VIRMA promotes the progression of head and neck squamous cell carcinoma by regulating UBR5 mRNA and m6A levels.

Head and neck squamous cell carcinoma (HNSCC) is a globally prevalent and lethal cancer form, whose precise mechanisms remain incompletely understood. Increasing evidence suggests that N6-methyladenosine (m6A) plays a crucial role in cancer progression. This study aimed to explore the biological function of m6A modification and vir-like m6A methyltransferase associated (VIRMA) in HNSCC. We conducted an analysis of VIRMA expression in HNSCC cells using The Cancer Genome Atlas (TCGA) database and employed reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting to assess its expression levels in HNSCC cell lines. Additionally, m6A levels in HNSCC cells were quantified, and the correlation between VIRMA expression levels and the clinical and pathological features of other genes was analyzed. Upon knocking down VIRMA levels, we assessed HNSCC cell proliferation, migration, and invasion and validated downstream genes using RT-qPCR and western blot. Our findings suggested that VIRMA, as an m6A-related regulator, may significantly influence HNSCC progression by regulating ubiquitin protein ligase E3 component N-recognin 5 (UBR5) through m6A modification. Therefore, VIRMA may serve as a prognostic biomarker.

Imaging analysis of 1 138 supernumerary teeth by using cone-beam computed tomography.

OBJECTIVES: This study aims to analyze and summarize the characteristics of supernumerary teeth by using cone-beam computed tomography (CBCT). METHODS: A total of 718 patients with 1 138 supernumerary teeth were retrospectively collected. Age, gender, number, location, morphology, eruption status, and accompanying symptoms of the supernumerary teeth were statistically analyzed. The relationship relative to jaws, gender, and eruption status were analyzed and discussed. RESULTS: The average age of the patients was 9.54±5.32 years, and the male to female ratio was 2.88∶1. About 77.02% of the patients sought medical advice during the mixed dentition period, and 50.70% had one supernumerary tooth. These supernumeraries were most commonly conical in shape, and 85.76% of them were in the incisor region, 92.09% in the upper jaw, 46.75% in inverted position, and 86.20% unerupted. Overall, 65.29% of them had fully developed roots, and 60.63% had an impact on adjacent structures. Significant differences were found in eruption status, morphology, zoning, direction, root development, and impact on adjacent structures between the supernumerary teeth located in the upper and lower jaws (P<0.05). Significant differences were also detected in gender, morphology, zoning, orientation, root development, and impact on adjacent structures between erupted and unerupted teeth (P<0.05). The incidence of supernumerary teeth in the incisor region was higher in males than that in females. Moreover, the root of supernumeraries was more completely developed in males than in females (P<0.05). CONCLUSIONS: For supernumerary teeth, CBCT images can provide accurate three-dimensional radiographic data and are valuable for clinical diagnosis and treatment planning.

Inducing differentiation of human dental pulp cells toward acinar-lineage.

OBJECTIVE: To induce acinar-differentiation from human dental pulp cells for potential application in aiding treatment of dry-eye syndromes. METHOD: Human dental pulp cells were co-cultured with human submandibular gland acinar cells using a transwell construction for 2 weeks. The two populations of cells were physically separated while chemical and biochemical components can be exchanged. Fibroblasts were included as a negative control. Expression of amylase, cytokeratin 8 and vimentin were examined by immune-staining. Amylase activity was measured using an AMS Assay Kit. RESULT: Cobblestone-like islands, a feature of acinar cells, appeared in the dental pulp cells which were co-cultured with salivary gland cells for one week and increased in number and size after two weeks. Antibody detected amylase in 30 and 50% of the pulp cells 1 and 2 weeks in the co-culture, respectively. Cytokeratin 8 increased while vimentin decreased. All these changes indicate an acinar-like differentiation of the dental pulp cells. None of these changes were observed in fibroblasts which were also co-cultured with salivary gland cells, indicating that the acinar-like differentiation is specific for the dental pulp cells. Neither of the changes were observed in dental pulp cells when not co-cultured with the salivary gland cells, indicating that induction is specific and essential. CONCLUSIONS: Human dental pulp cells have the potential to differentiate into acinar-like cells which may provide an autologous source for cellular therapy for dry-eye syndromes.

