A splicing mutation in VPS4B causes dentin dysplasia I.

BACKGROUND: Dentin dysplasia I (DDI) is a genetically heterogeneous autosomal-dominant disorder characterised by rootless teeth with abnormal pulpal morphology, the aetiology of which presents as genetically heterogeneous. METHODS AND RESULTS: Using a cohort of a large Chinese family with 10 patients with DDI, we mapped to a 9.63 Mb candidate region for DDI on chromosome 18q21.2-q21.33. We then identified a mutation IVS7+46C>G which resulted in a novel donor splice site in intron 7 of the VPS4B gene with co-segregation of all 10 affected individuals in this family. The aberrant transcripts encompassing a new insert of 45 bp in size were detected in gingival cells from affected individuals. Protein structure prediction showed that a 15-amino acid insertion altered the ATP-binding cassette of VPS4B. The mutation resulted in significantly reduced expression of mRNA and protein and altered subcellular localisation of VPS4B, indicating a loss of function of VPS4B. Using human gingival fibroblasts, the VPS4B gene was found to act as an upstream transducer linked to Wnt/ß-catenin signalling and regulating odontogenesis. Furthermore, knockdown of vps4b in zebrafish recapitulated the reduction of tooth size and absence of teeth similar to the tooth phenotype exhibited in DDI index cases, and the zebrafish mutant phenotype could be partially rescued by wild-type human VPS4B mRNA. We also observed that vps4b depletion in the zebrafish negatively regulates the expression of some major genes involved in odontogenesis. CONCLUSIONS: This study identifies VPS4B as a disease-causing gene for DDI, which is one of the important contributors to tooth formation, through the Wnt/ß-catenin signalling pathway.

Reconstruction of Anterior Mandibular Defect Using Submental Island Flap Pedicled With Mental Artery.

Gingival carcinoma is a common malignant tumor occurring in the anterior area of the mandible, which can be derived from the epithelium of gingival mucosa. Surgical extended resection is the main treatment of gingival cancer, which can lead to anterior mandibular defect including mouth floor and mandible and mucosa of lower lip. According to the size of the defect, the common repair method is free musculocutaneous flap with vascular pedicle or pedicle flap. We present a method of repairing mandibular anterior tooth defect with an island flap pedicled with the mental artery.

The occurrence, severity degree, and associated risk factors of dental fluorosis among the 12-year-old schoolchildren in Jilin, China.

ABSTRACT: This study aims to describe the occurrence, severity degree, and correlated risk factors of dental fluorosis among the 12-year-old schoolchildren of Jilin, China.We conducted a cross-sectional, observational, and descriptive study among 960 12-year-old schoolchildren in Jilin. The Dean index was utilized to evaluate the severity degree of dental fluorosis. A questionnaire was sent to the guardians of children. Community fluorosis index was measured to estimate the importance of enamel fluorosis for the whole population's public health. The logistic regression analysis was also utilized to identify the correlation between fluorotic teeth and the independent variables.Nine hundred sixty children were assessed. Among them, 480 (50%) were female. 30.5% of subjects had dental fluorosis, 7.19% had very mild dental fluorosis, 10.73% experienced mild dental fluorosis, 9.58% suffered moderate dental fluorosis, and 3.02% encountered severe dental fluorosis. The overall community fluorosis index was 0.73. The results of logistic regression showed that schoolchildren who brushed teeth more frequently (OR: 2.012, 95% CI 1.767-2.342), deficiency of parental supervision (OR: 4.219, 95% CI 3.887-4.573), and lived in rural areas (OR: 2.776, 95% CI 2.163-3.489) were more correlated with enamel fluorosis. Moreover, schoolchildren whose mothers or fathers were of high education level (OR: 0.336, 95% CI 0.217-0.413 and 0.346, 95% CI 0.113-0.512) and only child (OR: 0.378, 95% CI 0.213-0.415) were protective factors for dental fluorosis.In the Jilin province of China, the risk indicators for dental fluorosis include rural areas, more frequency of brushing, low educational background of parents, and deficiency of parental supervision.

Multidecadal records of microplastic accumulation in the coastal sediments of the East China Sea.

