Exploring The Distribution Characteristics Of Ischemic Stroke Syndromes,clinical Risk Indicators And Their Association With Polk Polymorphism From The Perspective Of "same Disease With Different Syndromes"

Objective:This study aims to explore the distribution characteristics of ischemic stroke（IS）syndromes from the perspective of"same disease with different syndromes"in Chinese medicine,and use modern molecular biology techniques to analyze the essence of IS syndromes and the"same disease with different syndromes"The scientific connotation was explored,and the rs5744724 polymorphism of DNA polymerase kappa（POLK）gene was used as the core site to explore its association with IS TCM syndrome distribution and clinical key quantitative traits.Methods:A total of 774 ischemic stroke patients and 793 normal controls who met the enrollment criteria were collected.Diagnosis and classification of the case group,collect clinical information,including basic information,medical history,past history,physical examination,diagnosis,serological indicators（including lipid biochemistry,coagulation function,homocysteine,C-reactive protein,etc.）Serological indicators),standard syndrome scores,and establish a disease database with Excel software and Epidata software.Extract the whole genome DNA of the research object,use the Mass ARRAY SNP genotyping experiment technology of Agena platform to detect the polymorphic locus of the target gene,and use the specified version of SPSS software and PLINK and Editplus calculation software for statistical analysis of this study.Results:1.Distribution and characteristics of syndromes of"same disease with different syndromes"in patients with ischemic strokeThe mean age of ischemic stroke patients was（64.12±10.34）and the mean age of the control group was（63.81±10.36）.There was no statistically significant difference in sex composition between the two groups（χ2=2.815,P=0.093）,and there was no statistically significant difference in the mean age distribution between the two groups（t=-0.582,P=0.561）.The score of each syndrome was as follows:Wind syndrome was 9.53±2.75;The score of phlegm syndrome was9.99±2.512.The score of blood stasis syndrome was 8.63±1.727.The score of qi deficiency syndrome was 7.94±1.612.The score of fever syndrome was 8.8±1.907;The score of Yin deficiency and Yang hyperactivity syndrome was 11.17±3.666;In this study,338 cases of phlegm syndrome,307 cases of wind syndrome,171 cases of blood stasis syndrome,86 cases of heat,88 cases of qi deficiency syndrome,and 17 cases of Yin deficiency and Yang hyperactivity syndrome were obtained.2.Hardy-Weinberg Equilibrium（HWE）and genotype frequency distributionThe distribution of the POLK gene rs5744724 polymorphism locus in the control group was consistent with the Hwin equilibrium genetic law(P_HWE=0.053).There was no significant difference in rs5744724genotype frequency distribution between ischemic stroke syndrome group and control group（P>0.05）.Results The chi-square test results showed that the genotype frequency distribution of POLK gene rs5744724 site was significantly different between IS qi deficiency syndrome+IS blood stasis syndrome group and the control group（χ2=6.99,P=0.03）.3.Association analysis of the risk of ischemic stroke and POLK gene rs5744724 polymorphismThere was no significant correlation between rs5744724polymorphism in POLK gene and the risk of ischemic stroke in additive model（CC vs.CG vs.GG）,dominant model（CC+CG vs.GG）,recessive model（CC vs.CG+GG）and allele（C vs.G）（all P>0.05）.4.Association analysis of the risk of various symptoms of ischemic stroke and POLK gene rs5744724 polymorphism（1）in various genetic models,ischemic stroke,blood stasis[additive model:P=0.03;dominant model:P=0.04;allelic model:P=0.03),qi deficiency syndrome[dominant model:P=0.04]the happening of the risk and rs5744724 POLK gene polymorphisms were statistically significant,the correlation of combination of qi deficiency and blood stasis qi deficiency analysis found that rs5744724 POLK gene polymorphisms with ischemic stroke card+the happening of the risk of blood stasis syndrome significantly correlation[additive model:P=0.02;dominant model:P=0.01;allelic model:P=0.01],and there was no correlation between wind syndrome,heat syndrome,phlegm syndrome and POLK gene polymorphism in any of the genetic models.