Objective Mixed culture of RFP markers of tooth germ cells and bFGF/BMP2 transfecting the bone marrow stem cells(BMSCs ) was conducted and polyvinyl alcohol (PVA)/biphasic ceramic bone scaffold (DCCP) stand was painted threedimensionally to construct heterogeneous tooth tissue engineering(HTTE). In order to investigate their ability to differentiate in vivo. Methods 128 healthy SD rats were randomly divided into four groups: the cell pellet group; PVA/DCCP scaffold group; cell clumps + PVA/DCCP scaffold group; control group which was without any intervention. The HTTE was implanted into the renal subcapsular of rats under general anesthesia. Four time points after surgery were chosen to collect samples for testing, Immunohistochemical staining and confocal slice observation. At the 5th, 10th, 14th, 28th day, 8 samples were collected from each group. Results Immunohistochemistry: Factorial analysis showed that cell clumps + PVA/DCCP scaffold group in 5, 10 days DMP1, BMP4 expression were the highest and were higher than the rest groups, the expression level gradually decreased over time, and the difference was statistically significant P<0.05; DSP in each group at each time point was negative, the difference was not statistically significant P>0.05. Confocal microscopy showed: cell clumps +PVA/DCCP scaffolds group at each time point biopsy tissue could be observed in tooth germ cell fluorescently marker. Conclusion RFP labeled tooth germ cells and bFGF/BMP2 transfecting BMSCs mixed culture, composite PVA/DCCP scaffold successfully construct tooth tissue engineering and tooth differentiation, but its differentiation mechanism needs further study.

Objective To explore the characteristic of different cartilage staining methods for condylar fibrocartilage and femoral hyaline cartilage in rabbit. Methods Temporomandibular joints and knee joints were harvested from healthy female Newzealand White rabbits. HE staining, Toluidine blue staining, Alcian blue staining, Safranin OAlcian blue staining, Safranin O staining, Safranin OFast green staining were used to observe and analyse the organizational structure of the cartilage under optical microscope. Results Organizational structure of the articular cartilage was clear in HE staining, such as cartilage matrix was dyed blue and bone was dyed red. Articular cartilage matrix was dyed violet and bone was not stained in Toluidine blue staining. Articular cartilage matrix was dyed light blue and bone was not stained in Alcian blue staining. Articular cartilage matrix was dyed light blue and bone was dyed light red in Safranin OAlcian blue staining, but the fiber layer and proliferation layer of condylar cartilage were dyed red. In Safranin O staining, bone was dyed light red, whereas the cartilage matrix of the condyle was dyed red and the cartilage matrix of the femur was dyed orange. Articular cartilage matrix was dyed red and bone was dyed green in Safranin OFast green staining. Meanwhile, the fiber layer of the condylar cartilage was dyed green. Conclusions Different cartilage staining methods show structure of cartilage tissue specificity. We recommend that we should choose suitable cartilage staining methods according to our research purpose and specimen in the future study.

Objective Fusobacterium nucleatum (F. nucleatum), Gramnegative bacteria, is one of the important pathogens of periodontal disease. The studies have found that periodontal pathogen infection and oral cancers development are closely related. Therefore, this study aims to analyze the effects of bacterial infection on cell proliferation and cytokine secretion, providing new evidence for the relationship between F. nucleatum infection and oral cancer development. Methods KB cells infected with F. nucleatum model was established. The morphology of bacteria and cell was observed by transmission electron microscopy. Cell cycle was analyzed by PI staining flow cytometry, the levels of IL6 and IL8 cytokines in the supernatant were detected by ELISA. Results F. nucleatum invaded the cells at 4 h. The bacteria remained intact within the cells, and the tight connection was broken among cells. 24 h after infection, F. nucleatum could still be found within the cells. The bacteria capsule structure wrapped around and maintained integrity. Low concentration of F. nucleatum (MOI=10) infected KB cells promoted G1 phase process at 4 h(P<0.05), but had no significant changes in cell proliferation. High concentration of F. nucleatum (MOI=100) infected KB cells promoted cell proliferation through accelerating S phase cells at 4 h8 h. However, after 24 h infection, the G1 phase was inhibited and the S phase was significantly reduced resulting in cell proliferation blocked (P<0.05). The levels of IL6 and IL8 cytokines in the supernatant increased and the time and concentration were dependent on the KB cells with F. nucleatum. Conclusions Infection of oral cancer cells with F. nucleatum may aggravate the malignant course of cancer cells by affecting cell proliferation and cell cycle progression, exacerbating cell inflammatory responses.

