Objective To detect the expression and effect of human tumor necrosis factor related apoptosis-inducing ligand (hTRAIL) in vivo,by using a novel double expressing adenoviral vector encoding hTRAIL and firefly luciferase ( luc ) gene ( ad -luc-hTRAIL),in which luc was used as reporter gene.Methods Lung cancer A549 cell xenografts in 16 nude mice models were established in subcutaneous inoculation way,the adenovirus vectors ( ad-luc-hTRAIL,ad-hTRAIL,ad-luc) and phosphate buffer saline (PBS) (n =4) as control were injected into tumor respectively.The size of the tumor was measured at different time points (4,7,10,14,21,28 d)after injection.The activity of luciferase in surface of the tumor was detected in vivo by using high-sensitivity cooled-charged coupled device(CCD) camera.The expression of hTRAIL was demonstrated by immunohistochemistry staining after sacrificing the animals at different time points,and immunohistochemical scores (IHS) were measured. The apoptosis rate of tumor cells was detected by using TUNEL and calculated. Analysis of variance,the paired t test and linear correlation analysis was used for the statistics. Results The growing speed of tumour xenografts was more slowly in ad-luc-hTRAIL and ad-hTRAIL groups than PBS group (t =2.71,2.72,P ＜ 0.05 ).The tumor volumes of ad-luc-hTRAIL,ad-hTRAIL,ad-luc and PBS groups 28 days after injection were (208.4 ± 42.3 ),( 181.5 ±23.9),( 403.1 ± 54.0 ) and ( 427.0 ± 59.3 ) mm3, respectively. There was no significant difference between ad-luc group and PBS group(t =2.07,P ＞ 0.05).The expression of luciferase in ad-luc-hTRAILgroup reached its peak at 7th day ( 1.37 ± 1.04),and then decreased quickly.The IHS and apoptosis rate in ad-luc-hTRAIL and ad-hTRAIL groups reached their peaks at 7th day,the peak values of IHS were 6.25 ±2.06 and 6.5 ± 2.89,the peak values of apoptosis rate were (60.75 ± 8.06 ) ％ and ( 61.50 ± 8.47 ) ％,respectively.The amount of luciferase expression ( absolute number of photons detected by CCD camera)was linear positive correlated with IHS and apoptosis rate ( rphotons/IHs =0.942,rph /rate =0.842,rIHs/rate =0.887,P ＜ 0.05 ).Conclusion The target gene hTRAIL can be transfected into lung cancer A549 cell xenografts nude mice models efficiently with a high level expression,and the therapeutic effect of hTRAIL can be monitored by detecting the expression of luc.

This study was aimed to establish the three-dimensional model of C_(3-7) segment of lower cervical spine after artificial disc implantation, to analyze the movement of lower cervical spine after artificial disc replacement. According to CT films of 1 patient at 6 months after artificial disc implantation, three-dimensional finite element model that included Bryan~(TM) artificial cervical disc prosthesis of the lower cervical spine was established using finite element method, then introduced into Ansys 9.0, the vertebral cortical bone, cancellous bone and intervertebral disc were meshed and analyzed by using under several states such as flexion/extension, lateral bending and rotation, thus understanding their motion characteristics. By comparison with previous research findings, test results nearly accorded with or exhibited identical trend with previous study. The results suggest that, cervical disc replacement can basically guarantee the stability of lower cervical spine movement.

The paper analyzes the difference of higher stomatological education in China and abroad and the weakness in the teaching system of China,and proposes some reform consideration based on our real situation.

Objective To analyze the medication rules of the treatment of stroke in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan; To provide references for the clinical treatment. Methods Prescriptions with confirmed efficacy of famous TCM doctors in the history in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan was searched. Excel2003 was used to establish database to analyze medication frequency. SPSS17.0 statistical software was used to conduct cluster analysis, and tree view was used to show results. Results Totally 112 prescriptions for the treatment of stroke of 36 famous TCM doctors were included in the study, including 204 kinds of Chinese materia medica and 1169 times of medication frequency. The high-frequency medicines (>10 times) were Paeoniae Radix Alba, Achyranthis Bidentatae Radix, Angelicae Sinensis Radix, Pinelliae Rhizoma Praeparatum, and Poria. The high-frequency medicine categories were tonifying deficiency medicine (22.58％), pacifying liver and wind medicine (12.31％), activating blood and dispelling stasis medicine (11.89％), clearing heat medicine (11.46％) and dissipating phlegm, cough and asthma medicine (8.72％). Cluster analysis showed that high-frequency medicine (>10 times) could be clustered as 6 categories. Conclusion Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan focuses on tonifying deficiency medicine, accompanied with pacifying liver and wind medicine, activating blood and dispelling stasis medicine, clearing heat medicine and dissipating phlegm, cough and asthma medicine, which can be used to guide clinic.

