Objective To evaluate urinary vascular endothelial growth factor (VEGF) in the diagnosis of bladder cancer. Methods VEGF in urine was measured by enzyme linked immunosorbent assay in 33 cases of transitional cell bladder carcinoma,10 benign urological conditions and in 10 normal individuals. Results The mean urinary VEGF level in 33 patients with bladder cancer was ( 309.8 ? 86.6 )ng/g Cr, significantly higher than in the 20 controls which was (107.6?35.4)ng/g Cr ( P

[Objective]To evaluate the radiographic and clinical results of locking compression plate and interlocking intramedullary nail in the treatment of distal tibial shaft fracture(4 to 11cm proximal to tibiotalar joint).[Method]A total of 65 cases of distal tibia shaft fractures were retrospectively reviewed from 2003 to 2007.Thirty-seven cases were treated with locking compression plate as group A,and 28 cases were treated with interlocking intramedullary nail as group B.According to AO classification system,32 cases were type A,12 were type B,21 were type C.[Result]All cases were followed up for 12-24 months(mean,16.5 months).Five cases were found delayed or nonunion,with 1 case in group A and 4 cases in group B.One case in group A and 2 cases in group B were found developed deep infection.Two cases in group A and 8 cases in group B received second operation.The good-to-excellent rates of ankle joint function were 91.9% in group A and 71.4% in group B.The good-to-excellent rates of knee joint function were 97.3% in group A and 78.6% in group B.[Conclusion]Distal tibial shaft fracture could be treated successfully with either percutaneous plates or intramedullary nails.Percutaneous locking plate is better than intramedullary nail in aspect of prevention of delayed union,malunion and second operation.

[Objective] To investigate the effectivity and reliability of individualized method to localize the entrance point of the lumbar pedicle screws by vertebral plate borderline.[Methods]Four vertebra specimen were taken the X-ray film of anterior-posterior views as the center of L2.On the X-ray film,the distance between the centre-point of the pedicle to the lateral and superior margins of the vertebral plate were measured,after applied barium at the centre-point of the pedicle,the specimen were taken the X-ray film once more,the relation of the barium to the centre-point of the pedicle was measured;the distance between the centre-point of the pedicle to the lateral and superior margins of the vertebral plate were measured on the normal lumbar X-ray film in 107 cases,the data in different sex and different projection 's center were compared;the method were applied to 40 patients of lumbar pedicle internal fixation,the information in hospital were reviewed to 30 patients of the lumbar pedicle screw localization by transverse process and transverse process.[Results]All(except L5)of lumbar plate borderline were displayed clear in the experiment,accurate rate of setpoint to be 92.5%.The principal X-ray affected scarely to the relation of the pedicle with the plate borderline of nearby two segment,but affected obviously far from three segment on the normal lumbar X-ray film;the accurate rate of one time 's setpoint in clinical application was 96%,average operation 's time being 96 minute,average lose blood being 212 ml,transverse articular process 's group showed significant difference.[Conclusion]Individualized localization of the lumbar pedicle screw entrance points by vertebral plate borderline is one of feasible method,displayed less trauma and accurate rate highly in the clinical application.

