Objective:To culture and identify ratendo the lialprogen it or cells,and too bserve the expression of recombin anthuman vascular endothelial growth fator-165(VEGF165)in endothelial progenitor cells transfected with pcDNA3.1(+)/VEGF165 plasmid.Methods:Endothelial progenitor cells were cultured in vitro,and immunocytochemical staining was used to detect the expression of surface antigen(CD34,CD133,KDR).After transfection of VEGF165,VEGF protein in the culture supernatant of endothelial progenitor cells was detected by enzyme-linked immunosorbent assay(ELISA),and VEGF165 mRNA in endothelial progenitor cells was detected by reverse transcription polymerase chain reaction(RT-PCR).Results:The cultured cells were identified to be endothelial progenitor cells by immunocytochemical analysis.Gel electrophoresis showed that a 576 bp product was amplified by RT-PCR in VEGF165-transfected cells,while a weak expression of VEGF165 was observed in mock-transfected and non-transfected cells.The protein level of VEGF165 in the supernatant of VEGF165-transfected,mock-transfected and non-transfected cells were 175.8?10.7 pg/ml,10.5?1.6 pg/ml and 9.3?1.3 pg/ml,respectively.Conclusion:There is small amount of VEGF165 expression in endothelial progenitor cells in vitro,and the endothelial progenitor cells transfected with pcDNA3.1(+)/VEGF165 plasmid have an increased expression of VEGF165.

Objective To design a software for multiplex video-laparoscope image management. Methods The system adopts ACCESS DBMS and uses Visual C++6.0 based on MFC to develop programs on platform of Windows XP. Results The software has the function of playback and manifold images process. Conclusion The user interface is sententious and intuitive, so the system is true of the operation habits of doctors.

Objective To observe the influence of Bailong Jieyu Granules on quality of life (QOL) of patients with cancer-ralated depression. Methods A prospective and self-control clinical trial was carried out. Sixty seven patients with cancer-ralated depression were chosen and treated with Bailong Jieyu Granules. EORTC QLQ-C30 3.0 scores of patients were assessed before and after the treatment. Results After treatment of Bailong Jieyu Granules, the scores of role function, emotional function and global quality of life were increased, while the scores of fatigue, nausea/vomiting, insomnia and appetite loss were decreased, there were significant differences (P

Objective:To explore the effect of radiotherapy on the FHIT protein expression and cell apoptosis of cervical squamous carcinoma and discuss the relationship between FHIT protein expression and cell apoptosis. Methods:Expression of FHIT protein was measured by immunohistochemical method and cell apoptosis was detected by TdT-mediated dUTP terminal nick end labeling (TUNEL) staining in 50 cases of squamous cell cervical carcinoma at ⅡB-ⅢB stages before, during (Dt 10 Gy and Dt 30 Gy), and after radiotherapy. Results:Of the 50 patients, the positive rates of the expression of FHIT protein was 56% at Dt 10 Gy, 68% at Dt 30 Gy, and 84% after radiotherapy, which were significantly increased compared with that before radiotherapy (36%, P<0.05). The positive rates of cell apoptosis was 52% at Dt 10 Gy, 64% at Dt 30 Gy and 78% after radiotherapy, which were significantly elevated compared with that before radiotherapy (28%, P<0.05). In the process of radiotherapy, cell apoptosis was positively related to the expression of FHIT protein (P<0.05). Conclusion:Radiotherapy reinforces the expression of FHIT protein and induces apoptosis cocurrently. FHIT protein has regulatory effects in cell apoptosis induced by radiotherapy.

