Objective:To observe the changes of plaque microflora during the experimental gingivitis and to analyze the relationship between the plaque microflora and the clinical parameters.Methods:11 young male subjects with healthy gingiva and without systemic disease were selected.Subgingival plaque samples(2 sites /person)were collected and then smeared by Gongo red at baseline(0 day),the 7th,14th,21st day(without oral hygiene)and 28th day(7 days after reestablishing oral hygiene)respectively during experimental gingivitis.At the same time the clinical parameters were recorded.The results of smear and the clinical parameters were analyzed.Results:The percentage of spirochete was the lowest at the baseline and increased during the experimental gingivitis,and then reached the highest level on the 21st day.The percentage of spirochete of the 21st day showed the significant different compared with that of baseline(P0.05)Conclusion:Spirochete is correlated to the development of the gingivitis.

Objective Through analyzing benign and malignant pleural effusion samples by proteomic techniques, finding protein biomarkers to provide help and new clues for effusion differential diagnosis.Methods Two-dimensional electrophoresis(2-DE) and matrix-assisted laser desorption ionization-time of flight-mass spectrometry(MALDI-TOF-MS) were used to search and identify protein biomarkers, enzyme linked immunosorbent assay(ELISA) was to validate the biomarkers.Results By comparing malignant group with benign group, 43 significantly different protein spots(Up or down regulated≥2 times) were found.Including 9 up regulated spots and 34 down regulated spots.And 7 spots were identified(Up or down regulated≥3 times) by MALDI-TOF-MS and validated 2 spots immunoglobulin λ(Igλ) and haptoglobin(Hp) by ELISA.The results showed that Igλ showed no statistical significance between two groups, while Hp showed the statistical significance(P<0.05).The diagnostic sensitivity and specificity of Hp in malignant pleural effusion were 75.00% and 52.38% at diagnostic cut-off point of 389.02 μg/L.Conclusion The application of proteomics technology has a great help with protein biomarkers searching in pleural effusion.HP has a certain value for the differential diagnosis of benign and malignant pleural effusionand and worthy of further study.

Objective To investigate the effect and potential mechanism of pigment epithelium derived factor(PEDF) acting upon SK-MES-1 cell and human umbilical vein endothelial cells(HUVECs).Methods CCK-8 was used to detect the effect of varying concentrations of PEDF upon HUVECs and SK-MES-1 cell, measuring the degree of cell proliferation and inhibition effect across varying times.The flow cytometry tests were carried out to invest gate the apoptosis of these two kinds of cells when exposed to varying concentration of PEDF.qRT-PCR were carried out to assess the vascular endothelial growth factor(VEGF) gene expression level in these two kinds of cells after treatment of PEDF.Results CCK-8 results revealed that PEDF had a concentration-dependent and time-dependent cell proliferation inhibition effect on SK-MES-1 cell and HUVECs(P<0.05);Flow cytometry showed that the apoptosis of the cells in the treatment group were higher than that of control group(P<0.05), and the apoptosis rate of high concentration group was higher than that of the low concentration group(P<0.05);qRT-PCR results showed that PEDF was able to inhibit expression of mRNA of VEGF in both HUVECs and SK-MES-1 cell compared with control samples(P<0.05).Conclusion The antitumor properties of PEDF is mainly related to the inhibition of tumor angiogenesis and direct effects on tumor cells, the effect of PEDF on HUVECs and SK-MES-1 cell maybe related to the effects of PEDF on downregulating expression of VEGF.

[Objective]To explore the clinic results of bi-posterolateral fusion combined with coralline hydroxyapatite interbody fusion,a three point stable spine fusion technique in the treatment of degenerative vertebra disc disorder.[Method]Bi-posterolateral fusion combined with posterior interbody fusion were performed to treat 41 cases of degenerative vertebra disc disorder from August 1998 to August 2006.The patients included 18 men and 23 women;their average age was 48 years.The slippage and grade of spondylolisthesis before and after spine fusion were investigated.The severity and slippage of radiographic measurements were also recorded,along with the average follow up of 19.1 month(range 12-48 months).The three point spine fusion applied autologous bone chips for bi-posterolateral fusion and one cubic block of coralline hydroxyapatite for anterior inter body fusion which achieved by PLIF technique.[Result]In the 41 patients who had underwent three point spine fusion surgery,only 1/41 patients experienced nerve root pain after surgery,and in most cases clinical symptoms were improved greatly.In 21/41 patients who had severe lower back pain completely disappeared after surgery,and in 9/41 cases these symptoms were minimal.The clinical evaluation according to the symptoms before the operation was excellent 2.4%(1/41),good 19.5%(8/41),fair 21.9%(9/41),bad 56.1%(23/41);after operation excellent 68.3%(28/41),good 29.2%(12/41),fair 2.4%(1/41),bad 0.0%(0/41).A radiographic measurements showed the anterior disc height was 8.05?2.35mm before operation and 11.44?3.38mm after operation(P

