Distribution characteristics of oral microbiota and its relationship with intestinal microbiota in patients with type 2 diabetes mellitus.

Introduction: Type 2 diabetes mellitus (T2DM) has a high incidence rate globally, increasing the burden of death, disability, and the economy worldwide. Previous studies have found that the compositions of oral and intestinal microbiota changed respectively in T2DM; whether the changes were associated or interacted between the two sites and whether there were some associations between T2DM and the ectopic colonization of oral microbiota in the gut still need to be identified. Research design and methods: We performed a cross-sectional observational study; 183 diabetes and 74 controls were enrolled. We used high-throughput sequencing technology to detect the V3-V4 region of 16S rRNA in oral and stool samples. The Source Tracker method was used to identify the proportion of the intestinal microbiota that ectopic colonized from the oral cavity. Results: The oral marker bacteria of T2DM were found, such as Actinobacteria, Streptococcus, Rothia, and the intestinal marker bacteria were Bifidobacterium, Streptococcus, and Blautia at the genus level. Among them, Actinobacteria and Blautia played a vital role in different symbiotic relationships of oral and intestinal microbiota. The commonly distributed bacteria, such as Firmicutes, Bacteroidetes, and Actinobacteria, were found in both oral and intestine. Moreover, the relative abundance and composition of bacteria were different between the two sites. The glycine betaine degradation I pathway was the significantly up-regulated pathway in the oral and intestinal flora of T2DM. The main serum indexes related to oral and intestinal flora were inflammatory. The relative abundance of Proteobacteria in the intestine and the Spirochete in oral was positively correlated, and the correlation coefficient was the highest, was 0.240 (P<0.01). The proportion of ectopic colonization of oral flora in the gut of T2DM was 2.36%. Conclusion: The dysbacteriosis exited in the oral and intestine simultaneously, and there were differences and connections in the flora composition at the two sites in T2DM. Ectopic colonization of oral flora in the intestine might relate to T2DM. Further, clarifying the oral-gut-transmitting bacteria can provide an essential reference for diagnosing and treating T2DM in the future.

Deep Learning Multi-label Tongue Image Analysis and Its Application in a Population Undergoing Routine Medical Checkup.

Background: Research on intelligent tongue diagnosis is a main direction in the modernization of tongue diagnosis technology. Identification of tongue shape and texture features is a difficult task for tongue diagnosis in traditional Chinese medicine (TCM). This study aimed to explore the application of deep learning techniques in tongue image analyses. Methods: A total of 8676 tongue images were annotated by clinical experts, into seven categories, including the fissured tongue, tooth-marked tongue, stasis tongue, spotted tongue, greasy coating, peeled coating, and rotten coating. Based on the labeled tongue images, the deep learning model faster region-based convolutional neural networks (Faster R-CNN) was utilized to classify tongue images. Four performance indices, i.e., accuracy, recall, precision, and F1-score, were selected to evaluate the model. Also, we applied it to analyze tongue image features of 3601 medical checkup participants in order to explore gender and age factors and the correlations among tongue features in diseases through complex networks. Results: The average accuracy, recall, precision, and F1-score of our model achieved 90.67%, 91.25%, 99.28%, and 95.00%, respectively. Over the tongue images from the medical checkup population, the model Faster R-CNN detected 41.49% fissured tongue images, 37.16% tooth-marked tongue images, 29.66% greasy coating images, 18.66% spotted tongue images, 9.97% stasis tongue images, 3.97% peeled coating images, and 1.22% rotten coating images. There were significant differences in the incidence of the fissured tongue, tooth-marked tongue, spotted tongue, and greasy coating among age and gender. Complex networks revealed that fissured tongue and tooth-marked were closely related to hypertension, dyslipidemia, overweight and nonalcoholic fatty liver disease (NAFLD), and a greasy coating tongue was associated with hypertension and overweight. Conclusion: The model Faster R-CNN shows good performance in the tongue image classification. And we have preliminarily revealed the relationship between tongue features and gender, age, and metabolic diseases in a medical checkup population.

A multi-step approach for tongue image classification in patients with diabetes.

