Investigation Of Prevention And Treatment Of Cardiogenic Stroke And Exploration Of Novel Blood Biomarkers For Cerebral Hemorrhage

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Investigation on prevention and treatment of cardiogenic strokeObjective Physicians' understanding of atrial fibrillation(AF)is a decisive factor in the prevention of cardiogenic stroke.This study aimed to investigate physicians' knowledge,attitude,and behaviors toward cardiogenic stroke,to analyze the influencing factors,to establish a basis for improving neurologists' effective implementation of the guidelines.and to provide data to support departments that develop health policies.Methods This is a cross-sectional study,and a questionnaire survey was used to conduct surveys on the participants' knowledge,attitudes,and behaviors related to cardiogenic stroke.The questionnaire is a self-designed standard structure questionnaire,which has been analyzed in advance of its reliability and validity.Results A total of 611 doctors from 38 hospitals were responded to this survey.The mean of the total score of the questionnaire was 21.59 ± 3.559(total score of the questionnaire was36),and the mean scores of knowledge,attitude and practice were 6.86 ± 1.70,6.13 ± 1.35,8.59 ± 2.21,respectively.The doctor' s knowledge,practice,and total scores were positively correlated with the education level and the workplace.The influencing factors that affect doctors' knowledge,attitudes,and practice scores including education level,professional title,working years,hospital grade,and hospital location.or the best treatment of atrial fibrillation,more than 80% of physicians choose anticoagulant therapy.For patients with atrial fibrillation and cerebral infarction,physicians think Warfarin is the preferred drug as high as93.8%.Among the anticoagulant drugs ever used by clinicians,the use rate of Warfarin is93.8%,but the use rate of direct oral anticoagulants is insufficient.The use of direct oral anticoagulants is related to the educational level and the geographical location of the hospital.Bleeding risk is the first reason influencing clinicians' choice of Warfarin,accounts for 88.9%.97.7% of the clinicians recommend patients with Warfarin regularly monitoring the INR,but the frequency of monitoring is inconsistent.The neurologists' awareness rate of CHADS2 score,CHA2DS2-VASc score,and HAS-BLED score was 79.4%,73.5%,and 62.4%,respectively.However,the utilization rate of three scoring scheme was 39.1%,29.5%,and20.0%,respectively.For the CHADS2 score,hospital location and screening experience were factors negatively associated with both awareness rate and utilization rate,whereas education was factor positively associated with utilization rate.For CHA2DS2-VASc score,hospital location(OR=3.197 vs.3.363),training experience(OR=3.232 vs.2.669),screening experience(OR=1.321 vs.1.746),and professional title(OR=1.565)were factors negatively associated with awareness rate and utilization rate,whereas education(OR=0.667)was factor positively associated with utilization rate.For HAS-BLED score,training experience(OR=3.126 vs.2.985),screening experience(OR=1.558 vs.1.183)and hospital location(OR=1.954)were factors negatively associated with awareness rate and utilization rate,whereas working years(OR=0.676)was factor positively associated with awareness rate.Clinicians have a high willingness to learn about AF,but the proportion of hospitals carry out appropriate training is low.Conclusions There is still a big gap in neurologists' knowledge and practice of atrial fibrillation,especially regarding the guiding principles for the selection of anticoagulant drugs,there are still some gaps.Neurologists have a positive attitude towards anticoagulation therapy and a strong willingness to learn,but lack corresponding training.To improve the management of atrial fibrillation,continuous professional training of doctors must be strengthened.Part ? Exploration of new blood biomarkers in patients with cerebral hemorrhage based on bioinformaticsObjective The pathological damage mechanism of cerebral hemorrhage is complicated,and there is currently a lack of effective therapeutic drugs and biological warning models.Our research is aimed to explore and analyze the expression profile of peripheral blood m RNA of patients with cerebral hemorrhage in public databases to determine the HUB gene associated with cerebral hemorrhage as a therapeutic target,and provide a basis for clinical verification.Methods Download the transcriptome sequencing data of the dataset GSE125512 from the GEO database,screen differentially expressed genes(DEGs)in cerebral hemorrhage,use gene ontology(GO)to annotate DEGs,and use Kyoto Encyclopedia of Genes and Genomes(KEGG)to analyze the signal path of DEGs.Weighted gene co-expression network analysis(WGCNA)was used to screen the key functional modules,and CIBERSORT immune cell infiltration was used to analyze the distribution of immune cell subsets in the peripheral blood of patients with cerebral hemorrhage.In addition,a protein-protein interaction(PPI)network was constructed through DEGs to clarify the relationship between different DEGs,and to select HUB genes with significant interactions.Finally,take the intersection with the HUB gene screened by WGCNA,as a candidate molecule for the next step of verification.Results Among the 218 DEGs,there are 204 differentially expressed m RNAs,including 51up-regulated