Microplastics are an emerging hazard in the marine environment, and considered to eventually sink into sediments. An investigation into the long-term variation of microplastic accumulation in sediment cores is essential for understanding the historical trend of this contamination and its response to human activities. In this study, the multidecadal changes of microplastic abundances in two sediment cores from the inner shelf of the East China Sea (ECS) were revealed by two methods, i.e., a visual enumeration method based on scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS) and a quantitative method based on microplastic-derived carbon (MPC) abundances. The features of microplastics were determined via SEM-EDS and micro-Fourier transform infrared spectroscopy (µ-FTIR). The results reveal a multidecadal increasing trend of microplastic accumulation in the coastal sediments of the ECS since the 1960s, which may be jointly governed by the release of plastic wastes and oceanographic dynamics. Meanwhile, the breakpoint of the exponential growth of microplastics in the ECS occurs in 2000 AD, which well matches the rapid increasing of plastic production and consumption in China. Further, based on the MPC contents in sediments, the influence of microplastics on the quantitative evaluation of carbon storage in the ECS has been examined for the first time, revealing an insignificant (<2% before 2014 AD) but potentially-increasing (6.8% by 2025 AD) contribution of microplastics to carbon burial. Our results may provide the important data for evaluating and mitigating the impact of microplastics on the marine environment.

Antitumor effect of TRAIL on oral squamous cell carcinoma using magnetic nanoparticle-mediated gene expression.

We developed a new magnetic nanovector to improve the efficiency and targeting of transgene therapy for oral squamous cell carcinoma (OSCC). Positively charged polymer PEI-modified Fe(3)O(4) magnetic nanoparticles were tested as gene transfer vectors in the presence of a magnetic field. The Fe(3)O(4) nanoparticles were prepared by a co-precipitation method and had good dispersibility in water. These nanoparticles modified by PEI were combined with negatively charged pACTERT-EGFP via electrostatic interaction. The transfection efficiency of the magnetic nano-gene vector with the magnetic field was determined by a fluorescence-inverted microscope and flow cytometry. The results showed significant improvement compared with the control group (p < 0.05). The magnetic complexes also exhibited up to 6-times higher transfection efficiency compared with commonly used PEI or lipofectin. On the basis of these results, the antitumor effect with suicide gene therapy using pACTERT-TRAIL in vitro and vivo was evaluated. In vitro apoptosis was determined with the Annexin V-FITC Apoptosis Detection Kit. The results suggested that PEI-modified Fe(3)O(4) nanoparticles could mediate the killing of Tca83 cells. Furthermore, treatment with pACTERT-TRAIL delivered by magnetic nanoparticles showed a significant cytostatic effect through the induction of apoptosis in a xenograft model. This indicates that magnetic nano-gene vectors could improve the transgene efficiency for Tca83 cells and could exhibit antitumor functions with the plasmid pACTERT-TRAIL. This may be a new way to treat OSCC.

Differentiation of rabbit bone mesenchymal stem cells into endothelial cells in vitro and promotion of defective bone regeneration in vivo.

Tissue engineering strategies often fail to regenerate bones because of inadequate vascularization, especially in the reconstruction of large segmental bone defects. Large volumes of vascular endothelial cells (ECs) that functionally interact with osteoblasts during osteogenesis are difficult to obtain. In this study, we simulated bone healing by co-culturing differentiated ECs and mesenchymal stem cells (MSCs) either on a culture plate or on a polylactide glycolic acid (PLGA) scaffold in vitro. We also evaluated the effect of osteogenesis in repairing rabbit mandible defects in vivo. In this study, MSCs were separated from rabbit as the seed cells. After passage, the MSCs were cultured in an EC-conditioned medium to differentiate into ECs. Immunohistochemical staining analysis with CD34 showed that the induced cells had the characteristics of ECs and MSC. The induced ECs were co-cultured in vitro, and the induction of MSCs to osteoblast served as the control. Alkaline phosphatase (ALP) and alizarin red (AZR) staining experiments were performed, and the Coomassie brilliant blue total protein and ALP activity were measured. The MSCs proliferated and differentiated into osteoblast-like cells through direct contact between the derived ECs and MSCs. The co-cultured cells were seeded on PLGA scaffold to repair 1 cm mandible defects in the rabbit. The effectiveness of the repairs was assessed through soft X-ray and histological analyses. The main findings indicated that MSCs survived well on the scaffold and that the scaffold is biocompatible and noncytotoxic. The results demonstrated that the co-cultured MSC-derived ECs improved MSC osteogenesis and promoted new bone formation. This study may serve as a basis for the use of in vitro co-culturing techniques as an improvisation to bone tissue engineering for the repair of large bone defects.

