인삼엽에서 superoxide dismutase Activity 측정 및 Isozyme pattern 검정

양덕조, 김명식, 이성종 忠北大學校部設基礎科學硏究所 1987 自然科學硏究 Vol.1 No.1

-본문참고-

Alkaline Phosphatase Isoenzyme의 Cellulose Acetate Microzone 電氣泳動 및 組織化學的 檢索

楊聖基,金在植,金重明 慶北大學校 醫科大學 1979 慶北醫大誌 Vol.20 No.2

正常 成人의 血淸 ALP 活性은 主 分劃이 α_2 肝分劃이었고, 副 分劃은 α_2 /β骨分劃을 나타내었고 發育期의 小兒에서는 骨分劃이 主 分劃으로 나타났고, 肝分劃은 副 分劃이었으며, 新生兒 臍帶血淸에서는 骨分劃만을 主로 나타내었으며, 胎兒에서는 骨分劃과 가장 移動度가 바른 Alb分劃을 나타내었고, 姙婦의 血淸에서는 骨分劃과 같은 移動度를 보였으나 熱에 安定한 胎盤分劃을 나타내었다. 各種 病患別로 ALP CAM 血淸電氣泳動像에서 肝癌은 主로 肝分劃, 骨分劃 및 α_1分劃을 보였고 急性바이러스性肝炎은 主로 肝分劃, 骨分劃 및 Alb分劃을 나타내었고, Choloangioma는 主로 肝分劃과 骨分劃에 α_1과 Alb分劃을 同時에 나타내어 比較的을 特異하였고, 骨病患에서는 거의 特徵的으로 骨分劃을 나타내었고, 胃腸病患과 肺硬化症에서 드물게 特異的 腸分劃을 타타내었다. L+α_1分劃像을 나타낸 肝癌 69例中 原發性肝癌의 從來의 診斷手段을 통하여 cold area만 시행되었던 例가 35例(50.7%), 符合되었던 例中 cold area에 肝針生檢 또는 肝針吸引細胞診 供行例가 15例(21.7%) 細胞診만 施行된 例가 2例(2.9%)이었으며, 이것은 肝癌에 特異하다고 보았다. 各種 臟器組織의 抽出液에 對한 ALP CAM 電氣泳動像에서 肝臟은 血淸의 肝分劃에 該當되는 肝分劃을, 骨은 骨分劃을, 胎盤은 胎盤分劃을, 腸은 腸分劃을, 그리고 腎臟은 腎分劃을 보여 주었고 腎分劃은 血淸에서 아주 드물게 나타나는 것 같았다. 名種 臟器組織에 있어서 ALPI의 活性度는 凍結簿切標本의 肉眠的 所見으로 胎盤, 肝, 腎, 腸, 脾의 順으로 낮았고, 肉筋은 陰性이었으며, 腎臟은 皮質에만 陽性이었고, 鏡檢으로 肝은 主로 bile canaliculi에 極度의 活性을 보였고 肝細胞內에도 若干 認定되었고, 腎臟은 皮質의 近位曲細尿管上皮細胞屬에 ALP活性을 極度로 나타내었으며, 胎盤은 syncytial troplast에 ALP活性을 極度로 나타내었고, 脾臟은 red pulp의 sinusoid에, 그리고 腸은 粘膜屬에만 中等度로 活性을 나타내었고, 空腸에서는 極度로 ALP活性을 보였다. 白血球增加症 例의 미소백혈구 에는 細胞質內에 多樣하게 顆粒狀의 ALP活性을 보였다. 姙娠週數에 胎盤組織과 姙婦血淸에 對한 ALP CAM 電氣泳動 및 組織化學的 檢索에서 ALP 活性은 共히 姙娠週數에 比例하여 增加하였고 末期에 極度에 達하였다. The cellulose acetate microzone electrophoreses of CAME (ALPI) were performed on the sera and various organ tissues, to estimate the source of ALP in the serum of normal physiological and pathological conditions. On the ALP CAME finding, normal adult serum revealed α_2 fraction(liver)as the major fraction and α_2/βfaction (bone) as minor, However, opposite results were obtained from sera of growing child and newborn infant. Sera of premature infants revealed the fastest moving albumin fraction (Alb) and α_2βfraction, and sera of pregnant women, especially in the third trimester of pregnancy revealed heat stable α_2βfraction (placenta) as the major ones and trace of α_2 fraction as the minor. On the ALPI CAME finding in various diseases, liver and α_1 pattern(L+α_1) mainly confined to hepatoma in 94.0 percent, followed by liver abscess in 88.9%, liver and Alb pattern (L+Alb )to acute viral hepatitis and extrahepatic obstructive jaundice in 100% respectively, liver, Alb and α_1 pattern (L+Alb+α_1)to cholangioma in 87.5% liver and β/γ(intestine) pattern(L+I) only to liver cirrhosis, and liver and bone pattern (L+B) to bone diseases only. Therefore these patterns seemed to be specific in the clinical diagnosis. On the ALP CAME findings of various organ-tissue extracts, liver revealed liver fraction, bone and spleen bone fraction, kidney γ/ fraction (kidney) as well as liver fraction as a minor fraction, placenta heat stable α_2/β(placenta) as a major fraction, intestine of fraction, the lung broad fraction from liver to intestineal fraction. However hepatoma and brain tissues did not showed any fraction at all. On the gross inspection of stained slides of ALP from various organ-tissues, such as placenta of the third thrimester pregnancy, kidney, liver, spleen and lymph node, revealed diffuse and strong activity, lung and thyroid revealed diffuse and moderately strong activities, the mucosa of jejunum and the lobule of the breast, revealed strong activities, the mucosa of stomach, colon, rectum, gall bladder and endometrium moderate activities, muscle did not showed any visible activity. Microscopically the organ-tissue ALP were observed mainly in bile canaliculi and portal triad, syncycial bud and sprout of placenta, mucosal cell of jejunum and around the mucosal cells of stomach, colon, rectum, proximal convoluted tubule of renal cortex, osteoblast, sinusoid of splenic red pulp, around the germinalcenter in lymph node, intima of blood vessel, alreolar cell in lung, colloidal follicle of thyroid gland, and endometriu in. Along with progression of gestational weeks, ALP activities of both serum and placental tissue were increased gradually and reached maximum at 40 weeks gestation. The source of elevation of serum ALP activity seemed to be originated mainly from diseased liver and bone, infrequently from kidney and intestine, and possibly from the other tissues.

