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.

Angelicin inhibits cell growth and promotes apoptosis in oral squamous cell carcinoma by negatively regulating DUSP6/cMYC signaling pathway.

Angelicin has been reported to have antitumor effects on many types of cancer. However, few studies on angelicin in oral squamous cell carcinoma (OSCC) have been performed. We performed cell cycle and apoptosis analyses to assess the effect of angelicin on OSCC cells. We conducted RNA-seq studies to reveal differentially expressed genes (DEGs). Dual-specificity phosphatase 6 (DUSP6) and c-MYC were strongly down-regulated differential genes. Silencing RNA (siRNA) was used to knockdown DUSP6. The mouse xenograft model was used to mimic OSCC. Angelicin inhibited OSCC in vitro. We found that DUSP6 interacted with c-MYC. DUSP6 knockdown group and DUSP6 knockdown + angelicin group had similar effects of OSCC cells. Angelicin could reduce tumor formation, DUSP6, and c-MYC expression in vivo. Compared with paclitaxel, the tumor inhibition effect of the two drugs was similar. However, angelicin did not cause weight loss and had lower toxicity. In sum, Angelicin has antitumor effects on OSCC in vitro and vivo by negatively regulating the DUSP6 mediated c-MYC signaling pathway.

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.

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.

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.

Humoral immune disorders affect clinical outcomes of oral lichen planus.

OBJECTIVES: The molecular characteristics of oral lichen planus (OLP) are still unclear, and it is not possible to distinguish the clinical outcome of OLP patients in a short period of time for follow-up. Here, we investigate the molecular characteristics of lesions in patients with stable lichen planus (SOLP) and recalcitrant erosive oral lichen planus (REOLP). METHODS: Our clinical follow-up cohort was split into SOLP and REOLP groups based on the follow-up clinical data. The core modules associated with the clinical information were identified by weighted gene co-expression network analysis (WGCNA). The OLP cohort samples were divided into two groups by molecular typing, and a prediction model for OLP was created by training neural networks with the neuralnet package. RESULTS: We screened 546 genes in five modules. After doing a molecular type of OLP, it was determined that B cells might have a significant impact on the clinical outcome of OLP. In addition, by means of machine learning, a prediction model was developed to predict the clinical regression of OLP with greater accuracy than the existing clinical diagnostic. CONCLUSIONS: Our study revealed humoral immune disorders may make an important contribution to the clinical outcome of OLP.

Hydroxychloroquine is effective in oral lichen planus: A multicenter, randomized, controlled trial.

OBJECTIVES: This study was aimed to evaluate the safety and benefit of short-term application of hydroxychloroquine in the management of atrophic/erosive/ulcerative oral lichen planus (OLP). METHODS: This multicenter, randomized, controlled, evaluator-blinded, prospective clinical trial was performed from October 1, 2019, to September 1, 2022. A total of 99 patients were randomized to receive systemic use of hydroxychloroquine (n = 50), or topical use of 0.05% dexamethasone (n = 49) for 4 weeks. The response to both treatment modalities was evaluated according to reticulation, hyperemic, and ulceration (RHU) score and visual analog scale (VAS) score. RESULTS: After 4 weeks of medication, both groups showed substantial reduction in RHU and VAS score (p < 0.05). In hydroxychloroquine group, the average of RHU score was reduced from 10.60 to 7.68 (dropped 27.49%), and the average of VAS score was reduced from 3.74 to 2.47 (dropped 34.09%). There were no differences between the two groups in reduction of RHU score and VAS score (p > 0.05). Single factor analysis found hyperemic area (p = 0.019) and erosive/ulcerative area (p = 0.024) had impacts on drug efficacy of hydroxychloroquine, and logistic regression revealed that no factors (p > 0.05) influenced its efficacy. CONCLUSION: These findings indicate hydroxychloroquine is a safe and effective agent in treating atrophic/erosive/ulcerative OLP.

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.

[Establishment of Ace2 knockout mouse model with CRISPR/Cas9 gene targeting technology].

