OBJECTIVETo assess the efficacy of tendons of minimally invasive therapy (TMIT) combined drug therapy by comparing it with treatment by drug therapy alone on patients with knee osteoarthritis (KOA).

METHODSTotally 60 KOA patients were assigned to the treatment group and the control group according to random digit table, 30 in each group. Patients in the control group took Hydrochloric Acid Glucosamine Capsule and Celecoxib Capsule. Patients in the treatment group additionally received TMIT. The treatment course for all was 4 weeks. Scores for visual analogue scale (VAS) and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index were observed and recorded at week 1 and 4 after treatment by acupotomology mirror.

RESULTSCompared with before treatment, improvement was shown in VAS score, pain and stiffness degrees, activities and functions, and WOMAC scores at week 1 and 4 after treatment in all patients with statistical difference (P < 0.05). Besides, better effect was shown in the treatment group (P < 0.05).

CONCLUSIONSTMIT combined drug therapy could relieve KOA patients' pain, stiffness and joint activities, elevate the overall efficacy. TMIT was easily operated with less injury.

Celecoxib ; Drug Therapy, Combination ; methods ; Humans ; Knee Joint ; Osteoarthritis, Knee ; drug therapy ; Pain ; Pain Measurement ; Tendons ; Treatment Outcome

Adult ; Bone Neoplasms ; diagnostic imaging ; therapy ; Combined Modality Therapy ; Curettage ; methods ; Embolization, Therapeutic ; methods ; Female ; Follow-Up Studies ; Giant Cell Tumor of Bone ; diagnostic imaging ; therapy ; Humans ; Male ; Sacrum ; Time ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate effect of root canal curvature and location of the fragment on the removal of broken file from root canal.

METHODSSixty extracted mandibular premolars were divided equally into six groups according to location of fragment (2 mm or 8 mm below root canal orifice) and root canal curvature (20 degrees, 30 degrees or 40 degrees). Broken files were removed using ultrasonic tips combined with dental operating microscope. Number of successfully removed case and operating time were recorded. Pre- and post-operative digital radiographs were input into image analyzing software to calculate diameter variance of root canal at the level of tip of broken file.

RESULTSAll the 6 groups of broken files were successfully removed with out perforation. Among same fragment location groups, diameter variance was increased in greater curvature groups and the difference was significant (P < 0.05), while operation time was not significantly different (P > 0.05). Among same root canal curvature groups, the deeper the fragment, the longer operation time, difference being significant (P < 0.05), while difference of diameter variance was not significant (P > 0.05).

CONCLUSIONThe more curvature of root canal, the more dentine removal amount at the level of broken file tip. Location of fragment has no effect on dentine removal amount. Safe straight-line access and adequate thickness of dentine are essential to removal of broken file from root canal.

Bicuspid ; Dental Pulp Cavity ; Dentin ; Humans ; Root Canal Preparation ; Root Canal Therapy

OBJECTIVETo evaluate the activity of DNA polbeta promoter on p53 gene in salivary adenoid cystic carcinoma (SACC) cells.

METHODSThe luciferase activity was examined and used to evaluate the activity of DNA polbeta promoter on SACC-83 cells. Eukaryotic expression plasmids of p53 gene were constructed and stably transfected into SACC-83 cells. RT-PCR was used to assess the expression of p53 gene. The SACC-83 cells were subjected to the treatments of H2O2, ultraviolet radiation, Bleocin, and affected p53 mRNA and protein level in SACC-83 cells were characterized with RT-PCR and Western blotting.

RESULTSThe result of luciferase activity proved that the activity of DNA polbeta promoter in SACC-83 cells was much higher than that of CMV promoter. The results of RT-PCR suggested that p53 gene with different promoters were all expressed effectively, but the expression efficiency was different. It was greater in DNA polbeta group than in CMV group. After DNA damage, p53 gene expression increased and DNA polbeta promoter could enhance the expression of p53 gene more than CMV promoter. The results of Western blotting indicated that the expression of P53 protein between the two groups did not show any difference.

CONCLUSIONIn SACC cells, the activity of DNA polbeta promoter was increased and DNA polbeta promoter could enhance the expression of p53.

Carcinoma, Adenoid Cystic ; Cell Line, Tumor ; DNA ; Genes, p53 ; Humans ; Hydrogen Peroxide ; Promoter Regions, Genetic ; RNA, Messenger ; Salivary Gland Neoplasms ; Transfection ; Ultraviolet Rays

OBJECTIVETo develop a novel scaffolding method for the copolymers poly lactide-co-glycolide acid (PLGA) to construct a three-dimensional (3-D) scaffold and explore its biocompatibility through culturing Schwann cells (SCs) on it.

