Oral epithelial dysplasia detection and grading in oral leukoplakia using deep learning.

BACKGROUND: The grading of oral epithelial dysplasia is often time-consuming for oral pathologists and the results are poorly reproducible between observers. In this study, we aimed to establish an objective, accurate and useful detection and grading system for oral epithelial dysplasia in the whole-slides of oral leukoplakia. METHODS: Four convolutional neural networks were compared using the image patches from 56 whole-slide of oral leukoplakia labeled by pathologists as the gold standard. Sequentially, feature detection models were trained, validated and tested with 1,000 image patches using the optimal network. Lastly, a comprehensive system named E-MOD-plus was established by combining feature detection models and a multiclass logistic model. RESULTS: EfficientNet-B0 was selected as the optimal network to build feature detection models. In the internal dataset of whole-slide images, the prediction accuracy of E-MOD-plus was 81.3% (95% confidence interval: 71.4-90.5%) and the area under the receiver operating characteristic curve was 0.793 (95% confidence interval: 0.650 to 0.925); in the external dataset of 229 tissue microarray images, the prediction accuracy was 86.5% (95% confidence interval: 82.4-90.0%) and the area under the receiver operating characteristic curve was 0.669 (95% confidence interval: 0.496 to 0.843). CONCLUSIONS: E-MOD-plus was objective and accurate in the detection of pathological features as well as the grading of oral epithelial dysplasia, and had potential to assist pathologists in clinical practice.

The clinical implication and translational research of OSCC differentiation.

BACKGROUND: The clinical value and molecular characteristics of tumor differentiation in oral squamous cell carcinoma (OSCC) remain unclear. There is a lack of a related molecular classification prediction system based on pathological images for precision medicine. METHODS: Integration of epidemiology, genomics, experiments, and deep learning to clarify the clinical value and molecular characteristics, and develop a novel OSCC molecular classification prediction system. RESULTS: Large-scale epidemiology data (n = 118,817) demonstrated OSCC differentiation was a significant prognosis indicator (p < 0.001), and well-differentiated OSCC was more chemo-resistant than poorly differentiated OSCC. These results were confirmed in the TCGA database and in vitro. Furthermore, we found chemo-resistant related pathways and cell cycle-related pathways were up-regulated in well- and poorly differentiated OSCC, respectively. Based on the characteristics of OSCC differentiation, a molecular grade of OSCC was obtained and combined with pathological images to establish a novel prediction system through deep learning, named ShuffleNetV2-based Molecular Grade of OSCC (SMGO). Importantly, our independent multi-center cohort of OSCC (n = 340) confirmed the high accuracy of SMGO. CONCLUSIONS: OSCC differentiation was a significant indicator of prognosis and chemotherapy selection. Importantly, SMGO could be an indispensable reference for OSCC differentiation and assist the decision-making of chemotherapy.

Periodontitis and the risk of all-cause and cause-specific mortality among US adults with diabetes: A population-based cohort study.

AIM: To evaluate the association between periodontitis, all-cause and cause-specific mortality, and its prognostic utility among adults with diabetes. MATERIALS AND METHODS: Periodontal health records were retrieved from the NHANES database for 4297 participants with diabetes aged >30 years at baseline during 1988-1994, 1999-2004 and 2009-2014. Multivariable Cox proportional hazards regression model was applied to calculate the hazard ratios (HRs) and 95% confidence intervals (CIs) for moderate/severe periodontitis with all-cause and cause-specific mortality in participants with diabetes. Area under the curve (AUC) was used to assess predictive value. RESULTS: During a median follow-up of 15.41 years, 1701 deaths occurred. After multivariate adjustments, moderate/severe periodontitis was significantly associated with increased risk of all-cause (HR: 1.27; 95% CI: 1.07-1.50; p = .005) and cardiovascular disease (CVD)-related (HR: 1.35, 95% CI: 1.03-1.76, p = .031) mortality in participants with diabetes. The absolute risk difference based on the cumulative incidence information was 0.022 (5-year, 95% CI: 0.021-0.023) and 0.044 (10-year, 95% CI: 0.041-0.048). Periodontitis improved the prediction of all-cause (AUC: 0.652; 95% CI: 0.627-0.676) and CVD-related (AUC: 0.649; 95% CI: 0.624-0.676) mortality over standard risk factors (all-cause: AUC: 0.631; 95% CI: 0.606-0.656; CVD-related: AUC: 0.629; 95% CI: 0.604-0.655). CONCLUSIONS: Moderate/severe periodontitis is associated with an increased risk of all-cause and CVD-related mortality in adults with diabetes. Periodontitis might represent a marker for residual risk.

