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Objective To evaluate the inflammatory pattern and the interferon (IFN)-γ in the bronchial secretion of asthma patients in response to acute cold bronchoprovocation. Material and methods We enrolled 42 patients with asthma. We assessed asthma by Asthma Control Test, the lung function by spirometry before and after the bronchodilator test, followed by collecting induced sputum. The next day, we collected exhaled breath condensate (EBC) and conducted a 3-minute isocapnic hyperventilation with cold air (IHCA), followed by collecting spontaneously produced sputum. Results Group 1 included 20 patients with cold airway hyperresponsiveness (CAHR), and group 2 included 22 patients without CAHR. In both groups, a high level of neutrophils in bronchial secretion was observed before and after IHCA. In response to IHCA, the number of epitheliocytes in the sputum decreased to a greater extent in patients of group 1. The baseline epitheliocytes and the concentration of IFN-γ after IHCA had an inverse relationship (r = −0.60; P = 0.017). The baseline IFN-γ in EBC before and after IHCA was lower in group 1. Airway response to cold exposure directly correlated with IFN-γ levels after IHCA (Rs = 0.42; P = 0.014). Conclusion In asthma patients with CAHR, there is a relationship between the persistence of mixed inflammation and the level of IFN-γ in the bronchi. IFN-γ in response to IHCA is decreased with increased cytokine utilization during cold bronchospasm, which is accompanied by the mobilization of neutrophils and the shift in the cytokine spectrum of the respiratory tract towards the T helper cells (Th) 1 immune response.

The scientific literature does not provide enough information on whether bronchial hyperresponsiveness to hypoosmotic stimulus in patients with asthma can lead to more pronounced disturbances of regional lung ventilation.Aim. to characterize lung inflation in asthma patients with osmotic airway hyperresponsiveness.Methods. The lung inflation was studied by body plethysmography, as well as by three-dimensional volumetry, planimetry, and multispiral CT densitometry in 24 patients (group 1) with persistent mild asthma and osmotic airway hyperresponsiveness, identified by the bronchoprovocation test with inhalation of distilled water (IDW) (the average ДРБУ1 was —21.1 ± 3.2%). The comparison group (group 2) consisted of 49 patients with no response to IDW (the average ДББУ1 was —3.7 ± 0.5%; p = 0.00001).Results. Group 1 had lower lung function (FEVj was 83.6 ± 4.5%; FEF50 was 58.1 ± 5.8%) at baseline in comparison with the group 2 (96.7 ± 2.2%, p = 0.0042 and 75.5 ± 2.2%, p = 0.016, respectively) and higher indices of lung inflation at body plethysmography (RV was 153.2 ± 12.5 and 127.5 ± 4.0%, respectively; p = 0,027; RV/TLC was 128.8 ± 5.5 and 109.9 ± 2.8%, respectively; p = 0.015). According to three-dimensional volumetry, the indicators of expiratory lung inflation (526.0 ± 117.8 vox) and average residual inflation of both lungs (13.1 ± 2.6 vox) in group 1 were significantly higher than in group 2 (301.5 ± 55.8 vox, р < 0.05 and 9.1 ± 1.6 vox,р < 0,05, respectively). The patients with osmotic airway hyperresponsiveness also showed higher values of the expiratory area in the middle zone (235.3 ± 29.4 and 149.2 ± 14.9 pix, respectively; p = 0.00 47) and the lower zone (292.3 ± 37.9 and 178.6 ± 18.6 pix, respectively; p = 0.0034) of the lungs.Conclusion. Asthma patients with osmotic airway hyperresponsiveness have lung hyperinflation with impaired lung ventilation predominantly in the middle and lower zones.

