The Correlation Of ApoE Gene Polymorphism With TCM Syndromes Of Coronary Heart Disease And Its Common Risk Equivalent Diseases

Disease differentiation is the basic principle for Traditional Chinese Medicine(TCM)to recognize and treat diseases.Chinese medicine emphasizes treatment according to disease differentiation,and accurate differentiation can not only master the pathogenesis of disease,but also have an important effect on the progress of disease.In recent years,the objectivity of the clinical test index has laid the foundation for the disease differentiation and classification of TCM.By exploring the correlation between objective indexes and TCM diseases of CHD,we can intervene in the pathological evolution process as early as possible and better reflect the advantages of TCM in preventing and treating CHD.CHD equivalent disease is defined as one having the equivalent absolute risk of major cardiovascular events in patients without CHD within 10 years,compared to that of patients with CHD,type 2 diabetes mellitus and ischemic stroke are common diseases of CHD,and their damages to the human body and cardiovascular events caused serious harm to human health.Thus to improve the treatment level and take fully advantages of TCM in the treatment of type 2 diabetes mellitus and Ischemic Stroke is an important problem to be solved.Further understanding the regularity of TCM disease differentiation in patients with CHD and common equivalent clinical risk disease,exploring the correlation between different disease types of CHD and its common riskequivalent disease and modern objective examination indexes on the basis of combination of disease and disease,to provide the basis for the objectivity and standardization of TCM disease differentiation and classification,so as to better play to the characteristics and advantages of the intervention of Chinese medicine on CHD and its clinical common risk equivalent diseases.This paper is divided into two parts:literature review and clinical research.I Literature review:including the following two parts1.Research progress on lipoprotein and TCM disease differentiation of CHDpatientsIn recent years,many studies have confirmed that lipoprotein metabolism disorder,as one of the important pathogenesis of CHD,is closely related to the occurrence and development of CHD.ApoE gene plays an important role in lipoprotein metabolism,its gene polymorphism determines blood lipid level in individuals.This review summarizes the correlations between ApoE and CHD,lipoprotein and CHD disease differentiation.It can provide reference for the objectification of CHD disease differentiation.2.Research progress of TCM Diseases of CHD and its clinical common risk equivalent diseasesThe etiology and pathogenesis and differentiation of symptoms and signs for classification of disease of CHD and its clinical risk equivalent diseases were reviewed,involving classical theory of ancient books,clinical experience of modern doctors,national industry classification standards and modembiological indicators.Since the subjectivity of disease differentiation in clinical experience,the study of modern objective indexes is still in the preliminary stage,and most studies are small samples,the law of disease differentiation still needs to be studied and summarized.Ⅱ Clinical research:including the following two partsStudy Ⅰ:Relationship between ApoE gene polymorphism and TCM disease type of CHDObjective:Based on the theory of TCM disease differentiation,referring to the literature of past years,combining with modern molecular biology technology,we explored the genetic rule of CHD and analyzed the correlation between TCM disease type and polymorphism of ApoE gene in patients with CHD,thus to provide a basis for the objectification of TCM disease differentiation.Methods:917 patients with CHD from Xiyuan Hospital,Chinese Academy of Traditional Chinese Medicine from December 2016 to August 2017 were selected.According to the criteria of TCM Disease Differentiation of Coronary Heart Disease and the Clinical Guiding Principles of New Chinese Medicine,differentiation of symptoms and signs for classification of disease,2ml venous blood was extracted from the participants,polymorphism of ApoE gene was detected by Gene sequencing,analysis of the distribution of disease types of CHD,the distribution of ApoE gene and the correlation with TCM disease type were performed using statistical methods.Results:(1)Distribution of TCM diseases of CHD:disease of combination of phlegm and stasis is the most common in CHD,300 cases(32.7%of the total number of cases,applicable to the following),the second is Qi-deficiency and Blood Stasis Disease,158 cases(17.2%)and Qi-deficiency and Blood Stasis and Turbid Phlegm Disease,134 cases(14.6%),none of the remaining symptom types exceed 10%of the total number of cases,i.e.,Heart Pain with Disease of Yang Deficiency and Cold Congelation,29 cases(3.2%);Dual Deficiency of Qi and Yin,84 cases(9.2%);Dual Deficiency of Qi and Yin and Heart-blood Stasis Disease,66 cases(7.2%);Disease of Qi Stagnation and Blood Stasis,57 cases(6.2%);Disease of Yin Deficiency of Heart and Kidney,30 cases(3.3%);Heart-blood Stasis Disease,42 cases(4.6%);Edema due to Yang Insufficiency,17 cases(1.9%),respectively.