카라기난 분해효소 생산균의 분리, 동정, 및 효소생산 최적 조건

양승택,주동식,박중제,이정석,김명식,이응호 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6

해양 동식물, 토양 등으로부터 분리한 카라기난 분해능이 확인되었던 80여 균중에서 환원당 생성능이 가장 높았던 strain no.43 균주를 최종 시험균주로 선택하여 이 균주의 최적 효소 생산 조건을 실험하였다. 질소원으로는 nutrient broth 0.7% 농도가 가장 적절한 조건이었고, 탄소원의 농도는 카라기난 0.2%가 가장 높은 효소활성을 나타내었다. NaCl 농도는 1.5%, pH, 온도는 각각 7.0, 30℃로 이 조건에서 96시간 배양하는 것이 가장 적절한 효소 생산 조건이었음이 확인되었다. 각종 생화학 실험을 통해 동정을 행한 결과 그람음성 간균으로 oxidase, catalase 생성의 호기성균, indole 비생성, proline, arginine, serine, citrate, lactate 이용성, gelatin 액화, glucose, fructose, maltose 비이용, mannitol 이용 등의 특성으로부터 Pseudomonas alcaligenes로 동정할 수 있었고 Pseudomonas alcaligenes JCL-43으로 명명하였다. The 80 strains which produce carrageenan degrading enzyme were isolated from soils, mud, seaweed, marine moluscus and echonodermata samples. Among them, one isolated strain, which showed the highest activity to produced carrageenan degrading enzyme, was used for this study. The isolated strain was identified as Pseudomonas alcaligenes through its morphological, biochemical, and physiological characteristics. The best conditions for enzyme production were 0.7% nutrient broth and 0.2% carrageenan as nitrogen and carbon source, respectively. The optimal pH, NaCl, temperature and culture time for carrageenan degrading enzyme were 7.0, 1.5%, 30* and 96hrs, respectively.