BACKGROUND: In China, diabetes is a common, high-incidence chronic disease. Diabetes has become a severe public health problem. However, the current diagnosis and treatment methods are difficult to control the progress of diabetes. Traditional Chinese Medicine (TCM) has become an option for the treatment of diabetes due to its low cost, good curative effect, and good accessibility. OBJECTIVE: Based on the tongue images data to realize the fine classification of the diabetic population, provide a diagnostic basis for the formulation of individualized treatment plans for diabetes, ensure the accuracy and consistency of the TCM diagnosis, and promote the objective and standardized development of TCM diagnosis. METHODS: We use the TFDA-1 tongue examination instrument to collect the tongue images of the subjects. Tongue Diagnosis Analysis System (TDAS) is used to extract the TDAS features of the tongue images. Vector Quantized Variational Autoencoder (VQ-VAE) extracts VQ-VAE features from tongue images. Based on VQ-VAE features, K-means clustering tongue images. TDAS features are used to describe the differences between clusters. Vision Transformer (ViT) combined with Grad-weighted Class Activation Mapping (Grad-CAM) is used to verify the clustering results and calculate positioning diagnostic information. RESULTS: Based on VQ-VAE features, K-means divides the diabetic population into 4 clusters with clear boundaries. The silhouette, calinski harabasz, and davies bouldin scores are 0.391, 673.256, and 0.809, respectively. Cluster 1 had the highest Tongue Body L (TB-L) and Tongue Coating L (TC-L) and the lowest Tongue Coating Angular second moment (TC-ASM), with a pale red tongue and white coating. Cluster 2 had the highest TC-b with a yellow tongue coating. Cluster 3 had the highest TB-a with a red tongue. Group 4 had the lowest TB-L, TC-L, and TB-b and the highest Per-all with a purple tongue and the largest tongue coating area. ViT verifies the clustering results of K-means, the highest Top-1 Classification Accuracy (CA) is 87.8%, and the average CA is 84.4%. CONCLUSIONS: The study organically combined unsupervised learning, self-supervised learning, and supervised learning and designed a complete diabetic tongue image classification method. This method does not rely on human intervention, makes decisions based entirely on tongue image data, and achieves state-of-the-art results. Our research will help TCM deeply participate in the individualized treatment of diabetes and provide new ideas for promoting the standardization of TCM diagnosis.

Relationships Between Diurnal Changes of Tongue Coating Microbiota and Intestinal Microbiota.

The oral cavity and the intestine are the main distribution locations of human digestive bacteria. Exploring the relationships between the tongue coating and gut microbiota, the influence of the diurnal variations of the tongue coating microbiota on the intestinal microbiota can provide a reference for the development of the disease diagnosis and monitoring, as well as the medication time. In this work, a total of 39 healthy college students were recruited. We collected their tongue coating microbiota which was collected before and after sleep and fecal microbiota. The diurnal variations of tongue coating microbiota are mainly manifested on the changes in diversity and relative abundance. There are commensal bacteria in the tongue coating and intestines, especially Prevotella which has the higher proportion in both sites. The relative abundance of Prevotella in the tongue coating before sleep has a positive correlation with intestinal Prevotella; the r is 0.322 (p < 0.05). Bacteroides in the intestine had the most bacteria associated with the tongue coating and had the highest correlation coefficient with Veillonella in the oral cavity, which was 0.468 (p < 0.01). These results suggest that the abundance of the same flora in the two sites may have a common change trend. The SourceTracker results show that the proportion of intestinal bacteria sourced from tongue coating is less than 1%. It indicates that oral flora is difficult to colonize in the intestine in healthy people. This will provide a reference for the study on the oral and intestinal microbiota in diseases.

Correlation between Serum Magnesium Level and Cardiac Valve Calcification in Patients with Chronic Kidney Disease.

