ObjectiveTo evaluate the physicochemical properties and bicompatibility of carbon-nanotubes/hydroxypatite/chitosan scafflod for bone tissue engineering. MethodsMWCNT/n-HA/CS scaffolds wre generated by solution blending and freeze-drying technology.The morphology and composition of the scaffolds were analyzed by scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy, after this, the results of which mixed CNTS in scaffolds were evaluated. The effects of MWCNT/n-HA/CS scaffolds on adherence and proliferation of rabbit bone marrow stroma cells were assessed by scaffolds surface seeding methods, and using scanning electron microscopy, MTT assay to observe their adhesion and proliferation on scaffolds.Results MWCNT/n-HA/CS scaffolds showed abundant homogeneous pores with (87.26％) porosity. 66％ fracture strength of the scafflod was improved by MWCNT,and porosity decreased by 3％. Conclusion MWCNT/n-HA/CS scaffold can be prepared with solution blending and freeze drying process, which has fair poriness, good mechanical strength and tissue compatibility and can be applied as a bone graft material.

ObjectiveTo establish a new model of biliary peffusion in vitro,and to explore the feasibility of comparative biliary studies with the model.MethodsA set of ten-text-area model of biliary peffusion was designed in vitro.Saline,glycerol and pig bile were put into this model.The outflow of the first tube was set at 8 drops/min,and these liquids were collected through the ten sample test areas and measured at 0.5 hour,1.0 hour,1.5 hours,2.0 hours,12.0 hours and 24.0 hours.Equality of liquid amount was evaluated from each sample test area.Rods containing delayed release EDTA of ten different levels were placed in the test areas.The liquid was collected and EDTA concentrations were evaluated from the ten tubes at 24.0 hours.EDTA concentrations of the same rod in the three liquids were compared.ResultsA(1) the same time point,the amounts of saline,glycerol,pig bile flowing through the ten sample test areas showed no significant difference (P ＞ 0.01 ) ; but EDTA concentrations of the same liquid at 24.0 hours were significantly different (P ＜ 0.01 ) ; and EDTA concentrations of the same rod in the three liquids were also obviously different ( P ＜ 0.01 ).ConclusionThe same liquid flow rates and the same experimental environment can be achieved.Different test results can be obtained from different test areas in different experiments.The test results of the same kind of sample change with the changes of experimental conditions,which simulates biliary duct.

Objective To observe the change of invasion and migration of the pancreatic carcinoma cell line SW1990 transfeeted with EEF1A2 gene.Methods Pancreatic carcinoma cell line SW1990 was transfected with EEF1A2 by recombinant adenovirus vector.The alteration of motility、invasion and adhesion property of SW1990 was evaluated by wound healing assay,transwell With or without Matrigel basement membrane and adhesion assay.Results Wound healing assay revealed that EEF1A2 enhanced cell motility and transwell assay with Matrigel indicated that the average numbers of transwell cells with EEFlA2 was increased from 23.25±5.23 to 65.42±8.24(P＜0.05).The adhesive rate was substantially increased in EEF1A2 transfected SW1990 cells compared with control cells.Conclusions EEF1A2 gene can promote the migration.invasion and adhesion ability of pancreatic cancer cell in vitro.It is indicated that EEF1A2 may involve in the development of human pancreatic cancer by influencing cell biological characteristics.

