The monitoring of airway inflammation has assessed in bronchial asthma directly by sputum examination, and indirectly by measurements in peripheral blood. To investigate the diagnostic value of these two methods, we compared nitric oxide (NO) metabolites, eosinophils, and eosinophil cationic protein (ECP) in sputum and blood in patients with asthma and control subjects. Sputum and serum were obtained from fifteen patients with asthma, and then were examined before anti-asthma treatment, including steroid preparations. ECP was measured by fluoroimmunoassay. NO metabolites were assayed by using modified Griess reaction. Asthmatic patients, compared with control subjects, had significantly higher level of NO metabolites, higher proportion of eosinophils, and higher levels of ECP in sputum. Asthmatic patients, compared with control subjects, however, had significantly higher number of eosinophils, and were at higher levels of ECP in blood. FEV1, FEV1 /FVC was negatively correlated with sputum eosinophils. The area under receiver operating characteristic(ROC) curve showed that eosinophils in sputum are significantly accurate markers than NO metabolites in sputum and blood. These findings suggest that the proportion of eosinophils in sputum have more accurate diagnostic marker of asthmatic airway inflammation than NO metabolites in sputum in differentiating asthmatic patients from control subjects.
Adult ; Area Under Curve ; Asthma/*blood/*metabolism ; Blood Proteins/*metabolism ; Comparative Study ; Eosinophils/*metabolism ; Female ; Fluoroimmunoassay ; Human ; Inflammation ; Male ; Nitrates/metabolism ; Nitric Oxide/blood/*metabolism ; Nitrites/metabolism ; ROC Curve ; Ribonucleases/blood/*metabolism ; Sputum/*metabolism

AIMTo explore the expression of heme oxygenase-1 (HO-1) in the peripheral blood mononuclear cell (PBMC) and relationship to ventilatory function in asthmatic patients.

METHODSEighteen asthmatic patients and eighteen healthy subjects were selected. HO-1 protein levels in PBMC were measured by immunohistochemical staining and PBMC HO-1 mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR), blood carbon monoxide Hb (COHb) percent value, serum total IgE concentration and pulmonary ventilatory function were observed in asthmatic patients and healthy subjects.

RESULTSThe percentage of cells in immunohistochemical staining positive staining of HO-1 were significantly higher in asthmatic patients (41.7 +/- 7.44%) compared with that of healthy subjects (10.5 +/- 4.36%, P < 0.01), the optical densities of PBMC HO-1 mRNA were higher in asthmatic patients (26.05 +/- 4.14) compared with that of healthy subjects (10.82 +/- 4.26, P < 0.01). The relation analysis showed PBMC HO-1 protein levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.56, -0.51, P < 0.01, respectively) and positive relation with COHb percent value, serum total I gE concentration (r = 0.80, 0.48, P < 0.05, respectively), and PBMC HO-1 mRNA levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.65, -0.67, P < 0.05, respectively) and positive relation with COHb percent value, serum total IgE concentration (r = 0.85, 0.62, P < 0.01, respectively).

CONCLUSIONThe expression of PBMC HO-1 protein and mRNA are increased significantly in asthmatic patients, HO-1 may play a significant role in the pathogenesis of asthma. The expression of HO-1 has relation with severity of asthma.

Adult ; Asthma ; blood ; pathology ; Case-Control Studies ; Female ; Heme Oxygenase-1 ; blood ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged

OBJECTIVEEosinophils play a pivotal role in asthmatic airway inflammation. We previously found a significantly high expression of Slingshot-1L (SSH-1L) in peripheral eosinophils in acute exacerbations of asthma. Objective To investigate the expression and localization patterns of SSH-1L in peripheral blood eosinophils of asthmatic patients and their changes after treatment with inhaled corticosteroids.

METHODSWe recruited 4 outpatients with acute exacerbations of asthma who received no previous corticosteroid treatment and 1 healthy volunteer. From all the subjects 30 ml peripheral venous blood samples were collected before and after a 3-month treatment with inhaled fluticasone. The eosinophils were isolated, purified and counted, and the expressions of SSH-1L in the eosinophils were examined by RT-PCR and Western blotting. The localization of SSH-1L phosphatases in the peripheral eosinophils was detected by immunofluorescence assay in one patient.

