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최창운,양승대,우광선,정위섭,임수정,서용섭,전권수,안순혁,이종두,홍성운,임상무 ( Chang Woon Choi,Seung Dae Yang,Kwang Sun Woo,Wee Sup Chung,Soo Jung Lim,Yong Sup Suh,Kwon Soo Chun,Soon Hyuk Ahn,Jong Doo Lee,Sung Soon Hong,Sang Moo Lim 대한핵의학회 1998 핵의학 분자영상 Vol.32 No.3
Purpose : The aim of this sutdy was to evaluate the feasibility of 3-[131I]Iodo-O-methyl-L-a-methyltyrosine ([131I]OMINT) as an agent for tumor image. Materlals and Methods: After synthesis of 4-O-methyl-L-a-methyltyrosine (OMAMT), OMAMT was labeled with 131I using Iodogen method. In viro cellular uptake study was performed using 9 L gliosarcoma cells at various time points upto 4 hr. The biodistribution (five rats implanted with the 9 L gliosarcoma cells per group) was evaluated at 30 min, 2 hr, 24 hr after iv injection of 3.7 MBq [131I]OMIMT or L-3-[131I]iodo-a-methyltyrosine ([131I]IMT). Gamma camera images were obtained at 30min, 2 hr, and 24 hr. Results : [131I]OMINT uptake was 3.3 times and 2.5 times higher than [131I]IMT uptake at 30 min and 60 min, respectively and same after 2 hr in in vitro sutdy using 9L gliosarcoma cells. Maximum accumulation in tumor occurred at 30 min for both [131IOMINT and [131I]IMT in tumor bearing rats. The tumor uptake of [131I]OMINT was significantly higher than that of [131I]IMT in tumor bearing rats. The tumor uptake of [131I]OMIMT was significantly higher than that of [131I]IMT at early time point studied (3.74 +- 0.48 vs 0.38 +- 0.17% ID/g at 30 min and 2.40 +- 0.17 vs 0.24 +- 0.03% ID/g at 2 hr, respectively, p<0.01). However, the tumor uptake of both radiolabels were not significantly different at 24 hr (0.04 +- 0.01 vs 0.05 +- 0.01% ID/g). Tumor was visualized as early as at 30 min in gamma camera images. Conclusion : These data suggested that [131I]OMIMT might be a useful tumor imaging agent and has more advantage for the tumor imaging compared to [131I]IMT. (korean J NuclMed 1998;32;290-7)
송인호 ( In Ho Song ),이태섭 ( Tae Sup Lee ),강주현 ( Joo Hyun Kang ),이용진 ( Yong Jin Lee ),김광일 ( Kwang Il Kim ),안광일 ( Gwang Il An ),정위섭 ( Wee Sup Chung ),천기정 ( Gi Jeong Cheon ),최창운 ( Chang Woon Choi ),임상무 ( Sa 대한핵의학회 2009 핵의학 분자영상 Vol.43 No.5
목적: Hydrodynamic-based procedure는 손쉽고 간편한 비바이러스성 유전자 전달 방법으로 특히 간특이적으로 발현하는 특징을 가진다. 단순 헤르페스 바이러스 제 1 형 티미딘 키나제(herpes simplex virus type 1 thymidine kinase, HSV1-tk)와 다양한 기질을 이용한 비침습적 HSV1-tk 유전자 영상시스템이 널리 연구되어왔다. 본 연구에서는 HSV1-tk 유전자를 hydrodynamic-based procedure를 이용하여 전달한 후, HSV1-tk의 보고 기질로 알려진 5-(2-iodovinyl)-2`-deoxyuridine (IVDU)을 이용하여 간 특이적인 HSV1-tk 유전자 발현 영상을 획득하고자 하였다. 대상 및 방법: HSV1-tk 유전자와 녹색형광유전자를 가진 각 플라스미드 벡터를 마우스에 hydrodynaminc injection을 통해 전달하고, 24 시간 뒤 유전자의 발현을 확인하기 위해 RT-PCR, 생체형광영상, 핵의학영상, 전신자가방사영상 그리고 생체분포를 시행하였다. 결과: 각 플라스미드 벡터를 전달한 간으로부터 추출한 전체 RNA를 이용하여 RT-PCR을 수행한 결과, 각각 HSV1-tk 유전자와 녹색형광단백 유전자의 특이적인 밴드를 관찰할 수 있었다. 생체 분포 결과, pHSV1-tk 벡터를 전달한 마우스의 간에서 특이적인 [123I]IVDU의 섭취를 보였다. 생체형광영상에서는 pEGFP-N1 벡터를 전달한 마우스의 간에서는 유의한 형광신호를 나타내었다. 