Objective To investigate the feasibility of Ad-hTGF-β1 transfected bone marrow mesenchymal stem cell(BMSCs) into chondrocytes differentiation and the change of TAZ mRNA. Methods Rats BMSCs were obtained and cultured by whole bone marrow method, and then the third-generation cells were seeded into cell culture plate, and divided into three groups:Ad-hTGF-β1 transfected group,Ad-EGFP transfected group and the control group. The control group was added in common medium without any treatment while the other two groups were respectively added in serum-free medium containing Ad-hTGF-β1 or that containing Ad-EGFP. Seven days later, real-time fluorescent quantitation PCR and Western blot were employed for detecting the expression of TGF-β1 ,while immunohistochemical and Western blot for the expression of collagen Ⅱ , and real-time fluorescent quantitation PCR for the expression of TAZ mRNA. Results Seven days after the transfection, real-time fluorescent quantitation PCR revealed that the average relative expression of TGF-β1 was:Ad-hTGF-β1 group 0.863, Ad-EGFP group 0.183, and the control group 0.180; The average relative expression of TAZ was:Ad-hTGF-β1 group 0.810, Ad-EGFP group 0.416, and the control group 0.366.The expression difference of TGF-β1 and TAZ were statistically significant (P ＜ 0.05). Western blot and immunohistochemical proved strong collagen Ⅱ expression in Ad-hTGF-β1 group while it was detected a little in the other two groups. Conclusion BMSCs could be successfully and stably induced into chondrocytes differentiation by Ad-hTGF-β1. Meanwhile, the mRNA of TAZ is up regulate during the differentiation,so it is suppose that TGF-β1 improve BMSCs into chondrocytes differentiation by TAZ.

Objective To investigate the expression of the ternary complex factor Net in human pancreatic carcinoma cell line BxPC3 and its effect on cell proliferation and the expression of c-fos.Methods pEGFP-Net prokaryotic expression plasmid and empty vector pEGFP were transfed into BxPC3 cens by using lipofectamine 2000,then monoclonal cell which stably expressing Net was established.Human pancreatic carcinoma cells proliferation was detected by MTT and flow cytometry.The tuRNA and protein expression of Net and c-fos in BxPC3 cells were detected by real.time PCR and Western blot.Results Net was low expressed in BxPC3 cells.After pEGFP-Net transfection,Net wag stably expressed and the expression of c-fos was inhibited,cell proliferation was also inhibited after pEGFP-Net transfection,the inhibitory rates at the 3rd, 5th,7th day was 38.81%,55.34%and 56.92%respectively,which were significantly higher than those in the empty vector group(5.09%,12.42%,8.6%,P<0.05).G_0/G_1 phase cell was(61.79±5.67)%,which were significantly higher than(45.14±3.37)%in the empty vector group(P<0.05).Conclusions The ternary complex factor Net could inhibit pancreatic carcinoma cell line BxPC3 proliferation.Its mechanism was possibly repressing expression of oncogene c-fos.

ObjectiveTo investigate the feasibility of hTGF-β1 transfected bone mrrow mesenchymal stem cell (BMSCs) combined with calcium alginate gel in three dimensional condition to construct tissue engineering cartilage.MethodsRats BMSCs were obtained and cultured by whole bone marrow method,and then the third-generation cells were seeded into cell culture plate,and were divided into 3 groups:AdhTGF-β1 transfected group, Ad-EGFP transfected group and control group. The control group was added in common medium without any treatment while the other 2 groups were respectively added in serum-free medium containing Ad-hTGF-β1 or that containing Ad-EGFP.Seven days later,real-time fluorescent quantitation PCR and Western blot were employed for detecting the expression of TGF-β1.Then,the BMSCs which successfully transfected by Ad-hTGF-β1,were continually cultured in vitro.Andthe confound of cells-calcium alginate gel,the cell density was 1.0 × l07 per ml,were prepared and cultuered in constant temperature incubator.Ten days later,examine the morphous and proliferation of cell.Last,paraffin slice of the cell-gel confound was stained by HE,toluidine blue and Masson staining,while immunohistochemical for the secretion of collagen Ⅱ.ResultsSeven days after the transfection, real-time fluorescent quantitation PCR revealed that the average relative expression of TGF-β1 was:Ad-hTGF-β1 group 0.863,and Ad-EGFP group 0.183, the control group 0.180, and the expression difference of TGF-β1 was statistically diffence(P ＜0.05). Western blot proved strong TGF-β1 expression in Ad-hTGF-β1 group while it was detected a little in the other two groups. Globose cells were observed through inverted microscope in the calcium alginate gel.MTT proved the amount of cells were not statistically diffence (P ＞ 0.05) at different time point.HE staining proved amount cartilage lacuna formation in the gel, while the secretion of cartilage matrix were proved by toluidine blue and Masson,and immunohistochemical proved the expression of collagen Ⅱ.Conclusion BMSC trnsfected by hTGF-β1 could be successfully induced to chondrocyte, as the cells morphous maintained.This three dimensional condition could preferably mimicry cell growth pattern as in vivo.