CD8+ tissue-resident memory T cells induce oral lichen planus erosion via cytokine network.

CD8+ tissue-resident memory T (CD8+ Trm) cells play key roles in many immune-inflammation-related diseases. However, their characteristics in the pathological process of oral lichen planus (OLP) remains unclear. Therefore, we investigated the function of CD8+ Trm cells in the process of OLP. By using single-cell RNA sequencing profiling and spatial transcriptomics, we revealed that CD8+ Trm cells were predominantly located in the lamina propria adjacent to the basement membrane and were significantly increased in patients with erosive oral lichen planus (EOLP) compared to those with non-erosive oral lichen planus (NEOLP). Furthermore, these cells displayed enhanced cytokine production, including IFN-γ (Interferon-gamma, a pro-inflammatory signaling molecule), TNF-α (Tumor Necrosis Factor-alpha, a cytokine regulating inflammation), and IL-17 (Interleukin-17, a cytokine involved in immune response modulation), in patients with EOLP. And our clinical cohort of 1-year follow-up was also supported the above results in RNA level and protein level. In conclusion, our study provided a novel molecular mechanism for triggering OLP erosion by CD8+ Trm cells to secrete multiple cytokines, and new insight into the pathological development of OLP.

SNAI2 promotes the malignant transformation of oral leukoplakia by modulating p-EMT.

OBJECTIVE: Snail family transcriptional repressor 2 (SNAI2) is a key regulator of partial epithelial-mesenchymal transition (p-EMT) and is associated with tumorigenesis. Whether SNAI2 promotes oral leukoplakia (OLK) malignant transformation by modulating p-EMT is unclear. MATERIALS AND METHODS: This study utilized two clinical datasets (GSE26549 and GSE85195) from the Gene Expression Omnibus database, cytological experiments, and a 4-nitroquinoline 1-oxide-induced mice model to explore the role of SNAI2 in OLK malignant transformation. RESULTS: The clinical cohort found SNAI2, as a risk factor (HR = 2.50, 95% CI: 1.08-5.79, p = 0.033), could promote OLK malignant transformation (p = 0.012). Cytological experiments indicated that SNAI2 overexpression promoted DOK cell proliferation, invasion, migration, and increase the protein expression of p-EMT relative signatures, whereas SNAI2 silencing has opposite effects. Furthermore, the mice model and clinical datasets demonstrated the expression of SNAI2 and p-EMT relative signatures were increased with OLK malignant transformation. And SNAI2 was strongly correlated with p-EMT. Besides, co-expressed genes of SNAI2 were also enriched in p-EMT relative biological processes and signaling pathways. CONCLUSIONS: p-EMT plays a significant role in promoting the OLK malignant transformation. As an important regulator of p-EMT, SNAI2 could be a target to block the OLK malignant transformation.

The interleukin-6 family in tissues is closely related to the clinical outcomes of oral lichen planus.

OBJECTIVE: We attempted to investigate the role of interleukin-6 (IL-6) family expression in local tissues as it relates to presentations and outcomes in oral lichen planus (OLP), which is a common chronic inflammatory oral disease. MATERIALS AND METHODS: A clinical follow-up cohort of OLP patients was established, and a biological sample library was constructed with categorization into erosive type (EOLP) and nonerosive type (NEOLP). Transcriptome sequencing of the lesions was then performed. A multiple regression model was used to explore the differences in IL-6 family expression among patients with different clinical types and clinical outcomes. RESULTS: OLP tissue transcriptome sequencing showed that IL-6 family expression in EOLP increased significantly. It was also found that IL-6 family factors in the OLP recurrent erosion group were significantly increased compared to the persistent nonerosion group. Based on the multiple regression analysis of the OLP clinical cohort, it was found that the increased expression of the IL-6 family was closely related to the clinical types and clinical outcomes of OLP. CONCLUSION: The high expression of the IL-6 family is closely related to the erosion of local mucosa and poor prognosis of OLP patients. IL-6-related factors may be used as therapeutic targets for OLP patients.