Craves병 환자에서 백혈구와 갑상선 조직의 HLA-DR β 유전자 부위의 비교

양인명,우정택,팽정령,서광식,김성운,김영설,김광원,최영길 경희대학교 유전공학연구소 1989 遺傳工學論文集 Vol.1 No.-

정상 갑상선세포에서는 HLA-DR 항원이 발현되지 않았으나, 그레이브스병 환자의 갑상선세포 표면에는 HLA-DR 항원이 발현됨이 보고되어, 이러한 현상은 이 질환의 자가면역 기전에 중요한 역할을 하고 있을 것으로 생각되고 있다. 한편 최근 DR이나 DQ 유전자의 상부 159kb 이내에는 이들의 발현을 조절하는 유전자가 존재함이 알려져 있고, 이 부위의 구조적인 변화로 인하여 DR DQ 유전자의 발현을 유도하는 여러 가지 핵내 인자들과 interferon-r 와 같은 외부인 자들이 보고되고 있다. 그러나 그레이브스병에서 이들 유전자 부위의 구조적인 변화에 관해서는 아직 보고가 없다. 이에 본 연구자 등은 이러한 가능성 여부를 규명하고자 2명의 전형적인 그레이브스병 환자의 수술 우 얻어낸 갑상선 조직에서 RNA를 분리한 우 DR β유전자를 소식자로 northem blotting을 하여 mRNA의 발현을 관찰하였으며, 말초혈액 백혈구와 갑상선 조직으로부터 분리된 DNA를 EcoRI BamHI. HindⅢ PvuⅡ TaqI, PstI등의 6가지 제한효소로 소화한 후 DR β유전자를 소식자로 하여 RFLP 양상을 비교한 결과, 환자 모두에서 mRNA의 발현이 관찰되었으나, 환자 모두에서 6가지 제한효소에 의한 RFLP 양상이 동일하였다. 이러한 결과는 Graves병 환자의 DR 유전자의 발현에 있어서 이 유전자 부위의 구조적인 변화가 관여할 가능성이 적음을 시사하는 사실이라고 사료되나 향후 더 많은 예와 더 많은 제한효소를 이용한 주시가 필요할 것이다. The requirement for major histocompatibility antigen class Ⅱ molecules in the recognition of antigen by helper T cells suggests that the expression of class Ⅱ antigen may be important in the initiation and prolongation of immunopathology. HLA class Ⅱ antigenes are expressed on the surface of thyrocytes of the patients with graves disease. The increased expression of class Ⅱgene can be induced by trans acting factor such as interferon However the possibility of rearrangement of their regulatory genes has not been explored so far. We studied the mRNA expression in the thyrocytes of 2 patients with Graves' disease and compared the restriction fragment length polymorphism (RFLP) between thyroid and peripheral leukocyte DNA. The prominent expression of mRNA was observed in the thyroid tissues of all the two patients. But we did not find any difference in RFLP pattern in both patients. These results suggest the possibility that the rearrangement of the regulatory gene located in the upstream of DR- β gene can be a role in expression of DR antigen is less likely.

흰쥐 샘뇌하수체의 면역전자현미경적 연구 : 젖샘자극호르몬 분비세포와 성장자극호르몬 분비세포

하상선,박경호,양남길,안의태,고정식 순천향의학연구소 1995 Journal of Soonchunhyang Medical Science Vol.1 No.2