Introduction. Currently, COVID-19, which is rapidly spreading around the world in the form of a pandemic, is a serious public health problem that poses a significant epidemiological and medico-social threat to the population and its quality of life, and affects all spheres of public life and economy. Aim. To assess the degree of influence of the pandemic of the new respiratory viral infection COVID-19 on the level of registered primary morbidity by classes of diseases, including the class "Diseases of the respiratory system" in the territory of the Far Eastern Federal District.Materials and methods. To implement the tasks of the study, a complex of analytical, epidemiological and statistical studies was carried out. Methods used: epidemiological analysis and monitoring, statistical, comparative and correlation analysis and methods for analyzing time series. Epidemiological and statistical assessment of the incidence of the population was carried out on the basis of ICD-10 according to the data of the state statistical monitoring of the incidence of the population of the Russian Federation for 2019-2020 using the database of the Ministry of Health of the Russian Federation, the Central Research Institute of Organization and Informatization of Healthcare of the Ministry of Health of the Russian Federation.Results. The pandemic of the new respiratory viral infection COVID-19 significantly changed the dynamics of the registered morbidity in the adult population, which dropped sharply in all ICD-10 classes, most intensively in the classes "Diseases of the endocrine system" (by 27.8%), "Diseases of the blood and hematopoietic organs" (by 22.6%), "Neoplasms" (by 21.1%), "Diseases of the circulatory system" (by 18.5%), with the exception of the class "Diseases of the respiratory system", the level of primary morbidity for which in the territory of the Far Eastern Federal District in 2020 increased by 23.9%. The most common and severe clinical manifestation of COVID-19 is bilateral pneumonia, the incidence of which among the adult population in the Far Eastern Federal District increased 2.7 times, and in the territory of the Russian Federation – 3.8 times. At the same time, the incidence of pneumonia in the child population decreased by 36.4%. Along with the growth of acute forms of respiratory pathology during the COVID-19 pandemic in the Far Eastern Federal District, the level of registered primary morbidity of chronic respiratory diseases decreased: chronic bronchitis (by 22.3%), bronchialasthma (by 20.8%), COPD (by 3, 6%).Conclusion. The negative dynamics of the growth of the registered morbidity of the population can have negative consequences due to the late detection and untimely diagnosis of chronic forms of pathology, including respiratory diseases, their progression and the development of life-threatening complications of the disease and, as a consequence, an increase in mortality from these causes of the population, who did not receive timely and adequate medical assistance.

Purpose Mucus secretion is excessively increased in airway epithelial cells in pathological states. This process is related to the cytoskeleton and the increase in exocytosis sites, but the movement of secreted molecules and how secretion increases remain unclear. In this study, we examined the potential role of myristoylated alanine rich C kinase substrate (MARCKS) and the cortical actin-binding protein cortactin in airway mucin secretion. Also we investigated the effect of activated Cdc42-associated kinase 1 (ACK1) in this process. Methods Human airway epithelial cells were treated with neutrophil elastase (NE) after treatment with siRNA to specifically knock down MARCKS, ACK1 and cortactin expression. The expression and localization of cortactin and MARCKS were observed by western blotting and immunofluorescence, and the phosphorylated forms of MARCKS, cortactin and ACK1 were detected. The interaction of cortactin and ACK1 was analyzed by coimmunoprecipitation. MUC5AC protein expression was measured by ELISAs. Results Phosphorylated cortactin was highly expressed, mainly at the cell membrane, after NE stimulation, and phosphorylated MARCKS was mainly expressed in the cytoplasm. Coimmunoprecipitation revealed that ACK1 and cortactin interacted with each other. Knockdown of MARCKS suppressed phosphorylation of cortactin, while cortactin siRNA had no significant effect on MARCKS activation. Knockdown of MARCKS, cortactin and ACK1 by siRNA attenuated the phosphorylation of cortactin and reduced MUC5AC secretion. Conclusion These results suggest that both cortactin and MARCKS are involved in MUC5AC secretion by increasing F-actin polymerization and translocation and that MARCKs and ACK1 play an important role in the activation of cortactin.

