circMTO1/miR-30c-5p/SOCS3 axis alleviates oral submucous fibrosis through inhibiting fibroblast-myofibroblast transition.

BACKGROUND: circRNAs have been shown to participate in diverse diseases; however, their role in oral submucous fibrosis (OSF), a potentially malignant disorder, remains obscure. Our preliminary experiments detected the expression of circRNA mitochondrial translation optimization 1 homologue (circMTO1) in OSF tissues (n = 20) and normal mucosa tissues (n = 20) collected from Hunan Xiangya Stomatological Hospital, and a significant decrease of circMTO1 expression was showed in OSF tissues. Therefore, we further explored circMTO1 expression in OSF. METHODS: Target molecule expression was detected using RT-qPCR and western blotting. The migration and invasion of buccal mucosal fibroblasts (BMFs) were assessed using wound healing and Transwell assays. The interaction between miR-30c-5p, circMTO1, and SOCS3 was evaluated using dual luciferase, RNA immunoprecipitation (RIP), and RNA pull-down assays. The colocalisation of circMTO1 and miR-30c-5p was observed using fluorescence in situ hybridisation (FISH). RESULTS: circMTO1 and SOCS3 expression decreased, whereas miR-30c-5p expression increased in patients with OSF and arecoline-stimulated BMFs. Overexpression of circMTO1 effectively restrained the fibroblast-myofibroblast transition (FMT), as evidenced by the increase in expression of Coll I, α-SMA, Vimentin, and the weakened migration and invasion functions in BMFs. Mechanistic studies have shown that circMTO1 suppresses FMT by enhancing SOCS3 expression by sponging miR-30c-5p and subsequently inactivating the FAK/PI3K/AKT pathway. FMT induced by SOCS3 silencing was reversed by the FAK inhibitor TAE226 or the PI3K inhibitor LY294002. CONCLUSION: circMTO1/miR-30c-5p/SOCS3 axis regulates FMT in arecoline-treated BMFs via the FAK/PI3K/AKT pathway. Expanding the sample size and in vivo validation could further elucidate their potential as therapeutic targets for OSF.

Identification of cuproptosis-related lncRNAs with the significance in prognosis and immunotherapy of oral squamous cell carcinoma.

Cuproptosis, a recently characterized programmed cell death mechanism, has emerged as a potential contributor to tumorigenesis, metastasis, and immune modulation. Long non-coding RNAs (lncRNAs) have demonstrated diverse regulatory roles in cancer and hold promise as biomarkers. However, the involvement and prognostic significance of cuproptosis-related lncRNAs (CRLs) in oral squamous cell carcinoma (OSCC) remain poorly understood. Based on TCGA-OSCC data, we integrated single-sample gene set enrichment analysis (ssGSEA), the LASSO algorithm, and the tumor immune dysfunction and exclusion (TIDE) algorithm. We identified 11 CRLs through differential expression, Spearman correlation, and univariate Cox regression analyses. Two distinct CRL-related subtypes were unveiled, delineating divergent survival patterns, tumor microenvironments (TME), and mutation profiles. A robust CRL-based signature (including AC107027.3, AC008011.2, MYOSLID, AC005785.1, AC019080.5, AC020558.2, AC025265.1, FAM27E3, and LINC02367) prognosticated OSCC outcomes, immunotherapy responses, and anti-tumor strategies. Superior predictive power compared to other lncRNA models was demonstrated. Functional assessments confirmed the influence of FAM27E3, LINC02367, and MYOSLID knockdown on OSCC cell behaviors. Remarkably, the CRLs-based signature maintained stability across OSCC patient subgroups, underscoring its clinical potential for survival prediction. This study elucidates CRLs' roles in TME of OSCC and establishes a potential signature for precision therapy.

Single-cell and bulk RNA sequencing data jointly reveals VDAC2's impacts on prognosis and immune landscape of NSCLC.