BACKGROUND: The aim of the study was to investigate the correlation between serum magnesium (Mg) level and cardiac valve calcification (CVC) in patients with chronic kidney disease (CKD). METHODS: A total of 232 CKD patients hospitalized from August 2016 to December 2020 were divided into CVC and non-CVC groups. Their clinical data and laboratory examination results were compared, the risk factors for CVC in CKD patients were explored using logistic regression analysis, and Spearman's method was used to analyze the correlation between serum Mg level and CVC degree. According to the tertiles of mean serum Mg level, they were assigned into low serum Mg group (≤ 0.96 mmol/L), middle serum Mg group (0.97 - 1.07 mmol/L), and high serum Mg group (≥ 1.08 mmol/L). The relationship of serum Mg level with CVC risk in CKD patients was analyzed through the Cox regression model, and a prediction model was established using independent risk factors. RESULTS: Long CKD duration, low serum Mg level, high serum phosphorus (P) level, and high CKD stage were independent risk factors for CVC. Serum Mg level was significantly negatively correlated with the severity of CVC (r = -0.743, p < 0.05). The risk of CVC was significantly higher in low serum Mg group than that in high serum Mg group [hazard ratio (HR) = 2.852, 95% confidence interval (CI): 1.325 - 6.432, p = 0.005]. A CVC prediction model was established based on independent risk factors as follows: CVC predictive value = EXP [0.491 - 0.546 (CKD duration) - 0.454 (serum P level) + 2.145 (serum Mg level) - 0.812 (CKD stage)]/1 + EXP [0.491 - 0.546 (CKD duration) - 0.454 (serum P level) + 2.145 (serum Mg level) - 0.812 (CKD stage)]. The area under curve of the model was 0.750 (95% CI: 0.822 - 0.965), and that of the CVC prediction model for CKD patients in test set was 0.774 (95% CI: 0.761 - 0.975), showing no significant difference from that in training set (p > 0.05). CONCLUSIONS: Low serum Mg level serves as an independent risk factor for CVC in CKD patients, and may increase the risk of CVC. Therefore, the serum Mg level in CKD patients should be corrected timely in clinical practice.

Application of computer tongue image analysis technology in the diagnosis of NAFLD.

Nonalcoholic fatty liver disease (NAFLD), a leading cause of chronic hepatic disease, can progress to liver fibrosis, cirrhosis, and hepatocellular carcinoma. Therefore, it is extremely important to explore early diagnosis and screening methods. In this study, we developed models based on computer tongue image analysis technology to observe the tongue characteristics of 1778 participants (831 cases of NAFLD and 947 cases of non-NAFLD). Combining quantitative tongue image features, basic information, and serological indexes, including the hepatic steatosis index (HSI) and fatty liver index (FLI), we utilized machine learning methods, including Logistic Regression, Support Vector Machine (SVM), Random Forest (RF), Gradient Boosting Decision Tree (GBDT), Adaptive Boosting Algorithm (AdaBoost), Naïve Bayes, and Neural Network for NAFLD diagnosis. The best fusion model for diagnosing NAFLD by Logistic Regression, which contained the tongue image parameters, waist circumference, BMI, GGT, TG, and ALT/AST, achieved an AUC of 0.897 (95% CI, 0.882-0.911), an accuracy of 81.70% with a sensitivity of 77.62% and a specificity of 85.22%; in addition, the positive likelihood ratio and negative likelihood ratio were 5.25 and 0.26, respectively. The application of computer intelligent tongue diagnosis technology can improve the accuracy of NAFLD diagnosis and may provide a convenient technical reference for the establishment of early screening methods for NAFLD, which is worth further research and verification.

Tongue image quality assessment based on a deep convolutional neural network.

BACKGROUND: Tongue diagnosis is an important research field of TCM diagnostic technology modernization. The quality of tongue images is the basis for constructing a standard dataset in the field of tongue diagnosis. To establish a standard tongue image database in the TCM industry, we need to evaluate the quality of a massive number of tongue images and add qualified images to the database. Therefore, an automatic, efficient and accurate quality control model is of significance to the development of intelligent tongue diagnosis technology for TCM. METHODS: Machine learning methods, including Support Vector Machine (SVM), Random Forest (RF), Gradient Boosting Decision Tree (GBDT), Adaptive Boosting Algorithm (Adaboost), Naïve Bayes, Decision Tree (DT), Residual Neural Network (ResNet), Convolution Neural Network developed by Visual Geometry Group at University of Oxford (VGG), and Densely Connected Convolutional Networks (DenseNet), were utilized to identify good-quality and poor-quality tongue images. Their performances were made comparisons by using metrics such as accuracy, precision, recall, and F1-Score. RESULTS: The experimental results showed that the accuracy of the three deep learning models was more than 96%, and the accuracy of ResNet-152 and DenseNet-169 was more than 98%. The model ResNet-152 obtained accuracy of 99.04%, precision of 99.05%, recall of 99.04%, and F1-score of 99.05%. The performances were better than performances of other eight models. The eight models are VGG-16, DenseNet-169, SVM, RF, GBDT, Adaboost, Naïve Bayes, and DT. ResNet-152 was selected as quality-screening model for tongue IQA. CONCLUSIONS: Our research findings demonstrate various CNN models in the decision-making process for the selection of tongue image quality assessment and indicate that applying deep learning methods, specifically deep CNNs, to evaluate poor-quality tongue images is feasible.