High-Accuracy Oral Squamous Cell Carcinoma Auxiliary Diagnosis System Based on EfficientNet.

It is important to diagnose the grade of oral squamous cell carcinoma (OSCC), but the current evaluation of the biopsy slide still mainly depends on the manual operation of pathologists. The workload of manual evaluation is large, and the results are greatly affected by the subjectivity of the pathologists. In recent years, with the development and application of deep learning, automatic evaluation of biopsy slides is gradually being applied to medical diagnoses, and it has shown good results. Therefore, a new OSCC auxiliary diagnostic system was proposed to automatically and accurately evaluate the patients' tissue slides. This is the first study that compared the effects of different resolutions on the results. The OSCC tissue slides from The Cancer Genome Atlas (TCGA, n=697) and our independent datasets (n=337) were used for model training and verification. In the test dataset of tiles, the accuracy was 93.1% at 20x resolution (n=306,134), which was higher than that at 10x (n=154,148, accuracy=90.9%) and at 40x (n=890,681, accuracy=89.3%). The accuracy of the new system based on EfficientNet, which was used to evaluate the tumor grade of the biopsy slide, reached 98.1% [95% confidence interval (CI): 97.1% to 99.1%], and the area under the receiver operating characteristic curve (AUROC) reached 0.998 (95%CI: 0.995 to 1.000) in the TCGA dataset. When verifying the model on the independent image dataset, the accuracy still reached 91.4% (95% CI: 88.4% to 94.4%, at 20x) and the AUROC reached 0.992 (95%CI: 0.982 to 1.000). It may benefit oral pathologists by reducing certain repetitive and time-consuming tasks, improving the efficiency of diagnosis, and facilitating the further development of computational histopathology.

Abnormal Micronutrient Intake Is Associated with the Risk of Periodontitis: A Dose-response Association Study Based on NHANES 2009-2014.

The association between micronutrient intake and the risk of periodontitis has received much attention in recent years. However, most studies focused on the linear relationship between them. This study aimed to explore the dose-response association between micronutrient intake and periodontitis. A total of 8959 participants who underwent a periodontal examination, and reported their micronutrient intake levels were derived from the US National Health and Nutrition Examination Survey (NHANES, 2009-2014) database. Logistic regression was performed to evaluate associations between micronutrient intake and periodontitis after propensity score matching (PSM), and restricted cubic splines (RCS) analysis was conducted to explore the dose-response associations. Following PSM, 5530 participants were included in the RCS analysis. The risk of periodontitis was reduced with sufficient intake of the following micronutrients: vitamin A, vitamin B1, vitamin B2, and vitamin E. In addition, the risk of periodontitis was increased with excessive intake of the following micronutrients: vitamin B1 (1.8 mg/day, males; 1.3 mg/day, females), vitamin C (90 mg/day, males), and copper (1.1 mg/day, combined). In conclusion, a linear association was found between vitamin A, vitamin B2, vitamin C, and copper and periodontitis-namely, a sufficient intake of vitamin A and vitamin B2 might help reduce the prevalence of periodontitis; by contrast, a high intake of vitamin C and copper increased the risk. In addition, a nonlinear dose-response association was found for the incidence of periodontitis with vitamin B1 and vitamin E. When within reasonable limits, supplemental intake helped reduce the prevalence of periodontitis, while excessive intake did not help significantly and might even increase the risk. However, confounding factors, such as health awareness, should still be considered.

