저산소성 허혈성 뇌손상을 유발시킨 어린 백서에서 Myeloperoxidase 측정 검사

양혜정(Hae Joung Yang),피대훈(Dae Hun Pee),임지혜(Ji Hye Lim),최병민(Byung Min Choi),이기형(Kee Hyoung Lee),은백린(Baik-Lin Eun) 대한소아신경학회 2002 대한소아신경학회지 Vol.10 No.2

목 적 : 예전에는 뇌경색 부위에서 발견되는 백혈구를 단지 뇌손상에 뒤따르는 생리적인 반응으로만 생각하였으나 최근 10년 사이 재관류 손상에서도 백혈구에 의한 염증 작용이 중요한 역할을 한다고 알려지면서 이들의 역할에 대한 연구가 새롭게 이루어지고 있다. PML의 존재는 세포질내 과립에서 분비되는 MPO를 생화학적으로 측정함으로서 증명할 수 있는데 저자들은 미성숙 뇌에서 저산소성 허혈증에 의한 뇌손상이 발생할 때 백혈구가 침윤되는 과정을 MPO 측정 검사를 통하여 관찰하였고, P와 L-selectin 억제제인 Fucoidin을 투여한 후 MPO의 변화를 확인하였다. 방 법 : 생후 7일된 어린 흰쥐를 사용하여 우측총 경동맥을 전기 응고시켜 자르고 8% 산소에 노출시켰다. 저산소 노출 후 회복 시간에 따라 동물을 희생시키고 뇌를 추출하여 뇌조직 1 g당 10mL의 20 mM potassium Phosphate buffer(pH 7.4)를 첨가하여 50초간 분쇄하였다. 각각의 균질회된 조직 샘플들은 4℃에서 20분 동안 원심 분리하여 상층액을 제거한 후, 침전물은 처음 조직량에 따라 조직 1 g당 10 mL의 0.5% cetylditrimethylethyl ammonium bromide(wt/vol)가 첨가된 50 mM potassium phosphate buffer(pH 6.0)를 첨가하였다. Vortex를 이용하여 buffer와 침전물을 잘 섞고 60℃수조에서 120분 동안 방치하였다. 그 후 각 조직 샘플들을 4 watts에서 음파처리하고 4℃에서 15분 동안 원심 분리를 시행하여 상층액을 새로운 tube에 옮겼다. MPO 측정검사를 위한 MPO reaction buffer는 50 mM potassium phosphate buffer(pH 6.0) 100 mL에 0.53 mM odianisidine edihydrochlorde와 0.0005% HO를 섞어 만들었다. MPO reaction solution 2.9 mL에 각각의 MPO sample 0.1 mL을 첨가하여 460 nm의 파장에서 5분 동안 MPO에 다른 흡광도 차이를 관찰하였다. 결 과 : 조직내 백혈구 침윤의 지표로 측정한 MPO 활성도는 대조군에서는 미미하였으나 총 경동백을 절단한 우측 대뇌군에서는 허혈 및 저산소증 유발 후 8시간째부터 현저히 증가되기 시작하여 24시간 경과 후 발현이 가장 높았으며, Fucoidin 50 mg/kg으로 전처치한 약물 실험군에서는 MPO 활성도가 현저히 감소하였다. 결 론 : 백혈구는 미성숙 뇌의 저산소성 허혈증에 의한 뇌손상에서 중요한 매개체 역할을 하며, 백혈구의 생화학적 활성도를 나타내는 MPO 활성도는 저산소성 허혈성 뇌손상의 정도를 측정하는 지표로 이용될 수 있다고 생각된다. Purpose : Neutrophils found around an infarcted area in the brain was once considered as only the physiologic response following the brain injury, but recent studies have shown that inflammatory responses by neutrophils play an important role in the reperfusion injury. The presence of ploymorphonuclear leukocytes(PML) is proven by biochemical assay of myeloperoxidase(MPO) Secreted in the cytoplasmic granules. We observed the process of PML infiltration on hypoxic-ischemic brain injury of immature rats by the assay of MPO activity and changes of the MPO activity after the administration of fucoidin, inhibitor of P- and L-selectin. Methods : We used a well characterized model of the brains of 7 day-old-rats, which had unilateral hypoxic and ischemic injuries(HI). Those injuries were induced by unilateral carotid artery ligation followed by timed exposure to hypoxic inspiratory gas mixture(8% O). MPO activity was measured in the brain tissue homogenates of HI rats(n=18) at 0, 2, 8, 24 and 48 hrs and in rats that received fucoidin immediately before and again after hypoxia(50 mg/kg, n=6) at 8 and 24 hrs. Controls(n=2) were rats with neither hypoxia nor ischemia. The brain samples were homogenized in 20 mM potassium phosphate buffer(pH 7.4) for 50 secs. The homogenate was centrifuged at 14,000 g at 4℃ for 15 mins and the supernatant was discarded. The tissue was pulverized, weighed, and suspended in 1 mL of 50 mM potassium phosphate buffer solution(pH 6.0) contatining 0.5% cetylditrimethylammonium bromide(wt/vol). The tissue was sonicated and centrifuged at 10,000 g for 15 mins. 200 µL of the supernatant was mixed with 1 mL of 50 mM potassium phosphate buffer solution(pH 6.0) containing 10 μL of 1.325 mM o-dianisidine hydrochloride and 170 µL of 3% hydrogen peroxide(vol/vol). Changes in absorbance at 460 nm were measured for 5 mins by using microplate reader. One unit of MPO activity was defined as that degrading 1 µmol peroxide/min at 25℃, and the result was expressed as units of MPO/100 gm tissue. Results : In HI rats, MPO activity increased at 2 hrs after HI and peaked at 24 hrs in the right hemisphere. In rats with fucoidin treatment immediately before and again after hypoxia, the MPO activity significantly decreased in both hemispheres compared with HI rats(P<0.05). MPO activity in the tissue of control rats was insignificant. Conclusion : The dynamic changes of the MPO activity suggest the important role of PMN on hypoxic-ischemic brain injuries in immature rats. MPO activity could be used as an index of the severity of injuries of hypoxic-ischemic brains.

