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다공성 안와충전물에서 혈관생성 및 유지에 대한 혈관전구세포와 섬유소의 역할
양재욱,이호영,박세광,양영일.Jae Wook Yang. M.D.. Ph.D.. Ho Young Lee. M.D.. Sae Gwang Park. M.D.. Ph.D.. Young Il Yang. M.D.. Ph.D. 대한안과학회 2008 대한안과학회지 Vol.49 No.7
Purpose: The effects of endothelial progenitor cells (EPCs) and fibrin on fibrovascular growth into porous polyethylene orbital implants (Medpor® sheet) were investigated using stem cells. Methods: EPCs were separated from human adipose fat tissue for culture. Fluorescence-activated cell sorting (FACS) was used to identify the phenotype and to analyze the purity of EPCs cultivated from human adipose tissue. Processed Medpor® sheets were inserted in each quadrant of the subcutaneous fat layer under the dorsal surface of 20 anesthetized athymic nude mice, using sterile methods. Medpor® sheets processed with endothelial progenitor cells and fibrin were inserted into the two top quadrants, a Medpor® sheet processed with fibrin was inserted in the lower right quadrant, and an unprocessed Medpor® sheet was inserted in the lower left quadrant of each mouse. The mice were sacrificed on the seventh day. The adhesiveness and blood vessel formation were quantified by weight and the number of blood cells within the Medpor® sheets. Hematoxylin and eosin (H&E) and toluidine blue stains were used to analyze fibrovascular and cell growth within the Medpor® sheets. Results: The sheets processed with EPCs and fibrin were heavier and contained more white and red blood cells (p<0.001) than the other sheets. The sheets processed with fibrin alone were heavier (p<0.01) and contained more blood cells (p<0.001) than the unprocessed sheets. The degree of vessel formation and tissue adhesiveness was greatest in the group of Medpor® sheets processed with EPCs and fibrin. The sheets processed with fibrin only had greater tissue adhesiveness and fibrovascular proliferation than the unprocessed Medpor® sheets. Conclusions: Endothelial progenitor cells and fibrin applied to Medpor® sheets improve fibrovascular proliferation and tissue adhesiveness. When both are applied together, a synergistic effect is seen. J Korean Ophthalmol Soc 49(7):1135-1145, 2008
Random peptide library를 이용한 C형 간염바이러스 E2 단백질 세포막 수용체의 peptide mimotope 규명
이인희,백재은,설상영,석대현,박세광,최인학,Lee, In-Hee,Paik, Jae-Eun,Seol, Sang-Yong,Seog, Dae-Hyun,Park, Sae-Gwang,Choi, In-Hak 대한면역학회 2001 Immune Network Vol.1 No.1
Background: Hepatitis C virus(HCV), a family of Flaviviridae, has a host cell-derived envelope containing a positive-stranded RNA genome, and has been known as the maj or etiological agent for chronic hepatitis, hepatic cirrhosis, and hepatocellular carcinoma. There remains a need to dissect a molecular mechanism of pathogenesis for the development of therapeutic and effective preventive measure for HCV. Identification of cellular receptor is of central importance not only to understand the viral pathogenesis, but also to exploit strategies for prevention of HCV. This study was aimed at identifying peptide mimotopes inhibiting the binding of E2 protein of HCV to MOLT-4 cell. Methods: In this study, phage peptide library displaying a random peptides consisting of 7 or 12 random peptides was employed in order to pan against E2 protein. Free HCV particles were separated from the immune complex forms by immunoprecipitation using anti-human IgG antibody, and used for HCV-capture ELISA. To identify the peptides inhibiting E2-binding to MOLT-4 cells, E2 protein was subj ect to bind to MOLT-4 cells under the competition with phage peptides. Results: Several phage peptides were selected for their specific binding to E2 protein, which showed the conserved sequence of SHFWRAP from 3 different peptide sequences. They were also able to recognize the HCV particles in the sera of HCV patients captured by monoclonal antibody against E2 protein. Two of them, showing peptide sequence of HLGPWMSHWFQR and WAPPLERSSLFY respectively, were revealed to inhibit the binding of E2 protein to MOLT-4 cell efficiently in dose dependent mode. However, few membrane-associated receptor candidates were seen using Fasta3 programe for homology search with these peptides. Conclusion: Phage peptides containing HLGPWMSHWFQR and WAPPLERSSLFY respectively, showed the inhibition of E2-binding to MOLT-4 cells. However, they did not reveal any homologues to cellular receptors from GenBank database. In further study, cellular receptor could be identified through the screening of cDNA library from MOLT-4 or hepatocytes using antibodies against these peptide mimotopes.
