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마우스 동종 조혈모세포 이식모델에서 Cyclosporin A, FK506, 3-Deazaadenosine 등의 약제가 급성 이식편대 숙주병과 생존에 미치는 영향
진종률,정대철,엄현석,정낙균,박수정,최병옥,민우성,김학기,김춘추,한치화,Jin, Jong Youl,Jeong, Dae Chul,Eom, Hyeon Seok,Chung, Nak Gyun,Park, Soo Jeong,Choi, Byung Ock,Min, Woo Sung,Kim, Hack Ki,Kim, Chun Choo,Han, Chi Wha 대한면역학회 2003 Immune Network Vol.3 No.2
Background: We investigated the effect of donor marrow T cell depletion, administration of FK506, cyclosporin A (CSA), and 3-deazaadenosine (DZA) on graft versus host disease (GVHD) after allogeneic murine hematopoietic stem cell transplantation (HSCT). Methods: We used 4 to 6 week old Balb/c ($H-2^d$, recipient), and C3H/He ($H-2^k$, donor) mice. Total body irradiated recipients received $1{\times}10^7$ bone marrow cells (BM) and $0.5{\times}10^7$ splenocytes of donor under FK506 (36 mg/kg/day), CSA (5 mg/kg/day, 20 mg/kg/day), and DZA (45 mg/kg/day), which were injected intraperitoneally from day 1 to day 14 daily and then three times a week for another 2 weeks. To prevent the GVHD, irradiated Balb/c mice were transplanted with $1{\times}10^7$ rotor-off (R/O) cells of donor BM. The severity of GVHD was assessed daily by clinical scoring method. Results: All experimental groups were well grafted after HSCT. Mice in experimental group showed higher GVHD score and more rapid progression of GVHD than the mice with R/O cells (R/O group) (p<0.01). There were relatively low GVHD scores and slow progressions in FK506 and low dose CSAgroups than high dose CSA group (p<0.01). The survival was better in FK506 group than low dose CSA group. All mice treated with CSA died within 12 days after HSCT. The GVHD score in DZA group was low and slow in comparison with control group (p<0.05), but severity and progression were similar with low dose CSA group (p=0.11). All mice without immunosuppressive treatment died within 8 days, but all survived in R/O group (p<0.01). Survival in low dose CSA group was longer than in control group (p<0.05), but in high dose CSA group, survival was similar to control group. The survival benefit in DZA group was similar with low dose CSA group. FK506 group has the best survival benefit than other groups (p<0.01), comparable with R/O group (p=0.18), although probability of survival was 60%. Conclusion: We developed lethal GVHD model after allogeneic murine HSCT. In this model, immunosuppressive agents showed survival benefits in prevention of GVHD. DZA showed similar survival benefits to low dose CSA. We propose that DZA can be used as a new immunosuppressive agent to prevent GVHD after allogeneic HSCT.
혈전성 혈소판 감소성 자반증에서 한국형 혈소판 응집 단백질의 분리 정제 및 특성
진종률(Jong Youl Jin),이종우(Jong Woo Lee),김승호(Seung Ho Kim) 대한내과학회 1995 대한내과학회지 Vol.48 No.1
N/A Objectives: Thrombotic thrombocytopenic purpura (TTP) is characterized by widespread occluding and persistent microthrombotic lesions. Evidence for both endothelial damage and primary platelet aggregation as possible pathogenetic mechanisms has been produced. But the actual mechanism of platelet aggregation has not been explained satisfactorily. Recent studies suggest 37,000 mol wt plateler. aggregating protein (PAP) to be responsible for the formation of platelet thrombi in the patients with TTP. We purified and characterized a novel Korean type 37,000 mol wt platelet aggregating protein from the plasma of a patient with TTP. Methods: K-PAP was purified from the plasma which was obtained during the plasmapheresis in a 31-years- old Korean patient with TTP, by ammonium sulfate fractionation, and DEAE-Sephacel, concanavalin A-Sepharose and Superose 12 gel filtration chromatographies. In each step, platelet aggregating activity was measured by platelet aggregometer. Some biochemical characteristics were studied with PAS stain, sodium dodedyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with or without reduction and inhibition test by normal serum. Results: 1) We found a major protein band with a molecular weight of 37,000 Da and named it as K-PAP. 2) K-PAP seemed a glycoprotein with negative charge, because it bound tightly with DEAF-Sephacel and was stained by PAS. 3) The K-PAP was a single polypeptide because it runned 37,000 mol wt with or without reduction in SDS- PAGE. 4) K-PAP was not tightly bound with concanavalin A. 5) The platelet aggregating activity of K-PAP was inhibited by normal human plasma. Conclusion: In our study, a nouel K-PAP was purifide as a 37,000 mol wt glycoprotein with high platelets aggregating activity from a patient with TTP. It has similar characteistics except the weak binding activity in concanavalin A-Sepharose compared with the purified PAP in foreign reports.
녹색 형광단백 유전자를 함유한 아데노바이러스 주입시 마우스 생체내 발현양상
진종률(Jong Youl Jin),송치원(Chi Won Song),김진아(Jea Na Kim),이희진(Hee Jin Lee),김태규(Tai Gyu Kim),한치화(Chi Wha Han),엄현석(Hyun Seok Eom),박수정(Soo Jeong Park),정대철(Dae Chul Jeong),정낙균(Nak Gyun Chung),김소연(Soh Yeon Kim) 대한내과학회 2001 대한내과학회지 Vol.61 No.5
N/A Background : The green fluorescent protein (GFP) from jelly fish, A equorea victoria, has become a versatile reporter for monitoring gene expression in a variety of cells and organisms . Using GFP as a marker protein we studied whether there are any differencies in the expression patterns among organs in mouse after intravenous injection of adenovirus vectors with GFP gene. Methods : Recombinant E1, E3- defective type 5 adenovirus vectors (2×10(8)/mouse) with CMV promoter and GFP gene were injected into mice via tail vein. On 3, 6, 9, 14, 21, 28 days after gene transfer, 5 mice per experiment s were sacrificed by cervical dislocation and obtained liver , lung, heart , kidney, spleen, small intestine and bone. Half of them were examined by optical microscope after H-E stain. Another half were examined by fluorescent microscope after frozen section. Western blotting were done for each samples with anti- GFP monoclonal antibody and obtained GFP bands were quantitatively compared using Gel-Doc (Bio- Rad, USA) image analyzer. Results : In all organs that we obtained, expression of GFPs are noticed 3 days after gene transfer and reached a maximum around 9th to 14th days, after then the intensities are slightly decreased but maintained until 28th days as determined by Western blotting. On fluorescent microscopic examination, GFPs are well and most frequently expressed on lung among all the examined organs. There are little expression of GFPs on liver parenchymal area around the sinusoids and central veins, although patchy expression of GFPs are observed along the liver capsules. GFPs are highly expressed around the splenic trabecula area but splenic pulp area, it is very spar sely expressed. GFPs are more frequently and highly expressed around the renal tubular area than gromerular area in kidneys. In small intestine, GFPs are expressed on mid portion of microvilli. GFPs are not expressed on myocardium except scanty expression on endocardium. Bone marrow showed GFPs but precise localization is difficult because bony spicules mashed bone marrow during the preparation of frozen section. No specific pathologic lesions possibly related with adenovirus administration are observed on microscopic examination of H-E stained specimens. Conclusions: GFPs can be detected in cells without the fixing and staining and a good marker to studying the kinetics and persistence of adenovirus mediated gene therapy. And there are different GFP expression patterns according to the organs after intravenous injection of adenovirus vectors with GFP gene in mouse.(Korean J Med 61:537- 545, 2001)