Screening Of Serum Early Warning Protein Markers In Uremic Patients With Cerebral Hemorrhage

Objective: To compare and identify the differentially expressed proteins in the serum of patients with uremia and uremia combined with cerebral hemorrhage,and to screen "specific protein molecules" as biomarkers for diagnosis and early warning of patients of uremia combined with cerebral hemorrhage.Methods: Collecting 6 cases of uremia patients' serum and 6 cases of uremia patients with cerebral hemorrhage in the emergency department of the First Affiliated Hospital of Nanchang University,and collect 6 healthy adult serum specimens,and divide the specimens into 3 groups:(1)uremia patients group;Patients with cerebral hemorrhage;(3)control group of healthy adults.The serum of each group of patients was subjected to protein extraction and enzymatic hydrolysis,and then the appropriate amount of peptides were taken for chromatographic separation using the Nano n-flow Easy nLC 1200 chromatography system(Thermo Scientific).)Perform DDA(Data Dependent Acquisition)mass spectrometry,and the resulting LC-MS /MS raw data is imported into the search engine Sequest HT in Proteome Discoverer software(version 2.2)for database retrieval.Using bioinformatics analysis and its role in the pathophysiological mechanism of uremia intracerebral hemorrhage,screen and identify differentially expressed proteins as biomarker candidate proteins,and verify the screened differential proteins by PRM method.Results: Through significant difference analysis,a total of three groups of experimental results were obtained,of which 77 were significantly differentially expressed proteins,71 were up-regulated proteins,and 6 were down-regulated proteins in the uremia and normal control groups;the uremic intracerebral hemorrhage was significantly differentially expressed in the normal control group There were 43 proteins,38 up-regulated proteins,and 5 down-regulated proteins.There were 26 differential proteins in the comparison group of patients with uremic cerebral hemorrhage and uremia,all of which were down-regulated proteins.By categorizing and comparing the three sets of data,a total of 67 differential proteins were obtained.According to cell biology analysis,6 differentially expressed proteinswere selected as candidate marker proteins,including thioredoxin,coagulation factor XIIIB chain,[Cu-Zn] SOD and PHPT1 of these four candidate proteins showed the same trend in the results of isotopic labeling quantitative proteomics and PRM quantitative results,which has statistical significance.Conclusions : Isotope-labeled proteomics combined with PRM validation method revealed that TXN?F13B?[Cu-Zn] SOD and PHPT1 play an important role in the pathophysiological mechanism of uremia complicated with cerebral hemorrhage and can be used as a biomarker for the early warning of serious complications of uremia cerebral hemorrhage.