Objective To investigate the effect of interleukin-17 (IL-17) in the gastric cancer cell migration and invasion via regulating epithelial-mesenchymal transition (EMT) and its potential function.Methods (1) Human gastric cancer cell MGC-803 lines in the logarithmic growth phase were stimulated by 0, 1 ng/mL, 10 ng/mL,100 ng/mL and 1μg/mL of IL-17 for 48 hours, and the phenotypic changes were observed.The concentration of IL-17 was selected for follow-on experiments based on the most obvious phenotypic changes.Gastric cancer cell MGC-803 which were stimulated by 100 ng/mL of IL-17 and PBS for 48 hours were allocated into the experimental group and control group, respectively.(2) The expressions of E-cadherin and Vimentin mRNA in gastric cancer cells were assayed through real-time PCR (RT-PCR).(3) The relative expressions of E-cadherin and Vimentin proteins in gastric cancer cells were assayed by the Western blot.(4) The scratch test and Transwell detection were also utilized to study the migration and invasion of gastric cancer cell MGC-803 in vitro.Measurement data with normal distribution were presented as-x ± s and comparison between groups was analyzed using the t test.Results (1) There were significant phenotypic changes in the gastric cancer cell after the different concentration of IL-17 stimulated gastric cancer cell MGC-803 for 48 hours.Cells were changed from polygonal and tight junction to spindle and loosely junction with a deterioration of cell adhesion.Cell phenotypes were gradually changed as the concentration of IL-17 was changed from 0 to 100 ng/mL.Phenotypic changes were the most obvious when 100 ng/mL of IL-17 was used, but these were non-significant as the concentration of IL-17 increased to 1 μg/mL with the death and floating of some cells.(2) The relative expressions of E-cadherin mRNA and Vimentin mRNA in RT-PCR were 0.45 ±0.13 and 1.06 ±0.23 in the experimental group and 2.39 ±0.55 and 1.23±0.41 in the control group, respectively, with significant differences (t =3.811, 2.923, P ＜0.05).(3) The results of Western blot showed the relative expressions of E-cadherin and Vimentin proteins were 0.86 ± 0.17 and 1.56 ± 0.29 in the experimental group and 1.01 ± 0.12 and 0.56 ± 0.17 in the control group, respectively, with significant differences (t =3.551, 3.601, P ＜ 0.05).(4) Cell migration in the 2 groups were detected by the scratch test at 36 hours after scratch test, and the width of scratch in the experimental and control groups were (0.76 ± 0.13) mm and (0.40 ± 0.15) mm, showing a significant difference (t =3.095, P ＜ 0.05).Transwell detection showed number of transmembrane cell in the experimental and control groups were 159 ±28 and 94 ± 18, respectively, with a significant difference (t =3.307, P ＜ 0.05).Conclusion IL-17 can promote the migration and invasion of gastric cancer cells via stimulating alteration of EMT.

BACKGROUND: SSh as a Hedgehog signal protein can promote bone development, growth and remodeling.OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) combined with Shh modified nano-hydroxyapatite/collagen (nHAC) in the repair of femoral defects in rats.METHODS: Forty-eight Sprague-Dawley rats were randomly divided into four groups, and the model of femoral defects was established in these rats. At 14 days after modeling, experimental group was implanted with the BMSCs/Shh modified nHAC, scaffold group was implanted with simple nHAC, cell scaffold group was implanted with BMSCs/nHAC,and blank control group was without any implantation. At 3, 6, 9, 12 weeks after repair, X-ray examination, bone density measurement and bone biopsy in bone defect area were performed.RESULTS AND CONCLUSION: (1) X-ray examination: The Lane-Sandhu X-ray score and bone mineral density value in the experimental group at different time points after operation were significantly higher than those in the other three groups (P < 0.05). (2) Hematoxylin-eosin staining: 12 weeks after repair, a small amount of bone tissues but no bone marrow formed in the scaffold group; a small amount of bone tissues with absence of bone marrow formed in the cell scaffold group, and the residual scaffold was visible; in the experimental group, the scaffold was completely absorbed,and mature bone and medullary cavity formed with presence of bone marrow. (3) Scanning electron microscope observation: 12 weeks after repair, irregular arrangement of bone fibers and a large number of bone fossae were observed in the scaffold group; the cell scaffold group showed a large number of osteoblasts, but bone fibers still arranged irregularly; in the presence of the Haversian system, a large number of regularly arranged bone trabeculae were detective in the experimental group. These results elucidate that the Shh modified nHAC/BMSCs complex can promote the repair of bone defects.

