p16 and p53 can Serve as Prognostic Markers for Head and Neck Squamous Cell Carcinoma.

OBJECTIVE: The present study aimed to explore the expression and clinical significance of human papilloma virus-related pathogenic factors (p16, cyclin D1, p53) in patients with head and neck squamous cell carcinoma (HNSCC) and construct a predictive model. METHODS: The Cancer Genome Atlas was used to obtain clinical data for 112 patients with HNSCC. Expression of p16, p53, and cyclin D1 was quantified. We used the survival package of the R program to set the cut-off value. Values above the cut-off were considered positive, while values below the cut-off were negative. Kaplan-Meier analysis and univariate and multivariate Cox regression analyses were performed to investigate prognostic clinicopathological indicators and the expression of p16, p53, and cyclin D1. A predictive model was constructed based on the results of multifactor Cox regression analysis, and the accuracy of the predictive model was verified through final calibration analysis. Follow-up of patients with HNSCC at the Affiliated Hospital of Binzhou Medical University was conducted from 2015 to 2017, and reliability of the predictive model was validated based on follow-up data and molecular expression levels. RESULTS: According to the results, expression of p16 and p53 was significantly associated with prognosis (P < .05). The predictive model constructed based on the expression levels of p16 and p53 was useful for evaluating the prognosis of patients with HNSCC. The predictive model was validated using follow-up data obtained from the hospital, and the trend of the follow-up results was consistent with the predictive model. CONCLUSION: p16 and p53 can be used as key indicators to predict the prognosis of HNSCC patients and as critical immunohistochemical indicators in clinical practice. The survival model constructed based on p16 and p53 expression levels reliably predicts patient prognosis.

A case of oral tuberculous ulcer and literature review.

Our purpose is to clearly diagnose the tongue and back tuberculosis ulcer through detailed medical history collection combined with examination, so as to provide certain experience for the diagnosis and treatment of oral tuberculosis.

[Effects of miR-31-5p on HIF-1α/BNIP3 signaling pathway and the expression of osteoblast-related factors of dental pulp stem cells].

PURPOSE: To investigate the effects of microRNA-31-5p (miR-31-5p) on the signal pathway of hypoxia inducible factor-1α (HIF-1α)/Bcl-2/adenovirus E1B 19-kDa-interacting protein 3(BNIP3) and the expression of osteoblast-related factors of dental pulp stem cells(DPSCs). METHODS: Human dental pulp stem cells (DPSCs) were cultured in vitro and divided into the control group (no transfection), mimic NC group (transfected with negative control-miR-31-5p), miR-31-5p mimic group (transfected with hsa-miR-31-5p mimic), siRNA NC group (transfected with nonsense siRNA) and miR-31-5p siRNA group (transfected with miR-31-5p siRNA).The expressions of miR-31-5p, HIF-1α, BNIP3, alkaline phosphatase(ALP) and Runt-related transcription factor-2(Runx2) mRNA in DPSCs were detected by real-time fluorescence quantitative PCR; the proliferation of DPSCs was detected by MTT; ALP activity of DPSCs was detected by ALP activity test kit; and the protein expressions of HIF-1α, BNIP3 and Runx2 in DPSCs were detected by Western blot. Statistical analysis was carried out with SPSS 24.0 software package. RESULTS: Compared with the control group and mimic NC group, the A value, ALP mRNA expression level and activity, Runx2 mRNA and protein expression levels of DPSCs in miR-31-5p mimic group were significantly lower (P＜0.05), ALP staining decreased significantly, and the expression levels of miR-31-5p mRNA, HIF-1α, BNIP3 mRNA and HIF-1α, BNIP3, Beclin1 protein were significantly higher (P＜0.05). Compared with the control group and siRNA NC group, the A value, ALP mRNA expression level and activity, Runx2 mRNA and protein expression levels of DPSCs in miR-31-5p siRNA group were significantly higher (P＜0.05), ALP staining enhanced significantly, and the expression levels of miR-31-5p mRNA, HIF-1α, BNIP3 mRNA and HIF-1α, BNIP3, Beclin1 protein were significantly lower(P＜0.05). CONCLUSIONS: MiR-31-5p may inhibit the expression of osteoblast-related factors of DPSCs, and activating HIF-1α/BNIP3 signaling pathway.

Identification of autophagy-related biomarker and analysis of immune infiltrates in oral carcinoma.

