The Research Of Asthenia Cold Syndrome On Symptomatic Rules And Its Metabonomics

Objective:1 Through the literature research, we wanted to summarize the cognition of ancient and modern researcher in etiological factor, type of syndrome and symptoms of asthenia cold syndrome, analyze the essence of asthenia cold syndrome. Thereby, we hoped to reveal the research's status quo of etiological factor, type of syndrome, symptoms and essence of asthenia cold syndrome. At the same time, we planned to combine with opinions of experts for formulating suitable clinical epidemiology investigational questionnaire of asthenia cold syndrome. And offered theoretical and literary foundation for the next study of etiological factor and symptomatic rule of asthenia cold syndrome.2 Through the clinical epidemiology investigating, we planned to get the case rate of asthenia cold syndrome by statistics and analyze the characteristic and rule of etiological factor, type of syndrome and symptoms of asthenia cold syndrome. Accordingly, we could understand the condition of what asthenia cold syndrome had taken place and look for its main Risk factor, as well as approach the distributive condition and the main indexes of differentiation of symptoms and signs of its type of syndrome. Moreover, we wanted to offer an example for the operational method of large sample clinical epidemiology research in symptoms of traditional Chinese medicine.3 We decided that we used the technique of nuclear magnetic resonance to approach changes of metabolin of asthenia cold syndrome and explain biological essence of asthenia cold syndrome. Then we hoped to explore the application of metabonomics'technology in the study of symptomatic essence.Method:1 The literature research Using yiji dictionary of china as retrieval instrument, we retrieved all of ancient times literature from Neijing term to Qing dynasty end and to arrange and analyze them; At the same time,we used china periodical full-text data base(library CNKI retrieval page of mdition university of heibei ) to retrieve modern literature which related with asthenia cold syndrome from 1996 to 2006, then carried out frequency analysis on symptoms and arranged them.2 The research of symptomatic rule of asthenia cold syndromeOn the literature's research and expert's consulation foundation, we formulated the investigational questionnaire of asthenia cold syndrome. After through the experts of Chinese traditional medicine, measuring scale, statistics, sociology demonstrasting, we beforehand investigate with investigational questionnaire. After amending, we appraised its reliability and validity by investigated again. Eventually, there formed a suitable investigational questionnaire of syndrome and etiology of asthenia cold syndrome, which possessed the qualitative of measuring scale. Used investigational questionnaire above, we adopted random cluster sampling, selected valid researcher who had been trained, then let them to survey 2500 inhabitant, who lived in shijiazhuang and its surrounding area in long-term. After these investigation questionnaires were reclaimed, they inspected and diagnosed by experts again and again. Among them, investigational questionnaires, which fit requirement, were carried out data record. Then we excavated and analyzed these data by multielement statistical analysis(cluster analysis, factor analysis, principal component analysis, logistic regression analysis) and theory of traditional Chinese medicine.3 The metabonomics'studyWe selected typical patients of asthenia cold syndrome menopathy as subjects, and selected typical patients of asthenia pyrosyndrome menopathy and health female as contrast. Every subjects took venous blood 5ml on an empty stomach at the second day of menstrual period. After centrifuging these venous blood, we took clear supernatant liquid 2ml and preserved it in - 70℃low temperature box, waiting for detection. We utilized 600MHz nuclear magnetic resonance spectrometer to detect serum metabolin composition. Then we adopted dalte software(http: //qobrue. usc. es) to get out data from subjects'serum 1HNMR spectra according toδ0.04 interval fromδ0 toδ6 segmentation. Except forδ4.669 which was water, we integrated others. After the integral of peak area