비수계 용매에서의 효소반응

지영민 高麗大學校 自然資源科學硏究所 1995 自然資源科學硏究 Vol.3 No.-

효소반응 동력학 정수에 대한 유전상수의 영향

지영민 高麗大學校 自然資源科學硏究所 1998 自然資源科學硏究 Vol.6 No.-

pKT230 벡터를 이용한 Pseudomonas sp. P20 2,3-Dihydroxybiphenyl Dixygenase 유전자의 클로닝

김지영,김치경,가종억,민경희,박용근 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6

Biphenyl과 4-chlorobiphenyl을 분해하는 자연계 분리 균주인 Pseudomonas sp. P20의 chromosomal DNA로부터 pBluescript SK(+)를 이용하여 pcbABCD 유전자를 클로닝하여 재조합플라스미드 pCK1을 제조하였고, 또 pcbCD 유전자를 포함하여 pCK102을 제조하였다. 방향족 탄화수소 화합물의 생분해는 벤젠고리의 개환 과정이 중요하기 때문에, 2,3-dihydroxybiphenyl(2,3-DHBP)의 벤젠고리의 개환에 관여하는 2,3-DHBP dioxygenase(2,3-DHBD) 유전자를 pKT230 벡터를 이용하여 pCK102로부터 클로닝하였다. EcoRI으로 절단한 pCK102와 pKT230 벡터를 ligation시켜 13.8 kb의 hybrid plasmid pKK1을 제조하였다. 2,3-DHBD 유전자를 포함하는 pKK1을 E. coli XL1-Blue에 형질전환시켜 E. coli KK1 재조합 균주를 얻은 후, 2,3-DHBD의 활성을 측정하였다. E. coli KK1의 2,3-DHBD의 효소활성은 pBluescript SK(+)를 이용하여 제조한 재조합 균주인 E. coli CK102의 효소활성과 유사하였으나, Pseudomonas sp. DJ-12와 Pseudomonas sp. P20과 같은 자연계 분리균주보다 훨씬 높았다. Pseudomonas sp. P20 isolated from the polluted environment is capable of degrading biphenyl and 4-chlorobiphenyl. The pcbABCD genes responsible for degradation of biphenyl and 4-chlorobiphenyl were cloned using pBluescript SK(+) from the chromosomal DNA of Pseudomonas sp. P20 to construct pCK1 and pCK102, harbouring pcbABCD and pcbCD, respectively. The 2,3-DHBP dioxygenase gene, pcbC, was cloned again from pCK102 by using pKT230 which is known as a shuttle vector and pKK1 hybrid plasmid was constructed. The E. coli KK1 transformant obtained by transforming the pKK1 into E. coli XL1-Blue showed 2,3-DHBP dioxygenase activity. The specific 2,3-DHBP dioxygenase activity of E. coli KK1 was similar to that of the E. coli CK102, but much higher than those of the natural isolates, Pseudomonas sp. DJ-12 and Pseudomonas sp. P20

부신 결핵에 의한 Addison씨병 1예

송치운,구본정,안봉수,전준식,안미애,이진홍,송민호,김영건,노흥규 충남대학교 의과대학 지역사회의학연구소 1995 충남의대잡지 Vol.22 No.2

Addison's disease is a rare primary adrenal insufficient disorder resulting from chronic deficiency of adrenal cortical hormones. Clinical manefestations are generalized weakenss, weight loss, hyperpigmentation (especially sun exposed area and mucous membrane), hypotension, hyponatremia, hyperkalemia, gastrointestinal symptoms (involving anorexia, nausea, vomiting, abdominal pain). A 34-year-old woman has experienced slowly progressive generalized weakenss and skin pigmentation, anorexia, nausea, vomiting with ascites and diffuse abdominal pain. On the time of admission, her main clinical manifestations were anorexia, nausea, vomiting, fatigue, generalized weakness, amenorrhea, hair loss, diffuse abdominal pain revealed as Addison's disease due to bilateral adrenal tuberculosis. Her adrenal insufficient symptoms were recovered with the replacement of adrenocortical hormones and antituberculous medications. After treatment, Her skin pigmentation was decreased and menstruation was reappeared. Here we experienced one cases of Addison's disease with tuberculous peritonitis.