OBJECTIVETo determine the optimized condition under which BMP expression vector will be constructed to transfect bone marrow stromal cells (MSCs), plasmid vector coding enhanced green fluorescence protein (EGFP) gene pEGFP was transferred into MSCs. The transfer efficiency and transient expression were subsequently tested.

METHODSpEGFP plasmid was amplified and tested by an enzyme cutting technique in vitro. MSCs, which were initially obtained from the bone marrow of rabbits, were cultured in vitro and transferred with pEGFP by means of lipofectamine media methods. The ratio of plasmid and lipofectamine was varied according to the experiment design. Transfer efficiency and transient expression were evaluated by fluorescent microscopy.

RESULTSTransfer efficiency was correlated with the ratio of plasmid and lipofectamine. The expression of EGFP began in 24 hours after transferring, reached maximum in 48-72 hours and decreased in 1 week, however there remained a weak expression for more than 3 weeks.

CONCLUSIONThe efficiency of transferring pEGFP into MSCs could achieve to 30% with proper ratio of plasmid and lipofactamine. pEGFP was an ideal transient expression vector for MSCs gene transference.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Bone Morphogenetic Proteins ; genetics ; Cells, Cultured ; Genetic Vectors ; Green Fluorescent Proteins ; Lipids ; genetics ; Luminescent Proteins ; biosynthesis ; genetics ; Plasmids ; genetics ; Rabbits ; Stromal Cells ; cytology ; metabolism ; Transfection

OBJECTIVETo investigate the role of HPV16E6 and E7 during the transformation of oral epithelial cells.

METHODSAn human immortalized oral epithelial cell line (HIOEC) was established by transfecting HPV16E6, E7 open reading frames using recombinant retroviral system plxsn to human normal oral epithelial cells. Expression of HPV16E6, E7, Rb, P16 and Cycin D1 were analyzed by Western blot in HIOEC and human normal oral epithelial cells. Formation of complex of HPV16E7 and Rb were analyzed by Immunoprecipitation-western blot. Human normal oral epithelial cells and the oral epithelial cells transfected with plxsn were used as control groups.

RESULTSHIOEC expressed HPV16 E6 and E7; HIOEC expressed both hyperphosphorylated and underphosphorylated Rb while oral epithelial cells in two control groups only expressed hyperphosphorylated Rb. HPV16 E7 formed complex with underphosphorylated Rb; the level of P16 and Cyclin D1 had no remarkable change.

CONCLUSIONSHPV16E7 plays an important role in the immortalization of oral epithelial cells induced by HPV16.

Blotting, Western ; Cell Line ; Cell Transformation, Neoplastic ; Cyclin D1 ; analysis ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Humans ; Mouth Mucosa ; metabolism ; pathology ; virology ; Oncogene Proteins, Viral ; physiology ; Papillomavirus E7 Proteins ; Phosphorylation ; Repressor Proteins ; Retinoblastoma Protein ; analysis

OBJECTIVETo establish an immortalized oral epithelial cell line.

METHODSNormal human oral epithelial cells were transfected with HPV16E6/E7 open reading frames using recombinant retroviral system pLXSN. Expression of HPV16E6 and E7 protein were tested by Western blot in three kinds of cells. To define cellular biological characterization of HPV16E6/E7 transfected cells, a series analysis were performed, including protraction of growth curve, HE staining, immunocytochemical staining and scanning electron microscope observation. The tumorigenicity was assessed by colony formation and transplanting the cells into nude mice.

RESULTSHuman oral epithelial cells transfected with HPVE6/E7 has been in culture for over 18 months. The cell line was named HIO615. Western blot analysis showed HIO615 expressed HPV16 E6 and E7 protein. HIOC were positive for cytokeratin, tonofibril and desmosome as observed by scanning electron microscope. The number of large colonies of dense multilayer cells was low (0.77%). No tumor developed in nude mice injected subcutaneously with HIOEC.

CONCLUSIONA human immortalized oral epithelial cell line induced by HPV16E6 and E7 has been successfully established.