Introduction. Hypoalimentation with the formation of protein-energy malnutrition can reduce the body’s resistance to acute respiratory infections (ARI). Soy and pumpkin can be a promising source of normalization of the diet.Aim. To study the effectiveness of the use of soybean-pumpkin products in the diet of healthy people to increase the resistance of the human body to ARI.Materials and methods. An open, randomized, parallel group study was conducted. Group 1 included 18 people who took a soy-pumpkin drink and dessert for 10 weeks in addition to their usual diet. Group 2 included 17 people on a regular diet. The concentration of vitamin E, cholesterol, α- and β-lipoproteins was determined in blood serum. Diene conjugates and unoxidized lipids were determined in the expired air condensate (EAC) by means of spectrophotometry. Lung function was assessed using spirometry.Results. According to the questionnaire survey, 50% of the participants in group 1 registered a positive dynamics from the regular use of soybean-pumpkin products. Only one participant underwent ARI. In the 2nd group, 7 people (41%) applied for medical help (χ2=4.43; p˂0.05), of which 6 (35%) had ARIs. At the end of the observation, an increase in vitamin E (from 34.1±0.68 to 36.4±0.55 μg/mL, p=0.0036), a decrease in total cholesterol (from 4.85±0.23 to 4.32±0.14 mmol/L, p=0.0013) and β-lipoproteins (from 2.92±0.24 to 2.31±0.18 mmol/L, p=0.0003). At the beginning of the study, in individuals of group 1, there was a relationship between age and the concentration of total cholesterol (r=0.50; p=0.036) and β-lipoproteins (r=0.47; p=0.048), as well as the weight and level of α-lipoproteins (r=-0.49; p=0.041), the concentration of unoxidized lipids (r=0.55; p=0.02), diene conjugates (r=0.48; p=0.043) in EAC. After regular use of soybean-pumpkin products, this correlation dependence was lost.Conclusion. Regular use of soy-pumpkin products for 10 weeks has a beneficial effect on the lipid profile of the blood, increases antioxidant defence, and reduces the susceptibility of a healthy person to ARI.

Since the beginning of the 50s of the last century, numerous classifications of sarcoidosis have been proposed, which to one degree or another satisfied specialists at certain stages of the development of medical technologies. Currently, sarcoidosis of the respiratory system is divided into five stages (from 0 to IV) based on the results of chest Xray examination. Active introduction into clinical practice of computed tomography and its more advanced version – lowdose multislice computed tomography (MSCT) – allows to carry out an effective diagnosis of sarcoidosis, including in outpatient practice, as well as when performing active screening and dynamic monitoring of the course of the disease. On the basis of the studies which were carried out, the dissonance between the progress in technology (from X-ray pattern to MSCT) and the stagnation of the conceptual issues of the classification of sarcoidosis is discussed, which urgently requires its reforming. Firstly, the category of patients with concomitant lesions of the intrathoracic lymph nodes (ITLN) and parenchyma (stage II) has sharply increased, which reflects the increased technological level of radiological diagnostics. As a result, the percentage of stages with isolated lymph node involvement (stage I) and isolated dissemination of the parenchyma (stage III) decreased. The classification based on MSCT data allows to more accurately determine the severity of the disease. This is of therapeutic and prognostic importance, since most patients of stage I generally do not need treatment, while symptomatic stage II requires hormone therapy. One should also take into account the possibility of spontaneous remission, which is observed more often in stage I. The main conclusion is that at present day the principle of staging in the classification of sarcoidosis can no longer satisfy the realities of modern clinics. It is more correct to single out the clinical and radiological forms of sarcoidosis: sarcoidosis of the ITLN, sarcoidosis of the ITLN and lungs, sarcoidosis of the lungs, as well as sarcoidosis of the respiratory system, combined with a single lesion of other organs.

Full functioning of the airway physical barrier depends on cellular integrity, which is coordinated by a series of tight junction (TJ) proteins. Due to airway spasm, edema, and mucus obstruction, positive end-expiratory alveolar pressure (also termed auto-PEEP) is a common pathophysiological phenomenon, especially in acute asthma attack. However, the influence of auto-PEEP on small airway epithelial TJs is currently unclear. We performed studies to investigate the effect of extra pressure on small airway epithelial TJs and its mechanism. The results first confirmed that a novel mechanosensitive receptor, piezo-1, was highly expressed in the airway epithelium of asthmatic mice. Extra pressure induced the degradation of occludin, ZO-1 and claudin-18 in primary human small airway epithelial cells (HSAECs), resulting in a decrease in transepithelial electrical resistance (TER) and an increase in cell layer permeability. Through in vitro investigations, we observed that exogenous pressure stimulation could elevate the intracellular calcium concentration ([Ca2+]i) in HSAECs. Downregulation of piezo-1 with siRNA and pretreatment with BAPTA-AM or ALLN reduced the degradation of TJs and attenuated the impairment of TJ function induced by exogenous pressure. These findings indicate the critical role of piezo-1/[Ca2+]i/calpain signaling in the regulation of small airway TJs under extra pressure stimulation.

