The Role And Potential Mechanism Of Neutrophil Counts In Predicting The Hospital Prognosis Of Patients With Phlegm-blood Stasis Type Of Standford Type B Acute Aortic Dissection

Objective This study was aimed to assess the value of neutrophil counts in predicting the risk of death for hospitalized patients with phlegm-blood stasis type of Standford Type B acute aortic dissection(AAD).The microarray was used to detect the differentially expressed miRNAs in patients with phlegm-blood stasis type of Standford Type B AAD and healthy controls in peripheral neutrophils,using bioinformatics to preliminarily investigate the the mechanism of differential miRNA in the occurrence and development of Standford type B acute aortic dissection.MethodPart ⅠFrom July 2017 to May 2019,192 cases with phlegm-blood stasis type of Standford type B AAD were included according to the inclusion and exclusion criteria.They were divided into death group and survival group according to the survival status of patients during hospitalization.The baseline data,laboratory findings,treatment protocol,and clinical endpoints during hospitalization were enrolled.Univariate COX survival analysis was performed to detect the potential risk factors that affect the risk of death during hospitalization of patients with Standford Type B AAD.Multivariate COX survival analysis was performed based on statistically significant differences,and the independent risk factors for patients’in-hospital death with Standford Type B AAD were determined.The ROC curve was used for determining the diagnosis efficiency and best cut-off value of these independent factors.Kaplan-Meier survival analysis was used to determine the effect of independent risk factors on the cumulative survival rate of patients with Standford type B AAD during hospitalization.P<0.05 was considered statistically significant.SPSS25.0 software was used for statistical analysis.Part Ⅱ 5 patients with confirmed phlegm-blood stasis type of Standford Type B AAD were included,and matched one-to-one 5 healthy controls,collecting peripheral blood samples from patients and controls,and extracting total RNA from peripheral blood neutrophils.MiRNA microarray chips were used to detect the differential expression of miRNA in two groups of human neutrophils.The screening criteria for differential miRNA were that the expression level was fold change≥2 fold and P<0.05.The obtained differential genes are clustered and analyzed visually through heat maps,volcano maps.Bioinformatics approaches were used to predict the target genes of differential miRNA and the GO enrichment and signaling pathways involved.Results A total of 192 cases with phlegm-blood stasis type of Stand ford type B AAD were included in this study.28 patients died during hospitalization(14.58%).Six indicators inculding heart rate,white blood cell counts,neutrophil counts,total bilirubin,indirect bilirubin and uric acidin the death group were higher than those in the survival group.The significance was statistically significant(P<0.05),and the drug treatment ratewas higher in the death group(P<0.05).Univariate COX survival analysis shows that drug treatment,heart rate,white blood cell counts,neutroph ils,monocytes,alanine aminotransferase,uric acid were risk factors that increase the risk of death during hospitalization(P<0.05.Multivariate COX survival analysis showed that after adjusting for many confounding factors through stepwise regression,the white blood cell counts was an independent predictor of the risk of in-hospital death in patients with acute Standford Type B aortic dissection(HR:1.074,95%CI:1.019~1.133,P=0.008).Neutrophil counts as the major subtype of leukocytes also be an independent predictor of hospital mortality risk(HR:1.073,95%CI:1.018~1.132,P=0.009).The ROC curve shows that the white blood cell counts,neutrophil counts are highly effective in predicting the risk of death during hospitalization in patients with Standford Type B AAD.The area under curve(AUC)of white blood cell counts is 0.801(95%CI 0.687~0.914),the best predicted cut-off value is 15.8×10~9/L,the sensitivity is 0.615,and the specificity is 0.873.The AUC of neutrophils is 0.789(95%CI 0.682~0.897),the best predicted cut-off value is 9.52×10~9/L,sensitivity is 0.846,specificity is 0.678.Kaplan-Merier survival analysis showed that patients with white blood cell counts≥15.8×10~9/L at the time of admission had a lower cumulative survival rate during admission than patients with white blood cell counts<15.8×10~9/L(χ~2=18.117,P<0.001),while neutrophils≥9.52×10~9/L the cumulative survival rate was significantly reduced(χ~2=8.29.P=0.004).The miRNA chips analysis showed that there were 12 miRNA expression differences between the phlegm-blood stasis type of Standford Type B AAD group and control group(fold change>2.0,P<0.05),in which there were 3 miRNA up-regulated and 9 miRNA down-regulated in the AAD group,the highest up-regulation multiple was miR-629-3p,and the most down-regulated multiple was miR-598-3p.Target gene prediction shows that miR-629-3p and miR-598-3p participate in the regulation of many target genes.The signaling pathways analysis showed that differential miRNA may participate in the pathophysiologic mechanisms of AAD by regulating the VEGF signaling pathway and MAPK signaling pathway.Conclusions 1.neutrophil counts is an independent risk factor for hospital mortality in patients with phlegm-blood stasis type of Standford type B AAD.2.There were differential expression of miRNA in circulating neutrophils from patients with phlegm-blood stasis type of Standford type B AAD and controls,bioinformatics analysis suggests that differential miRNA in neutrophils may participate in the pathophysiologic mechanisms of AAD by regulating the VEGF signaling pathway and MAPK signaling pathway.