TRK-fused gene (TFG) regulates ULK1 stability via TRAF3-mediated ubiquitination and protects macrophages from LPS-induced pyroptosis.

TRK-fused gene (TFG) is known to be involved in protein secretion and plays essential roles in an antiviral innate immune response. However, its function in LPS-induced inflammation and pyroptotic cell death is still unknown. Here, we reported that TFG promotes the stabilization of Unc-51 like autophagy activating kinase (ULK1) and participates in LPS plus nigericin (Ng) induced pyroptotic cell death. Our results showed that TFG-deficient THP-1 macrophages exhibit higher mitochondrial ROS production. LPS/Ng stimulation triggers a much higher level of ROS and induces pyroptotic cell death. ULK1 undergoes a rapid turnover in TFG-deficient THP-1 cells. TFG forms complex with an E3 ligase, tumor necrosis factor receptor-associated factor 3 (TRAF3), and stabilizes ULK1 via disturbing ULK1-TRAF3 interaction. Knockdown of TFG facilitates the interaction of ULK1 with TRAF3 and subsequent K48-linked ULK1 ubiquitination and proteasome degradation. Rescue of ULK1 expression blocks LPS/Ng-induced cell death in TFG-deficient THP-1 macrophages. Taken together, TFG plays an essential role in LPS/Ng-induced pyroptotic cell death via regulating K48-linked ULK1 ubiquitination in macrophages.

Helmet chinstrap protective role in maxillofacial blast injury.

BACKGROUND: The protective role of helmet accessories in moderating stress load generated by explosion shock waves of explosive devices is usually neglected. OBJECTIVE: In the presented study, the protective role of the helmet chinstrap against the impulse and overpressure experienced by the maxillofacial region were examined. METHODS: The explosion shock wave and skull interaction were investigated under three different configurations: (1) unprotected skull, (2) skull with helmet (3) skull with helmet and chinstrap. For this purpose, a 3D finite element model (FEM) was constructed to mimic the investigated biomechanics module. Three working conditions were set according to different explosive charges and distances to represent different load conditions. Case 1: 500 mg explosive trinitrotoluene (TNT), 3 cm, case 2: 1000 mg TNT, 3 cm, and case 3: 1000 mg TNT and 6 cm distance to the studied object. The explosion effect was discussed by examining the shock wave stress flow pattern. Three points were selected on the skull and the stress curve of each point position were illustrated for each case study. RESULTS: The results showed that the helmet chinstrap can reduce the explosive injuries and plays a protective role in the maxillofacial region, especially for the mandible.

Associations between osteoporosis and risk of periodontitis: A pooled analysis of observational studies.

BACKGROUND: Periodontitis and osteoporosis are most popular among aging population and both conditions might be linked, even though, this suggestion still until now debated. OBJECTIVES: A meta-analysis on previous investigations has been used to evaluate the correlation between periodontitis and osteoporosis to determine whether osteoporosis is a local indicator of bone loss, or whether it is depending on or related to periodontitis causes. METHODS: The literature database, including but not excluding, The Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, CINAHL, and Science Citation Index Expanded, was searched in this work during Feb, 2020. We conducted the investigations contain cohort studies, cross-sectional studies, as well as case-control studies with relative risk (RR) or odds ratio (OR) and 95% confidence intervals (CIs). Subgroup and Sensitivity analysis were also applied to identify heterogeneity sources. RESULTS: 23 observational studies with 12 cohorts, 7 cross-sectional and 4 case-control studies, were included, together with 2,157,037 participants. Osteoporosis patients were more exposed to periodontitis (OR, 1.96; 95% CI, 1.50-2.54). Subgroup analyses showed that the higher risk of osteoporosis in periodontitis patients exists in both cross-sectional studies (OR, 2.17; 95% CI, 1.80-2.61) and case-control studies (OR 2.63; 95% CI, 1.69-4.09), and marginally in cohort studies (OR, 1.70; 95% CI, 1.16-2.49). CONCLUSION: Review analyses have shown that osteoporosis is closely related to the increased risk of periodontitis in the future. Dental specialists better to understand the potential association between periodontitis and osteoporosis.