Objective: To measure GCF calprotectin and total protein for evaluating whether the calprotectin could be a sensitive marker for the initial inflammation of gingiva. Methods: Eleven young male subjects with healthy gingiva, who had no systemic diseases, were selected for this study. GCF samples (4 teeth /person) were collected with strips of filter paper at baseline (0 day), on the 7 th, 14 th, 21 st days (without oral hygiene), and 28 th day (7 days after reestablishing oral hygiene) during experimental gingivitis. The amount of calprotectin was measured by ELISA. The amount of total protein was assessed with protein dye binding assay. Results: The amount of calprotectin increased during the experimental gingivitis, and reached the highest level on the 21 st day. After oral hygiene was reestablished it reduced to the level of baseline. The amount of total protein had the same tendency as calprotectin. Conclusion: The amount of calprotectin and the total protein in GCF can reflect the initial inflammation of gingiva.

Diskoscope combined with screw-rod fixation was used in the treatment of lumbar spinal stenosis complicated by degree Ⅰ dislocation of 48 cases, with an average operation time 150 min, average amount of bleeding 160 mL, average admission for 14 days, and an average follow-up of 18 months. Four cases developed pain in lower limb caused by traction of nerve root, and the symptoms disappeared after symptomatic treatment. No loosening, nail breakage or rod breakage, no infection or inflammatory reaction was found after implantation. Clinical evaluation excellent rate was 95.8 % , indicating that diskoscope combined with screw-rod fixation after spinal fusion for the treatment of decompression has been effective, and the screw-rod system achieves an effective fixation, reduction of spondylolisthesis, and spinal stability.

BACKGROUND: As a bone graft, coralline hydroxyapatite (CHA) has uniform and interacted pore structure, which is suitable for vascular regeneration, bone regeneration and bone deposition. It displays good biocompatibility and no immunogenicity. OBJECTIVE: To explore the clinical results of CHA in the treatment of benign osteolytic bone defects. DESIGN, TIME AND SETTING: A retrospective analysis was performed in the Department of Orthopaedics, Affiliated Hospital of Hainan Medical College from May 1996 to May 2007. PARTICIPANTS: A total of 32 cases of benign osteolytic bone defects were enrolled from Department of Orthopaedics, Affiliated Hospital of Hainan Medical College. Pathological diagnosis showed that the cause of bone defect included bone cyst for 8 cases, fibrous hyperplasia of bone for 9 cases, aneurysmal bone cyst for 4 cases, osteoenchondroma for 1 case; fracture complication for 5 cases consisting of femoral complete fracture for 2 cases and humeral complete fracture for 1 case, and humeral incomplete fracture for 2 cases. CHA was prepared by the Biomaterials Laboratory of Hainan Medical College. METHODS: According to routine approach of the operation, 32 cases of benign osteolytic bone defects were implant with CHA granules, chips or blocks after deleting tumor tissue and thinning cortical bone. Then the periosteum was sutured. Three cases with complete fracture received internal fixation, while other cases were untreated. None was fixed with plaster for external fixation. MAIN OUTCOME MEASURES: The X-ray films were observed to evaluate the bone healing at different time. RESULTS: All cases were followed up for average 6-24 months. No general abnormal reactions were found. The incisions were healed in two weeks. The lesion range of bone defect was from 3 cm?2 cm?2 cm to 12 cm?4 cm?4 cm before operation; At 1-3 months after operation, bone graft began to fuse with bone tissue around defects and fused completely at 3-6 months, which indicated the bone defect was almost repaired; At 6-24 months, bone graft was moulded and rebuilt, gradually substituted by newly formed born. CONCLUSION: The CHA is an idea bone graft substitute material for its good results in the treatment of benign osteolytic bone defects, shorter operating time and fewer complications.