Concentrated growth factor inhibits UVA-induced photoaging in human dermal fibroblasts via the MAPK/AP-1 pathway.

Ultraviolet (UV) radiation-induced photoaging is one of the contributors to skin aging. UV light triggers oxidative stress, producing a large number of matrix metalloproteinases (MMPs) and degrading the extracellular matrix in skin cells, thereby causing a series of photoaging symptoms. Concentrated growth factor (CGF) is a leukocyte- and platelet-rich fibrin biomaterial that plays a protective role in the occurrence and development of skin photoaging. In the present study, we investigated the underlying mechanism of CGF in the UVA-induced photoaging of human dermal fibroblasts (HDFs). A primary culture of HDFs was isolated from normal human facial skin. The cells were treated with CGF following UVA radiation. Proliferation of cells was detected using MTT assay, followed by measurement of reactive oxygen species (ROS) using immunofluorescence assay and flow cytometry. The mRNA and protein expression levels of P38, c-Jun, and MMP-1 were detected using real-time polymerase chain reaction and Western blot, respectively. CGF was found to improve cell viability by inhibiting the production of ROS and reducing oxidative damage. In addition, there was lower expression of p38 and c-Jun at the mRNA and protein levels following CGF treatment, thus resulting in the inhibition of MMP-1 expression. Our results suggest that CGF could protect HDFs against UVA-induced photoaging by blocking the P38 mitogen-activated protein kinase/activated protein-1 (P38MAPK/AP-1) signaling pathway. These findings provide a new clinical strategy for the prevention of skin photoaging.

Concentrated Growth Factors Can Inhibit Photoaging Damage Induced by Ultraviolet A (UVA) on the Human Dermal Fibroblasts In Vitro.

BACKGROUND Photoaging is the main cause of extrinsic skin aging. Daily exposure to ultraviolet A (UVA) accelerates the process of photoaging. The present study aimed to understand the role of concentrated growth factors (CGF) on UVA irradiated human skin cells. MATERIAL AND METHODS We isolated and subcultured normal human dermal fibroblasts (NHDFs) from 6 different human dorsal skins and established photoaging models of NHDFs irradiated by UVA to detect the influence of CGF on fibroblasts in vitro. Three groups were examined: normal, cellular photoaging model (total dosages of 18J·cm--⁻²-), and cellular photoaging model plus CGF. In our study, we used the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay method to measure the cell viability. We also used reactive oxygen species (ROS) assay and superoxide dismutase (SOD) assay to measure respectively the amount of oxygen free radicals and antioxidative enzymes. We compared the migration rates among the photoaging model groups, the control groups, and the CGF-treated culture medium groups that were irradiated. RESULTS Our study results indicated that 5% CGF can reduce UVA-induced human skin fibroblasts damage significantly, improve the viability of NHDFs significantly, and largely decrease the UVA irradiation effect (P<0.05). The migration rates of the normal group and the UVA-irradiated NHDFs in the 5% CGF group had significantly increased migration rates (P<0.05), compared to the control medium group. The migration rates of the UVA-irradiated NHDFs in 5% CGF exceed those of the normal group. These results showed that 5% CGF could greatly promote cellular proliferation, migration, and SOD at the same time that the amounts of ROS were markedly decreased. CONCLUSIONS These experimental findings offer some important insights into CGF's capacity for scavenging ROS, improving SOD, and increasing migration rates in NHDFs irradiated by UVA.

Effect of Xylosyltransferase-I Silencing on Implanting Growth of Salivary Pleomorphic Adenoma.

