Objective：To investigate the mechanism of ClC-3 chloride channel in the progress of PTH regulating bone osteogenic differentiation. Methods：Regulate the expression of TGF-β1 and ClC-3 chloride channel in MC3T3-E1 cells with specific siRNA. Investigate the expression of Clcn3、Tgfb1 and osteogenic factors (Alp, Bsp, Oc and Runx2) with the method of Real-time PCR. Investigate the expression of CLC-3 chloride channel protein and TGF-β1 protein with the method of Western Blot and immunofluorescence. Results: The effect of TGF-β1 gene expression on ClC-3 expression in osteoblasts under discontinuous stimulation of PTH: ClC-3 chloride channel gene and protein expression increased when TGF-β1 gene expression in osteoblasts was interrupted only. The effect of ClC-3 chloride channel gene expression on TGF-β1 expression in osteoblasts under discontinuous stimulation of PTH: TGF-β1 gene and protein expression increased when ClC-3 chloride channel gene expression was interrupted only. Expression of osteogenic genes under discontinuous stimulation of PTH: Expression of osteogenic genes (Alp, Bsp, Oc and Runx2) decreased comparing to control when ClC-3 gene and TGF-β1 gene were interrupted respectively (P<0.05). However, no significant difference was observed between the expression of the simultaneously interrupted genes and the control （P>0.05）. Conclusion：ClC-3 chloride channel has been involved in the osteoinduction of PTH in the osteoblasts through the TGF-β pathway. The specific mechanism behind that requires further research.

[Abstract] Objective：To observe the effect of gambogic acid (GA) derivative compound 5 (C5) on apoptosis in human head and neck squamous cell carcinoma (HNSCC) in vitro. Methods：In the present study, HNSCC cell lines CAL27, HN13 and HN30 were treated with C5 at different doses for different times. The cell viability was detected with MTT assay and the IC50 values were also determined by Logit method. Cell apoptotic rates induced by C5 in the three cell lines were observed by Annexin-V/PI doubling staining by the flow cytometry assay. The changes on the s of apoptosis-related proteins were detected by Western blot and fluorescent immunocytochemistry assay. Results：CAL27, HN13 and HN30 cell lines were dramatically inhibited in a dose- and time-dependent manner after incubation with C5. The IC50 values were less than GA. When CAL27 cells were incubated with high dose C5 of 5.0 μmol/L for 24h, 48h and 72 h, the apoptotic rates were about 52.19%, 65.24% and 84.53%, respectively. C5 decreased protein levels of Bcl-2 and increased protein levels of Bax in the CAL27, HN13 and HN30 cell lines. Conclusions：Altogether, our data demonstrated that C5 induced apoptosis of HNSCC cell lines with up-regulation of Bax and down-regulation of Bcl-2 in vitro.

The purpose of this study was to discuss the correlation of autophagy and angiogenic capacity in co-culture using human amniotic membrane-derived mesenchymal stem cells (hAMSCs) with human umbilical vein endothelial cells (hUVECs) in 3D gels by comparing to hUVECs-monoculture. Methods：To isolate hAMSCs and hUVECs, Amnion were incubated with dispase and collagenase, while umbilical veins were digested with collagenase type 1. Identification of hAMSCs and hUVECs through Flow Cytometry , immunofluorescence staining and Multilineage differentiation. The length of sprouting and average vessel area were surveyed in hUVEC-hAMSC co-culture and hUVECs-monoculture at 0, 3, 6, 12, 24, 48, 72h. ATG5, Beclin-1 and LC3Ⅱ was analyzed in hUVEC-hAMSC co-culture and hUVECs-monoculture by western blotting a 0, 3, 6, 12, 24, 48, 72h. Results：hAMSCs were positive for MSC markers, while negative for hematopoietic and vascular cellrelated markers. hUVECs were positive for CD34 and negative for CD45. hUVECs expressed CD31 on membrane surface and vWF in cytoplasm. The length of sprouting and average vessel area in hUVEC-hAMSC co-culture were more than those in hUVECs-monoculture after 12 h. Expression of ATG5 in hUVEC-hAMSC co-culture increased significantly in hUVECs –monoculture at 3 and 6 h, while Beclin 1 and LC3Ⅱ rose remarkably in hUVEC-hAMSC co-culture before 6 h and 12h respectively. Conclusions：In conclusion, this study discovered that hAMSCs can promote angiogenic capacity and demonstrated that the capacity of promote angiogenic had intimate connection with elevated level of intracellular autophagy at early co-culture system.

