Metagenomics Analysis Of Yellow Thick Greasy Fur In Type 2 Diabetic Patients And Healthy People

PurposeObservation is one of the main method of traditional Chinese medicine diagnosis.Tongue diagnosis is an important part of observation.However,at present,the biological basis of tongue diagnosis is still unclear.Clinicians' judgment of tongue coating is often based on personal experience.which is easy to cause ambiguity.Therefore.the standardized research of tongue coating is helpful to promote the scientific and modern diagnosis of traditional Chinese medicine.Diabetes is a chronic metabolic disease with a high incidence.Previous studies have found that yellow-thick-greasy fur is a common tongue manifestation in type 2 diabetic population.This study attempts to analyze the microbial composition of yellow-thick-greasy coatings in type 2 diabetic patients based on metagenomics technology.and to compare with the yellow-thick-greasy coatings of healthy people to explore the biological basis of yellow-thick-greasy coatings in type 2 diabetic patients,and provide scientific basis for clinical diagnosis.MethodsDiabetic and healthy subjects with yellow-thick-greasy fur were recruited from Guang'anmen Hospital of the Chinese Academy of traditional Chinese medicine and Xinjiekou community.There were 15 subjects in total,including 7 patients with type 2 diabetes and 8 healthy people.The tongue coating of subjects were collected 30 minutes after gargling in the morning when they were fasting,and the photos of tongue coating were retained with a portable tongue diagnostic instrument.All the DNA of yellow thick greasy moss of type 2 diabetic patients and healthy people were analyzed by macrogenomic sequencing.The clean data were assembled and predicted by software Megahit and METAProdigal.The representative sequences were assignment for taxonomy against NR database by the DIAMOND software.The taxonomies information included Phylum.Class.Order.Family.Genus,Species.To understand the functions of the differentially expressed genes,the genes were assignment against the ARDB database and KEGG database by using the BLASTP.The genes were also blasted with the CAZy database with the Hmmscan.Results1.There was no significant difference in microbial abundance and species diversity between type 2 diabetes and healthy people.2.Bacterial results:at the phylum level.the abundance of Firmicutes was the highest in the two groups,and the abundance of cyanobacteria in the healthy group was higher than that in the type 2 diabetes group;at the class level,the abundances of actinobacteria and bacteroidia were the highest in the two groups.The abundances of Actinobacteria in the type 2 diabetes group were higher than those in the healthy group;at the order level,bacteroidales,latobacillales neisseriales.Actinomycetales and veillonellales have the highest abundances in the two groups,while chroococcales and frankieles have higher abundances in the healthy group than in the type 2 diabetes group;at the family level.prevotellace,streptocaccae,Neisseriaceae,actinomycetaceae,veillonellaceae and Micrococcaceae had the highest abundance in the two groups.There were 9 families with significant differences between the two groups;at the genus level,Prevotella,Streptoccus,Neisseria,Actinomyces and Veillonella have the highest abundance in the two groups,with a total of 34 genera with significant differences;at the species level,Rothia mucilaginosa,Prevotella melaninogenica,actinomyces sp:ICM47.There are 546 species with significant.3.Results of archaea:Euryarchaeota was the main phylum of Archaea in the two groups,and thaumarchaeota in the healthy group was significantly higher than that in the type 2 diabetes group;4.Results of eukaryote:the main prokaryotes that make up the tongue coating are human cells.In addition,there are some prokaryotes that parasitiz'e the human body;5.Results of virus:human endogenous retrovirus is the main virus in tongue coating;6.Type 2 diabetes mellitus patients have higher levels of antibiotic resistance gene expression,carbohydrate enzyme gene expression and functional clustering gene expression than healthy people.ConclusionsThere is no significant difference in the abundance and species diversity of yellow-thick-greasy fur microorganisms between type 2 diabetes and healthy people.but there are different species at different levels,among which Actinobacteria,leptotrichiaceae,Leptotrichia may be the possible bacterial biomarkers of type 2 diabetes.The expression of higher level antibiotic resistance genes,carbohydrate-active enzymes genes and functional clustering genes may be the characteristics of type 2 diabetes patients in distinguishing healthy people.