Mo¨ssbauer분광법에 의한 Ca_(0.75)Ba_(0.25)Fe_2O_4의 Fe이온에 대한 연구

홍치유,윤인섭,조수열,김영민,황진철 東國大學校 1992 東國論叢 Vol.31 No.-

The Mo^ssbauer spectrum of CaFe_2O_4, consists of paramagnetic two doublets absorption peaks at room temperature. But, the replacement of Ca^+2 ions in CaFe_2O_4 with Ba^+2 ions changes the magnetic properties without changing the crystal structure and the Mo^ssbauer spectrum of Ca_0.75Ba_0.25Fe_2O_4 consists of paramagnetic two doublets and ferrimanetic three different Zeeman sextets absorption peaks due to the Fe^57 nuclei in the various sublattices. According to this study, the Fe ions in the Ca_0.75Ba_0.25Fe_2O_4 are trivalent in al sublattices and there are more than three sublattices in this structure.

혈액투석중인 만성신부전 환자에서 골대사 지표로써의 Osteocalcin치

송치운,이진홍,안미애,윤환중,윤상임,성기양,이강현,송민호,이강욱,신영태,김영건,노흥규 충남대학교 의과대학 지역사회의학연구소 1993 충남의대잡지 Vol.20 No.2

Background : Serum osteocalcin is synthesized by osteoblast and has been shown to be sensitive indicator of bone turnover inpatients with various metabolic bone disease. In renal osteodystrophy, serum osteocalcin is elevated due to decreased renal clearance and elevated level of PTH. This study was done to evaluate the usefulness of serum osteocalcin as a marker of bone metabolism and the correlation with other biochemical markers of bone metabolism. Methods : We measured serum osteocalcin, calcium, phosphorus, ALP(alkaline phosphatase) and PTH(parathyroid hormone) in 37 patients with end stage renal disease on hemodialysis. Osteocalcin was determined by radioimmunoassay and PTH was determined by radioimmunometric assay. Results : 1) The mean level of serum osteocalcin in ESRD patients was 233.8± 218.2ng/ml which was significantly higher than that of controls(p<0.0001). 2) The mean level of serum PTH in ESRD patients was 40.5± 43.8pg/ml was significantly higher than that of controls(p<0.005). 3) There was a significant positive correlation between the level of serum PTH, ALP and the level of serum osteocalcin in ESRD patients. 4) By using multiple regression, PTH is most reliable factor that affect to elevated level of serum osteocalcin ( beta coefficient = 0.687, Sig T<0.05). Conclusion : Serum osteocalcin as a marker of bone metabolism in ESRD patients is more useful than other biochemical marker such as serum calcium, phosphorus, ALP and PTH is a most reliable factor that affect to elevated level of serum osteocalin.

산채류로부터 혈소판응집 억제물질의 검색

윤민호,임치환,오진환,이종철,최우영 충남대학교 농업과학연구소 1997 농업과학연구 Vol.24 No.2

To select potential inhibitors of platelet aggregation form large numbers of crude plant extracts, the modified thin smear method for the anti-platelet aggregating activity using platelet rich plasma was further modified by direct observation under a light microscope without staining the smear. The activities determined by the method were coincided with those by the electrical impedence method using whole blood, when ADP or collagen was employed as the aggregating agent. Among 130 varieties of edible and herbal plants which collected from markets or experimental farms of agricultural research institutes, those showed the anti-platelet aggregating activities were selected by testing the crude methanol extracts: Aster scaber, Aster tataricus, Ligularia stenocephala, Platycodon glaucum Allium victorials, Allium oderum, Morus bombycis, Portulaca oleracea, Acanthopanax sessiliflorus and Rosa davurica. However, some of them activated the platelet aggregation under the same assay conditions: Pimpinella brachycarpa, Hosta plantaginea. Capsella bursapastoris, Fagopym esculentum, Prunus mume, Rubus coreanus and Limaria japonica. In addition, those revealed the antioxidant activities were selected by measuring the abilities to scavenge superoxide anion radicals: Pteridum aquilinum, Aster scaber, Ligularia fischeri, Chrysanthemum zawadskii, Artemisia capipparis, Cirsium setidens, Commelina communis and Capsella bursapastoris among edible plants.