The aim of the study was to establish Ace2 (angiotensin-converting enzyme 2) knockout mouse model with CRISPR/Cas9 gene targeting technology. A vector targeting Ace2 gene knockout was constructed with the primers of single-guide RNA (gRNA), and then transcribed gRNA/Cas9 mRNA was micro-injected into the mouse zygote. The deletion of exons 3 to 18 of Ace2 gene in mice was detected and identified by PCR and gene sequencing. The Ace2 gene knock-out mice were bred and copulated. Ace2 protein and mRNA expression were detected by Western blot and qRT-PCR in F3 progeny knock-out male mice. The gRNA expression vector was successfully constructed and transcribed in vitro, and active gRNA and Cas9 mRNA were injected directly into zygote. The deletion of exons 3 to 18 of Ace2 gene in six positive founder mice as the F0 generation were confirmed by PCR and gene sequencing. Six founder mice were mated with wild-type mice, then achieved F1 generation were mated and produced F2 generation. The female positive mouse of F2 was selected to mate with wild-type mice and produce Ace2-/Y mice of F3 generation. Ace2 mRNA and protein were not detected in tissues of these Ace2-/Y mice. In conclusion, a mouse model with Ace2 deficiency has been successfully established with CRISPR/Cas9 technique, which shall lay a foundation for future investigation of Ace2.

Dietary curcumin ameliorates aging-related cerebrovascular dysfunction through the AMPK/uncoupling protein 2 pathway.

BACKGROUND/AIMS: Age-related cerebrovascular dysfunction contributes to stroke, cerebral amyloid angiopathy, cognitive decline and neurodegenerative diseases. One pathogenic mechanism underlying this effect is increased oxidative stress. Up-regulation of mitochondrial uncoupling protein 2 (UCP2) plays a crucial role in regulating reactive oxygen species (ROS) production. Dietary patterns are widely recognized as contributors to cardiovascular and cerebrovascular disease. In this study, we tested the hypothesis that dietary curcumin, which has an antioxidant effect, can improve aging-related cerebrovascular dysfunction via UCP2 up-regulation. METHODS: The 24-month-old male rodents used in this study, including male Sprague Dawley (SD) rats and UCP2 knockout (UCP2-/-) and matched wild type mice, were given dietary curcumin (0.2%). The young control rodents were 6-month-old. Rodent cerebral artery vasorelaxation was detected by wire myograph. The AMPK/UCP2 pathway and p-eNOS in cerebrovascular and endothelial cells were observed by immunoblotting. RESULTS: Dietary curcumin administration for one month remarkably restored the impaired cerebrovascular endothelium-dependent vasorelaxation in aging SD rats. In cerebral arteries from aging SD rats and cultured endothelial cells, curcumin promoted eNOS and AMPK phosphorylation, up-regulated UCP2 and reduced ROS production. These effects of curcumin were abolished by either AMPK or UCP2 inhibition. Chronic dietary curcumin significantly reduced ROS production and improved cerebrovascular endothelium-dependent relaxation in aging wild type mice but not in aging UCP2-/- mice. CONCLUSIONS: Curcumin improves aging-related cerebrovascular dysfunction via the AMPK/UCP2 pathway.

Cardiac protective effects of irbesartan via the PPAR-gamma signaling pathway in angiotensin-converting enzyme 2-deficient mice.