BACKGROUND: Autophagy plays a vital role in the progression of the tumor. We aimed to investigate the expression, prognostic value, and immune infiltration of autophagy-related genes in oral carcinoma via bioinformatics analysis. METHODS: The microarray datasets (GSE146483 and GSE23558) of oral carcinoma were downloaded from Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between normal and diseased groups were identified by the Limma package. The screened autophagy-related gene was further validated by the human protein atlas (HPA) database, TCGA database, and GSE78060 dataset. RESULTS: A total of 18 upregulated (top 10: EGFR, TNF, FADD, AURKA, E2F1, CHEK1, BRCA1, BIRC5, EIF2AK2, and CSF2) and 31 downregulated (top 10: MAP1LC3A, PARK2, AGT, IGF1, TP53INP1, CXCL12, IKBKB, SESN1, ULK2, and RRAGD) autophagy-related (DEGs) were identified, and FADD was found to be related to the prognosis of oral cancer patients. Gene set enrichment analysis indicated that FADD-associated genes were significantly enriched in immune-related pathways. Moreover, correlation analysis revealed that FADD expression was associated with immune infiltrates. Upregulation of FADD is associated with poor survival and immune infiltrates in oral cancer. CONCLUSION: We speculated that FADD is involved in the immune regulation of oral cancer, as well as autophagy.

BCCIPß facilitates p53 ubiquitination via binding with E6 protein in high-risk HPV positive head and neck squamous cell carcinoma.

BRCA2 And CDKN1A Interacting Protein (BCCIP) is initially identified as a tumor suppressor. Some recent studies confirmed its p53 binding capability. In this study, we explored the regulatory effect of BCCIPß on p53 stability in HPV-positive and HPV-negative HNSCC cells. RNA-seq data from TCGA-HNSC were extracted for transcript isoform analysis in HPV-positive and HPV-negative tumors. HPV16-positive UM-SCC-47 (SCC47) and UM-SCC-104 (SCC104) and HPV-negative SCC-9 (SCC9) and UM-SCC-1 (SCC1) cell lines were used as in vitro cell models. Results showed that BCCIPß was the dominant transcript in both HPV-positive and HPV-negative HNSCC cases. Knockdown of BCCIPß decreased p53 protein concentration in the two HPV-negative cell lines but increased p53 concentration in the two HPV-positive cell lines. BCCIPß inhibition increased proliferation and G1/S transition of SCC9 and SCC1 cells. In comparison, BCCIPß inhibition slowed proliferation and increased G1 arrest of SCC104 and SCC47 cells. BCCIPß inhibition prolonged the half-life of p53 protein and reduced p53 ubiquitination in the two HPV16-positive cell lines. Co-IP assay confirmed interactions among BCCIPß, HPV E6, and p53 in both SCC104 and SCC47 cells. In comparison, only the interaction between BCCIPα and p53 was confirmed in these two cell lines. Either E6 or BCCIPß inhibition reduced p53 ubiquitination and increased p53 concentration. However, inhibiting E6 and BCCIPß at the same did not generate synergistic effects. On the contrary, p53 ubiquitination level was even higher in the combination group, with lower p53 concentration compared to the shE6 group. BCCIPß overexpression in SCC47 cells with HPV E6 depletion significantly reduced p53 ubiquitination. In conclusion, this study found a novel interaction between HPV E6 and BCCIPß in HPV16-positive HNSCC cells. The presence of HPV E6 turned BCCIPß from a p53 stabilizer to a ubiquitination facilitator. This mechanism helps explain why BCCIPß acted as a tumor suppressor in HPV-negative HNSCC but exerted oncogenic function in HPV16-positive HNSCC.

VCAM1-targeted RNA interference inhibits the proliferation of human oral squamous carcinoma HN12 cells.