Adenoid cystic carcinoma in the maxillary gingiva: a case report and immunohistochemical study.

Gingival adenoid cystic carcinoma (ACC) is a rare malignancy. We describe the diagnosis and treatment of a 43 year-old woman who presented with a persistent oral ulcer for approximately 1 year, and subsequent pain in the left posterior maxillary region. Clinical examination revealed an ulcer in the left upper molar gingiva, with swelling in the region from the second premolar to the third molar. X-ray images demonstrated the involvement of the maxillary alveolar bone. The histopathological and immunohistochemical features were diagnostic of ACC. ACC is often presented as a gingival lesion; thus, it may easily be neglected by patients. The identification of this tumor using specific pathological analyses prevents misdiagnosis and enables clinicians to determine the appropriate treatment. In this case, no recurrence or distant metastasis was observed after 2 years of follow-up.

Using poly(lactic-co-glycolic acid) microspheres to encapsulate plasmid of bone morphogenetic protein 2/polyethylenimine nanoparticles to promote bone formation in vitro and in vivo.

Repair of large bone defects is a major challenge, requiring sustained stimulation to continually promote bone formation locally. Bone morphogenetic protein 2 (BMP-2) plays an important role in bone development. In an attempt to overcome this difficulty of bone repair, we created a delivery system to slowly release human BMP-2 cDNA plasmid locally, efficiently transfecting local target cells and secreting functional human BMP-2 protein. For transfection, we used polyethylenimine (PEI) to create pBMP-2/PEI nanoparticles, and to ensure slow release we used poly(lactic-co-glycolic acid) (PLGA) to create microsphere encapsulated pBMP-2/PEI nanoparticles, PLGA@pBMP-2/PEI. We demonstrated that pBMP-2/PEI nanoparticles could slowly release from the PLGA@pBMP-2/PEI microspheres for a long period of time. The 3-15 µm diameter of the PLGA@pBMP-2/PEI further supported this slow release ability of the PLGA@pBMP-2/PEI. In vitro transfection assays demonstrated that pBMP-2/PEI released from PLGA@pBMP-2/PEI could efficiently transfect MC3T3-E1 cells, causing MC3T3-E1 cells to secrete human BMP-2 protein, increase calcium deposition and gene expressions of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), SP7 and I type collagen (COLL I), and finally induce MC3T3-E1 cell differentiation. Importantly, in vivo data from micro-computed tomography (micro-CT) and histological staining demonstrated that the human BMP-2 released from PLGA@pBMP-2/PEI had a long-term effect locally and efficiently promoted bone formation in the bone defect area compared to control animals. All our data suggest that our PLGA-nanoparticle delivery system efficiently and functionally delivers the human BMP-2 cDNA and has potential clinical application in the future after further modification.

[Study of 5-bromodeoxyuridine labeling of endothelial progenitor cells from the circulating blood from tooth movement rat].

OBJECTIVE: To investigate the optimal dosage and timing for 5-bromodeoxyuridine (BrdU) labeling of endothelial progenitor cells (EPCs) from rat circulating blood. METHODS: The animal model for rat tooth movement was established. EPCs were obtained by density gradient centrifugation. The expressions of specific antigens on cell surface were analysed by immunocytochemistry and fluorescenceochemistry. EPCs were incubated with BrdU at different concentrations (5, 10, 15 micromol/L) for different incubating time (12, 24, 48, 72, 96 h) to identify the optimal BrdU concentration and incubating time for cell labeling. Immunohistochemistry was performed to calculate the labeling index (LI). RESULTS: The culture cell positively expressed CD34, CD133 and could be shown to endocytose DiI-ac-LDL, FITC-UEA-1. Incubation of the EPCs with BrdU at 10 micromol/L and for an optimal length of 72 h appeared to achieve the highest LI (66.8+/-2.9)%, which was significantly higher than group of 5 micromol/L (P<0.05), while there was no significant difference between the group of 15 micromol/L and 10 micromol/L (P>0.05). CONCLUSION: EPCs can be isolated from tooth movement rat circulating blood and cultured. Incubation of the EPCs with BrdU at 10 micromol/L and for an optimal length of 72 h appeared to achieve the optimal LI. This provides a foundation for us to investigate the mechanism of chemiotaxis and differentiation for EPCs.