BACKGROUND: Angiotensin-converting enzyme 2 (ACE2), a monocarboxypeptidase which metabolizes angiotensin II (Ang II) to generate Ang-(1-7), has been shown to prevent cardiac hypertrophy and injury but the mechanism remains elusive. Irbesartan has the dual actions of angiotensin receptor blockade and peroxisome proliferator-activated receptor-γ (PPARγ) activation. We hypothesized that irbesartan would exert its protective effects on ACE2 deficiency-mediated myocardial fibrosis and cardiac injury via the PPARγ signaling. METHODS: 10-week-old ACE2 knockout (ACE2KO; Ace2(-/y)) mice received daily with irbesartan (50 mg/kg) or saline for 2 weeks. The wild-type mice (Ace2(+/y)) were used to the normal controls. We examined changes in myocardial ultrastructure, fibrosis-related genes and pathological signaling by real-time PCR gene array, Western blotting, Masson trichrome staining and transmission electron microscope analyses, respectively. RESULTS: Compared with the Ace2(+/y) mice, cardiac expression of PPARα and PPARγ were reduced in Ace2(-/y) mice and the myocardial collagen volume fraction (CVF) and expression of fibrosis-related genes were increased, including transforming growth factor-ß1 (TGFß1), connective tissue growth factor (CTGF), collagen I and collagen III. Moreover, ACE2 deficiency triggered cardiac hypertrophy, increased myocardial fibrosis and adverse ultrastructure injury in ACE2KO hearts with higher levels of atrial natriuretic factor (ANF) and phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), without affecting cardiac systolic function. Intriguingly, treatment with irbesartan significantly reversed ACE2 deficiency-mediated pathological hypertrophy and myocardial fibrosis in Ace2(-/y) mice linked with enhancement of plasma Ang-(1-7) level and downregulation of AT1 receptor in heart. Consistent with attenuation of myocardial fibrosis and ultrastructure injury, the myocardial CVF and levels of ANF, TGFß1, CTGF, collagen I, collagen III and phosphorylated ERK1/2 were lower, and expression of PPARγ was higher in ACE2KO mice in response to irbesartan treatment, without affecting cardiac expression of PPARα, PPARδ, ß-myosin heavy chain, TGFß2 and fibronectin. CONCLUSIONS: We conclude that irbesartan prevents ACE2 deficiency-mediated pathological hypertrophy and myocardial fibrosis in ACE2 mutant mice via activation of the PPARγ signaling and suppression of the TGFß-CTGF-ERK signaling, resulting in attenuation of myocardial injury. Drugs targeting ACE2 and PPARγ represent potential candidates to prevent and treat myocardial injury and related cardiac disorders.

TRPV1-mediated UCP2 upregulation ameliorates hyperglycemia-induced endothelial dysfunction.

BACKGROUND: Diabetic cardiovascular complications are characterised by oxidative stress-induced endothelial dysfunction. Uncoupling protein 2 (UCP2) is a regulator of mitochondrial reactive oxygen species (ROS) generation and can antagonise oxidative stress, but approaches that enhance the activity of UCP2 to inhibit ROS are scarce. Our previous studies show that activation of transient receptor potential vanilloid 1 (TRPV1) by capsaicin can prevent cardiometabolic disorders. In this study, we conducted experiments in vitro and in vivo to investigate the effect of capsaicin treatment on endothelial UCP2 and oxidative stress. We hypothesised that TRPV1 activation by capsaicin attenuates hyperglycemia-induced endothelial dysfunction through a UCP2-mediated antioxidant effect. METHODS: TRPV1(-/-), UCP2(-/-) and db/db mice, as well as matched wild type (WT) control mice, were included in this study. Some mice were subjected to dietary capsaicin for 14 weeks. Arteries isolated from mice and endothelial cells were cultured. Endothelial function was examined, and immunohistological and molecular analyses were performed. RESULTS: Under high-glucose conditions, TRPV1 expression and protein kinase A (PKA) phosphorylation were found to be decreased in the cultured endothelial cells, and the effects of high-glucose on these molecules were reversed by the administration of capsaicin. Furthermore, high-glucose exposure increased ROS production and reduced nitric oxide (NO) levels both in endothelial cells and in arteries that were evaluated respectively by dihydroethidium (DHE) and DAF-2 DA fluorescence. Capsaicin administration decreased the production of ROS, restored high-glucose-induced endothelial dysfunction through the activation of TRPV1 and acted in a UCP2-dependent manner in vivo. Administration of dietary capsaicin for 14 weeks increased the levels of PKA phosphorylation and UCP2 expression, ameliorated the vascular oxidative stress and increased NO levels observed in diabetic mice. Prolonged dietary administration of capsaicin promoted endothelium-dependent relaxation in diabetic mice. However, the beneficial effect of capsaicin on vasorelaxation was absent in the aortas of UCP2(-/-) mice exposed to high-glucose levels. CONCLUSION: TRPV1 activation by capsaicin might protect against hyperglycemia-induced endothelial dysfunction through a mechanism involving the PKA/UCP2 pathway.

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.

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.

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.

Automatic Detection of Image-Based Features for Immunosuppressive Therapy Response Prediction in Oral Lichen Planus.

