Objective To observe the eotaxin expression of rat airway smooth muscle cells ( ASMCs) induced by serum from asthmatic rats, and explore the possible mechanism. Methods ASMCs isolated fromrat tracheas were cultured in vivo. Then they were treated with serum from asthmatic rats, or treated with serum and dexamethasone simultaneously. The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction. The expression of cAMP in ASMCs was examined by radioimmunoassay. Results After the treatment with sensitized serum, the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [ ( 107. 09 ±7. 12) ng/L vs. ( 0. 63 ±0. 56) ng/L, P lt; 0. 05; 1. 39 ±0. 04 vs. 0. 05 ±0. 01, P lt;0. 05] , and the level of cAMP in ASMCs was significantly lower compared with the control group [ ( 17. 58 ±3. 62) ng/L vs. ( 32. 39 ±3. 36) ng/L, P lt; 0. 05] . After intervened by the sensitized serum and dexamethasone simultaneously, the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serumalone [ ( 64. 18 ±4. 04) ng/L and 0. 77 ±0. 19] . The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs ( r = - 0. 788, P lt; 0. 01) . Conclusions There is anautocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum. Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
ObjectiveTo explore the application of pulmonary ventilation and perfusion imaging (V/P SPECT/CT) in quantitative evaluation of ventilation and perfusion function and its potential value in guiding local treatment of lung in patients with asthma.MethodsA total of 20 patients with asthma were included in this study. All patients underwent V/P SPECT/CT and pulmonary function test, and symptoms were assessed by the ACT questionnaire. Patients were graded for degree of airway obstruction according to V/ P SPECT/CT image visual scoring criteria. The comprehensive lung function (%) of the patients was quantitatively evaluated by combining the ventilation and perfusion defect of each lung segment in V/P imaging. The correlation between the degree of airway obstruction, comprehensive lung function, pulmonary function test and ACT score was analyzed.ResultsV/P SPECT/CT imaging can be used to grade the degree of airway obstruction in asthma patients (0-3 grade). Airway obstruction grading by V/P SPECT/CT visual score was associated with predictive forced expiratory volume in one second (FEV1%pred) of patients (r=–0.74, P<0.001). V/P SPECT/CT can also comprehensively evaluate ventilation and perfusion function in patients with asthma, and comprehensive lung function measured by this method was also correlated with FEV1%pred (r=0.629, P=0.003). V/P SPECT/CT can be used to quantitatively analyze the percentage of ventilation and perfusion function in each lung lobe. Compared with V/P SPECT/CT results, the CT volume overestimates the contribution in the upper lobes, and underestimates the lower lobes contribution to overall function.ConclusionsV/P SPECT/CT can be used as a new method to directly reflect the degree of airway obstruction in patients with asthma. Moreover, it can comprehensively and quantitatively evaluate the ventilation and perfusion function of asthma patients. V/P SPECT/CT can also be used to evaluate lobe function in patients with asthma, helping to identify the heterogeneity of changes in pulmonary function in patients with asthma, and has potential value for future treatment targeting specific areas of the lung.
Objective To evaluate the efficacy of Budesonide / formoterol to control asthma under real-life conditions. Methods A multi-center, open label, non-interventional study was conducted. Asthma control after 12 week therapy with Budesonide/ formoterol was assessed by Asthma Control Questionnaire( ACQ) and modified Asthma Control Questionnaire ( ACQ5) . Results A total of 360 asthma patients were recruited, including 228 adult patients and 132 child patients. After 12 weeks’ therapy, all the patients’medium value of ACQ was decreased significantly from 2. 03 ( adults 2. 20, children 1. 74) at baseline to 0. 60 ( adults 0. 78, children 0. 29) ( P lt; 0. 0001 ) , and the medium value of ACQ5 was also decreased significantly from2. 4 ( adults 2. 24, children 1. 76) at baseline to 0. 47 ( adults 0. 62, children 0. 20) ( P lt;0. 0001) . Conclusion Budesonide / formoterol is effective in asthma treatment, by which most asthma patients obtain and maintain clincal control.
Objective To investigate the relevance and changes of mucosal immunity in asthma rats’lung, nose and intestine. Methods Twenty Wistar rats were randomly divided into a normal group and an asthma group. Asthma rat model was established by sensitization and challenge with ovalbumin. CD4 + ,CD8 + , eotaxin protein and its mRNA in rats’lung tissues, rhinal and intestinal mucosa were measured by immunohistochemical methods and situ hybridization. The content of sIgA in bronchoalveolar lavage fluid ( BALF) , nasopharyngeal washings and intestinal mucus supernatant were detected by enzyme-linked immunosorbent assay. Results Compared with the normal group, the levels of CD4 + , CD8 + in rats’lung tissues, rhinal and intestinal mucosa, the expression of eotaxin protein and mRNA in rats’lung tissues, the content of sIgA in nasopharyngeal washing, and the expression of eotaxin protein in intestinal mucosa were significantly higher in the asthma group( P lt; 0. 05) . There were no significant differences of other indices between the two groups. In the normal group, the eotaxin protein expression had a negative correlationbetween lung tissue and rhinal mucosa( r = - 0. 572, P = 0. 008) , and a positive correlation between intestinal and rhinal mucosa( r=0. 638, P =0. 002) . The eotaxin mRNA expression had a positive correlation between lung tissue and rhinal mucosa( r= 0. 502, P = 0. 024) , and a positive correlation between intestinaland rhinal mucosa( r=0. 594, P =0. 006) . In the asthma group, such a correlation was not found except the eotaxin protein expression which had a negative correlation between lung tissue and intestinal mucosa( r =- 0. 448, P = 0. 048) . Conclusions Mucosal immunity in lung, nose and intestine remains a dynamic balance. The balance of mucosal immunity is destroyed in asthma.
