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In this paper,we summarize the teaching ideas,methods,and means for the course "Theory and Experiment of Molecular Biology" in which we teach students in an architecture-network-application way,exploring the education reform for the basic medical science.
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To prepare scaffolds for heart valve tissue engineering, porcine heart valves were treated with varied concentrations of trypsin for 32, 56, 80 and 104 h or followed with DNase. And then the structure of acellular valves was observed under light microscope, scanning and transmission electron microscope. Porcine endothelial cells, human endothelial cells, and canine myofibroblasts were reseeded onto the acellularized porcine heart valve scaffolds once a day for 3 days. The valves were analyzed by immunohistochemical staining and electron microscopy. Results show that all endothelial cells and the majority of interstitial cells were removed from the heart valves after digestion with trypsin for 104 h, and the collagen fiber structure remains intact, but the space between collagen fibers increased slightly. Incubation with trypsin for 80 h and then with DNase almost removed all cells, and the collagen fiber structure and the space between the fibers remain intact. After reseeding, human endothelial cells almost fully cover the valve scaffold surface as shown by H-E staining and platelet endothelial cell adhesion molecules (PECAM-1) staining. Xenogeneic porcine endothelial cells also adhered to and grew on the scaffolds. As shown by H-E staining and actin staining, canine myofibroblasts not only adhered to the surface of valve scaffold but also migrated to the inner part of matrix after one week culture. These results suggest that the digestion of porcine heart valves with trypsin combining with DNase is a suitable method to remove cells. The acellular porcine heart valve scaffolds have a quite favorable biocompatibility with human and porcine endothelial cells as well as canine myofibroblasts.
Sujets)
Animaux , Humains , Bioprothèse , Cellules cultivées , Endothélium vasculaire , Biologie cellulaire , Transplantation , Fibroblastes , Biologie cellulaire , Prothèse valvulaire cardiaque , Valves cardiaques , Biologie cellulaire , Fibres musculaires squelettiques , Biologie cellulaire , Suidae , Ingénierie tissulaireRésumé
<p><b>OBJECTIVE</b>To review and summarize literature regarding stimulatory and inhibitory signaling pathways from different types of Fc gamma receptors (FcgammaRs).</p><p><b>DATA SOURCE</b>Articles were obtained from Medline from January 1991 to April 2002.</p><p><b>STUDY SELECTION</b>Over 100 English language papers and reviews published over the last 11 years were selected.</p><p><b>RESULTS AND CONCLUSIONS</b>Stimulatory Fcgamma receptors include FcgammaRI, FcgammaRIIA, FcgammaRIIC, and FcgammaRIII A. They transduce signals through the immunoreceptor tyrosine-based activation motif (ITAM) in subunits or in the cytoplasmic domain. Inhibitory Fcgamma receptors, such as FcgammaRIIB, are single chain receptors, transducing signals through an immunoreceptor tyrosine-based inhibitory motif (ITIM) in cytoplasmic domains. Stimulatory signals include protein phosphorylation, increase in intracellular free calcium, the production of 1,4,5-triphosphate inositol (IP(3)) and diacylglycerol (DAG) mainly through the Src-family kinases, phosphoinositide 3-kinase (PI3-K) and phospholipase C (PLC). Inhibitory signaling has been implicated in the repression of the above activities as well as inhibition of B cell responses through Src homology 2-containing inositol phosphatase (SHIP).</p>
Sujets)
Animaux , Humains , Motifs d'acides aminés , Plaquettes , Physiologie , Phagocytes , Physiologie , Récepteurs du fragment Fc des IgG , Chimie , Physiologie , Transduction du signal , PhysiologieRésumé
This paper presents how to practically cultivate students' creative ability,scientific thinking and exploration ability in the molecular biology experiment course.