Plasma membrane polarity of polymorphonuclear leucocytes from children with primary ciliary dyskinesia

BACKGROUND: Polymorphonuclear leucocytes (PMN) from subjects with primary ciliary dyskinesia (PCD) can have abnormal locomotory systems. The locomotory activity of PMN is the result of biochemical events mediated by the plasma membrane. In this study we investigated plasma membrane polarity of PMN from children with PCD. DESIGN: Membrane polarity was studied in 11 children with PCD and in healthy controls by measuring the steady-state fluorescence excitation and emission spectra of 2-dimethylamino[6-lauroyl]naphthalene (Laurdan), which is known to be incorporated at the hydrophobic-hydrophilic interface of the bilayer, displaying spectral sensitivity to the polarity of its surroundings. Laurdan shows a marked steady-state emission red shift in polar solvents, with respect to nonpolar solvents. Moreover, the effect of the microtubule disassembling agent colchicine on PMN membrane polarity was evaluated. RESULT: Our results show a red shift of the fluorescence excitation and emission spectra of Laurdan in PMN from the PCD group with respect to the control group. These data indicate an increase in membrane polarity of PMN from the PCD group. Treatment of PMN with colchicine induced a red shift in the Laurdan excitation and emission spectra with the same trend observed in PMN from the PCD group. CONCLUSION: PMN from children with PCD are characterized by an increased plasma membrane polarity. These changes could be the basis of the modifications in the locomotory activities of PMN. The observed alterations may be attributed to abnormalities in the cytoskeleton.     

Situs inversus totalis and prenatal diagnosis of a primary ciliary dyskinesia

Situs inversus, a condition in which the major visceral organs are reversed from their normal positions in the body, can be detected by prenatal ultrasonography. Often benign, it may be associated with primary ciliary dyskinesia, an autosomal recessive disorder characterized by chronic respiratory disease. Yet, prenatal diagnosis of primary ciliary dyskinesia has not been reported. We describe a pregnancy in which situs inversus was diagnosed by fetal ultrasound at 20 weeks gestation. Prenatal testing for primary ciliary dyskinesia led to the discovery that both parents were asymptomatic carriers of a pathogenic mutation in the CCDC103 gene, with an affected neonate.     

Repeatability of exhaled nitric oxide measurements in patients with COPD

The assessment of the presence of eosinophilic airway inflammation may help in predicting the steroid response in subjects with respiratory symptoms. Unlike patients with asthma, only a subset of patients with chronic obstructive pulmonary disease (COPD) benefits from steroid treatment. Fractional exhaled nitric oxide (FENO) is a useful surrogate marker for eosinophilic airway inflammation, but data on the repeatability of FENO measurements in COPD needed for the assessment of significant change are insufficient. The aim of this study was to assess the short-term repeatability of FENO measurement in subjects with moderate to very severe chronic airway obstruction compared to that in healthy subjects. We studied 20 patients with stable COPD and 20 healthy subjects, and determined FENO (flow rate 50 ml s(-1) ) three times: at baseline, 10 min and 24 h after baseline. Spirometry was performed on the first study day after the FENO measurements. The median FENO concentration in patients with COPD was 15·6 ppb, and in healthy subjects, 15·2 ppb. The coefficient of variation (CoV) for 24-h measurements was 12·4% in COPD patients, and 15·9% in healthy subjects. Among COPD patients with global initiative for chronic obstructive lung disease stage 2 disease, the CoV was 13·7%, and among those with stage 3-4 disease, 10·5%. The findings indicate that the short-term repeatability of FENO measurement in patients with moderate to very severe COPD is equally good as in healthy subjects. A change in FENO exceeding 24% is likely to reflect a minimum measurable change in COPD.     