Animals ; Cell Line, Transformed ; Cell Transformation, Viral ; Humans ; Mice ; Mice, Inbred BALB C ; Mouth Mucosa ; cytology ; ultrastructure ; virology ; Oncogene Proteins, Viral ; genetics ; Papillomavirus E7 Proteins ; Repressor Proteins

OBJECTIVETo investigate the expression of E-cadherin, vimentin, β-catenin and transforming growth factor-β1 (TGF-β1) in oral squamous cell carcinomas (OSCC).

METHODSEighty-nine cases of OSCC and 20 cases of normal oral mucosa were collected. Then the 89 cases of OSCC were classified as grade I, II, III. The semiquantitative method was used to calculated the positive intensity and positive rate. The relationship between the OSCC differentiation and the four biomarkers was analyzed.

RESULTSThe median of E-cadherin was 9.00 in the normal tissue, 9.00, 6.00 and 6.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-4.211, P=0.000). The median of vimentin was 0.00 in the normal tissue, 0.00, 0.00 and 4.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-3.675, P=0.000). The median of β-catenin was 9.00 in the normal tissue, 3.00, 4.00 and 3.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-6.300, respectively. There was significant difference between the normal group and OSCC group (Z=-3.329, P=0.000). E-cadherin expression was positively correlated to β-catenin expression (r=0.327, P=0.002), negtively correlated to vimentin expression (r=-0.386, P=0.001) and positively correlated to TGF-β1 expression (r=-0.304, P=0.004). Vimentin expression was positively correlated to TGF-β1 expression (r=0.401, P=0.000).

CONCLUSIONSE-cadherin and β-catenin in OSCC had a down-regulated expression, while the vimentin has an up-regulated expression.

Cadherins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Humans ; Mouth Mucosa ; cytology ; metabolism ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Transforming Growth Factors ; metabolism ; Vimentin ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo study the chondrocyte apoptosis in rabbit temporomandibular joint after anterior disc displacement.

METHODSExperimental anterior disc displacement was induced surgically in 20 Japanese rabbits without opening their temporomandibular joint bursas. Histopathologic and apoptotic (TUNEL) analysis was used to evaluate the changes in articular cartilage, disc and synovium.

RESULTSCondyle chondrocyte showed apoptosis most obviously at 1 or 2 weeks after surgery, and apoptotic cells concentrated in proliferative zone and hypertrophic zone. 4 or 6 weeks after surgery, the joint went into a remodelling period.

CONCLUSIONSChondrocyte apoptosis in temporomandibular joint will be activated after anterior disc displacement, which initiates the remodelling in temporomandibular joint.

Animals ; Apoptosis ; Chondrocytes ; pathology ; In Situ Nick-End Labeling ; Joint Dislocations ; physiopathology ; Rabbits ; Temporomandibular Joint ; physiopathology ; Temporomandibular Joint Disc ; physiopathology

BACKGROUNDTo determine the long-term efficacy and persistence of Chinese infants after receiving only active plasma-derived hepatitis B vaccine, and to evaluate if providing booster vaccination after basic hepatitis B immunization is necessary.

METHODSInfants who were born in 1986-1988 in four demonstrative hepatitis B immunization trial areas of Hunan, Guangxi, Hebei and Shanghai after receiving only active plasma-derived hepatitis B vaccination, had been randomly followed up for 15 years. HBsAg,anti-HBs and anti-HBc in 21 680 person-times were tested using commercial SPRIA kits.

RESULTSPrevalence of HBV carriers was less than 1.66% among all children vaccinated with only active plasma-derived hepatitis B vaccine in 4 clinical trial areas. Prevalence of HBsAg did not increase with years after vaccination,90%(95% Cl:83.1%-97.2%) effectiveness of hepatitis B vaccine persisted for 15 years in preventing chronic HBV infection. Carriage, HBV infection and efficacy were not different among all age groups (P>0.05). Seroprotection rate (anti-HBs?10 mIU/ml) and quantity of anti-HBs were significantly decreased with years after vaccination. Seroprotection rates of anti-HBs were 40%-50% and 30%-42% during the 9th-10th year and the 13th-14th ear of vaccination, respectively. Titer of anti-HBs declined?by 90% after 14 years.

CONCLUSIONSThese results showed that long-term efficacy of only active plasma-derived hepatitis B vaccination, which was not affected by decline in seroprotection rate and titer of anti-HBs. For children and adults whose immune status is normal, booster doses of vaccine are not recommended.

Adolescent ; Child ; Child, Preschool ; Follow-Up Studies ; Hepatitis B ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B Vaccines ; immunology ; Humans ; Infant ; Infant, Newborn ; Vaccination