당뇨병에 합병한 고지혈증에 대한 Acipimox(Olbetam)의 효과

김영건,김진희,전준식,송치운,송민호 충남대학교 의과대학 지역사회의학연구소 1993 충남의대잡지 Vol.20 No.2

Multiple abnormalities of lipoprotein metabolism exist in patients with diabetes mellitus that could contribute to their enhanced susceptibility to atherosclerosis. Acipimox(5-methyl-pyrazine carboxylic acid 4-oxide, Farmitala Carlo Erba, Milano Italy) is a new potent and long acting(8h) antilipolytic agent, which has been derived from nicotinic acid. By lowering plasma and possibly skeletal muscle concentrations of non-esterified fatty acid, Acipimox may have a potential beneficial effect on glucose metabolism in patients with non-insulin dependent diabetes mellitus. The aim of this study was to evaluate the acute effect of Acipimox on hyperlipidemia associated with non-insulin dependent diabetes mellitus. 18 patients with non-insulin dependent diabetes mellitus were in the study. They had fasting plasma triglyceride concentrations above 250 mg/dl and cholesterol concentrations above 240 mg/dl. The level of triglyceride, cholesterol and glycated hemoglobin were significantly reduced after 8 weeks treatment of Acipimox. Adverse effects were noted in 3 patients, but it was transient with the course of administration. Acipimox is an effective and safe drug for the management of hyperlipidemia associated with non-insulin dependent diabetes mellitus.

백혈병 마우스 모델의 동종골수이식에서 활성화된 자연살해세포들의 보충이 이식편대백혈병효과와 이식편대숙주반응에 미치는 영향

엄현석,한치화,박수정,김소연,정낙균,정대철,진종률,최일봉,양형모,서영훈,송현근,최인표,민우성,김춘추 대한조혈모세포이식학회 2001 대한조혈모세포이식학회지 Vol.6 No.1

