The Epidemiologic Investigation Of Sub-health State And Saliva Metabolomics Study Of Deficiency Of Its Spleen-QI Syndrome

BACKGROUNDSub-health state is the critical state of between health and sickness, and is some functionality change without disease. It could be incapable of accurate diagnosis for sickness according to its complicate clinical manifestation. World health organization (WHO) lately declaratory a predictive investigation disclosed that there are 75% sub-health population, only 5% healthy person all over the world, and the sub-health will be the formidable enemy of human health in 21 century. In late years, inland have carried out sub-health's study in succession, and gained some grade. Yet, a little key question do not solve, such as:①how many is sub-health's incidence?②what is the certain cause of sub-health ?③what is the sub-health's clinical manifestation ?④how is the sub-health intervened ?Being short of clear physical and chemical evidence of a disease, sub-health status can not be called "some disease". It is actually a syndrome to show that something wrong with physiology activities of the body. It is the early phase reporting the abnormality of the body. Taking concrete configuration as the basis, simple biological disease as the object of study, digitized checkout data as the diagnosing law, it is therefore very difficult to grasp the rule of diagnosing and treating sub-health status according to modern medicine. Traditional Chinese Medicine is one theory system which regards the human body itself as a whole and determines the treatment based in pathogenesis obtained through differentiation of symptoms and signs. Its main character different from the modern medicine is that it gives an important role to physiological character and functional rule of the body. Thus, TCM has a comprehensive advantage on knowing the essence of sub-health status and the way of diagnosis and treatment. The basis and characteristic essential of TCM is determination of treatment based in pathogenesis obtained through differentiation of symptoms and signs. Syndrome is the basis and premise of differentiation of symptoms and signs. It is also the key and bridge of connecting theory of TCM and system of clinical diagnosis and treatment. But the syndrome of Chinese medicine has obvious medical characteristic of experience. The research approach of the present stage adopts more the sorting and proving methods. Though it has made the diagnostic criteria of some syndromes, it is difficult to popularize extensively. Because these standards are established by experts' personal experience. In addition these syndromes often follow with the relevant disease. Different diseases have important influence to the syndrome. It is difficult to distinguish the peculiar symptom of syndrome and the symptom that take place due to the influence of disease. For example, there is difficult to adopt the same standard of the splenasthenic syndrome of digestive system and gynaecology. These questions have been all the time perplexing the standardization study of syndrome. The syndrome of sub-health status can get rid of the influence of disease. It will help to explore the inherent symptom characteristic of the syndrome, offer scientific basis for standardization study of syndrome, and promote the development of Chinese medicine. So, it is important scientific sense of the syndrome study of sub-health status.The spleen and stomach theory is important components in the theoretical system of Chinese medicine. Traditional Chinese medicine thinks that the spleen and stomach are the foundation of acquired constitution, the water and cereals sea, the source of generating QI and blood. The deficiency of spleen-QI is one of the most frequently syndrome of many kinds of diseases and sub-health state. So, it is very necessary to launch the modern research of the deficiency of spleen-QI. At present, though there are some study of the characteristic of deficiency of spleen-QI of department of internal medicine, pediatrics, gynaecology, etc., but do not discover the study of syndrome of sub-health. The "spleen controlling saliva" is one of the classical theories of Chinese medicine. It is experience of the long-term clinical practice and summary of the law, would be the certain scientific connotation. There are some the related research between the syndrome of spleen and the saliva at present. The saliva amylase has already consulted one of the standards as the diagnosis of deficiency of spleen-QI syndrome. However, there are some questions of the study, such as measuring in the simple biochemistry, relatively single of the research contents, low-levelly repeating, not enough of study depth, lack many indexes of monitor and study in step. So, it could be unable to announce true essence of the saliva changes of deficiency of spleen-QI. It must combine traditional theory of Chinese medicine with modern life science to introduce the new technology and methods to solve the problem