박세광,송미경,김희선,최인학 인제대학교 2003 仁濟醫學 Vol.24 No.1
The intracellular expression of recombinant antibodies, intrabody (intracellular antibody) was developed to target and inactivate the specific intracellular molecules in a specialized compartment. There has been several lines of evidence that put the light on the treatment of cancer, viral infection, and autoimmune disease. In this study, we aimed at the construction of vector expressing human intrabody under the control of X-protein of HBV (HBX). HBX is known to up-regulate the several promoters and enhancer: tk promoter of HSV, SV40 early promoter, HIV-LTR, and enhancer 1(En 1) of HBV. First, the gene encoding HBX was PCR-amplified and cloned into pcDNA 3.1(+) vector. HBX was expressed transiently in COS-7, not in HepG2 cells. Second, each PCR-amplified promoters and enhancer was cloned into pGL-Basic luciferase construct. pGL constructs were transfected together with pX expressing HBX into COS-7 cells, among which construct employing tk promoter exhibited a higher luciferase activity than any other constructs. Third, tk promoter sequence was recloned into recombinant plasmid expressing human light chain against preSl of HBV. Recombinant antibody was expressed in COS-7 cells cotransfected with pX, not in HepG2 cells. Based on this data, new construct would be developed to express intrabody in a specific mode only in HBV-infected hepatocytes.
Antibody Phage Display 기법과 그 응용
박세광 인제대학교 백병원 2002 仁濟醫學 Vol.23 No.2
In recent years, the use of display vectors and in vitro selection technologies has transformed the way in which we generate ligands, such as antibodies and peptides, for a given target. Using this technology, we are now able to design repertoires of ligands from scratch and use the power of phage selection to select those ligands having the desired (biological) properties. With phage display, tailor-made antibodies may be synthesized and selected to acquire the desired affinity of binding and specificity for in vitro and in vivo diagnosis, or for immunotherapy of human disease. This review addresses recent progress in the construction of, and selection from phage antibody libraries, together with novel approaches for screening phage antibodies. As the quality of large na ve and synthetic antibody repertoires improves and libraries becomes more generally available, new and exciting applications are pioneered such as the identification of novel antigens using differential selection and the generation of receptor a(nta)gonists. A combination of the design and generation of millions to billions of different ligands, together with phage display for the isolation of binding ligands and with functional assays for identifying (and possibly selecting) bio-active ligands, will open even more challenging applications of this inspiring technology, and provide a powerful tool for drug and target discovery well into the next decade.
B형 간염 바이러스 감염에 대한 항바이러스 치료를 위한 새로운 표적과 분석 기법
박세광 인제대학교 백병원 2002 仁濟醫學 Vol.23 No.5
There are presently only two licensed therapies for treating liver disease caused by infection with the hepatitis B virus (HBV). These are interferon-alpha and lamivudine. Neither agent was specifically developed as an antiviral compound for treating patients infected with HBV. Both therapies are limited in the clinic by a low response rate and in the case of lamivudine, selection of drug-resistant mutants, whilst troublesome side effects limit the use of interferon-alpha. Several promising nucleoside/nucleotide analogues are undergoing clinical trials, including adefovir dipivoxil and entecavir, both of which appear to be active against lamivudine-resistant HBV. In addition to these nucleoside/nucleotide analogues, it will be important to develop new agents with different modes of action, which can be added to the antiviral cocktails that will be required to adequately suppress and hopefully eliminate HBV replication.
박인철,박세광,한 진,최병선,김희덕 大韓應急醫學會 1999 대한응급의학회지 Vol.10 No.1
Background : Scuba diving has become increasingly popular in Korea. Medical problems are common with dives, especially decompression sickness(DCS). This study was performed to obtain an useful information of hyperbaric oxygen therapy in DCS in Korea. Method : We reviewed the 62 cases of Korean divers, who were diagnosed as DCS and received recompression therapy according to U, S. Navy Standard Recompression Treatment Table at Ocean and Underwater Medical Research and Training Center of ROK Navy, for 6 years Jan. 1993 to Nov. 1998. Result : 1) The mean no-decompression limit excess time between type I DCS group(72.7 min) and type Ⅱ DCS group(92.8 min) showed significant difference. 2) The rate of symptoms appeared on surfacing and within 10min. after surfacing of type Ⅰ and type Ⅱ DCS were 41.4% and 72.7%, respectively 3) The cure rate of type Ⅰ and type Ⅱ DCS were 75.9% and 42.4%, respectively. In type Ⅱ DCS group, the cure rate of the group within 12 hour-delayed recompression treatment and the group above 12 hours-delayed treatment were 64.3% and time 26.3%, respectively, and in type Ⅰ DCS group, 100% and 66.7%, respectively. Conclusion : These findings suggest that the education of safety, the strict observance of the standard decompression table, and the avoidance of excessive repeated diving are important for reducing the risk of diving related disease. And to offer proper management of DCS, there should be more multiplace hyperbaric oxygen chambers, the suitable transport system, and the specialist of diving medicine or hyperbaric medicine in Korea.