Certain transient receptor potential (TRP) channels including TRPM8 and TRPA1 are widely expressed in the respiratory tract and have been shown to be the receptors of cigarette smoke and particulate matter—the main causative factors of chronic obstructive pulmonary disease (COPD). The aim of the study was to investigate the effect of TRPM8 and TRPA1 polymorphisms on COPD predisposition and lung function in COPD patients. The study enrolled 143 COPD patients and 104 smokers with post-bronchodilator forced expiratory volume in one second (FEV1)/forced vital capacity (FVC) > 70%. Lung function was measured by spirometry. TRPM8 and TRPA1 polymorphisms were genotyped by LATE-PCR. None of the polymorphisms significantly influenced COPD predisposition after correction for covariates and multiple testing. Among COPD patients, the TT genotype of TRPA1 rs7819749 was significantly associated with higher degree of bronchial obstruction. In addition, we established that carriers of the C allele of TRPM8 rs11562975 more commonly had post-bronchodilator FEV1 < 60% (OR 3.2, 95%CI (1.14–8.94), p = 0.03) and revealed the effect of TRPA1 rs959976 and TRPM8 rs17865682 on bronchodilator response in COPD. Thus, the obtained results suggest possible involvement of TRPM8 and TRPA1 in COPD pathogenesis, indicating the necessity to further investigate their functional role in this pathology.

Background: Airway hyperresponsiveness is related to the intensification of the lipid peroxidation (LP) in asthma. We aimed to study the effect of smoking on the LP status of asthma patients with EIB. Methods: 62 subjects (36.2±1.3 years) with EIB (drop of FEV1 -21.9±2.0%) were divided into 2 groups: the 1st included 30 non-smokers, the 2nd - 32 smokers (8.2±1.5 pack/year). The contents of conjugated dienes and non-oxidized lipids in the exhaled breath condensate were analyzed before and after 6-minute submaximal exercise on treadmill, using ultraviolet spectra of lipid extracts absorption and evaluated in terms of optical density at 233 nm (E233) and 206 nm (E206), respectively. Results: In group 2, FEV1 was lower (84.6±2.7 vs. 92.0±2.5%; p 0.05) in the2nd group led to a change in E233/E206 after exercise: 0.39±0.09 (p>0.05) vs. 0.38±0.05 (p Conclusion: Smoking in asthmatics with EIB leads to changes in the LP, which is accompanied by a deterioration of airway patency and a decrease of the response to dosed exercise.

Background: Features of bronchial inflammation that contribute to the development of cold airway hyperresponsiveness in patients with asthma have been poorly studied. We aimed to investigate the correlation between the neutrophils and airway response to cold stimulus. Methods: In 116 patients with uncontrolled asthma (mean age 43.2±0.8 years), the lung function (FEV1,%), airway response to 3-minute isocapnic hyperventilation with cold air (IHCA), the cell composition of induced sputum (IS, %), the activity of myeloperoxidase (MPO, in pixels), and the degree of structural integrity of cells using the destruction index (DI, relative units) were studied. Asthma control was assessed by ACT questionnaire. Results: The patients were divided into 2 groups based on the number of neutrophils in IS: group 1 (n=52) less than 40%, and group 2 (n=64) more than 40%. The number of eosinophils in IS was 21.2±1.4 vs. 14.3±1.4%, respectively (p=0.001). The baseline FEV1 was 87.3±1.7 and 84.3±2.6% (p>0.05); the FEV1 drop in response to IHCA was -10.9±1.3 and -10.9±1.6%. The level of ACT in group 1 was higher (13.7±0.33 vs. 12.4±0.44 points, p=0.026), the MPO content of granulocytes was lower (81.7±3.6 vs. 135.9±12.1 pixels, p=0.00001), as well as neutrophil DI (0.47±0.02 vs. 0.56±0.04, p=0.047) and DI of the bronchial epithelium (0.52±0.02 vs. 0.63±0.06, p=0.05). The correlation was found between the content of neutrophils in IS and the airway response to IHCA (r=-0.18, p=0.047). Conclusion: An increase in the number of neutrophils with an escalation of their functional activity in patients with uncontrolled asthma is related to the severity of the airway response to the IHCA.