위절제술 환자의 표준진료지침 개발 및 적용 효과

김은희,김철규,이순교,김순덕,이혜옥,권정순,이경미,이민미,심순미,유용만,신종식,강은희,이상일,김병식,오성태,육정환,박수길 한국의료QA학회 2003 한국의료질향상학회지 Vol.10 No.2

Background : Gastric cancer is the most common malignant tumor in Korea. surgical operation is one of the major treatment modalities for gastric cancer patients. Therefore, gastrectomy is one of the most common procedures in General Surgery. There were variation in length of hospital stay and medical treatment for gastrectomy between three surgeons at Asan Medical Center. Clinical pathways have received considerable attention as a tool for recucing the medical practice variation, increasing the efficiency of care process, and improving the quality of care. The aim of this study was to evaluate the effect of a clinical pathway for gastrectomy in gastric cancer patients. Methods : The clinical pathway for gastrectomy was developed and implemented by a multidisciplinary group in Asan Medical Center. A computerized clinical pathway program was developed and revised after a pilot test. A total of 145 patients underwent gastrectomy by three surgeons at Asan Medical Center. We compared the length of hospital stay, patient satisfaction, and unplanned readmission rate between the pre-pathway group(n=67) and the post-pathway group(n=78). We also investigated the degree of satisfaction among the physicians and nurses who were main end-users of the clinical pathway. Results : The clinical pathway was applied to all target patients. The average length of hospital stay was shortened from 12.7days to 10.6days(p<0.01). The degree of patient satisfaction with the care process changed from 90.3% to 89.2% after the implementation of the clinical pathway, but the difference was of satistically significant(p=0.761). Unplanned readmission rate was 2.9% in the pre-pathway group. More than 90% of physicians and nurses answered that the clinical pathway had been a useful tool in their medical practice. Conclusions : The findings of the study demonstrated that implementation of the clinical pathway for gastrectomy produced substantial reduction in the length of hospital stay while improving the quality of patient outcomes. The computerized clinical pathway program can be used as one of the powerful patient management tools for reducing the practice variations and increasing the efficiency of care process in Korea hospital settings.