Factors Correlated With Internet Gaming Disorder Among Malaysian University Students

@#Introduction: Internet gaming disorder (IGD) has recently been incorporated into the Diagnostic and Statistical Manual of Mental Disorders fifth edition (DSM-5) as a disorder for future research. The primary objectives of the present study are to describe the level of IGD and to examine its correlations with sociodemographic factors and psychological comorbidities among undergraduate students in a Malaysian university. Methods: A total of 411 undergraduate students completed an online questionnaire. They were selected from a random sample of participating university faculties. The online questionnaire contained the Internet Gaming Disorder Scale-Short-Form and the Depression, Anxiety, Stress Scale. Results: The presents study reported that 52.8% of the participants had high IGH. Using hierarchical multiple regression, age (β = -0.09, p < 0.05), gender (β = -0.40, p < 0.001) and stress (β = 0.23, p < 0.05) were statistically significant predictors of IGD. Academic performance, depression and anxiety did not emerge as significant predictors. Conclusion: These findings highlight the risk factors (in particular, stress) of IGD. Further studies on interventions, particular that of preventative strategies, will be needed to combat this emerging public health problem.

Associations Between Poor Oral Health and Risk of Squamous Cell Carcinoma of the Head and Neck: A Meta-Analysis of Observational Studies.

PURPOSE: Many epidemiologic studies have reported an association of poor oral health, especially periodontal disease (PD) and tooth loss, with the risk of squamous cell carcinoma of the head and neck (SCCHN). However, these studies have yielded inconsistent results. Therefore, the present study investigated whether poor oral health is an independent predictor of SCCHN through a meta-analysis of observational studies. MATERIALS AND METHODS: The PubMed, EMBASE, and Cochrane Library databases were systematically searched for relevant observational studies of the association between oral health and risk of SCCHN conducted up to October 2017. The meta-analysis was conducted using STATA 12.0 (StataCorp, College Station, TX). A fixed- or random-effects model was applied to evaluate pooled risk estimates, and sensitivity and subgroup analyses were performed to identify sources of heterogeneity and pooled estimation. Publication bias was assessed using the Begg test, the Egger test, and funnel plots. RESULTS: Twenty-seven relevant observational studies were identified, consisting of 24 case-and-control studies, 2 prospective studies, and 1 cross-sectional study, with 26,750 participants. Notably, oral health correlated meaningfully with SCCHN (odds ratio [OR] = 2.24; 95% confidence interval [CI], 1.77-2.82). In subgroup analyses, participants with PD (OR = 2.52; 95% CI, 1.43-4.44) had a higher risk of developing SCCHN than those with tooth loss (OR = 2.13; 95% CI, 1.63-2.78). The risk estimates exhibited substantial heterogeneity. Evidence of publication bias was limited. CONCLUSIONS: The results of this meta-analysis suggest that patients with tooth loss or PD might face a substantial and independent risk of SCCHN, even after adjusting for smoking and alcohol consumption. However, the pooled estimates from observational studies could not establish a causative relation among PD, tooth loss, and SCCHN. Additional investigations of this correlation are warranted.

[Effect of bone morphogenetic protein-4 overexpression on the biological activity of mouse induced pluripotent stem cells].

OBJECTIVE: This study aimed to construct the expression of bone morphogenetic protein-4 (BMP4) lentiviral vector gene and explore its influence on the biological activity of mouse induced pluripotent stem (iPS) cells. METHODS: iPS cell lines stably overexpressing BMP4 were constructed by lentivirus transfection (BMP4-overexpressing group). Cells without transfection served as the blank group, and cells with only vector transfection served as the empty-vector group. Cell proliferation was detected by CCK8, and the expression levels of ameloblastin (AMBN), cytokeratin (CK) 14, dentin sialophospho-protein (DSPP), bone sialoprotein (BSP), and Runx2 mRNA were detected by quantitative polymerase chain reaction. Alkaline phosphatase (ALP) activity was used to detect the degree of cell differentiation. RESULTS: Compared with blank and empty-vector groups, proliferation activity and ALP activity of BMP4-overexpressing group obvious increased (P<0.05), BMP4, AMBN, CK14, DSPP, BSP, Runx2 mRNA expression also increased (P<0.05). CONCLUSIONS: BMP4 can significantly promote the odontogenic differentiation of iPS.

How Is Third Molar Status Associated With the Occurrence of Mandibular Angle and Condyle Fractures?