BACKGROUND:A steady method that can isolate endothelial progenitor cells(EPCs) need to be explored.Studies show that the EPCs riches in bone marrow with strong reproductive activity,which can be easily obtained.OBJECTIVE:To evaluate the isolating culture and biological characterization of EPCs derived from rat marrow.DESIGN,TIME AND SETTING:The in vitro cytology experiment was performed at the central laboratory of the First Affiliated Hospital of Chongqing Medical University from August to December 2008.MATERIALS:Twenty SD rats,with two-week-old,were obtained from Experimental Animal Center of Chongqing Medical University.METHODS:femurs and tibias of rats were isolated under sterile conditions,and then the bone marrow cavity was washed by 4 ℃ 0.01 mol/L phosphate buffer(PBS).Mononuclear cells were collected from marrow suspension by density gradient centrifugation,followed by cultured with M199 culture medium containing 20% fetal bovine serum to obtained cell suspension.The cells were vaccinated into culture flask(coated with fibronectin) with the density of 1?109/L,and incubated at 37 ℃ in 5% CO2 atmosphere.MAIN OUTCOME MEASURES:Morphologic change of cells was observed by an invert microscope,the expression of CD133,CD34 as well as KDR was detected by immunocytochemistry,and the flow cytometry was used to analyze the growth curve.RESULTS:The adherent cells formed clusters that were found to be characterized by density packed round cells surrounded by spindle-likes cells.Adherent cells were identified positive for CD133,CD34 and KDR by immunocytochemistry on days 4,7,10 and 14.CD133-positive cells reduced after 10 days.Flow cytometry analysis indicated that the number of CD133/KDR double positive cells gradually increased to the maximum at day 10 and then declined.KDR-positive cells gradually increased up with the time.EPCs growth curve showed that cells proliferated fast on days 3-6.CONCLUSION:EPCs can be isolated and cultured by density gradient centrifugation from marrow suspension of SD rats.These cells exhibit the characterization of endothelial cells,and can differentiate into endothelial cells.

BACKGROUND: Polylactic acid copolymer bone scaffold has excellent biodegradability, and it is easy to be shaped and can promote the formation and growth of bone tissue and blood vessel.OBJECTIVE: To observe the effects of adipose-derived stem cells(ADSCs)/poly(lactic-co-glycolic acid) (PLGA) complex on the biomechanical properties after fracture healing in osteoporotic bone.METHODS: Sixty Sprague-Dawley rats were randomly divided into four groups: blank control group received no treatment; the bilateral tibial fracture model was made after 3 months of bilateral ovarian resection in model group; the bilateral tibial fracture model was made and ADSCs were implanted into the bone after 3 months of bilateral ovarian resection in cell therapy group; the bilateral tibial fracture model was made and the PLGA/ADSCs complex was implanted after 3 months of bilateral ovarian resection in combined treatment group.The bone mineral density, callus thickness, biomechanical parameters and the microstructure of the trabecular bone were detected.RESULTS AND CONCLUSION: (1) The bone density: The bone density of the model group was significantly lower than that of the blank control group (P < 0.05); the bone mineral density of the cell therapy group and the combined treatment group was higher than that of the model group (P < 0.05), but lower than that of the control group (P < 0.05); and the bone mineral density of the combination treatment group was higher than that of the cell therapy group (P < 0.05). (2)Thickness of the callus: The thickness of the callus in the cell therapy group and combined treatment group was higher than that of the model group and blank control group (P < 0.05); moreover, the thickness of the callus in the combined treatment group was higher than that of the cell therapy group (P < 0.05). (3) Biomechanical test: The failure load, stress and shear strength, elastic modulus were decreased in the model group compared with the blank control group (P < 0.05), while the shear strain increased (P < 0.05). Compared with the model group, the failure load, ultimate stress, shear strength, elastic modulus were increased in the cell therapy group and combined treatment group (P < 0.05), and the shear strain was decreased (P < 0.05). Moreover, the combined treatment group showed more changes in these biomechanical parameters (P < 0.05). (4) The trabecular bone microstructure: The model group presented with trabecular derangement, spacing increases, and even fracture and lacuna. After ADSCs or ADSCs/PLGA transplantation,the trabecular bones increased in number, thickness, and spacing, and the number of lacunae reduced. In conclusion,ADSCs combined with PLGA in the treatment of osteoporotic fracture can significantly improve the biomechanical parameters of bone tissue after healing.