수준별 수영 참여자의 동기유형 분석

배병석,정재은,육동원,김지수,김민현 한국스포츠리서치 2005 한국 스포츠 리서치 Vol.16 No.2

The purpose of this study was to examine motivation type of swimmers by competency level. A total of 262 swimmers were composed of 111 beginners, 58 inter-mediates and 93 advancers participated in the study. For the study, duration of participation, frequency of participation and the level of sport performance of participants were set as the independent variable, while motivation type was the dependent variable. The study used analysis methods such as frequency analysis, t-test, one-way ANOVA between the independent and dependent variables. A significant difference was found after conducting an analysis on the difference in motivation type by duration of participation. The analysis on the difference in motivation type by frequency of participation showed that a group that swam three to five times per week recorded higher marks in terms of physical condition and social life factors than that swam two to three times per week. The analysis on the difference in the motivation type by level of sport performance of participants showed a significant difference in development of skills, enjoyment, achievement, physical condition. social life. external ostentation and amotivation factors.

정상상피세포(HaCat)와 자궁경부 암세포(SiHa)에서 GeneFishing^(TM) PCR technique을 이용한 유전자 발현의 변화

김병훈,배수미,서민제,김용완,이정웅,김용욱,이준모,남궁성은,김종국,안웅식 대한부인종양 콜포스코피학회 2003 Journal of Gynecologic Oncology Vol.14 No.4

목적 : 본 실험의 목적은 정상상피 세포와 자궁경부 암세포 사이에서 유전자의 발현 차이를 조사하였다. 연구 방법 : 정상상피 세포(HaCat)와 자궁암 세포(SiHa)를 사용하였으며, 두 세포 간에 유전자 발현 차이를 GeneFish^(TM) PCR을 이용하여 알아보았으며, BLAST serach를 통해 분석하였다. 결과 : 정상상피 세포와 자궁암 세포 비교 결과, 자궁암 세포에서 S1-2-2와 S5-1을 포함한 25개의 유전자가 발현이 증가하였고, 24개의 유전자가 감소하였다. 결론 : GeneFishing^(TM) PCR기법은 유전자의 발현 변화를 확인하는데 있어서 아주 민감하고 효과적인 방법이다. 우리는 정상상피 세포와 자궁경부 암세포에서 다르게 발현하는 유전자를 찾을 수 있었고, 앞으로는, 종양의 발생과 진행과정에 관여하는 유전자를 더 탐지하고 해당 유전자의 기능을 연구할 필요가 있다고 생각된다. Objective : The purpose of this study is investigated the differentially expressed genes between normal and cervical cancer cell line. Methods : We used normal human keratinocyte (HaCaT) as a control and HPV-16 positive cervical cancer (SiHa) cell line. Two cell lines were studied differential expressed genes by using GeneFishing^(TM) PCR and analyzed with BLAST search. Results : As compared with normal, cervical cancer cell line was showed 25 up-regulated genes including the S1-2-2, S5-1 and 24 down-regulated genes. Conclusion : GeneFish^(TM) PCR test is very sensitive and effective method for detection of changed gene expression. We could search differentially expressed genes between normal and cervical cancer cell line. In the future, we need to research various genes function to participate in the process of tumor development and progression.

Skin entrance dose for digital and film radiography in Korean dental schools

Cho, Eun-Sang,Choi, Kun-Ho,Kim, Min-Gyu,Lim, Hoi-Jeong,Yoon, Suk-Ja,,Kang, Byung-Cheol 대한구강악안면방사선학회 2005 Imaging Science in Dentistry Vol.35 No.4

Purpose : This study was aimed to compare skin entrance dose of digital radiography with that of film radiography and to show the dose reduction achievement with digital systems at 11 dental schools in Korea. Materials and Methods : Forty six intraoral radiographic systems in 11 dental schools were included in this study. Digital sensors were used in 33 systems and film was used in 13 systems. Researchers and the volunteer visited 11 dental schools in Korea. Researchers asked the radiologic technician (s) at each school to set the exposure parameters and aiming the x-ray tube for the periapical view of the mandibular molar of the volunteer. The skin entrance doses were measured at the same exposure parameters and distance by the technician for each system with a dosimeter (Multi-O-Meter : Unfors instruments, Billdal, Sweden). Results : The median dose was 491.2 μGy for digital radiography and 1,205.0 μGy for film radiography. The skin entrance dose digital radiography was significantly lower than that of film radiography (p<0.05). Conclusion : Fifty-nine percent skin entrance dose reduction with digital periapical radiography was achieved over the film radiography in Korean dental schools.