呼出气一氧化氮对呼吸机相关性肺炎的早期诊断意义

目的:探讨呼出气一氧化氮(FeNO)对呼吸机相关性肺炎(VAP)的早期诊断意义。方法:选择入住首都医科大学附属复兴医院重症医学科(ICU)需要进行有创机械通气的患者。收集患者人口学资料,第1、3、5、7天FeNO、白细胞计数(WBC)、降钙素原(PCT)及预后指标,前瞻性观察FeNO是否对VAP具有早期诊断意义。按照入ICU的主要原因分为肺内炎症组、肺外炎症组及非炎症组;将机械通气时间≥3 d的患者,根据14 d内是否发生VAP分为VAP组和非VAP组。结果:肺内炎症组患者FeNO浓度明显高于肺外炎症组及非炎症组(P0.05)。与非VAP组患者相比,VAP组患者第3天和第5天的FeNO明显升高,其对VAP的发生与否有良好的临床预测价值(第3天:AUC 0.87,P0.001,分界点6.5 ppb,敏感性76.9%,特异性81.4%;第5天:AUC 0.75,P=0.001,分界点为5.5 ppb,敏感性73.1%,特异性67.4%)。与非VAP组患者相比,VAP组患者28 d内非机械通气时间缩短(P0.05)、ICU住院时间延长(P0.05)。结论:肺炎患者FeNO明显升高,升高的FeNO对VAP有很好的临床预测价值,在临床中可以作为VAP的一项生物标志物。     

Validation study of nasal nitric oxide measurements using a hand-held electrochemical analyser

Background Exhaled nitric oxide (NO) measurement is a simple and non‐invasive method for monitoring airway inflammation. Similarly, nasal NO has been proposed as a surrogate marker in inflammatory diseases of the upper airways, e.g. allergic rhinitis. A new portable analyser using an electrochemical sensor has been developed for measurements of exhaled NO, and its reproducibility and comparison with other analysers has been tested recently in healthy subjects and in patients with lower airways disease. The application of this hand‐held analyser in nasal NO analysis was tested and compared to the gold standard represented by a chemiluminescence analyser. Materials and methods Thirty subjects including 15 patients with allergic rhinitis (AR) and 15 healthy subjects (HS) were studied. The intraindividual variability, calculated as the difference in nasal NO levels between two measurements from a single nasally exhaled breath manoeuvre, and the comparison between the electrochemical analyser (NIOX MINO, Aerocrine) and a chemiluminescence analyser (NOA, Sievers) were performed. Results In AR patients mean nasal NO was 59·0 ± 16·3 p.p.b. with the MINO and 58·3 ± 15·6 p.p.b. with the NOA. In HS nasal NO was 49·1 ± 10·8 p.p.b. with the MINO and 49·8 ± 8·2 p.p.b. with the NOA. The Bland‐Altman analysis showed bias values of 0·005 ± 3·6 with the 95% limits of agreement from –6·97 to 6·98 p.p.b. Conclusion Measurements of nasal NO levels with a hand‐held electrochemical analyser are reproducible and the results are comparable to a stationary chemiluminescence analyser.     

Repeatability of successive measurements with a portable nitric oxide analyser in patients with suggested or diagnosed asthma

The objective of our study was to determine the repeatability of successive measurements of exhaled nitric oxide (NO) with a portable device (NIOX MINO^®). We assembled a sample of 332 pairs of repeated NO measurements acquired in the same session. The mean of the two successive measurements varied from 4.0 to 279.5 parts per billion (ppb). The mean coefficient of repeatability (CoR) of the entire study sample was 2.4?ppb. The difference between paired measurements increased slightly with increasing fractional exhaled nitric oxide (FENO); for the mean FENO of 24.0?ppb the calculated CoR was 2.2 and for the FENO of 54?ppb it was 3.2?ppb. The difference between paired observations for FENO values <50?ppb was significantly lower than that for values >50?ppb (p<0.001). The demonstrated repeatability of successive measurements with the device supports the view that, when the exhalation manoeuvre is technically valid and FENO is within normal limits, repeated determinations of FENO are generally not necessary for screening. In other cases, the mean value of two successive measurements would represent a more reliable measurement result.     

