Construction Of OSCC Diagnostic Model Based On Microbiome Metaanalysis And Development Of Oral Probiotic Solid Beverage

The oral cavity is one of the key areas of focus in the Human Microbiome Project.This complex environment harbors diverse microbial communities,which interacts closely with the human health.Oral squamous cell carcinoma(OSCC)is the most common type of oral cancer,and approximately one-third of OSCC patients will develop recurrent OSCC which is life-threatening and incurable.In recent years,the correlation between oral microbiota and OSCC has attracted extensive attention.However,most studies only focus on single-batch samples,which inevitably leads to false positive conclusions.In order to explore the changes in the oral microbiota in OSCC samples and evaluate the reproducibility of these changes across different datasets,this study conducted a meta-analysis of publicly available OSCC datasets.We built a cancer diagnostic model using differential microorganisms identified via our meta-analysis.Additionally,in order to enrich the current product types of oral microbiome preparations,we developed a solid beverage with oral probiotic functions.The main research contents and results are as follows:By using a combination of subject headings and Textwords,literature related to OSCC was screened in Pub Med.Eight publicly available 16 S r RNA datasets were included after evaluating data quality and Metadata,including 268 squamous carcinoma samples from OSCC patients,164 oral samples from healthy sites of OSCC patients,and 107 oral samples from healthy individuals,for a total of 539 samples.The included samples cover almost all sites in the oral cavity,including the gums 、 tongue 、 cheeks etc.The sampling methods include swabs 、 saliva and biopsy,which are commonly used.The samples were sourced from both domestic and foreign regions.QIIME2 was used for preprocessing the included sequence data,and the processed sequence data had high quality and were suitable for subsequent analyses.Using QIIME2 to calculate the Alpha diversity and Beta diversity of microbial samples,and using the lmer function and Adonis function to respectively calculate the overall effect of differences between different samples in each dataset.The results showed that the Alpha diversity of OSCC samples did not show consistent changes in multiple datasets,but the Beta diversity of different sample types showed significant differences.DESeq2 package in R was used to calculate the abundancedifferent OTUs between samples.Using the top 20 differential OTUs with the largest effect sizes to construct a random forest model for distinguishing between squamous carcinoma samples of OSCC patients and oral samples of healthy individuals,with an average AUC of 0.79.Using 46 differential OTUs based on abundance to construct a random forest model for distinguishing between squamous carcinoma samples and healthy site samples from OSCC patients’ oral,with an average AUC of0.72.Both models showed good predictive performance.Microorganisms associated with oral diseases such as dental caries and periodontitis,such as Selenomonas,Campylobacter,Treponema and Catonella were significantly enriched in squamous carcinoma samples.In addition,Fusobacterium and Capnocytophaga,which have been reported to be associated with oral cancer,were significantly enriched in squamous carcinoma samples in at least three datasets.Given the close relationship between oral microbiota and oral health,this study aims to develop a beneficial microbial for oral health.A solid beverage is prepared by compounding xylitol,Fructooligosaccharides,whole milk powder,inulin and Lactobacillus.The formula of the solid beverage is determined by single-factor experiments and orthogonal experiments with sensory evaluation as an indicator,and the formula is as follows: xylitol added at 50%,Fructooligosaccharides added at20%,whole milk powder added at 20%,inulin added at 5%,and probiotic powder added at 5%.The total weight of the finished product is 3 grams.The powder is fine and uniform with white color and has a faint milky fragrance.The brewed solid drink is milky white and has a slightly sweet taste.