In the present study, RNA interference (RNAi) was used to investigate the effect of vascular cell adhesion molecule 1 (VCAM1) silencing on the proliferation of human oral squamous carcinoma HN12 cells. HN12 cells were divided into three groups: The untreated blank control cell group (CK), the negative control group transfected with non-homologous vector (NC) and the positive group transfected with the target sequence VCAM1 small hairpin RNA (KD). Reverse-transcription polymerase chain reaction and western blot analysis were used to examine the effects of VCAM1-knockdown on the mRNA expression of VCAM1 and subsequent protein expression. Furthermore, the HN12 cell growth inhibition rate was detected using the cell counting kit-8 method. The VCAM1-targeted lentiviral vector RNAi significantly inhibited VCAM1 mRNA, and subsequent protein, expression. Compared with the NC group, the VCAM1 gene knockdown efficiency was ~85%, while the expression level of VCAM1 protein was reduced by ~74% in KD group cells. In addition, cell growth was significantly inhibited in the KD group, with a growth inhibition rate of ~34%. Therefore, this targeted lentiviral vector RNAi effectively inhibited VCAM1 gene, and subsequent protein, expression, as well as the proliferation of oral squamous carcinoma cells. These results may provide an experimental reference for the diagnosis and treatment of oral squamous cell carcinoma.

Association of Decreased Expression of Serum miR-9 with Poor Prognosis of Oral Squamous Cell Carcinoma Patients.

BACKGROUND: Dysregulation of miR-9 is a common feature of many types of cancers, including oral squamous cell carcinoma (OSCC). However, whether the expression level of serum miR-9 is changed in patients with OSCC remains unknown. MATERIAL/METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression level of serum miR-9 in OSCC patients, oral leukoplakia (OLK) patients, and healthy volunteers, then we evaluated the association between serum miR-9 expression level and clinical outcome of OSCC patients. RESULTS: The expression level of serum miR-9 was significantly downregulated in patients with OSCC or OLK in comparison with healthy controls (P<0.01). Serum miR-9 expression level was associated with various clinicopathological parameters, including T stage (P=0.013), lymph node metastasis (P=0.002), and TNM stage (P=0.007). In addition, the OSCC patients in the low serum miR-9 expression group had poorer overall survival rate (P=0.022) and disease-free survival rate (P=0.004) compared with those in the high serum miR-9 expression group. Multivariate analysis showed that serum miR-9 was an independent prognostic factor for OSCC. CONCLUSIONS: Serum miR-9 was downregulated in patients with OSCC and patients with OLK. In addition, low serum miR-9 was correlated with poor prognosis of OSCC, indicating miR-9 might play a tumor suppressive role in OSCC and can serve as a promising biomarker for this deadly disease.

[Dental pulp stem cells modified by HIF-1α can differentiate into blood vessels].

PURPOSE: To investigate the method of differentiating dental pulp stem cells (DPSCs) modified by HIF-1α into blood vessels. METHODS: DPSCs were extracted from teeth samples from 20 patients and were identified by Strol-1 and CD146. DPSCs were divided into experimental group and control group according to DPSCs were modified by HIF-1α not or. HIF-1α-mRNA expression was detected by RT-PCR. HIF-1α, VEGF, SDF-1, Ang-2 and PDGF expression were detected using Western blot in different time after culture for 1 d, 4 d, 7 d and 14 d. Statistical analysis was carried out with SPSS 16.0 software package. RESULTS: Most DPSCs appeared round, oval under phase-contrast microscopy. CD146 and Strol-1 showed green fluorescence. HIF-1α and HIF-1α-mRNA expression became higher with time passing and the difference was statistically significant (P<0.05). Compared with the control group, HIF-1α protein and mRNA increased obviously in the experimental group 1d, 4d, 7d and 14d after transfection, and the difference was statistically significant (P<0.05). The level of VEGF, SDF-1, Ang-2 and PDGF in the control group was changed unconspicuously, and the expression was not different at different times (P>0.05). The level of VEGF, SDF-1, Ang-2 and PDGF in the exprimental group increased, and the difference was statistically significant between different time points(P<0.05). Compared with the control group, the level of VEGF, SDF-1, Ang-2 and PDGF in the experimental group was higher 1 d, 4 d, 7 d and 14 d after transfection, respectively, and the difference was statistically significant(P<0.05). CONCLUSIONS: DPSCs modified by HIF-1α gene can successfully induce vascular differentiation in vitro, which provides foundation for further angioplasty.

Infantile fibromatosis of the pterygomandibular space.

Infantile fibromatosis (IF) is a benign, nonmetastasizing but locally aggressive tumor. Pterygomandibular space is deep and obscure, and IF in this space is seldom reported. We describe a typical case of IF in the pterygomandibular space that occurred in a 3-year-old girl. Intraoral incisional biopsy revealed the diagnosis of IF, and surgical resection of the tumor was performed. We intend to heighten the awareness of this tumor and emphasize the pathologic features and identifications of IF and discuss the diagnosis and treatment of this disease. For the high recurrence risk, follow-up with clinical examination and magnetic resonance imaging in pediatric patients is recommended.

