Study For The Determination Method Of The Total SAA Content And SAA Glycosylationand The Relationship Between Activity And Content

Objective: Spleen disorder salivary amylase (sAA) acid stimulation/previous activityratios lower than normal, as the dialectical TCM spleen deficiency diagnostic referenceindex is used for traditional Chinese medicine basic and clinical research, however, spleendisorder salivary amylase activity changes mechanism remains elusive, the paper mainlyaffect sAA activity sAA content and glycosylation level studies, the establishment WBdetermination sAA glycosylation methods and the total activity of sAA and sAAglycosylation and the correlation between the total analyzed in order to TCM spleendeficiency sAA activity before and after acid stimulation mechanism of protein expressionchanges in the direction of relevant research methodologies reference.Method:1Specification of citric acid to stimulate saliva collection method;2collected before and after stimulation citric healthy adults, children and spleen child salivasamples, saliva pH, salivary flow rate, primarily by EPS-G7sAA activity rate method, BCAprotein detection kit saliva total protein concentration, Western Blot determination sAAtotal and glycosylation sAA were calculated specific activity of sAA and sAA activity andglycosylation total content correlation analysis.Results:(i) using0.4mol/L1×1cm citric acid filter stimulated salivary flow rate(stimulation:1.4944±0.5692mL/min; stimulation:0.8528±0.4047mL/min) and sAAactivity ratio (stimulation:1.5571±0.4585; stimulation:1.7246±0.4527) increased mostsignificantly (P <0.05);(ii) when measured sAA activity, EPS-G7rate method, coefficientof variation (CV) under control in less than5%, while the iodine-starch method The CV is more than10%, and half in the30-48%;(iii) Western Blot technique DAB chromogenicmethod glycosylation sAA content of the total sAA20%-30%;(iv) in healthy adults andchildren, acid stimulation, saliva pH, salivary flow rate, sAA activity, sAA concentration,glycosylation sAA concentrations have increased specific activity, including children, toincrease more obvious (P <0.05). These indicators acid stimulation after stimulation withacid ratio of small children than adults. In addition, adult acid stimulation before and afterhigh sAA activity than children. Acid stimulation, adult saliva total protein concentrationdecreased significantly (P <0.05), while the children are on the increase. Children acidstimulated spleen v, saliva pH, the rate of flow of saliva, sAA activity, sAA concentration,glycosylation sAA concentrations have increased specific activity, but these indicators acidstimulation ratio of the acid stimulation, in addition to total protein concentration in salivaand sAA concentrations did not change significantly, the spleen children are less thanhealthy children. In addition, acid-stimulated spleen children stabbed before sAA activity,saliva total protein concentration, sAA and glycosylation sAA concentration and specificactivity were greater than in healthy children acids corresponding values before stimulation.(Vi) in healthy adults, spleen children sAA activity sAA and glycosylation sAAconcentration correlation and healthy children sAA activity and glycosylation sAAcorrelations are acid concentration after stimulation than acid stimulation, healthy adults,sAA activity and sAA correlation between the concentration of the acid stimulation thanafter acid stimulation, in which healthy children: pH ago sAA activity and sAAconcentrations higher correlation (R=0.697), acid stimulation sAA activity andglycosylation sAA concentrations than the correlation high (R=0.503).Conclusion:⑴successfully regulate the citric acid filter stimulate saliva collectionmethods that select0.4mol/L1×1cm citric acid filter stimulate1min,2min whole salivacollected after stimulation; establish more stable measuring sAA activity EPS-G7ratemethod; successfully established WesternBlot determination sAA content and contentglycosylation approach for sAA activity change mechanism provides a methodologicalreference.⑵Children acid deficiency before stimulation sAA activity than healthychildren, acid stimulation after/before sAA activity ratios lower than healthy children, and clinical spleen deficiency dialectical theory consistent with further evidence of theexperimental method is accurate and reliable.⑶acid content before and after stimulationsAA activity correlation with differences, and showed a significant positive correlation.sAA glycosylation level of content and are the factors affecting their activity.