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Yasuyuki Aoyagi,Yasushi Saito,Masayuki Kuroda,Sakiyo Asada,Hideaki Bujo,Shigeaki Tanaka,Shunichi Konno,Masami Tanio,Itsuko Ishii,Masayuki Aso 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.3
The development of clinically applicable scaffolds is important for the application of cell transplantation in various human diseases. The aims of this study are to evaluate fibrin glue in a novel protein replacement therapy using proliferative adipocytes and to develop a mouse model system to monitor the delivery of the transgene product into the blood and the fate of the transduced cells after transplantation. Proliferative adipocytes from mouse adipose tissue were transduced by a retroviral vector harboring the human lecithin-cholesterol acyltransferase (lcat) gene, and were subcutaneously transplanted into mice combined with fibrin glue. The lcat gene transduction efficiency and the subsequent secretion of the product in mouse adipocytes were enhanced using a protamine concentration of 500 μg/ml. Adipogenesis induction did not significantly affect the lcat gene-transduced cell survival after transplantation. Immunohistochemistry showed the ectopic enzyme production to persist for 28 days in the subcutaneously transplanted genetransduced adipocytes. The increased viability of transplanted cells with fibrin glue was accompanied with the decrease in apoptotic cell death. The immunodetectable serum LCAT levels in mice implanted with the fibrin glue were comparable with those observed in mice implanted with Matrigel, indicating that the transplanted lcat gene-transduced adipocytes survived and functioned in the transplanted spaces with fibrin glue as well as with Matrigel for 28 days. Thus,this in vivo system using fibrin is expected to serve as a good model to further improve the transplanted cell/scaffold conditions for the stable and durable cell-based replacement of defective proteins in patients with LCAT deficiency.
차재은(Jae-Eun Cha),사이토(Yasushi Saito) 한국가시화정보학회 2011 한국가시화정보학회지 Vol.9 No.4
The flow-field of a liquid-metal system is very important for the safety analysis and the design of the steam generator of liquid-metal fast breeder reactor. Dynamic neutron radiography (DNR) is suitable for a visualization and measurement of a liquid metal flow and a two-phase flow in a metallic duct. However, the three dimensional DNR techniques is not enough to obtain the velocity information in the wide channel up to now. In this research, a high speed DNR technique was applied to visualize the heavy liquid-metal flow field in the narrow channel with the HANARO-beam facility. The images were taken with a high frame-rate neutron radiography at 250 fps and analyzed with a Particle Image Velocime-try(PIV) method. The images were compared with the results of the commercial CFX code to study the feasibility of DNR technique for the measuring the heavy liquid-metal flow field. The PIV images could discern the turbulent vortex flow in the two-dimensional narrow channel.
Yoshitaka Fukaya,Yasuyuki Aoyagi,Sakiyo Asada,Yoshitaka Kubota,Yoshitaka Okamoto,Toshinori Nakayama,Yasushi Saito,Kaneshige Satoh,Masayuki Kuroda,Hideaki Bujo 생화학분자생물학회 2012 Experimental and molecular medicine Vol.44 No.5
Auto-transplantation of adipose tissue is commonly used for the treatment of tissue defects in plastic surgery. The survival of the transplanted adipose tissue is not always constant, and one of reasons is the accelerated apoptosis of the implanted preadipocytes. We have recently established highly homogeneous preadipocytes, named ccdPAs. The aim of the current study was to evaluate the regulation of the potency of platelet-rich plasma (PRP) on the apoptosis of ccdPAs in vitro. PRP stimulated the proliferation of the preadipocytes in a dose-dependent manner, and the stimulatory activity of 2% PRP was significantly higher than that of 2% FBS or 2% platelet-poor plasma (PPP). The presence of 2% PRP significantly inhibited serum starvation-or TNF-α/cycloheximide-induced apoptosis in comparison to 2% FBS or 2% PPP. DAPK1 and Bcl-2-interacting mediator of cell death (BIM) mRNAs were reduced in the preadipocytes cultured with 2% PRP in comparison to those cultured in 2% FBS. The gene expression levels were significantly higher in cells cultured without serum in comparison to cells cultured with 2% FBS, and the levels in the cells with 2% PRP were reduced to 5-10% of those in the cells without serum. These results indicated that ccdPAs exhibit anti-apoptotic activities, in addition to increased proliferation,when cultured in 2% PRP in comparison to the same concentration of FBS, and that this was accompanied with reduced levels of DAPK1 and BIM mRNA expression in in vitro culture. PRP may improve the outcome of transplantation of adipose tissue by enhancing the anti-apoptotic activities of the implanted preadipocytes.
Asako Nogami,Masato Yoneda,Michihiro Iwaki,Takashi Kobayashi,Yasushi Honda,Yuji Ogawa,Kento Imajo,Satoru Saito,Atsushi Nakajima 대한간학회 2023 Clinical and Molecular Hepatology(대한간학회지) Vol.29 No.-
Non-alcoholic fatty liver disease is currently the most common chronic liver disease, affecting up to 25% of the global population. Simple fatty liver, in which fat is deposited in the liver without fibrosis, has been regarded as a benign disease in the past, but it is now known to be prognostic. In the future, more emphasis should be placed on the quantification of liver fat. Traditionally, fatty liver has been assessed by histological evaluation, which requires an invasive examination; however, technological innovations have made it possible to evaluate fatty liver by non-invasive imaging methods, such as ultrasonography, computed tomography, and magnetic resonance imaging. In addition, quantitative as well as qualitative measurements for the detection of fatty liver have become available. In this review, we summarize the currently used qualitative evaluations of fatty liver and discuss quantitative evaluations that are expected to further develop in the future.
Oral Administration of Phosphorylated Dextran Regulates Immune Response in Ovalbumin-Immunized Mice
Nagasawa, Chiho,Nishimura-Uemura, Junko,Tohno, Masanori,Shimosato, Takeshi,Kawai, Yasushi,Ikegami, Shuji,Oda, Munehiro,Saito, Tadao,Kitazawa, Haruki Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.1
Phosphorylated dextran (P-Dex) is an acidic polysaccharide that functions as an immune adjuvant. P-Dex is known to regulate immune response by maintaining a balance between Th1 and Th2 cells in vitro, and thus may also be important in the control of allergic reactions. In the current study, we report the optimum conditions required for the efficient phosphorylation of dextran without toxicity. We found that when dextran was heated at 160${^{\circ}C}$ for 24 h in phosphate buffer (pH 5.0), the resulting P-Dex demonstrated the highest phosphorus content (6.8%). We also report that P-Dex enhances mitogenic activity in mouse splenocytes and induces expression of CD69 and CD86 on the surface of B cells and dendritic cells (DC) in vitro. Oral administration of P-Dex to ovalubmin (OVA)-immunized mice was found to reduce antigen-induced cell proliferation and suppress the expression of CD86 on Th2-inducing DC via exogenous OVA stimulation. P-Dex was also found to increase IL-10 expression in the splenocytes of treated mice. These findings suggest that oral administration of P-Dex increases immunological tolerance and improves the specificity of immunological response to specific antigens.