of above-mentioned bottlenecks.Traditional Chinese medicine thinks that spleen control digestion, is the source of generating QI and blood and supply the nutrition to congenital essential substance to promote the growth of the human body, maintains the human life activity. Then the deficiency of spleen-QI must influence the function activity of many internal organs and substance metabolism to cause the metabolite change of the saliva. Modern life science think that the metabolite is in the end that the life adjusts and controls, so it approaches the biological phenotype even more than the genes and proteins. Then it could find the marker of the deficiency of spleen-QI syndrome from the saliva metabolite. It needs more overall and more systematic information in order to appraise the metabolism state of the organism, and this is exactly the problem that metabolomics can solve. Metabolomics is a science of the study of metabolite profiles of the organism in life activity process, is important component of systems biology. Genomics and proteomics usually find and distinguish the potential change, but metabolomics finds and distinguishs the true change. Just as Billy David said that the genomics and proteomics tell you what may take place, but the metabolomics tells you what has already happened. With development of the nuclear magnetic resonance (NMR), high-efficient liquid phase chromatogram (HPLC), mass spectrum (MS), bioinformatics, computer technology and so on, the technological platform of metabolomics become better day by day.OBJECTIVEFirst of all, we could study the emergence state, the clinical manifestation, the relevant influence factor, the distribution of the Chinese medicine syndromes and the characteristic of deficiency of spleen-QI syndrome through epidemiological investigation; Secondly, using the metabolomics research method, we could study the deficiency of spleen-QI salivary metabolite composition and variation in order to establish the foundation of the objective research for the deficiency of spleen-QI syndrome.METHODS(1) Use the approach of cross-sectional study, adopt the on-the-spot investigation method, let respondents fill in the sub-healthy questionnaire, combine the clinical examination results, judge the health status (include health, sub-health and disease). Gather the respondent's information material of the four diagnosis methods of the traditional Chinese medicine through doctor's interview at the same time and carry on differentiation of symptoms and signs according to the relevant syndromes standard of the Chinese medicine. Then, collect the information of the qualified questionnaires and enter Epidata3.02 database, use SPSS11.5 software to carry on descriptive statistics, Chi-Square test, Logistic regression analysis, factor analysis, etc. to study the emergence state, clinical manifestation, relevant influence factor, the distribution of the Chinese medicine syndromes and the characteristic of deficiency of spleen-QI syndrome.(2) Choose the sub-healthy deficiency of spleen-QI syndrome to be an experiment group, normal person as the normally contrast group and sub-healthy deficiency of kidney-YIN syndrome as the experiment contrast group, use pH appearance to determine pH value of the saliva, use the full-automatic biochemical analysis instrument to determine the biochemical index in the saliva, use the nuclear magnetic resonance (NMR) to determine 1H NMR of the saliva, centre its total mark value to convert with the proportion and carry on the principal component analysis (PCA) with Simca-P 10.0 software, in order to study the saliva metabolite profile of the sub-healthy deficiency of spleen-QI syndrome.RESULTS(1) In this investigation among 6110 people who come from the specific crowds in enterprises and medical university, there are 3981 people in sub-healthy state. The prevalence rate is 65.1%. The female is more than the male (P＜0.01). Most of the sub-healthy people are under 30 years old(account for 79.4%). As the age risen, the prevalence rate reduce (P＜0.01).(2) This analytical results of this investigation discovered that the 15 factors correlating with sub-healthy state are academic career, occupation, overtim, physical training, recreation activity, work ability, frustration, profile, income, inhabitation, interpersonal relationship, living environment, sense of safety etc(P＜0.05). According to B value, the academic career, overtime and frustration are the risk factors which induce the sub-health. Other 12 items such as occupation, physical training, recreation activity, physical training, recreation activity, work ability, profile, inhabitation satisfaction, income, interpersonal relationship, health status, living environment, sense of safety are the protection factors for sub-health.