Rhizopus nigricans Ehrenberg의 Histones에 관한 연구

이은영,민병례 상명대학교 기초과학연구소 1989 기초과학연구 Vol.3 No.-

하등한 진핵생물인 Rhizopus nigricans Ehrenberg로 부터 basic chromosomal protein을 분리하여 고등한 진핵생물의 histones 과 비교하였다. 대수기의 균사체로부터 핵을 분리하였으며 sucrose density gradient centrifugation의 방법으로 purify하였다. Chromatin으로부터 basic protein의 추출을 위하여 0.2N HCl과 최종농도 0.4N의 H2SO4와 6부피의 acetone을 사용하였다. 15% acrylamide acetic acid-urea gel electrophoresis 결과 calf thymus histones과 동일한 mobility를 나타내는 4개의 band가 나타났다. Band Ⅰ protein은 다른 진핵생물의 histones처럼 heterogenetiy를 나타냈으며, Band Ⅱ와 Ⅲ는 calf thymus H2A, H2B 그리고 H3와 동일한 mobility를 보여주었다. Band Ⅳ 역시 calf thymus histone H4와 동일한 mobility를 나타내었다. 이러한 실험결과를 근거로 하여 볼때 Rhizopus nigricans는 고등 진핵생물과 매우 유사한 basic histone proteins을 포함하고 있다고 추정 할 수 있었다. From Rhizopus nigricuns Ehrenberg, the lower eukaryote, basic chromosomal proteins were isolated and compared with higher eukaryotic histones. Nuclei were isolated from mycelia at exponential stage and purified with low RCF and sucrose density gradient centrifugation. For the extraction of basic proteins from chromatin, 0.2N HCl, H₂SO₄, of which final concentration is 0.4N, and 6 volumes of acetone were used. As a results of 15% acrylamide acetic acid - urea gel electrophoresis of fungal basic proteins, there appeared 4 bands showing identical mobility to those of calf thymus histones. Band I protein exhibited heterogeneity as any other eukaryotes did. Band Ⅱ and m showed the patterns identical or similar to those of calf thymus histone H2A, H2B and H3. And the mobility of Band Ⅳ is identical to calf thymus histone H4. On the basis of the results obtained through this experiment, it may be stated that Rhizopus nigricuns contains basic histone proteins which are very similar to higher eukaryotic histones.

오차드그래스(Dactylis glomerata L.)의 품종에 따른 종자유래의 캘러스 형성률과, 캘러스 크기 및 식물체 재분화 효율의 비교

배은경,이인애,김기용,이병현,손대영,이효신,정민섭,조진기 Plant molecular biology and biotechnology research 2002 Plant molecular biology and biotechnology research Vol.2002 No.-

Comparison results of callus formation ratio from seed explants, callus sizes, regeneration ratios from callus and regeneration efficiency [calculated by following formular; callus formation ratio(%) × regeneration ratio(%)/100]for 27 ochardgrass (Dactylis glomerata L.)varieties imported and Hapsung 2 developed in Korea are as follows; 1. Among ochardgrass varieties showing more than 50% callus formation ratios, the descending order of callus formation ratio after bedding the seed explants for 4 weeks was 93>Sparta>Pizza>Condor>Lidaglo>Glorus>Hapsung2>Frode. 2. The callus sizes after bedding for 4 weeks were in the range of φ 0.43cm~4.2cm in which there was 10 times size difference between the largest one and the smallest one but most of them were between φ2.5cm~4cm. 3. The regeneration ratio from callus among varieties were in the range of 0~36% and descending order of the upper 6 varieties was Plano>Akimidori>Justus>Lidacta>Currie>Hall mark. 4. The regeneration efficiency which is calculated by the ratios of regeneration from seed explant numbers was between 0 to 17.4% among which Justus showed the highest value in the 4-week treatment. 5. The correlation between callus formation ratios and the callus sizes, callus formation ratios and regeneration efficiency, and callus sizes and regeneration efficiency were r=0.5765, r=0.6365 and r=0.6246, respectively in 4-week callus and all the correlations were significant on the 1% level. 6. In 6-week callus, the descending order callus formation ratios from seed sxplants for the best 6 varieties was Condor>Sparta>93M>Justus>Potomac>Lidaglo>Frode. 7. The callus sizes formed were between φ1.5~5.7cm in which Sparta, the largest one of φ5.7cm was five times larger than the smallest one. The callus size of the control variety. Hapsung 2 was φ3.8cm, which belonged to larger size. 8. Regeneration ratio showed a great deviation among varieties from 6-week old calli by showing from 0% to 100% in which all the calli were regenerated in Plano while no callus was regenerated in Juno, 9. The range of regeneration effciency was between 0~28% among varieties in which the values from 6-week callus treatment were larger than those from 4-week callus treatment. Especially, the value of Potomac in 6-week was 3 times larger than that in 4-week. 10. The correlation between callus formation ratios and the callus sizes, callus formation ratios and regeneration efficiency, and callus sizes and regeneration effciency were r=0.8369, r=0.6683 and r=0.5937, respectively in 6-week callus, and all the correlations were significant on the 1% level.