Objective To analyze the application of the novel chitosangelatin 3D scaffold in socket preservation therapy. Methods The novel chitosangelatin 3D scaffold was composed by dimethyl sulfoxide (DMSO)/UV crosslink. Then the scaffold was compounded with rhBMP2. Eight beagle dogs, sixteen sockets after teethextraction in the left and right mandibles were selected. The sockets were divide into control group, empty scaffold group (S0 group), scaffold+ heparin group (SH group), scaffold+BMP2 group (SB group) by random. The scaffolds were implanted in each group following the group division. The animals were put into death 90 days after the surgery. The heights and widths of each socket’s alveolar bone were measured. Then the socket’s alveolar bones were analyzed by micro CT and histology. Results The alveolar bone width and height loss in S0, SH, SB groups were less than those of control group (P<0.05). By comparison between groups, SB group and SH group were less than S0 (P<0.05). Micro CT 3D reconstruction showed that S0 group, SH group and SB group were better in retaining the original alveolar ridge width and shape. The control group obviously had narrow alveolar ridge, and there were some local absorptions; histologically, in SB group no bone was absorbed, a mature bone trabeculae formed and bone calcification degree was higher than other groups. Conclusion The new threedimensional scaffold can achieve good bone formation in animal models, which can maintain the height and width of alveolar ridge after tooth extraction.

Objective To investigate the inhibitory effect of HCPT/PEGPLA nano micelles on RSCC1 cells in vitro. Methods Rabbit tongue cancer cells RSCC1 were recovered, cultured and passed to the logarithmic growth phase experiment. After cocultured for different times with PBS group, different concentrations of HCPT group and HCPT/PEGPLA group, MTT method was used to detect cytotoxic effect in vitro, and cell growth inhibition rate was calculated. Results MTT assay showed that HCPT common dosage group showed significant cytotoxicity and the growth inhibition rate increased with the increase of drug concentration and action time. The growth inhibition rate of HCPT/PEGPLA nano micelles dosage group at 48h was less than HCPT common dosage group (P<0.05), and at 96 h,there was no statistical difference compared with HCPT common dosage (P>0.05). Conclusions ① HCPT/PEGPLA nano micelle has a direct inhibitory effect on the RSCC 1 cells, showing time and dosedependent properties. ② HCPT/PEGPLA nano micelle has sustained release.

Objective To compare the apical and coronal sealing ability of three kinds of root canal filling materials. Methods 80 single rooted human anterior teeth were prepared and divided into four experimental groups(n=20) to evaluate the leakage 7 days and 90 days later. Group A and C used lateral compaction guttapercha with AHplus. Group B and D were filled with GuttaFlow+single cone. Results The apical leakages of four groups were (0.67±0.67), (0.51±0.76), (3.07±1.09), (2.10±0.59)mm respectively,the coronal leakages of four groups were (0.42±0.46), (0.53±0.46), (2.62±0.98), (2.41±0.68)mm respectively. 〖JP2〗Significant differences were found between group A and group B, group C and group D in both apical and coronal leakage length. Conclusion The apical and coronal sealing abilities of GuttaFlow+single cone technique are greater than AHplus, so GattaFlow system can be applied in clinic.