Intraosseous mucoepidermoid carcinoma of jaws: report of 24 cases.

OBJECTIVE: Intraosseous mucoepidermoid carcinoma of jaws is rare, and management of the disease remains poorly understood. The aim of this study was to assist the diagnosis and treatment of the tumor. STUDY DESIGN: The records of clinical data and follow-up information were collected from 1996 to 2010 and retrospectively analyzed for clinical features, surgical intervention, and prognosis. RESULTS: Of 24 cases, 15 were male, and the average age was 47.33 years. The clinical presentation of this tumor varied. For primary lesions and neck nodes, radical surgery was performed; radiotherapy or chemotherapy was administered after operation. The survival rate of all patients was 66.7%. The average survival period was 53.3 months after surgery. CONCLUSIONS: Diagnosis should be based on clinical and pathologic manifestations, surgery is the first choice for patient treatment, and radiotherapy may improve prognosis and therefore should be recommended in postoperative period.

Clinical management of pediatric aggressive fibromatosis involving the mandible.

BACKGROUND: Pediatric aggressive fibromatosis (AF) is a rare, benign tumor with locally infiltrative growth. Therefore, how to prevent reoccurrence while maintaining the mandible contour and continuity as much as possible is very important when the mandible is involved. PROCEDURE: We selected 10 pediatric patients with AF involving the mandible in our department between January 2001 and December 2011. Patient clinical data, including patient characteristics, symptoms at presentation, management, and treatment outcome, were reviewed. RESULTS: Patients' ages ranged from 3 to 16 years with six males and four females. According to imaging, there were three cases where the tumor adhered to the mandible; the periosteum was resected with the tumor, followed by cryotherapy or cauterization. Another five cases involved the destruction of the periosteum and the cortical plate. Resection of the involved mandible with the tumor was performed, and the margin was trimmed. In the last two cases, the tumor had invaded the whole ramus, and the immediate iliac graft was operated on after resecting the lesions. Neither radiotherapy (RT) nor chemotherapy was used. There was no recurrence or contour defect of the face. The function of the mandible was not affected, and only one case showed a slight limitation when opening the mouth. CONCLUSIONS: For the treatment of pediatric AF, we recommend complete tumor resection. As for the involved mandible, preserving the mandible contour and continuity as much as possible and providing adjunctive therapy, such as cryotherapy or cauterization, are vital. RT is not recommended.

Pleomorphic adenoma of the salivary glands in children and adolescents.

BACKGROUND: Pleomorphic adenomas (PAs) of the salivary glands are rare in children and adolescents. We reviewed the clinical manifestations, pathologic features, treatment, and prognosis of salivary gland PA in 90 children and adolescents. METHODS: Clinical data including age, sex, location, symptoms, the period of evolution of the symptoms, pathologic type, and surgical treatment were evaluated. RESULTS: The median age was 16.5 years old. There was a 1.43:1 female-to-male ratio. The parotid gland was the most commonly affected major gland (56.7%), and the palate, the most common site of minor salivary gland involvement. Ultrasonography and computed tomography were performed in most cases for diagnosis. Superficial parotidectomy or total parotidectomy with facial nerve dissection and preservation was the most common surgical procedure. Submandibular triangle dissection was applied to all submandibular PAs. Classic-type PA was the most common histologic subtype (43.3%). CONCLUSIONS: Salivary gland PAs in children and adolescents have different characteristics compared with their adult counterparts in regard to histologic subtype and location. Surgical removal is the best treatment option for PAs in children and adolescents.

Blood vessel formation in the tissue-engineered bone with the constitutively active form of HIF-1α mediated BMSCs.