Nasal nitric oxide in cystic fibrosis with and without humming

BACKGROUND: Nasal nitric oxide (nNO) values are reduced in patients with cystic fibrosis (CF). Humming during nNO measurement increases nNO values in healthy subjects. Nasal NO is reduced in patients with CF, sinus disease or nasal polyps. Humming nNO values have not been reported in CF patients yet. Our aim was to explore humming nNO values in CF patients and assess whether nNO during humming is a better discriminator than silent nNO measurements in this patient group. MATERIALS AND METHODS: In a cross sectional study we measured nNO concentrations in healthy controls (HC) and in CF patients (n = 23 and 31, respectively). The participants held their breath for 10 s while air was passively extracted from one nostril with 700 mL min(-1) for direct NO measurements (NIOX chemiluminescence analyser). Subsequently nNO was measured during humming with the mouth closed for 10 s. RESULTS: Mean nNO in parts per billion (p.p.b.) (SD) during breath hold was 499 (164) and 240 (139), respectively. The median nNO peak (p.p.b., minimum-maximum) during humming was 1500 (425-4100) for HC and 120 (23-500) for CF. There was a highly significant difference between nNO both with and without humming between CF and HC (P < 0.01). The sensitivity and specificity of nNO for detecting CF were better with humming. CONCLUSION: Nasal NO concentrations with and without humming are significantly decreased in CF. Humming nNO is an excellent discriminator between HC and CF and performs better than silent nNO.     

肺泡气一氧化氮在初始治疗前低FeNO哮喘患者治疗中的价值

目的 探讨初始治疗前低呼出气一氧化氮(FeNO)哮喘患者中,肺泡气一氧化氮(CaNO)水平升高能否作为吸入糖皮质激素/长效β2受体激动剂(ICS+LABA)治疗获益的预测因子.方法 收集该院呼吸与危重症医学科就诊的具有哮喘临床特征、舒张试验阳性且FeNO≤50 ppb的患者60例作为研究对象,依据CaNO水平是否升高分...     

呼出气一氧化氮对哮喘的诊断作用及与过敏原sIgE的关系

目的探讨呼出气一氧化氮(FeNO)对哮喘的诊断作用及与过敏原特异性IgE抗体(sIgE)的关系。方法选取2017年8月至2019年8月收治的98例疑似哮喘患儿进行观察,收集所有患儿的临床特征指标,检测肺功能及FeNO浓度,分析FeNO对哮喘的诊断作用及其与过敏原sIgE的关系。结果哮喘组的FeNO水平高于非哮喘组(P<0.05)。血清过敏原sIgE阳性患儿的Fe NO水平高于阴性患儿(P<0.05)。经Pearson相关性分析得出,FeNO水平与血清过敏原sIgE、血清总IgE和血清过敏原种类呈显著正相关(r=0.703、0.624、0.719,P<0.05)。结论FeNO在哮喘中具有一定的诊断价值,与过敏原sIgE存在显著的相关性,可为临床诊断与治疗哮喘提供有利参考依据。     

The accuracy of primary care teams in diagnosing disorders of the shoulder

Objective The aim of this retrospective, observational study was to determine the accuracy of diagnoses given by a multitude of primary care services to patients referred to an orthopaedic upper limb surgeon with a focus towards shoulder pathology. Methods Records of all patients referred to a single upper limb surgeon over a 1‐year period were reviewed. The diagnosis in the primary care setting was compared with the initial working diagnosis in the specialist clinic and against the final diagnosis following specialist investigation/intervention. Results 114 eligible patients were identified. General practitioners referred 35% of patients, musculoskeletal triage services referred 63% of patients and independent physiotherapists referred 2% of patients. It was found that 37% of patients were not given a diagnosis by the primary care team. When a diagnosis was given, accuracy was 50% (κ = 0.28) when correlated against that given by an orthopaedic upper limb surgeon. Conclusions Our findings suggest that knowledge of shoulder conditions is limited amongst primary care practitioners. There does not appear to be any difference in accuracy between general practitioners and musculoskeletal triage services.     