Objective To observe the relationship between Notoginsenoside R1(NGR1) and osteoblasts. Methods The osteoblastic progenitor cells MC3T3E1 were cultivated in alpha MEM culture medium containing 10%FBS (GibcoTM Invitrogen). Different concentrations and different kinds of NGR1 were added for searching the effect of NGR1 on osteoblasts in vitro. The following were measured in this experiment: quantitative fluorescence methods were used to detect cell proliferation; cell osteogenic differentiation alkaline phosphatase (ALP) activity was detected by colorimetric; OCN content in the medium was assayed through enzymelinked immunosorbent assay (ELISA). The expression level of each gene was calibrated through realtime fluorescence quantitative RTPCR detection, and the expression level of housekeeping beta actin gene was taken as an internal reference. Results When the concentration of NGR1 was 50 mg/L,it could significantly promote cell proliferation, but the effect of increasing concentration decreased, more than 100 had the function of inhibiting cell proliferation; when the concentration of NGR1 was 50 mg/L, it could significantly promote the activity of ALP, with the increasing of NGR1 concentration, the activity of ALP increased, and reached a plateau after the fall. NGR1 upregulated early, the expression of OCN was not significantly different, the expression of OCN increased when the concentration of NGR1 (5200 mg/L) was upregulated after 7 days. There was no significant difference in the overall expression on the 7th day, however on the 7th day at the concentration of 200 mg/L the most significant effect was induced; NGR1 significantly promoted the proliferation of osteoblasts, and gradually increased with the increasing of the concentration of NGR1. Conclusion NGR1 at fixed concentrations can promote osteoblasts proliferation.

Objective To examine the function of KLF6 in the oral lichen planus pathogenesis and canceration. Methods Immunohistochemistry and Westernblot were used to examine KLF6 protein in 10 cases of normal oral mucosa, 30 cases with oral lichen planus and 22 cases with squamous cell carcinoma. SPSS 14.0 software package was used for t test. Results The expression level of KLF6 protein in 30 cases with oral lichen planus was lower than that in 10 cases with normal oral mucosa and there was significant difference between the two groups(P<0.05).The expression level of KLF6 protein in 22 cases with squamous cell carcinoma was lower than that in 30 cases with oral lichen planus.(P<0.05) and the two groups had significant difference. Conclusions The abnormal expression of KLF6 may play a role in the development and carcinogenesis of OLP.

Objective To determine whether under the Emax ceramic block with different thicknesses and different colors the light cured resin can be fully cured, to facilitate clinical Emax ceramic veneer aesthetic restoration of microdontia, teeth when reversed, and to set the maximum thickness of the stick side. Methods By using Fourier transform infrared spectroscopy,the curing rate of light composite resin of EMAX porcelain block was measured by CEREC chip with different colors (A1, A4, B1, B4, C1, C4) and different thicknesses (2.0 mm, 2.2 mm, 2.4 mm, 2.6 mm, 2.8 mm, 3.0 mm) . Results When the illumination time was 120 s, and A1, B1, C1 color ceramic thickness was less than or equal to 2.6mm, the resin was fully cured. When A4, B4, C4 color ceramic thickness was less than or equal to 2.2 mm, the resin was fully cured. Curing rate was not statistically different between the light colored ceramics（P＞0.05）. There was a statistical difference in the cure rate between the light colored ceramic and the dark ceramic（P＜0.01）. The cure rate of the resin of dark ceramic was obviously lower than that of the light colored ceramic. Conclusion The curing rate of the resin obviously decreases with the increase of the ceramic color and the thickness, the thickness and color of Emax ceramic have obvious influence on the cure rate of light cured resin.