genes and 14 down-regulated genes.The biological functions of DEGS related to intracerebral hemorrhage mainly involve inflammation and immune response.The cell signaling pathways that were activated 72 hours after the onset of intracerebral hemorrhage identified by KEGG analysis included malaria and PI3K-Akt signaling pathways.CIBERSORT immune cell infiltration analysis showed that compared with 24 hours after the onset of cerebral hemorrhage,the highest content of immune cells was neutrophils at 72 hours after the onset of cerebral hemorrhage,followed by T cells,CD4+ memory resting cells and macrophages,and the onset of cerebral hemorrhage After 72 hours,the content of macrophages was significantly higher than that of 24 hours after the onset,but the content of other immune cells was not significantly different.The PPI network constructed with STRING contains a total of 146 nodes and 446 edges,and finally 13 genes were selected as HUB genes.WGCNA analysis finally identified 15 modules,and 12 HUB genes were identified in the modules with the most significant clinical features.Conclusions Through the analysis of the data set GSE125512,72 hours after intracerebral hemorrhage,there is a significant difference in gene expression compared with 24 hours after intracerebral hemorrhage.The relevant biological pathways are mainly related to inflammation and immune system activation,and the highest content of immunity after intracerebral hemorrhage are neutrophils,and the immune cells with the greatest difference between 72 hours and 24 hours after the onset are macrophages.The enriched meaningful signal pathway is the PI3K-AKT signal pathway.Through PPI and WGCNA analysis,we finally get There are 25 meaningful HUB genes.Part ? Validation of new biomarkers in blood of patients with cerebral hemorrhage based on bioinformaticsObjective Through bioinformatics analysis,the HUB gene related to cerebral hemorrhage was determined,and the expression was verified in clinical patients,and the potential small molecule compounds for the treatment of cerebral hemorrhage were determined.Methods Collected 12 cases of cerebral hemorrhage,cerebral infarction,and control group for clinical verification of HUB gene,and collected peripheral blood samples of patients 72 hours after onset.The final number of verification genes was 21.The RT-q PCR method was used to verify the m RNA levels of 21 HUB genes in the peripheral blood of the three groups of patients.For the genes with obvious differences in expression,the protein levels were further verified by ELISA and WB methods and explored.Correlation with patients' clinical indicators.GSEA enrichment analysis was performed on the successfully verified gene data set GSE125512 again,and meaningful biological processes were obtained.The Drugbank database is used to predict the target drugs for the validated meaningful genes.Results RT-q PCR results showed that among the 21 candidate genes,the m RNA levels of CLEC4 E,DEFA4,OLFM4,ELANE,SUCNR1,and PPBP were higher in the cerebral hemorrhage group than in the control group.The ELISA method confirmed that,at the protein level,the levels of DEFA4 and OLFM4 in the cerebral hemorrhage group were higher than those in the cerebral infarction group and the control group.The WB method was used to verify that the expression of OLFM4 increased in the cerebral hemorrhage group,which was higher than that of the cerebral infarction group and the control group.Further analysis showed that no matter in the cerebral hemorrhage group or the cerebral infarction group,the white blood cell count,neutrophil count,lymphocyte count was not statistically significant in the DEFA4 and OLFM4 expression groups.In addition,whether in the cerebral hemorrhage group or the cerebral infarction group,the expression of DEFA4 and OLFM4 were not significantly different from the NIHSS score at admission and the m RS score at 30 days after onset.In the DEFA4 high expression group,a total of 1 meaningful biological process was enriched,which was a defense response to Gram-negative bacteria.In the OLFM4 high expression group,three meaningful biological processes were enriched,namely:defense response to Gram-negative bacteria,antibacterial humoral immune response,and defense response to fungi;in the OLFM4 low expression group,and also enriched in 3meaningful biological processes,namely smooth muscle cell apoptosis process,positive regulation of muscle cell apoptosis process and response to nerve growth factor.Using the Drugbank database to predict target drugs,there is one drug targeting DEFA4,which is oxaprazine,and there is another drug targeting OLFM4,which is olmesartan.Conclusions Through bioinformatics analysis,we finally verified 6 genes with differential expression(CLEC4E,DEFA4,OLFM4,ELANE,SUCNR1,PPBP)among the 21 candidate genes,and verified that two of them are in the brain at the protein level.The hemorrhage group is highly expressed(DEFA4,OLFM4).It is worth noting that these six different molecules have participated in the collective immune inflammatory response to varying degrees,and their specific roles and mechanisms in cerebral hemorrhage need to be further studied.In addition,we also predicted drugs that use these two molecules as therapeutic targets through the database,and obtained oxaprazine and olmesartan,which may be used as late-stage therapeutic drugs,providing ideas and directions for further research in the later stage.