Non-small cell lung cancer (NSCLC) is characterized by stronger metastatic ability and worse prognosis. In NSCLC, hypoxia is a major cause of invasion and metastasis through promoting angiogenesis. In present study, NSCLC cell clusters were extracted from single cell-sequencing dataset GSE131907, which were combined with hypoxia-related genes to group clusters. qRT-PCR and western blot were used to validate the expression of target gene. Nine NSCLC clusters were extracted, which were divided into two hypoxia-related subgroups, C1 and C2. Totally 101 differentially expressed prognostic genes were identified between subgroups. Of which, VDAC2 showed excellent prognostic value for NSCLC and was selected for further analysis. VDAC2 was upregulated in tumor samples in TCGA and was correlated with advanced stages. In vitro experiments validated this trend. Five crucial immune cells showed differential infiltration proportions between high and low VDAC2 expression groups. VDAC2 knockdown significantly inhibited the proliferation and invasion ability of NSCLC cells. Integrating single cell and bulk sequencing data as well as wet lab experiments, hypoxia-related VDAC2 exhibited important prognostic value and showed the promise of becoming immune-therapy target in NSCLC.

Fibroblast activating protein promotes the proliferation, migration, and activation of fibroblasts in oral submucous fibrosis.

OBJECTIVES: Fibroblast activating protein (FAP) is associated with various organ fibrosis. However, the expression and molecular function of FAP in oral submucous fibrosis (OSF) is still unclear. MATERIALS AND METHODS: The high-performance liquid chromatography was used to detect the presence of alkaloids in areca nut extract (ANE). Real-time qPCR, Western blot, and Immunohistochemistry assay were used to analyze the expression of FAP mRNA or protein in OSF and normal oral tissue. A chi-squared test analyzed the relationship between FAP protein expression and clinicopathological data of OSF patients. CCK-8, Wound-healing, and Transwell migration assay were employed to assess the effect of the proliferation and migration ability of hOMF cells with FAP overexpression or knockdown. The expression level of a-SMA, FSP1, and P13K-Akt signaling pathways-related protein in hOMF cells transfected with FAP overexpression or knockdown plasmid was verified by western blot assay. RESULTS: The four specific areca alkaloids (Arecoline, Guvacine, Arecaidine, and Guvacoline) were successfully detected in the ANE. The viability of hOMF cells was significantly improved in the 50 µg/mL ANE group and was inhibited in the 5 and 50 mg/mL ANE groups. The expression of FAP was upregulated in OSF tissues, and hOMF cells treated with 50 µg/mL ANE and was related to pathology grade, clinical stage, and history of chewing betel nut. Additionally, FAP may promote the proliferation, migration, and activation of hOMF cells through the P13K-Akt signaling pathway. CONCLUSIONS: This study found that ANE had a bidirectional effect on the viability of hOMF cells, and the FAP gene was a potential therapeutic target in OSF.

Two-dimensional nanomaterials: synthesis and applications in photothermal catalysis.

Photothermal catalysis, as one of the emerging technologies with synergistic effects of photochemistry and thermochemistry, is highly attractive in the fields of environment and energy. Two-dimensional (2D) nanomaterials have received extensive attention toward photothermal catalysis because of their ultrathin layer structures, superior physical and optical properties, and high surface areas. These merits are beneficial for shortening the transfer distance of charge carriers, improving the efficiency of solar to thermal, and providing a great opportunity for the development of photothermal chemistry. In this review, we have summarized the state-of-art advances in various 2D nanomaterials with emphasis on the driving force and relevant mechanism of photothermal catalysis, including the involved three types, namely, localized surface plasmonic resonance (LSPR), nonradiative relaxation, and thermal vibrations of molecules. Moreover, the synthesis strategies of 2D materials and their photothermal applications in carbon dioxide (CO2) conversion, hydrogen (H2) production, volatile organic compounds (VOCs) degradation, and water (H2O) purification have been discussed in detail. Ultimately, the existing challenges and prospects of future development in the field are proposed. It is believed that this review will afford a great reference for the exploration of the high-efficiency 2D nanomaterials and their structure-activity relationship.

Non-linear association between Mediterranean diet and depressive symptom in U.S. adults: A cross-sectional study.