CircRFWD3 promotes HNSCC metastasis by modulating miR-27a/b/PPARγ signaling.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer in the world, the 5-year survival rate of patients with HNSCC is still about 50% due to frequent metastasis and recurrence. Circular RNAs (circRNAs) have been characterized as key regulators of gene expression in numerous malignancies. However, the role of circRNA in HNSCC metastasis remains largely unknown. Here, we demonstrated that the circRFWD3 was significantly upregulated in HNSCC tissues and cell lines by circRNA microarray analysis and qPCR. Notably, high expression of circRFWD3 is related to highly aggressive HNSCC cell lines and lymph node metastasis in HNSCC patients. After that, Sanger sequencing, RNase R, and actinomycin D assay were performed to verify the ring structure of circRFWD3. Then functional experiments found it could promote the metastasis of HNSCC cells both in vitro and in vivo. Mechanistically, a dual-luciferase reporter assay, FISH, RIP, RNA pull-down, RNA-seq, and western blot experiments were employed and found that circRFWD3 served as a miRNAs sponge for miR-27a/27b, leading to the upregulation of PPARγ, and then promoted HNSCC metastasis via NF-κB/MMP13 pathway. Finally, ISH and IHC were carried out to determine the expression levels and clinical significances of circRFWD3 and PPARγ in clinical cohorts of HNSCC. According to the analysis results from two independent HNSCC cohorts, upregulated expression of circRFWD3 and PPARγ were positively associated with worse survival in patients with HNSCC. Overall, our results uncover that circRFWD3 acts a critical role in promoting the aggressiveness of HNSCC cells and is a prognostic marker for the disease, indicating that circRFWD3 may act as a potential therapeutic target in HNSCC.

Effect of Marital Status on Upper Digestive Tract Tumor Survival: Married Male Patients Exhibited a Better Prognosis.

Purpose: Marital status has been associated with the outcomes in several types of cancer, but less is known about upper digestive tract tumors (UDTTs). The study aims to explore the effect of marital status on the survival outcomes of UDTT. Methods: We collected patient cases of UDTT using the Surveillance, Epidemiology, and End Results (SEER) database between 1975 and 2016. The univariate analyses of overall survival (OS) and cancer-specific survival (CSS) were performed using the Kaplan-Meier method. The multivariate survival analyses were performed using Cox proportional hazard model. Results: A total of 282,189 patients were included, with 56.42, 16.30, 13.33, and 13.95% of patients married, never married, divorced or separated, and widowed, respectively. The significant differences were observed among married, never-married, divorced or separated, and widowed patients with regard to the year of diagnosis, sex, age, race, pathological type, anatomical site, the number of primary tumor, grade, rate of surgery performed, radiotherapy, chemotherapy (p < 0.001). The proportions of patients with 3-year and 5-year OS were 54.22 and 48.02% in the married group, 46.96 and 41.12% in the never-married group, 44.24 and 38.06% in the divorced or separated group, 34.59 and 27.57% in the widowed group, respectively (p < 0.001); the proportions of patients with 3-year and 5-year CSS were 70.76 and 68.13% in the married group, 62.44 and 59,93% in the never-married group, 63.13 and 60.53% in the divorced or separated group, 62.11 and 58.89% in the widowed group, respectively (p < 0.001); all these data indicated married patients exhibited favorable OS and CSS than never-married, divorced or separated, and widowed patients. Men in the married group showed better OS (HR, 1.16; 95%CI: 1.11-1.22) and CSS (HR, 0.96; 95%CI: 0.92-1.23) than those in the never-married group. Conclusion: This study reveals that marital status is an independent prognostic factor for OS and CSS of patients with UDTT. Married male patients with UDTT trend to have a better prognosis.

Anxiety and oral lichen planus.

OBJECTIVES: To investigate the association between oral lichen planu(OLP) and anxiety. SUBJECTS AND METHODS: This study included 174 OLP patients and 174 healthy controls. We assessed anxiety by Self-rating Anxiety Scale (SAS) and recorded OLP lesion type and severity. t test and analysis of variance were applied for continuous variants and chi-square test was performed for categorical variants. Multiple linear regression and logistic regression analysis were used for multi-variable analysis. RESULTS: he SAS score of OLP patients was higher than that of healthy individuals. There was no significant difference in SAS score between the OLP subgroups, obtained according to age, type, and severity, respectively. Multiple linear regression analysis showed gender was the only factor that affected the SAS score of OLP patients. Compared with weakly anxiety-related groups, the SAS score and female ratio of highly anxiety-related group were obviously higher. Logistic regression analysis demonstrated that males were less exposed to highly anxiety-related types than females. CONCLUSIONS: OLP patients tend to be more anxious compared with healthy individuals, and female patients are more anxious than male patients. There might be two types of OLP patients: weakly anxiety-related or highly anxiety-related. These results highlight the significance of psychological counseling in OLP disease management.

