The human embryonic stem cells (hESCs) serve as a self-renewable, genetically-healthy, pluripotent and single source of all body cells, tissues and organs. Therefore, it is considered as the good standard for all human stem cells by US, Europe and international authorities. In this study, the standard and healthy human mesenchymal progenitors, ligament tissues, cardiomyocytes, keratinocytes, primary neurons, fibroblasts, and salivary serous cells were differentiated from hESCs. The human cellular health-safety of NaF, retinoic acid, 5-fluorouracil, dexamethasone, penicillin G, adriamycin, lead acetate PbAc, bisphenol A-biglycidyl methacrylate (Bis-GMA) were evaluated selectively on the standar-dized platforms of hESCs, hESCs-derived cardiomyocytes, keratinocytes, primary neurons, and fibroblasts. The evaluations were compared with those on the currently most adopted cellular platforms. Parti-cularly, the sensitivity difference of PM2.5 toxicity on standardized and healthy hESCs derived fibroblasts, currently adopted immortalized human bronchial epithelial cells Beas-2B and human umbilical vein endothelial cells (HUVECs) were evaluated. The results showed that the standardized hESCs cellular platforms provided more sensitivity and accuracy for human cellular health-safety evaluation.

Tongue squamous cell carcinoma (TSCC) is the most common type of oral cancer and is well known for its high rate of proliferation and lymph nodal metastasis. Exploring the underlying pathways regulating TSCC could provide novel ideas for diagnosis and prognosis of TSCC patients, as well as molecular targets for treatment of TSCC. MicroRNAs (miRNAs) are small noncoding RNAs that inhibit gene expression through the 3′ untranslated regions (3′UTRs) of their target messenger RNAs. They play crucial roles in numerous biological processes, including cancer progression. Although great efforts have been made, what role miRNAs may play in the early detection and diagnosis of TSCC is not fully understood. Recently, our team has performed a series of basic and clinical researches in an attempt to investigate the relationships between miRNA expressions and prognosis of patients with TSCC and the mechanisms under regulation of TSCC. The results showed that miR-195, miR-34a, miR-29b, miR-375 and miR-26a could inhibit TSCC cells progression and development via a sophisticated network of genes. Specifically, the anti-tumor effects of miR-195 in TSCC may be partially mediated by its inhibition of CyclinD1 and Bcl-2 expression. The expression of miR-34a could inhibit migration and invasion of TSCC cell lines via targeting MMP9 and MMP14. The function of miR-29b may be through the miR-29b/Sp1/PTEN/AKT axis. Overexpression of miR-375 inhibited Sp1 expression by targeting the 3′ untranslated region of the Sp1 transcript. MEG3 and miR-26a inhibited TSCC cell proliferation, cycle progression and promoted cell apoptosis and miR-26a could increase the MEG3 expression through reduction of the expression of DNMT3B in TSCC. In light of the role of those miRNAs in diagnosis and prognosis of TSCC, we reported that decreased miR-195 and miR-375 expression was associated with poor overall survival rate of the TSCC patients, while miR-34a expression was negatively correlated with cervical lymph node metastases. Furthermore, combined low expression levels of miR-26a and MEG3 emerged as an independent prognostic factor for poor clinical outcomes in TSCC patients, suggesting that combined miR26a and MEG3 expression might prove useful as an independent biomarker of clinical prognosis among TSCC patients.

Objective：To investigate the potential association between FADS1 rs174537 polymorphism and serum proteins in patients with aggressive periodontitis， which may provide benefits for diagnosis and treatment of aggressive periodontitis. Methods: A total of 353 patients with aggressive periodontitis (group AgP) and 125 matched controls (group HP) were recruited in the study．Genotyping of FADS1 rs174537 and serum biochemical indexes were tested at the study’s start. The relationships between the levels of TP, GLB, ALB, A/G and genotyping were analyzed. Results: （1）The detection rate of allele G in group AgP was higher than that in group HP（68.1% vs. 61.2%, P=0.046，OR=1.35，95% CI 1.00－1.83）; the detection rate of genotype GG in group AgP was higher than in group HP（45.5% vs. 34.4%，P=0.029, OR=1.60, 95% CI 1.05－2.44）. (2) In group AgP, the patients with GG genotype exhibited significantly lower TP, GLB than the patients with GT+TT genotype ［(77.08±7.88) g/L vs. (79.00±4.66) g/L， P=0.007；(28.17±7.63) g/L vs.(29.88±3.49) g/L，P=0.007） and the higher A/G（1.72±0.22 vs.1.67±0.22, P=0.040）, but there was no significant difference in ALB between the patients with GG genotype and the patients with GT+TT genotype. In group HP, there were no significant differences in TP, GLB, A/G and ALB between individuals with genotype GT+TT and with genotype GG. (3)Compared with individuals with genotype GT+TT in group HP, the AgP patients with genotype GT+TT exhibited significantly higher TP, GLB ［(79.00±4.66) g/L vs. (75.20±4.53) g/L, P<0.01; (29.88±3.49) g/L vs.(26.55±2.94) g/L, P<0.01）and the lo-wer A/G（1.67±0.22 vs. 1.88±0.30, P<0.01）, but there was no significant difference in ALB. There were no significant differences in TP, GLB, A/G and ALB the between the AgP patients with ge-notype GG and the healthy subjects with the same genotype either.Conclusion: FADS1 rs174537 polymorphism is associated with aggressive periodontitis. The patients with genotype GG in group AgP had relatively lower TP，GLB and higher A/G. Genotype GG might be a risk indicator for aggressive periodontitis by reducing host defense capability and contributing to inflammatory response in the occurrence and development of aggressive periodontitis.

Objective：To determine the expression profile and potential roles of CD24 in oral squamous cell carcinoma and explore the values of CD24 function as a potential target of clinical therapy. Me-thods: Semi-quantitative immunohistochemistry was used to construct the expression profile of CD24 in 78 human oral tissues and 59 Hamster buccal pouch tissues. Real-time RT-PCR and Western blot were used to analyze the CD24 expression levels in oral DOK4 cells, oral cancer CAL-27 and WSU-HN6 cells. Then these two cancer cell lines were selected to evaluate the effect of all-trans retinoic acid (ATRA) and CD24 antibody on CD24 expression, and the proliferation and tumorsphere formation capacity of these two cell lines. Results: CD24 expression was found significantly elevated in both human and animal tissues compared with normal and benign tissues (P<0.05), as well as in oral cancer CAL-27 and WSUHN6 cells compared with DOK cells (P<0.05). CAL-27 and WSU-HN6 cells possess increased proliferative and specific tumorsphere formation capability compared with DOK cells (P<0.05). Both ATRA and CD24 antibody were able to effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells (P<0.05). Among them ATRA at least involved partially in the proliferation by down-regulating the CD24 expression (P<0.05), while CD24 antibody blocking had no effect on the CD24 expression. Conclusion: CD24 was upregulated in oral cancer and functioned as a potential factor that promoted the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells. Both ATRA and CD24 antibody might effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells and function as a potential therapy target.

Objective： To compare the effects of stromal cell-derived factor-1 (SDF-1) and granulocyte colony-stimulating factor (G-CSF) on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cell (DPSC) in vitro. Methods: DPSCs were cultured in vitro and treated with either 100 μg/L SDF-1 or 100 μg/L G-CSF. Cell counting kit-8 (CCK-8) and colony-forming unit (CFU) were used to detect the effect of SDF-1 and G-CSF on the proliferation ability of DPSC. Cell migration of DPSC was determined by wound healing assay and Transwell migration assay. The effects of SDF-1 and G-CSF on odontoblastic differentiation of DPSC were evaluated by alkaline phosphatase (ALP) staining, ALP activity and alizarin red S staining. The expression of odontoblastic-related genes such as dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP) were quantified by real-time RT-PCR. Results: SDF-1 and G-CSF promoted the proliferation of DPSC slightly, but the difference was not statistically significant. Wound healing assay showed that SDF-1 and G-CSF promoted cell migration of DPSC significantly (P<0.01), but there was no significant difference between the two factors. In Transwell migration assay, the number of migrated cells of the control group was 5.0±1.4 per sight, while the SDF-1 group was 24.3±6.8 per sight and the G-CSF group was 11.8±3.3 per sight, suggesting that cell migration of DPSC was improved significantly after being treated with SDF-1 or G-CSF, and SDF-1 was more effective than G-CSF (P<0.05). Significantly greater odontoblastic differentiation potential was found in SDF-1 group and G-CSF group based on the ALP staining. Higher ALP activity, more mineralization nodule formation and higher expressions of DMP-1 and DSPP were also found after SDF-1 or G-CSF treatment. Conclusion: SDF-1 had no significant effect on the proliferation of DPSC, but could significantly promote cell migration and odontoblastic differentiation of DPSC. Its effect on DPSC was better than G-CSF.

