| Stroke is one of the most important reasons of death,and the incidence of ischemic stroke is roughly from 60% to 80%,which brings heavy burden to the society and family.Therefore,to investigate the risk factors and the biomarkers of stroke is of great importance,which can be used to screen the highly-risk individual and reduce the incidence of stroke.Part 1 Comparison analysis among the methods of VASP,PL-11 and ADP-induced flow cytometryAim: This part compared the methods of VASP,PL-11,and ADP-induced flow cytometry,aiming to find the most convenient and cost-efficient method to measure the platelet aggregation rate.Materials and methods: 43 patients with ischemic stroke were enrolled from Jan.1 to Aug.1 in Tianjin Medical University in 2015.The platelet aggregation rate was respectively measured by VASP,PL-11,and ADP-induced flow cytometry.Pearson analysis was used to analyze the correlation,and Bland-Altman analysis was used to analyze the consistence.The NIHSS score and the mi R-223 level were also measured.Results:1.The average values corresponding to VASP,PL-11,and ADP-induced flow cytometry were 59.46%,60.39%,and 50.23%,respectively.The results show that the Pearson coefficient between VASP and PL-11 is 0.901(p=0.01),and the Pearson coefficient between between VASP and PAg-ADP is 0.843(p=0.02).Both the results of Bland-Altman analysis show a good consistency.VASP-PRI was negatively correlated with mi RNA-2233,and positively correlated with NIHSS score.2.The Pearson coefficient between VASP and PL-11 is 0.901(p=0.01),3.The Pearson coefficient between between VASP and PAg-ADP is 0.843(p=0.02).Both the results of Bland-Altman analysis show a good consistency.4.The platelet aggregation rate measured by VASP-PRI was reduced after using clopidogrel.5.The homocysteine level was positively correlated with platelet aggregation rate.6.VASP-PRI was negatively correlated with mi RNA-223(r=-0.519,p=0.02).7.VASP-PRI was positively correlated with NIHSS score(r=0.424,p=0.03).8.Patients with progressive stroke had a higher platelet response than those with non-progressive stroke(VASP:59.4947±9.1300 vs 38.6585±21.3911,p=0.0001,PL-11:62.7316 ±10.8917 vs 39.4415 ±14.9789,p <0.0001,PAg-ADP 63.6263 ±7.0973 vs 50.6200±15.9416,p=0.0010).Fasting blood glucose within 48 hours of onset was higher in progressive stroke than that in non-progressive stroke(8.1306±2.9253 vs 6.4330±2.0155,p=0.0109).Conclusion:1.The results of VASP,PL-11,and PAg-ADP show a good consistency.2.Clopidogrel can reduce the platelet reactivity.3.The blood homocysteine level may be associated with the platelet reactivity.4.The mi RNA-223 may be related to the platelet reactivity.A higher level of mi RNA-223 could lead to a higher level of platelet reactivity.5.The platelet reactivity may be related to the severity of ischemic stroke.6.Patients with progressive stroke have elevated platelet reactivity and the hyperglycemia may be the risk of progressive stroke.Part 2 Metabolomics study of ischemic stroke patientsAim: Ischemic stroke is an important cause of death,which can severely affect the quality of life and bring the heavy burden to the society and family.To investigate the effective treatment of ischemic stroke is of great importance.This part aims to investigate the changes of plasma metabolites in the patients with ischemic stroke,and to analyze the metabolism pathways.Metarials and methods: 47 patients with ischemic stroke and 30 healthy controls were enrolled.The plasma samples were analyzed by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-QTOF-MS)to create the metabolomics profiles.Principal component analysis and partial least squares discriminate analysis were used to investigate the metabolic changes related to ischemic stroke.The receiver operator characteristic curves were utilized to evaluate the specificity and the sensitivity of the obtained biomarkers.The metabolic pathways of the biomarkers were analyzed.Results:1.There was a significant difference in the peak intensity of the plasma in the healthy control group and the patients with acute ischemic stroke,indicating that the metabolites in the plasma were well separated.2.Eight biomarkers for ischemic stroke were obtained,including sphinganine,2-ketobutyric acid,tetradecanedioic acid,docosatrienoic acid,glutamine,phytosphingosine,lyso PE(0:0/22:0),and pyroglutamic acid.3.The results of the metabolism analysis show that sphingolipid metabolism,D-glutamine and D-glutamate metabolism,alanine,aspartate and glutamate metabolism,valine,leucine and isoleucine biosynthesis,propanoate metabolism,and glutathione metabolism were perturbed.In addition,energy metabolism was also disturbed.4.Elevated sphingosine levels and a decrease in glutamine level suggest an increase in platelet reactivity.Conclusion: By using UPLC-QTOF-MS based metabolomics,our study shows that the metabolism has changed in the patients with ischemic stroke.The results could provide helpful targets for the treatment of ischemic stroke.Elevated sphingosine levels and a decrease in glutamine level suggest an increase in platelet reactivity. |
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