had been carried out normalization, experts identified metabolin from every segment on the basis of literatures. Then according to location of peak and it representing metabolin's integrity, we gained 24 metabolin(including 2 unkown metabolin)and 1segment of glucose and amino acids. Through added up different segment of each metabolin, we got 25 score. The scores were Stored ExceL document, waiting statistical analysis.The remain serum was used to detect FSH,LH,E2,T and P by radio immunoassay.Result:1 The literature research1.1 The result of literature research shew that the cognition of etiological factor of asthenia cold syndrome mainly accord with ancient literature now; The differentiation of symptoms and signs of asthenia cold syndrome had not unified specification,and the cognition still wasn't unification; Forthcoming various kinds differentiation of symptoms and signs methods had directional significance. But these methods mostly set up on clinical report and individual empirical foundation, but were short of research of syndromes; On the clinic and scientific research, specialists thought highly of essence of syndromes but neglected content of syndromes themself.1.2 Through frequency statistics about symptoms of asthenia cold syndrome which had been described by modern literatures, the result shew that limbs cold, chilly, loose stool, lassitude had important significance possiblly.1.3 The result of literature research discovered that the essential study of asthenia cold syndrome had been shifting from the study of sing index to more indexs. But new indexs that were emploied the essential study of asthenia cold syndrome were few, so this means that new ideas was lack. Nowadays, the specificity and sensitivity index of asthenia cold syndrome don't appear. 2 The research of symptomatic rule of asthenia cold syndrome2.1 According to the results of literature research, experts opinion and beforehand investigating, we studied and formulated the clinical epidemiological investigational questionnaire of syndromes of asthenia cold syndrome.The results of clinical epidemiological investigation and analysis of data indicated that this questionnaire had better reliability and validity. And it also had some widespread applicability.2.2 The results of clinical epidemiological investigation signed that the case rate of asthenia cold syndrome in crowd was about 9.946%.2.3 The statistics results shew that the occurrence of asthenia cold syndrome related with such demography characteristic as sex, age, educational background,etc. there had statistical significance(all P<0.01); Among them, fang lao and copiousness was the main risk factor, OR=27.926,which was followed by asthenia cold constitution, reception chill, improper diet, sentiment injury, excessus exertion.2.4 According to the results of cluster analysis and factor analysis of variance, frequent type of syndromes of asthenia cold syndrome were deficiency of kidney-yang(18.828%),deficiency of spleen-yang and kidney-yang(16.318%), deficiency of heart-yang and kidney-yang(14.644%), deficiency of spleen-yang(10.041%), deficiency of heart-yang and spleen-yang(10.041%), deficiency of lung-yang and kidney-yang(8.787%), deficiency of heart-yang(6.276%),deficiency of lung-yang(5.021%), deficiency of liver-yang and spleen-yang(4.184%). analytic result also shew that main two viscus pathopoiesis together is more, occuping 56.904%; sing viscera pathopoiesis occupied 40.167%; Among them, the proportion of deficiency of kidney-yang in asthenia cold syndrome was larger. Deficiency of liver-yang appeared with other viscus together.2.5 Through principal component analysis and logistic stepwise regression analysis, we got cardinal symptoms which had statistics significance in diagnosis of each type of syndrome of asthenia cold syndrome. They were as follows(The main symptoms arranged according to OR value ): Deficiency of spleen-yang: Main symptoms were abdominal distention, loose stool, lassitude, chilly; Secondary symptoms were limbs cold, anorexia, mind weary, aggravation with cold, relief with warm and pressure, pale and enlarged tongue.Deficiency of liver-yang and spleen-yang: Main symptoms were aggravation with cold, anorexia, bilateral hypochondria full, nauseated, eyes