Objective：To investigate changes in the circum-maxillary sutures during alternate maxillary expansions and constrictions in a rat model. Methods: Twentytwo male Sprague-Dawley rats (6 weeks old) were used and divided into three groups. In maxillary expansion (ME) group (n=9), an expander was activated for 5 d, and then sacrificed. In alternate maxillary expansions and constrictions (AltMEC) group (9 animals), an alternate expansion and constriction protocol (5 d expansion and 5 d constriction for one cycle) was conducted for 2.5 cycles (25 d total), and then sacrificed. The control group comprised 4 animals with no appliances used, each of two sacrificed on day 5 and day 25, respectively. Circum-maxillary sutures (mid-palatal, maxillopalatine, premaxillary, zygomaticotemporal, and frontonasal suture) in each group were characterized histologically. Results: Histological findings of circum-maxillary sutures: in control group, the normal sutures were divided into 5 zones, the center was polygon mesenchyme zone, the lateral was mature chondrocyte zone, the next lateral was bone tissue. In ME group, the mid-palatal suture was expanded after ME, the fibre and connective tissue were extracted and invaded into cartilage zone. Osteoblast hyperplasia and new bone formation occurred. The orientation of the new bone trabecula was consistent with force. The osteoclast appeared in some area. Findings of other sutures were similar with those of control group, osteogenesis was active in some area without obvious reconstruction in Alt-MEC group, mid-palatal suture and other sutures in some area were widened after Alt-MEC with much more osteoblast hyperplasia and new bone formation. Fibre and connective tissue were extracted, distorted or even broken. Sutures in some areas were narrowed with osteoblast cytopenia and osteoclast hyperplasia. Fibre and connective tissue compressed because of the different force and orientation. Osteoclast count results showed that compared with the control group, the number of the osteoclast was increased only in the palatal suture in ME group (P<0.05). Other sutures did not show obvious changes (P>0.05). In Alt-MEC group, the numbers of the osteoclast in circum-maxillary sutures were the most, and had statistical significances (P<0.05). Conclusion: These results suggested that circum-maxillary sutures were actively reconstructed after Alt-MEC. while only midpalatal suture had active reaction after ME.

Objective：To investigate the role of bone morphogenetic protein 2/7 heterodimer (BMP-2/7) in the osteogenesis of human adipose-derined stem cells (hASCs). Methods: hASCs were exposed to three different treatments in vitro: osteogenic medium with 150 μg/L BMP-2/7 (experimental group), osteogenic medium alone (OM group) and proliferation medium (PM group). After 1, 4 and 7 days of osteogenic induction, the amount of cellular DNA was measured to investigate the cytotoxicity. After 7 and 14 days, alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP. After 21 and 28 days, the calcification deposition was determined by Alizarin Red S (ARS) staining and quantification. The expressions of the osteoblast-related genes were tested on days 1, 4, 7 and 14. In the in vivo study, 6 nude mice were used and 4 groups were set and implanted subcutaneously into the back of nude mice: (1) β-TCP scaffold only (scaffold control group); (2) β-TCP scaffold with hASCs cultured by PM in vitro for 1 week (PM control group); (3) β-TCP scaffold with hASCs cultured by OM in vitro for 1 week (OM control group); (4) β-TCP scaffold with hASCs cultured by OM with 150 μg/L BMP-2/7 in vitro for 1 week (test group). After 4 weeks of implantation, histological staining was performed to evaluate the in vivo osteogenesis of hASCs. Results: After induction for 1 day, there was no significant difference between the experimental group and the PM group on the cellular DNA content (P>0.05). After 4 days, the cellular DNA content increased under the stimulation of BMP-2/7 (P<0.05). On day 7, there was no significant difference among the three groups (P>0.05). ALP activity was higher by the induction of BMP-2/7 than in OM alone and PM (P<0.05). More mineralization deposition and more expressions of osteoblast-related genes such as Runx2, ALP, COL-1A1 and OC were determined in the experimental group at different time points (P<0.05). HE staining showed that, in the test group and OM control group, the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs, and newly-formed bone-like tissues could be found in ECM around the scaffold materials. Moreover, compared with the OM control group, more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test group. Masson’s trichrome staining showed that more expression of collagen could be observed in ECM in the test group compared with the other groups. There was small amount of expression of collagen in the OM and PM control groups. No obvious positive results were found in the scaffold group. Conclusion: BMP-2/7 heterodimer plays a significant role in the osteogenesis of hASCs and is able to enhance the osteogenic differentiation of hASCs in vitro and in vivo.

Objective：To construct human adipose-derived mesenchymal stem cells (hASCs)-biomaterial mixture 3D bio-printing body and detect its osteogenesis in vivo, and to establish a guideline of osteogenesis in vivo by use of 3D bio-printing technology preliminarily.Methods:P4 hASCs were used as seed cells, whose osteogenic potential in vitro was tested by alkaline phosphatase (ALP) staining and alizarin red staining after 14 d of osteogenic induction. The cells were added into 20 g/L sodium alginate and 80 g/L gelatin mixture (cell density was 1×106/mL), and the cell-sodium alginate-gelatin mixture was printed by Bioplotter 3D bio-printer (Envision company, Germany), in which the cells’ survival rate was detected by live-dead cell double fluorescence staining. Next, the printing body was osteogenically induced for 1 week to gain the experimental group; and the sodium alginate-gelatin mixture without cells was also printed to gain the control group. Both the experimental group and the control group were implanted into the back of the nude mice. After 6 weeks of implantation, the samples were collected, HE staining, Masson staining, immunohistochemical staining and Inveon Micro CT test were preformed to analyze their osteogenic capability. Results: The cells’ survival rate was 89%±2% after printing. Six weeks after implantation, the samples of the control group were mostly degraded, whose shape was irregular and gel-like; the samples of the experimental group kept their original size and their texture was tough. HE staining and Masson staining showed that the bone-like tissue and vessel ingrowth could be observed in the experimental group  6 weeks after implantation, immunohistochemical staining showed that the result of osteocalcin was positive, and Micro CT results showed that samples of the experimental group had a higher density and the new bone volume was 18%±1%.Conclusion:hASCs-biomaterial mixture 3D bio-printing body has capability of ectopic bone formation in nude mice, and it is feasible to apply cells-biomaterial mixture 3D bio-printing technology in the area of bone formation in vivo.

Objective：To investigate the relationship between the removal time of 0.2 mm occlusal interference and the recovery of masticatory muscle mechanical hyperalgesia in rats.Methods: Forty male Sprague-Dawley rats (200-220 g) were randomly assigned to eight groups, with five rats in each group: (1) na-ve group: these rats were anesthetized and their mouths were forced open for about 5 min (the same duration as the other groups)， but restorations were not applied; (2) sham-occlusal interference control group: bands were bonded to the right maxillary first molars which did not interfere with occlusion; (3)occlusal interference group: 0.2 mm thick crowns were bonded to the right maxillary first molars; (4) 2, 3, 4, 5, and 6 d removal of occlusal interference groups: 0.2 mm thick crowns were bonded to the right maxillary first molars and removed on days 2, 3, 4, 5, and 6. The na-ve group and sham-occlusal interference control group were control groups. The other groups were experimental groups. Bilateral masticatory muscle mechanical withdrawal thresholds were tested on pre-application days 1, 2, and 3, and on postapplication days 1, 3, 5, 7, 10, 14, 21 and 28. The rats were weighed on pre-application day 1 and on post-application days 1, 2, 3, 4, 5, 6, and 7.Results: Between the na-ve group and the sham-occlusal interference control group, there was no significant difference in the masticatory muscle mechanical withdrawal threshold of bilateral temporalis and masseters at each time point. No significant difference was detected between the contralateral side and ipsilateral side in experimental groups (P>0.05). In the 2, 3, 4, and 5 d removal of occlusal interference groups, the masticatory muscle mechanical withdrawal thresholds decreased after occlusal interference and increased after removal of the crowns and recovered to the baseline on days 7, 10, 14, and 14, respectively ［the masticatory muscle mechanical withdrawal thresholds of right masseter muscle were (137.46±2.08) g, (139.02±2.11) g, (140.40±0.98) g, (138.95±0.98) g, respectively］. In the 6 d removal of occlusal interference group, the masticatory muscle mechanical withdrawal threshold increased after removal of the crowns and became stable since day 14. There was a significant difference between the 6 d removal of occlusal interference group and the sham-occlusal interference group on day 28(P<0.05), the masticatory muscle mechanical withdrawal thresholds of right masseter muscle were (131.24±0.76) g and (141.34±1.43) g, respectively. Conclusion: After removal of the 0.2 mm thick crown within 5 days, the mechanical hyperalgesia of the rats could reverse completely. The mechanical hyperalgesia of the rats could only be relieved, but not reverse completely after removal of the 0.2 mm thick crown on day 6. As the time went on, even minor occlusal interference could cause irreversible mechanical hyperalgesia of masticatory muscles. This study suggested that occlusal interference caused by dental treatment should be eliminated as soon as possible, to avoid irreversible orofacial pain.

