Clinical Observation On Treatment Of CAA With TCM Kidney-tonifying Method And Primary Screening Of Exosomes Differential Proteins In CAA

ObjectiveTo observe the TCM symptoms and peripheral blood changes of chronic aplastic anemia(CAA)patients of kidney-yang deficiency and kidney-yin deficiency types before and after treatment with TCM kidney-tonifying method,so as to clarify the clinical efficacy of the method,explore the differences in efficacy between the above two types,and screen potential specific biomarkers of CAA with the help of exosomes,explore the possible correlation between them and the mechanism of the disease.MethodsA total of 68 patients with CAA(34 cases of kidney-yang deficiency and34 cases of kidney-yin deficiency)according to the criteria of diagnosis,exclusion and TCM syndrome classification,and 10 healthy volunteers were selected as healthy control group.Before treatment,20 initial diagnosed CAA patients and 5 healthy control group volunteers were selected between groups,their 3ml of peripheral blood was collected for the extraction of exosomal protein.The differential proteins with potential marker value were screened and analyzed by liquid chromatography-mass spectrometry tandem analysis(LC-MS/MS)and quantitative technology of targeted protein group based on mass spectrometry(PRM).Sixty-eight patients with CAA were treated with TCM kidney-tonifying method combined with conventional basic treatment for 6 months.The general data,TCM syndrome scores and blood changes before and after treatment were observed and compared between the two types of patients,and the efficacy was determined and compared.Results1.The treatment of patients with CAA by TCM kidney-tonifying method had considerable clinical efficacy,with a total clinical efficiency of 75.81%and a total efficiency of 90.32% in improving the TCM syndrome score.2.Self comparison before and after treatment with TCM kidney-tonifying method in both syndrome types: The total scores of TCM syndromes of kidney-yang deficiency and kidney-yin deficiency syndrome types were decreased(P<0.05),The hemogram of patients of kidney-yang deficiency syndrome type were increased(P<0.05).The hemoglobin and platelet count of kidney-yin deficiency syndrome type patients were increased(P<0.05),there was no significant difference in reticulocyte count,leukocyte count and neutrophil count before and after treatment(P>0.05).3.Comparison between groups before and after treatment: There was no significant difference in clinical effective rate or TCM total scores improvement between kidney-yang deficiency syndrome and kidney-yin deficiency syndrome(P>0.05).The improvement in hemoglobin,reticulocyte count,neutrophil count and platelet count of kidney-yang deficiency syndrome was better than that of kidney-yin deficiency syndrome(P<0.05).There was no significant difference in the improvement of white blood cell count between the two syndrome types(P>0.05).4.The exosomes detection show that CAA has 82 up-regulated differential proteins and 62 down-regulated differential proteins;Further under the verification analysis of PRM,the three most significant differential proteins were finally screened out,they were CETP、 TFRC and COL1A1.Conclusions1.TCM kidney-tonifying method can effectively improve the TCM symptoms of CAA patients and increase the baseline of peripheral hemogram,with exact clinical effect.2.Patients of kidney-yang deficiency type of CAA are better than those of kidney-yin deficiency type in improving the TCM syndrome and peripheral hemogram,and have better clinical response to TCM kidney-tonifying method.3.The exosomes proteins extracted from the peripheral blood of CAA patients are combined with LC-MS/MS and PRM analysis to screen and obtain differential proteins with potential marker values represented by CETP,TFRC and COL1A1.