻炎/鼻窦炎30例次(24.4%),胃食管反流性咳嗽25例次(20.3%),药物性咳嗽7例次(5.7%),嗜酸粒细胞性支气管炎6例次(4.9%),变应性咳嗽4例次(3.3%),肺间质纤维化2例次(1.6%),左心功能不全和支气管扩张各1例次(0.8%),病因未明6例次(4.9%).单一病因咳嗽者86例(83.5%),二重病因14例(13.6%)、三重病因3例(2.9%).咳嗽变异性哮喘患者夜间咳嗽发生率最高[80.9%(36/41),x2=19.81,P<0.01],63.4%(26/41)合并鼻炎,68.3%(28/41)有季节性加重,其中67.8%(19/28)于秋季加重;胃食管反流性咳嗽以日间咳嗽为主,56.0%(14/25)有进食相关性咳嗽,68.0%(17/25)伴有反流相关症状;鼻炎/鼻窦炎患者有痰咳嗽发生率最高[73.3%(22/30),x2=24.99,P<0.01].与广州资料相比,咳嗽变异性哮喘和胃食管反流性咳嗽比例高于广州(x2值分别为9.52和4.56,均P<0.01),而嗜酸粒细胞性支气管炎和变应性咳嗽比例低于广州(x2值分别为17.61和7.86,均P<0.01).结论 慢性咳嗽常见病因构成为咳嗽变异性哮喘、鼻炎/鼻窦炎、胃食管反流性咳嗽、药物性咳嗽、嗜酸粒细胞性支气管炎和变应性咳嗽,与广州资料比较具有一定的差异.各种病因的咳嗽具有一定的临床特征,慢性咳嗽的病因构成和临床特征可为经验性诊治慢性咳嗽提供依据.     

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。     

呼出气一氧化氮在慢性咳嗽严重程度评估中的应用价值

目的 探讨呼出气一氧化氮(FeNO)在慢性咳嗽严重程度评估中的临床价值.方法 根据我院呼吸内科门诊2014年6月至2015年2月明确诊断为慢性咳嗽的连续病例68例,对所有患者根据FeNO水平分组,研究组间主观评分差异,并在治疗4周后进行FeNO检测、并评价咳嗽症状积分、莱赛斯特生活质量问卷(LCQ)评分.结果 以FeNO水平分组后,患者LCQ评分及咳嗽症状日间积分各组间差异有统计学意义(P＜0.01).将患者根据咳嗽病因分组,咳嗽变异性哮喘(CVA)组FeNO均值[(67.5±17.1) ppb]显著高于EB组[(34.6±9)ppb],2组高于其他病因组[(23.4±8.9) ppb](Z=-4.9,P＜0.01),治疗后FeNO水平、LCQ值和咳嗽积分相较治疗前均有改善(Z=-3.975,P＜0.01),3组病因间FeNO下降程度不同(P＜0.01),CVA组平均改善率[(33.2±3.9)％]高于其他2组病因患者,FeNO治疗前后总体下降变化与LCQ评分(r=0.48,P＜0.01)及咳嗽症状积分变化呈正相关,其中CVA组及EB组与LCQ评分治疗变化呈较强的相关性,相关系数分别为0.55和0.66(P＜0.01).结论 FeNO与患者的咳嗽症状和生活质量存在相关性,在慢性咳嗽病情评估中具有重要参考价值.     

胃食管反流性咳嗽患者气道黏膜与分泌物中神经肽含量的变化

目的探讨气道神经源性炎症在胃食管反流性咳嗽(GERC)发病中的作用。方法20例GERC患者(GERC组),与10名正常人(正常对照组)和8例其他病因咳嗽伴有胃食管反流(GERD)患者(GERD组)分别作对比分析,观察GERC患者治疗前、后气道黏膜与分泌物中神经肽的变化。用酶联免疫吸附测定(ELISA)法测定痰上清液P物质(SP)、神经肽A(NKA)、神经肽B(NKB)的含量;用放射免疫法测定痰上清液降钙素基因相关肽(CGRP)的浓度。用免疫组化法检测气道黏膜、诱导痰细胞SP、NKA及SP受体(NK1)的表达,结果用半定量病理学方法表示。结果GERC组患者痰上清液SP、CGRP含量[(266±207)ng/L、(180±83)ng/L]显著高于正常对照组[(143±36)ng/L、(105±64)ng/L,P分别<0.05、<0.01]和GERD组[(130±11)ng/L、(89±16)ng/L,P分别<0.05、<0.01],NKA、NKB的含量各组间比较差异无统计学意义(P均>0.05);GERC组痰细胞SP、NK1和NKA的表达显著高于正常对照组(P分别<0.01、<0.05,<0.05)和GERD组(P均<0.05);GERC组气道黏膜SP的表达也显著高于GERD组(P<0.01)。治疗后,诱导痰细胞SP、NK1、NKA的表达较治疗前显著降低(P<0.01,<0.01或<0.05),痰上清液CGRP的含量也较治疗前显著降低(P<0.05)。结论GERC患者气道存在明显的神经源性炎症,治疗后神经肽趋于正常,提示神经源性炎症与GERC的发病密切相关。