blurred; Secondary symptoms were chilly, dizzy, limbs cold, bilateral hypochondria vague pain, hot drink preference, relief with warm and pressure.Deficiency of heart-yang: Main symptoms were hot drink preference, palmus, weakness pulse, zhengzhong; Secondary Symptoms were chilly, limbs cold, short breath, spontaneous perspiration, darkness oral lip, edema of lower limbs, aggravation with cold.Deficiency of lung-yang: Main symptoms were catching a cold readily, cough, whiteness phlegm, aggravation with exhaustoin; Secondary symptoms were chilly, short breath, limbs cold, light body of the tongue.Deficiency of heart-yang and spleen-yang: Main symptoms were deep and delay pulse, palmus, loose stool, abdominal distention, chest depressed;Secondary symptoms were chilly, limbs cold, relief with warm and pressure, head and face puffiness, thinness and whiteness moss.Deficiency of lung-yang and kidney-yang: Main symptoms were acid and weak in waist and knee, cough, whiteness body of the tongue, catching a cold readily, oliguresis; Secondary symptoms were chilly, limbs cold, whiteness phlegm, mind weary, whiteness and greasiness moss.Deficiency of kidney-yang: Main symptoms were urinary frequency, acid and weak in waist and knee, huangbai complexion, whiteness moss; Secondary symptoms were limbs cold, urinary incontinence, multi- nycturia, urine clear, pale and enlarged tongue with indentation, extenuate and weakness pulses or slow and weakness pulses.Deficiency of heart-yang and kidney-yang: Main symptoms were palmus, oliguresis, acid and weak in waist and knee, aggravation with cold, chest pain; Secondary symptoms were chilly, limbs cold, lassitude, edema of lower limbs,droping after urine, extenuate and weakness pulses or deep and weakness pulses.Deficiency of spleen-yang and kidney-yang:Main symptoms were loose stool, huangbai complexion, urinary frequency, acid and weak in waist and knee, gastric cavity vague pain; Secondary symptoms were wangubuhua, limbs cold,lassitude, chilly, hot drink preference,deep and delay puleses.2.6 Synthesizeing the result of statistics and the opinion of experts, we considered chilly, limbs cold as characteristic symptoms of asthenia cold syndrome.3 The metabonomics'research of asthenia cold syndrome3.1 Compared with normal group, there were conspicuous changes in 3.2～3.96ppm, 4.0 ppm, 1.22 ppm, 2.23ppm,2.41 ppm, 2.57 ppmpeak forms of the serum 1HNMR spectra in asthenia cold syndrome group and asthenia pyrosyndrome group .3.2 Through multielement ANOVA: compared with asthenia pyrosyndrome group and normal group, there were obviously low in the contents of serumα-glucose(5.12ppm),β-glucose(3.14ppm,4.56ppm), RegionA(3.2～3.96ppm)(glucose and amino acids), citrate(2.57ppm), succinate(2.23ppm), unsaturated lipid (5.18ppm, 2.63ppm, 2.34ppm, 1.89ppm, 1.62ppm, 1.45 ppm), very low density lipoprotein(1.16ppm), glutamine(2.41ppm), unkown compound(1.79ppm) of asthenia cold syndrome group. there were statistical significance; The contents of serum acetoacetate(2.11ppm), lactate(1.22ppm,4.0ppm) were obviously high, there were statistical significance, too.( P<0.05,P<0.01;P<0.01,P<0.01).In addition,compared with normal group,the contents of valine(0.92ppm), leucine(0.88ppm) and low density lipoprotein(0.75ppm) of asthenia cold syndrome cut down,there were statistical significance (P < 0.05,P < 0.05,P < 0.05),but compared with asthenia pyrosyndrome,there weren't statistical significance(P >0.05,P >0.05,P>0.05).Compared with asthenia cold syndrome and normal group,the integral of oppsing area of serum RegionA and lysine (2.84ppm) of asthenia pyrosyndrome obviously heightened.There were statistical significance(P<0.01,P<0.05; P<0.05,P<0.01).3.3 The changes of genesis hormone of asthenia cold syndrome,asthenia pyrosyndrome and normal groupCompared with asthenia pyrosyndrome group and normal group, there were obviously low in the contents of serum FSH,LH,E2 and T of asthenia cold syndrome,there were statistical significance (P<0.01,P<0.01;P<0.01,P<0.01;P<0.01,P<0.01;P<0.01,P<0.05);The content of serum P of asthenia cold syndrome was lower than asthenia pyrosyndrome group,there was statistical significance(P<0.01),but compared with normal group,there wasn't statistical significance(P>0.05).Compared with