전신자가방사영상과 감마카메라 영상에서 pHSV1-tk 벡터를 전달한 마우스의 간에서 방사표지 IVDU가 국소적으로 집적되는 것을 확인하였다. 결론: 본 연구에서 hydrodynamic-based procedure는 간특이적으로 플라스미드 DNA를 전달하는데 효과적이며 전달된 유전자의 발현을 분자영상학적인 방법으로 확인하였다. 따라서 Hydrodynamic injection을 통해 HSV1-tk 유전자와 목적 유전자의 공동발현은 방사표지 IVDU에 의해 목적 유전자의 발현을 정량평가하는데 유용할 것으로 기대된다. Purpose: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyl)-2`-deoxyuridine (IVDU) in mouse liver by the hydrodynamic-based procedure. Materials and Methods: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression. Results: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse. Conclusion: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU. (Nucl Med Mol Imaging 2009;43(5):468-477)
L - 3 [ 123I ] iodo - α - methyltyrosine 합성과 9L Glioma 이식 백서 분포조사
임상무(Sang Moo Lim),이종두(Jong Doo Lee),서용섭(Yong Sup Suh),전권수(Kwon Soo Chun),우광선(Kwang Sun Woo),정위섭(Wee Sup Chung),양승대(Seung Dae Yang),임종석(Jong Seok Lim),박현(Hyon Park),윤용기(Yong Ki Yun) 대한핵의학회 1995 핵의학 분자영상 Vol.29 No.1
N/A L-3[123I]iodo-methyltyrosine([123I] IMT) was synthesized by electrophilic radio-iodination using chloramine-T and Iodobead in phosphate buffered solution. And the biodistribution was examined in 9L glioma bearing rats. The radiosynthesis of [123I] IMT with iodobead was simpler and higher in radiochemical yield(88%) than the method using chloramine-T(83%) as radioiodinating reagent. The highest yield was obtained from the reaction using 1 piece of Iodobead, 200μg α- methyltyrosine in 100μl phosphate-buffered solution (pH 5.5) and the reaction was completed in 7min. 24hours after the injection, the biodistribution in 9L glioma transplanted rats revealed the in vivo deiodination, the excretion via kidney, and 3 times higher uptake in the tumor than normal brain. These results suggest the promising clinical use of [123I]IMT in the various ious malignancies.
지방육종형성 동물모델에서 123I-15-(p-iodophenyl)-3-R , S-methylpentadecanoic acid ( BMIPP ) 의 생체분포와 생체영상
최창운(Chang Woon Choi),임상무(Sang Moo Lim),이태섭(Tae Sup Lee),서용섭(Yong Sup Suh),우광선,정위섭(Wee Sup Chung),임수정(Soo Jung Lim),오옥두(Ok Doo Awh) 대한핵의학회 2001 핵의학 분자영상 Vol.35 No.5
N/A Purpose: 123I-labeled fatty acids have been used in the evaluation of regional myocardial energy metabolism. This study aimed to evaluate the usefulness of 123I-BMIPP as a liposarcorna-imaging agent. Materials and Methods: We compared in vitro uptakes between liposarcoma(SW872) and glioma(9L) cell lines, and examined biodistribution and in vivo images of 123I-BMIPP in liposarcoma-bearing nude mice. Cold-BMIPP was labeled with 123I using Cu2+ as catalyst. After purification by Sep-pak, radiochemical purity was determined by TLC. We compared cellular uptake between glioma and liposarcoma after incubation of 5, 10, 15, 30, 60, 120, and 180 mins with culture medium containing I-BMIPP. The difference in biodistribution was determined between non-feeding (water only) group for 18 hr and feeding group in normal mice (n=6/group) at 0.5, 2, and 24 hr. In liposarcoma-bearing nude mice model, liposarcoma, SW872, cell