ABSTRACT
BACKGROUND:LIM mineralization protein-1 (LMP-1) and hypoxia-inducible factor-1α (HIF-1α) as intracelular proteins can induce osteogenic differentiation and promote angiogenesis, respectively. Therefore, their combination is of great significance for effectively inducing the osteogenic differentiation of adipose-derived stem cels. OBJECTIVE:To study the osteogenic differentiation of adipose-derived stem cels transfected by lentivirus vector carrying LMP-1 and HIF-1α. METHODS:Reverse transcription-PCR technology was employed to clone LMP-1 and HIF-1α genes, and thegenes were cloned to lentivirus vectors pLVX-EF1α-DsRed-Hyg and pLVX-EF1α-IRES2-AcGFP1 to construct main lentiviral vectors pLVX-EF1α-DsRed-Hyg-RLMP-1 and pLVX-EF1α-IRES2-AcGFP1-RHIF-1α. Then, Lenti-X 293T cels were transfected with main vectors pLVX-EF1α-DsRed-Hyg-RLMP-1 and pLVX-EF1α-IRES2-AcGFP1-RHIF-1α, packaging plasmid and coated plasmid. After that, lentiviral vectors were packaged to transfect adipose-derived stem cels from rats that were obtained by tissue explants culture and enzyme digestion methods. At 3, 7, 14 days after transfection, reverse transcription-PCR technology was adopted to detect the expression of osteogeic genes, such as bone morphogenetic protein 2, Runx-2, alkaline phosphatase, osteocalcin as wel as to detect the expression of vascular endothelial growth factor. RESULTS AND CONCLUSION:Lentiviral vectors pLVX-EF1α-DsRed-Hyg-RLMP-1 and pLVX-EF1α-IRES2-AcGFP1-RHIF-1α were effectively transfected into adipose-derived stem cels. Reverse transcription-PCR results showed that from the 7th day to the 14th day after lentivirus transfection, bone morphogenetic protein 2, Runx-2, alkaline phosphatase and osteocalcin al over-expressed. These findings indicate that the combination of LMP-1 and HIF-1α can enhance the osteogenic activity of adipose-derived stem cels.
ABSTRACT
Objective To evaluate the effect of rapid-prototyping (RP) fracture model in the teaching of extremity fracture. Methods 60 clinical medical undergraduates of the Fourth Military Medical University, who were receiving “surgery” teaching in 2012, were randomly divided into A and B groups equally by lottery. Undergraduates of Group A were taught by traditional methods while undergraduates of Group B were explained by using RP fracture model about the basic anatomy, fracture mechanism, injury mechanism of important vessels and nerves besides traditional methods. Degree of satisfaction of the undergraduates and examination were applied immediately after class. Original data were imputed into SPSS software (version 17.0) for comparison between the two groups using t-test. Results The difference of the two groups in the degree of satisfaction of the undergradu-ates, the average score of fracture mechanism, injury mechanism, clinical manifestation, key point of diagnosis, treatment principles and the total score all has statistical significance (P=0.000). Conclu-sions Students' self-initiative, learning interest and studying efficiency can be inspired by RP fracture models and their professional examination performance has also improved significantly.
ABSTRACT
BACKGROUND: There are pressure sensors in carotid-sinus, which are very sensitive to blood pressure regulated by ions and play an important role in the regulation of blood pressure. But it is yet not very clear how different ions regulate the blood pressure through pressure sensors in carotidsinus.OBJECTIVE: To observe the effect of different ions at various concentrations outside the carotid-sinus.DESIGN: Self-control experiment.SETTING: Preclinical Experiment Center, Fourth Military Medical University of Chinese PLA.MATERIALS: The experiment was accomplished in the Preclinical Experiment Center, Fourth Military Medical University of Chinese PLA from December 2000 to June 2001. Totally 18 New Zealand pure strain rabbits were provided by the Aninal Experimenting Center of Fourth Military Medical University of Chinese PLA. They were standard grade Ⅱ, of either gender and body mass was (2.0±0.2) kg.METHODS: The rabbits were divided into Na+, K+ and Ca2+ groups according to random numbers, and each group consisted of 6 rabbits. After anaesthesia, tracheal intubatton was performed on the rabbit, and bilateral carotid arteries were separated with carotid-sinus separated on one side and vessel intubatton performed in the other side for blood pressure measurement. Then various concentrations of Na+, K+ and Ca2+ ions were added outside the carotid-sinus with the pipette to make the carotid-sinus completed immersed in the ion solutions. The basal blood pressure and the peak value after ions addition were recorded respectively.MAIN OUTCOME MEASURES: The basal blood pressure and the peak value after ions addition.RESULTS: After Na+ (0.15, 1.5 mol/L) was added the blood pressure was(97±12), (83±17) mm Hg. It was decreased significantly compared with the basal value (106±14), (105±12) mm Hg (t=2.946, P ＜ 0.05). K+ (0.4 mol/L)decreased the blood pressure significantly [(106±12), (64±13) mm Hg, (t=13.496, P ＜ 0.01)], but other concentrations of K+ were not effective. Ca2+(0.07 mol/L) increased the blood pressure to (113±16) mm Hg compared with the basal value (103±12) mm Hg (t=-3.627, P ＜ 0.01).CONCLUSION: Na+, K+ and Ca2+ regulate the blood pressure by acting on the carotid-sinus directly. High concentrations of Na+ and K+ possess the effect of decreasing the blood pressure, while high concentrations of Ca2+increases it, which may be an important mechanism of blood pressure regulation.
ABSTRACT
Objective:To study heat shock proteins(HSP) and their association with Hantavirus nucleocapsid protein(HV-NP) in Vero-E6 cells infected with Hantaan76-118.Methods: The HV-NP was identified by immunocytochemical staining after infected with Hantaan76-118.The expression of HSP was detected by Western-blot and analyzed by double specific antibody sandwich ELISA.Results: Western-blot exhibited that the expressions of HSP27,HSP70 and Grp94 in the Vero-E6 cells infected with Hantaan 76-118 were significantly higher than those in the control cells(P
ABSTRACT
Objective:To prepare polyclonal antibody against human immunoglobulin G(human IgG) and to use this antibody in the clinical diagnosis of nephrosis. Methods:New Zealand rabbit was immunized subcutaneously with human IgG.Enzyme linked immunosorbent assay(ELISA) was applied to reveal the titer of the prepared polyclonal antiserum against human IgG.Antiserum was purified with affinity chromatography,and the purified antibody was confirmed for its specificity by Western blot and immunohistochemical staining.The purified antbodies which have been indentified were fused with FITC(fluorescein isothiocyanate) and then analyzed by immunohistochemical staining.Finally the fused antibody was useed in the clinical diagnosis of nephrosis. Results: The titer of the obtained antiserum was up to(1∶128 000,) double agar diffusion test showed the titer of the antibody was 1∶32.By the method of affinity chromatography,we obtained purified antibody with the purity of 85%.ELISA,double agar diffusion and immunohistochemical staining tests showed that the specificity and titer of the antibody were not decreased sharply.The purified antibody fused with FITC also kept the specificity of the primitive antibody.When the FITC fused antibody was tried in nephrosis patients,it detected human IgG effectively. Conclusion:The polyclonal antibody can specifically recognize human IgG.This purified antibody fused with FITC can be used in the diagnosis of nephrosis.