Establishment of noninvasive diabetes risk prediction model based on tongue features and machine learning techniques.

BACKGROUND: Diabetes is a chronic noncommunicable disease with high incidence rate. Diabetics without early diagnosis or standard treatment may contribute to serious multisystem complications, which can be life threatening. Timely detection and intervention of prediabetes is very important to prevent diabetes, because it is inevitable in the development and progress of the disease. OBJECTIVE: Our objective was to establish the predictive model that can be applied to evaluate people with blood glucose in high and critical state. METHODS: We established the diabetes risk prediction model formed by a combined TCM tongue diagnosis with machine learning techniques. 1512 subjects were recruited from the hospital. After data preprocessing, we got the dataset 1 and dataset 2. Dataset 1 was used to train classical machine learning model, while dataset 2 was used to train deep learning model. To evaluate the performance of the prediction model, we used Classification Accuracy(CA), Precision, Recall, F1-score, Precision-Recall curve(P-R curve), Area Under the Precision-Recall curve(AUPRC), Receiver Operating Characteristic curve(ROC curve), Area Under the Receiver Operating Characteristic curve(AUROC), then selected the best diabetes risk prediction model. RESULTS: On the test set of dataset 1, the CA of non-invasive Stacking model was 71 %, micro average AUROC was 0.87, macro average AUROC was 0.84, and micro average AUPRC was 0.77. In the critical blood glucose group, the AUROC was 0.84, AUPRC was 0.67. In the high blood glucose group, AUROC was 0.87, AUPRC was 0.83. On the validation set of dataset 2, the CA of ResNet50 model was 69 %, micro average AUROC was 0.84, macro average AUROC was 0.83, and micro average AUPRC was 0.73. In the critical blood glucose group, AUROC was 0.88, AUPRC was 0.71. In the high blood glucose group, AUROC was 0.80, AUPRC was 0.76. On the test set of dataset 2, the CA of ResNet50 model was 65 %, micro average AUROC was 0.83, macro average AUROC was 0.82, and micro average AUPRC was 0.71. In the critical blood glucose group, the prediction of AUROC was 0.84, AUPRC was 0.60. In the high blood glucose group, AUROC was 0.87, AUPRC was 0.71. CONCLUSIONS: Tongue features can improve the prediction accuracy of the diabetes risk prediction model formed by classical machine learning model significantly. In addition to the excellent performance, Stacking model and ResNet50 model which were recommended had non-invasive operation and were easy to use. Stacking model and ResNet50 model had high precision, low false positive rate and low misdiagnosis rate on detecting hyperglycemia. While on detecting blood glucose value in critical state, Stacking model and ResNet50 model had a high sensitivity, a low false negative rate and a low missed diagnosis rate. The study had proved that the differential changes of tongue features reflected the abnormal glucose metabolism, thus the diabetes risk prediction model formed by a combined TCM tongue diagnosis and machine learning technique was feasible.

A tongue features fusion approach to predicting prediabetes and diabetes with machine learning.

