The culture method for porcine pulmonary artery smooth muscle cells (PASMCs) established previously in our laboratory was modified as follows: (1) PASMC mixed medium (PASMCMM) containing 400 ml/L PASMC conditioned medium and 100 ml/L fetal bovine serum (FBS) was substituted for M199 medium containing 100 ml/L FBS; (2) When the PASMC was confluent by 90 %, the subculture was done at the time between the phase of logarithmic growth and stagnation phase of the cells; (3) The method of subculture: The cells were dispersed with 5 to 8 drops of a mixed digestive solution per flash, then M199 medium containing 100 ml/L newborn calf serum was added to terminate the digestion. The cells attached to the substrate of the flashes. On the 2nd day, the media were changed by PASMCMM. The cultured PASMCs by this method were characterized by rapid growth and good dispersion, etc.. The doubling time of the cells (45.6 h) by this method was 11.2 h shorter than that (56.8 h) by the previously established method in our laboratory.The modified culture method for PASMC was more simple, practical and effective, so it can be used as an effecrive method for rapidly harvesting large quantities of PASMCs.

AIM: To investigate the regulatory effect of the adenosine triphosphate binding cassette transporter A1 (ABCA1) on apolipoprotein E secretion from human THP1 macrophages.METHODS: Differentiation of THP1 macrophages from monocytes was stimulated with phorbol 12-myristate 13-acetate. The macrophages then were incubated with factors which regulate ABCA1 expression. After periods of incubation, apo E secreted in the medium and synthesized in the cell was determined with ELISA, and apo E mRNA espression was detected with Northern blot.RESULTS: An increase in apo E secretion from THP1 macrophages was observed by 8 h of incubation with 8-Br-cAMP, an activator of ABCA1 expression (P

To investigate whether apoptosis is associated with cell adhesion in bronchial epithelium, and whether it contributes to the kinetics of injury and repair of surface epithelia, this study was performed for E-cadherin expression by using immunohistochemistry technique and for apoptosis by TUNEL method. An animal model of smoking was used for this study. The results showed that epithelial cells with membrane anchored E-cadherin decreased remarkably at several time points during 6 months of exposure to smoke (P < 0.01) and then restored to normal level. This fluctuation was associated exclusively with the alteration in number of apoptotic cells (P < 0.01). There was no significant difference in activation of nuclear transcription factor NF-kappa B among groups (P > 0.05). All these suggested that apoptosis is associated with E-cadherin expression in bronchial epithelium of smoking mouse.
*Apoptosis ; Bronchi/metabolism ; Bronchi/*pathology ; Cadherins/analysis ; Cadherins/*biosynthesis ; Epithelial Cells/chemistry ; Epithelial Cells/metabolism ; Epithelial Cells/pathology ; Smoking/*adverse effects

In this paper are described the construction of biological valves poppet and the establishment of mathematical model according to cardiac valves hydrokinetics and the theory of membrane. Finite elements of biological valves leaflets connected with biological valves poppet are analyzed. The method of constructing geometric model is reasonable. CAD of biological valves poppet is realized by structured programming and parameterization.
Bioprosthesis ; Computer Simulation ; Computer-Aided Design ; Finite Element Analysis ; Heart Valve Prosthesis ; Humans ; Models, Theoretical ; Prosthesis Design ; methods

BACKGROUND:Herba epimedi, a traditional Chinese medicine, has a long time in dealing with various orthopedic disorders. Icarinwithmany biological activites is one of the most important compositions of Herba epimedi. OBJECTIVE:Toinvestigate the effects of icarin on osteogenic differentiation of mesenchymal stem cels and the underlying mechanisms. METHODS:Bone marrow mesenchymal stem cels were treated using icarin with or without osteogenic mediumin vitro. Osteogenic differentiation markers, including runt-related transcription factor 2, osteocalcin and osterix, were detected by real time-qPCR. Alizarin red staining was used to measure calcium nodes generated by osteoblasts induced frombonemarrow mesenchymal stem cels. The proximal tibia bone structure of rats fed with icarin (2 mgperday) for 5 weeks was detected and analyzed by MicroCT. RESULTS AND CONCLUSION:Icarin was able to promote the expression of genes related to osteogenic differentiation in the absence or presence of osteogenic induction. Icarin could obviously increase the quantity of calcium nodes whenmesenchymal stem celswere cultured in the osteogenic medium. The animal experiment showed that icarin improved formation of trabecular bone.