Objective： To investigate the antibacterial effect of different self-etching adhesive systems against Streptococcus mutans (S. mutans). Methods: Six reagents ClearfilTM SE Bond primer (SP), ClearfilTM SE Bond adhesive (SA)，ClearfilTM Protect Bond primer (PP), which contained antibacterial monomer methacryloyloxydodecylpyridinium bromide (MDPB), ClearfilTM Protect Bond adhesive (PA), positive control chlorhexidine acetate ［CHX，1% (mass fraction)］, and negative control phosphate buffer solution (PBS) were selected. They were mixed with S. mutans for 30 s respectively, then colony-forming units (CFU) were counted after incubated for 48 h on brain heart infusion (BHI) agar medium. The 6 reagents were applied to the sterile paper discs, and distributed onto the BHI agar medium with S. mutans and incubated for 24 h, then the inhibition zones were observed. CHX, PBS, PP, and SP were added on the dentin with artificial caries induced by S. mutans and kept for 30 s, then confocal laser scanning microscope (CLSM) was used to observe the live and dead bacteria after staining. The ratio of live to dead bacteria was calculated. PP+PA and SP+SA were applied on the dentin according to the manual and light cured. S. mutans were incubated on the samples for 2 h, ultrasonically treated and incubated on BHI agar medium for 48 h, then CFU was counted. The data were analyzed by non-parametric analysis and one-way ANOVA. Results:  Compared with PBS, the PP, SP, PA, SA and CHX showed the antibacterial effect on free S. mutans (P＜0.05); SP and PP showed stronger antibacterial effect than PA, SA and CHX (P＜0.05). CHX, SP and PP presented inhibition zones, while PBS, SA and PA did not. Compared with PBS, the CHX, SP and PP could lower the ratio of the live to dead bacteria significantly (P＜0.05). Cured selfetching adhesive systems did not show any antibacterial effect on the free S. mutans. Conclusion:The primer of self-etching adhesives ClearfilTM SE Bond and ClearfilTM Protect Bond showed significant antibacterial effect on free and attached S. mutans. The adhesive only showed antibacterial effect on free S. mutans before light-cured polymerization. After being cured, the self-etching adhesive systems did not show antibacterial effect anymore.

Objective：To study the effects of cells’ and bacteria’s adhesion and proliferation on different fiber diameters of polypyrrole coating with electricity.Methods: Titanium coated with polypyrrole was divided into no electrical stimulation and stimulation groups, each group had 30-60 nm, 70-100 nm, 130-170 nm diameters of the fiber. MC3T3 cells and Staphylococcus aureus (S. aureus) were inoculated on different fiber diameters of polypyrrole coating with and without electric stimulation . We gave the electrical stimulation group 100 mV for 1 h and every 24 hours gave it 1 h stimulation, and no electrical stimulation group was not managed. We used scanning electron microscope (SEM) to observe the cells’ and bacteria’s morphology. The cells were given 20 mL CCK8 solutions after 1,3,7 days’cultivation, then incubated for 2 h, the solution was transferred to 96-well plate, we measured the cells’CCK-8 of the 30-60 nm, 70-100 nm, and 130-170 nm groups by Elisa. The cells on different fiber diameters were also stained by livedead cell staining kit, TritonX-100 and DAPI. We used PBS to wash and glycerin to seal them. The live-dead situation and morphology were tested by co focal microscope. The bacterial were stained by Live/dead baclight bacterial viability kits, we detect the suspension’s D of the 30－60 nm, 70－100 nm, and 130－170 nm groups, and also observed the bacteria’s survival situation by co focal microscope.Results:  The CCK-8 of the cells with direct current stimulation was higher than that of the unpowered group （F=12.248, P=0.006）. The smaller the fiber diameter, the better was the cell’s adhesion and proliferation （F=9.261, P=0.005）. The bacterial suspension’s D of the electric group was lower than that of the unpowered group, and the fiber diameter  had no significant effect on the bacteria’s growth（F=9.641, P=0.036）. Conclusion: Polypyrrole coating with electricity can promote the cell’s proliferation and inhibit the bacteria’s proliferation, and the cell growth on small fiber diameter coating is better. There is no difference in the bacterial growth of different fiber diameter coatings.

Objective： To evaluate the effects of Er, Cr: YSGG laser on the root surface of periodontally involved teeth and healthy teeth, concerning the microstructure and the roughness. Methods: Eight freshly extracted teeth due to severe periodontitis and eight freshly extracted teeth due to orthodontic reasons or being third molar were chosen in this study. The root surface of each tooth was divided into four areas, and received four treatment methods: no treatment (control group); root planing with Gracey scaler for 30 seconds; irradiation by the lower power Er, Cr: YSGG laser; irradiation by the higher power Er, Cr: YSGG laser. Four periodontally involved teeth and four healthy teeth were used for the evaluation of microstructure using scanning electron microscope （SEM）. The other four periodontitis teeth and four healthy teeth were used for the evaluation of roughness (Ra value) using 3D profiler.Results: Smear layer was found on the teeth scaled by Gracey scaler, while the teeth irradiated by Er, Cr: YSGG laser demonstrated a melting surface with less smear layer. In the periodontitis teeth irradiated by the higher power, opening dentinal tubules could be observed. For the periodontally involved teeth, the Ra values of groups 1 to 4 were (237.4±20.0) nm, (135.7±11.9) nm (P=0.01), (463.6±49.3) nm (P<0.001) and (486.0±59.0) nm (P<0.001) respectively. For the healthy teeth, the Ra values of groups 1 to 4 were (191.4±44.5) nm, (131.6±21.5) nm (P=0.482), (463.7±34.6) nm (P<0.001) and (470.3±121.3) nm (P<0.001) respectively.Conclusion:Er, Cr: YSGG laser can affect the microstructure of the cementum of the periodontitis teeth and healthy teeth. Irradiation by the Er, Cr: YSGG laser resulted in a melting surface with less smear layer and increased the roughness in the surface of root.

Objective：To investigate the distribution and drug resistance of anaerobic bacteria in the patients with oral and maxillofacial infection. Methods: Aerobic and anaerobic bacteria cultures from 61 specimens of pus from the patients with oral and maxillofacial infection in the Department of Oral and Maxillofacial Surgery, Peking University School of Stomatology were identified. The culture type was evaluated by API 20A kit and drug resistance test was performed by Etest method. The clinical data and antibacterial agents for the treatment of the 61 cases were collected, and the final outcomes were recorded. Results:The bacteria cultures were isolated from all the specimens, with aerobic bacteria only in 6 cases (9.8%), anaerobic bacteria only in 7 cases (11.5%), and both aerobic and anaerobic bacteria in 48 cases (78.7%). There were 55 infected cases (90.2%) with anaerobic bacteria, and 81 anaerobic bacteria stains were isolated. The highest bacteria isolation rate of Gram positive anaerobic bacteria could be found in Peptostreptococcus, Bifidobacterium and Pemphigus propionibacterium. No cefoxitin, amoxicillin/carat acid resistant strain was detected in the above three Gram positive anaerobic bacteria. The highest bacteria isolation rate of Gram negative anaerobic bacteria could be detected in Porphyromonas and Prevotella. No metronidazole, cefoxitin, amoxicillin/carat acid resistant strain was found in the two Gram negative anaerobic bacteria. In the study, 48 patients with oral and maxillofacial infection were treated according to the results of drug resistance testing, and the clinical cure rate was 81.3%. Conclusion: Mixed aerobic and anaerobic bacteria cultures are very common in most oral and maxillofacial infection patients. Anaerobic bacteria culture and drug resistance testing play an important role in clinical treatment.