Object: To establish digital simulated teeth roots for three dimensional printed scaffolds to preserve alveolar ridge after tooth extraction, and verify its similarity with the extracted teeth and the tooth socket. Methods: The left mandibular incisor was extracted with minimally invasion. The incisor and mandibles were scanned by Micro-CT, and a digital tooth model was established by 3D-DOCTOR software. The volume of incisor teeth, digital teeth model and teeth socket were calculated. The volume of digital teeth roots and teeth sockets were calculated by “hole filling” method. The real volume of incisor teeth was explored by flotage method. Then the data were compared to find the differences. Results: Three dimensional digital models of the skull, the left mandibular incisors and the mandibular teeth after extraction were constructed by 3D-DOCTOR software. There were no significant differences between the volume of teeth roots and digital teeth roots model (P>0.05). Both the volume of the teeth roots and that of the digital teeth were smaller than the volume of the teeth sockets (P<0.05). The volume of sockets were smaller before than after extraction (P<0.05). Conclusions: Digital simulated teeth roots for three dimensional printed scaffolds were successfully established, with high similarity to teeth roots. And its application will achieve better prognosis in alveolar ridge preservation.

Objective：To develop nano-SiO2 coated orthodontic stainless steel wires and to study the surface characters and the basic physical properties of the coating. Methods: Nano silicon dioxide film was deposited on the surface of stainless steel orthodontic wire by using unbalanced magnetron sputtering method. The surface and cross section properties of the coatings were observed with scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS).We also compared the changes of friction property between the two archwires. Results: A nano silica coated wire was thus developed, and the silicon dioxide film was about 50 nm thick. The size and the distribution of silica is uniform, and the surface is bright and clean. The friction property of the coating was obviously superior to that of the stainless steel wire. Conclusions: The nano silica coating on the surface does not change the size of the wire. With good surface properties ,better friction properties and photochemical activity it is better for clinical use.

Objective: To observe histomorphology changes of condyle and differences between young and adult through mandibular advancement intermittently so as to provide the opportunity and strategy of mandibular advancement. Methods: Mandibular advancement model was established on 4 weeks old and 8 weeks old BABL/c female rats. The TMJ samples were harvested at the 14th and 28th days and stored for Trap staining and Godener trichrome staining and Micro-CT scan. Results: Compared with those at 28th day of the 8-week-old group, the at 28th day of the 4-week-old group was as follows: BV/TV and TB.Th decreased; Tb.Sp increased(P<0.05); TB.N was not different(P>0.05). Compared with those at 14th day of the 8-week-old group, the at 14th day of the 4-week-old group was as follows: the number of Trap positive osteoclasts and the number of osteoblasts were all increased(P<0.05). Compared with those at 28th day of the 4-week-old group, the at 14th day of the 4-week-old group was as floows: the number of Trap positive osteoclasts and the number of osteoblasts were all increased(P<0.05). Conclusion: Bone remodeling on rats is related with mandibular advancement and the has significant differences between different ages.

Objective：To observe the effects of endotoxin tolerance on the production of chemokine epithelial neutrophil-activating peptide 78 (ENA-78) and epidermal growth factor (EGF) in THP-1 cells. Methods：THP-1 cells were pretreated with 1μg/ml Porphyromonas gingivalis (P.gingivalis) lipopolysaccharides (LPS) or 1μg/ml Escherichia coli (E.coli) LPS (24h), washed and stimulated with the same LPS again (24h). Levels of ENA-78 and EGF in supernatants were detected by ELISA. Results：There were no differences in production of ENA-78 secreted by THP-1 cells stimulated with P.gingivalis LPS and culture medium (p>0.05), while the amounts of ENA-78 in the cells challenged by E.coli LPS were increased significantly compared with those without any stimulations (p<0.05). In addition, secretions of ENA-78 were decreased significantly after repeated P.gingivalis LPS or E.coli LPS stimulations compared with those following signal challenge (p<0.05). However, no significant changes of EGF levels were detected in THP-1 cells restimulated with P.gingivalis LPS or E.coli LPS compared with those challenged only once (p>0.05). Conclusions：Secretions of ENA-78 in THP-1 cells could be suppressed by endotoxin tolerance, which might have an effect on neutrophil migration.