Objective To retrospectively analyze risk factors which influence early implant failure under a large sample data. Methods A total of 8 852 patients with 15 204 dental implants were collected from The Affiliated Stomatology Hospital, School of Medicine, Zhejiang University during the years of 2011-2015, which included 4 413 males and 4 439 females, 11 899 Straumann group implants, 1 993 Nobel group implants and 1 312 Ankylos group implants. Age, gender, location, implant brands, implant size and bone augmentation procedures were recorded. Chi square test was used to identify risk factors associated with early implant failure. Results Ninetyseven implants in 83 patients failed before or at the provisional restorations, and the 5year early implant failure rate was 0.64%. After 2013, the early failure rate of implants decreased year by year, and there was no significant difference between the early failure rates of implant systems. The early failure rate of male patients was significantly higher than that of female patients, and the failure rate of patients over 61 was significantly higher than that of patients within 41~60 and below 40 . The number of implants in mandibular anterior area was the smallest, while the failure rate of implant was significantly higher than that in other areas. The failure rate of narrowdiameter implant (≤3.5 mm) was higher than that of regulardiameter and widediameter implant, while the early failure rate of long implant (>13 mm) was significantly lower than the other two lengths of implants. Conclusions Elderly male patients are more likely to develop early implant failure. The early failure rates of narrowdiameter implants, short implants and implants at the mandibular anterior are much higher.

Objective To compare the ability of singlevisit RCT and 2visit RCT to remove bacteria in infected root canal. Methods There were 100 patients with chronic periapical periodontitis who were randomly divided into singlevisit group and 2visit group. Bacteria culture was taken before and after the root canal preparation(or after disinfection). The change of colony number before and after the preparation of two groups was compared. The sample after preparation(or after disinfection) was taken, DNA was extracted from bacteria, PCR was amplified, and the sequence was detected to identify bacterial species. Results Comparing the amount of bacteria of singlevisit group(after preparation)with 2visit group(after disinfection), the difference was not statistically significant. There was no significant difference between the two groups in Streptococcus oralis, Lactobacillus acidophilus, Enterococcus faecalis, Actinomyces naeslundii, Peptostreptococcus anaerobius, and Prevotella intermedia (P>0.05). However, the detection rates of the Fusobacterium nucleatum and Porphyromonas gingivalis in 2visit groups were higher than those in singlevisit group, and the difference was statistically significant (P<0.05). Conclusion 2visit RCT cannot reduce the number and species of bacteria significantly. There is no significant difference between two methods.

Objective To evaluate the correlations between XIAP expression and oral squamous cell carcinoma (OSCC) metastasis, and to make a preliminary evaluation on whether the expression can be seen as the new target gene of prognosing the OSCC. Methods 40 cases of primary OSCC and 11 cases of postoperative recurrence were collected. The expression of XIAP was examined by IHC and its correlation with patients’ clinical parameters was evaluated. Results Totally 40 cases were investigated and XIAP was expressed in varying degrees in the cytoplasm of cancer cell. There was a significant correlation between cytoplasm XIAP expression and the Lymph node metastasis, pathological stage and recurrence. The expression of XIAP in recurrent patients was significantly higher than primary tumors. Conclusion XIAP expression level is closely related to the local lymph node metastasis of oral squamous cell carcinomas and postoperative recurrence, which may be the new gene to promote the lymph node metastasis of OSCC.

Objective To study comparatively the impact of invisalign and fixed appliance on the periodontal health of the orthodontic patients through their changes of the periodontal indexes in the orthodontic process. Methods 20 adult patients who were treated with fixed and invisible orthodontic treatment from March 2013 to April 2015 were selected from the Affiliated Hospital of Wenzhou Medical University. Patients periodontal indexes(including GI, SBI, PLI, SPD) were examined respectively before the treatment, one month after accepting treatment, six months after accepting treatment, when completing the treatment, and three months after the treatment. Results At two time points, namely before the treatment and three months after the treatment, there was no obvious difference in the periodontal indexes between the two patient groups. During the whole process of the orthodontic treatment, the periodontal indexes of the invisalign group had a downward trend whereas those of the fixed appliance group had increased significantly. Besides, the sulcus probing depth did not differ between the two groups. Conclusion The use of traditional fixed appliance might cause deterioration of oral conditions, while the invisaligh can well protect the periodontal tissues.