Asthma is a widespread chronic disease of the bronchopulmonary system with a heterogeneous course due to the complex etiopathogenesis. Natural-climatic and anthropogenic factors play an important role in the development and progression of this pathology. The reception of physical and chemical environmental stimuli and the regulation of body temperature are mediated by thermosensory channels, members of a subfamily of transient receptor potential (TRP) ion channels. It has been found that genes encoding vanilloid, ankyrin, and melastatin TRP channels are involved in the development of some asthma phenotypes and in the formation of exacerbations of this pathology. The review summarizes modern views on the role of high and low temperatures in airway inflammation in asthma. The participation of thermosensory TRP channels (vanilloid, ankyrin, and melastatin TRP channels) in the reaction to high and low temperatures and air humidity as well as in the formation of bronchial hyperreactivity and respiratory symptoms accompanying asthma is described. The genetic aspects of the functioning of thermosensory TRP channels are discussed. It is shown that new methods of treatment of asthma exacerbations caused by the influence of temperature and humidity should be based on the regulation of channel activity.

The aim of our study was to evaluate the role of airway cellular inflammation and the lipid peroxidation level in the development of airway hyperresponsiveness (AHR) to inhalation of hypertonic saline (IHS). Methods and Results: The study included the estimation of inflammatory-cellular composition, intracellular concentration of myeloperoxidase (MPO) in induced sputum (IS), serum levels of lipid hydroperoxides (LHP), ceruloplasmin, and vitamin E in 29 patients with asthma and 12 healthy persons. AHR to IHS was assessed by spirometry after 3-min IHS via ultrasonic nebulizer. Patients with asthma had higher indices of leukocytes destruction and cytolysis intensity with the increased leukocyte count in IS. Maximum values of neutrophils cytolysis intensity and leukocytic MPO were found in IS of the patients with AHR to IHS. After the bronchial provocation, serum concentration of LHP was higher in these patients in comparison with the patients without the AHR and control groups. In addition, patients with asthma had lower level of antioxidants than healthy subjects. Conclusion: Marked inflammation involving MPO-activated leukocytes and intensive lipid peroxidation underlie the excessive airway response to IHS.

Background: Role of interferon-gamma (IFNγ) in asthma remains controversial. While certain studies found association of this cytokine with severe asthma and bronchial hyperresponsiveness, the other ones established its rather beneficial effect on mucin secretion and asthma development. Objectives: The aim of the study was to investigate IFNγ levels in exhaled breath condensate (EBC) of asthma patients. Methods: The study included 46 patients with asthma (mostly newly diagnosed - 52% and moderate - 30%) and 23 patients with chronic bronchitis. Mean age of patients was 38.1±1.60 years. Among asthma patients, 78% had uncontrolled disease according to ACQ5. Lung function was tested by spirometry. IFNγ was measured in EBC by BD cytometric bead array method with enhanced sensitivity. Results: Asthma patients had higher level of IFNγ than patients of control group (2654 (1328.9; 3123.9) fg/ml vs. 1872 (1707.8; 2506.3) fg/ml) but the difference was not significant. IFNγ was increased in moderate asthma (2199 (1764.8; 2519) fg/ml) compared to newly diagnosed asthma (1666 (928.8; 2048.5), p=0.01). Moreover, IFNγ was positively correlated with ACQ5 score (R=0.39, p=0.01). In uncontrolled asthma concentration of the cytokine was more than two times higher compared to patients with partially or fully controlled disease (2807 (1512.9; 3123.9) fg/ml vs. 1158 (995.5; 1589.6) fg/ml, p=0.02). 10-days treatment with oral glucocorticoids decreased IFNγ from the initial (2654 (1328.9; 3123.9) fg/ml) to “normal” values (1790 (1124.4; 3070.1) fg/ml, p=0.08). Conclusions: Our observations suggest detrimental effect of IFNγ in asthma pathogenesis. The study was supported by RFBR (#18-54-53019).