PURPOSE: Third molars (M3s) have been hypothesized to be associated with the risk of mandibular angle fracture and mandibular condylar fracture. The authors systematically estimated the relative risk (RR) of M3 status for the development of mandibular angle fracture and mandibular condylar fracture through a meta-analysis of cohort studies. MATERIALS AND METHODS: In this systematic review, the PubMed, EMBASE, and Cochrane Library databases were searched from inception to October 2016. The predictor of risk was the presence or absence of M3s. The primary outcome was the RR of mandibular angle or condylar fracture. A fixed- or a random-effects model was applied to evaluate the pooled risk estimates. Sensitivity analysis also was performed to identify the potential sources of heterogeneity. Publication bias was assessed by the Begg and Egger tests. RESULTS: Overall, 13 retrospective cohort studies were included. Of these, 13 reported the association between M3s and mandibular angle fracture, and 5 reported the association with mandibular condylar fracture. Patients with M3s had an increased risk of mandibular angle fractures (RR = 2.63; 95% confidence interval [CI], 2.15-3.21) but a decreased risk of mandibular condylar fractures (RR = 0.47; 95% CI, 0.25-0.86). Substantial heterogeneity in the risk estimates was found. No evidence of publication bias was found. CONCLUSION: The present meta-analysis provides further evidence associating the presence of M3s with an increased risk of mandibular angle fractures and a simultaneously decreased risk of mandibular condylar fracture. Because of potentially more serious complications associated with condylar fracture, clinicians should carefully consider the decision to remove M3s to decrease the risk of mandibular angle fracture.

Calcitonin gene-related peptide stimulates BMP-2 expression and the differentiation of human osteoblast-like cells in vitro.

AIM: To investigate whether bone morphogenic protein-2 (BMP-2) expression was involved in calcitonin gene-related peptide (CGRP)-induced osteogenesis in human osteoblast-like cells in vitro. METHODS: MG-63 osteogenic human osteosarcoma cells were treated with CGRP (10-8 mol/L) for 48 h. Cell cycle phases were determined using flow cytometry assay. The protein levels of BMP-2, ALP, Osteocalcin, ColIa1, CREB, and pCREB were measured with Western blotting, while the mRNA level of BMP-2 was measured with qR-T PCR. The expression of ALP in MG-63 cells was also studied using immunofluorescence staining. The level of cAMP was measured with ELISA assay. RESULTS: CGRP treatment significantly stimulated proliferation of MG-63 cells, and increased the expression of BMP-2 and the osteogenic proteins ALP, Osteocalcin and ColIa1. Pretreatment with the BMP signaling inhibitor Noggin (100 ng/mL) did not affect CGRP-stimulated proliferation and BMP-2 expression, but abolished the CGRP-induced increases of the osteogenic proteins ALP, Osteocalcin and ColIa1. Furthermore, CGRP treatment markedly increased cAMP level in MG-63 cells, whereas pretreatment with the cAMP pathway inhibitor H89 (5 µmol/L) abolished the CGRP-induced increases of cAMP level and BMP-2 expression. CONCLUSION: In MG-63 cells, the BMP pathway is involved in CGRP-induced osteogenic differentiation but not in proliferation, whereas the cAMP/pCREB pathway is involved in the expression of BMP-2.

20-Hydroxyecdysone-induced bone morphogenetic protein-2-dependent osteogenic differentiation through the ERK pathway in human periodontal ligament stem cells.

20-Hydroxyecdysone, an ecdysteroid hormone, can induce osteogenic differentiation in mesenchymal stem cells. Periodontal ligament stem cells (PDLS cells) have mesenchymal-stem-cell-like qualities and are considered as one of the candidates of future clinical application in periodontitis treatment. However, there are no studies describing the effect of 20-Hydroxyecdysone on PDLS cells. In this paper, we report a detailed study on the effect of 20-Hydroxyecdysone on PDLS cell proliferation in vitro. PDLS cells were developed from human PDL cells and were treated with 20-Hydroxyecdysone to understand different aspects of its effects. 20-Hydroxyecdysone promoted PDLS cell proliferation; significantly increased the gene expression levels of runt-related transcription factor 2, alkaline phosphatase (ALP), type I collagen, and osteocalcin. Moreover, 20-Hydroxyecdysone enhanced bone formation by PDLS cells and significantly increased bone morphogenetic protein-2 (BMP-2) mRNA and protein expression. However, 20-Hydroxyecdysonemediated increase in ALP activity was blocked with a BMP-2-specific neutralizing antibody or with the antagonist noggin; and20-Hydroxyecdysone mediated induction of BMP-2 expression and increase of ALP activity were abolished by the extracellular regulated protein kinase (ERK) MAPK pathway inhibitor PD98059. 20-Hydroxyecdysone also increased the phosphorylation of ERK. These findings provide evidence to state that 20-Hydroxyecdysone stimulates cell proliferation and induces osteogenic differentiation through the induction of BMP-2 expression in PDLS cells. It also shows that the ERK pathway is involved in 20-Hydroxyecdysone induced BMP-2 expression and osteogenic differentiation. These results are suggesting its potential as a drug for periodontal regenerative therapy.