Objective To evaluate the clinical application of lithium disilicate glassceramic in the treatment of small anterior teeth restoration. Methods 87 teeth of 45 patients were restored by IPS e.max press ceramic veneers. The teeth underwent a detailed clinical examination according to the modified California Dental Association (CDA)/Ryge standards including marginal fitness, ceramic surface, color match, sensitivity, gingival health after 1 month, 6 months, 12 months, 24 months and 36 months. Results 11 teeth showed the sensitive symptoms at the review after 1 month and the symptoms were significantly improved after treatment. After 6 months, 9 teeth showed the inflammation of gums and the symptoms were improved after health education and periodontal treatment. Review after 1 month and 6 months showed that all the veneers had good marginal fitness, integrity and ceramic surface. Review after 12 months showed that a veneer fell off, and was reattached to the tooth surface after cleansing. Review after 24 months showed that a veneer collapsed during eating, and the dentin was exposed. The veneer was remanufactured and restored. Only 7 teeth showed obvious gingival retraction at the review after 36 months. Conclusion Lithium disilicate glassceramic veneer is suitable for the treatment of anterior teeth and can achieve good clinical effect and longterm prognosis.

Objective To explore the clinical characteristics of the fourth molars. Method The panoramic radiographs of the patients from department of stomatology, Changsha Central Hospital, from June 30,2013 to June 30,2016, were reviewed. The patients who got supernumerary teeth at the distal of the third molar were selected, then their names, sexuality, age, attendance reason, the number of supernumerary teeth, their position, pattern and whether they were erupted, were recorded and analyzed. Result Twentytwo fourth molars in 21 cases were found, and the sexual ratio was 8∶13. Eight fourth molars of the twentytwo were at the left maxillary, eleven were at the right maxillary, three were at the left mandible and no one was at the right mandible. Five of them were erupted, and the rest were still impacted in the bone. By observing the pattern of tooth, ten teeth were conic, six were nodular, four were supplementary type and two of them were the fused teeth. The disparity between fourth molar at maxillary and mandible was remarkably divergent(P<0.01);the disparity whether the fourth molar was erupted or not was remarkably divergent (P<0.05). Conclusion The fourth molar at the maxillary is more common, and a majority of them are still embedded in the bone. We advocate the patients who have fourth molar which is embedded in the bone and dont have subjective symptoms should be observed closely.

Objective To evaluate the postoperative pain and the longterm clinical effect in treating the chronic apical periodontitis of the maxilla or the mandibule with the rotary NiTi instruments and the ultrasonic root canal irrigation which were applied jointly，independently or neither. Methods A total of 280 subjects with chronic apical periodontitis of maxilla and mandibule were selected and categorized into 4 groups randomly. The control group(n=69) was prepared by kfile and irrigated by injector. The NiTi group(n=70) was prepared by NiTi instruments and irrigated by injector. The ultrasonic irrigation group(n=70) was prepared by kfile and irrigated by ultraphonic. The union group(n=71) was prepared by NiTi instruments and irrigated by ultraphonic. All the groups were filled at once, and the postoperative pain reaction was evaluated after a week; after 2 years, the longterm clinical efficacy was assessed. Results After a week, the postoperative pain reaction of the control group was the most serious, whose incidence rate was 50.7%. The NiTi group and the ultrasonic irrigation group were slightly better, whose incidence rates were respectively 28.6% and 27.1%. The union group had the best result, the incidence rate of which was only 9.86%. After 2 years, there was no significant difference in the longterm clinical efficacy between any two groups（P>0.05）,besides the statistically significant difference between the control group and the union group（P<0.02）. Conclusion The treatment of the chronic apical periodontitis of the maxilla or the mandibule with the rotary NiTi instruments and the ultrasonic root canal irrigation jointly can reduce the incidence rate of the postoperative pain reaction, and can improve the longterm clinical efficacy, so it is wroth being promoted.