(3) This investigation discovered that the clinical manifestation of the sub-healthy state is very complicated, involve a lot of symptoms, such as physiology, psychology and society, etc. Use the factor to analyze, extract 10 factors altogether, combine medical knowledge further, can divide the clinical manifestation of the sub-healthy state into the society type (factor 6, show mainly as the drop of the communication frequency, burdensome corresponding social role, etc.), psychology type (factor 1, show mainly as nervous, unstable mood, anxious, depression, etc.), fatigue type (factor 3, 4, 9, show mainly as fatigue, debilitation, deficient breath, idle to speak, headache, dizzy, chest distress, short breath, flustered, abdominal distension, abdominal pain, anepithymia, aching pain in the waist and the back, leg knee getting aching and limp, etc.), astringent eye and dry pharynx type(factor 7, show mainly as astringent eye, sour and bloated eyes, mouth bitter, dry pharynx), urination and defecation dysfunction type (factor2, show mainly as stool dry and hard, stool thin, urinate netly, night urine grade frequently), irregular menstruation type (factor5, shown as dysmenorrhoea, dysfunction menstruation, leukorrhea increase ,etc.) and the physique type (factor 8, 10, shown mainly as easy to catch cold, aversion to wind, easy to diaphoresis, the hot center of the hand and the foot, etc.). There are seven subtypes totally. (4) This investigation discovered that the Chinese medicine syndromes of the subhealthy state involves 33, the first 10 syndromes are frequently, its cumulative percentage is up to about 70%. They are deficiency of spleen-QI, stagnation of liver-QI with deficiency of the spleen, hepatic and renal yin deficiency, deficiency of kidney-YIN, stagnation of QI due to depression of the liver, deficiency of kidney-YANG, deficiency of kidney-essence, deficiency of both heart and spleen, asdthenic splenonephro-yang, deficiency of liver-YIN, etc.(5) This investigation discovered that the main clinical manifestation of deficiency of spleen-QI of sub-health state are tired and weak, deficient breath and idle to speak, anepithymia, stool thin, easy to catch cold, easy to diaphoresis, abdominal distension, abdominal pain, aversion to wind, reaction dull, etc. Use the factor to analyze, the characteristic of deficiency of spleen-QI is:①the deficiency of vital energy behaves (show mainly as tired and weak, deficient breath and idle to speak, etc.);②spleen and stomach symptom (show mainly as anepithymia, abdominal distension, abdominal pain, stool thin, etc.);③physique state(show mainly as easy to catch cold, easy to diaphoresis, aversion to wind, etc.);④the pale tongue and the feeble pulses.(6) Compare with the control group, the pH value and the content of potassium (K), sodium(Na), chlorine(Cl), calcium(Ca), phosphorus(P), glutamic pyruvic transaminase(ALT), total albumen(TP), urea nitrogen(BUN), creatinine(Crea), uric acid(UA), lactate dehydrogenase(LDH), phosphocreatine kinase(CK), glucose(GLU) and triglycerides(TG) are not statistical significance in saliva of deficiency of spleen-QI(P＞0.05). However, the content of magnesium(Mg) and glutamic oxalacetic transaminase(AST) are obviously higher than control group (P＜0.05). In the saliva of deficiency of kidney-YIN, the content of P, Mg, AST, TG, BUN are obviously higher than control group (P＜0.05), and others is not statistical significance (P＞0.05). However, the content of TG in saliva of deficiency of kidney-YIN is higher than deficiency of spleen-QI group (P＜0.05).(7) The principal component value of saliva ~1H NMR of experiment every group distributes on three without overlapping and crossing regions in the ellipse scatterplot. The deficiency of kidney-YIN group concentrates on the left region, the deficiency of spleen-QI group concentrates on the right upper region, and the control group concentrates on the right bottom region. Compared with control group, lactic acid(1.34ppm), N-acetyl glycoprotein(2.1ppm), ethanol(1.18ppm) and glucose(3ppm-4ppm), etc, are relatively high in saliva of deficiency of spleen-QI, and acetate(1.9ppm, 1.94ppm), propionate(1.06ppm), alanine(3.78ppm),etc, are relatively high in saliva of deficiency of kidney-YIN.CONCLUSION(1) In the enterprise and medicine university, the prevalence of sub-healthy state is relatively high, it is 65.1%. It is involved in a lot of factors, such as individual diathesis, psychological state, work and life style, social environment, etc.(2) The clinical manifestation of the sub-healthy state is complicate with heterogeneity, and exist many kinds of clinical subtypes.(3) This investigation indicates that the Chinese medicine syndrome of sub-healthy state distributed fairly extensive. The deficiency of spleen-QI is one of common syndrome of sub-health, its main clinical manifestation characteristic is:①the deficiency of vital energy behaves;②spleen and stomach symptom;③physique state;④the pale tongue and the feeble pulses.(4) The saliva metabolite profiles of the deficiency of spleen-QI of sub-health obviously deviates from the normal crowd. It reveals the scientific characteristic of the theory of "the spleen controlling saliva".