Objective To investigate the expression of RNP（alpha defensins） in guinea pigs with bronchial asthma and the influences of P2Y2 receptors on the expression. Methods Seventy-two adult male guinea pigs were randomly divided into a control group （group A） and an asthma group （group B）. The asthma model was established by sensitizing with ovalbumin injection intraperitoneally and challenging with ovalbumin inhalation. Then the A and B groups were divided into following subgroups,ie,the saline groups （A1/B1），ATP groups （A2/B2），and Suramin groups （A3/B3），in which the animals were intervented with saline，ATP，and suramin respectively. Total pulmonary resistance （RL） was measured.Total and differential cell counts in bronchoalveolar lavage fluid （BALF） were measured. Pathological changes of lung were observed under light microscope using HE staining. The plasma levels of RNP and IL-8 were measured by ELISA. The protein and mRNA expressions of RNP and P2Y2 were detected by immunohistochemistry and RT-PCR. Results The RL increased significantly as much as 10% in group B compared with group A. Pathological study revealed that luminal narrowing of bronchus and inflammatory cells infiltration in the asthma group. The total cell and polymorphonuclear neutrophil （PMN） counts in BALF in group B1 were significantly higher than group A1 and B3，but lower than group B2 （Plt;0.05）,but no significant difference among group A1，A2，or A3 was found. The RNP expression in plasma and lung tissue in group B1 was higher than group A1 and B3，but lower than group B2 （Plt;0.05）. The RNP expression in plasma and lung tissue was lower in group A1 than group A2，but higher than group A3（Plt;0.05）. The IL-8 expression in plasma in group B1 was significantly higher than group A and B3，but lower than group B2 （Plt;0.05） but no significant difference among group A1，A2，or A3 was found（Pgt;0.05）. RNP protein expressed predominantly in lung tissues，especially where the PMN infiltrated. The expression of RNP mRNA were consistent with the RNP protein expression in all groups. P2Y2 mRNA expression in group A1 was lower than group A2，and higher than group A3 （Plt;0.05）.P2Y2 mRNA expression in group B1 was lower than group B2 but higher than group B3 （Plt;0.05）. The linear correlation analysis showed that RNP had no significant correlation with PMN and IL-8 in group A，and was positively correlated with P2Y2 mRNA （Plt;0.05） while RNP was positively correlated with PMN，IL-8，and P2Y2 mRNA in group B. Conclusion RNP expression increases in bronchial asthma guinea pigs and may be related to P2Y2 receptors.
ObjectiveTo explore the expressions of nerve growth factor (NGF) and leukemia inhibitory factor (LIF) in both asthmatic mice and respiratory syncytial virus(RSV)-infected mice,explore if there is a same neurogenic mechanism between ashtma and RSV infection,in order to find a new treatment target for asthma. MethodsOne hundred healthy Balb/c inbred mice were randomly divided into a control group,a RSV group,an asthma group,an asthma with RSV group,and a dexamethasone group. The lung tissue pathology was observed by hematoxylin-eosin staining(HE). The quantitative analysis of NGF mRNA and LIF mRNA of lung tissue was detected by RT-PCR. The expression of NGF protein and LIF protein was detected by immunohistochemical method. ResultsUnder light mocroscope,there were alveolar septum widening,alveolar epithelium swelling,and interstitial edema in the RSV group. There were widen alveolar septum,narrowed bronchial lumen,thicken bronchial wall and a large number of inflammatory cells infiltration around the small blood vessels,alveolar and bronchioles both in the asthma group and the asthma with RSV group,with the latter being more serious. Compared with the RSV group,the inflammation was relieved significantly in the dexamethason group. There were mRNA and protein expressions of NGF and LIF in all groups, which were highest in the asthma with RSV group,then the RSV group and the asthma group,and lowest in the dexamethasone group. ConclusionsThe expressions of LIF and NGF in the lung of mice after RSV infection and futher increase when combined with asthma. Dexamethason can inhibit the expression of NGF and LIF to some extent.
The number of clinical guidelines developed and published in different countries is increasing worldwide. Too many guidelines do not remain in regular use, even though the aim is to implement them in clinical practice. The scientific validity and reliability of the guidelines need to be reviewed. Here is a case presented to show how to optimally use the evidence-based guideline to improve clinical decision making.
