BACKGROUND: Distractor has decided the development of distraction osteogenesis since it was applied in oral and maxillofacial surgery. How to breach the limit of traditional distraction osteogenesis lies on designing three-dimensional (3-D) distractor.OBJECTIVE: To explore the design and manufacture of 3-D external distractor for zygomatic bone.SETTING: Department of Oral and Maxillofacial Surgery, First Affiliated Hospital of Anhui Medical University.DESIGN: An experiment of the external distraction system for 3-D distraction osteogenesis.MATERIALS: The distractor was made of biological material titanium with well histocompatibility, and consisted of three parts (support, expansion and direction-change) as well as some fittings such as titanium backup plate, spanner and rubber mat, etc.METHODS: Caprine of 10 months old were selected to separate the heads and prepare for isolated zygomatic models. Zygomatic bones were generally curve and made up of four-process-in-one, with the identical formation as human being. According to principle of mechanical movement,two support plates were designed to move relatively, which transferred randomly along with two perpendicular directions. It was done to change the direction and model 3-D distraction osteogenesis on zygomatic bone of the isolated caprine cranium.MAIN OUTCOME MEASURES: 3-D transference of expansion board and whole stabilization of distractor were observed in the process of distraction osteogenesis.RESULTS: The self-prepared 3-D external distractor was 30 g. The lengthener was built to provide 3-D suture expansion osteogenesis by changing the direction. The expansion bar could be moved about 2 cm in anteroposterior axes, 3.5 cm in perpendicular axes, 3 cm in coronal axes.CONCLUSION: The 3-D distractor is simple, accurate and practicable in experimental study, and lays a foundation for clinical study in the future.

Objective:To study the anatomic basis of superior trapezius myocutaneous and spina scapulae osteomyocutaneous flaps pedicled with transverse cervical vessel.Methods:The blood vessels,the size of the superior trapezius muscle and the spina scapulae were dissected and examined in 32 adult corpses.Results:The superior trapezius muscle was in the shape of trapezium.The border length (mm) of A,B,C and D was 174.63,157.18,86.98 and 80.95 in average respectively.The area of the muscle was 126.78 cm2 on the average.The spina scapulae was 131.21 mm in average length.The length (mm) of transverse cervical artery trunk, superficial cervical artery trunk and its ascending artery,spina scapulae branch artery was 42.50,27.80,43.12,28.75 in average,their external diameter(mm) was 2.71,2.39,1.96 and 0.50 respectively.Entering the muscle,the ascending artery had 3～6 branches with the external diameter of 0.5 mm or more.The venous vessels were following the copartner artery.Conclusion:The superior trapezius muscle and the spina scapulae may be made for the myocutaneous and osteomyocutaneous flaps pedicled with transverse cervical artery used in oral and maxillofacial reconstruction.

Objective: To investigate the effects of nerve growth factor(NGF) on the differentiation of goat bone marrow mesenchymal stem cells (BMSCs) into osteoblasts. Methods: The goat BMSCs were cultured in vitro and the marker proteins on the BMSCs surface were identified by flow cytometry. The third passage of BMSCs were randomly divided into blank control group, osteoblasts control group, NGF group and experimental group. The activities of alkaline phosphatase (ALP) and osteocalcin (0C) were examined and the of von Kossa staining method was used to observe the osteogenic differentiation. Results: CD90 and CD105 were strong positive while the CD34 and CD45 were negatively expressed in BMSCs. The activities of ALP and OC was significantly higher in the experimental group than that in the other three groups(P<0.05). The staining of von Kossa was positive and the black calcium nodules were appeared in the the osteoblasts control group. The number and the area of the calcium nodules were greater in the experimental group. But there were no significant differences of each index between the NGF group and the blank control group. Conclusion: NGF can′t induce goats BMSCs to osteoblasts, but can clearly promote the differentiation of goats BMSCs.

Objective To construct the eukaryotic expression vector of PRDX3 labeled with FLAG tag and to study its localization in human tongue cancer cell line SCC15.Methods PRDX3 gene was obtained from the breast library by PCR and cloned into PCDH vector to construct PCDH-FLAG-PRDX3.The plasmid was transiently transfected into 293T cells and the expression was detected by Western blot.Subcellular localization was detected by cellular immunofluorescence.Results The result of double digestion and sequencing showed that PCDH-FLAG-PRDX3 eukaryotic expression vector was constructed.The expression of FLAG-PRDX3 in human 293T cells was positively confirmed by Western blotting.In human tongue cancer cell line SCC15, the result of cellular immunofluorescence showed FLAG-PRDX3 was located in the cytoplasm rather than in the nucleus.Conclusion PRDX3 eukaryotic expression vector labeled with FLAG tag is constructed successfully, which is located in cytoplasm in human SCC15 cells.Construction and identification of PRDX3 could shed light on the function and mechanism of PRDX3 in tongue cancer.