The Mediterranean diet (MED), a dietary pattern rich in fruits and vegetables, whole grains, legumes, nuts, fish, and olive oil, has anti-oxidative and anti-inflammatory effects. Although some data suggest that MED adherence is associated with decreased manifestation of depressive symptoms, it remains necessary to further analyze this apparent non-linear association as well as the influence of different factors on the relationship between MED and depression. Here, we investigated associations between the alternate MED (aMED) score and depressive symptom via multivariate logistic regression, weighted generalized additive (GAM) and two-step linear regression models, analyzing data from the 2005-2018 National Health and Nutrition Examination Survey (NHANES). The most important factor relevant to aMED score that contributed to the prevalence of depressive symptom was assessed using random forest. Furthermore, we examined whether the relationship between aMED score and depressive symptom differs by age, race, sex, socioeconomic variables, lifestyle- and health-related variables, and chronic medical conditions, via subgroup analyses. A total of 19,477 participants (20-80 years of age) were included in this cross-sectional study. In crude and adjusted (1-5) multivariate logistic regression models, increased aMED score was noted to associate with non-depressive status, as defined using the Patient Health Questionnaire-9 (P < 0.05). Data analyses via GAM and two-piecewise linear regression revealed a non-linear association between aMED and depressive symptom, which had an inflection point of 3. Random forest results revealed that vegetable score contributes greatest to the relationship between aMED and depressive symptom. Subgroup analyses revealed that aMED score is significantly negatively related with depressive symptom in most different populations (P < 0.05) with the exception of high annual income, diabetes, borderline blood glucose level and Parkinson's disease (PD) (P > 0.05). In conclusion, we observed a non-linear association between aMED score and depressive symptom. Further studies are needed to validate our results.

Salviae miltiorrhizae and ligustrazine hydrochloride injection combined with mecobalamin for treating diabetic peripheral neuropathy: A protocol for systematic review and meta-analysis.

OBJECTIVE: Currently, it is unclear whether the salviae miltiorrhizae (Danshen Salvia) and ligustrazine hydrochloride (Chuanxiong Chuanxiong) (SMLH) injection combined with mecobalamin can improve diabetic peripheral neuropathy (DPN). We conducted a systematic analysis to evaluate the clinical effects of SMLH injection combined with mecobalamin for treating DPN. METHODS: Seven databases, including PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wan Fang Database (Wang Fang), Chinese Biomedical Literature Database (CBM), and VIP Database for Chinese Technical Periodicals (VIP) were searched for systematic literature retrieval. Each database was searched up to 2020 to identify randomized controlled trials on DPN treated with SMLH injection combined with mecobalamin. We used the RevMan 5.3 and Stata 14.0 software to assess the risk of bias in the included trials. RESULTS: A total of 15 publications, including 1349 samples, were reviewed. The total effective rate of SMLH injection combined with mecobalamin was 31% higher than that of mecobalamin alone (95% confidence interval [CI] = 1.23-1.38; P < .00001). The experimental group showed a significant increase in the motor conduction velocity (MCV) of the peroneal nerve (weighted mean difference [WMD] = 4.81, 95% CI 3.53-6.09; P < .00001). In addition, SMLH injection combined with mecobalamin showed a statistical significant effect on the sensory conduction velocity (SCV) of the peroneal nerve (WMD = 5.03, 95% CI = 4.16-5.90; P < .00001), and MCV of the median nerve (WMD = 5.38, 95% CI = 4.05-6.72; P < .00001). The WMD for the change in SCV in the median nerve was 4.89 m/s (95% CI = 3.88-5.89; P < .00001). The P-values of the Egger and Begg tests were 0.967 and 0.961, respectively, indicating no publication bias. Subgroup and sensitivity analyses indicated that the results for MCV and SCV of the peroneal nerve and the median nerve were stable. CONCLUSION: SMLH injection combined with mecobalamin can improve DPN, compared with mecobalamin alone.

[Meta-analysis of effect of Jinqi Jiangtang Tablets on treating insulin resistance in type 2 diabetes].

