| Bronchial asthmas is a widespread chronic disease involved in many inflammtory cells and factors, this makes patients to have airway hyperreponsiveness (AHR) to varies excitative factors . With deepen investigation to airway inflammation, a close attention to the neurogenic inflammation in airway which mediated by sensory neuropeptides secreted by sensory nerve. Substance P is an important kinds of sensory neuropeptides, SP has the effects to asthma , such as, smooth muscle contraction, microvascular leakage, increased mucus secretion, inflammatory cell activation and causing airway AHR,so we think there is a close relation between SP and asthmatic symptoms. Induce sputum technique has been developed in several years, it has safety, utility, noninvasion, and wonderfull repetition . Inflammatory cells and factors in induced sputum are better than that of BALF, so we detect the concentration SP in asthmastic induced sputum .Objects and Methods1 .Clinical data; there were 34 asthmastic children in acute episode period, 30 asthmastic children in remission period, 18 health children in the control subjects.2.Methods:(1)The collection of induced sputum: induced sputum was collected respectively, in acute episode period group, in remission period group and in control group, the method of sputum induction is refer to Pin's. The inductionis was carried out with an aerosol 3% ,4% ,5% of hypertoil saline by an ultrasonic nebulizer, for three periods of 7 min, total <30 min. After inhalating ,the child is asked to cough and try to expectorate sputum into a cleaning plat. Selecting qualification sputum 1.5ml, 1ml sputum is added in a cold test tube including 10% EDTA 50ul,trasylol 800u,and was stored at -70℃ for future analysis. The specimen is treated with sputum protectional liquor ( produced the china medicinal college) to break up the mucus and disperse the cells, and centrifuged at 1900r/min for 10 min, cytospons were airdried and stained with HE.(2)Treatment of sputum: The sputum samples were diluted with acetic acid(4% ,PH4.0)to a final volume of 20ml, homogenized, and centrifuged at 40,000 x g for 30 min. The obtained supernatant was loaded on reversed -phase CIS cartridges. After washing with 10ml of 4% acetic acid and 10ml distilled water, SP was eluted with 2ml of 80% acetonitrile in 0. 1% trifluoroacetic acid. Elutes were concentrated by spin vacuum evaporation, lyophilized and stored at -70@ until assay.(3)Measurement of SP concentration: the elutes were dissolved with 0. 3ml assay buffer (50nM phosphate buffer, PH7. 2,containning 3. 7mg/ml EDTA and 0.5% bovine serumal bumn). A total of 0.2ml of the dissolved preparation was subjected to further radioimmunoassay for SP. RIA for SP was performed by the instruction of SP.3.Statistical Analysis;Data are expressed as mean ?SE. Comparisons of the SP concentration in hypertonic saline - induced sputum among groups were made with students paired t test (twwo -tailed). A P value of <0. 01 was considered significant.Result1. The different counts of cells in induce sputum in the acute episode group , Remission period group and normal control group.The percentage of neutrophil in induce sputum in the acute episode group compared with both Remission period group and control group has significantly increased,p <0. 01. The percentage of eosino-philic in the acute episode group , Remission period group compared with the control group has significantly increased,p <0.01.2.The SP concentration in induce sputum in the acute episode group , Remission period group and control group.The SP concentration in induce sputum in the acute episode group compared with both Remission period group and control group has significantly increased,p <0.01. The SP concentration in induce sputum in the Remission period group compared with the control group hasnot significantly increased, p > 0.05.Discussion(一) The investigation of SP in children asthma 1.sensory neuropeptide and asthma;The pathogenesis of bronchial asthma is very complicate...
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