lines were injected subcutaneously into the left thigh of nude mice. The biodistribution of 123I-BMIPP was evaluated at 0.5, 2, and 24 hr (n=5 / group) and in vivo image of 123I-BMIPP was obtained with gamma camera at 2 and 24 hr in liposarcorna-bearing nude mice. Results: Radiolabeling yield and radiochemical purity were 95% and above 99%, respectively. SW872 cell line showed more increased uptake than 9L with 1.5 times at 180 mins. The clearance of 'I-BMIPP in various tissues was more delayed in the non-feeding group than in the feeding group, especially at delayed time (24 hr) in normal mice, and the major excreting organ was the gastrointestinal tract. In liposarcoma-bearing nude mice, tumor/blood ratio of 123I-BMIPP was 0.94, 0.75, and 1.38 and tumor/muscle ratio was 0.66, 1.53, and 1.11 at 0.5, 2, and 24hr, respectively. 123I-BMIPP was selectively localized in liposarcoma at 24 hr image. Conclusions: These results suggest that 123I-BMIPP can be used as a liposarcoma-imaging agent. (Korean J Nucl Med 200135:324-333)
형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가
정재호 ( Jae Ho Jung ),최태현 ( Tae Hyun Choi ),우광선 ( Kwang Sun Woo ),정위섭 ( Wee Sup Chung ),강주현 ( Joo Hyun Kang ),정수영 ( Su Young Jeong ),최창운 ( Chang Woon Choi ),임상무 ( Sang Moo Lim ),천기정 ( Gi Jeong Cheon ) 대한핵의학회 2009 핵의학 분자영상 Vol.43 No.5
목적: 작은 크기의 재조합 단일사슬 항체는 빠른 혈중 제거율과 종양의 항체 집적율이 증가되는 등의 장점을 가지고 있다. 반면에 항체의 작은 크기는 방사성 또는 형광물질의 표지를 위한 킬레이터 결합에 중요한 아미노산 그룹의 감소를 의미하기도 한다. 본 연구에서는 단일사슬 lym-1 염기서열 C-말단에 lysine 아미노산 태그를 삽입하여 형광물질의 직접표지 및 그 표지수율 증가를 확인하고자 하였다. 대상 및 방법: 대장균 pET-22b (+) 벡터에 재조합 된 lysine 삽입 단일사슬 lym-1유전자는 대장균 BL21 (DE3)에 형질전환하여 발현하였다. 생산된 lysine lym-1 항체는 Ni-NTA 컬럽과 분자량 컬럼을 사용해 정제하였고. 단백질 전기영동과 western blot을 통해 확인하였다. lysine lym-1 항체에 방사성 동위원소인 I-124, I-125, I-131 과 Tc-99m를 표지하여 그 수율을 확인하였으며 유세포계측기를 사용해 형광물질인 FITC가 직접표지된 라이신 lym-1 항체의 면역반응성을 사람의 버킷 림프종 세포주인 Raji 세포주에서 면역반응성을 확인하였다. 결과: Lysine 도입 단일사슬 lym-1 항체는 두 과정의 정제를 통하여 획득하였으며 그 크기는 약 48 KDa이었고, 방사성동위원소인 I-124, I-125, I-131과 Tc-99m의 표지수율은 각각 >99%, >99%, >95%, >99%로 확인되었다. 유세포계측을 통한 lysine 도입 단일사슬 lym-1항체의 면역반응성은 기존의 단일사슬 lym-1항체와 유사함을 확인하였다. 결론: 재조합 lym-1 항체에 형광물질을 직접 표지하기 위한 lysine 아미노산의 도입은 항체의 면역반응성 감소를 최소화 시키면서 직접표지 수율을 증가시킬 수 있는 유용한 방법임을 확인하였다. Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FITC conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt`s lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt`s lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity. (Nucl Med Mol Imaging 2009;43(5):487-494)
Alpha - hCG 측정을 위한 섬광 근접 측정법 ( Scintillation Proximity Assay ) 에 관한 연구
최태현(Tae Hyun Choi),최창운(Chang Woon Choi),임상무(Sang Moo Lim),정위섭(Wee Sup Chung),임수정(Soo Jeong Lim),이수진(Su Jin Lee),이태섭(Tae Sup Lee),오옥두(Ok Doo Awh),(Kwang Sun Woo) 대한핵의학회 2002 핵의학 분자영상 Vol.36 No.2