Objective：To understand the clinical features of osteonecrosis of the jaw after bisphosphonates use for therapy of breast cancer patients with bone metastasis. Methods: The cases diagnosed as bisphosphonates-related osteonecrosis of the jaws (BRONJ) were retrospectively analyzed from January 2011 to August 2015 in the Peking University School and Hospital of Stomatology, and those breast cancer patients with bone metastasis were selected. The clinical symptoms, imaging characteristics and treatment results were summarized. Results: A total of 14 cases of breast cancer patients with bone metastasis were selected, with an average age of 60.21 years. The average time of suffering from breast cancer was 9.77 years, and the average time of bone metastasis and bisphosphonates drugs use was 5.67 and 3.29 years individually. There was no patient with systemic application history of hormone therapy, and no history of diabetes. There were 9 patients with tooth extractions history, and the mean time of bone necrosis symptoms was 8.58 months. There were 10 cases with bone necrosis occurring on mandible, 3 cases on maxilla, and one case with both upper and lower jaws involved. Among the 10 patients with surgical treatment, there were 3 cases cured, and 6 cases improved. However, the clinical symptoms of 2 cases with conservative treatment were significantly aggravated. Conclusion: The medication time between the bisphosphonates use beginning and the occurrence of BRONJ is relatively long. The history of diabetes and long-time hormone use did not exist in this group. Tooth extraction itself does not determine the severity of BRONJ. Mandible is the most common site involved by BRONJ. Surgical treatment can alleviate the clinical symptoms of BRONJ with breast cancer to some extent.

Objective： To investigate the possibility for human papillomavirus (HPV) infection to be a predictable signal for the carcinogenesis of oral mucosa by comparing the prevalences of HPV in each stage of oral mucosal carcinogenesis and to compare the sensitivity differences of the two methods in detecting HPV infection in oral cavity.  Methods:The hybrid capture (HC-Ⅱ) was used to detect infection of HPV in 255 samples taken from 12 cases of healthy oral mucosa, 211 cases of patients with pathological diagnosis and 32 cases of patients with clinical diagnosis. The diagnosed cases included 8 cases of benign lesions of the oral mucosa, precancerous lesions ［74 cases of oral leukoplakia (OLK) with hyperplasia and 42 cases of OLK with oral epithelial dysplasia (OED)］, 91 cases of precancerous condition ［oral lichen planus (OLP)］ and 28 cases of oral squamous cell carcinoma (OSCC). And in situ hybri-dization (ISH) was used to detect infection of HPV in 33 cases of OSCC and 76 cases of OLK, including 30 cases of hyperplasia, 15 cases of mild OED, 15 cases of moderate OED and 16 cases of severe OED.Results: The prevalence of HPV in OLP samples was higher (12.12%, 8/66) than that of OLK (2.59%, 3/116) (χ2=4.666, P=0.031) and OSCC(7.14%, 2/28, χ2=0.513, P=0.474). The prevalence of HPV in OSCC (7.14%, 2/28) was higher than that of OLK (2.59%, 3/116), and no significant difference was found. There was only one case of smoke spot and statistical analysis was not carried out. ISH was used to detect type 16/18 and type 31/33 HPV DNA in 109 cases of oral mucosal lesions in paraffin sections and only one case of OSCC was HPV positive. Thirty-seven cases were detected by HC-Ⅱ and ISH methods at the same time. The same negative results by the two methods were found in 94.6% samples (35/37). In the other two samples, one was OSCC with early infiltration and the other was OLK with hyperplasia, The HC-Ⅱ results were positive while the ISH results were negative. The patients with OLP and HPV testing results were followed up and the average follow-up period was (36.2±10.5) months. It was found that three of them had a malignant transformation, and the malignant transformation rate of HPV positive patients was 12.50% (1/8), which was higher than that of HPV negative patients (3.45％, 2/58), and the difference was not statistically significant, P= 0.249.Conclusion: HC-Ⅱ assay was more sensitive in detecting HPV infection of oral mucosal lesions than ISH. The results of this study showed that there was insufficient evidence for taking HPV infection as a predictor of OLK carcinogenesis. Patients suffering from OLP were in a precancerous condition. The prevalence of HPV in OLP patients of this study was higher than that in OLK and OSCC patients, suggesting that for some reason, OLP patients were susceptible to HPV. HPV testing can be considered as routine in patients with OLP, and HC-Ⅱ assay was recommended. And patients with OLP and HPV positive should be followed up regularly.

Objective： To evaluate the vital signs changes, influence factors in different grades of hypertension patients during the treatment of acute pulpitis, in order to obtain the risk prevention measures. Methods: In this study, 90 different grades of hypertension patients with acute pulpitis were recruited from February 2014 to February 2015 in the Department of Oral Emergency, Peking University School and Hospital of Stomatology. The information about the patients’ general health, oral treatment, life signs of change information was collected. Patients were divided into high risk group, middle risk group, and low risk group (30 patients for each group). Results: (1) Compared with the preoperative, systolic blood pressure (90%), diastolic blood pressure (80%), heart rate increase (100%) were increased in the high risk group. The increase rates of the middle risk group and the low risk group were significantly lower than those of the high risk group (P<0.01). At the same time, the systolic blood pressure of 1/4 (26.7%) patients in high risk group increased more than 20 mmHg (1 mmHg=0.133 kPa), and the diastolic blood pressure of 2/5 patients in high risk group increased more than 10 mmHg, the difference was statistically significant compared with the other two groups (P<0.05). (2) Compared with the preoperative, the average increase of the maximum peak were increased ［systolic blood pressure (18.0±1.5) mmHg, diastolic blood pressure (8.0±1.7) mmHg］, the mean of heart rate changes ［(7.0±0.3) beats per minute］ was also increased in the high risk group, while these two indicators were decreased in the low risk group and the middle risk group. The electrocardiogram (ECG) was changed in 6 cases during the treatment in the high risk group. No significantly changed were observed in the low risk group and the middle risk group. (3) Compared the risk assessment in preoperative with that in postoperative, in the middle risk group, 23 cases were evaluated as medium risk in final evaluation, 6 as low risk, and 1 as high risk (risk assessment increased); in the high risk group, 20 cases were evaluated as high risk, 7 as very high risk, and 3 as medium risk (risk assessment decreased). Conclusion: Oral treatment is very safe for patients with hypertension, but the risk factor, target organ damage, and complications will also increase the risk of cardiovascular events in elderly patients during the acute pulpitis treatment. Dentist should take some measures to avoid the risks.

Objective：To evaluate the effect of the occlusal surface height of a mandibular posterior implant-supported single crown on stress in bone tissues. Methods: Three-dimensional finite element models of the implant-supported single crown replacing the missing right lower second premolar, mesial and distal natural teeth, periodontal membrane, alveolar bone, loaded rigid body and analog food of almond were established. Using the Federation Dentaire International （FDI） system, the first premolar, the second premolar and the first molar were represented with 44, 45, 46. Three occlusal surface heights of the crown were studied: (1) normal height; (2) 15 μm reduction in height; (3) 30 μm reduction in height. The models were loaded by independent loading with maximal occlusal force（44 by 280 N， 45 by 360 N， and 46 by 480 N）and average occlusal force（44 by 140 N， 45 by 180 N， and 46 by 240 N）on the single crown; combined loading (maximal occlusal force transformed into uniform load of 3.7 MPa on top of rigid body, in contact with points on the occlusal surface), and analog almondlike food loading (average occlusal force transformed into uniform load of 1.67 MPa in simulated food chewing, in contact with points on the occlusal surface). Results: For maximal biting force under independent loading, Von Mises stress peak values in bone tissues around 44, 45, and 46 were 82.57 MPa, 45.26 MPa and 27.79 MPa; For average biting force, peak values were 41.28 MPa, 22.63 MPa and 13.89 MPa. Under combined loading, compared with the normal occlusal surface height group, Von Mises stress peak values decreased 4.6 MPa, by 0.84%; increased 7.52 MPa, by 20.04%, and decreased 1.8 MPa, by 5.84%, for 45, 46, and 44 in the 30 μm infra-occlusion group, respectively. Under food loading, Von Mises stress peak values decreased 0.34 MPa, by 1.62%; increased 1.11 MPa, by 2.66%; and increased 0.06 MPa, by 0.54%, and for 45, 46, and 44 in the 30 μm infra-occlusion group, respectively. Conclusion:Within the limitation of this study, within 30 μm reduction of the occlusal surface height of implantsupported single crown, no significant difference of the peak values was observed.

