Fecal-associated microbiome differences between traditional Chinese medicine qi-deficiency and balanced constitutions

Objective:To explore the structural and functional characteristics of the fecal-associated microbiome(FAM)in a traditional Chinese medicine(TCM)qi-deficiency constitution(QDC)by comparing with balanced constitution(BC)and screen the related biomarkers.Methods:In this cross-sectional study,the TCM constitutions of subjects were determined based on published the Classification and Determination of constitution in TCM and further confirmed by a TCM clinician.Clinical characteristics were recorded,and fecal samples were collected for 16S rDNA sequencing using the Illumina Miseq platform.The FAM structure was described using alpha-diversity indexes,beta-diversity indexes,and the relative abundances of the dominant taxa.Differences in the FAM distribution and function were analyzed with a Wilcoxon rank-sum test,MetagenomeSeq,and LEfSe analysis,after which a receiver operating characteristic curve based on the specific operational taxonomic units(OTUs)was constructed to calculate the area under the curve.Results:Our study population was composed of 22 BCs and 9 QDCs.There were no significant differences between the two groups in the distribution of clinical characteristics or alpha-diversity indexes,except for the sweets preference and blood glucose level.In principal coordinate analysis and partial least squares discriminant analysis,the bacterial communities in the BC group samples and QDC group samples clustered separately.Notably,there were 214 OTUs significantly distributed between groups in the MetagenomeSeq analysis,200 OTUs identified by the Wilcoxon rank-sum test,and 6 OTUs found by the LEfSe analysis.Predicted function analysis revealed that six metabolic pathways were distinctly distributed between the two groups.The area under the curve for the receiver operating characteristic curve based on the four specific OTUs was 0.88.Conclusion:Unique FAM structural and related functional characteristics are displayed in individuals with a QDC,and four specific OTUs could be used as QDC biomarkers to assist in clinical diagnosis.

Fecal-associated microbiome differences between phlegm-dampness constitution and balanced constitution

Objective: This study aimed to explore the structural and functional characteristics of the fecalassociated microbiome(FAM) between the phlegm-dampness constitution(PDC) and balanced constitution(BC), and to screen the related specific operational taxonomic unit(OTU) biomarkers.Methods: This was a cross-sectional study. After strictly identifying the constitution of subjects, their clinical index was recorded and counted. Fecal samples were collected for 16S rDNA sequencing. Alpha diversity, beta diversity, and the relative abundance of dominant bacterial taxa were used to describe the FAM structure, and the Wilcoxon rank-sum test, Metagenome Seq, and linear discriminant analysis effect size(LEf Se) were used to screen specific bacterial taxa. Specific OTUs were screened to construct receiver operating characteristic(ROC) curves.Results: Thirty-two subjects were enrolled, including 22 subjects with BC and 10 subjects with PDC.There were significant differences in cold preference, levels of aspartate transaminase, β2-microglobulin,and creatine kinase MB, and alpha diversity indices(Shannon and Shannoneven) between the two groups. In principal coordinate analysis by abund-jaccard distance measure and partial least squares discriminant analysis, bacterial communities clustered separately between the two groups. Furthermore,based on Metagenome Seq, LEf Se, and the Wilcoxon rank-sum test, a total of 43, 18, and 130 OTUs were differentially distributed between BC group and PDC group, respectively, and OTU200, OTU133, and OTU353 were screened when P≤.01. The area under the ROC curve constructed from the 3 selected OTUs was 0.93.Conclusion: The FAM structure and related functional characteristics of the PDC group differed from those of the BC group. In particular, OTU200, OTU133, and OTU353 can be used as unique markers of PDC to assist clinical diagnosis.

Discrimination of TCM constitutions by biochemical and routine urine indexes

Objective:To investigate whether the specific traditional Chinese medicine(TCM)constitution of individuals can be defined by certain biological indexes instead of answering the questionnaire,and to explore the possibility of discriminating nine TCM constitutions from each other simultaneously using biological indexes.Methods:Blood and urine samples from 152 individuals with nine TCM constitutions were collected,and the related biological indexes were analyzed combining ANOVA,multiple comparison,discriminant analysis,and support vector machine.Results:We found that 4 out of 24 blood routine indexes,7 out of 10 urine routine indexes,and 12 out of 32 biochemical indexes showed differences among the constitutions.High-sensitivity C-reactive protein,apolipoprotein A1,and alkaline phosphatase were potential candidates for screening out individuals with unbalanced constitutions.Combining uric acid,high-density lipoprotein,apolipoprotein A1,creatine kinase,total protein,aspartate aminotransferase,total bile acid,dehydrogenase,sodium,and calcium levels had the potential to directly distinguish the nine TCM constitutions from each other.Among these indexes,the highest ratio of discriminant analysis between two constitutions was 95.5%,while the lowest was 66.1%.Conclusion:Our results suggest that some biochemical and urine indexes are related to various TCM constitutions,and thus they have the potential to be used for TCM constitution classification.

Structure and function of the fecal-associated microbiome in qi stagnation constitution

Objective:To clarify the structural and functional characteristics of the gut microbiota in individuals with qi stagnation constitution(QSC)and identify the potential biomarkers related to QSC.Methods:This cross-sectional study involved individuals with QSC and balanced constitution(BC)confirmed by TCM clinicians.The clinical features were recorded,and fecal samples were collected for 16S rDNA sequencing.The structure of the fecal-associated microbiome(FAM)was described by the alpha-diversity indexes,beta-distances,and relative abundances of dominant taxa.The differences in FAM distribution were analyzed by the Wilcoxon rank-sum test,MetagenomeSeq,and LEfSe analysis.The 16S rDNA gene sequences were assigned to the KEGG dataset to predict the functional information of bacterial metabolic pathways by using PICRUSt.Differences in functional pathways between groups were assessed with the Wilcoxon rank-sum test.The ROC curve based on specific operational taxonomic units(OTUs)was constructed,and the AUC was calculated.Results:Twenty-two individuals with BC and 8 with QSC were recruited.Significant differences between the two groups were found in body mass index,health status,and low-density lipoprotein,etc.There was no significant difference in the alpha-diversity index.PCoA showed no evident clustering of bacterial communities according to constitutions.Bacteroidaceae,Lachnospiraceae,Ruminococcaceae,and Prevotellaceae were the four common bacteria with high abundances.Notably,MetagenomeSeq,LEfSe analysis,and the Wilcoxon rank-sum test identified significantly different distributions of 66,42,and 36 OTUs,respectively.Predictive function analysis showed that 13 metabolic pathways were significantly differentially distributed,including those related to fatty acid synthesis.Five specific OTUs were selected as potential biomarkers of a QSC,and the AUC was 0.94.Conclusion:Individuals with QSC have unique FAM structure and related functional characteristics.Five specific OTUs were identified to serve as potentially effective biomarkers related to QSC.