METHODSThe 3-D scaffolds were made by means of melt spinning, extension and weaving. The queueing discipline of the micro-channels were observed under a scanning electronic microscope (SEM).The sizes of the micropores and the factors of porosity were also measured. Sciatic nerves were harvested from 3-day-old Sprague Dawley (SD) rats for culture of SCs. SCs were separated, purified, and then implanted on PLGA scaffolds, gelatin sponge and poly-L-lysine (PLL)-coated tissue culture polystyrene (TCPS) were used as biomaterial and cell-supportive controls, respectively. The effect of PLGA on the adherence, proliferation and apoptosis of SCs were examined in vitro in comparison with gelatin sponge and TCPS.

RESULTSThe micro-channels arrayed in parallel manners, and the pore sizes of the channels were uniform. No significant difference was found in the activity of Schwann cells cultured on PLGA and those on TCPS (P larger than 0.05), and the DNA of PLGA scaffolds was not damaged.

CONCLUSIONThe 3-D scaffolds developed in this study have excellent structure and biocompatibility, which may be taken as a novel scaffold candidate for nerve-tissue engineering.

Animals ; Biocompatible Materials ; Cell Adhesion ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Lactic Acid ; Microscopy, Electron, Scanning ; Polyglycolic Acid ; Rats ; Rats, Sprague-Dawley ; Schwann Cells ; cytology ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo investigate the effects of the hepatitis B virus (HBV) P22e protein on the apoptosis of human hepatocellular carcinoma HepG2 cells.

METHODSHepG2 cells were transfected with recombinant plasmid pEGFP-HBVP22e and exposed to Act-D and tumor necrosis factor alpha (TNFalpha) treatment to induce cell apoptosis. Flow cytometry was performed to determine the proportion of cells containing sub-G1 DNA to represent the number of apoptotic cells. Laser scanning confocal microscopy was used to observe the nuclear alterations in the apoptotic cells.

RESULTSHepG2EGFP-C2HBVP22e cell strain showed a much delayed apoptosis as well as obviously lowered apoptotic rate in comparison with the HepG2 strain (P<0.01).

CONCLUSIONThe introduction and expression of extraneous gene HBVP22e significantly inhibits the apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Humans ; Transfection ; Viral Core Proteins ; metabolism

To study the effect of micro (mi)RNA on cellular proliferation induced by hepatitis B x protein, HBx, in human liver cells and to investigate the underlying molecular mechanism of this cancer-related effect. The human L02 hepatocyte cell line was stably transfected with HBx (L02/HBx) or an HBx mutant (L02/HBx-d382) that induces higher levels of cellular proliferation. The differential miRNA expression profiles were determined by microarray analysis and confirmed by real-time PCR. Two miRNAs, miR-338-3p and miR-551b, that were found to be significantly down-regulated in the L02/HBx-d382 cells were selected for further study and transfected individually into cells using the lipofectamine procedure. The cell survival rate was analyzed by MTT assay, and cell cycles were assessed by flow cytometry. Expressions of cyclinD1, cyclinG1, and E2F1 were assessed by real-time PCR and Western blotting. Compared with the microarray miRNA profile of L02/pcDNA3.0 cells, six miRNAs were up-regulated and five miRNAs were down-regulated in the L02/HBx-d382 cells, while four miRNAs were up-regulated and 12 were down-regulated in the L02/HBx cells. The microarray results were consistent with real-time PCR results. Transfection of miR-338-3p and miR-551b significantly inhibited the cell survival rates (P less than 0.001) and induced G0/G1 phase cycle arrest. According to MTT results: for L02/HBx-d382 cells, compared with lipofectamine or non-transfected (NC) controls, the t value of miR-338-3p was 10.402, 9.133 and the t value of miR-551b was 8.763, 7.403; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 9.105, 8.074 and the t value of miR-551b was 7.673, 7.52. According to flow cytometry results: for L02/HBx-d382 cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 12.173, 11.107 and the t value of miR-551b was 15.364, 13.377; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 15.416, 13.378, and the t value of miR-551b was 13.276, 13.109. The protein levels of cyclinD1, cyclinG1, and E2F1 were significantly reduced by both miR-338-3p and miR-551b ( P less than 0.001). For L02/HBx-d382 cells, compared with lipofectamine or NC controls: E2F1 had t = 11.132, 10.031 and 12.017, 10.973, respectively; cyclinD1 had t = 15.654, 15.013 and 15.447, 14.733, respectively; cyclinG1 had t = 8.017, 7.661 and 7.402, 7.417, respectively. For L02/HBx cells, compared with lipofectamine or NC controls: E2F1 had t = 14.244, 13.331 and 15.022, 14.468, respectively; cyclinD1 had t = 8.695, 8.137 and 7.877, 7.503, respectively; cyclinG1 had t = 7.73, 7.471 and 7.596, 7.41, respectively. In contrast, the mRNA levels for E2F1, cyclinD1, and cylcinG1 showed no significant differences between the miRNA transfected cells and controls. Wild-type HBx and the high proliferation-inducing mutant HBx can influence the miRNA expression profile of L02 cells. HBx down-regulates miR-338-3p and miR-551b in L02 cells, and the high proliferation-inducing mutant has a more robust effect. The mechanism of miR-338-3p- or miR-551b-mediated cell growth inhibition appears to be related to the direct modulation of cyclinD1, cyclinG1, and E2F1.
Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line ; Cell Proliferation ; Cyclins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, Viral ; Hepatitis B virus ; genetics ; metabolism ; Hepatocytes ; metabolism ; pathology ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Mutation ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; Transfection