Objective: To investigate the changes of osteoprotegerin ( OPG) and receptor activator of nuclear factor kappa B ligand ( RANKL) level in perio-implant crevicular fluid ( PICF) and to monitor the development of the stability of Straumann ? tissue-level implants by resonance frequency analysis ( RFA) during the early phases of healing .Methods: A total of 35 implants ( length 10 mm ) were placed.PICF samples were collected with filter paper strips at baseline , 1, 2, 3, 4, 6, 8, and 12 weeks post-surgery, respectively.The OPG, RANKL levels were determined by ELISA method .At the same time points, the implant stability quotient (ISQ) values were determined with Osstell TM mentor.Results:During healing , PICF-OPG levels increased significantly 2 weeks after surgery when compared with the 4th-, 6th-, 8th-and 12th-week reevaluation (P<0.05).The OPG/RANKL ratio in PICF was significantly higher ( P<0 .05 ) than that in gingival crevicular fluid at 1 week post-surgery .ISQ slightly fluctuated within the first 4 weeks after installation .Following this, the ISQ values increased steadily for all the implants and up to 12 weeks.Significant differences were noted between the mean ISQ values at the 12th-week and other observation time points .Conclusion: The PICF-OPG levels may be effective in monito-ring the process of osseointegration .All the ISQ values indicated the stability of Straumann ? implants over a 12-week healing period .RFA is a reliable and effective assistant to monitor implant stability .

Objective: To evaluate methods of detecting various component in the trace sample of the gingival crevicular fluid (GCF). Methods: One sample of GCF was collected from four patients with mild or moderate periodontitis. At first, one half (group A)of every sample was measured for interleukin-6 (IL-6) by enzyme-linked immunosorbent assay (ELISA) and then detected for Interleukin-1? (IL-1?) in the same sample .Another half (group B) of the sample was used as control for the measurement of Interleukin-1? (IL-1?). Another sample of GCF was collected from nine patients with abutment and non-abutment because of the distal extension teeth lost resulted from the chronic periodontitis. At first, one half (group C) of the sample was detected for tumor necrosis factor -?(TNF-?) by ELISA and then measured for elastase in the same sample. Another half (group D) of the sample was used as control for the measurement of elastase. Results: There was no significant difference of both the measured value of IL-1? absorbency (group A and B are 0.5?0.4 and 0.5?0.4, respectively, P=0.136)and that of elastase absorbency (group C and D are 1.1?0.6 and 1.1?0.6,respectively, P=0.943) between the original elution and the sample after detection of IL-6 or TNF-? in the test 1 and test 2, respectively. Moreover, the values showed high correlation (r=0.993, P=0.000 in the group A and B; r=0.979, P=0.000 in the group C and D). Conclusion: It is practicable that one sample of GCF can be reused to examine for several components of GCF.

AIM: To explore whether tumor necrosis factor-α (TNF-α) induces necroptosis in murine long bone osteocyte-like cell line MLO-Y4 and the possible mechanism.METHODS: The MLO-Y4 cells were divided into control group, TNF-α group, TNF-α+necrostatin-1 (Nec-1) group, TNF-α+Z-VAD group and TNF-α+receptor-interacting protein 3 (RIP3)-siRNA group.The death rate of MLO-Y4 cells was assessed by flow cytometry with Annexin V-FITC/PI staining.The morphological features of the cells were observed under transmission electron microscope (TEM).The protein levels of RIP1, RIP3 and cleaved caspase-3 were determined by Western blot.Finally, the numbers of total cells and RIP1-RIP3-positive cells were observed under laser scanning confocal microscope.The production of reactive oxygen species (ROS) in the cells was measured by DCFH-DA staining.RESULTS: Compared with control group, the apoptotic or necroptotic rate of the cells induced by TNF-α was increased significantly (P<0.01).The increased apoptotic or necroptotic rate was dramatically reduced by treating with Nec-1, Z-VAD or RIP3-siRNA transfection (P<0.01).In TNF-α group and TNF-α+Z-VAD group, a lot of MLO-Y4 cells with typical necroptotic morphological features were observed under TEM.However, obvious necroptotic cells were not found in Nec-1 or RIP3-siRNA treatment group.The protein level of RIP1 in the cells treated with Nec-1 was sharply lower than that in TNF-α group (P<0.01).However, Z-VAD did not reduce the elevated levels of RIP1 and RIP3.RIP3-siRNA effectively down-regulated the protein level of RIP3 compared with TNF-α group (P<0.01).Nec-1 effectively down-regulated the protein levels of RIP1 colocalized with RIP3 compared with TNF-α group (P<0.01).However, Z-VAD did not reduce the levels of RIP1 colocalized with RIP3.Nec-1, Z-VAD and RIP3 siRNA significantly decreased the ROS levels (P<0.01).CONCLUSION: TNF-α induces the necroptosis of MLO-Y4 cells.RIP3 play vital roles in the cell necroptotic signal pathway.ROS may be the executor of necroptosis of MLO-Y4 cells.