Objective: To investigate the regulatory effect of miRNA on chronic masticatory myalgia(CMM). Methods: Twenty-three CMM patients were recruited from the Center for TMD and Orofacial Pain of Peking University School and Hospital of Stomatology, 23 gender- and age- matched healthy people recruited as control group. A total of 2 mL peripheral blood was collected from each participant of two groups. MiR-15a-5p and miR-19b-3p were preliminarily screened out as target miRNAs which are associated with CMM by regulating MEK3 according to Targetscan database and DIANA database in the previous experiments. QRT-PCR was used to select the possible miRNA further. Subsequently, the cell transfection and miRNA-overexpression experiment were conducted to verify the relationship of miRNA and MEK3 gene with qRT-PCR and Western blot. Results: MiR-19b-3p expressions were significantly decreased in CMM group compared to healthy control group using qRT-PCR assays (P<0.05), and miR-15a-5p expressions were not significantly different between these two groups(P>0.05). MiR-19b-3p was selected to verify the relationship between miRNA and MEK3 gene in the following experiments. The results showed that the mRNA and protein of MEK3 gene had no significant difference between miR-19b-3p-overexpression group and miR-NC group in human leukemia monocytic cell line (THP-1 cell) at 24h、48h and 72h ( P>0.05). Conclusions: MiR-19b-3p may be involved in CMM and its mechanism is likely to be complicated. It may not work by binding to the MEK3 gene directly.

Objective: We collected a family with non-syndromic oligodontia, and tried to detect the genetic etiology. Methods: After the medical history, clinical examinations including imaging examination, and blood sample were collected, we isolated the genomic DNA and conducted the whole exome sequence (WES). All the results were aligned to the normal human genome, and verified by Sanger sequencing. A series of bioinformatics database such as Exome Aggregation Consortium (ExAC), Genome Aggregation Database (gnomeAD), Single Nucleotide Polymorphism Database (dbSNP), Sorting Intolerant from Tolerant (SIFT), polymorphism phenotyping (PolyPhen-2), and Mutation Taster, were used for mutation screening and confirmation. Results: The proband of the family was congenital missing 22 permanent teeth, and his parents and little brother showed normal clinical manifestations. The consequences of WES indicated that the proband carried compound heterozygous WNT10A mutations [c.637A>G (p.G213S) and c.985C>T (p.R329X)]. Further studies confirmed that the proband's missense mutation c.637A>G(p.G213S) was inherited from his father, while the nonsense mutation c.985C>T (p.R329X) was inherited from his mother. The little brother did not carry any mutations. Conclusions: Our study revealed a novel WNT10A compound heterozygous mutation and its genetic origin in a family with non-syndromic oligodontia. This result can be used for genetic counseling and prenatal diagnosis, and will improve understanding the role of WNT10A mutations in the pathogenesis of congenital tooth agenesis.

Objective: The goal of this study was to identify the potential curcumin binding proteins in oral cancer cells by biotin-streptavidin pull down technique. Methods: A curcumin-biotin conjugate was chemically synthesized and its activity was measured by MTT assay. Curcumin-binding proteins were eluted with streptavidin resin from curcumin-biotin treated oral cancer cells, separated by polyacrylamide gel electrophoresis, and identified by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Curcumin-binding properties of two of these proteins, Annexin A2 and CDKN2B, were studied further by streptavidin binding assay and western blotting with recombinant humum proteins. Results: Curcumin-biotin conjugate was successfully chemically synthesized and its activity was not affected by MTT assay. The potential curcumin binding proteins were eluted with streptavidin resin and separated by SDS-PAGE electrophoresis, and then analyzed by LC-MS/MS. 167 potential curcumin binding proteins were identifed with MASCOT. Binding assay with western blotting confirmed the binding of Annexin A2 and CDKN2B with curcumin in a dose-dependent manner. Conclusions: In summary, this study confirmed Annexin A2 and CDKN2B as new curcumin-binding proteins in oral squamous carcinoma cells.

Objective The aim of this study was to explore the effects of Tideglusib, a specific chemical inhibitor of GSK3β, on proliferation and functional gene expression of mouse osteoblast MC3T3-E1. Methods The proliferation of mouse osteoblast MC3T3-E1 was detected by CCK-8 assay after culturing in gradient concentrations ( 0, 10, 20, 40, 60, 80, 100, 1000, 2000 ng/L) of Tideglusib for 1, 2 and 3 days respectively to select the effective concentration. The adhesion and spreading morphology of MC3T3-E1 cells culturing in effective concentration (0, 10, 20 ng/L) of Tideglusib for 8 h was observed by fluorescence microscopy. The mRNA expression of Runx2, OSX, ALP, OPN and OCN of MC3T3-E1 cells was analyzed by real-time PCR and the protein expression of Runx2, OCN of MC3T3-E1 was detected by Western blot assay after culturing by 0, 10, 20 ng/L Tideglusib for 7 and 14 days． Results Compared with the control group, Tideglusib could promote cell proliferation and adhesion of mouse osteoblast MC3T3-E1 at the effective concentration of 10, 20 ng/L, and the effect was stronger at 20 ng/L. The mRNA level of Runx2, OSX, ALP, OPN and OCN was higher than that under 0 ng/L, and the protein level of Runx2, OCN also enhanced, which are more pronounced at 20 ng/L. Conclusions The specific chemical inhibitor of GSK3β, Tideglusib, can promote the proliferation and functional gene expression of osteoblasts.