To systematically assess the efficacy and safety of Jinqi Jiangtang Tablets for patients with insulin resistance in type 2 diabetes, literatures were retrieved in 7 databases: PubMed, EMbase, Cochrane Library, Chinese National Knowledge Infrastructure(CNKI), WanFang database, Chinese BioMedical Database(CBM), VIP Chinese Science and Technique Journals Database, from the date of its inception up to November 2018. Review Manager 5.3 software was used for risk bias assessment, data synthesis and subgroup analysis. Begg's and Egger's tests were performed for assessing symmetries of funnel plot by software Stata 14.0. GRADE system was used to assess the quality of evidence. A total of 10 trials involving 797 participants were eligible. Compared with Western medicine alone, Jinqi Jiangtang Tablets showed a statistical significance in FBG(WMD=-0.63, 95%CI[-1.00,-0.26]). Jinqi Jiangtang Tablets showed a significant decrease in 2 h BG combined with Western medicine compared with Western medicine alone(WMD=-1.46, 95%CI[-1.71,-1.21], P<0.000 01). Jinqi Jiangtang Tablets combined with Western medicine showed a significant decrease in HbA1 c(WMD=-0.75, 95%CI[-0.97,-0.53], P<0.000 01), FINS(WMD=-0.65, 95%CI[-0.80,-0.50], P<0.000 01), 2 h INS(SMD=-1.67, 95%CI[-2.26,-1.09], P<0.000 01) and HOMA-IR(WMD=-1.22, 95%CI[-1.67,-0.76], P<0.000 01). Jinqi Jiangtang Tablets combined with Western medicine was beneficial for ISI(WMD=1.00, 95%CI[0.84, 1.17], P<0.000 01). Egger's and Begg's test showed no publication bias(P=0.379). Sensitivity analysis showed no impact on the overall results. The GRADE quality of the evidence was low. Despite of the apparently positive results, we cannot draw a rational conclusion that Jinqi Jiangtang Tablets has a positive effect in patients with IR, because of the low evidence grade.

The repertoire of tumor-infiltrating lymphocytes within the microenvironment of oral squamous cell carcinoma reveals immune dysfunction.

BACKGROUND: The role of tumor-infiltrating lymphocytes (TILs) in the immune remodeling of tumor microenvironments (TME) in oral squamous cell carcinoma (OSCC) remains controversial. In this study, we pursued a comprehensive characterization of the repertoire of TILs and then analyzed its clinical significance and potential prognostic value. METHODS: Fresh tumor tissue samples and peripheral blood from 83 OSCC patients were collected to comprehensively characterize the phenotypes and frequencies of TILs by flow cytometry. Archived paraffin-embedded tissues derived from 159 OSCC patients were analyzed by immunohistochemistry to further assess the TIL repertoire. The clinical significance of TILs and their potential prognostic value were further analyzed. RESULTS: A series of unique features of TILs were observed. IL-17 was highly expressed in betel nut chewers, and CD20 was abundantly expressed in patients who did not drink alcohol; high expression of CD138, PD-L1, and Foxp3 was associated with poor prognosis. The Th17/Treg ratio was an independent prognostic factor for patient survival with greater predictive accuracy for overall survival. CONCLUSIONS: Our results suggest an antigen-driven immune response; however, the immune dysfunction within the microenvironment in OSCC and the Th17/Treg balance may play important roles in the modulation of antitumor immunity.

A systematic review of randomized controlled trials of the Wenyang Huoxue method in treating diabetic peripheral neuropathy.

OBJECTIVE: To assess the efficacy and safety of the Wenyang Huoxue method for patients with diabetic peripheral neuropathy. METHODS: A systematic literature search was performed using 7 databases: PUBMED, EMBASE, the Chinese National Knowledge Infrastructure, Wanfang, Chinese BioMedical, and the VIP Chinese Science and Technique Journals. The publication time was from the start of each database up to November 2018. Review Manager 5.3 software was used for assessing potential bias, data synthesis, and the subgroup analysis. Begg and Egger tests were used to assess funnel plot symmetries using Stata 14.0 software. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) system was used to assess the quality of evidence. RESULTS: A total of 22 trials involving 1835 participants were eligible. There were significant differences in a total effective rate between the Wenyang Huoxue method combined with Western medicine and Western medicine alone (RR = 1.33, 95% CI 1.26-1.41; P < .00001). As for the sensory conduction velocity (SCV) of the peroneal nerve, the Wenyang Huoxue method combined with Western medicine compared with Western medicine alone had a significant increase (weighted mean difference [WMD] = 5.00, 95% CI 3.42-6.57; P < .00001). Also, the Wenyang Huoxue method combined with Western medicine had significant increases in motor conduction velocity (MCV) of the peroneal nerve (WMD = 4.48, 95% CI 3.78-5.19; P < .00001), tibial nerve SCV (WMD = 3.47, 95% CI 2.66-4.28; P < .00001), tibial nerve MCV (4.87, 95% CI 3.21-6.53; P < .00001), median nerve SCV (WMD = 3.78, 95% CI 3.07-4.50; P < .00001), and median nerve MCV (WMD = 4.50, 95% CI 3.40-5.59; P < .00001). However, the effect of the Wenyang Huoxue method on fasting blood glucose, 2-h postprandial blood glucose, and glycosylated hemoglobin was not statistically significant. Egger's test results showed that there was no publication bias (P = .0008), but the trim and filling method showed steady results. An influence analysis showed that no single study affected the overall result. The GRADE quality of the evidence was low to moderate across the different outcomes. CONCLUSION: Despite of the apparently positive findings, the quality of GRADE is not high, suggesting that the Wenyang Huoxue method can improve nerve conduction velocity to a certain extent, but more rigorous literature is needed to support this evidence.