Background: Airway hyperresponsiveness (AHR) to osmotic stimuli is common and clinically relevant in asthma. AQP5 is a membrane water channel that may be functionally implicated in airway inflammation and hypersecretion. Objectives: The aim was to assess the effect of AQP5 SNPs on osmotic AHR in asthma patients. Methods: The study enrolled 274 patients with mild-to-moderate asthma, mean age 36.2±0.67 years. The patients underwent bronchoprovocation with 3-min inhalation of distilled water (hypoosmotic stimulus) and hypertonic saline aerosol (hyperosmotic stimulus). Lung function was measured by spirometry before and after each challenge. AQP5 SNPs (rs3759129, rs3736309, rs296756, rs1964676) were genotyped by LATE-PCR. Results: Association with hypoosmotic AHR was found for rs3759129 and rs296756. C allele of rs3759129 was protective against hypoosmotic AHR as AC heterozygotes had less profound response to the challenge compared to AA carriers (ΔFEV1 -1.4 (-6.3; 2.33)% vs. -5.4 (-12.0; -1.0)%, p=0.001; ΔFEF25-75 -1.0 (-18.0; 14.0)% vs. -9.1 (-21.1; 1.2)%, p=0.009). G allele of rs296756 in homozygous state also had protective effect. AA and AG (but not GG) carriers showed marked reduction in airway patency (ΔFEV1 -5,5 (-12,0; -0,8)% vs. -0,2 (-4,5; 3,4)%, p=0.004; ΔFEF25-75 -9,6 (-22,0; 2,3)% vs. 2,0 (-8,9; 9,0)%, p=0.008, for AA and GG genotypes, respectively). rs3759129 also influenced hyperosmotic AHR: all patients with the AHR had AA genotype while 27% of patients without the AHR had AC genotype (p=0.006). Conclusions:AC genotype of rs3759129 and GG genotype of rs296756 exert a protective effect on hypoosmotic AHR. AC genotype of rs3759129 also protects asthma patients from hyperosmotic AHR.

The aim of the research was to study the state of the bronchial mucosa epi-thelium in relation to the severity of clinical manifestations in severe uncon-trolled asthma depending on the pattern of inflammation and the presence of cold airway hyperresponsiveness. Materials and methods. In 48 patients with severe uncontrolled asthma, there were assessed asthma symptoms, clinical signs of cold airway hyperre-sponsiveness, and lung function; the samples of slides were analyzed in the cytological examination of the sputum; the degree of damage to epithelial cells and granulocytes was estimated using the total cell destruction index (CDI). Results. According to the analysis of sputum cytograms, the patients were divided into two groups: group I (22 patients) included persons with eosin-ophilic inflammation pattern (31.0±3.1% of eosinophils and 22.0±2.2% of neutrophils), group II (26 patients) was with mixed inflammation pattern (7.2±1.4 and 71.8±4.2%, respectively). The patients of group II had lower disease control according to Asthma Control Test (ACT; 12.1±0.7 and 17.8±0.2 points, respectively; р