Objective: To assess the applicability and accuracy of Demirjian, Willems, Haavikko methods in estimating the dental age of Nanjing children. Methods: A total of 450 panoramic radiographs (263 males and 187 females) of Nanjing children were collected for the study on the basis of predetermined inclusion and exclusion criteria. Dental age was calculated using Demirjian, Willems, and Haavikko methods and the difference between estimated dental age and chronological age were compared with paired t-test. Results: Demirjian method overestimated age 0.50 years(boys 0.48 and girls 0.53); Willems method overestimated 0.02 years(boys 0.09 and girls 0.07); Haavikko method underestimated 0.85 years(boys 0.87 and girls 0.82). For individual tooth using Haavikko’s method, the first molar method underestimated 0.42 years(boys 0.45 and girls 0.36)；the center incisor underestimated 0.65 years(boys 0.66 and girls 0.61). Conclusions: Willems’s method was more accurate in estimating dental age compared to other methods. It is followed by Demirjian and Haavikko method; for individual tooth using Haavikko’s method, first molar and central incisor were more accurate than the mean value of all developing teeth.

There are both epithelial and lamina propria stem cells in the oral mucosa. After analyzing and coordinating accepted literatures, we have made a review about the research progress of the separation, cultivation, identification, plasticity and application for oral mucosal stem cells. They will be a potential source of stem cells, which may contribute to cellular tissue engineering, regeneration and the therapy.

The O class of Forkhead (FoxO) transcriptional factor belongs to the Forkhead proteins family, regulating bone coupling, development of osteolytic lesions and bone homeostasis in chronic inflammations. Chronic apical periodontitis is a kind of chronic inflammatory disease occurring in the periapical tissues, leading to periapical bone loss. So insights of how FoxO transcription factors act on bone metabolism maybe contribute to seek new treatment for chronic apical periodontitis. In this review, the role of FoxO transcription factors on cell proliferation, apoptosis and osteogenetic differentiation, as well as the relationship between FoxO and inflammation, inflammation factors, oxidative stress are presented .

In recent years, dentition defect as one of the most common diseases of the dental clinic has attracted more and more attention. The tooth autotransplantation is also increasingly accepted because of the economic and artistic advantages.In order to realize the delayed tooth autotransplantation , cryopreservation technology has become a hot research topic at home and abroad.This paper mainly expounds the research background and theory of cryopreservation technology, mechanism of cryoprotectant,introduces main methods and techniques of cryopreservation, discusses the effect of cryopreservation on the biomechanical properties of human dental tissues, and outlooks the application prospect of tooth cryopreservation .

Significant methodological differences exist in the isolation and culture ofPDLSCs(PDLSCs) from rat and human. The isolation and culture of rat PDLSCs is restricted by theobstructions ofthe small donor molars, relativelimitedperiodontal ligament sources, and the difficulty of tooth extraction. However, the establishment of rat model of periodontitis is widely usedin the basic research, the isolation and culture of rat PDLSCs are of vital necessity for exploring the pathology of periodontitis. Furthermore, they are of additional importance for finding out whether the environmental changesimpair biological properties of PDLSCsto cause serious damage of periodontal tissues and the difficulties in regeneration.On the other hand, after the establishment of the methodology in the isolation and culture of rat PDLSCs, theoretical basis of stem cell-based periodontitis treatment and periodontal regeneration could be provided. Therefore, it is vitally significant to summarize the isolation and culture of rat PDLSCs that have promising applicable prospects.