(2)In 917 patients with CHD,there were three homozygous gene phenotypes E2/E2,E3/E3,E4/E4,and three heterozygote phenotypes,E2/E3,E2/E4,E4/E4,in which E3/E3 gene phenotypes are predominant,610 cases,up to 66.5%;the next are E2/E3 and E3/E4 phenotypes,111 cases(12.1%)and 154 cases(16.8%),respectively;E2/E2,E2/E4,E4/E4 gene phenotypes are the least,14 cases(1.5%),13 cases(1.4%)and 15 cases(1.6%),respectively.(3)Comparison of phenotype distribution of ApoE gene in different TCM diseases of CHD:the E3/E3 gene phenotype frequency of Deficiency of Qi and Yin and Heart-blood Stasis Diseaseis issignificantly lower than the Disease of Combination of Phlegm and Stasis,the difference is significant(P<0.01);the E3/E4 gene phenotype frequency of Deficiency of Qi and Yin and Heart-blood Stasis Diseaseis is significantly higher than the Disease of Combination of Phlegm and Stasis,the difference is statistically significant(P<0.05);while the comparison of other gene phenotypes in Deficiency of Qi and Yin and Heart-blood Stasis Diseaseand Disease of Combination of Phlegm and Stasis,the difference is not statistically significant(P>0.05).Comparison of any other TCM Disease types between two groups,the difference is not statistically significant(P>0.05).(4)Comparison of phenotype(combined)distribution of ApoE gene in different TCM diseases of CHD:referring to the literature domestically and abroad,the similar genetic phenotypes are combined to be analyzed.Similar genotypes are also combined in this paper and we found that the E2/E2+E2/E3 frequency of the Deficiency of Qi and Yin is significantly lower than the Edema due to Yang Insufficiency,the difference is statistically significant(P<0.05);comparison of the E2/E4+E3/E4+E4/E4 or E3/E3 of Deficiency of Qi and Yin and Edema due to Yang Insufficiency,however,the difference is not statistically significant(P>0.05).The E2/E4+E3/E4+E4/E4 frequency of the Deficiency of Qi and Yin and Heart-blood Stasis Diseaseis is significantly higher than Qi Stagnancy and Blood stasis,the difference is statistically significant(P<0.05);for comparison of the E2/E2+E2/E3 or E3/E3 of the Deficiency of Qi and Yin and Heart-blood Stasis Disease and Qi Stagnancy and Blood Stasis,the difference is not statistically significant(P>0.05).The E2/E4+E3/E4+E4/E4 frequency of the Deficiency of Qi and Yin and Heart-blood Stasis Diseaseis is significantly higher than the Disease of Combination of Phlegm and Stasis,and the difference is statistically significant(P<0.05);the E3/E3 frequency of the deficiency of qi and yin and heart-blood stasis disease is significantly lower than the disease of combination of phlegm and stasis,the difference is statistically significant(P<0.05);while comparison of the E2/E2+E2/E3 of the deficiency of qi and yin and heart-blood stasis disease and the disease of combination of phlegm and stasis,the difference is not statistically significant(P>0.05).(5)Comparison of phenotype distribution of ApoE gene in different TCM diseases(classified as asthenia disease,empirical disease,asthenia disease mingling with empirical disease)of CHD:empirical disease E3/E3 gene phenotype frequency is significantly higher than the asthenia disease mingling with empirical disease,the difference is statistically significant(P<0.05);empirical disease E3/E4 gene phenotype frequency is significantly lower than the asthenia disease mingling with empirical disease,the difference is statistically significant(P<0.05);empirical disease E2/E4+E3/E4+4/E4 phenotype frequency is significantly lower than the asthenia disease mingling with empirical disease,the difference is not statistically significant(P<0.05);for comparison of other gene phenotypes between empirical and asthenia disease mingling with empirical disease,the difference is statistically significant(P>0.05).Comparison between empirical and asthenia disease,and comparison between asthenia disease mingling with empirical disease and asthenia disease,the difference is not statistically significant(P>0.05).Conclusion:(1)In this paper,the main diseases of CHD are qi-deficiency and blood stasis,Qi deficiency and blood stasis and turbid phlegm disease,and disease of intermin-gled phlegm and blood stasis.