Lack of association of the cyclin D1 G870A variation with oral carcinoma risk: Evidence from 2,404 subjects.

Evidence implicates cyclin D1 (CCND1) G870A polymorphisms as risk factors for various cancers. An increasing number of investigations have been conducted on the association of CCND1 G870A polymorphisms with susceptibility to oral carcinoma, and have yielded inconclusive results. The aim of the present study was to derive a more precise estimation of the correlation. Meta-analyses examining the association between CCND1 G870A polymorphisms and oral carcinoma were performed. Separate analyses on ethnicity, smoking status and control sources were also implemented. Eligible studies were identified prior to February 2012. From the overall data from 1,128 cases and 1,276 controls, no associations of CCND1 G870A polymorphisms with oral carcinoma were observed [AA vs. GG: odds ratio (OR)=1.06; 95% confidence interval (CI), 0.62-1.82; dominant model: OR=1.04; 95% CI, 0.76-1.43; recessive model: OR=1.06; 95% CI, 0.70-1.59]. In the subgroup analysis by ethnicity, smoking status and control sources, no significant associations of CCND1 G870A polymorphisms and oral cancer were observed for the three genetic models. Collectively, the data failed to suggest CCND1 G870A polymorphism as a low-penetrant risk factor for developing oral carcinoma. Additional studies with large sample sizes concerning different ethnicities in different areas are required.

[Influence of receptor activity modifying protein 1 overexpression on enhancing effect of calcitonin gene-related peptide on MG-63 cells proliferation].

OBJECTIVE: To investigate the influnce of receptor activity modifying protein 1(RAMP-1) overexpression on enhancing effect of calcitonin gene-related peptide on MG-63 cells proliferation. METHODS: Cultured MG-63 osteoblasts in exponential phase of growth were randomly divided into three groups: RAMP-1 overexpression group, empty vector control group and negative control group. RAMP-1 eukaryotic expression vector was constructed and stably transfected into MG-63 cells. Realtime-polymerase chain reaction, Western blotting and immunofluroescence were used respectively to detect the expression of calcitonin receptor-like receptor (CRLR) in the cells and its distribution on cell membrane. The status of proliferation was detected respectively at 0, 24, 48, 72, 96 h by cell counting kit-8 (CCK-8) and cells were collected to analyze their cycle respectively at 0, 8, 16, 24 h by flow cytometry. RESULTS: CRLR protein and mRNA expression levels of MG-63 cells in RAMP-1 overexpression group were significantly higher than the other two groups (P < 0.05). The A value of RAMP-1 overexpression group at 24, 48, 72, 96 h were 0.628 ± 0.175, 0.896 ± 0.592, 1.055 ± 0.004, 1.179 ± 0.618, respectively, which were significantly higher than that of the other two groups (P < 0.05). The difference was most pronounced at 72 h. S-phase fraction of RAMP-1 overexpression group was (1.25 ± 0.13)%, (68.79 ± 0.56)%, (64.49 ± 1.59)%, (57.82 ± 0.75)%, respectively, which were significantly higher than the other two groups (P < 0.05). The difference was most pronounced at 8 h. CONCLUSIONS: RAMP-1 overexpression can promote CRLR distribution on MG-63 cell membrane and enhance CGRP's promotion effect on MG-63 cell proliferation.

NAT2 polymorphisms with oral carcinoma susceptibility: a meta-analysis.

Published data have implicated NAT2 polymorphisms as risk factors for various cancers. A number of studies have focused on the association of NAT2 polymorphisms with susceptibility to oral carcinoma and have yielded inconclusive results. The aim of the present study was to derive a more precise estimation of the relationship. We first carried out a deliberate search in the databases without a language limitation, covering all papers published up to Dec 2011. A total of seven case-control studies including 1,379 cases and 1,868 controls were selected and the relevant data were extracted for systematic meta-analyses. No significant association was found for the overall data (OR: 1.04, 95 % CI: 0.79-1.39). In subgroup analyses according to ethnicity, slow acetylators might increase oral cancer risk among Asians (OR: 1.38, 95 % CI: 1.04-1.82) but not Caucasians or Mixed races. The data suggested that NAT2 polymorphisms might be a low-penetrant risk factor for oral carcinoma in Asians.