Objective To understand the status quo of the illness situation and the implementation of prevention and control measures on Kashin-Beck disease( KBD ) in Shandong Province , and provide a scientific basis for assessment and acceptance of the work on eliminating KBD. Methods In accordance with the requirements of the national “Implementation Scheme for Endemic Disease Control in 2012” and the “Shandong Provincial Technical Scheme for KBD Control in 2012”. In four towns of Qingzhou, three epidemic villages were selected as the surveying villages in each town in order to understand the implementation of measures for prevention and control of KBD; information such as population of investigating villages, per capita income, farming, food and the implementation of control measures,etc were included. In each investigating village, 7 - 12 years old children were selected to receive clinical and X-ray examination. When fewer than 50 people in a investigating village , one additional endemic village was included near the investigating village , and all children aged 7 to 12 of the village were checked. The number of cases of clinical degree Ⅰ, Ⅱ, Ⅲand the detection rates, X-ray positive detection rate, the number of cases of metaphysis, epiphysis, distal end of phalanx, carpal bone, triad and the detection rates were calculated, respectively. Clinical and X-ray diagnosis were based on “The Diagnostic Criteria of Kaschin-Beck disease”(WS/T 207-2010). Results A total of 14 epidemic villages were selected to survey in four towns of Qingzhou. In the recent three years in the epidemic areas, measures of supplying Se had not been implemented;measures of resettlement and off-site education had not been implemented either. In the epidemic areas , the main crops were wheat and corn; the staple food was flour which accounted for more than 80%, and the foreign food bought accounted for only 11%. The areas of returning farmland to forest (grass) accounted for 0.32%(36/11 151) of the original arable land, and the areas of replanting economic crops accounted for 1.17%(131/11 151) of the original farmland. A total of 536 children aged 7 to 12 were examined by clinical and X-rays. Patient of KBD was not detected clinically. Six positive patients with metaphyseal changes of KBD were detected by X-rays, and the average X-ray detection rate was 1.12%(6/536). Patients with abnormal epiphysis, distal end of phalanx and carpal bone were not detected by X-rays. Conclusions KBD in Shandong Province has been effectively controlled. Since the cause of KBD is still unknown, it is necessary to build a sustainable long-term control mechanism of KBD;further improve the monitoring system and dynamic monitoring work still need to be strengthened.

Objective To construct PES1 shRNA stable expression cell lines in tongue squamous cell carcinoma ( TSCC) cells and to study the effect of knockdown of PES 1 on the growth of TSCC cells .Methods Recombinant lentivirus carrying PES1 shRNA was packaged and obtained in 293T cells.TSCC cells (Tca8113, SCC6 and SCC15) were infected with the lentivirus and selected for stable cells .PES1 expression was identified by Western blot .The effect of inhibition of PES1 on the growth and cell cycle of TSCC cells was detected by growth curve and flow cytometry .Results TSCC cells stably expressing PES1 shRNA were constructed.Knockdown of PES1 inhibited cell proliferation and induced cell cycle ar-rest at G0/G1 phase.Knockdown of PES1 inhibited expression of cyclin D1 in TSCC cells.Conclusion Inhibition of PES1 results in reduced cell proliferation , cell cycle arrest at G 0/G1 phase and reduction of cyclin D 1 expression in TSCC cells . PES1 may be a target for TSCC gene therapy .

Objective To construct eukaryotic expression vector of wild type E 4F1 and the mutant deleting amino acid region 32-81, and to detect the interaction between wild type or mutant E 4F1 and p53 and to study the effect of E4F1 on the expression level of p21.Methods Wild type and mutant sequences of E 4F1 were amplified from the mammary library using standard PCR and recombinant PCR .The sequences were cloned into pXJ 40-MYC vector to generate the MYC-E4F1 and MYC-E4F1(Δ32-81) recombinant plasmids that were transfected into 293T cells and identified by Western blotting . FLAG-p53 and MYC-E4F1 or MYC-E4F1(Δ32-81) were co-transfected into 293T cells and immunoprecipitation assay was performed to detect the interaction of wild type or mutant E 4F1 with p53.Wild type and mutant E4F1 expressing vec-tors were co-transfected into osteosarcoma U2OS cells and the expression of p21was detected.Results Recombinant plas-mids of MYC-E4F1 and MYC-E4F1(Δ32-81) were successfully constructed.Both wild type and mutant E4F1 interacted with p53.Deletion of amino acid region 32-81 of E4F1 increased the interaction .The expression level of p21 was in-creased by wild-type E4F1, but not by mutant E4F1.Conclusion The eukaryotic expression vector of wild type E4F1 and its deletion mutant is successfully constructed .Both of them interact with p53.Deletion of amino acid region 32-81 of E4F1 increases the interaction .This study contributes to further studies on the regulation and mechanism of E 4F1 on p53.

BACKGROUND: Extraction method of modified platelet-rich fibrin has been determined, and its structure and constitutes have been investigated. Animal experiments have shown that modified platelet-rich fibrin can cooperate with osteoblasts to promote osteogenesis. OBJECTIVE: To extract modified platelet-rich fibrin and to explore the effect of modified platelet-rich fibrin gel on the repair of distal bone defect of adjacent mandibular second molar after extraction of mandibular wisdom tooth. METHODS: Forty-eight patients with proximal or horizontal obstruction of unilateral mandibular third molar were selected from the Department of Stomatology of Anqing Hospital Affiliated to Anhui Medical University. Patients were randomly divided into modified platelet-rich fibrin group and control group. The third molar was removed and the tooth extraction socket was cleared by the same method. Modified platelet-rich fibrin gel made from the patient's own blood was put into the distal surface of the second molar in the modified platelet-rich fibrin group at the same time. The control group did not put any materials after tooth extraction. All patients were followed up at 1, 2, 3 and 4 months after surgery to evaluate the periodontal status of adjacent mandibular second molars, including periodontal examination and imaging analysis. Full-oral curved tomography and cone-beam CT were taken at 4 months after surgery. Statistical analysis was conducted on SPSS 21.0 software. RESULTS AND CONCLUSION: (1) There was no significant difference in the periodontal depth of the adjacent second molars between modified platelet-rich fibrin and control groups. (2) At 4 months after surgery, the bone mineral density and bone yield in the modified platelet-rich fibrin group were significantly higher than those in the control group (both P ＜ 0.05). (3) In summary, modified platelet-rich fibrin can promote osteogenesis of mandibular proximal second molar after mandibular wisdom tooth extraction.