Oral lichen planus (OLP) is a chronic inflammatory disease, and the common management focuses on controlling inflammation with immunosuppressive therapy. While the response to the immunosuppressive therapy is heterogeneous, exploring the mechanism and prediction of the response gain greater importance. Here, we developed a workflow for prediction of immunosuppressive therapy response prediction in OLP, which could automatically acquire image-based features. First, 38 features were acquired from 208 OLP pathological images, and 6 features were subsequently obtained which had a significant impact on the effect of OLP immunosuppressive therapy. By observing microscopic structure and integrated with the corresponding transcriptome, the biological implications of the 6 features were uncovered. Though the pathway enrichment analysis, three image-based features which advantageous to therapy indicated the different lymphocytes infiltration, and the other three image-based features which bad for therapy respectively indicated the nicotinamide adenine dinucleotide (NADH) metabolic pathway, response to potassium ion pathway and adenosine monophosphate (AMP) activated protein kinase pathway. In addition, prediction models for the response to immunosuppressive therapy, were constructed with above image-based features. The best performance prediction model built by logistic regression showed an accuracy of 90% and the area under the receiver operating characteristic curve (AUROC) reached 0.947. This study provided a novel approach to automatically obtain biological meaningful image-based features from unannotated pathological images, which could indicate the immunosuppressive therapy in OLP. Besides, the novel and accurate prediction model may be useful for the OLP clinical management.

[Effects of ANP upon ion pump activity and gene expression in aortic smooth muscle cells from spontaneously hypertensive rats].

OBJECTIVE: To explore the effects of atrial natriuretic peptide (ANP) upon the activities of Na(+), K(+)-ATPase, Ca(2+)-ATPase and mRNA expression levels of Na(+), K(+)-ATPase alpha(1)-subunit and plasma membrane Ca(2+)-ATPase isoform 1 (PMCA1) in cultured thoracic aortic vascular smooth muscle cells (ASMCs) isolated from spontaneously hypertensive rats (SHR). METHODS: ASMCs isolated from 14-week-old male SHR and Wistar-Kyoto (WKY) rats were interference-cultured in different doses of ANP and Angiotensin II (AngII). The contents of ANP and AngII in supernatant from ASMCs were measured by radioimmunoassay. The activities of the above two ATPases were measured by biochemistry and enzymology. RT-PCR assay was employed to determine the relative levels of Na(+), K(+)-ATPase alpha(1)-subunit and PMCA1 mRNA in ASMCs. RESULTS: The ANP level of supernatant in SHR ASMCs was significantly lower than those from WKY control [(7.3 +/- 2.4) pg x 10(-6) cells vs (19.3 +/- 3.3) pg x 10(-6) cells, P < 0.01] while the content of AngII in SHR ASMCs was significantly higher than those from WKY control [(57 +/- 4) pg x 10(-6) cells vs (44 +/- 4) pg x 10(-6) cells, P < 0.01]. The activity of Na(+), K(+)-ATPase [(4.3 +/- 0.8) micromol x h(-1) x mg(-1) vs (5.3 +/- 1.0) micromol x h(-1) x mg(-1)], Ca(2+)-ATPase [(3.2 +/- 0.7) micromol x h(-1) x mg(-1) vs (4.5 +/- 0.7) micromol x h(-1) x mg(-1)] in ASMCs from SHR were significantly lower than those from WKY control (both P < 0.01). The mRNA expression of Na(+), K(+)-ATPase alpha(1)-subunit (0.524 +/- 0.025 vs 0.704 +/- 0.116), PMCA1 (0.193 +/- 0.030 vs 0.547 +/- 0.045) significantly decreased in ASMCs from SHR versus the WKY control (both P < 0.01). As compared with SHR control, exogenous ANP improved obviously the activities of Na(+), K(+)-ATPase, Ca(2+)-ATPase and expression of alpha(1)-subunit, PMCA1 mRNA in a does-dependent manner (P < 0.05-P < 0.01). Exogenous AngII (1 x 10(-9), 1 x 10(-8), 1 x 10(-7) mol/L) significantly repressed activities of Ca(2+)-ATPase and attenuated the expression of PMCA1 mRNA (P < 0.05-P < 0.01). Only AngII (1 x 10(-7) mol/L) significantly inhibited the activity of Na(+), K(+)-ATPase and attenuated the expression of Na(+), K(+)-ATPase alpha(1)-subunit mRNA (both P < 0.05). ANP antagonized the effects of AngII (1 x 10(-7) mol/L) upon the activities of two ATPases and the expression of Na(+), K(+)-ATPase alpha(1)-subunit PMCA1 mRNA (P < 0.05-P < 0.01). AngII (1 x 10(-7) mol/L) increased the Na(+), K(+)-ATPase activity and the expression of Na(+), K(+)-ATPase alpha(1)-subunit mRNA, repressed the Ca(2+)-ATPase activity and the expression of PMCA1 mRNA in ASMCs from WKY rat (P < 0.05-P < 0.01). ANP antagonized the effects of AngII (1 x 10(-7) mol/L) upon the activity of Ca(2+)-ATPase and the expression of PMCA1 mRNA (P < 0.05-P < 0.01), but did not antagonize the effects of AngII (1 x 10(-7) mol/L) upon the activity of Na(+), K(+)-ATPase and the expression of alpha(1)-subunit mRNA in ASMCs from WKY rats (P > 0.05). CONCLUSION: The decreased activities of Na(+), K(+)-ATPase and Ca(2+)-ATPase may be related to the abnormal autocrine of ANP and AngII in ASMC of SHR. ANP can antagonize the effects of AngII upon the activities of two ATPases and the expression of Na(+), K(+)-ATPase alpha(1)-subunit PMCA1 mRNA.