[Computer simulation of projectile injuries to pig mandibular angle].

OBJECTIVE: To investigate the feasibility of computer simulation in maxillofacial firearm injury. METHODS: The three-dimensional finite element models and simulations of 7.62 mm, 5.56 mm standard bullets projectile injuries to pig mandibular angle were established by using MIMICS, ANSA, LS-DYNA and LS-POST software. Based on the simulation results, the bullet hole diameters, energy loss values, energy loss rates, von Mises stress, effective strain, effective strain rate dynamic contours at different time points were used for biomechanical analysis. RESULTS: The damage processe of 7.62 mm, 5.56 mm standard bullets projectile injury to pig mandibular angle were simulated successfully. The injury rate of 7.62 mm standard bullet and injury severity of the mandible were higher than that of 5.56 mm standard bullet. CONCLUSIONS: Computer simulation can simulate maxillofacial firearm injuries effectively and may become an important method for oral and maxillofacial firearm injuries analysis.

[Effect of Notch ligand Delta1-RNA interference by lentivirus on proliferation and differentiation of human dental pulp stem cells].

OBJECTIVE: To investigate the effects of specific RNA interference (RNAi) to Notch ligand Delta1 on proliferation and differentiation of human dental pulp stem cells (DPSC). METHODS: DPSC were infected by lentivirus vectors carrying Delta1-RNAi. DPSC were divided into three groups, DPSC/Delta1-RNAi group, DPSC/wt group and DPSC/vector group as control. Cell counting kit-8 (CCK-8) assay and flow cytometry were used to evaluate the proliferation of DPSC. Expression of proliferating cell nuclear antigen (PCNA) was examined with immunohistochemical staining. All groups were cultured in an odonto-inductive medium and were observed under microscope. The number of mineralization nodules was counted after Alizarin red staining. Alkaline phosphatase (ALP) activity and the expression of dentin sialophosphoprotein (DSPP) were detected by ALP activity assay and Western blotting. RESULTS: Compared with DPSC/wt or DPSC/vector separately, proliferating rate and S-cycle of DPSC/Delta1-RNAi was significantly lower. The S phase and proliferation index (PI) decreased markedly from 22.32 ± 2.35 and 33.68 ± 4.19 (DPSC/Delta1-RNAi) to 5.44 ± 0.91 and 16.00 ± 6.07 (DPSC/wt). The PCNA staining of DPSC/Delta1-RNAi was evidently weaker. DPSC/Delta1-RNAi group had more calcified cell nodules than the other two control groups, and ALP activity and DSPP expression of DPSC/Delta1-RNAi group increased markedly. CONCLUSIONS: Delta1-RNAi induced by the lentivirus vectors may inhibit DPSC proliferation and differentiation. Notch-Delta signal pathway plays an important role in self-renewal and differentiation.

[Isolation and identification of huamn dental pulp stem cells].

OBJECTIVE: To isolate and identify human dental pulp stem cells from third molars. METHODS: Dental pulps were dissected and digested by collagenase type I and dispase. The obtained single cell supernatant were harvested and cultured. Characterization of the phenotype of DPSCs was detected by immunohistochemical method and RT-PCR assay. Cell cycle was analyzed by FCM. Differentiation potential of DPSCs was evaluated. RESULTS: Colony-forming efficiency of cells derived from dental pulp tissue was 2 - 15 clones/10(3) cells plated. DPSCs were found to express many different markers, including vimentin, collagen type I, GFAP, nestin and osteocalcin, while they failed to react with MyoD and DSPP. About 64.1% of the cells were in G0/G1 phases, while only 35.8% in proliferation (S + G2 + M). Grown in an adipogenic cocktail medium for three weeks, some DPSCs expressed fat cell markers of PPARgamma and LPL, and formed oil red O-positive lipid clusters in five weeks. After culture with a myogenic-inductive medium, DPSCs were found to express MyoD, desmin and myosin, markers of myocyte. Long-term cultures of DPSCs grown in differentiation inductive medium demonstrated the capacity to form Von Kossa-positive condensed nodules with high levels of calcium. CONCLUSION: Cells isolated from adult human dental pulp are clonogenic, and have multipotent differentiation potential, satisfying the criteria of postnatal somatic stem cell.