With development of medical imaging,especial the wide application of CT,MRI and so on,traditional education method can not meet the needs of the intern-clinical students.Fairly satisfying teaching effects has been achieved for the effective combination of traditional teaching methods with net-work educational method.

With development of medical imaging,traditional picture has been replaced by digital information.To apply information-based education to medical imaging and enhance the teaching quality,the article explored the teaching methods of information-based education.

Virtual reality technology and force feedback technology are novel human-machine interaction technologies. The virtual surgery simulation training system combined with these two technologies provides a new method for orthopedic surgery training, which can improve the training efficiency,thereby reducing the training costs and shortening the growth cycle of young orthopedic surgeons. In recent years, the virtual drilling bone surgery simulation technology have been researched broadly and obtained a preliminary application. In this paper, the existing research statusof virtual bone drilling operation depended on visuo-haptic techniques were studied, classified and summarized, the main content focused on three key techniques: bone modeling, drilling bone force prediction model and tactile simulation, and then analyzed the advantages and disadvantages of existing methods. Finally,some perspectives for related technology development trend of the virtual simulation bone drilling surgery in future was pointed out.

AIM: To explore the effect of pyrrolidine dithiocarbamate (PDTC), an NF-κB inhibitor, on the proliferation and apoptosis of human multiple myeloma U266 cells and its mechanisms.METHODS: The U266 cells were treated with PDTC at different concentrations (0, 25, 50, 100 and 200 μmol/L) in vitro.The growth inhibitory rate of the U266 cells was detected by CCK-8 assay and cell counting.The cell cycle of the U266 cells was determined by flow cyto-metry, and the apoptosis was examined by flow cytometry with Annexin V-FITC/PI staining.The effect of PDTC on the expression of DNA methyltransferase 1 (DNMT1) at mRNA and protein levels was measured by RT-qPCR and Western blot, respectively.The effects of PDTC on the protein levels of NF-κB (P65), DNMT1, Bcl-2, cyclin D1, cleaved caspase-3 and cleaved caspase-8 were determined by Western blot.RESULTS: The protein level of NF-κB (P65) was decreased after treatment with PDTC for 48 h or 72 h.PDTC inhibited the proliferation of U266 cells in both dose-and time-dependent manners.After treatment with PDTC for 48 h, the percentage of U266 cells in G2 phase increased compared with control group (P<0.05).PDTC induced the apoptosis of U266 cells in a dose-dependent manner.The expression of DNMT1 at mRNA and protein levels decreased (P<0.05).The results of Western blot showed that the expression of Bcl-2 in PDTC groups decreased, while the protein levels of cyclin D1, cleaved caspase-3 and cleaved caspase-8 were higher than those in control group (P<0.05).CONCLUSION: The NF-κB inhibitor PDTC inhibits the proliferation of U266 cells by inducing cell apoptosis.It may be related to the down-regulated expression of DNMT1, cell cycle arrest and activation of the apoptotic pathways.

Objective To discuss the effect of Dickkopf-1 (DKK-1) on Wnt singal pathway during the differentiation of osteoblast in vitro.Methods The osteoblasts were obtained from the new rats and cultured in vitro.The 3 passages were divided into control group and DKK-1 group.The cells were cultured in DKK-1 and normal saline for morphogical detection,ALP activity detection and osteoblasts stained at 1 st,6th,12th,21st day.The Wnt was detected by RT-PCR.Results After cultured by the DKK-1 in vitro,the ALP and mineralization of osterblasts staining were prlonged with culture time.Compared with control group,the expression of Wnt was significantly reduced at the 21st day after induction (t =0.278,P ＜ 0.05).Conclusion DKK-1 can regulate the expression of Wnt during osteoblast differentiation,suggests that Wnt may be involved in osteoblast differentiation and can affect bone remodeling process.