BACKGROUND:Bioactive glass, a multi-phase composite material, has good biological activity, bone conductivity and biocompatibility, but as a bone repair material it cannot be completely degraded, and has low mechanical strength that is insufficient.
OBJECTIVE:To design a kind of bioactive glasses/chitosan composite scaffold, and to investigate its physicochemical properties and cellcompatibility.
METHODS:Hydrochloric acid solution containing 2.0%chitosan was mixed withβ-glycerophosphate at a radio of 7:1 to prepare chitosan solution. Bioactive glasses of 0.5, 1.0, 1.5 g were added into the prepared chitosan solution, and the mass ratios of chitosan and bioactive glass were 2:1, 1:1, and 1:1.5 respectively. The composite materials were immersed and mineralized in simulated body fluid for 7 days.
RESULTS AND CONCLUSION:Scanning electron microscopy showed that the composite scaffold had an interconnected porous structure with the porosity of 89%and the pore size of 100-300μm;bioactive glasses dispersed in a needle shape between the chitosan scaffolds, arranged evenly, and were ful y wrapped tightly by the scaffolds. With the increase in mass of bioactive glass, the porosity of the composites decreased, but the fracture strength gradual y increased. There was a positive correlation between the composite porosity and fracture strength. X-ray diffraction and Fourier transform infrared spectroscopy confirmed that the composite scaffold appeared to have no changes in the nature of single materials, and differential scanning calorimetry analysis showed no mass loss at normal body temperature. After 3 days of mineralization, hydroxyapatite forming on the material surface gradual y grew up as a vil ous shape, and also significantly increased in number. After 7 days of mineralization, hydroxyapatite changed from a vil ous shape to a needle shape, the amount of hydroxyapatite was increased further, and many mineralized products were in a spherical shape.

BACKGROUND: A large amount of cell dies of apoptosis in secondary injury stage after spinal cord injury.OBJECTIVE: To probe into the influence of local intraspinal infusion with danshen injection on necrosis and apoptosis of spinal cell after acute spinal cord injury.DESIGN: Randomized controlled experiment was designed.SETTING: Clinical Medical Trial Center of People' s Hospital Affiliated to Yunyang Medical College.MATERIALS: Forty-four Chinese white rabbits of first grade were employed, aged varied from 4 to 5 months, of either sex, mass weighted varied from 2.0 to 2.5 kg.INTERVENTIONS: The experiment was performed in Clinical Medical Trial Center of People' s Hospital Affiliated to Yunyang Medical College from June 2002 to July 2003. Two groups were randomized, named danshen group and the control group, 22 rabbits for each. In both groups, modified Allen method was used to prepare the model of incomplete spinal cord injury. In danshen group, danshen injection was infused from subdural catheter for 4times at 0. 3 mL/kg per day of the total amount (once every 6 hours). In the control group, physiological saline of same dose was injected. The animals were sacrificed at the 8th, 24th and 72nd hours successively after injection for pathological and histomorphologic observation, peroxide dismutase and malondialdehyde determination and determination of positive cell count of B-cell lymphoma-2 (Bcl-2), the apoptosis-inhibition gene.MAIN OUTCOME MEASURES: Cell apoptotic index and cell apoptotic rate in the region of spinal cord injury.RESULTS: Forty-four rabbits entered result analysis for all. ① Results of cell apoptosis: Apoptotic index in danshen group was less remarkably than that in the control group(13.10 ± 1. 38, 20.39±2.96, 4.101, P ＜0.01); cell apoptotic rate was lower remarkably than the control group[ (9.67 ± 1.09)%,(14.68±2.81)%, t=4.072, P ＜0.01] and Bcl-2 expression was more than that in the control group[ (19. 12 ± 4.74) /mm2, ( 13.37 ± 3.68) /mm2,t = 2. 347, P ＜ 0.01 ]. ② POD content: The result in danshen group was higher thanthe control group[ (136.20 ± 13.64) NU/mL, (101.70 ± 15.24) NU/mL,t = 4. 132, P ＜ 0.01 ]. ③ Malondialdehyde content: The result in danshen group was lower than the control group[ (1.27 ± 0. 22) nmol/mL,(2.54±0.69) nmol/mL, t=4.309, P ＜0.01] . ④Degeneration and necrosis of neuron and neural fiber: The result in danshen group was milder than the control group.CONCLUSION: After local infusion with danshen(Radix Salviae Miltiorrhizae), cell apoptosis was decreased in local spinal cord injury and cell necrosis was inhibited and alleviated after acute spinal cord injury.