Xueshuantong for Injection Ameliorates Diabetic Nephropathy in a Rat Model of Streptozotocin-Induced Diabetes.

Diabetic nephropathy (DN) is a major complication of diabetes and becomes the chief cause of end-stage renal disease. Our study was undertaken to investigate the ameliorative effect and underlying mechanism of Xueshuantong for Injection (XST) on DN in streptozotocin (STZ)-induced rats. Effect of XST treatment (XST, 50 mg/kg/day, i.p.) lasting 60 days after STZ-induced (60 mg/kg, i.p.) diabetes was investigated. Blood sugar levels and body weight were recorded every week of the experiment. At the 28th and 56th days after injection urine glucose and 24 h urine protein excretion were determined. Apoptosis related factors such as cleaved caspase-3, Bcl-2, Bax and inflammation related factors, including tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), IL-1ß, inducible nitric oxide synthase (iNOS) and intercellular adhesion molecule-1 (ICAM-1) were detected by PCR or western blot. The expression levels of fibronectin, Collagen Ⅰ, α-smooth muscle actin (α-SMA) and proliferating cell nuclear antigen (PCNA), TGF-ß-Smad2/3 signaling pathway, and receptor for advanced glycation end products (RAGE) was investigated. Our results showed that XST treatment did not affect levels of body weight, blood glucose and urine glucose levels. Our analysis revealed that XST inhibited cell apoptosis and suppressed the properties of RAGE in the kidney. XST treatment could also significantly suppress the overexpression of pro-inflammatory mediators in kidney and prevent renal fibrosis by blocking the TGF-ß/Smad2/3 signaling pathway. In conclusion, our findings suggested that XST could provide protection against DN through reduction of RAGE accumulation, decreasing inflammation, inhibition of renal fibrosis, and blocking the TGF-ß/Smad2/3 signaling pathway.

Mitogen-activated protein kinase signaling pathway in oral cancer.

The mitogen-activated protein kinase (MAPK) signaling pathway is associated with tumor cell proliferation, differentiation, apoptosis, angiogenesis, invasion and metastasis. The present review assesses the involvement of the MAPK signaling pathway in oral cancer progression and invasion based on analysis of individual sub-pathways and their mechanisms of action. The regulation of this pathway for targeted oral cancer therapy is explored and the challenges confronting this, as well as corresponding potential solutions, are discussed. Exploring this pathway with an emphasis on its components, subfamilies, sub-pathways, interactions with other pathways and clinical practice modes may improve oral cancer treatment.

[Anterolateral thigh flap for reconstruction of defects after en bloc resection of buccal cancer : a retrospective study of 23 cases].

PURPOSE: This study was aimed to observe the effects of anterolateral thigh flap for reconstruction of tissue defects after en bloc resection of buccal cancer. METHODS: Twenty-three patients with soft tissue defects after en bloc resection of buccal cancer underwent simultaneous reconstruction with anterolateral thigh flap from May, 2013 to May, 2015 were observed. Anterolateral thigh flaps were designed and harvested in form of single or multiple islands to restore the defect in buccal region after surgery. The appearance and function of both the oral and maxillofacial region and the donor site were recorded and evaluated. RESULTS: All the 23 flaps survived. Only 3 of them experienced vascular crisis within 24 hours after surgery, and recovered gradually after salvage. The success rate was 100%. One to three years of follow-up showed satisfying morphology and function for both the receipt sites and the donor sites. Buccal abscess was observed in 1 patient and recovered after rinsing and drainage. Two patients died of recurrence. CONCLUSIONS: Good effects can be achieved using anterolateral thigh flap to reconstruct buccal defects after en bloc resection of cancer, which is suitable for application in clinical practice.