RESULTSSSH-1L phosphatases distributed diffusely in the cytoplasm, especially dense near the membrane of the peripheral eosinophils. Glucocorticoids treatment resulted in a significant reduction in both the SSH-1L mRNA expression (0.7403∓0.1124 vs 0.4101∓0.0363, P=0.001) and SSH-1L protein expression (0.3410∓0.1337 vs 0.1543∓0.0551, P=0.039).

CONCLUSIONA high expression of SSH-1L in peripheral eosinophils in acute exacerbations of asthma may play a role in the activation and migration of eosinophils. The efficacy of inhaled corticosteroids in asthma control might be partly attributed to a down-regulated expression of SSH-1L.

Adult ; Aged ; Asthma ; blood ; drug therapy ; Eosinophils ; metabolism ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Middle Aged ; Phosphoprotein Phosphatases ; metabolism

Adolescent ; Asthma ; blood ; CD40 Ligand ; blood ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Immunoglobulin E ; blood ; Interleukin-4 ; blood ; T-Lymphocytes ; metabolism

OBJECTIVETo explore the role of T lymphocytes activation co-stimulation pathway and T lymphocyte subset activation in asthma pathogenesis.

METHODSThe blood samples were taken from 35 asthma children (including 22 male and 13 female, age 11 months-9 years) and 31 normal children (including 19 male and 12 female, age 8 months-12 years). Direct immunofluorescence flow cytometry was used to detect the CD86 mean fluorescence intensity (MFI) on CD(14)(+) cell, CD(19)(+) cell percentage and CD19 and CD86 double positive cell percentage in PBMC activated by LPS. ELISA was used to detect the levels of IL-4, IL-13, IFN-gamma in culture supernatants of PBMC stimulated with PHA and plasma total IgE level.

RESULTS(1) The CD86 MFI on CD(14)(+) cell in asthma group was elevated significantly (31.8 +/- 9.2 vs 23.5 +/- 6.4, P < 0.01). (2) CD(19)(+) cell percentage and CD19 and CD86 double positive cell percentage in PBMC stimulated with LPS in asthma group were increased significantly (13.5 +/- 4.0 and 4.6 +/- 2.0 vs. 8.2 +/- 3.0 and 2.3 +/- 1.4, respectively, P < 0.01). The plasma total IgE level [186.6 (64.3 - 723.6)] was higher than that of control's [41.95 (0.9 - 115.2)]. (3) The levels of IL-4 and IL-13 in supernatants stimulated with PHA increased significantly [34.2 (2.8 - 70.0) and 1 239.0 (378.3 - 2 929.0) vs. 9.7 (2.0 - 46.2) and 683.2 (128.8 - 1 560.8) respectively, P < 0.01, P < 0.05]; and the level of IFN-gamma decreased significantly [12.16 (1.6 - 44.8) vs. 26.7 (2.1 - 99.5) P < 0.05]. (4) In asthma group there was a significantly positive correlation of CD86 MFI on CD(14)(+) cell with CD19 and CD86 double positive cell percentage (r = 0.644), there was a significantly positive correlation of CD(19)(+)CD(86)(+)cells percentage with plasma total IgE (r = 0.537); there was a significantly positive correlation of CD(19)(+)cells percentage with the levels of IL-4 and IL-13 (r = 0.607, 0.540 respectively); a significantly positive correlation of CD(19)(+)CD(86)(+) percentage cells with the levels of IL-4 and IL-13 was found (r = 0.617, 0.678, respectively).

CONCLUSIONS(1) The expression of CD86 on APC cells increased in children with acute asthma. (2) The response to mitogens of B lymphocyte and T(H2) but not T(H1) lymphocyte were increased significantly. The results suggest that CD86 and imbalance of T(H) subset may play an important role in asthma pathogenesis.

Antigens, CD ; blood ; Asthma ; blood ; metabolism ; B-Lymphocytes ; metabolism ; B7-2 Antigen ; Child ; Child, Preschool ; Cytokines ; blood ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Direct ; Humans ; Immunoglobulin E ; blood ; Infant ; Membrane Glycoproteins ; blood ; Monocytes ; metabolism

OBJECTIVETo study the serum level of interleukin-17A (IL-17A) and its expressions in the lung, spleen and thymus in asthmatic mice.