Objective To compare internal fixation with hallow compression screws combined vascularized bone graft(observation group) with only three hallow compression screws(control group) in young patients' Garden III and IV femoral neck fractures. Methods The patients with femoral neck fracture were treated from January, 2004 to December, 2013 were retrospectively reviewed. A total of 417 displaced femoral neck fractures in young and mid-dle-aged patients were long term followed-up. One hundred and thirty-seven patients were underwent open reduction and internal fixation with 3 hallow compression screws combined with a greater trochanter bone graft supported by the profound branch of medial circumflex femoral artery;280 patients were treated by closed reduction with 3 hallow com-pression screws. Results Patients had been followed-up for 5-12 years. At the last follow-up point, the Harris score of flap in observation group(93.68±5.12) were higher than that in control group(92.53±6.12), while it was no sta-tistical difference(P>0.05). It was 0.7％of nonunion incidence rate in the observation group, and incidence of avascular necrosis of femoral head was 6.6％, and incidence of femoral neck shortening was 8.8％. In the control group, inci-dence of avascular necrosis of femoral head was 14.6％, nonunion incidence rate was 4.6％, and incidence of femoral neck shortening was 22.5％. The differences between two groups was statistically significance( P<0.05). Conclusion The open reduction and internal fixation which is hallow compression screws in combination with a greater trochanter bone graft supported by the profound branch of medial circumflex femoral artery is an optimal treatment for young adults with Garden III and IV femoral neck fractures.

Objective To demonstrate the feasibility of oblique lumbar interbody fusion(OLIF)com-bined with 4-screw fixation for treating two-level lumbar degenerative diseases.Methods An intact finite element model of L3-S1(MO)was constructed and validated.Then,we constructed the M1 model by simulating OLIF surgery at L3/4 and L4/5 segments on the MO model.By attachment of posterior 4-screw or 6-screw fixation to the M1 model,three 4-screw fixation models(M2-M4)and one 6-screw fixation model(M5)were established.The segmental and overall range of motion(ROM)and the peak von Mises stresses of superior endplate,cage,and posterior screw-rod were investigated under each implanted condition.Results Under the motion modes of forward flexion,backward extension,bilateral(left and right)flexion,and left and right rotation,the L3/4 ROM of M2 model and L4/5 ROM of M3 model increased,while the L3/4 and L4/5 ROM of M4 and M5 models significantly decreased compared with those of M1 model.Under all motion modes,the L4 superior endplate in M2 model and the L5 superior endplate in M3 model showed the maximum peak von Mises stress,and the peak von Mises stresses of L4 and L5 superior endplates in M4 and M5 models were close.The L3/4 cage in M2 model and the L4/5 cage in M3 model showcased the largest peak von Mises stress,and the peak von Mises stresses of cages in M4 and M5 models were close.The peak stresses of internal fixation in M2-M5 models were close.Conclusion Four-screw fixa-tion can replace 6-screw fixation in the OLIF surgery for treating two-level degenerative lumbar diseases.