Circadian clock components RORα and Bmal1 mediate the anti-proliferative effect of MLN4924 in osteosarcoma cells.

The anticancer small molecule MLN4924, a Nedd8-activating enzyme (NAE) inhibitor, triggers cell-cycle arrest, apoptosis, and senescence in cancer cells. In this study, we demonstrate that MLN4924 suppresses osteosarcoma cell proliferation by inducing G2/M cell cycle arrest and apoptosis. Our results indicate that MLN4924 stabilizes the retinoid orphan nuclear receptor alpha (RORα) by decreasing its ubiquitination. RNA interference of RORα attenuates the anti-proliferative effect of MLN4924 in U2OS osteosarcoma cells. MLN4924 up-regulates the expression of p21 and Bmal1, two transcriptional targets of RORα. However, p21 plays a minimal role in the anti-proliferative effect of MLN4924 in U2OS osteosarcoma cells. In contrast, Bmal1 suppression by siRNA attenuates the anti-proliferative effect of MLN4924 in U2OS osteosarcoma cells, indicating that the MLN4924-mediated cell growth inhibition is mediated by Bmal1. These results show MLN4924 to be a promising therapeutic agent for the treatment of osteosarcoma and suggest that MLN4924-induced tumor growth inhibition is mediated by the circadian clock components RORα and Bmal1.

Investigation of the association between miR­181b, Bcl­2 and LRIG1 in oral verrucous carcinoma.

Abnormal expression of microRNAs (miRNAs) is involved in the development of and anti­apoptotic effects in various types of human cancer. However, miRNA­mediated regulation of oral verrucous carcinoma (OVC) remains to be elucidated. The present study aimed to investigate the expression of miR­181b in OVC and oral squamous cell carcinoma (OSCC). The expression levels of miR­181b were determined using reverse transcription­quantitative polymerase chain reaction. The expression levels of B­cell lymphoma 2 (Bcl­2) and leucine rich repeats and immunoglobulin like domains 1 (LRIG1), were evaluated using immunohistochemical staining. The correlation between Bcl­2 and LRIG1 expression was determined using a Pearson correlation analysis. The expression levels of miR­181b and Bcl­2 in OVC were significantly higher compared with normal mucosal tissue (NM); however, lower compared with the OSCC. The key target of miR­181b was LRIG1 and it was significantly lower in OVC tissues compared with NM tissue; however this was higher when compared with OSCC tissue. The expression levels of Bcl­2 were correlated with expression levels of LRIG1 in OVC tissues. Therefore, LRIG1 may be associated with anti­apoptotic function in OVC tissues.

An adaptive immune response driven by mature, antigen-experienced T and B cells within the microenvironment of oral squamous cell carcinoma.

Lymphocyte infiltrates have been observed in the microenvironment of oral cancer; however, little is known about whether the immune response of the lymphocyte infiltrate affects tumor biology. For a deeper understanding of the role of the infiltrating-lymphocytes in oral squamous cell carcinoma (OSCC), we characterized the lymphocyte infiltrate repertoires and defined their features. Immunohistochemistry revealed considerable T and B cell infiltrates and lymphoid follicles with germinal center-like structures within the tumor microenvironment. Flow cytometry demonstrated that populations of antigen-experienced CD4+ and CD8+ cells were present, as well as an enrichment of regulatory T cells; and T cells expressing programmed death-1 (PD-1) and T cell Ig and mucin protein-3 (Tim-3), indicative of exhaustion, within the tumor microenvironment. Characterization of tumor-infiltrating B cells revealed clear evidence of antigen exposure, in that the cardinal features of an antigen-driven B cell response were present, including somatic mutation, clonal expansion, intraclonal variation and isotype switching. Collectively, our results point to an adaptive immune response occurring within the OSCC microenvironment, which may be sustained by the expression of specific antigens in the tumor.