Objective： To evaluate the efficacy and security of two type of rotary nickel titanium system (Twisted File and ProTaper Universal) for root canal preparation based on micro-computed tomography(micro-CT).Methods: Twenty extracted molars (including 62 canals) were divided into two experimental groups and were respectively instrumented using Twisted File rotary nickel titanium system (TF) and ProTaper Universal rotary nickel titanium system (PU) to #25/0.08 following recommended protocol. Time for root canal instrumentation (accumulation of time for every single file) was recorded. The 0-3 mm root surface from apex was observed under an optical stereomicroscope at 25×magnification. The presence of crack line was noted. The root canals were scanned with micro-CT before and after root canal preparation. Threedimensional shape images of canals were reconstructed, calculated and evaluated. The amount of canal central transportation of the two groups was calculated and compared.Results:The shorter preparation time ［(0.53±0.14) min］ was observed in TF group, while the preparation time of PU group was (2.06±0.39) min (P<0.05). In mid-root level, TF group shaping resulted in less canal center transportation than PU group ［(0.070±0.056) mm vs. (0.097±0.084) mm, P<0.05］. No instrument separation was observed in both the groups. Cracks were not found in both the groups either based in micro-CT images or observation under an optical stereomicroscope at 25× magnification.  Conclusion:Compared with ProTaper Universal, Twisted File took less time in root canal preparation and exhibited better shaping ability, and less canal transportation.

Objective： To study crown-root morphology of maxillary anterior teeth in Class Ⅱ, division 2 malocclusion using cone-beam CT (CBCT) combined with computer aided measurement technology to provide guidance for clinical treatment. Methods: The samples which consisted of 36 cases radiographed with CBCT techniques were selected and divided into two groups (18 each ) based on the type of malocclusion presented: Class Ⅱ, division 2 group (groupⅡ2) and Class Ⅰ group (groupⅠ). The measurements of crown-root morphology including crown-root angle and surface-shaft angle were got by Multiple Planer Reconstruction of CBCT data uploaded into InvivoDental software 5.0. The data were processed with SPSS 20.0 software package and t test was employed for comparison of angular measurements. Results: In group Ⅰ, crown-root angles of maxillary central incisor, maxillary lateral incisor and maxillary canines were 179.08°±3.31°, 176.55°±2.77°and 184.20°±2.51° respectively, surface-shaft angles were 21.00°±2.63°, 19.63°±2.35° and 19.36°±2.30° respectively. While in group Ⅱ 2, crown-root angles of maxillary central incisor, maxillary lateral incisor and maxillary canines were 176.80°±2.62°, 174.13°±3.28° and 181.79°±2.88° respectively, surface-shaft angles were 23.20°±2.95°, 22.29°±2.19° and 20.61°±2.34° respectively. Compared with group Ⅰ, significant statistical differences were observed with the exception of surface-shaft angle of maxillary incisor. There was significant difference in crown-root angle between group Ⅱ 2 and 180°.Conclusion: The maxillary anterior teeth in Class Ⅱ, division 2 malocclusion exhibited significant crown-root morphology which would influence the torque after orthodontic treatment. Special attention should be paid to the position of maxillary anterior teeth roots during orthodontic treatment for Class Ⅱ, division 2 malocclusion. The ideal position of tooth movement should be decided by the root rather than the location of the crowns.

Objective：To investigate the compensation of the anterior alveolar bone thickness in skeletal class Ⅲ patients treated with orthodontic-surgical treatment. Methods: The samples consisted of 54 ske-letal class Ⅲ patients treated with orthodontic-surgical treatment. Lateral cephalograms were taken before treatment. Descriptive statistics were calculated for corresponding variables, and the differences between the samples and the norms from Peking University normal occlusion sample library were assessed by independentsample t test. Correlation analyses were performed to find associations between skeletal characteristics and anterior alveolar bone thickness. According to skeletal anteroposterior discrepancy/vertical type (ANB, criteria=-4°; SN-MP, criteria=37.7°), the samples were allocated into group A (severe anteroposterior discrepancy/hypodivergent vertical type, n=11), group B (moderate anteroposterior discrepancy/hypodivergent vertical type, n=16), group C (severe anteroposterior discrepancy/hyperdivergent vertical type, n=14), and group D (moderate anteroposterior discrepancy/hyperdivergent vertical type, n=13),and one-way ANOVA with SNK multiple comparison test were performed. Results: The anterior alveolar bone thickness of the skeletal class Ⅲ patients were thinner compared with norm values (P<0.05). Correlational analyses showed that，both the upper and lower anterior alveolar bone thickness was correlated with the skeletal vertical discrepancy (P<0.05), but the anteroposterior type was only correlated with the lower anterior alveolar bone thickness (P<0.05). For the 4 groups according to skeletal anteroposterior discrepancy/vertical type, hypodivergent vertical type had thinner lower ligual and total bone thickness (LP, LW, P<0.05). while for upper alveolar bone thickness (UW), group C and group B exhibited the thinnest and thickest (the values of UW were 7.86 mm and 9.05 mm). Conclusion:The upper and lower anterior alveolar bone thickness of skeletal class Ⅲ patients are thinner compared with normal occlusion. Different skeletal anteroposterior discrepancy/vertical type results in differences in the anterior alveolar bone thickness, so decompensation should be treated differently and carefully.

Objective：To investigate the immediate bond strength and surface structure of resin and the tooth enamel which treated by cold plasma.Methods:In the study, 40 bovine incisors were divided into two equal parts. In this sense, all enamel adhesive samples were prepared and then randomly divided into 4 groups(n =20). group 1: acid + single bond 2+resin composite（control group）; group 2:beyond bleaching+ acid+single bond 2+resin composite; group 3: treated by cold plasma for 5 minutes+ acid+single bond 2+resin composite; group 4: treated by cold plasma for 5 minutes+single bond 2+resin composite. Single bond 2 bonding system and Filtek Z250 resin were used in this experiment. The shear bond strength was tested by universal testing machine. The surface of the enamel in different processes was observed by scanning electron microscope (SEM). Statistical analyses by the single factor analysis of variance and multiple pairwise comparisons were performed with SPSS 17.0 . Results: The shear bond strength of group 4 (8.60 MPa) was significantly lower than that of the other three groups (P<0.05). The shear bond strength of group 2 (17.89 MPa) was higher than that of group 4, but lower than group 1 and group 3 (P<0.05).There was no significant difference between group 1 (34.82 MPa) and group 3 (34.69 MPa). Scanning electron microscope indicated that the enamel treated by cold plasma had slight molten form, which was different from etched enamel surface.The fractured surface of group 3 was mix fracture, which was similar to the control group (group 1).Conclusion: Compared with the conventional clinic bleaching, immediate bond strength of resinenamel that treated by cold plasma has not been affected.

Objective： To evaluate the bone alteration subject to remodeling and analyze the esthetic result following immediate implant placement of incisors. Methods: In this study, 20 patients (13 women, 7 men) were involved, who needed implants for incisors of maxilla. The patients received 23 immediate implants totally. On the day of surgery and 6 months after the implants were placed, Cone beam CT (CBCT) was taken. The thickness of the alveolar ridge and the vertical change of marginal bone levels onthe mesial and distal aspects of theimplants were measured using the computer software  (PlanmecaRomexis Viewer 3.6.0.R). The evaluation of esthetic result by labial convexity score (LCS) and papilla index score (PIS) were analyzed pre-operation and one year after the final crown was delivered. The statistics with paired-t test for the measurement data and Willcoxon test for rating data were done by SPSS 20.0. Results: The survival rate in the two-year follow-up was 100%. The measuring point 1 (MP1), MP2, MP3 and MP4 (0, 2, 4, 6 mm apical to the implant platform, respectively) got significant alterations after 6 months of the follow-up. These differences were statistically significant (P<0.05).The major alteration happened at MP1 and MP4, which got (-0.89±2.06) mm and (-0.75±1.28) mm reduction of the alveolar, respectively. The marginal alveolar ridge resorption was （-0.42±1.24） mm and（-0.91±1.96） mm for Ankylos System and Replace System, respectively, and the difference was not statistical significant . The esthetic results were quite acceptable. Before treatment, 18 incisors rated 3 for LCS，and 2 incisors rated 4 for LCS；after final restoration, only 5 incisors rated 3 for LCS, and 14 incisors rated 2 for LCS. Before treatment, 15 incisors rated 3 for PIS; after final restoration, 13 incisors rated 3 for PIS. There was no statistically significant difference for the PIS pre-operation and 1 year after final restoration, while there was statistically significant negative change for LCS. Conclusion: Even following the proper surgical technique, the alveolar ridge wall still can’t be maintained after immediate implant placed in fresh socket of incisors. The inter-dental papilla could be well maintained, while due to the remodeling of labial bone, labial convexity will inevitably collapse. Therefore immediate implant still has esthetic risk.