（2）after adjustment for age,gender stratification,ischemic stroke,blood stasis[additive model:P_adj=0.02;dominant model:P_adj=0.03],qi deficiency syndrome[dominant model:P_adj=0.04],qi deficiency syndrome and blood stasis[additive model:P_adj=0.02;dominant model:P_adj=0.01]risk associated with rs5744724 POLK gene polymorphism were significantly,but after adjustment for age and gender,in the genetic model,still did not see the rest of the three certificates（wind syndrome,heat,phlegm syndrome）statistical associations with POLK gene polymorphism.5.Association analysis of IS blood stasis syndrome,Qi deficiency syndrome score and POLK gene rs5744724 polymorphismThere was no significant difference between the score of blood stasis syndrome and Qi deficiency syndrome and the polymorphism of POLK gene rs5744724[P=0.77 for additive blood stasis syndrome,P=0.82 for dominant and P=0.76 for recessive;P=0.98 for additive qi deficiency syndrome,P=0.92 for dominant and P=0.75].After adjusting for gender and age indexes,there was still no significant statistical difference between the scores of the two synonyms（blood stasis syndrome and qi deficiency syndrome）and POLK polymorphism[blood stasis syndrome additive P_adj=0.79,dominant P_adj=0.86,recessive P_adj=0.76;qi deficiency syndrome additive P_adj=0.90,dominant P_adj=0.99,recessive P_adj=0.74].6.Correlation analysis of POLK gene polymorphism and blood pressure（systolic and diastolic）levels in patients with ischemic strokeThere was a statistically significant correlation between rs5744724polymorphism and systolic blood pressure in patients with ischemic stroke[additive model:P_adj=0.04].No statistical association was found between diastolic blood pressure level and POLK gene rs5744724polymorphism[additive model:P_adj=0.23;dominant model:P_adj=0.45;recessive model:P_adj=0.16].7.Correlation analysis of POLK gene polymorphism with fasting and postprandial blood glucose levels in patients with ischemic strokeRs5744724 POLK gene polymorphism with the object of study of fasting blood glucose levels and postprandial 2 hours blood glucose levels are not exist statistical correlation[fasting glucose additive model:P=0.69,the dominant model:P=0.35;implicit model:P=0.30;additive model:P_adj=0.74,the dominant model:P_adj=0.40;implicit model:P_adj=0.31;blood sugar 2 hours after meal additive model before correction:P=0.61,the dominant model:P=0.34;implicit model:P=0.31;additive model:P_adj=0.68,dominant model:P_adj=0.37;recessive model:P_adj=0.24;].8.Analysis of the association between POLK gene polymorphism and coagulation markers in patients with ischemic strokeThere was no significant correlation between POLK rs5744724polymorphism and coagulation markers APTT,D-D,FIB,INR,PLT,PT,PTA and TT in patients with ischemic stroke.After adjusting for gender and age,no significant association was found between the polymorphic loci and coagulation markers.9.Correlation analysis between POLK gene polymorphism and blood lipid in patients with ischemic strokeSimilarly,before adjusting for gender and age,no statistical significance was found between POLK rs5744724 polymorphism and lipidemic indexes such as Apo-A,Apo-B,HDL,LDL,VLDL,TC,and TG in patients with ischemic stroke.Adjusting for gender and age factors,no statistical association was found between rs5744724polymorphism and all kinds of blood lipids in ischemic stroke patients.10.Correlation analysis of POLK gene polymorphism with Hcy and CRP in patients with ischemic strokeBefore adjusting for age and sex,no statistical association was found between C-reactive protein,homocysteine and POLK rs5744724polymorphism in ischemic stroke patients.After adjusting for age and sex,no statistical association was found between CRP and Hcy levels and POLK gene rs5744724 polymorphism.Conclusion:（1）According to the order of frequency,there were 338 cases,307cases,171 cases,86 cases,88 cases and 17 cases of syndromes classified by the Standard.The number of syndromes were phlegm,wind,blood stasis,qi deficiency,heat,Yin deficiency and Yang hyperactivity,and the former three were the most common.（2）Polymorphism at rs5744724 of the POLK gene may affect the occurrence of qi deficiency syndrome and blood stasis syndrome of IS,and may be an independent risk site for these two syndromes,but it does not serve as a risk genetic factor for other syndromes of IS.（3）The polymorphism of POLK gene rs5744724 was statistically significantly different from the level of systolic blood pressure in IS patients,and this SNP may also affect the level of systolic blood pressure in IS patients.However,in the correlation analysis of risk indicators such as blood lipid level,blood coagulation marker level,blood glucose level,CRP and Hcy,this study did not find a significant statistical correlation between POLK gene rs5744724 polymorphism and the above indicators in IS patients.