ABSTRACT
Objective To explore the labeling method of rat adipose-derived stromal cells,and observe the stem cell characteristics and the activities of EGFP-positive adipose-derived stromal cells (EGFP-ADSCs) in vitro and in vivo.Methods ADSCs were transfected for 12 h with enhanced green fluorescent protein gene (EGFP) carried by lentivirus(Lv-EGFP) vector at different value of MOI (0,5,10,25,50,100,respectively).The rate of EGFP expression and fluorescence intensity were evaluated by flow cytometric analysis and fluorescence microscopy,and cell viability was detected by MTT-test after transfection.Secondly,cells were exposed either to adipogenic medium or osteogenic medium,then stained with Oil Red O and Alizarin Red S.Cell growth was investigated on frozen longitudinal sections when EGFP-ADSCs were injected into acellular nerves to build tissue-engineered peripheral nerves repairing sciatic nerve defects in rats for 1 week in vivo.Results EGFP-positive rate and fluorescence intensity peak at 4 days after transfection.The rate of EGFP expression was 0.13％,31.09％,75.33％,92.66％,96.70％,98.38％ for MOI =0,1,5,25,50,100,respectively.The positive rate between the experimental group and control (MOI =0) existed significantly difference (P ＜ 0.05) ; the difference between MOI =1,5 groups and MOI =25,50,100 groups were also observed (P ＜ 0.05).There was no statistical difference in EGFP-positive rate and cell proliferation activity among MOI =25,50,100 groups (P ＞ 0.05).MOI =25 was chosen as best scheme to transfect ADSCs for subsequent experiments.Osteogenic and adipogenic differentiation for 20 days,orange calcium deposits,orange-red lipid droplets were seen in EGFP-ADSCs after Alizarin red and oil red O staining.At 1 week in vivo,EGFP-ADSCs evenly distributed and became fusiform on frozen longitudinal sections.Conclusion Lv-EGFP transfection does not affect the ADSCs activity and their osteogenic and adipogenic differentiation,so could be as a tracing method for ADSCs-tissue-engineered peripheral nerves repairing nerve defects.
ABSTRACT
Objective To explore a method of preparing the acellular tendons of human with chemical approaches.Methods From April 2011 to June 2012,several treatments were performed on human tendons using 1.00％ tri(n-butyl) Phosphate (TnBP) combinded with Triton X-100 at the concentration of 0,0.25％,0.50％,1.00％ respectively.Specimens were examined using histological observation,scanning electron microscopy,biomechanical testing and hydroxyproline quantitation.The results were then compared with fresh tendons of control group,so as to value the characteristics of decellularized human tendon scaffold.Results Acellular human tendons had glossy surface,intact aponeurotic membrane and satisfactory flexibility.A small number of disrupted cells remained in the 1.00％ TnBP + 0％ Triton X-100 treated tissue,while other three experimental groups successfully eliminate all cells.Intact and regular collagen architecture was retained in 1.00％ TnBP +0.25％ Triton X-100 treated tissue.1.00％ TnBP + 0.50％ Triton X-100 and 1.00％ TnBP + 1.00％ Triton X-100 treated tissue were nearly identical to 1.00％ TnBP +0.25％ Triton X-100 treated tissue,but the interval of collagen was slightly wider than the control group,the maximum load (385.22 ± 80.32N,398.22 ± 127.20N),ultimate tensile strength(46.69 ± 16.30Mpa,46.20 ±5.52Mpa) and hydroxyproline content(0.282663 ± 0.0110109 μg/mg,0.279355 ± 0.0102129 μg/mg) were statistically lower (P ＜ 0.05) compared with those of the control group,maximum load(533.28 ± 135.77N),ultimate tensile strength (65.56 ± 14.40Mpa) and hydroxyproline content (0.292882 ± 0.0100988 μg/mg) respectively.Conclusion The decellularization treatment with 1.00％ TnBP + 0.25％ Triton X-100 could be optimized for preparing acellular human tendons.
ABSTRACT
Objective To optimize microwave-assisted extraction of polyphenols from Enteromorpha prolifra.Methods Based on single-factor tests,an efficient microwave-assisted extraction(MAE)technique was developed to extract bioactive polyphenols from E.prolifra through orthogonal L16(4)5 test.Results The highest yield(0.923±0.013)mg/g was obtained when microwave power,solvent to raw material ratio,irradiation time,ethanol concentration,and extraction cycles were 500 W,25 mL/g,25 min,40%,and 3,respectively,which was higher than that of Soxhlet extraction with methanol for 6 h,ultrasound-assisted extraction with 40% ethanol for 1 h twice and heat reflux extraction with 40%ethanol for 2 h twice.Conclusion This finding indicates that MAE is a superior technique for the extraction of polyphenols due to less impurity,higher time efficiency and yield.
ABSTRACT
ObjectiveTo study the expression of transforming growth factor beta-1 (TGF-?1),platelet endothelial cell adhesion molecule-1 (PECAM-1,CD31) in prostate cancer tissues and their correlation with prostate specific antigen (PSA) values and TNM staging.MethodsUsing immunohistochemical method 36 prostate cancer specimens were tested for TGF-?1 expression, and using CD31 for marking vascular endothelial cells,the tumor microvascular density (MVD) was counted. Twelve normal tissue specimens were taken from the non-tumorous tissues adjacent to the prostate cancer as controls. The correlation of TGF-?1 expression and MVD with PSA values,TNM staging,pathologic grading and bone metastasis was analyzed in combination with the clinical data.ResultsThe positive expression rate of TGF-?1 in prostate cancer was 88.89%(32/36),while it was 16.67%(2/12)in controls, showing statistically significant difference between them ( P 20 ng/ml, MVD was (81.5?12.2) mm 2 ( P 20 ng/ml,23 cases had the TGF-?1 expression rate of 100% ( P