Objective：To investigate the changes of the vertical height and width of the alveolar bone six months after the alveolar ridge preservation in periodontal compromised molar sites of severe alveolar bone defects with clinical direct measurement, parallel periapical radiographs, and cone-beam computed tomography (CBCT), and to analyze the effect of the three different methods of measurement. Methods: In this study, 20 subjects requiring tooth extraction on account of periodontal disease with a total of 23 extracted molars were enrolled. Extractions were performed atraumatically and patients were received alveolar ridge preservation procedure with Bio-Oss-and Bio-Gide-Clinical direct measurements were taken after tooth extraction and during the implant surgery 6 months later, CBCT scans and parallel periapical radiographs were taken immediately after ridge preservation and 6 months later. The changes of alveolar ridge width and vertical height after six months were measured and analyzed through the above-mentioned three methods and the similarities and differences of the measured effect were compared. Results: There were no significant difference of alveolar vertical height in the center of the extraction sites, the center of distal aspect, and distobuccal aspect between the clinical direct measurements and the CBCT measurements (P>0.05), alveolar vertical height in other points and alveolar width measurements were statically significant (P<0.05). After 6 months, 10 sites of 10 subjects were received a flap and reentered to perform dental implants surgery. The vertical height in the center of alveolar increased significantly and the changes of alveolar vertical height of clinical direct and CBCT measurement were （6.15±1.73） mm and （6.59±2.53） mm, respectively. The measurements of the width of the alveolar bone were （8.45±1.18） mm and （8.52±1.27） mm, respectively. The measurements of the two methods were not statistically significant (P>0.05). The change of the alveolar height in the center of the extraction socket after six months measured by parallel periapical was （5.84±4.28） mm, which was closed to the clinical direct measurement and the CBCT measurement. Conclusion: Clinical direct measurement and CBCT measurement were largely consistent in the evaluation of the alveolar bone height and width after the alveolar ridge preservation using deproteinized boving bone mineral (DBBM, Bio-Oss^R) and bioabsorbable collagen membrane (Bio-Gide^R) in periodontal compromised molar sites of severe bone defects.

Objective： To assess the clinical effects of carbon fiber reinforcement on the “All-on-Four” provisional prostheses. Methods:Provisional prostheses were divided into control group and carbon fiber reinforcing group according to whether carbon fiber reinforcement was used in the provisional prostheses base resin. In our study, a total of 60 patients (32 males and 28 females) with 71 provisional prostheses（28 maxilla and 43 mandible）were enrolled between April 2008 and December 2012 for control group; a total of 23 patients (13 males and 10 females) with 28 provisional prostheses (9 maxillas and 19 mandibles) were enrolled between January 2013 and March 2014 for carbon fiber reinforcing group. The information of provisional prostheses in the patients was recorded according to preoperative examination. We used the date of definitive prosthesis restoration as the cut-off point, observing whether fracture occurred on the provisional prostheses in the two groups. Additionally we observed whether fiber exposure occurred on the tissue surface of the provisional prostheses and caused mucosal irritation. The interface between the denture base resin and the fibers was examined using scanning electron microscopy (SEM). Results:The age ［(57.3±10.1) years vs.(55.1±11.4) years］, gender (32 males and 28 females vs. 13 males and 10 females), maxilla and mandible distributions (28 maxillas and 43 mandibles vs. 9 maxillas and 19 mandibles), the number of extraction jaws (46 vs. 23), the average using time ［(7.8±1.3) months vs. (7.5±1.1) months］, and the opposing dentition distributions of provisional prostheses of the patients showed no significant differences between the control and reinforcing groups. There were 21(29.6%) fractures that occurred on the 71 provisional prostheses in the control group; there was no fracture that occurred on the 28 provisional prosthesesin the carbon fiber reinforcing group. The fracture rate of the carbon fiber reinforcing group was significantly lower than that of the control group (P=0.001). No carbon fiber exposure and mucosal irritation were observed from clinical examination.SEM revealed relatively continuous contact between the fiber and acrylic resin, and the resin particles adhered on the surface of the carbon fibers. Conclusion: The addition of carbon fibers between abutments placed on “All-on-Four” provisional fixed denture base resin may be clinically effective in preventing “All-on-Four” denture fracture and can provide several advantages for clinical use.

Objective： To explore a method of constructing universal 3-dimensional (3D) colorized digital dental model which can be displayed and edited in common 3D software (such as Geomagic series), in order to improve the visual effect of digital dental model in 3D software. Methods: The morphological data of teeth and gingivae were obtained by intra-oral scanning system (3Shape TRIOS), constructing 3D digital dental models. The 3D digital dental models were exported as STL files. Meanwhile, referring to the accredited photography guide of American Academy of Cosmetic Dentistry (AACD), five selected digital photographs of patients’ teeth and gingivae were taken by digital single lens reflex camera (DSLR) with the same exposure parameters (except occlusal views) to capture the color data. In Geomagic Studio 2013, after STL file of 3D digital dental model being imported, digital photographs were projected on 3D digital dental model with corresponding position and angle. The junctions of different photos were carefully trimmed to get continuous and natural color transitions. Then the 3D colorized digital dental model was constructed, which was exported as OBJ file or WRP file which was a special file for software of Geomagic series. For the purpose of evaluating the visual effect of the 3D colorized digital model, a rating scale on color simulation effect in views of patients’ evaluation was used. Sixteen patients were recruited and their scores on colored and non-colored digital dental models were recorded. The data were analyzed using McNemar-Bowker test in SPSS 20-Results:Universal 3D colorized digital dental model with better color simulation was constructed based on intra-oral scanning and digital photography. For clinical application, the 3D colorized digital dental models, combined with 3D face images, were introduced into 3D smile design of aesthetic rehabilitation, which could improve the patients’ cognition for the esthetic digital design and virtual prosthetic effect. Conclusion: Universal 3D colorized digital dental model with better color simulation can be constructed assisted by 3D dental scanning system and digital photography. In clinical practice, the communication between dentist and patients could be improved assisted by the better visual perception since the colorized 3D digital dental models with better color simulation effect.

Objective： To explore the effect of fortified human milk feeding on growth and complications of infants with extremely and very low birth weight (ELBW/VLBW) during hospital stay by a prospective, random and controlled study. Methods: In the study, 122 ELBW/VLBW infants were enrolled and divided into two groups. The infants fed with human breast milk, combined with human milk fortification (HMF) during hospital stay were named HMF group (n=62), and those fed exclusively with premature formula were named premature formula feeding group (PF group, n=60). The data of the infants’ growth (the velocity of increase on the weight, length, head circumference and upper arm circumference), the time of rebounding to birth weight, the time of needing intravenous nutrition, the time of hospitalizing, the proportion of extrauterine growth retardation (EUGR) during hospital stay, the level of hemoglobin, bone metabolism and incidence of complications were compared between the two groups. Results: Among the 122 infants included, (1) the length increment in HMF group was higher than PF group ［(0.89±0.23) cm/week vs. (0.79±0.34) cm/week, P=0.04］; there were no significant differences in the weight gain, head circumference increment and upper arm circumference increment (P>0.05); (2) the age of rebounding to birth weight ［(10.13±4.03) d vs. (8.03±3.28) d, P=0.002］ and the duration of intravenous nutrition ［(16.77±6.63) d vs. (14.23±4.15) d, P=0.01］ in HMF group were longer than that in PF group, there were no significant differences between the two groups in the hospital stay and age achieved feeding; (3) there were no significant differences between the two groups in the incidence rate of EUGR during hospital stay (P>0.05); (4) the level of calcium at birth in HMF group was lower than that in PF group ［(2.19±0.22) mmol/L vs.( 2.32±0.27) mmol/L, P=0.005］, and the level of alkaline phosphatase (AKP) in HMF group at discharge was higher than in PF group ［(363.98±122.49) mmol/L vs. (299.73±117.39) mmol/L, P=0.004］; (5) the incidence of the feeding intolerance (6.5% vs. 18.3%, P=0.04) and sepsis (4.8% vs. 16.7%, P=0.03) in HMF group were less than in PF group, there were no significant differences between the two groups on the morbidity of necrotizing enterocditis, retinopathy of prematurity (ROP), and bronchopulmonary dysplasia (BPD) (P>0.05). Conclusion: HMF for premature infants may ensure the same growth pattern as those fed by premature formula, promote the calcium absorption, decrease the incidence of sepsis and feeding intolerance, and does not increase the incidence of necrotizing enterocolitis.