BACKGROUND: Diabetics has become a serious public health burden in China. Multiple complications appear with the progression of diabetics pose a serious threat to the quality of human life and health. We can prevent the progression of prediabetics to diabetics and delay the progression to diabetics by early identification of diabetics and prediabetics and timely intervention, which have positive significance for improving public health. OBJECTIVE: Using machine learning techniques, we establish the noninvasive diabetics risk prediction model based on tongue features fusion and predict the risk of prediabetics and diabetics. METHODS: Applying the type TFDA-1 Tongue Diagnosis Instrument, we collect tongue images, extract tongue features including color and texture features using TDAS, and extract the advanced tongue features with ResNet-50, achieve the fusion of the two features with GA_XGBT, finally establish the noninvasive diabetics risk prediction model and evaluate the performance of testing effectiveness. RESULTS: Cross-validation suggests the best performance of GA_XGBT model with fusion features, whose average CA is 0.821, the average AUROC is 0.924, the average AUPRC is 0.856, the average Precision is 0.834, the average Recall is 0.822, the average F1-score is 0.813. Test set suggests the best testing performance of GA_XGBT model, whose average CA is 0.81, the average AUROC is 0.918, the average AUPRC is 0.839, the average Precision is 0.821, the average Recall is 0.81, the average F1-score is 0.796. When we test prediabetics with GA_XGBT model, we find that the AUROC is 0.914, the Precision is 0.69, the Recall is 0.952, the F1-score is 0.8. When we test diabetics with GA_XGBT model, we find that the AUROC is 0.984, the Precision is 0.929, the Recall is 0.951, the F1-score is 0.94. CONCLUSIONS: Based on tongue features, the study uses classical machine learning algorithm and deep learning algorithm to maximum the respective advantages. We combine the prior knowledge and potential features together, establish the noninvasive diabetics risk prediction model with features fusion algorithm, and detect prediabetics and diabetics noninvasively. Our study presents a feasible method for establishing the association between diabetics and the tongue image information and prove that tongue image information is a potential marker which facilitates effective early diagnosis of prediabetics and diabetics.

Clinical data mining on network of symptom and index and correlation of tongue-pulse data in fatigue population.

BACKGROUND: Fatigue is a kind of non-specific symptom, which occurs widely in sub-health and various diseases. It is closely related to people's physical and mental health. Due to the lack of objective diagnostic criteria, it is often neglected in clinical diagnosis, especially in the early stage of disease. Many clinical practices and researches have shown that tongue and pulse conditions reflect the body's overall state. Establishing an objective evaluation method for diagnosing disease fatigue and non-disease fatigue by combining clinical symptom, index, and tongue and pulse data is of great significance for clinical treatment timely and effectively. METHODS: In this study, 2632 physical examination population were divided into healthy controls, sub-health fatigue group, and disease fatigue group. Complex network technology was used to screen out core symptoms and Western medicine indexes of sub-health fatigue and disease fatigue population. Pajek software was used to construct core symptom/index network and core symptom-index combined network. Simultaneously, canonical correlation analysis was used to analyze the objective tongue and pulse data between the two groups of fatigue population and analyze the distribution of tongue and pulse data. RESULTS: Some similarities were found in the core symptoms of sub-health fatigue and disease fatigue population, but with different node importance. The node-importance difference indicated that the diagnostic contribution rate of the same symptom to the two groups was different. The canonical correlation coefficient of tongue and pulse data in the disease fatigue group was 0.42 (P < 0.05), on the contrast, correlation analysis of tongue and pulse in the sub-health fatigue group showed no statistical significance. CONCLUSIONS: The complex network technology was suitable for correlation analysis of symptoms and indexes in fatigue population, and tongue and pulse data had a certain diagnostic contribution to the classification of fatigue population.

Tongue color clustering and visual application based on 2D information.

PURPOSE: Studies have shown the association between tongue color and diseases. To help clinicians make more objective and accurate decisions quickly, we take unsupervised learning to deal with the basic clustering of tongue color in a 2D way. METHODS: A total of 595 typical tongue images were analyzed. The 3D information extracted from the image was transformed into 2D information by principal component analysis (PCA). K-Means was applied for clustering into four diagnostic groups. The results were evaluated by clustering accuracy (CA), Jaccard similarity coefficient (JSC), and adjusted rand index (ARI). RESULTS: The new 2D information totally retained 89.63% original information in the L*a*b* color space. And our methods successfully classified tongue images into four clusters and the CA, ARI, and JSC were 89.04%, 0.721, and 0.890, respectively. CONCLUSIONS: The 2D information of tongue color can be used for clustering and to improve the visualization. K-Means combined with PCA could be used for tongue color classification and diagnosis. Methods in the paper might provide reference for the other research based on image diagnosis technology.

A Study of Machine-Learning Classifiers for Hypertension Based on Radial Pulse Wave.