The successful clinical outcome of the implanted tissue-engineered bone is dependent on the establishment of a functional vascular network. A gene-enhanced tissue engineering represents a promising approach for vascularization. Our previous study indicated that hypoxia-inducible factor-1α (HIF-1α) can up-regulate the expression of vascular endothelial growth factor (VEGF) and stromal-derived factor 1 (SDF-1) in bone mesenchymal stem cells (BMSCs). The angiogenesis is a co-ordinated process that requires the participation of multiple angiogenic factors. To further explore the angiogenic effect of HIF-1α mediated stem cells, in this study, we systematically evaluated the function of HIF-1α in enhancing BMSCs angiogenesis in vitro and in vivo. A constitutively active form of HIF-1α (CA5) was inserted into a lentivirus vector and transduced into BMSCs, and its effect on vascularization and vascular remodeling was further evaluated in a rat critical-sized calvarial defects model with a gelatin sponge (GS) scaffold. The expression of the key angiogenic factors including VEGF, SDF-1, basic fibroblast growth factor (bFGF), placental growth factor (PLGF), angiopoietin 1 (ANGPT1), and stem cell factor (SCF) at both mRNAs and proteins levels in BMSCs were significantly enhanced by HIF-1α overexpression compared to the in vitro control group. In addition, HIF-1α-over expressing BMSCs showed dramatically improved blood vessel formation in the tissue-engineered bone as analyzed by photography of specimen, micro-CT, and histology. These data confirm the important role of HIF-1α in angiogenesis in tissue-engineered bone. Improved understanding of the mechanisms of angiogenesis may offer exciting therapeutic opportunities for vascularization, vascular remodeling, and bone defect repair using tissue engineering strategies in the future.

Clinical management of peripheral ameloblastoma.

Peripheral ameloblastoma is a rare epithelial odontogenic tumor, limited to the soft tissues of the gingiva or oral mucosa. Peripheral ameloblastoma represents approximately 2% to 10% of all ameloblastomas. It is always considered to be benign, but occasionally it may be locally aggressive or with malignant potential. In this article, we report 3 new cases of benign peripheral ameloblastoma and further discuss the clinical management of this disease.

Clinical management of cervical ectopic thymus in children.

OBJECTIVE: Cervical ectopic thymus (CET) is an extremely uncommon etiology of a neck mass in an infant. The aim of this study was to study and analyze the clinical manifestations, management principles, and pathological diagnosis of CET. METHODS: From 1995 to 2010, a total of 25,237 cases of head and neck lesions were treated in the Department of Oral and Maxillofacial Surgery, Shanghai Ninth People's Hospital. Among these huge numbers of lesions, there were only 3 cases of pathological-diagnosed CET presenting as neck masses. These 3 rare cases were interesting, and here, we report their clinical management and pathological diagnosis. RESULT: Three patients had a pathological diagnosis of CET. Their ages ranged from 4 months to 4 years. Clinically, all 3 patients presented with a painless neck mass and received surgical resection. Pathological diagnoses are based on hematoxylin and eosin and immunohistochemical staining. CONCLUSION: Painless swelling or neck mass is the major complaint for CET. Radiologic imaging can help determine the extent of the mass and relationship with adjacent structures. Surgery with frozen section remains the main method for pathological diagnosis and management.

Solitary neurofibroma arising from the infratemporal fossa in a child.

Neurofibromas are derived from the nerve sheath and are commonly located in the head and neck region. They usually occur between the ages of 30 and 50 years. Neurofibromas arising from the infratemporal fossa are quite rare, especially in children. We describe a solitary neurofibroma arising from the infratemporal fossa in an 8-year-old boy who presented with a painless mass in his right cheek. Computed tomographic scan showed a soft-density, not well-circumscribed mass located in the right infratemporal fossa. The tumor was resected via the transmandibular approach with an excellent outcome. The histologic examination with immunohistochemical staining yielded the diagnosis of neurofibroma.

Does radiation-induced fibrosis have an important role in pathophysiology of the osteoradionecrosis of jaw?

Osteoradionecrosis of the mandible is a serious complication following radiation therapy with or without surgical intervention for malignancies of the head and neck. The acknowledged clinical presentation of osteoradionecrosis is pain, fistulae of mucosa or skin, complete devitalization of bone and pathological fractures. Radiation-induced fibrosis is an irreversible pathological process, which leads to damages in lung, skin, intestine, and pelvic cavity after radiotherapy. Studies have proved that radiation-induced fibrosis is involved in the pathological onset, development, maintenance of osteoradionecrosis and there is dose-effect relationship between them, so the authors hypothesize that radiation-induced fibrosis plays an important role in osteoradionecrosis. Studies need to perform to look for more efficient methods of managing and preventing the osteoradionecrosis.