Objective： To evaluate the effectiveness and safety of endovascular treatment in solving symptomatic cerebral artery tandem lesions. Methods: From June 2012 to February 2014, 12 cases (24 lesions) with symptomatic cerebral artery tandem lesions were accepted for the endovascular treatment. The distributions of the tandem lesions were as follows: the common carotid artery and internal carotid artery (1 case), the internal carotid artery and the proximal of the carotid cavernous sinus segment (3 cases), the internal carotid artery and the distal of the carotid cavernous sinus segment (4 cases), the intracranial segment of internal carotid artery and middle cerebral artery M1 segment (2 cases), the first segment of vertebral artery and intracranial segment of vertebral artery (2 cases). All of these cases were treated from distal lesions to proximal lesions except for tandem lesions in the internal carotid artery and the distal of the carotid cavernous sinus segment in order to obtain better support. Tandem lesions were treated in the same operation with local anesthesia or general anesthesia. The procedures of the 12 cases retrospectively were analyzed and the peri-operation complications and responsibility region recurrent ischemic stroke incidents observed. Results:All tandem lesions were solved successfully all at once. There were no peri-operation complications or recurrent ischemic stroke incidents. There were no recurrent ischemic stroke incidents or stent restenosis cases in the followup.Conclusion:It is safe and effective for selective endovascular treatment in solving symptomatic cerebral artery tandem lesions at the same time, but we should take careful preoperative evaluation and improve the operation plan.

Objective：To design Keap1-tat peptide and explore its neuroprotective role on hipocampal CA1 neuron, as well as the effect on spacial learning and memory function following global cerebral ischemia. Methods: Adult male Sprague Dawley (SD) rats were subjected to global cerebral ischemia (GCI) by four-vessel occlusion for 15 min and randomly divided into five groups: sham, sham+Keap1-tat, ischemia/reperfusion (I/R), Keap1-tat peptide- and vehicleadministrated groups. For Keap1-tat or vehicle groups, the rats were treated with Keap1-tat (30, 50, 100 μg in 5 μL 0.9% saline) or the same vo-lume vehicle by intracerebroventricular injection (icv) 30 min prior to ischemia. Cresyl violet staining was used to observe the surviving neurons and 4-hydroxy-2-noneal (4-HNE) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) immunostaining were used to detect the change of markers response to oxidative stress in hippocampal CA1 region. The spatial learning and memory function of the rats was evaluated using Morris water maze. Results: Compared with sham group, the number of surviving neurons in ischemiareperfusion and vehicle groups significantly decreased in the hippocampal CA1 region (P<0.05), while administration of Keap1-tat significantly decreased the damage following GCI (P<0.05), and the dose of 50 μg existed the most effective neuroprotective role. Furthermore, immunostaining intensity of 4-HNE and 8-OHdG, markers of oxidative stress damage attenuated by Keap1-tat peptide as compared with vehicle group in CA1 region. Of significant interest, the time of finding underwater platform in Keap1-tat group animals was significantly short, and after removing the platform, the probe time of Keap1-tat group animals in the original quadrant where the platform was significantly increased compared with that of vehicle and I/R group animals (P<0.05). Conclusion: Keap1-tat peptide can effectively attenuate neuronal damage in hippocampal CA1 region and improve learning and memory function, which might bedue to the attenuation of oxidative stress caused by GCI.

Objective： To study the clinical effects of percutaneous transluminal angioplasty (PTA) versus stent implantation (ST) after PTA for the treatment of femoral and popliteal artery lesion resulted from arteriosclerosis obliterans. Methods: One hundred and three patients (119 limbs) treated for femoral and popliteal artery lesion resulted from arteriosclerosis obliterans for ten years were reviewed, of whom 60 limbs were treated by PTA and the other 47 by PTA combined with stent implantation.Results:Among the 60 limbs of the PTA group, there were 22 limbs involved only in femoral and popliteal artery; 13 limbs combined with iliac artery lesion; 17 limbs combined with infrapopliteal artery lesion; 8 limbs combined with iliac and infrapopliteal artery lesion. Among the 47 limbs of the ST group, there were 18 limbs involved only in femoral and popliteal artery; 8 limbs combined with iliac artery lesion; 15 limbs combined with infrapopliteal artery lesion; 6 limbs combined with iliac and infrapopliteal artery lesion. There was no significant difference between the two groups on age, sex, concomitant disease, ankle brachial index(ABI) before treatment and Rutherford classification (P>0.05). The patients’Trans-atlantic inter-society consensus (TASC) C/D was lower in the PTA group than that in the ST group (58.3% vs.76.6%, P=0.047).The follow-up periods were 48.0 (5.0,108.0) and 40.0 (3.0,96.0) months respectively (P=0.064).Compared with the PTA group, the ST group had a better short-term total effective rate (93.6% vs.80.0%, P=0.044) and a higher cost ［(33 882.7±8 695.6) yuan vs. (17 754.8±3 654.2) yuan, P<0.001］. The short-term marked effective rate of the ST group was higher than that of the PTA group, but the difference was not significant (31.9% vs.21.7%, P=0.231). There was no significant difference between the two groups on short-term efficiency, and complication rates (58.3% vs. 58.3%, P=0.724; 1.7% vs.2.1%, P=1.000). There was no death during perioperative period and no short-term deterioration in both the groups. The long-term marked effective rate was lower and the deterioration rate was higher in the ST group than that in the PTA group, but the difference was not significant (8.5% vs. 15.0%, P=0.381; 14.9% vs. 5.0%, P=0.081).There was no significant difference between the two group on long term total effective rate,accumulative limb salvage rate and reoperation rate (66.0% vs. 66.7%, P=0.939; 94.7% vs. 94.1%, P=0.884; 31.9% vs. 31.7%, P=1.000), and the 1 to 10 years primary and secondary patency rates were similar (P=0.837, P=0.622).When compared based on TASC classification, TASC A/B patients in the ST group had a higher short-term marked effective rate, a higher short-term total effective rate and a higher long-term deterioration rate than those in the PTA group, but the difference was not significant (36.4% vs. 24.0%, P=0.353; 100.0% vs. 88.0%, P=0.322; 18.2% vs. 4.0%, P=0.216). TASC C/D patients had a similar result (30.6% vs. 20.0%, P=0.307; 91.7% vs. 74.3%, P=0.050; 13.9% vs. 5.7%, P=0.226). Both TASC A/B and TASC C/D patients in the ST group had a similar accumulative limb salvage rate with that in the PTA group (90.9% vs. 90.6%, P=0.920; 97.1% vs. 94.1%, P=0.796). Conclusion: Stent implantation can increase the cost and short term effective rate at the same time and is not superior to PTA on the long term effective rate and limb salvage rate for femoral and popliteal artery lesion resulted from arteriosclerosis obliterans.