Measuring diagnostic and predictive accuracy in disease management: an introduction to receiver operating characteristic (ROC) analysis

Diagnostic or predictive accuracy concerns are common in all phases of a disease management (DM) programme, and ultimately play an influential role in the assessment of programme effectiveness. Areas, such as the identification of diseased patients, predictive modelling of future health status and costs and risk stratification, are just a few of the domains in which assessment of accuracy is beneficial, if not critical. The most commonly used analytical model for this purpose is the standard 2 x 2 table method in which sensitivity and specificity are calculated. However, there are several limitations to this approach, including the reliance on a single defined criterion or cut-off for determining a true-positive result, use of non-standardized measurement instruments and sensitivity to outcome prevalence. This paper introduces the receiver operator characteristic (ROC) analysis as a more appropriate and useful technique for assessing diagnostic and predictive accuracy in DM. Its advantages include; testing accuracy across the entire range of scores and thereby not requiring a predetermined cut-off point, easily examined visual and statistical comparisons across tests or scores, and independence from outcome prevalence. Therefore the implementation of ROC as an evaluation tool should be strongly considered in the various phases of a DM programme.     

Exogenous nitric oxide regulates activity and synthesis of vascular endothelial nitric oxide synthase

Background Nitric oxide (NO) – a major signalling molecule of the vascular system – is constitutively produced in endothelial cells (EC) by the endothelial NO synthase (eNOS). Since a reduced NO synthesis is an early sign of endothelial dysfunction and NO delivering drugs are used to substitute the impaired endothelial NO production, we addressed the effect of exogenous NO on eNOS in human umbilical venous endothelial cell cultures. Materials and methods The synthetic NO donor DETA/NO (trade name, but in the following we refer to detNO), that releases NO in a strictly first order reaction with a half life of 20 h, was used in our experiments. Results Short‐term (20–30 min) detNO treatment of EC increases the Ser¹¹⁷⁷ phosphorylation of the constitutively expressed endothelial NOS and the production of endogenous NO generated by eNOS from [³H]arginine. The phosphorylation of eNOS is Akt‐dependent and completely reverted by the phosphatidylinositol‐3 kinase (PI‐3K) inhibitor LY294002. A prolonged continuous exposure of EC to detNO 150 µmol L^?1 over a period of 24–48 h causes a reversible cell cycle arrest at G₁‐phase associated with a larger cell volume and increased cell protein content (hypertrophic phenotype of EC). The eNOS protein and mRNA of the hypertrophic cells and the generation of endogenous NO are reduced but eNOS phosphorylation could still be elevated by stimulation with vascular endothelial growth factor. Conclusions Our data explain clinical studies describing a short‐term but not a long‐term benefit of NO treatment for patients with cardiovascular risk factors. The results could be a rational approach to develop a generation of NO donors accomplishing a retarded release from NO donors that mimic the low continuous pulsatile stress‐induced release of endogenous NO.     

原发性肝癌血管内皮生长因子及一氧化氮的检测及意义

目的 探讨原发性肝癌患者治疗前后血清血管内皮生长因子(VEGF)及一氧化氮(NO)的变化及临床意义.方法 采用酶联免疫吸附试验(ELISA),分光光度法对35例原发性肝癌患者,30例健康人的血清进行检测.结果 肝癌患者治疗前血清VEGF 362.39±72.42 pg/ml,血清NO 94.23±21.24 μmol/L,高于健康对照组(P＜0.01);肝癌稳定组治疗前后血清VEGF分别为324.51±44.24和128.93±40.86 pg/ml,血清NO分别为82.42±4.80和75.76±10.01 μmol/L,VEGF在治疗前后有统计学意义(P＜0.01),NO在治疗前后差异无统计学意义(P＞0.05);肝癌恶化组治疗前后血清VEGF分别为384.77±77.20和428.77±87.29 pg/ml,差异有统计学意义(P＜0.01),血清NO分别为101.21±24.08和119.32±20.53μmol/L差异有统计学意义(P＜0.01).结论 血清VEGF及NO的检测可以为临床原发性肝癌的治疗及预后判定提供依据.     