본 실험은 흰쥐 샘뇌하수체의 세포형태를 밝히기 위하여 이중면역금입자표지법을 이용하였다. 체중 200-250g의 Sprague Dewley계 숫흰쥐를 실험동물로 사용하였다. ether로 마취된 후 뇌하수체전엽을 적출하여 1% glutaraldehyde- 1% paraformaldehyde액으로 일차고정하였으며, 2% osmium tetroxide액에 이차고정하였다. 고정이 끝난 조직은 alcohol과 propylene oxide로 탈수한 후 araldite혼합액에 포매하였다. 포매된 조직은 LKB-V ultratome으로 60-70cm두께의 얇은 절편을 작성하여 300 mesh nickel grid에 붙인 다음 면역염색 및 이중면역금입자표지법을 시행하였다. 젖샘자극호르몬에 대한 면역염색은 sodium m-periodate로 45분간 처리한 다음, 비특이적 면역반응을 제거하기 위해서 bovine serum albumin(BSA, Sigma) 1% 용액을 사용하였으며, 완충용액으로는 Tris buffered saline, pH 8.2(TBS; 20mM Tris buffered + 20mM NaCl + 0.01% sodium azide)을 사용하였다. 일차항체는 rabbit anti ovine prolactin(ICN Chemicals)을 1 : 3,000으로 희석하여 사용하였으며, 이차항체는 biotin이 표시된 goat anti rabbit Ig G(희석비율 1 : 500, Amersham)을 사용하였고, 금입자표지는 streptavidin gold(희석비율 1 : 100, 10nm, Amersham)을 사용하였다. 성장자극호르몬에 대한 면역염색은 sodium m-periodate로 45분간 처리한 다음, 완충용액으로는 phosphate buffered saline, pH 7.4(0.01M)을 사용하였다. 비특이적 반응을 줄이기 위하여 100mM ammonium chloride로 처리한 후 rabbit anti human growth hormone(BioGenex)을 1 : 40으로 희석하여 면역 반응을 시킨후, protein A-gold(희석비율 1 : 50, 5 nm, BioCell)로 금입자표지를 하였다. 이중면역염색은 Bendayan(1982)의 방법을 변형하여 시행하였는데, grid의 한쪽 면은 젖샘분비자극호르몬의 항체, 다른 쪽 면은 성장자극호르몬의 항체로 반응시켰다. 면역염색이 끝난 절편은 uranyl acetate와 lead citrate로 염색한 후 JEM 100 CX-Ⅱ 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 흰쥐 샘뇌하수체를 이중면역금입자표지법을 이용하여 젖샘자극호르몬분비세포는 3종류, 성장자극호르몬세포는 2종류로 구분할 수 있었다. 1. 젖샘자극호르몬분비세포는 불규칙한 모양을 한 큰 분비과립(300-700 nm)을 가진 성숙세포, 크기가 다양한 둥근 분비과립(150-200 nm)을 가진 중간 세포와 크기가 작은 둥근 분비과립(100-150 nm)을 가진 미성숙세포로 나눌 수 있었다. 2. 성장자극호르몬분비세포는 크고 둥근 분비과립(200-500 nm)을 가진 제 1 형 세포와 상대적으로 작고 둥근 분비과립 (150-200 nm)을 가진 제 Ⅱ형 세포로 나눌 수 있었다. 3. 젖샘자극호르몬과 성장자극호르몬이 한 세포내에 함께 존재하는 경우는 관찰 할 수 없었다. This experiment was aimed at clarifying immunocytochemical characteristics of growth hormone cells and prolactin cells in male rat adenohypophysis, using double immunogold method. Under ether anestehesia, male rats weighing 200-250 gm were decapitated. Adenohypophysis were fixed in the 1% glutaraldehyde - 1% paraformaldehyde solution. followed by refixation in the 2%osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture. The sections were cut on a LKB V ultrotome, and ultrathin sections were placed on nickel rid(300 mesh). The section-bearing grids were floated upside down on th esolutions in a moisture chamber at room temperature. Sections were etched with a saturated solution of sodium m-periodate for 45 min. Aftr etching. sections were pretreated with 0.02M Tris buffered saline(TBS), pH 8.4, with 1% bovine serum albumin(BSA, Sigma) for 60 min, then treated with rabbit anti-sheep prolactin(ICN incubated 60 min in biotinylated goat anti-rabbit IgG(Amersham) diluted 1:100 in TBS with 0.1% BSA. Then sections were incubated on streptavidin gold rinsed with TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the streptavidin gold step, the sections were jet washed with distilled water. According to Bendayan(1982) method, the opposite side of the grid was etched with saturated solutin of sodium m-periodate for 45 min. Aftr etching, sections were treated with 0.01M phosphate buffered saline(PBS), pH 7.4, with 0.1% BSA and 0.1M ammonium chloride for 60 min, then treated with rabbit anti-human growth hormone(BioGenex) diluted 1:40 in PBS with 0.1% BSA for overnight. Grids were incubated 60 min in protein A-gold(5 nm, BioCell) diluted 1:50 in PBS with 0.1% BSA. The sections were jet washed with distilled water. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100CX II electron microscope. The results were as follow: Three types of prolactin cells and two types of growth hormone cells of the rat adenohypophysis were recognized by double immunogold electron microscopy. 1. Matrue prolactin cells are characterized by irregulary shaped large secretory granules (300-700 nm): intermediate type cells contain round granules of varying sizes(150-200 nm): and immature type cells have small round secretory granules(100-150 nm). 2. Type I growth hormone cells are characterized by large round secretory granules(200-500 nm): type II growth hormone cells are characterized by large round secretory granules(150-200 nm). 3. In the male rat adenohypophysis double immunogold labeled with 10 nm gold particles for prolactin and with 5 nm gold particles for growth hormone proved that growth hormone and prolactin were not contained in the same cell at the same time.

어린이에서 안검봉합시 흡수봉합사의 이용

김상덕,양연식,김재덕 圓光大學校 醫科學硏究所 1995 圓光醫科學 Vol.11 No.1

This prospective clinical study was conducted to observe the outcome of surgical skin wound closure in the lower eyelid using absorbable suture material in young children. We operated 30 eyes of 15 patients for entropion and epiblepharon using 6-0 chromic catgut. The patients were carefully examed 1 day, 7 days, 10 days, and 3 months postoperatively for inflammation, wound infection and grossly visible scar. During the follow-up, 6-0 chromic catgut was absorbed and spontaneously disappeared between 7 days and 10 days. There were no inflammation and wound infection in all cases. But the partial visible scar by wound dehiscences was noted in one case. Considering these finding, we think that skin closure with chromic catgut has the advantages of the avoidance of patient discomfort, suture removal, and time spared for the surgeon. Especially, this is extremely helpful in young children.