Rhizopus nigricans Histones 의 SDS-Polyacrylamide Gel Electrophoresis 에 의한 분리

이은영,민병례 상명대학교 기초과학연구소 1990 기초과학연구 Vol.4 No.-

Rhizopus nigricans의 균사체로부터 핵을 분리하고 핵내에 포함되어 있는 basic chromosomal proteins을 정제하였으며, 고등진핵생물 histones과의 유사성을 알아보기 위하여 sodium dodecyl sulfate polyacrylamide gel electrophresis를 실시하였다. Proteins의 electrophoretic mobility와 분자량의 기준으로 calf thymus histones을 사용하였으며 그들이 가장 선명하게 분리되는 조건과 동일한 조건으로 R. nigricans로부터 분리한 proteins을 전기영동하였다. 그 결과, 균류중에서도 하등한 R. nigricans로부터 분리된 basic proteins이 고등진핵생물 histones의 5 fractions중 H2A, H2B, H3, H4와 mobility가 동일하거나 매우 유사한 basic histone proteins을 포함하는 것을 확인하였다. H1으로 간주되는 protein은 3개의 bands로 나타나 heterogeneity가 큰 것으로 사료되나 이를 확인할 수 있는 연구는 계속 진행되어야 할 것이다. Basic chromosomal proteins were purified from Rhizopus nigricans. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was achieved to compare the similarity between purfied proteins and higher eukaryotic histones. Calf thymus histones were used as a standard for electrophoretic mobility and molecular weight of histones. Electrophoresis of proteins from R. nigricans was carried out with the same condition as standard can be separated clearly. On the basis of these comparisons, it appeared that ,R. nigricans contains basic histone proteins which are identical or very similar with higher eukaryotic histone H2A, H2B, H3 and H4. And the proteins regarded as H1 are thought to be very heterogeneous as any other eukaryotes. But heterogeneity of H1 is remained to be discussed.

정신분열증환자에서 Haloperidol과 Reduced Haloperidol 사이의 상호변환에 있어 Haloperidol Reductase에 관한 연구

김민정,정은기,오동열,김이영,양병환 大韓神經精神醫學會 1994 신경정신의학 Vol.33 No.5

Haloperidol is metabolized through reductation by the enzyme haloperidol reductase at the benzylic ketone to form an alcohol metabolite, known as reduced haloperidol. The basic and clinical pharmacology of reduced haloperidol and also haloperidol reductase is the main subject of haloperidol study, as the reduced metabolite can be converted back to the parent drug which has some neuroleptic effect. This study was undertaken to investigate correlations between haloperiol reductase activity in RBC and reduced haloperidol formation, also the difference of haloperidol reductase activity between responding and nonresponding schizophrenic patient. The results were as follows : 1) The haloperidol reductase activity of schizophrenic patients in RBC at 2 weeks was 12.8±4.3ng/30 min/200ul RBC, 16.0±11.9 at 4 weeks and 22.3±71 in control group. There was statistically significant difference of haloperidol reductase activity in RBC between schizophrenic patient and control group at 2 weeks but no difference at 4 weeks. 2) We divided the patient into responding and nonresponding group by the percentage of improvement in total BPRS score as more or less than 31% respectively and there was no significant difference of haloperidol reductase activity in RBC among two groups. 3)) There was no significant relationship between haloperidol dose of administration and haloperidol reductase activity in RBC. Considering the above results, haloperidol reductase activity in schizophrenic patients showed different results according to the duration of drug administration, so the study which deal with the factors, influencing the activity of haloperidol reductase, should be undertaken in the future.

Phenazine Methosulfate의 Superoxide Radical 生成作用

李民化,安鳳煥,李起榮,朴昞柱,梁誠烈,薛鍾銀 全南大醫大 1984 전남의대학술지 Vol.21 No.2