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可接受的诱导痰标本可以在健康和哮喘儿童使用甘露醇激发试验中获得GydF4y2Bahttp://ow.ly/KSd2MGydF4y2Ba
致编辑:GydF4y2Ba
中号annitol is used as an indirect challenge test to assess airway hyperresponsiveness in asthmatic adults and children as young as 4 years old [1GydF4y2Ba-GydF4y2Ba4GydF4y2Ba]。最近,它已成功在成人痰诱导试验[GydF4y2Ba五GydF4y2Ba,GydF4y2Ba6GydF4y2Ba]。在儿童中,诱导痰通常是由高渗盐溶液(HSS)获得的并且具有几个适应症,如在感染性疾病样品中获得[GydF4y2Ba7GydF4y2Ba],监测重度哮喘[GydF4y2Ba8GydF4y2Ba]并限定哮喘表型[GydF4y2Ba9GydF4y2Ba]。GydF4y2Ba
据我们所知,一直没有关于使用甘露醇获得儿童诱导痰研究。这项研究的目的是评估甘露醇激发试验的可行性和安全性,以获得在幼童诱导痰。这项初步研究是一个项目,研究哮喘和非哮喘儿童滑雪者支气管反应和炎症的一部分。GydF4y2Ba
这是一个横截面,观察性和描述性研究。Healthy and asthmatic children (7–13 years old) were randomly selected from a ski club (Ordino Arcalís Ski Club, Ordino Arcalís, Andorra). The study was approved by the Ethics Committee of Nostra Senyora de Meritxell Hospital (Escaldes-Engordany, Andorra), and written informed consent from parents as well as authorisations from the ski club were obtained for participation in the study. Inclusion criteria for asthmatic children were diagnosis of bronchial asthma, regardless of the severity, and absence of asthma symptoms with no need for short-acting β2GydF4y2Ba上月激动剂或口服皮质类固醇。GydF4y2Ba
该协议研究包括:带有NIOX MINO设备(Aerocrine,索尔纳,瑞典)呼出一氧化氮分数的选择;皮肤点刺试验(ALK-ABELLO过敏原提取; ALK,圆石,TX,USA)为常见的呼吸道过敏;和支气管激发试验和痰诱导吸入甘露醇。GydF4y2Ba
甘露醇挑战试验通过渐进协议来执行[GydF4y2Ba2GydF4y2Ba]。dry powder mannitol (Osmohale; Aldo-Unión, Esplugues de Llobregat, Spain) was inhaled in step-wise doses (0, 5, 10, 20, 40, 80, 160, 160, 160 mg). Measurements of forced expiratory volume in 1 s (FEV1GydF4y2Ba) were taken pre-dose and 1 min post-dose, with the highest value for FEV1GydF4y2Ba被记录。阳性反应被认为是当FEVGydF4y2Ba1GydF4y2Ba下降≥15%或当FEVGydF4y2Ba1GydF4y2Ba下跌连续剂量之间≥10%。GydF4y2Ba
为了获得诱导痰,孩子鼓励每个甘露醇剂量的吸入后向痰咳出到无菌容器中。GydF4y2Ba
如先前所述进行痰液检查[GydF4y2Ba10GydF4y2Ba],以下P的方法GydF4y2BaizzichiniGydF4y2Ba等GydF4y2Ba。[GydF4y2Ba11GydF4y2Ba]。简言之,痰污染唾液和称重分离。Samples had to weigh ≥50 mg to be accepted as valid. The aliquot was then dispersed with four volumes of freshly prepared dithiothreitol solution. The mixture was vortexed, and after being rocked for 15 min, four volumes of Dulbecco's PBS were added. The suspension was then filtered through 48-mm nylon gauze (Millipore Corporate Headquarters, Billerica, MA, USA) and centrifuged at 314×GGydF4y2Bafor 10 min. This resulted in the formation of a cell pellet and supernatant solution. The supernatant was stored at −80°C for analysis of inflammatory factors. The proportion of salivary squamous cells was noted and cell viability was determined using the trypan blue exclusion method. The specimens were considered adequate if squamous cells represented <20% and viable cells ≥60% of the total cell population. Cytospin cell preparations were made using a cytocentrifuge. The cytospin slides were stained with May–Grünwald–Giemsa to determine the differential cell count, made by counting 400 cells per slide. Sputum eosinophilia was established when the percentage of eosinophils was ≥2.5% [12GydF4y2Ba]。GydF4y2Ba
我们研究了7个哮喘和10名健康儿童(GydF4y2Ba表格1GydF4y2Ba)。所有,但哮喘的一个孩子被认为是特应性。3例患者有间歇性发作性哮喘和四有持续性哮喘,以及吸入类固醇的两名患者,并与奥马珠单抗在其他两个病人控制。GydF4y2Ba
在17名儿童(76.5%)的13,得到一个成功的诱导痰,允许细胞结构痰的充分研究(GydF4y2Ba表格1GydF4y2Ba)。我们没有在三个健康和一个哮喘孩子获得良好的痰标本:两个健康的男孩的53%和54%,一个健康的女孩有0%生存能力和一个哮喘男孩,有66%生存力的,但25%的鳞状一个可行性细胞的污染。GydF4y2Ba
激发试验耐受性好,不良反应在所有患者无需停止在任何情况下测试温和;咳嗽(N = 17),吞气症(N = 3),呕吐(N = 2),咽喉痛(N = 7),在嘴不好的味道(N = 10)。甘露醇试验在所有的孩子时间中位数为55(范围50-59)分钟。GydF4y2Ba
In this preliminary study, we report that sputum can be successfully induced in children aged 7–13 years with inhaled mannitol. The success rate of this technique to obtain a good sputum sample was somewhat higher than our previous results using HSS: 62% in 108 children [10GydF4y2Ba]。GydF4y2Ba
在先前的研究成人,由受试者89%甘露糖醇的挑战期间盐水挑战和受试者76%期间获得诱导痰。有一个较低的总细胞数甘露醇GydF4y2Ba与GydF4y2Ba高渗盐水诱导痰。然而,痰嗜酸粒细胞和中性粒细胞没有显著不同,并使用甘露醇,受试者比例较高被分类为嗜酸性粒细胞具有哮喘[GydF4y2Ba五GydF4y2Ba]。GydF4y2Ba
在我们的研究中,最常见的不良事件是咳嗽,所有的孩子比较只有29%在我们与HSS [先前公布的研究哮喘患儿的试验过程中咳嗽GydF4y2Ba10GydF4y2Ba]。然而,这是的甘露醇的效果,这可能是有用的,以诱导痰液的固有特性和咳嗽并不重要足够以停止测试。GydF4y2Ba
否则,支气管激发试验在七个哮喘的四个孩子和三个10名健康儿童为阳性。需要进一步研究有关在儿童滑雪者的特异性和测试的灵敏度。GydF4y2Ba
综上所述,尽管小样本,在获得高质量的痰标本的成功率相当高。根据我们的研究结果,甘露醇激发试验可在年幼的孩子使用,同时为了研究这两个高反应性和炎症进行支气管激发试验和诱导痰。GydF4y2Ba
脚注GydF4y2Ba
支持声明:这项研究是由阿尔工会(埃斯普卢加斯,西班牙)的支持。GydF4y2Ba
利益冲突:无申报。GydF4y2Ba
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- 公认GydF4y2Ba2015年3月15日。GydF4y2Ba
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