BACKGROUND Salivary pleomorphic adenoma is one of the most common salivary gland tumors. It has a relatively high tendency to recur and a high risk of malignant transformation. The present study aimed to study the effect of XT-I gene silencing on the implanting growth of salivary pleomorphic adenoma. MATERIAL AND METHODS Primary cultures of SPA cells and fibroblasts from the same patient were assessed. The adenovirus vector Ad-shRNA-XT-I was constructed and transfected into SPA cells. The expression of XT-I gene and XT-I protein was detected by real-time PCR and Western blot. The contents of proteoglycans were detected. The SPA cells transfected with Ad-shRNA-XT-I (group SPA-XT-I) and Ad-shRNA-HK (group SPA-HK), as well as without transfection (group SPA), were implanted into ADM scaffold with fibroblasts and then transferred into 18 BALB/C-nu nude mice for 3 months. RESULTS Primary cultures showed SPA cells were positive for human CK and S-100 protein and the fibroblasts were positive for human vimentin. The expressions of XT-I gene and protein were decreased by 51% and 51.31%, respectively. The content of proteoglycans was reduced by 48.45%. The results of the implanting growth in vitro and in vivo of nude mice indicated that no tumors grew in the SPA-XT-I group, whereas SPA grew in groups SPA-HK and SPA positive for human a-SMA, S-100 protein, and calponin. CONCLUSIONS XT-I gene silencing effectively inhibited the implanting growth of SPA.

Morphological Observation on Critical-Sized Cranial Defect Repaired by Icariin and Autologous Concentrate Growth Factors in Rabbits.

BACKGROUND Morphological changes repaired by icariin and autologous concentrate growth factors (ACGF) in critical-sized cranial defect were observed and their promoting effects were investigated. MATERIAL AND METHODS Seventy-two New Zealand white rabbits weighing 1.8~2.0kg were used to build a critical-sized cranial defect model and were randomly divided into 3 groups. X-ray, HE staining, general and histological observation, and immunohistochemistry were used to describe the changes caused by normal saline, icariin, and ACGF. RESULTS Cranial defects were covered with newly formed bone tissue at the 12th week in icariin and ACGF groups, with red color, hard surface, and no obvious boundary. Densities were the same in 2 groups at 4 timepoints. HE staining showed defects filled with a large amount of fibrous connective tissue, thick collagen fibers, and abundant osteoclasts. No new bone matrix appeared in any of the 3 groups. Trabecular area, trabeculae width, and osteoblast number in 2 groups were more than that of the control group, and osteoclast number was lower. However, osteoclast number among the 3 groups at the 12th week had no significant difference, which was the same with 4 indicators between the icariin and ACGF groups. From the 4th to 12th week, regenerated cartilage was formed and showed positive reaction with BMP-2 and TGFß1 from primary bone, which also was demonstrated by granulation tissue and uniform dyeing. CONCLUSIONS ACGF and icariin both can increase new bone quantity and improve bone quality, which can also promote healing. The effects and mechanisms of icariin and ACGF on the expression of gene are not exactly the same.

The relationship between proteoglycan inhibition via xylosyltransferase II silencing and the implantation of salivary pleomorphic adenoma.

OBJECTIVE: To study the relationship between proteoglycan inhibition and the implantation of salivary pleomorphic adenoma (SPA). METHODS: SPA fresh tissue was primitively cultured and identified. The Ad-shRNA-XT-II adenovirus vector was constructed and transfected into SPA cells to silence the XT-II gene. The expression of the XT-II gene and protein was detected using real-time PCR and Western blotting, respectively. The proteoglycan content of the cells was determined 48 h after transfection. The invasion and migration of SPA cells were observed using Matrigel invasion and wound-healing assays. Fibroblasts from the tumour capsule of the same patient were obtained, cultured and seeded onto an acellular dermal matrix (ADM) scaffold. Tumour cells were seeded onto the scaffold with the fibroblasts and then transferred to BALB/C-nu nude mice and allowed to grow in vivo for 3 months. RESULTS: The SPA cells cultures were positive for human calponin, S-100 protein, α-SMA and CK. XT-II gene and protein expression was decreased by 42.72% and 34%, respectively. The proteoglycan content was downregulated by 41.15%. XT-II gene silencing decreased the invasion and migration abilities of SPA cells. The assessment of SPA growth in nude mice indicated an absence of tumour growth in the SPA-XT-II group (in which the XT-II gene was silenced), whereas SPA growth was observed in the other two groups (in which the XT-II gene was not silenced), and the tumour tissue was positive for the human S-100 protein, α-SMA and CK8&18. CONCLUSION: Proteoglycan inhibition induced via XT-II gene silencing inhibited the implantation of SPA.