배경: 백혈병에서 동종골수이식 (allogeneic bone marrow transplantation)의 성공적 치료 효과를 얻기 위해서는 이식편대숙주반응 (graft-versus-host disease, GVHD) 발생의 극복과 재발의 방지가 중요한 과제이다. 골수를 역류원심성 세포분리 (counterflow centrifugal elutriation, CCE) 방법으로 분리하여 얻은 rotor off (R/O) 세포분획은 T 세포의 수는 적지만 조혈모세포들을 다량 포함하고 있어 동종골수이식에서 주조직적합복합체 (major histocompatibility complex, MHC) 차이를 극복할 수 있고, 이식편의 생착 성공과 GVHD 발생 예방에 효과적이다. 그러나 골수로부터 T 세포를 제거하면 백혈병세포를 공격하는 이식편대백혈병 (graft-versus-leukemia, GVL) 효과가 감소되기 때문에 백혈병 재발의 빈도가 높다. 자연살해세포 (natural killer cell, NK cell)의 보충 첨가는 동종골수이식 후 GVHD 발생을 줄이면서 충분한 GVL 효과를 얻을 수가 있다. 따라서 저자는 분리 후 IL-2로 활성화시킨 NK 세포들을 골수 R/O 세포분획과 함께 백혈병 마우스 모델에 동종이식함으로써 GVHD와 GVL에 미치는 효과를 관찰하였다. 방법: Balb/c (H-2^(d)) 마우스에서 유래된 A20 (murine B-lymphoma/leukemia cell line, H-2^(d)) 백혈병 세포를 이식 2 일 전에 Balb/c 마우스에 주입하고, 치사량의 전신 방사선을 조사한 직후에 Balb/c 또는 C57BL/6 (H-2^(b)) 마우스의 골수 R/O 세포분획을 꼬리정맥을 통하여 주입하였다. 이들은 모두 이식 후 6-8 주 이내에 사망하였다. 동종이식의 대조군 (n=9)에는 1 × 10^(7)의 R/O 세포분획만을 주입하였고, 실험군 (n=9)에는 C57BL/6 마우스의 비장세포들로부터 단클론항체들을 이용한 negative selection방법으로 분리한 후 IL-2로 활성화된 5 × 10^(5)의 NK 세포분획을 1 × 10^(7)의 R/O 세포분획과 함께 주입하였다. GVL 효과의 판정은 이식 후 14 일과 28 일 째 되는 날 마우스에서 골수, 비장, 간 등을 얻어 백혈병 세포들의 침윤을 조직학적으로 관찰하였으며, GVHD의 정도는 육안적 관찰법으로 평가하였다. 결과: R/O 세포분획만을 이식한 대조군의 골수, 비장, 그리고 간 조직에서는 A20 백혈병 세포의 침윤이 각각 89% (8/9), 78% (7/9)와 22% (2/9)에서 관찰되었고, R/O 세포분획과 NK 세포분획을 함께 이식한 실험군에서는 비장과 간을 제외한 골수에서만 89% (8/9)에서 A20 백혈병 세포의 침윤이 관찰되어 두 군 사이에 장기별 분포의 차이를 볼 수 있었다 (P= 0.0001). 한편 GVHD는 두 군 모두에서 경하게 나타나서 유의한 차이는 없었다. 또한 생착 부전으로 사망한 마우스는 없었다. 결론: CCE를 이용하여 T 세포를 제거한 동종골수이식에서 NK 세포의 보충은 GVHD의 악화는 일으키지 않으면서, 백혈병의 진행을 억제하는 GVL 효과를 얻을 수 있었다. Background: Allogeneic bone marrow transplantation (BMT) with T cell-depleted marrow accompanies engraftment failure and relapse of leukemia by a loss of the graft-versus-leukemia (GVL) effect frequently, while it can prevent GVHD. Supplement of NK cells could prevent GVHD and enhance GVL effect in several murine allogeneis BMT models Roter off (R/O) cell fraction obtained by counterflow centriation elutriatio (CCO) contains small number of T cells and many hematopoietic stem cells. The aim of this study was to determine the effect of R/O cell fraction supplemented with IL-2 activated NK cells on GVL and GVHD within the leukemic mouse BMT model. Methods: Inoculation of A20 (H-2d, murine B-lymphoma/leukemia, Balb/c origin) cells into Balb/c mice via the tail vein 2 days prior to lethal total body irradiation (TBI) and infusion of the Balb/c BM or C57BL/6 (H-2b) R/O fraction were performed. It resulted in 100% mortality within 6 to 8 weeks. The irradiated mice in the control group were injected with 1 × 107 R/O cell fraction alone (n=9) and in the experimental group mice were injected with 1 × 107 R/O cell fraction plus 5 × 105 negatively selected IL-2 activated NK cell fractions of the spleens via the tail vein (n=9). On day 14 and 28 after BMT, the bone marrows, spleens, and livers of mice were harvested for histopathologic analysis of the infiltrations of leukemic cells. We then evaluated the GVHD within the mice. Results: A histopathologic study of the recipients receiving R/O fraction alone showed infiltration of leukemic cells, 89% (8/9) in bone marrows, 78% (7/8) in spleens, and 22% (2/9) in livers. The experimental group of mice showed only the infiltration of leukemic cells 89% (8/9) in bone marrows, not in spleens and livers. There were the organ differences of the leukemic cells infiltrations between the two groups (P=0.0001). There were no obvious differences in the GVHD scores between these two groups, and severe GVHD was not observed. There was no engraftment failure among groups. Conclusion: Thus, our findings suggest that R/O cell fraction obtained by CCE and supplemented with NK cells can promote GVL effect without mediating clinically overt GVHD in allogeneic BMT of mouse leukemia.

관상동맥내 스텐트 삽입후 발생한 관상동맥류 2례

제인수,허민영,문치숙,이재호,정수룡,조길현,이영민,김대경,김두일,김동수 인제대학교 백병원 2002 仁濟醫學 Vol.23 No.3

Coronary artery aneurysms are detected with increasing frequency because of the routine use of coronary angiography in the diagnosis and management of coronary artery disease. Although some coronary artery aneurysms are congenital or inflammatory, most are atherosclerotic. The prognosis of atherosclerotic coronary artery aneurysm may depend on the morphology of the aneurysm or the presence of coexisting obstructive disease. Coronary artery aneurysms have been also reported to occur after ballon angioplasty, directional athrectomy, laser angioplasty, and stent implantation with a frequency of 2% to 4%. The long-term outcome of post-intervention coronary artery aneurysm is unknown. We report two cases of the development of a coronary aneurysm 6 months after stent implantation as a treatment of severe obsructive coronary artery disease.