廢 乾電池로 부터 有價金屬의 循環利用

李成植,金種和,姜奉根,梁種奎 동아대학교 환경문제연구소 1991 硏究報告 Vol.14 No.1

The recovery and recycling of the rare metal from spent batteries have been investigated. It was proposed the synthetic processes of recovery of rare metal from various spent batteries, and spent batteries have to be considered as a resource and the collection system with separation should be established.

글루카민형 킬레이트 수지에 의한 붕소의 분리·농축에 관한 연구

이성식,양종규,김종화 동아대학교 공과대학 부설 한국자원개발연구소 1994 硏究報告 Vol.18 No.1

A process was studied for separation and concentration of boron from steelmaking dust by the selective leaching with sodium hydroxide and adsorption and desorption with chelating resin. The major constituents in this steelmaking dust are iron, zinc, sodium and silicon. The steelmaking dusts were contented of rare metals such as boron, gallium and vanadium. Of the boron contained in the dust, 72% leached with 3.0㏖/1 sodium hydroxide solution. To remove the impurity metals, they were precipitated as their hydroxides by adjusting the solution pH being 10. The selective concentration of boron was done using a chelating resin column with a functional group of glutamine type, and the eluate containing 4.7g/1 of boron was obtained.

봉독약침이 류마티스 관절염 환자의 기능회복 및 삶의 질에 미치는 영향

이상훈,이현종,박상민,김수영,박재경,홍승재,양형인,이재동,최도영,김건식,이두익,이윤호 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2003 東西醫學硏究所 論文集 Vol.2003 No.-

Objective: To evaluate the effects of bee venom acupuncture(BVA) on the rehabilitation and quality of life in rheumatoid arthritis(RA) patients Methods: Patients with RA were treated with the BVA therapy twice a week for 3 months. Tender joint counts, swollen joint counts, morning stiffness, Erythrocyte Sedimentation Rate(ESR), C-reactive protein(CRP), patient global assessment, physician global assessment, Korean health assessment questionnaire(KHAQ) were estimated and analyzed before and after BVA therapy. Results: Tender joint counts, swollen joint counts, morning stiffness showed significant decrease after BVA therapy. But, as acute inflammatory reactants, ESR showed no significant difference and CRP showed significant increase after BVA therapy. Patient global assessment physician global assessment, and KHAQ index showed significant improvement after BVA therapy. Conclusions: BVA therapy can improve rehabilitation and health-related quality of life RA patients as well as clinical symptom and signs. Further study is required in more population with large scale including acute inflammatory reaction of BVA therapy.

신경병증성 통증 모델쥐 뒤뿌리신경절의 Nitric Oxide Synthase 아형 및 NADPH-diaphorase 발현에 관한 연구

정성균,김수일,양은진,류택근,권오유,박경란,이영호,김원식 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.2

Injury to peripheral nerve somtimes lead to neuropathic pain. Nitric oxide(NO) is increased in the dorsal root ganglia (DRG) of neuropathic pain models, and L-NAME, a nitric oxide synthase (NOS) inhibitor, reduces neuropathic pain. However, it is not known that underlying mechanism of NO involvement in development and maintanence of neuropathic pain. It was found that there were three NOS isoforms-nNOS, eNOS, and iNOS. It is also necessary to investigate NOS isoforms in neuropathic pain. The aim of this study is to investigate NOS isoforms and NADPH-diaphorase expressions and underlying mechanism of NO involvement in neuropathic pain. The 5^th lambar (L5) DRG were obtaind after tight ligation of L5 an L6 spinal nerves for 7 days. Toluidine blue stain, immunohistochemistry for nNOS, eNOS, and iNOS, and NADPH-diaphorase histochemistry were performed. All specimens were observed under light microscope. nNOS expression was increased significantly in the cytoplasm of the DRG of ligated side at 7 day post-ligation (7D PL). eNOS expression was also increased in the blood vessel, a few neurons or satellite cells, and cell membrane of Schwann cells in the DRG of ligated side at 7D PL. iNOS expression was increased in a few blood vessels and cytoplasm of small neurons in the DRG of ligated side at 7D PL. NADPH-diaphorase expression was increased significantly in the neurons of the DRG of ligate side at 7D PL, but NADPH-diaphorase expressions were weak or absent in the blood vessel and Schwann cell in the contralateral DRG and the DRG of ligated side at 7D PL. It is suggested that NOS isoforms may be closely related to development and mantanence of neuropathic pain in order of nNOS>eNOS>iNOS.