[Abstract] Objective: To investigate the influence of miR-7704 expression on the biological behavior of head and neck squamous cell carcinoma (HNSCC) cells. Methods: The discrepant expression levels of miR-7704 were detected by real-time quantitative RT-PCR (qRT-PCR) among human oral epithelial cell line HOK and HNSCC cell lines, HN6, HN13, HN4 and CAL27. The expression level of miR-7704 was confirmed by qRT-PCR when HN6 and CAL27 was transfected with miR-7704 mimic and mimic NC. And, the effects of miR-7704 overexpression on the proliferation, migration and invasion of HNSCC cells were investigated by using CCK8, plate colony formation, wound healing experiment and Transwell invasion experiment. At the same time, total cell proteins were extracted, and the expression levels of EMT related indicators and threonine kinase (AKT) signaling pathway were detected by Western blot after the increase of miR-7704 expression. Results: Compared with HOK cell line, the expression levels of miR-7704 in all HNSCC cell lines were significantly up-regulated. After transfection of HNSCC cells with an overexpressed plasmid miR-7704 mimic, the expression level of miR-7704 was increased. Meanwhile, the cell proliferation, migration and invasion ability were enhanced. Western blot results showed that the expression levels of e-cadherin and beta-catenin in HNSCC cells were decreased after the overexpression of miR-7704, when the up-regulated expression of vimentin, Snail and Slug protein was observed. Levels of threonine kinase (AKT) and phosphorylated threonine kinase (p-akt) were significantly elevated. Conclusion: miR-7704 may act as a pro-oncogenic factor and induce EMT by regulating the AKT signaling pathway to promote the biological behavior of HNSCC cells.

Objective: To evaluate the bony and dental characteristics of mandibular second molar impaction and to provide reference for clinical treatment. Methods: Pretreatment CBCT records of 53 subjects of impacted mandibular second molar were selected as impaction group, and the same number of fully erupted mandibular second molar as eruption group. Mimics 19.0 software was used to observe and measure the indexes like SNA, SNB, ANB, Co-Gn, Co-Go, Go-Gn, W, Co-Go-Gn, retromolar space, tooth size and posterior tooth crowd. Independent t test and binary linear regression were analyzed and significant difference was identified with P＜0.05. Results: The impaction and eruption group didn’t differ significantly in respect to the bony measurements. Retromolar space was significantly shorter, and second molar was wider in the impaction group. Posterior tooth crowd of third molar in impaction group was statistically bigger than in the eruption group. Conclusions: Mandibular second molar impaction seemed to be relevant to retromolar space, mesiodistal width of the second molar, and the posterior tooth crowd.

[Abstract]Objective:In view of the Ann Ⅱ classes for children with mandibular retraction correct implementation of the muscle power training and analyzes its application effect. Methods: Selected from January 2018 to March 2019 in our hospital for treatment of 65 cases of Ann Ⅱ class children with mandibular retraction as the research object, all wearing muscle training, on the basis of the cooperate with muscle function training in orthodontic treatment. Clinical observation indexes such as SNA, SNB, ANB and anterior tooth coverage level before and after treatment were analyzed, as well as the compliance of children and the satisfaction of parents on correction.Results:Before correction, the coverage levels of SNA, SNB, ANB and anterior teeth were (85.12±6.39) °, (86.13±6.87) °, (5.37±1.24) ° and (6.38±1.25) mm, respectively. After correction, the levels of SNA, SNB, ANB and anterior tooth coverage were (81.31±4.20) °, (79.10±3.34) °, (3.45±0.71) ° and (3.27±0.64) mm, respectively, which were significantly improved after correction compared with those before correction, with no statistical significance (P > 0.05). The compliance at 1 week of correction was 87.68%(57/65), and 96.92%(63/65) at 12weeks of correction. The compliance at 12 weeks of correction was higher than that at 1 week, and the difference was statistically significant (P<0.05). At 1 week of correction, parents' total satisfaction on the correction effect was 84.62%(55/65), and at 12 weeks of correction, parents' total satisfaction on the correction effect was95.38%(62/65). The compliance at 12 weeks of correction was higher than that at 1 week, and the difference was statistically significant (P<0.05).Conclusion: Muscle function training application in Ann Ⅱ class mandibular retraction in orthodontic treatment, can effectively improve phase for teeth second deep deep coverage and mandibular retraction, adherence to rectify the ascending children, the parents of fully endorsed, has high clinical value, is worth popularizing widely. [Key words] Muscle trainer; Muscle work training; Ann Ⅱ classes; Ankylosis of the jaw; Clinical curative effect

Periodontitis is a chronic infectious disease with a high incidence rate. Scholars have paid great attention to the study of its immunological mechanism. Mitochondrial DNA (mtDNA) is a major member of damage-associated molecular patterns (DAMPs) and can be involved in activating the innate immune response. In recent years, the relationship between mtDNA and inflammation has become a research hotspot. There are evidences showing mtDNA is involved in the occurrence and development of periodontitis. Besides, many evidences of mtDNA and inflammatory signaling pathways related to periodontitis suggest that mtDNA may play an important role in the inflammation of periodontitis. This review aims to provide clues for the further exploration of the relationship between mtDNA and periodontitis.

There are many kinds of bacteria in the oral microenvironment, and the coexistence of bacteria is realized by the form of biofilm between bacteria and bacteria. The LuxS/AI-2 quorum sensing system is present in a variety of Gram-positive and negative bacteria. By integrating specific quorum sensing signals, bacteria can distinguish between autologous and non-autologous, thereby initiating the expression of bacterial-related genes, further precisely regulating biological behaviors to adapt environment, such as bioluminescence, release of virulence factors, and biofilm formation. In this review, we focus on the generation of AI-2, regulation of quorum sensing system and related research in oral pathogens.