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Heo, Si-Hyun,Han, Kyu-Boem,Lee, Young-Jun,Kim, Ji-Hyun,Yoon, Kwang-Ho,Han, Man-Deuk,Shin, Kil-Sang,Kim, Wan-Jong The Korean Society for Integrative Biology 2012 Animal cells and systems Vol.16 No.3
Burns are one of the most devastating forms of trauma and wound healing is a complex and multicellular process, which is executed and regulated by signaling networks involving numerous growth factors, cytokines, and chemokines. Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) was specifically produced from rice cell culture through use of a recombinant technique in our laboratory. The effect of rhGM-CSF on promotion of deep second-degree burn wound healing on the back skin of a hamster model was evaluated through a randomized and double-blind trial. As macroscopic results, hamster skins of the experimental groups showed earlier recovery by new epidermis than the control groups. Immunohistochemical reactions of proliferating cell nuclear antigen and transforming growth factor-b1, which are indicators of cell proliferation, were more active in the experimental group, compared with the control group. On electron microscopy, basal cells in the epidermis of the experimental group showed oval nuclei, prominent nucleoli, numerous mitochondria and abundant free ribosomes. In addition, fibroblasts contained well-developed rough endoplasmic reticulum with dilated cisternae. Bundles of collagen fibrils filled the extracellular spaces. Particularly, ultrastructural features indicating active metabolism for regeneration of injured skin at 15 days after burn injury, including abundant euchromatin, plentiful free ribosomes, and numerous mitochondria, were observed. These findings suggest that use of rhGM-CSF could result in accelerated deep second-degree burn wound healing in animal models.
( Mi Jin Lee ),( Kyu Boem Han ),( Eung Soo Kim ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.1
A cyclic undecapeptide-family natural product, cyclosporin A (CyA), which is one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex in a filamentous fungal strain named Tolypocladium niveum. Previously, structural modifications of cyclosporins such as a regionspecific hydroxylation at the 4th N-methyl leucine in a rare actinomycetes called Sebekia benihana were reported to lead to dramatic changes in their bioactive spectra. However, the reason behind this change could not be determined since a system to genetically manipulate S. benihana has not yet been developed. To address this limitation, in this study, we utilized the most commonly practiced gene manipulation techniques including conjugation-based foreign gene transfer-and-expression as well as targeted gene disruption to genetically manipulate S. benihana. Using these optimized genetic manipulation systems, a putative cytochrome P450 hydroxylase (CYP) gene named CYP506, which is involved in CyA hydroxylation in S. benihana, was specifically disrupted and genetically complemented. The S. benihana?CYP506 exhibited a significantly reduced CyA hydroxylation yield as well as considerable yield restoration by functional complementation of the S. benihana CYP506 gene, suggesting that the genetically manipulated S. benihana CYP mutant strains may serve as a more efficient bioconversion host for various valuable metabolites including CyA.
Kim, Ji-Hyun,Han, Kyu-Boem,Choi, Yong-Soo,Lee, Young-Jun,Yoon, Kwang-Ho,Han, Man-Deuk,Kim, Wan-Jong Korean Society of Microscopy 2012 Applied microscopy Vol.42 No.3
Human placenta extract (hPE) has therapeutic potential against certain diseases such as burn injury, liver cirrhosis and chronic wound through stimulating tissue repair processes. However, the effects of hPE on liver regeneration in animals are unknown. This study investigated the effect of hPE on the expression of proliferating cell nuclear antigen (PCNA) during liver regeneration induced by partial hepatectomy (PH) in rats. The activities of AST, ALT and ALP increased during a few days after PH. A high level of ALP was particularly seen at day 3 in the control group. All the levels of experimental groups were normalized by day 5 after PH. On immunohistochemistry, the expression of PCNA increased at the early days, showed a peak at day 3 after PH. The PCNA staining was more obvious in the experimental group over the whole period. By western blotting, PCNA seemed to be more strongly expressed in the hPE injected group in the early stage and fell to almost undetectable levels at day 7. On immunocytochemical observations, the number of PCNA-gold particles in the nuclei at day 1 of the hPE treated groups was more than those of the untreated groups. The results suggest that hPE could accelerate liver regeneration induced by PH involving the expression of PCNA in rats.