Objective To investigate the stability of lumbar single?level dynamicly fixed by socket track pedicle screws after decompression. Methods Socket track pedicle screws are made of titanium?alloy, swing like universal screw within the scope of 18° in the sagittal plane and 8° in the horizontal plane. Motion constraints were due to the hole which the screw goes through. Six pig spines (L1-5) were tested by applying a pure moment of 4.018 Nm in 3 directions of loading. The range of motion (ROM) of the following groups underwent different surgical procedures was measured:(1) intact;(2) dynamic fixation 1 (decompres?sion by total laminectomy and instrumentation with socket track pedicle screws); (3) dynamic fixation 2 (decompression by total laminectomy and bilateral facetectomy while instrumentation with socket track pedicle screws);( 4) instability resulted from decom?pression (decompression by total laminectomy plus bilateral facetectomy without any rod);(5) hybrid fixation (decompression by to?tal laminectomy and bilateral facetectomy while instrumentation with 2 socket track pedicle screws connected with 2 unilateral pedicle screws ). Results Compared with the intact specimens (in flexion, extension, lateral bending and rotation, corresponding angular displacements were 1.84°±0.75°, 1.55°±0.34°, 2.2°±1.07°, 2.06°±0.76° respectively), the ROM of segment L3, 4 of each group were significantly restricted except for axial rotation. In flexion, extension and lateral bending, the ROM was reduced to 0.71°±0.39°, 0.46°±0.18°, 0.85°±0.66° in dynamic fixation 1 group;0.79°±0.43°, 0.71°±0.20°, 1.17°±0.48° in dynamic fixation 2 group and 0.63°±0.50°, 0.37°±0.19°, 0.54°±0.60° in hybrid fixation group. The rotation ROM of dynamic fixation 1 group and hy?brid fixation group were decreased to 1.50°±0.64° and 1.22°±0.36°, whereas the ROM of dynamic fixation 2 had a non?significant reduction to 1.85°±0.67°. The extension and rotation ROM of dynamic fixation 2 increased significantly comparing to dynamic fixa?tion 1 group; the extension, lateral bending and axial rotation ROM of hybrid fixation group were reduced comparing to the state of dynamic fixation 2 group. Conclusion The dynamic instrumentation of single?level lumbar spine with socket track pedicle screws could offer stability in all directions. The flexion, extension, lateral bending and rotation ROM of spine with dynamic instru?mentation after total laminectomy and bilateral facetectomy could be reduced.

Objective:To produce and identify antiCK20 monoclonal antibody.Methods:Lymphocytes from the spleen of mice being immunonized by CK20 antigen were fused with the myeloma cell line(SP2/0) using PEG4000.Hybridodma cells were established by selective growth of the fusion cells in the HT medium,and the presence of antiCK20 antibody was screened by inderect ELISA,and the clonality was achieved by limiting dilution.We have incubated cloning cells into mouse abdominal cavity to produce ascitic fluid contained monoclonal antibody.Chromatography with SPA-Sepharose CL-4B affinity column were emploied to isolate the monoclonal antibody from ascitic fluid.Finally,the antibody were tested the activities and sentivities,isoforms and titer through Western blot,two directions immuning diffusion of agar and ELISA.Results:Only one hybridoma cell line,secreating McAb against CK20,had been established.The modal number of chromosome is 101(99-103).The results of identifications showed that the antibodies kept high activitis and sensitivitis in detecting sample.The titer of ascitic fluid and the McAb purified are 1∶10~6 equally.The immunoglobulin of the McAb is classified as IgG1.Conclusion:AntiCK20 monoclonal antibody have been produced succesfully with high sensitive and active and was named L20031030.

Objective To explore the outcomes of surgery of hip fracture in elderly pafienta aged equal or greater than 80 years old.Method The data of totally 132 patients with hip fracture aged equal or greater than 80 years old were treated with surgery between January 2004 and December 2008 was analyzed retrospectively Results Of 132 cases,68 cases were femoral neck fracture,while 64 cases were intertroehanteric fracture.The average in.hospital time waft(17.2±4.6)d,and 2 cases died of cerebral embolism and respiratory dysfunction in perioperative period respectively.One hundred and eight cases were followed up for 6 months to 5 yeats,with an average of 3.2 years.According to Harris hip scores,the average score of femoral neck fracture at 1 year and at 2 years postoperatively were(77.6±10.2),(77.3±9.3)scores,respectively;while intertrochantefic fracture at 1 year and at 2 years postoperatively were(84.6±9.8),(83.9±10.1)scores,respectively.Conclusions Aged patients with hip fracture should receivesurgical treatment as early as possible.The reasonable perioperative treatment and operative method are the keys to gain good outcomes.

