Yang And Qualitative Theory, And Its Gene Expression Profiles In Peripheral Blood

Objective To construct a theoretical basis for Yang-qi deficiency constitution. To reveal the molecular mechanism for Yang-qi deficiency constitution by detecting the gene expression profiles of peripheral blood. Methods Discussions related with yang-quip deficiency constitution in more than 600 ancient documents and lots of modern literatures were searched and collected. The concept, cause of formation, characteristics, diagnosis and intervention method of yang-qi deficiency constitution were studied, then the theoretical basis of yang-qi deficiency constitution was constructed. Eight typical subjects of yang-qi deficiency constitution and 6 subjects of normal constitution were selected on the basis of clinical epidemiological investigation. Ten milliliters of fasting venous blood were collected from each subject. The white blood cells were isolated, and the total RNA was extracted, purified and amplified. The complement RNA (cRNA) was labelled with biotin. The gene expression profiles of peripheral blood were detected with U133 plus 2.0 gene chip of Affymetrix Corporation. The differentially expressed genes as compared with normal constitution were analyzed with software of Significance Analysis Microarray (SAM), and they were annotated and statistically analyzed in websites of GO (http://www.geneontology.org), KEGG (http://www.genome.ad.jp), BioCarta (http://www.bioCarta.com) and GenMAPP (http://www.genmapp.org). The biological functions of differentially expressed genes were evaluated in order to reveal the cause, characteristics and liability of yang-qi deficiency constitution at the level of gene. Results Yang-qi deficiency constitution is caused not only by congenital factors such as innate deficiency and improper nursing during pregnancy, but also acquired predispositions such as social environments, eating habits, diseases and doctors′mistakes. The characteristics of yang-qi deficiency constitution are cold appearances (creeping chill, cold and clear excretion and an aversion to raw and cold foods), obesity, whitish complexion and faint pulse, etc. Yang-qi deficiency constitution has a susceptibility to asthenia cold syndrome, cold-dampness syndrome, blood-stasis syndrome, phlegm retention, tumidness, diarrhea and impotence, etc. A quantized diagnosis of yang-qi deficiency constitution should be performed on the basis of retrieval studies and clinical experiences of experts. The treatment principles of yang-qi deficiency constitution include warming and invigorating yang-qi with drugs of sweet taste and warm property, warming and invigorating yang-qi of the gate of life with drugs of yang-warming and cold-dispelling. During the treatment, it should be noticed that yang-qi and yin-essence could reciprocally transformed into each other, so does the essence and qi. As the fold change was more than 2.0 or less than 0.5, there were 1739 probe sets of up-regulated expression in the group of yang-qi deficiency constitution as compared with normal constitution group. Of the 1739 probe sets, 785 were up-regulated and 957 down-regulated. As q < 0.05, 684 probe sets were up-regulated, and 920 down-regulated. Two hundreds and sixty-seven differentially expressed probe sets could be annotated and biologically analyzed in websites of GO, KEGG and GenMAPP. Of the 267 probe sets, 150 was up-regulated, 117 down-regulated. There were 48 biological functions in GO annotation, 3 pathways in KEGG analysis and 9 pathways in GenMAPP analysis. The up-regulated genes included inflammatory cytokines-related genes, CREB (cAMP response element binding protein) and CREM (cAMP response element binding module), etc. The down-regulated genes included genes related with genetic information transferring and MTHFR (5, 10-methylene tetrahydrofolate reductase), etc. Conclusion The theoretical system of yang-qi deficiency constitution could contribute to clinical medication and health preservation. The mechanism of yang-qi deficiency constitution is related with lots of disorders of physiological function. Its molecular mechanism is somewhat complicated, and should not be explored on the basis of several genes and their expression products. Its mechanism might associate with immunological disorders characterized by the up-regulation of inflammatory cytokines-related genes and the decrement of ability of genetic information transferring.