[Rapid canine distalization through distraction of the periodontal ligament after reducing interseptal bone resistance].

OBJECTIVE: To investigate the effect of rapid canine distalization through distraction of the periodontal ligament after reducing interseptal bone resistance. METHODS: Twenty canines in 11 patients who needed first premolar extractions were involved. A tooth-borne, custom-made distractor was bonded right after the first premolar extraction and the interseptal bone resistance reduction. Three days post-operatively, the distractor was activated 0.1 mm three times a day. Orthodontic models, panoramic radiographs, periapical radiographs, electrical vitality test were assessed pre- and post distraction procedure and 3 months after the completion of the procedure. RESULTS: The distraction procedure was completed in 18 to 35 days [mean (25.6 +/- 4.7) days], with the distal displacement of the canines ranging from 3.53 to 8.29 mm [mean (5.56 +/- 1.32) mm]. The canines showed a mean of 12.20 degrees distal tipping and 18.53 degrees rotation. The anchorage teeth showed an average of (0.76 +/- 0.75) mm mesial movement. The mesial contact point of incisors showed a mean of (0.67 +/- 0.55) mm lingual movement. There was no significant root resorption or long-time change on pulp vitality after distraction. CONCLUSIONS: The canine distalization through distraction of the periodontal ligament after reducing interseptal bone resistance was an effective approach to move canines rapidly.

[Study on effect of snore guard to upper airway structure of normal occlusion people by magnetic resonance imaging].

OBJECTIVE: To investigate the imagery changes of the upper airway and the surrounding soft tissues of local adults with non-apnea who used snore guard and to provide experimental data for the clinical diagnosis and treatment of obstructive sleep apnea syndrome (OSAS). METHODS: Thirty students with non-apnea from Hebei medical university were chosen, and magnetic resonance imaging (MRI) was used to measure the changes of the upper airway and the surrounding soft tissues after snore guards were used. SPSS 105 software was used to analyze statistically. RESULTS: After the snore guard was put into oral cavity, the change of the average section and volume of the nasopharynx, the palatopharynx, the hypopharynx and the glossopharynx were statistically significant. The average sagittal size, the average horizontal size of the nasopharynx, the palatopharynx, the hypopharynx and the glossopharynx were increased statistically. The ratio of sagittal size, the horizontal sizand the in the hypopharynx and the glossopharynx changed statistically important. There was a decrease of the soft palate, the shape, the height, and the length of the tongue, the difference was statistically significant. The results demonstrated that snore guard affected the upper airway mainly by changing the volume and the shape of the upper airway, there was an obvious increase of the pharynx. The results also showed that snore guard could increase the width (both sagittal and horizontal) of the upper airway and could change the shape of the surrounding soft tissues, which caused air way more smooth. Snore guard could make the indexes of soft palate and tongue change decreasingly, resulted in the straight stand up of the tongue and the forwardness of the soft palate. CONCLUSION: Snore guard is an effective and convenient instrument for treating the patients with OSAS.

[The effect of hyaluronic acid external film on rats wound healing].

OBJECTIVE: To observe the effect of an external film of hyaluronic acid (HA) on the rats wound healing. METHODS: Forty-eight SD rats were randomly separated into eight groups of 6 rats each. Bilateral dorsal cuts were performed on each rat, left wound was used as the experiment with HA external film and right wound was used as the control only with normal saline. The process of healing was observed histologically following 1st, 3rd, 5th, 7th, 9th, 14th, 21st, and 28th days postoperatively. RESULTS: Inflammation was lighter and epidermal healing was faster in the experimental group than those in the control. The fibroblasts degenerated and the collagen fiber changed to slim and loose bunches in the experimental group. CONCLUSION: The results indicated that HA external film could have powerful infiltrating activity at the early stage of wound healing, it could accelerate the healing of epidermis and delay the formation of keratinization layer.