Objective：To investigate the normal sonographic anatomic characteristics of the pectoralis major and the clinical value of ultrasound in diagnosing the extent and location of the pectoralis major tears. Methods:High frequency transducer was used in scanning the pectoralis major. The ultrasonographic images of 40 normal pectoralis major were obtained from 20 healthy volunteers with both sides. Longitudinal and transversal views were performed and stored. The distal tendon was identified in the transverse plane coursing superficially to the long head of the biceps brachii tendon inferior to the level of the subscapularis tendon. Eighteen cases of pectoralis major tears were analyzed retrospectively，with MRI, surgical and ultrasound follow-up results correlation respectively. Results:High-frequency ultrasound could clearly show the anatomic orientation of the normal pectoralis major. The fibers converge was like a fan into three laminae that twisted upon each other at 90° before coalescing into a single tendon of insertion. In the study, 18 patients of pectoralis major muscle tears ［average age：(37.2±15.6) years］ sustained injuries during weightlifting, basketball and impact. Three of the eighteen patients had MRI results； nine had surgical correlation； six were followed by ultrasound. Eleven were injured on the left side, and 7 on the right side. Seven were involved in the distal tendon (1 in sternal head, 2 in clavicular head, 4 in both sternal and clavicular head), five were involved the musculotendinous junction, 6 were involved muscle belly. Twelve cases were partial-thickness petoralis major tears（4 in the distal tendon， four in the muscle tendon junction，4 in the muscle belly），with the partial fiber intact, echogenicity decreased and the internal structure disordered; 6 cases (3 in the distal tendon, 1 in the muscle tendon junction, 2 in pectoralis major muscle belly) were completely disrupted, with fiber fracture and retraction, accompanied with or without hematoma formation. Conclusion:Highfrequency ultrasound can clearly show the anatomic structure of the pectoralis major. Ultrasonography can diagnose the pectoralis major tears with the extent and location of injuries，and can be used to help the clinical treatment.

Objective：Human adipose-derived stem cells (hASCs) are a highly attractive source in bone tissue engineering. To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs) in vitro, and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vivo. Methods: The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OCpro-Luc-Puro construct. Then, the OCpro-Luc-Puro construct together with three assistant vectors: pMDLg/pRRE, pRSVREV, and pVSVG, were transiently transfected into HEK293T cells followed by viral supernatants collection, filtration and concentration. Next, the hASCs stably expressing luciferase repor-ter gene driven by osteocalcin promotor were created with the lentivirus carrying OCpro-Luc-Puro cassette under puromycin selection. The OCpro-Luc-hASCs were then cultured in the absence or presence of osteogenic differentiation medium. On the 7th and 14th days, after osteogenic induction, cellular extracts were collected and analyzed by luciferase reporter assay. Meanwhile, alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR) analysis of osteogenic associated genes osteocalcin (OC)， runt-related transcription factor 2 (Runx2) and  alkaline phosphatase (ALP) were used to assess the osteogenic differentiation ability of OCpro-Luc-hASCs. Results:OCpro-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated. On the 7th and 14th days after osteogenic induction, the luciferase activity of the cellular extracts from OCpro-Luc-hASCs was dramatically increased. Consistently, the extracellular matrix mineralization, as shown by Alizarin red S (ARS) staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC, Runx2 and ALP, although to variable extent, was observed upon osteogenic differentiation. These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OCpro-Luc-hASCs. Conclusion: We established a luciferase reporter system that could be used to rapidly, quantitatively and specifically determine osteogenic differentiation ability of hASCs. Comparing with the traditional time-consuming methods, the system we generated here was highly effective. This system not only can be used to examine ostogenic differentiation of hASCs in a high throughput manner, but also provides a way to monitor ostogenic differentiation of cells in vivo.

Objective：To introduce a novel ridge preservation technique with micro-titanium plate avoiding the use of bone grafting materials, and evaluate the potential horizontal bone preservation effect of this new technique, applied on single maxillary central incisors after tooth extraction for future implant restoration. Methods: Nine patients (six women and three men), mean age （26.0±5.7） years(from 18 to 34 years) referred to the Department of Oral Implantology, Peking University School and Hospital of Stomotology, were selected and diagnosed with unsalvageable single middle incisor with fine general conditions, no signs of acute local inflammation, no ongoing or previous periodontitis, healthy neighboring teeth and intact buccal bone walls. Tooth extraction, delayed implant placement and implant-supported single crown restoration were selected as treatment plan. The teeth were extracted atraumatically with local anesthesia, followed by a reflection of a minor flap to exposed 2-3 mm of the buccal bone plate. After that, a micro-titanium plate was trimmed and bended to fit the convexity of the labial bone and fixed by two mini pins with intent to support the labial soft tissue. The flap was then repositioned over the micro titanium plate and secured with two single sutures. No bone grafting materials or releasing incisions were needed. The sockets were left to heal without any intention of primary wound closure. Cone-beam computed tomographic (CBCT) scans were obtained before and four months after tooth extraction. Horizontal ridge widths were measured with CBCT software, and the preservation effects were calculated and recorded by the percentage of horizontal ridge alteration. Results: The nine extraction sockets were healed uneventfully. The average socket width before extraction was (7.51±0.48) mm (6.92-7.82 mm). The average alveolar ridge labial-palatal width at the control point of the edentulous area was (6.81±0.44) mm (6.04-7.38 mm) 4 months after tooth extraction, the mean percentage of ridge width preserved was 90.87%±2.91% (87.28%-95.60%). Conclusion: This novel ridge preservation method by the usage of a micro-titanium plate did not interfere with the natural socket healing process, and at the same time, largely preserved the width of alveolar ridge without any bone grafting procedures. Long term results remain to be seen.

As anorexia patients always go to the psychiatric clinic, little is concerned about the occurrence of sinus bradycardia in these patients for cardiologists and psychiatrists. The aim of this paper is to discuss the relationship between anorexia and sinus bradycardia, and the feature analysis, differential diagnosis and therapeutic principles of this type of sinus bradycardia. We report a case of sinus bradycardia in an anorexia patient with the clinical manifestations, laboratory exams, auxiliary exams, therapeutic methods, and her prognosis, who was admitted to Peking University Third Hospital recently. The patient was a 19-year-old female, who had the manifestation of anorexia. She lost obvious weight in a short time (about 15 kg in 6 months), and her body mass index was 14.8 kg/ m2. The patient felt apparent palpitation, chest depression and short breath, without dizziness, amaurosis or unconsciousness. Vitals on presentation were notable for hypotension, and bradycardia. The initial exam was significant for emaciation, but without lethargy or lower extremity edema. The electrocardiogram showed sinus bradycardia with her heart rate being 32 beats per minute. The laboratory workup revealed her normal blood routine, electrolytes and liver function. But in her thyroid function test, the free thyroid （FT）hormones 3 was 0.91 ng/L (2.3-4.2 ng/L)，and FT4 was 8.2 ng/L（8.9 -18.0 ng/L）,which were all lower; yet the thyroid stimulating hormone(TSH) was normal 1.48 IU/mL（0.55-4.78 IU/mL）. Ultrasound revealed her normal thyroid. Anorexia is an eating disorder characterized by extremely low body weight, fear of gaining weight or distorted perception of body image, and amenorrhea. Anorexia patients who lose weight apparently in short time enhance the excitability of the parasympathetic nerve, and inhibit the sympathetic nerve which lead to the appearance of sinus bradycardia, and functional abnormalities of multiple systems such as hypothyroidism. But this kind of sinus bradycardia and hypothyroidism have good prognosis. And asymptomatic sinus bradycardia with reversible causes, because of the great prognosis, they do not need special treatment. Multiple medical and psychiatric disciplines were consulted, and then, family care, nutritional support and psychiatric therapy were given, and she did not need thyroid hormone replacement therapy. The patient’s overall clinical status improved gradually during her hospital stay and her heart rate was recovered to 55 beats per minute.

Here we reported a patient with warfarin-induced autoimmune hepatitis (AIH), and explored new concerns for the pharmaceutical care of warfarin. A 57-yearold woman was admitted to hospital for repeated anorexia, abdominal pain and abnormal liver function. She received prosthetic heart valve replacement because of rheumatic heart disease, and had started warfarin medication since 2 years before. Her liver function was elevated with highest alanine aminotransferase 861 U/L, aspertate aminotransferase 604 U/L, and total bilirubin 106.7 μmol/L. Her anticoagulant therapy was switched to low molecular weight heparin and the liver function returned to normal. The liver function was elevated when she started to take warfarin again. The patient was then on liver protection therapy, and warfarin was stopped again for the liver biopsy for diagnosis reason. Through medication consultation and evaluation, pharmacists were invited to work together with the physicians and helped to differentiate the reason for abnormal liver function, and provided therapeutic suggestions. Also the pharmacists gained experiences in the treatment of AIH, and discovered a new and severe adverse drug reaction for warfarin. In treating this case, the pharmacists’ active involvement into the treatment and evaluation of the effect on the patient reflected the advantage and importance of the multidisciplinary cooperation for pharmacists and physicians when complex diseases are faced.