To explore the role of cortical actin-binding protein (cortactin) in shear stress-induced mucin (MUC) 5AC secretion in human airway epithelial cells and the effect of phosphorylation of cortactin at different sites. Methods: HBE16 airway epithelial cells were cultured, and then transfected with mutation carrier, such as pEGFP-N1-cortactin (Cort), pEGFP-N1-Cort-Y421A, pEGFP-N1-Cort-Y470A and pEGFP-N1-Cort-Y486A. The cells were divided into a normal control group, a shear stress group, a shear stress + pEGFP-N1 group, a shear stress + PEGFP-N1-Cort group, a shear stress + pEGFP-N1-Cort-Y421A group, a shear stress + pEGFP-N1-Cort-Y470A group, and a shear stress + pEGFP-N1-Cort-Y486A group. The shear stress were set at 4 dynes/cm2. The levels of MUC5AC protein and mRNA in cells and culture supernatant were assayed with enzyme-linked immunosorbent assay (ELISA) and real-time PCR. The cortactin and phosphorylated cortactin were detected by Western blot. F-actin was stained by fluorescein isothiocyanate (FITC)-phalloidin. Results: There was an obvious increase of phosphorylated cortactin in cells exposed to 4 dynes/cm2 of shear stress for 30 min, which reached climax at 2 hours concomitant with elevation of MUC5AC protein production and mRNA expression in the different experiment groups (all P0.05). Compared with single shear stress-stimulated group, MUC5AC in supernatant was increased obviously, and the distribution of F-actin in cytomembrane was also increased in the pEGFP-N1-Cort group (both P0.05), while there were no changes in the MUC5AC protein and mRNA levels in cytoplasm. Compared with the shear stress+pEGFP-N1-Cort group, the MUC5AC protein in the culture supernatant was decreased, and the polymerization of F-actin at cell membranes were also attenuated in the shear stress+pEGFP-N1-Cort-Y421A group and the shear stress + pEGFP-N1-Cort-Y470A group (both P0.05), while there was no significant effect in the shear stress + pEGFP-N1-Cort-Y486A group (P0.05). Conclusion: Cortactin is involved in shear stress-mediated MUC5AC secretion in human airway epithelial cells, and the phosphorylated site of Tyr421 and Tyr470 may play an important role in it.目的：了解皮层肌动结合蛋白(cortical actin-binding protein，又称cortactin)在人气道上皮细胞黏蛋白(mucin，MUC)5AC分泌中的作用及不同磷酸化位点对其的影响。方法：培养人气道上皮细胞HBE16，以皮层肌动结合蛋白不同磷酸化位点突变载体转染细胞，并分为空白对照组、剪应力刺激组、剪应力+增强绿色荧光蛋白质粒(enhanced green fluorescent protein plasmid， pEGFP)-N1组、剪应力+pEGFP-N1-cortactin (Cort)组、剪应力+pEGFP-N1-Cort-Y421A组、剪应力+pEGFP-N1-Cort-Y470A组及剪应力+pEGFP-N1-Cort-Y486A组，剪应力设定为4 dynes/cm2。采用酶联免疫吸附法(enzyme-linked immunosorbent assay，ELISA)、real-time PCR法测定转染前后各组细胞及培养上清中MUC5AC蛋白和mRNA水平，Western印迹检测各组皮层肌动结合蛋白及磷酸化皮层肌动结合蛋白含量；异硫氰酸荧光素(fluorescein isothiocyanate，FITC)标记的鬼笔环肽染色观察丝状肌动蛋白(filamentous actin，F-actin)的分布情况。结果：4 dynes/cm2剪应力刺激30 min后，细胞中磷酸化皮层肌动结合蛋白表达增加，在2 h时表达水平最高；剪应力刺激2 h后，MUC5AC mRNA表达增强，细胞及培养上清中MUC5AC的含量与空白对照组相比，各组均显著增加(均P0.05)；与剪应力刺激组相比，剪应力+pEGFP-N1-cortactin(Cort)组的细胞培养上清液中MUC5AC蛋白含量显著增加，F-actin在胞膜的分布也明显增多(均P0.05)，而胞质内MUC5AC蛋白及MUC5AC mRNA水平变化不大。与剪应力+pEGFP-N1-Cort组相比，剪应力+pEGFP-N1-Cort-Y421A组、剪应力+pEGFP-N1-Cort-Y470A组培养上清液中的MUC5AC蛋白含量有明显降低，F-actin在胞膜的聚集也有所减少(均P0.05)，而剪应力+pEGFP-N1-Cort-Y486A组中MUC5AC含量变化不明显(P0.05)。结论：皮层肌动结合蛋白参与了剪应力诱导产生的人气道上皮细胞MUC5AC蛋白分泌，Tyr421和Tyr470位点磷酸化在其中可能起重要的作用。.

Acute or chronic cold exposure exacerbates chronic inflammatory airway diseases, such as chronic obstructive pulmonary disease (COPD) and asthma. Cold-inducible RNA-binding protein (CIRP) is a cold-shock protein and is induced by various environmental stressors, such as hypothermia and hypoxia. In this study, we showed that CIRP gene and protein levels were significantly increased in patients with COPD and in rats with chronic airway inflammation compared with healthy subjects. Similarly, inflammatory cytokine production and MUC5AC secretion were up-regulated in rats following cigarette smoke inhalation. Cold temperature-induced CIRP overexpression and translocation were shown to be dependent on arginine methylation in vitro. CIRP overexpression promoted stress granule (SG) assembly. In the cytoplasm, the stability of pro-inflammatory cytokine mRNAs was increased through specific interactions between CIRP and mediator mRNA 3'-UTRs; these interactions increased the mRNA translation, resulting in MUC5AC overproduction in response to cold stress. Conversely, CIRP silencing and a methyltransferase inhibitor (adenosine dialdehyde) promoted cytokine mRNA degradation and inhibited the inflammatory response and mucus hypersecretion. These findings indicate that cold temperature can induce an airway inflammatory response and excess mucus production via a CIRP-mediated increase in mRNA stability and protein translation.

