Objective：The number and proliferation level of myoblasts derived from the unsegmented paraxial mesoderm are lower than those of the somite-derived myoblasts after muscle injury. This study aims to investigate the effect of Fgf8 activation on the tongue myoblasts (derived from the somite) and the masseter myoblasts (derived from the unsegmented paraxial mesoderm) in the craniofacial region. Methods：A mouse model (Myf5-cre; Rosa26R-Fgf8, RF8) that specifically activates Fgf8 in myogenic progenitor cells and a red-green dual-fluorescence mouse model called Myf5-cre;mT/mG were used to obtain mouse embryos of different days that were soon processed. Masson staining was performed to observe the morphology of tongue and masseter muscle. Immunohistochemistry was used to detect the expression of Myosin and MyoD in the muscle. BrdU labeling assay was used to assess the cell proliferation. Results：Masson staining showed that Myf5-cre; Rosa26R-Fgf8 mice had disordered tongue muscle fibers, but no significant changes in masseter muscle. Immunohistochemical results showed that the reduction of Myosin in tongue muscle was significantly greater than that of masseter muscle. The increase of MyoD in the genioglossus muscle was significantly greater than that of the masseter muscle, but the expression of MyoD in the intrinsic tongue muscle decreased dramaticly. BrdU labeling assay showed that the growth rate of the proliferation of genioglossus myoblasts was significantly greater than that of masseter muscle, but there was no significant change in the intrinsic tongue muscle. Conclusions：Fgf8 activation significantly promotes the proliferation of genioglossus myoblasts derived from the somite and masseter myoblasts derived from the unsegmented paraxial mesoderm and inhibits their differentiation. Its regulating effect on the genioglossus muscle is significantly greater than that of the masseter muscle. In addition, Fgf8 activation did not promote the proliferation of somite-derived intrinsic tongue myoblasts, but its inhibitory effect on the differentiation was also greater than that of masseter myoblasts

Objective:?To verify the distribution of Gli1+ cells in maxillary palatal suture, and to explore the role of Gli1+cells in osteogenesis induced by maxillary expansion, so as to provide experimental data to support the orthodontic treatment of skeletal malocclusion. Methods:?Gli1-LacZ transgenic mice were established to detect the distribution of Gli1+ cells in mid-palatal suture with or without maxillary expansion, HE and Masson staining were applied to evaluate the effect of maxillary expansion. In addition, immunofluorescence staining was used to observe the spatial relationship between Gli1+cells and the osteogenic marker Runx2 during maxillary expansion procedure. Results:?Gli1-Lacz transgenic mice were successfully established and heterozygotes and mutants were used in the following experiments. Compared to the non-expansion group, the palatal suture of the maxillary expansion group was significantly widened, with increase of bone formation and the expression of osteogenic marker Runx2. Moreover, Gli1+ cells increased in maxillary expansion group and colocalized with the Runx2 expression. Conclusions:?Gli1+ cells in maxillary palatal suture participate in osteogenesis induced by maxillary expansion.

Objective:To explore the effect of reactive oxygen species (ROS) on the differentiation of tumor infiltrating T lymphocytes in oral squamous cell carcinoma (OSCC). Methods: The proportion of T lymphocyte subsets in tumor and distal normal tissue were analyzed by flow cytometry. ROS expression was analyzed with ROS kit. Genes associated with T cell metabolism (HIF1A, GLUT1, HK2) in carcinoma and distal normal tissues were quantified by real time qPCR. GEPIA2 database was used to analyze the differential expression of T cell HIF1A in tumor and distal normal tissues and the prognosis value of mRNA level of T cell HIF1A in patients. Results: The proportion of effectors T cells (CD45RA+ CCR7?) in cancer tissues was significantly higher than that in distal normal tissues (P<0.05). The abundance of ROS in CD3+ T cells in cancer tissues was higher than that in distal normal tissues (P<0.05), and in cancer tissues, the ROS was significantly accumulated in effector T cells up-regulated compared with its counterpart in effector memory T cells (P<0.05). There were no statistically significant differences of ROS between the effector T cell and effector memory T cell in the distal normal tissue (P>0.05). Expressions of HIF1A, GLUT1, HK2 in tumor-infiltrating T cells were higher in those in distal normal tissues (P<0.05). T cell HIF1A level was higher in tumor tissues than that in distal normal tissues, but no significant statistical difference was found. The HIF1A level of effector T cells in tumor tissues was significantly higher than that in distal normal tissues (P<0.05). Patients with higher expression of HIF1A in T cell have a better prognosis (P<0.05). Conclusions:The upregulation of ROS/HIF-1 signaling pathway in the OSCC tumor microenvironment was significantly correlated with the high proportion of effector T lymphocytes in tumor tissues.