OBJECTIVE: In this study, machine learning was utilized to classify and predict pulse wave of hypertensive group and healthy group and assess the risk of hypertension by observing the dynamic change of the pulse wave and provide an objective reference for clinical application of pulse diagnosis in traditional Chinese medicine (TCM). METHOD: The basic information from 450 hypertensive cases and 479 healthy cases was collected by self-developed H20 questionnaires and pulse wave information was acquired by self-developed pulse diagnostic instrument (PDA-1). H20 questionnaires and pulse wave information were used as input variables to obtain different machine learning classification models of hypertension. This method was aimed at analyzing the influence of pulse wave on the accuracy and stability of machine learning model, as well as the feature contribution of hypertension model after removing noise by K-means. RESULT: Compared with the classification results before removing noise, the accuracy and the area under the curve (AUC) had been improved. The accuracy rates of AdaBoost, Gradient Boosting, and Random Forest (RF) were 86.41%, 86.41%, and 85.33%, respectively. AUC were 0.86, 0.86, and 0.85, respectively. The maximum accuracy of SVM increased from 79.57% to 83.15%, and the AUC stability increased from 0.79 to 0.83. In addition, the features of importance on traditional statistics and machine learning were consistent. After removing noise, the features with large changes were h1/t1, w1/t, t, w2, h2, t1, and t5 in AdaBoost and Gradient Boosting (top10). The common variables for machine learning and traditional statistics were h1/t1, h5, t, Ad, BMI, and t2. CONCLUSION: Pulse wave-based diagnostic method of hypertension has significant value in reference. In view of the feasibility of digital-pulse-wave diagnosis and dynamically evaluating hypertension, it provides the research direction and foundation for Chinese medicine in the dynamic evaluation of modern disease diagnosis and curative effect.

Tongue Image Database Construction Based on the Expert Opinions: Assessment for Individual Agreement and Methods for Expert Selection.

This study aims at introducing a method for individual agreement evaluation to identify the discordant raters from the experts' group. We exclude those experts and decide the best experts selection method, so as to improve the reliability of the constructed tongue image database based on experts' opinions. Fifty experienced experts from the TCM diagnostic field all over China were invited to give ratings for 300 randomly selected tongue images. Gwet's AC1 (first-order agreement coefficient) was used to calculate the interrater and intrarater agreement. The optimization of the interrater agreement and the disagreement score were put forward to evaluate the external consistency for individual expert. The proposed method could successfully optimize the interrater agreement. By comparing three experts' selection methods, the interrater agreement was, respectively, increased from 0.53 [0.32-0.75] for original one to 0.64 [0.39-0.80] using method A (inclusion of experts whose intrarater agreement>0.6), 0.69 [0.63-0.81] using method B (inclusion of experts whose disagreement score="0"), and 0.76 [0.67-0.83] using method C (inclusion of experts whose intrarater agreement>0.6& disagreement score="0"). In this study, we provide an estimate of external consistency for individual expert, and the comprehensive consideration of both the internal consistency and the external consistency for each expert would be superior to either one in the tongue image construction based on expert opinions.

Clinical epidemiology of gastric cancer in Hehuang valley of China: a 10-year epidemiological study of gastric cancer.

AIM: To investigate the clinical epidemiological characteristics of gastric cancer in the Hehuang valley, China, to provide a reference for treatment and prevention of regional gastric cancer. METHODS: Between February 2003 and February 2013, the records of 2419 patients with gastric cancer were included in this study. The patient's characteristics, histological and pathological features, as well as the dietary habits of the patients, were investigated. RESULTS: The clinical data showed that adenocarcinoma was the leading histological type of gastric cancer in this area. Characteristics of gastric cancer in different ethnic groups and age showed that the 60.55-65.50 years group showed the high incidence of gastric cancer in all ethnic groups. There were more male gastric cancer patients than female. Intestinal was the most common type of gastric cancer in the Hehuang valley. There was no significant difference in the proportion of sex in terms of Helicobacter pylori infection. The impact of dietary habits on gastric cancer showed that regular consumption of fried or grilled food, consumption of high-salt, high-fat and spicy food and drinking strong Boiled brick-tea were three important factors associated with gastric cancer in males and females. CONCLUSION: Differences existed in race, sex, and age of patients according to the epidemiology of gastric cancer in the Hehuang valley. Moreover, dietary habits was also an important factor contributing to gastric cancer.