The effect of single nucleotide polymorphisms (SNP) of TRPV4 gene on the development of airway hyperresponsiveness (39.7% of cases) in response to the decrease in osmolarity under inspiration of distilled water aerosol was studies in 189 patients with uncontrolled bronchial asthma. rs6606743 SNP was found to significantly contribute to the development of osmotic airway hyperresponsiveness. Analysis of the dominant genetic model revealed substantial prevalence of AG + GG genotype frequency in the group of patients with asthma with osmotic hyperresponsiveness in comparison with the patients who had negative response to bronchoprovocation. In addition, carriers of GG or AG genotypes had significantly more profound decrease of lung function parameters in relation to A homozygous patients.

Mucin 5AC (MUC5AC) is a highly O-glycosylated mucin secreted by human bronchial epithelial cells during pulmonary inflammatory diseases. T antigen, a component of the MUC5AC glycans, is the product of the O-glycosylation transferase T-synthase and its chaperone Cosmc. Since the expression of Cosmc is mediated by signaling pathways and inflammatory factors affecting mucin O-glycosylation, we analyzed the impact of neutrophil elastase (NE)-mediated Cosmc and T antigen expression in BEAS-2B cells derived from human bronchial epithelial cells. The expression of Cosmc and T antigen in human lung tissue was analyzed by immunohistochemistry. Cellular immunohistochemistry and western blot analysis demonstrated elevated expression of T antigen in BEAS-2B cells after NE stimulation. Altered Cosmc expression in BEAS-2B cells after NE stimulation was analyzed by confocal microscopy, western blot analysis and quantitative RT-PCR. To assess the biological implications of Cosmc function for T-synthase activity and T antigen synthesis after NE stimulation, BEAS-2B cells were transfected with shRNA to silence the expression of Cosmc. The changes in signaling pathways were analyzed by western blotting. The expression of Cosmc and T antigen increased in lung tissue exposed to chronic inflammation. The expression of Cosmc and T antigen increased in NE-stimulated BEAS-2B cells. NE induced increases in T antigen expression and T-synthase transferase activity in BEAS-2B cells expressing Cosmc, highlighting the importance of Cosmc in the relationship between NE and T antigen. Cosmc and phosphatidylinositol-3-kinase (PI3K) played important roles in the signaling pathway that stimulated hyperexpression of T antigen.

Chronic inflammatory lung diseases accompanied by Gram-negative bacteria infection are characterized by excessive mucin production. Lipopolysaccharide (LPS), the major endotoxin released from Gram-negative bacteria, is a potent inflammatory agonist for mucin overproduction. In this study, we sought to examine whether the toll-like receptor (TLR)-responsive microRNA miR-155 plays a role in LPS-provoked induction of mucin 5AC (MUC5AC) and the potential role of suppressor of cytokine signaling 1 (SOCS1) involved in this process. We found that LPS increased the expression of MUC5AC in association with TLR4-dependent miR-155 induction. The suppression of miR-155 by antagomir led to an excessive production of SOCS1, thereby downregulation of MUC5AC production. Collectively, these data imply that miR-155 is involved in LPS-induced MUC5AC overproduction through a TLR4-dependent manner and thereby the downregulation of SOCS1.

Background: The definition of the personality traits involved in the formation of patients’ adherence is practically important. Aim: to determine the influence of personal characteristics of patients with asthma on their medication adherence. Methods: 93 patients with mild (n=60) and moderate (n=33) asthma at the age of 18-58 years were examined with the help of Minnesota Multiphasic Personality Inventory (MMPI). 85% of them were women. Results: 54 patients were adherent to the medication (group 1), and 39 patients didn’t fully perform medical prescription or change the therapeutic regimen by themselves. Analyzing the data of psychological tests, significantly higher values of the average ratings of MMPI scores De (depression), Pt (psychasthenia), Sc (schizophrenia) and Si (social introversion) (p 70 T-points). Soreness and individuality of patients of the 2nd group makes it difficult to predict their speech and behavior. Conclusion: The presence of anxiety and depressive disorders, introversion and soreness tendency and personal identity of asthma patients have a negative influence on their medication adherence. To achieve the asthma control it should be taken into account when predicting the clinical course and then it should be corrected on time.