3a and 7a are nonstructural proteins of SARSCoV, which are encoded separately by ORF 3a and ORF 7a in SARSCoV genome. The expression of 3a has been founded in cells infected by virus in vivo or in vitro. Firstly, the pGL3Control vector was reconstructed , the pGL3Enhancer vector deletious of SV40 promoter gene was obtained . Then the IFN? promoter gene was cloned into the pGL3Enhancer vector and pGLIP21, the Luciferase reporter plasmid with IFN? promoter was established. The availability of pGLIP21 was verified by NDV ,the inductor of IFN?, the Luciferase activity was assayed in cells transfected with pGLIP21 by Luminometer. In order to see the function of 3a and 7a protein of SARSCoV,CHO cells expressing 3a or 7a protein were transfected with pGLIP21, the intensity of luciferase activity was analyzed . By analysis, in vitro, 3a and 7a protein of SARSCoV had the similar ability in triggering the expression of Luceferase gene, i.e 3a and 7a protein of SARSCoV could effectively activate the promoter fragment of IFN? gene. This result will help studying the function of 3a and 7a protein and provide a method to study the nosogenesis mechanism of SARSCoV.

Objective To explore the significance of designing with monitoring-flap in massive com- pound bone grafts for repairing massive bone defects in extremities.Methods From January 2001 to De- cember 2004,large bone defects in 19 patients(11 men and 8 women,age:6 to 35 years,mean age:18.6 years)were repaired by vascularized free fibular transplant with a monitoring-flap combining with massive deep frozen bone allografts.Average length of the bone defects was 16.6 cm(range,12 to 25 cm).A 7 days' con- tinuously clinical examination including observing the color,turgor,temperature,capillary refill,and bleeding after a needle sticking of the monitoring-falps were used postoperatively,if any one of these were abnormal,the circulation of the compound bone grafts must be in danger and some measures such as re-operation should be taken immediately.Dynamic image analysis was used for evaluating the bone union.Results One monito- ring-flap was vascular artieulo,and the articulo was relieved after exploration and resection of vein thrombus; another one was marginal part necrosis;the remains were normal.All of monitoring-flaps healed normally after 23.2 months(range,6 to 54 months)follow-up.15 patients had the radiographic evidence of bone unions 3 months after surgery.11 patients had been removed intermal fixation,complete bone unios were found one year postoperatively.Conclusion Designing with monitoring-flap in massive compound bone grafts for repairing massive bone defects,and can clearly understand the circulatory statue of compound bone grafts and early pre- dict the final results of massive bone allografts.

OBJECTIVETo evaluate the inhibitory effect of p53 gene on salivary adenoid cystic carcinoma (SACC) cells.

METHODSAdenoviral vector pDeltaE1-p53 was constructed and transfected into SACC-83 cells. The enhanced p53 expression was measured by reverse transcription polymerase chain reaction (RT-PCR), and the effects of transfected p53 on SACC-83 cells were analyzed by TRAP-PCR-ELISA, luciferase reporter, flow cytometry (FCM), soft agar assay and tumorigenicity test.

RESULTSThe expression of p53 gene in SACC-83 cells was increased after introduction of pDeltaE1-p53. The telomerase activity and the transcriptional activity of hTERT promoter were inhibited. The cells cycles of transfected SACC-83 were arrested in G(1) phase and the rate of colony-formation was decreased, and similarly the tumorigenicity in nude mice was also decreased.

CONCLUSIONSThe introduction of wild-type p53 by adenoviral vector could suppress the telomerase activity and malignant phenotypes of salivary adenoid cystic carcinoma cells.

Animals ; Carcinoma, Adenoid Cystic ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Genetic Vectors ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Salivary Gland Neoplasms ; genetics ; metabolism ; pathology ; Telomerase ; genetics ; metabolism ; Transfection ; Tumor Suppressor Protein p53 ; genetics