Objective:? To explore the role of autophagy related genes (ATGs) in the pathogenesis of tongue cancer based on TCGA and HADb database, and to establish a prognosis prediction model of tongue cancer patients based on ATGs expression profile. Methods:?The differential expression of ATGs in tongue cancer was determined by Wilcox test, and the correlation between ATGs and overall survival rate was analyzed by Cox regression. Go and KEGG analysis revealed the signal pathway and biological process related to autophagy gene in tongue cancer. Finally, the relationship between ATGs and clinical characteristics of tongue cancer patients was evaluated. Results:? ATGs in tongue cancer and adjacent tissues were differentially expressed. Five differentially expressed genes (VEGFA, ATG9B, WDR45, BAK1, VAMP3) were significantly related to the overall survival rate of patients with tongue cancer, which could predict the prognosis of survival. Go and KEGG showed significant enrichment of several key autophagy related pathways, such as apoptosis, P53 signaling pathway, autophagy function, EGFR tyrosine kinase inhibitor resistance,etc. Three ATGs (ATG9B, VEGFA, WDR45) were significantly correlated with different clinicopathological features. Conclusions:?We identified five differential autophagy genes, VEGFA, ATG9B, WDR45, BAK1 and VAMP3, which can predict the prognosis of tongue cancer, and have reference value for autophagy research in the field of tongue cancer.

Objective:?To study the clinicopathological features, immunophenotype，differential diagnosis and molecular features of mammary analogue secretory carcinoma (MASC). Methods:?Microscope observation, immunophenotyping analysis and EVT6-NTRK3 genetic testing were performed on 4 cases of MASC which were diagnosed by the Department of Pathology, the First Affiliated Hospital of Nanjing Medical University and the Department of Pathology, Huai'an First People's Hospital from July to December 2018, and related literature was also reviewed. Results:?The patients were aged from 33 to 66 years old. The ratio of male to female was 1:1. Grossly, all cases revealed a solitary mass(1.3-2.2 cm in diameter). Microscopically, all cases demonstrated a highly infiltrative growth pattern. All tumors showed mixed growth patterns including microcystic, cystic-papillary, tubular, and solid, and eosinophilic secretions could be seen in the cavities. The mitoses were not easily found, and no coagulative necrosis, nerve or vascular invasion were found in these 4 cases. Immunohistochemically, all cases were positive for CK7, S-100, 乳腺球蛋白, SOX-10、GATA3. But DOG-1 and the other myoepithelial markers P63、α-SMA、Calponin were negative in all 4 cases. The Ki-67 proliferation index of 3 cases was less than 20%, and the other one case was about 35%. ETV6-NTRK3 gene rearrangement was detected in 3 cases by fluorescence in situ hybridization. Conclusions:?MASC is a rare low-grade salivary gland carcinoma exhibiting moderate morphology and diverse arrangement patterns. Its distinct histochemical feature - strong S-100, 乳腺球蛋白 positivity and DOG-1 negativity, can help to differentiate it from other low-grade saliva gland tumors. ETV6-NTRK3 gene rearrangement tests can be used as an auxiliary diagnostic tool if necessary.