呼出气一氧化氮联合相关因素对咳嗽变异性哮喘的诊断价值评价

目的 :评价呼出气一氧化氮(Fe NO)检测单独或联合咳嗽变异性哮喘(cough variant asthma,CVA)相关因素对诊断CVA的有效性和准确性。方法:连续纳入2010年8月至2011年10月本院呼吸科门诊因慢性咳嗽行支气管激发试验检查的患者297例,记录病史并测定呼出气Fe NO水平,以支气管激发试验阳性作为CVA诊断的金标准,绘制ROC曲线,探讨诊断CVA的Fe NO临界点。建立Logisitic回归模型,通过分析筛选与CVA相关的因素,根据ROC曲线评价Fe NO联合其他相关因素对CVA的诊断价值。结果:145例患者支气管激发试验阳性并排除其他疾病被诊断为CVA(哮喘组),152例支气管激发试验阴性患者诊断为非哮喘的慢性咳嗽(非哮喘组)。1哮喘组患者Fe NO水平明显高于非哮喘组[(45.33±38.86)ppb比(26.28±23.86)ppb,P     

呼出气一氧化氮测定在支气管哮喘诊断和管理中的价值

慢性气道炎症是支气管哮喘的本质。临床上急需能客观准确地反映气道炎症的炎性标志物来指导哮喘的诊断和管理,呼出气一氧化氮(FeNO)测定是一种符合临床需要的快速检测方法,现就FeNO在哮喘诊断、气道炎症监测、哮喘严重程度评级及预测哮喘发作等方面的研究进展做一综述。     

呼出气一氧化氮检测在哮喘儿童孟鲁司特治疗中的应用价值

目的探讨呼出气一氧化氮(FeNO)检测在哮喘儿童孟鲁司特治疗中的应用价值。 方法选择2013年5月至2014年5月于复旦大学附属上海市第五人民医院儿科门诊就诊的5～14岁哮喘儿童,共40例。给予入组患儿孟鲁司特常规治疗12周,随访6次并检测入组患儿FeNO水平,测定晨起呼气峰流速(PEF),并计算PEF占预计值的百分比(PEF%pred),记录儿童哮喘控制评分(C-ACT)。根据孟鲁司特治疗前及治疗12周时的哮喘控制水平分级变化,将入组患儿分为显效组和无效组,采用SPSS 17.0对不同分组的临床资料进行分析。最后有31例患儿完成为期12周的研究,其中显效组21例,无效组10例。 结果两组患儿在哮喘病史时间[(2.36±2.03)年,(3.60±1.51)年]、病情分级(部分控制/未控制)(16/5,1/9)、起始C-ACT评分[(20.71±0.85)分,(19.30±1.57)分]、起始FeNO水平[(9.93±7.69)ppb,(32.52±22.70)ppb]方面比较,均差异具有统计学意义(t＝1.72,χ²＝11.98,t＝3.29,t＝4.15;均P<0.05)。显效组患儿起始FeNO水平为(9.93±7.69)ppb,孟鲁司特治疗12周,前后测量结果比较,差异无统计学意义(F＝0.51,P>0.05);无效组患儿治疗前起始FeNO水平为(32.52±22.70)ppb,孟鲁司特治疗12周,前后测量结果之间差异无统计学意义(F＝0.56,P>0.05)。两组患儿孟鲁司特治疗后,PEF%pred、C-ACT评分均提高,6次测量结果之间均差异具有统计学意义(F＝4.63,6.06,50.67,6.09;均P<0.05)。两组患儿孟鲁司特治疗12周期间,除显效组患儿治疗2周时FeNO水平与PEF%pred呈正相关(r＝0.44,P<0.05)外,FeNO水平与PEF%pred、FeNO水平、C-ACT评分无相关性(P>0.05)。 结论FeNO与PEF%pred、C-ACT评分相比暂不具有明显的疗效监测价值,哮喘患儿的病史、病情分级、起始C-ACT评分、起始FeNO水平等因素可能会影响孟鲁司特的疗效。