Establishment and characterization of a carcinoma-associated fibroblast cell line derived from a human salivary gland adenoid cystic carcinoma.

Salivary gland adenoid cystic carcinoma (SACC) is one of the most common malignancies in the oral and maxillofacial region. Carcinoma-associated fibroblast (CAF) is an important component in the tumor microenvironment and participates in SACC progression. In this study, we established a CAF cell line derived from a human SACC and named it CAF-SA. It was identified that CAF-SA expressed typical CAF biomarkers. Then, we studied the cellular communications between CAF-SA, tumor cells and endothelial cells. It was found that CAF-SA promoted the migration, invasion, and proliferation of SACC tumor cells in vitro. In addition, tube formation by endothelial cells was enhanced by CAF-SA. In vivo experiment showed that SACC cells formed larger xenografts in nude mice when they were transplanted with CAF-SA. Overall, we demonstrated that CAF-SA exhibited the most important defining feature of CAF by promoting cancer progression.

The sirtuin 6 prevents angiotensin II-mediated myocardial fibrosis and injury by targeting AMPK-ACE2 signaling.

Sirtuin 6 (SIRT6) is an important modulator of cardiovascular functions in health and diseases. However, the exact role of SIRT6 in heart disease is poorly defined. We hypothesized that SIRT6 is a negative regulator of angiotensin II (Ang II)-mediated myocardial remodeling, fibrosis and injury. The male Sprague-Dawley rats were randomized to Ang II (200 ng/kg/min) infusion with an osmotic minipump and pretreated with recombinant plasmids adeno-associated viral vector (AAV)-SIRT6 (pAAV-SIRT6) or pAAV-GFP for 4 weeks. Ang II triggered downregulated levels of SIRT6 and angiotensin-converting enzyme 2 (ACE2) and upregulated expression of connective tissue growth factor (CTGF) and proinflammatory chemokine fractalkine (FKN), contributing to enhanced cardiac fibrosis and ultrastructural injury. Reduced levels of phosphorylated pAMPK-α, increased myocardial hypertrophy and impaired heart dysfunction were observed in both Ang II-induced hypertensive rats and ACE2 knockout rats, characterized with increases in heart weight and left ventricular (LV) posterior wall thickness and decreases in LV ejection fraction and LV fractional shortening. More importantly, pAAV-SIRT6 treatment strikingly potentiated cardiac levels of pAMPKα and ACE2 as well as decreased levels of CTGF, FKN, TGFß1, collagen I and collagen III, resulting in alleviation of Ang II-induced pathological hypertrophy, myocardial fibrosis, cardiac dysfunction and ultrastructural injury in hypertensive rats. In conclusion, our findings confirmed cardioprotective effects of SIRT6 on pathological remodeling, fibrosis and myocardial injury through activation of AMPK-ACE2 signaling and suppression of CTGF-FKN pathway, indicating that SIRT6 functions as a partial agonist of ACE2 and targeting SIRT6 has potential therapeutic importance for cardiac fibrosis and heart disease.