목적: 섬광 근접측정법은 항원 항체 반응 후 결합 분획과 유리분획을 분리하는 광정이 필요없다. 이러한 원리를 검체 내 hCG와 항 α hCG 항체간의 항원 항체 반응에 적용하고자 한다. 대상 및 방법: 항 α hCG 항체를 biotin과 결합시켜 SPA bead에 부착된 streptavidin과 부착 가능하게 만들었다. 이 측정법은 항 α hCG 항체가 부착된 SPA beads에 대해 혈청내 hCG와 표지항원인 [^125I]hCG간의 경쟁 반응을 기본 원리로 이용하였다. Biotin표지 항 α hCG항체를 [^125I]hCG 100㎕와 표준용액이나 환자 혈청 200㎕이 들어있는 실온에서 20분 방치하였다. 그리고 streptavidin이 붙은 SPA beads 20㎕를 바이알에 넣고 10분 더 방치한다. 환자 혈청의 수치를 표준 응답곡선을 통해 계산하였다. 결과: SPA측정법에 사용되는 방사성 핵종의 방사능 양에 따라 반응용액 속에서 SPA bead와 자유 방사성 핵종에 의한 배후 방사능이 측정값에 영향이 없음을 확인하였다. SPA 방법을 응용한 측정에서 적합한 표준 응답곡선을 얻었고, 실제 환자혈청에서의 hCG농도를 결정할 수 있었다. 결론: 이 실험을 통해 SPA 방법을 이용한 측정법이 임상진단에 유용하게 사용될 수 있을 것으로 사료된다. Purpose: Scintillation Proximity Assay (SPA) does not require the physical separation of receptor bound form from free form. SPA was applied to the study of interaction of human chorionic gonadotrpin (hCG) and anti-α hCG in serum. Materials and methods: Anti-α hCG was biotinylated for the binding to streptavidin. The assay was based on the simple competitive binding method between [^125l]hCG and the hCG in sample serum, with anti-α hCG-coated beads. Aliquots of biotinylated anti-α hCG were dispensed into scintillation vials containing 100㎕ [^125ㅣ]hCG and 200㎕ of either a standard concentration of hCG for preparation of standard curve or unknown sample, and oncubated for 20 min. at room temperature. Then 20㎕ streptavidin-coated beads were added to vials. and finaliy incubated for 10 min at room temperature. Values for unkncown samples were then calculated from the standard curve. Results: Optimal backfround counts were certificated using varied radioactivity of radionuclides. Appropriate standard curve was obtained from SPA method successively, and the concentration of hCG from unknown serum was determined by standard curve. The result from SPA assay was similar to that of RIA. Conclusion: This observation confims that SPA method could be useful for cilnical diafnosis.(Korean J Nucl Med 2002;36;133-139)
123I , 99mTc 사람 비특이 IgG 및 67Ga - Citrate의 실험동물에서 염증병소 섭취율의 비교
오옥두(Ok Doo Awh),임상무(Sang Moo Lim),이종두(Jong Doo Lee),우광선(Kwang Sun Woo),정위섭(Wee Sup Chung),서용섭(Yong Sup Seo) 대한핵의학회 1992 핵의학 분자영상 Vol.26 No.1
N/A 123I has ideal half life of 13 hours, suitable 159 keV gamma energy for imaging, and easy labeling methods. In Korea Cancer Center Hospital, 123I has been produced by MC-50 cyclotron. The purpose of this study is looking for good labeling condition of 123I and 99mTc to nonspecific human polyclonal IgG, and comparing these with 67Ga-citrate in the abscess bearing mice. Human polyclonal nonspecific IgG was labeled with 0.2 M phosphate buffer added 123I by chloramine T method. Human polyclonal nonspecific IgG was labeled with 99mTc-gluconate after activation with β-mercaptoethanol. In the abscess bearing mice, the radioactivity in the abscess was higher in 24 hours than 6 hours after injection. In the abscess, 123I nonspecific IgG had higher uptake than 99mTc-IgG or 67Ga-citrate. There was no significant difference in absecess uptake of 123I-IgG among 24, 72, 120 hours abscess age. Further clinical researches with 123I-nonspecific IgG, and other immunoscintigraphies using 123I are expected.