| Objective: 1.The research systematically analyzes the ancient and modern literature,studies the classification and identification system of body constitution in TCM,clarifies the connotation and extension of body constitution in TCM,and lays a theoretical foundation for its clinical application.2.In order to explore an effective and objective way of classifying and identifying constitution,the SNP sites of mitochondrial gene of kidney yang deficiency constitution were screened by mitochondrial genome sequencing method.The molecular biological basis for identifying kidney yang deficiency constitution was explored through site function annotation,and possible biomarkers were screened for classification and identification of kidney yang deficiency constitution.Methods: 1.Literature research(1)Literature retrieval: The key words of “Ti-zhi”,“Bin-zhi”,“Su-zhi”,“Bin-fu”,“Qi-bin”,“Bin-zhi” and “Su-bin” were searched in the large electronic series "Zhong Hua Yi Dian"(Chinese medical classics).In addition,the keywords of "constitution connotation","constitution identification","constitution diagnosis","constitution classification","constitution of traditional Chinese medicine" and "constitution of TCM" were searched in articles published from January 1979 to December 2018 on CNKI and Pub Med.(2)Literature inclusion and exclusion: According to the inclusion criteria and exclusion criteria.(3)Literature summary: The literatures were sorted out and summarized by systematically analyzing ancient and modern literature,carrying out in-depth research on TCM constitution classification and identification methods and exploring the extension of constititution theory,laying a foundation for clinical application of constitution in TCM.2.Study on SNP of mitochondrial DNA with kidney yang deficiency.(1)The subjects with kidney yang deficiency constitution were included: the subjects with kidney yang deficiency constitution were selected according to the evaluation standard of kidney yang deficiency constitution formulated by the research group in the early stage,and the rest of the subjects with yang deficiency constitution were excluded by the classification standard of constitution.After their diagnosis was confirmed by 3 professionals in TCM diagnostics,the subjects signed the informed consent and were included in the research.(2)Mitochondrial gene sequencing: After the blood samples were collected from the subjects with kidney yang deficiency constitution,the mitochondrial DNA was extracted,and the mitochondrial gene was sequenced after quality inspection and amplification.(3)Screening of mitochondrial gene SNP sites: Comparing the DNA sequence of the included subjects with the standard sequence of mitochondrial genes,the common SNP sites of subjects with kidney yang deficiency constitution were screened.(4)Annotation of molecular biological function of SNP site in mitochondrial gene.Results: 1.Literature research(1)There are 3997 entries in ancient literature retrieval and 2603 articles in Chinese and 150 articles in English in modern literature.According to the inclusion and exclusion criteria,365 Chinese and 16 English literatures were included.(2)Constitution formation: The constitution is an inborn quality which is further moulded after the person is born.(3)Constitution connotation: Constitution is the human body characteristic expressed in the aspects of spirit,emotion and physique by the joint action of congenital endowment and acquired environment under the guidance of the concept of “Xin Shen He Yi”(body-mind harmony).(4)Constitution classification: Ancient constitution classification is based on "yin and yang" and "five elements".The chapters of Yin Yang Er Shi Wu Ren(The Yin and Yang Categorization and the 25 Human Types)and Tong Tian(To Penetrate Heaven)from Ling Shu(Spiritual Pivot)have the most detailed record of classification and coverage.Modern constitution classification mainly includes pathological constitution classification and more diversified comprehensive classification,as well as constitution classification of special groups(such as children and women).(5)Constitution identification: Constitution in ancient Chinese classics is very rich in its connotations,mainly including physical characteristics,physiological characteristics and psychological characteristics.Constitution identification research in modern TCM focuses on the objectification of constitution,and TCM body mass table is most widely used.In addition,the objectification of the four diagnostic information and the introduction of modern biological indicators have promoted the development of constitution objectification.(6)Constitution classification and identification system framework: Formed a TCM constitution classification and identification system framework with the concept of "integration of body and mind" as the core,appearance characteristics as the basis,molecular biology and objective four diagnostic information data as reference,and physique questionnaire data as auxiliary.2.Study on SNP of mitochondrial gene in subjects with kidney yang deficiency(1)31 cases of kidney yang deficiency were included by body mass scale and professional evaluation.(2)There are 10 common mutation sites in the mitochondrial DNA of the subjects with constitution of kidney yang deficiency.MT-RNR1(m.73A>G,m.263A>G,m.750A>G,m.1438A>G),MT-RNR2(m.3105ACN>AC),MT-CO1(m.7028 C>T),MT-ATP6(m.8860A>G),MT-ND4(m.11719 G> A),MT-CYB(m.14766 C>T,m.15326A>G).Two of the loci,m.73A>G and m.263A>G,were located in the non-coding region upstream of mitochondrial DNA,and the rest were located in the exon region.(3)Gene locus annotation found that kidney yang deficiency constitution may be associated with aging process,energy metabolism,reproductive function and mental disorders.Conclusion: 1.Constitution is a kind of human body characteristic which can be manifested in spirit,emotion and body under the guidance of "Shen Xin He Yi"(body-mind harmony)which is influenced by the joint actions of innate endowment and acquired environment.The thought of body-mind harmony runs through the whole process of constitution classification and identification.It is the core idea of constitution theory.2.The constitution classification and identification system is a useful exploration for the objectification of constitution diagnosis,since it is taking the concept of "body-mind harmony" as the core,appearance characteristics as the basis,molecular biology and objective four diagnostic information data as reference,and constitution questionnaire data as auxiliary.3.Mitochondrial dysfunction may be the key link in the occurrence of kidney yang deficiency constitution,and the change of mitochondrial gene SNP may be the biological basis for the formation of kidney yang deficiency constitution,which provides a new research perspective for the study of the essence of kidney yang deficiency.4.The six genes and loci screened out in this study may be potential biomarkers for the identification of kidney yang deficiency constitution and the targets for regulating kidney yang deficiency constitution.5.The basic research on molecular biology of constitution may be one of the effective ways to realize the objectification of constitution classification and identification... |
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