Objective To assess the influence of mothers’ different sociodemographic backgrounds, their own oral health habits, feeding habits and oral health status on their onemonth infants’ risk of dental caries. Methods Onemonth infants and their mothers from Huangpu district in Shanghai were involved in the study. Information was collected using questionnaires. Oral clinical examinations were performed and Plaque samples of mothers and their infants were also collected to assess their level of risk on dental caries. Results A total of 165 onemonth infants and their mothers participated in this study. 10.3% of onemonth infants and 41.9% of mothers were in the high risk of dental caries. DMFS of mothers ranged from 0 to 26 with a mean of 2.7±3.9. Mean values of VPI and BOP were 0.42±0.19 and 0.14±0.15.The result of the multivariable Logistic analyses indicated that the infants who were born underweight (OR=6.5, 95% CI=1.5-29.3), fed with nursing bottle more than 5 times every day (OR=5.1，95% CI=1.3-19.5), and their mothers who had more than 25% of site with bleeding (OR=4.7, 95% CI=1.3-16.8) were in the high risk of dental caries. No other sociodemographic background, oral health habits, and feeding habits were significantly associated with onemonth infants’risk of dental caries (P＞0.05). Conclusions A number of onemonth infants have high risk of dental caries. Underweight when they were born, frequency of feeding with nursing bottles and mothers with serious gingivitis are associated with onemonth infants’ risk of caries.

In order to improve the implant osseointegration and increase success rates of implant treatments, cell sheet technology provides a new method for surface modifications of implants from the perspective of biology. As a cell processing method capable of retaining extracellular matrix, with no scaffolds, the cell sheet can be combined with conventional implant materials to form cell sheetimplant complexes. Moreover, more and more studies have shown that the cell sheet can be used as a genetic engineering carrier to promote implant osseointegration through carrying with related osteogenic genes. However, this technique also has some problems in the application of implants, such as single fabrication method of the complexes and the unknown viability of the complexes in vivo. Therefore, this paper summarized the fabrication methods of the cell sheet and cell sheetimplant complexes, and tracking approaches of the cell sheet in vivo.

　 Implant failure is usually caused by periimplantitis. The abutment connection mode, surface morphology, microtopography and chemical composition play important roles in the onset and severity of periimplantitis. This review focuses on how to prevent the periimplantitis by optimizing the cervical design of dental implant.

The etiology of temporomandibular disorders has not been fully elucidated, whose causes are mainly from many aspects such as occlusal factors, sociopsychosocial factors, anatomical factors, immune factors and joint overload. The occlusal factors, as a kind of factors affecting the temporomandibular joint disorders, play a pushing role in the occurrence and development of temporomandibular joint disorders. The curve of occlusion, as the representative, plays an important role in the pathogenesis of TMD. The occlusion, temporomandibular joint and masticatory muscles constitute the core of oral masticatory system, which interrelate with and influence each other.

The oral cavity is an environment for the colonization of up to 700 known bacteria. In recent years, it has been found that the effect of artificial implantation of probiotics on oral pathogens has been paid more and more attention by domestic and foreign scholars. Oral probiotics have shown the effects on prevention and treatment of dental caries, gingivitis and periodontitis by inhibiting oral pathogen, and the literature has also verified their effect on prevention and treatment of peri implantitis. This paper reviewed the prospect of the types of oral pathogenic bacteria and probiotics, the influence of oral probiotics and the mechanism of oral probiotics.

Astragalus membranaceus is one of the common traditional Chinese medicine and the medical community is gradually doing indepth research on its pharmacological effects. Astragalus membranaceus not only has the effects of regulating immunity and blood glucose, treating cardiovascular disease and urinary system disease, antitumor, antibacterial and antiviral, but also can inhibit the periodontal pathogens, treat diseases of the oral mucosa and promote periodontal tissue regeneration, and so on. This review describes the advances of the application of astragalus membranaceus in oral medicine.