ObjectiveTo explore the composition of intestinal microbiota between patients with fixed airflow obstruction asthma, reversible airflow obstruction asthma, and healthy control, and analyze the correlation between key differential bacterial distribution and clinical characteristics. MethodsFifteen patients with fixed airflow obstruction asthma (FAO) and 13 patients with reversible airflow obstruction asthma (RAO) were included, along with 11 matched healthy control subjects. Clinical data were collected, and lung function tests and induced sputum examination were performed. Blood and stool samples were tested to compare the gut microbiota status among the groups, and analyze the relationship between gut microbiota abundance and patients' blood routine, IgE levels, lung function, and induced sputum. Results The dominant bacterial compositions were similar in the three groups, but there were differences in the abundance of some species. Compared to the RAO group, the FAO group showed a significant increase in the genera of Bacteroides and Escherichia coli, while Pseudomonas was significantly decreased. The phylum Firmicutes was negatively correlated with the course of asthma, while the phylum Bacteroidetes and genus Bacteroides were positively correlated with the asthma course. Bacteroidetes was negatively correlated with Pre-BD FEV1/FVC, Pseudomonas was positively correlated with Pre-BD FEV1, Escherichia coli was negatively correlated with Post-BD FEV1/FVC, and Bacteroides was negatively correlated with Post-BD MMEF. The class Actinobacteria and the order Actinomycetales were negatively correlated with peripheral blood EOS%, while the order Enterobacteriales and the family Enterobacteriaceae were positively correlated with peripheral blood IgE levels. Furthermore, Actinobacteria and Actinomycetales were negatively correlated with induced sputum EOS%. Conclusions There are differences in the gut microbiota among patients with fixed airflow obstruction asthma, reversible airflow obstruction asthma, and healthy individuals. Bacteroides and Escherichia coli are enriched in the fixed airflow obstruction asthma group, while the Firmicutes are increased in the reversible airflow obstruction asthma group. These three microbiota may act together on Th2 cell-mediated inflammatory responses, influencing the process of airway remodeling, and thereby interfering with the occurrence of fixed airflow obstruction in asthma.
Objective To evaluate the effects of different inspiratory flow waveforms on the respiratory function of patients with severe exacerbation of asthma during mechanical ventilation. Methods Twenty-one patients with severe exacerbation of asthma were ventilated with square waveform and decelerating waveform respectively for 30 minutes when the tidal volume was set at 6 mL/ kg, 8 mL/ kg and 10 mL/ kg in ICUof Zhejiang Hospital of Integrated Traditional Chinese and Western Medicine fromJanuary 2006 to December 2007. Meanwhile shunt fraction ( Q·S /Q·T ) , dead space value ( VD/VT ) , airway peak pressure ( Ppeak ) , plateau pressure ( Pplat) ,intrinsic positive end-expiratory pressure( PEEPi) and arterial blood gas analysis were measured. Results The Q ·S /Q·T in the decelerating waveformgroup was less than that of the square waveform group( P lt;0. 05) when tidal volume was set at 6 mL/ kg. When tidal volume was set at 10 mL/ kg, PEEPi and VD /VT in the decelerating waveform group were higher than those of the square waveform group. On the contrary, the Ppeak was lower than that of square waveform group( P lt; 0. 05) . Conclusion For patients with severe exacerbation of asthma treated with mechanical ventilation, decelerating waveform is preferable at low tidal volume( 6 mL/ kg) , and square waveform is preferable at high tidal volume( 10 mL/kg) .
Objective To investigate the feasibility of dendritic cells ( DCs ) as vector of immunotherapy through intratracheal injection. Methods The DCs obtained from the bone marrow of BALB/ c mice were cultured and isolated with CD11c-positive magnetic beads. Then DCs were overloaded with ovalbumin peptide 323-339 ( OVA 323-339) for 24 hours. The mice in the DC-OVA group were intratrachelly injected DCs overloaded with OVA 323-339 in dose of 2 ×106 cells per mouse. The mice in thenegative control group were intratracheally injected with DCs untreated by OVA 323-339. On the second day,all mice were challenged with 1% OVA in PBS lasting for five days. The asthma animal model established by classic method was used for the positive control. Pathologic changes in lung and cell numbers in bronchoalveolar lavage fluid ( BALF) were assayed 24 hours after challenged. Results Just like the lung tissues from the mice asthma models, the lung tissues from the mice instilled with DCs overloaded with allergen OVA 323-339 showed extensive inflammatory cells infiltration, most of which were eosinophils, neutrophils and lymphocytes. The lung tissues in the DC group showed no obvious inflammation. There were more cells in BALF in the DC-OVA group than that in the DC group. OVA-specific IgE in serum from the DC-OVA group was not significantly different from that in the mice asthma models [ ( 48. 22 ±4. 76) U/mL vs. ( 52. 75 ±4. 03) U/mL, P gt;0. 05] . Conclusion DCs overloaded antigen has the ability of transferring of antigen effectively and may be used as vectors of immunotherapy.