asthenia cold syndrome and normal group, the contents of serum LH,T and P of asthenia pyrosyndrome group heightened,there were statistical significance(P < 0.01,P < 0.05;P < 0.01,P < 0.01;P < 0.01,P <0.01);The contents of serum E2 and FSH of asthenia pyrosyndrome group were higher than asthenia cold syndrome,there were statistical significance (P<0.01,P<0.01),but compared with normal group,there weren't statistical significance(P>0.05,P>0.05).3.4 The dependability study of between biomarkers and hormoneAfter analysis of variance,we carried out correlation analysis between all metabolin which had changes in comparision of asthenia pyrosyndrome group, asthenia cold syndrome and normal group and hormone which had changes in them.The results were that there were negative correlation between lactate and FSH and between lactate and LH respectively(r=-0.839,P<0.05;r=-0.865,P<0.05),but there weren't dependablity between lactate and E2 and between lactate and T. There were positive correlation between citrate and FSH and between citrate and LH(r=0.826,P<0.05;r=0.812,P<0.05).There were also positive correlation between succinate and FSH and between succinate and LH(r=0.889,P<0.05;r=0.833,P<0.05),but the two hadn't dependability with E2 and T. Unsaturated lipid,VLDL, acetoacetate, glutamine and 1.79ppm unkown compound hadn't dependability with FSH,LH,E2 and T. conclusion:Through adopted methods of literature's research, clinical epidemiology investigation and metabonomics, we carried out initially systematical research on asthenia cold syndrome. the conclusions were as follows:1 Used the method of literature research, we approached etiological factor and clinical manifestation feature of asthenia cold syndrome, and held its regular pattern of research and developed direction in the whole. This established theoretical and literary foundation for the next study.2 The clinical epidemiology investigation found that case rate of asthenia cold syndrome was about 9.946%; The results of statistical analysis shew that occurrence of asthenia cold syndrome related with such demography feature as sex, age, educational background, vocation, habitation,ect. Fang lao and copiousness was main influential factor of asthenia cold syndrome, which were followed by asthenia cold contitution, reception chill, improper diet, sentiment injury,excessive exertion. On these grounds, we could preclude the occurrence of asthenia cold syndrome directly.3 The frequent type of syndromes in asthenia cold syndrome were deficiency of kidney-yang,deficiency of spleen-yang and kidney-yang, deficiency of heart-yang and kidney-yang, deficiency of spleen-yang, deficiency of heart-yang and spleen-yang, deficiency of lung-yang and kidney-yang, deficiency of heart-yang, deficiency of lung-yang, deficiency of liver-yang and spleen- yang.4 Through literature research, combining opinion of experts and epidemiological investigation,we determined main indexs of differentiation of symptoms and signs of each type of syndrome. Chilly and limbs cold were characteristic symptom of asthenia cold syndrome. This offered information for establishing diagnostic criteria of asthenia cold syndrome.5 Through clinical epidemiology investigation, we obtained methods of investigation, quality control and data processing of large sample clinical epidemiology. We carried out significant exploration on the research of syndromes of traditional Chinese medicine. 6 The dysmetabolism of suger, fat and protein resulted in energy shortage. This was main cause which led to occurrence of asthenia cold syndrome possiblly. We tentatively confirmed that lactate, citrate, succinate, unsaturated lipid, VLDL, acetoacetate, glutamine, 1.79ppm unkown compound could be thought of as biomarkers of asthenia cold syndrome.7 The body of asthenia cold syndrome might be that glycometabolism was blocked, brain's function was damaged.This may influence pituitary and hypothalamus or highest center and make hormone of body low, so shew menstrual disorder.8 Metabonomics analysis was a well developed outlook method which studied syndromes of traditional Chinese medicine. It conduced more overall to understand chemical materials changes of organism, to reveal essence of syndromes of traditional Chinese medicine.