A Study On The Macroscopic And Microscopic Of Dampness Syndrome In Chinese Medicine

ObjectiveSyndrome differentiation and treatment are the characteristics of Chinese medicine(CM).Syndrome differentiation therapy is based on "syndrome".Syndrome is the key scientific problem of the theoretical system of CM,which is one of the core contents connecting the basic theory and clinical practice of CM.The modern scientific connotation of CM syndromes can be revealed from different perspectives,including the combination of macroscopic and microscopic features,which can be applied.Dampness syndrome is one of the most common CM syndromes.It has been gradually considered as the core pathogenesis of many chronic diseases.Presently,the researches on dampness syndrome in Chinese medicine focus on diagnosis,clinical characteristics,distinguishing features of different diseases,and the characteristics of disease-specific population from microscopic intestinal flora and laboratory indicators.Few studies combine macro performance with microscopic laboratory indicators to simultaneously clarify the biological characteristics and peculiarity of the CM dampness syndrome.We aim to explore the clinical characteristics and micro indicators of the CM dampness syndrome,taking a disease as an example to explore,analyze and reveal the features of the CM dampness syndrome.MethodsPart Ⅰ:A macroscopic study on the CM dampness syndromeA total of 678 patients with eczema/atopic dermatitis(AD)with a preestimated sample size were to be enrolled nationwide based on a multicenter disease registry cohort with CM dampness syndrome as the study factor.We used the clinical epidemiological investigation method to describe the distribution characteristics of CM symptoms and symptoms of the patients.Univariate analysis was performed on the macroscopic syndromes variables to analyze the correlation between CM dampness syndrome and these variables.According to disease characteristics and clinical practice,variables with P≤0.1 in univariate analysis were included in multivariate Logistic regression model as covariates to analyze important factors affecting the differentiation of CM dampness syndrome and non-dampness syndrome,and to explore the association between CM dampness syndrome and state of disease,clinical features,life behavior and quality of life.For patients with CM non-dampness syndrome,two-step clustering method was used to classify different types of CM syndromes,so as to summarize and describe the types of syndromes and their macroscopic symptoms.Part Ⅱ:A microscopic study of the CM dampness syndromeSkin microbiotic samples from the CM dampness syndrome and non-dampness syndrome patients were collected from the registry cohort,and skin microbiotic samples from healthy controls were collected simultaneously.Sequencing of the 16S rDNA of skin bacteria were conducted to find the skin microbial composition in different CM syndromes.Different analysis tools,statistical methods and machine learning methods including random forest were used to perform species clustering,diversity analysis,complexity analysis and sample classification of skin microorganisms between CM dampness syndrome and non-dampness syndrome.ResultsPart Ⅰ:A macroscopic study on the CM dampness syndrome1.A total of 679 patients with eczema/AD were enrolled from 14 hospitals in China,including 218 patients with CM dampness syndrome and 461 patients with CM non-dampness syndrome.The average age of the patients was 44.14,and most of them were male(395 cases,58.2%).The education level of the patients was mainly undergraduate(373 cases,54.9%).The mean Eczema area and severity index(EASI)total score was 3.57[standard deviation(SD)=4.09].The mean body surface area(BSA)was 5.15(SD=7.57),and the mean investigator global score was 2.53(SD=0.64).The mean pruritus numerical score was 5.24(SD=2.25).The scores of Skindex were 37.47(SD=23.25)in emotional domain,40.47(SD=18.36)in symptom domain,27.27(SD=23.24)in functioning domain and 33.97(SD=20.53)in overall.The EQ-5D utility index values of different dimensions were:mobility dimension average 0.004,self-care dimension average 0.003,daily activities dimension average 0.005,pain or discomfort dimension average 0.06,anxiety or depression dimension average 0.043,utility index set average 0.885.2.The results of univariate analysis found that,Statistical difference of marital status,occupation,smoking habits of family members,acid-base of skin cleansing products,vitamin D intake,weight at birth,mother’s age at birth,feeding regimens,breast-feed duration,heterologous protein allergies,disease course(month),erythema(head,upper limbs and trunk),infiltration or papule(head parts,upper limbs,and trunk),exfdiation(head,upper limbs,and trunk),lichenification(trunk),EASI score(head,neck,upper limbs and trunk),EASI total score,IGA score,BSA,Skindex-29 emotion score and Skindex-29 overall score between the patients with CM dampness syndrome and non-dampness syndrome(P<0.05).There was no statistically significant difference in age,BMI,education level,history of smoking,drinking,personal hygiene habits,exercise routines,maternal history,family structure,pets,adolescent obesity,acetaminophen medication history,parental allergy,food allergy,drug allergy,history of contact allergy,history of asthma or allergic rhinitis,course of disease,family history of eczema/AD and comorbidity(P>0.05).3.Multivariate Logistic regression analysis found that smoking habits of family members(P=0.013),acid-base of skin cleansing products(P=0.001),weight at birth(P=0.017),breastfeeding duration(P=0.012),BSA(P=0.013)and Skindex-29 emotion score(P=0.008)were independent related factors of CM dampness syndrome.4.Frequency analysis found that the most common CM symptoms in dampness syndrome patients are fatigue,loose stool or diarrhea or postprandial diarrhea,body drowsiness,insomnia and dreamful sleep,inappetence,restlessness and