Association of Human Papillomavirus With Oral Lichen Planus and Oral Leukoplakia: A Meta-analysis.

OBJECTIVES: To estimate the association of human papillomavirus (HPV) infection with oral lichen planus (OLP) and oral leukoplakia (OLK), and determine risk cofactors. STUDY DESIGN: Seven databases were searched for case-control or cross-sectional studies of OLP and OLK with healthy controls, published between 1976 and 2020. The Meta package of R software was applied to calculate the pooled odds ratios (OR) and 95% confidence interval (CI). RESULTS: Thirty-six articles were finally included. OLP and OLK cases had a higher association with HPV infection than controls (OLP: OR: 4.91, 95% CI: 2.76-8.72; OLK: OR: 2.51, 95% CI: 1.55-4.07). In subgroup analyses, the OR of HPV infection was higher with erosive lesions than with nonerosive lesions (OLP: OR: 5.36 and 3.47, respectively; OLK: OR: 3.34 and 3.21, respectively). Oral lesions were more strongly associated with HPV16/18 than with HPV6/11 (OLP: OR: 7.84 and 1.42, respectively; OLK: OR: 6.05 and 1.87, respectively) and varied by geographic region (OLP: OR: 4.01-7.02; OLK: OR: 1.46-27.13). CONCLUSIONS: Oral HPV infection, particularly infection with HPV 16/18, was strongly associated with OLP and OLK. Risk cofactors included erosive lesions and geographic region.

Low-level laser therapy in the prevention and treatment of oral mucositis: a systematic review and meta-analysis.

OBJECTIVE: The aim of this study was to determine whether prophylactic and therapeutic low-level laser therapy (LLLT), compared with placebo or no therapy, reduced the risk of severe oral mucositis (OM) in patients receiving chemotherapy or radiotherapy. STUDY DESIGN: We searched for articles published on randomized controlled trials (RCTs) in the databases MEDLINE, EMBASE, Cochrane Library, Cochrane Central Register of Controlled Trials, Web of Science, and Clinical Trials, until December 2018. RCTs were filtered on the basis of eligibility criteria, and data were analyzed by using R software 3.5.2. RESULTS: Overall, 30 studies were included in the meta-analysis. Prophylactic LLLT reduced the overall risk of severe OM (relative risk [RR] = 0.40; 95% confidence interval [CI]: 0.28-0.57; P < .01). Therapeutic LLLT substantially reduced the duration of severe OM (P < .01). LLLT also reduced the overall mean grade of OM, overall incidence of severe pain, mean score of pain, and incidence of severe OM, at the most anticipated time. CONCLUSIONS: Prophylactic and therapeutic LLLT can reduce the risk of severe OM in patients receiving chemotherapy or radiotherapy.

High expression of ACE2 receptor of 2019-nCoV on the epithelial cells of oral mucosa.

It has been reported that ACE2 is the main host cell receptor of 2019-nCoV and plays a crucial role in the entry of virus into the cell to cause the final infection. To investigate the potential route of 2019-nCov infection on the mucosa of oral cavity, bulk RNA-seq profiles from two public databases including The Cancer Genome Atlas (TCGA) and Functional Annotation of The Mammalian Genome Cap Analysis of Gene Expression (FANTOM5 CAGE) dataset were collected. RNA-seq profiling data of 13 organ types with para-carcinoma normal tissues from TCGA and 14 organ types with normal tissues from FANTOM5 CAGE were analyzed in order to explore and validate the expression of ACE2 on the mucosa of oral cavity. Further, single-cell transcriptomes from an independent data generated in-house were used to identify and confirm the ACE2-expressing cell composition and proportion in oral cavity. The results demonstrated that the ACE2 expressed on the mucosa of oral cavity. Interestingly, this receptor was highly enriched in epithelial cells of tongue. Preliminarily, those findings have explained the basic mechanism that the oral cavity is a potentially high risk for 2019-nCoV infectious susceptibility and provided a piece of evidence for the future prevention strategy in dental clinical practice as well as daily life.