Aim To establish in vitro blood-brain barrier (BBB) model with characteristics of simulation of in vivo BBB by primi-tive co-culture of brain-microvessel endothelial cells (BMECs) with brain-microvessel pericytes (BMPC)and astrocytes (AS). Methods BMECs,BMPC and AS from SD rats were primitively isolated,purified and cultured,and then primitive culture cells were identified by cellular morphological and immunocytochemi-cal staining methods.Five types of in vitro BBB models were es-tablished by using Millicell culture insert (pore diameter 0.4μm)and their barrier functions were evaluated by detection of transendothelial electrical resistance (TEER),permeability of sodium fluorescent (Na-FLU ),expression of alkaline phospha-tase (AKP)and γ-glutamyl transpeptidase (γ-GT1 ),and simi-larity of permeation amount for positive drugs in vitro and in vivo BBB conditions.Results Primitive culture of BMECs presented typical pebbles-like structure,BMPC presented larger soma with branching property,AS presented slender synapse and shallower cytoplasm.Moreover,immunocytochemical staining results iden-tified primitive cells were targeted cells.TEER value for co-cul-ture of BMECs,BMPC and AS reached (478 ±25 )Ω· cm2 , permeability coefficients (Papp )value of Na-FLU was [(8.23 ± 0.78) ×10 -6 ]cm·s-1 ,expression of AKP and γ-GT1 were (6.90 ±0.27 )King unit · g-1 Pro and (4.39 ±0.32 )μg · g-1 Pro respectively.Moreover,good correlation could be found in Papp for positive controls in vitro and in vivo BBB models (R2=0.92).Conclusion The established in vitro BBB model by using primitive co-culture of BMECs with BMPC and AS posses-ses in vivo BBB properties in cell morphology,structures and barrier functions,and can be used as a powerful tool for studying physiology,pathology of BBB and screening candidate com-pounds.

Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .

Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.

Objective To investigate the effects of quercetin on glial scar formation and axonal regeneration after spinal cord injury (SCI) and its association with the p38 mitogen activated protein kinase (MAPK) signal pathway.Methods 128 female Sprague-Dawley (SD) rats were randomly divided into a control group (SCI + saline),an intervention group (SCI + quercetin + anisomycin),a treatment group (SCI + quercetin) and a sham-operation group (n =32).Basso Beattie Bresnahan (BBB) assessment and footprint analysis of the hind limb were performed on days 1,3,7,14,21 and 28 postoperation in each group.The expression levels of p38MAPK,phosphorylation p38MAPK,glial fibrillary acidic protein (GFAP) and neurofilament protein-200 (NF-200) were detected by Western blot.The numbers of GFAP and NF-200 positive staining cells in the injured spinal cord in each group were detected by immunohistochemistry.Results The BBB scores in the treatment group were significantly higher than in the intervention and control groups at each time point after SCI except on day 3 postoperation (P ＜ 0.05).The expression levels of phosphorylation p38MAPK protein in each SCI group were significantly higher than in the sham-operation group on days 3 and 7 postoperation (P ＜ 0.05).The expression levels of phosphorylation p38MAPK protein in the treatment group were significantly lower than in the control and intervention groups on days 3,7 and 14 postoperation (P ＜ 0.05),but there was no significant difference on day 28 postoperation among all the groups (P ＞ 0.05).The numbers of NF-200 and GFAP positive staining cells were significantly greater than in the sham-operation group at each time point postoperation (P ＜ 0.05);the NF-200 positive staining cells in the treatment group were significantly increased in comparison with the control and intervention groups (P ＜ 0.05);the GFAP positive staining cells in the treatment group were significandy fewer than in the control and intervention groups on days 7,14 and 28 postoperation (P ＜ 0.05).Conclusions Quercetin may have protective effects against acute SCI by decreasing glial scar formation,increasing axonal regeneration and promoting recovery of locomotor and nerve function in rats.The effects may be correlated with inhibition of the p38MAPK signal pathway.