irritability,shortness of breath and lazy word.And they have white moss,greasy tongue fur,tongue with tooth prints,light tongue,and soggy pulse.In addition to the features of dampness,patients with CM dampness syndrome are also with deficiency symptom.5.The two-step clustering method was used to classify CM non-dampness syndrome,which can be divided into spleen deficiency and qi depression and yin deficiency and qi depression.Part Ⅱ:A microscopic study of the CM dampness syndrome1.We had completed two rounds of pre-experiments for skin microbial sample collection.The sampling process was revised and the standard operating procedure of skin microbiota in patients with eczema/AD and healthy subjects was determined.2.In this study,skin microbiota samples were collected from 19 patients with CM dampness syndrome and 20 patients with non-dampness syndrome in the eczema/AD cohort,and 21 age-and sex-matched healthy controls were recruited.3.The diversity of the skin microbial community in CM dampness syndrome group and non-dampness syndrome group were found higher than health controls.There was a difference in species richness between the skin microbiota of the CM dampness syndrome group and the non-dampness syndrome group,but there was no difference in the relative abundance.4.Results of skin microbiological analysis in CM dampness syndrome and non-dampness syndrome were as follows.The Staphylococcus species in the CM dampness syndrome group and non-dampness syndrome group were significantly higher than those in the healthy control.Pseudomonas and Staphylococcus were the dominant genera in the CM dampness syndrome group and non-dampness syndrome group.At genus level,the abundance of Ralstonia,Acinetobacter and Vibrio in the skin microorganisms of patients with CM dampness syndrome group was higher than that of non-dampness syndrome group.The abundance of Microbacterium,Methyloversatilis,Paracoccus and Deinococcus of the CM non-dampness syndrome group was higher than that of dampness syndrome group.ROC analysis showed that random forest performed by selecting 5-200 important bacterial genera could achieve good predictive diagnostic value for distinguishing CM dampness syndrome and non-dampness syndrome(AUC:90.58-93.95%).When the 10 most important variables were selected to build a random forest classifier(AUC=92.96%),the genus in the model was Vibrio,Paracoccus,Microbacterium,Methyloversatilis,Kytococcus,Gordonia,Tsukamurella,Exiguobacterium,Dermacoccus,Acinetobacter.Acinetobacter,Paracoccus and Methyloversatilis were found to be coincident between the results of random forest model and those screened by LEfSe analysis.5.Results of skin microbiological analysis in CM dampness syndrome and non-dampness syndrome(different parts of body skin)were as follows.There is no difference in alpha diversity between different parts of the body skin of patients with different CM syndrome.At genus level,the abundance of Ralstonia,Acinetobacter,and Vibrio in the skin microorganisms of patients with CM dampness syndrome group was higher than that of non-dampness syndrome group.The abundance of Kocuria,Methyloversatilis and Paracoccus of the CM non-dampness syndrome group was higher than that of dampness syndrome group.ROC analysis showed that random forest performed by selecting 20-200 important bacterial genera could achieve good predictive diagnostic value for distinguishing CM dampness syndrome and non-dampness syndrome(AUC:95.09-96.03%).When the 20 most important variables were selected to build a random forest classifier(AUC=95.09%),the genus in the model was Vibrio,Tsukamurella,Paracoccus,Microbacterium,Methyloversatilis,Kytococcus,Gordonia,Acinetobacter,Lactobacillus,Massilia,Methylobacterium_Methylorubrum,Pseudoalteromonas,Exiguobacterium,Uruburuella,Roseomonas,Aeromonas,Phreatobacter,Finegoldia,Bacillus and A4b.The results of the random forest model and the genus screened by LEfSe analysis overlap with Acinetobacter,Paracoccus and Methyloversatilis.6.The differential bacteria between CM dampness syndrome group(lesion parts)and non-dampness syndrome group(lesion parts)are Acinetobacter,Paracoccus and Methyloversatilis,which is the same as the differential bacteria between the CM dampness syndrome group(lesion and non-lesion)and the non-dampness syndrome group(lesion and non-lesion).ConclusionIn this cross-sectional study of patients with eczema/AD,smoking habits of family members,acid-base of skin cleansing products,weight at birth,breastfeeding duration,BSA and Skindex-29 emotion score were independent related factors for dampness syndrome differentiation at the macroscopic level.There was a positive correlation between family members smoking and CM dampness syndrome.Acid-base of skin cleansing products,birth weight and BSA were negatively correlated with CM dampness syndrome.Breastfeeding for 6-12 months has the strongest correlation with the CM dampness syndrome.With the increase of breastfeeding time,the correlation between breastfeeding and dampness syndrome gradually decreased.Emotional impairment was positively correlated with the CM dampness syndrome.The CM symptoms of patients with dampness syndrome were mainly a series of manifestations of dampness.It should be also noted that many patients with dampness syndrome were accompanied by deficiency syndrome.At the microscopic perspective,Acinetobacter,Paracoccus and Methylotrophum were the differential bacteria in skin microbiota between the CM dampness syndrome group and the non-dampness syndrome group,no matter in the skin lesion groups or which groups included lesion and non-lesion parts.It embodies the features of CM theory that holistic syndrome differentiation as the basis,local syndrome differentiation is part of holistic syndrome differentiation,and both of them are inseparable.The experience,method and model of this study can be used as a reference for macro and micro CM syndrome differentiation research in the future.