(2)A total of E2/E2,E2/E3,E2/E4,E3/E3,E3/E4 and E4/E4 of the CHD ApoE gene phenotype are detected,E3/E3 is the main gene phenotype of each TCM disease type.(3)The ApoE gene phenotype is correlated with TCM diseases of CHD.(4)ApoE gene phenotype is correlated with the empirical and asthenia disease mingling with empirical disease of CHD.Study Ⅱ:The correlation between ApoE gene polymorphism and the common risk equivalent disease of CHDObjective:To observe the correlation between the polymorphism of ApoE gene and the common risk equivalent diseases of CHD,thus making the TCM disease differentiation and classification more standardized,improving the level of TCM Disease differentiation and treatment and clinical efficacy in patients with CHD and other risk equivalentdiseases,reducing the incidence of cardiovascular events.Methods:95 patients with type 2 diabetes mellitus and 208 patients with ischemic stroke from December 2016 to August 2017 were enrolled in our hospital,according to the standard of TCM disease differentiation.Type 2 diabetes mellitus is divided into brandy nose with pattern of heat exuberance in lung and stomach,liver-kidney yin deficiency disease,disease of deficiency of both qi and yin,disease of internal obstruction of phlegm turbidity,and deficiency disease of both Yin and Yang.Ischemic stroke is divided:into apoplexy with disease of upward disturbance of wind-fire,disease of blood stasis due to qi deficiency,phlegm-heat excess disease,phlegm-blood accumulation and disease of wind stirring due to yin deficiency.Gene phenotypic distribution of different TCM diseases in patients with these two diseases was detected and was analyzed by SPSS 22.0 statistical software.Results:1.Type 2 diabetes mellitus(1)In the 5 types.of TCM diseases of type 2 diabetes mellitus:Disease of internal obstruction of phlegm turbidity,44 cases(46.3%)is the most,the next is dual deficiency of qi and yin,22 cases(23.2%),pattern of heat exuberance in lung and stomach,14 cases(14.7%),disease of yin deficiency of liver and kidney,12 cases(12.6%),deficiency disease of both Yin and Yang,3cases(3.2%).(2)Two homozygous phenotypes of E2/E2,E3/E3 and two heterozygous phenotypes ofE2/E3,E3/E4 were existed in 95 diabetic patients.E3/E3 gene phenotype accounts for the majority,69 cases,the proportion is as high as 72.6%;the second is E2/E3 and E3/E4 gene phenotype,14 cases(14.7%)and 11 cases(11.6%);E2/E2 gene phenotype is the least,only 1 case(1.1%).(3)Comparison of phenotypic distribution of ApoE gene in different TCM Disease Types in type 2 diabetes mellitus:heat exuberance in lung and stomach type E3/E3 gene phenotype frequency is obviously lower than the type of yin deficiency of liver and kidney,the difference is highly statistically significant(P<0.01).Comparison of the other two groups between TCM Disease types,the difference is not statistically significant(P>0.05).(4)Combination analysis of genotypes:in addition to the heat exuberance in lung and stomach type and liver and kidney yin deficiency type comparison,for comparison between any other two groups of TCM diseases,the difference is not statistically significant(P>0.05).The E2/E2+E2/E3 frequency of the heat exuberance in lung and stomach type is obviously higher than the type of yin deficiency of liver and kidney,the difference is statistically significant(P<0.05);frequency of E3/E3 type in heat exuberance in lung and stomach type is obviously lower than the type of yin deficiency of liver and kidney,the difference is highly statistically significant(P<0.01);while comparison of the E3/E4 between Lung and stomach Heat Kidney Type and liver and Kidney Yin deficiency type,the difference is not statistically significant(P>0.05).(5)5 disease types of diabetes are classified as deficiency and empirical evidence:no matter whether the genotype was combined or not,there is no statistical significance between the empirical and the deficiency disease(>0.05).2.Ischemic stroke(1)Ischemic stroke patients are divided into the following.5 TCM disease types:apoplexy with disease of upward disturbance of wind-fire,41 cases(19.7%);disease of blood stasis due to qi deficiency,52 cases(25.0%);apoplexy with disease of excessivefu-viscera caused by phlegm-heat,3 cases(1.4%);disease of phlegm and stasis blocking collaterals,67 cases(32.2%);apoplexy with disease of wind stirring due to yin deficiency,45 cases(21.6%).