Objective: To study the composition and function of the oral microbial flora of patients with periodontitis by metagenomic method combined with high-throughput sequencing technology, and explore the correlation between the composition and function of microorganisms and the occurrence and development of periodontitis. Methods: Saliva and subgingival plaque samples of 10 patients with periodontitis were collected, and total microbial DNA was extracted from each sample to build the macro-genome library, and MGISeq-2000 sequencing was conducted, while bioinformatics analysis was conducted using Metaphlan2 software and Humann2 software respectively. Results: Among the 20 samples, the microbial composition analysis results showed that at the genus level, the average relative abundance of Porphyromonas in the saliva and subgingival sample groups was not statistically different (P>0.05); the relative abundance of the Prevotella and Capnophagocytes were statistically different between the two sample groups (P<0.05). At the species level, the relative abundance difference of Haemophilus parainfluenzae and Neisseria flavus between the two sample groups was statistically significant (P<0.05). The result of functional analysis showed that the relative abundance of the two groups of contributing bacteria and sequences was statistically different (P<0.05). Conclusions: The results suggested that the subgingival microbe samples may be more representative for the study of periodontitis pathogenic bacteria and that the use of metagenomics to study metabolic pathways related to periodontitis disease may deserve further study.

Objective:?To study the effects of chronic sleep deprivation on the histology of temporomandibular joint (TMJ) in rats. Methods:?60 rats were randomly divided into control group and experimental group. The rats were then chronicly deprived of sleep for 4, 6 and 8 weeks by the modified multiple platform method (MMPM). Finally, the pathological changes, angiogenesis, and the expression of vascular endothelial growth factor (VEGF) of TMJ condylar cartilage were evaluated by hematoxylin and eosin (HE) staining, CD105 fluorescent staining, and immunohistochemistry, respectively. Results:?MMPM can induce the rats into chronic sleep deprivation state successfully. HE staining showed osteoarthritic (OA)-like pathological changes, such as fiber fracture, reduced chondrocytes, disordered arrangement, and increased capillaries in the experimental group, while no changes were observed in the control group. The osteochondral junction MVDs of the experimental group were significantly higher than that of the control group (P <0.05)，and showed increasing trend with the prolonged sleep deprivation. Immunohistochemisty showed that VEGF was highly expressed in the middle and even superficial layers of the condyle cartilage in the experimental group, while the expression was relatively lower in each layer of the condyle cartilage in the control group. Conclusions:?Sleep deprivation of rats through MMPM could induce angiogenesis of TMJ condylar cartilage and TMJ-OA like histological changes in rats, which showed progressive enhancement in a time-dependent manner.

Objective:?The three-dimensional finite element models with different shapes of the split post were established, and the von Mises stress distributions were analyzed. Methods:?According to the slope angle (90°, 120°, 135° and 160°) formed between the root post (the split post in root canal) and the root canal orifice and the diameter difference between the crown post (the part above the root canal) and the root post (0.5 mm, 1 mm, 1.5 mm and 2 mm), the three-dimensional finite element models of 16 different forms of split posts were established by SolidWorks 2016 and ANSYS software. The Von Mises stress distributions of different forms of split posts were analyzed under the conditions of 600 N vertical loading and 225 N horizontal loading, respectively. Results:?The stress distribution of 16 groups of split post models was basically similar under different directions of loading, and the maximum stress was mainly distributed at the junction of crown post and root post; when the root canal orifice was not prepared for slope, the maximum von Mises stress at any diameter difference was significantly higher than that of obtuse angle group with slope (120°, 135° and 160°) under horizontal and vertical loading; When the slope was 120°, the von Mises stress under any diameter difference was lower than that at 135°and 160°angles; When the angle of slope was greater than 90°, the stress at the joint of crown post and root post decreased with the increase of diameter difference, and when the diameter difference between crown post and root post was 2 mm, the von Mises stress value under horizontal and vertical loads was the minimum, regardless of the angle at the joint of crown post and root post. Conclusions:?The slope angle of the split post is recommended to be 90° to 120° in the split zirconia post-crown, and the larger the diameter difference between the crown post and the root post, the better the stress distribution at the joint between the crown post and the root post.

SNRP200 is the splicing gene of nuclear precursor mRNA, and the encoded protein belongs to DEXH-box protein family of RNA helicase, which is widely expressed in the whole body tissues. In recent years, studies have shown that SNRNP200 is closely related to the splicing of precursor mRNA, and it can regulate the expression of related genes by influencing splicing, thus affecting cell proliferation. In addition, SNRNP200 also plays an important role in the pathogenesis of hereditary retinitis pigmentosa and acute myeloid leukemia (AML). In this paper, the structural characteristics, functional regulation and effects on diseases of SNRNP200 were reviewed.