METHODSIn 14 normal BALB/c female mice and 14 asthmatic mice, the changes in the airway pathology and the cell proportion in the bronchoalveolar lavage fluid (BALF) were observed. The serum level of IL-17A and IL-17A expressions in the tissue homogenates of the lung, spleen and thymus of the mice were detected by sandwich enzyme-linked immunosorbent assay (ELISA).

RESULTSThe airway inflammation in the asthmatic mice was characterized mainly by eosinophil and neutrophil infiltration, which was not observed in the normal control group. Serum IL-17A levels and IL-17A expressions in the lung, spleen and thymus of the asthmatic mice were significantly higher than those in the normal control group (P<0.01). In the asthmatic mice, IL-17A expression in the lung tissues was positively correlated with the percentages of neutrophils (r=0.693, P=0.040) and eosinophils (r=0.733, P=0.030) in the BALF.

CONCLUSIONIL-17A is highly expressed in the serum, lung, spleen and thymus of asthmatic mice. IL-17A may be one of the major cytokines involved in exacerbation of bronchial asthma, and is probably associated with the recruitment of neutrophils and eosinophils into the airways.

Animals ; Asthma ; blood ; etiology ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Interleukin-17 ; blood ; metabolism ; Mice ; Mice, Inbred BALB C ; Random Allocation

Recently the prevalence of both asthma and obesity have increased substantially in many countries. The aim of this study was to evaluate the role of retinol-binding protein (RBP) 4 in childhood asthma and its association with atopy markers, pulmonary function, and bronchial hyperresponsiveness in relation to obesity. We studied 160 children between the ages 6 to 10 yr, including 122 asthmatics and 38 controls. The body mass index, pulmonary function tests, and methacholine challenge tests were measured on the same day. Total eosinophil count, serum total IgE, serum eosinophil cationic protein, and serum RBP4 were measured in all subjects. There was no difference in serum RBP4 levels between the asthmatics and the control group. In all subjects or subgroups, serum RBP4 was not associated with total eosinophil count, serum total IgE, serum eosinophil cationic protein, or PC20. There was no relationship between serum RBP4 and pulmonary function in female asthmatics. Forced expiratory volume in 1 second/forced vital capacity (FVC) and forced expiratory flow between 25% and 75% of FVC contributed to serum RBP4 in male asthmatics. Our findings show an association between RBP4 and pulmonary function in prepubertal male asthmatics. This relationship may indirectly affect the high prevalence of childhood asthma in males.
*Asthma/blood/immunology ; Bronchial Hyperreactivity/blood/immunology ; Bronchial Provocation Tests ; Child ; Female ; Humans ; Male ; Obesity/blood/immunology ; Retinol-Binding Proteins, Plasma/*metabolism

OBJECTIVETo investigate the changes of the levels of galectin-3 (Gal-3) in serum and bronchoalveolar lavage fluid (BALF) of children with asthma whose have different serum levels of 25-hydroxyl-vitamin D₃[25(OH)D₃].

METHODSFifty children with asthma between January 2013 and December 2014 were enrolled as the asthma group, and they were classified into 25(OH)D₃sufficient (n=7), insufficient (n=12) and deficient subgroups (n=31) according to the serum levels of 25(OH)D₃. Twenty children with abnormal airway or tracheal foreign bodies served as the control group. The levels of 25(OH)D₃, Gal-3 and total IgE in serum and Gal-3 levels in BALF were measured using ELISA.

RESULTThe serum levels of 25(OH)D₃in the asthma group were lower than in the control group (P<0.05). The 25(OH)D₃deficient subgroup displayed the highest percentages of neutrophils, eosinophils and epithelial cells in BALF, followed by the 25(OH)D₃insufficient subgroup and the 25(OH)D₃sufficient subgroup (P<0.05). The percentages of neutrophils, eosinophils and epithelial cells in BALF in the three subgroups were all higher than in the control group (P<0.05). In children with asthma, serum levels of 25(OH)D₃were negatively correlated with the percentages of neutrophils, eosinophils and epithelial cells in BALF (r=-0.683, -0.795 and -0.670 respectively; P<0.05); and a negative correlation was also seen between serum 25(OH)D₃levels and serum Gal-3 and total IgE levels (r=-0.759 and -0.875 respectively; P<0.05).