(2)In 208 patients with ischemic stroke,there are 3 homozygous phenotypes of E2/E2,E3/E3,E4/E4,and three heterozygous phenotypes of E2/E3,E2/E4,E4/E4.E3/E3 gene phenotype accounts for the overwhelming majority,149 cases,up to 71.6%;the second is the E2/E3 and E3/E4 gene phenotypes,27 cases(13.0%)and 29 cases(13.9%);E2/E2,E2/E4,E4/E4 gene phenotype is the least,only one case(0.5%),respectively.(3)Comparison of phenotypic distribution of ApoE gene in different TCM Disease Types in ischemic stroke:the E2/E3 frequency of the apoplexy with disease of upward disturbance of wind-fire is significantlylower than the apoplexy with disease of blockade of wind-phlegm-static blood.the difference is statistically significant(P<0.05).The E3/E3 frequency qi deficiency and blood stasis type is significantly lower than the yin asthenia causing wind,the difference is statistically significant(P<0.05);the E3/E3 frequency of phlegm-stasis blocking collateral type is significantly lower than the yin asthenia causing wind type,the difference is statistically significant(P<0.01);the E3/E4 frequency of the phlegm-stasis blocking collateral type is apparently higher than the yin asthenia causing wind type,the difference is statistically significant(P<0.05);comparison of TCM Disease types between any other two groups,the difference was not statistically significant(P>0.05).(4)Combination analysis of genotypes:In addition to the comparison of Qi deficiency and blood stasis type and yin asthenia causing wind,phlegm and stasis obstruction of collaterals,comparison of TCM Disease types between any other two groups,the difference is not statistically significant(P>0.05).The E3/E3 frequency of Qi deficiency and blood stasis type is significantly lower than yin asthenia causing wind type,the difference is statistically significant(P<0.05);while for comparison of the E2/E2+E2/E3 or E2/E4+E3/E4+E4/E4 of Qi deficiency and blood stasis type,the difference is not statistically significant(P>0.05).The E2/E4+E3/E4+E4/E4 frequency of phlegm and stasis blocking collaterals type is significantly higher than yin asthenia causing wind type,the difference is statistically significant(P<0.05);the E3/E3 frequency of phlegm and stasis blocking collaterals type is significantly lower than yin asthenia causing wind type,the difference is statistically significant(P<0.01);while for comparison of the E2/E2+E2/E3 between phlegm and stasis blocking collaterals type and yin asthenia causing wind type,the difference was not statistically significant(P>0.05).(5)5 disease types of Type 2 diabetes mellitus are classified as asthenia disease,empirical disease,and asthenia disease mingling with empirical disease:for comparison of the asthenia disease and empirical disease,the empirical E3/E3 gene phenotype frequency is obviously lower than asthenia disease,the difference is highly statistically significant(P<0.01);the empirical E3/E4 gene phenotype frequency is obviously higher than asthenia disease,the difference is highly statistically significant(P<0.05);for comparison of other gene phenotypes between asthenia disease and empirical disease,the difference is not statistically significant(P>0.05).For comparison of the asthenia disease and asthenia disease mingling with empirical disease,asthenia disease mingling with empirical disease E3/E3 gene phenotype frequencyis obviously lower than asthenia disease,the difference is statistically significant(P<0.05);for comparison of other gene phenotypes between deficiency disease and deficiency disease,the difference is not statistically significant(P>0.05).For comparison of the asthenia disease and empirical disease mingling with empirical disease,the difference is not statistically significant(P>0.05).Conclusion:(1)In this paper,the common disease of type 2 diabetes mellitus is the disease of internal obstruction of phlegm turbidity;the distribution of apoplexy with disease of excessive fu-viscera caused by phlegm-heat in ischemic stroke is little,however,the distribution of other diseases is uniform.(2)A total of 4 gene phenotypes are detected in type 2 diabetes mellitus ApoE,including E2/E2,E2/E3,E3/E3,E3/E4;a total of 6 gene phenotypes are detected in ischemic stroke,including E2/E2,E2/E3,E2/E4,E3/E3,E3/E4,E4/E4,both mainly dominated by E3/E3.(3)ApoE gene phenotype has certain correlation with TCM diseases of type 2 diabetes mellitus and ischemic stroke.(4)ApoE gene phenotype has no correlation with type 2 diabetes mellitus empirical disease,asthenia disease,and asthenia disease mingling with empirical disease;while it has some correlation with ischemic stroke empirical disease and asthenia disease,and asthenia disease,asthenia disease mingling with empirical disease.