CONCLUSIONSThe children with asthma have low serum levels of 25(OH)D₃. 25(OH)D₃and Gal-3 may be involved in the airway inflammation and the development of asthma.

Asthma ; etiology ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Child ; Child, Preschool ; Female ; Galectin 3 ; analysis ; blood ; physiology ; Humans ; Immunoglobulin E ; blood ; Infant ; Male ; Vitamin D ; analogs & derivatives ; blood ; physiology

BACKGROUNDAirway symptoms in asthma are related to decrease of epinephrine secretion, which may be ascribed to elevated nerve growth factor (NGF) in the organism. The aim of this study was to monitor the neuroendocrine alteration in the adrenal medulla of asthmatic rats.

METHODSSixteen rats were randomly divided into two groups (n = 8), control group and asthma group, and the asthmatic rats were sensitized and challenged with ovalbumin (OVA). The levels of NGF, epinephrine and norepinephrine in serum were detected by enzyme linked immunosorbent assay (ELISA), the NGF expression in adrenal medulla was detected by immunohistochemistry, and the changes in the ultrastructure of the adrenal medulla was observed by electron microscopy.

RESULTSThe NGF expression was increased in asthmatic rats compared with control rats. Compared with control rats, the results indicated that the epinephrine level was decreased in asthmatic rats, but no significant difference was found in norepinephrine levels. We found more ganglion cells in the adrenal medulla of asthmatic rats than in control rats, with NGF immunostaining mainly located in these ganglion cells. Electron microscopic images showed the density of chromaffin granula decreased and there was shrunken nucleolemma in the adrenal medullary cells of asthmatic rats.

CONCLUSIONThe innervation of the adrenal medulla is changed in asthmatic rats, and it may contribute to the epinephrine decrease in asthma.

Adrenal Medulla ; metabolism ; ultrastructure ; Animals ; Asthma ; blood ; metabolism ; Bronchoalveolar Lavage ; Enzyme-Linked Immunosorbent Assay ; Epinephrine ; blood ; Male ; Microscopy, Electron, Transmission ; Nerve Growth Factor ; blood ; metabolism ; Norepinephrine ; blood ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVEThis study examined the levels of exhaled nitric oxide (eNO) and peripheral blood eosinophils (EOS) as well as the correlation between the two markers in children with bronchial asthma (AS),AS complicated by allergic rhinitis (AS/AR) and chronic cough variant asthma (CVA), in order to explore the value of eNOS detection in children with AS.

METHODSThe eNO level was measured using light-emitting electrochemical photometry in 12 children with AS, 29 children with AS/AR and 10 children with CVA. Peripheral blood EOS was counted by blood cell counter (Coulter JT). Forced expiratory volume in one second (FEV1) was assessed by lung function measurement. Thirty children without atopic disease and acute respiratory infection as well as without a family history of atopic diseasea served as the control group.

RESULTSThe levels of eNO and blood EOS in the AS, the AS/AR and the CVA groups were significantly higher than those in the control group (p<0.01). The AS/AR group showed increased levels of eNO (50.3 + or - 6.7 ppb) and EOS (5.9 + or -4.2 x 109 ) compared with the AS (30.5 + or - 8.8 ppb and 4.2 + or - 3.2 x 109 respectively) and the CVA groups (26.0 + or - 3.2 ppb and 3.7 + or - 6.9 x 109 respectively) (p<0.05). There were no significant differences in eNO and EOS levels between the AS and the CVA groups. The eNO level was positively correlated with the EOS level (r=0.51, p<0.05), but not with FEV1 (r=0.144, p>0.05) in the AS group.

CONCLUSIONSNO is highly expressed in children with symptoms of atopy and can reflect the levels of eosinophilic airway inflammation in children with AS.

Adolescent ; Asthma ; blood ; physiopathology ; Breath Tests ; Child ; Child, Preschool ; Eosinophils ; physiology ; Female ; Forced Expiratory Volume ; Humans ; Male ; Nitric Oxide ; metabolism