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      • 간세포암과 감별이 어려웠던 악성중피종 치험 1예

        김선문,허원석,채경훈,강윤세,정재훈,김연수,박기오,문희석,이엄석,김석현,성재규,이병석,이헌영,신경숙,조준식,송인상,강대영 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2

        Malignant peritoneal mesothelioma is a rare neoplasm that arises from the mesothelium of a serosal cavity and is a rapidly fatal disease with a median survival of 4 to 12 months for untreated cases. Recently, we experienced a case with malignant peritoneal mesothelioma who was suspected hepatocelluar carcioma by abdominal CT scan and was confirmed by biopsy including immunohistochemical stain(calretinin) after surgery. We performed tumor excisions and wedge resection of the liver(segment Ⅷ)and inserted Tencoff catheter in abdominal cavity at 25th day of post-operation. We treated with intraperitoneal paclitaxel(25mg/m^(2)/day for 5 days) six courses monthly. She was well tolerable and is still living without any evidence of recurrence for 14th month of post-operation.

      • Gilbert 증후군에서 열량 제한 시험과 Phenobarbital 자극 시험의 의의(14예)

        이헌영,채경훈,정재훈,강윤세,김연수,문희석,박기오,이엄석,김선문,김석현,성재규,이병석,이강욱 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2

        Gilbert 증후군은 인구의 7%에서까지 나타날 수 있는 매우 흔한 증후군으로서 비진행성인 양성의 만성적 경과를 치하며, 간질환의 증상과 징후가 없는 경한 비포합형 고빌리루빈혈증이 특징인 일종의 체질적인 증상으로서 혈장 빌리루빈 농도에 대한 사춘기의 영향 때문에 10대와 20대에 자주 진단이 된다. 따라서 임상적인 중요성은 미약하지만 높은 빈도가 예상되는 점에 그 중요성이 부여되어야 할 것이다. 따라서 적정한 임상적 진단법으로 기왕에 소개된 열량제한 시험과 phenobarbital 유도 시험을 시행하고 이들의 진단적 가치를 알아보기 위하여 본 연구를 시행하였다. 1990년 7월부터 1999년 4월까지 충남대학교병원에 내원하여 HBsAg, IgG anti-HBc 및 anti-HCV가 음성이고, 간 초음파 스캔에서 이상이 없으며, 혈청 AST, ALT 및 AP가 정상인 비음주자에서 경한 비포합형 고빌리루빈혈증이 있는 14예의 환자들을 대상으로 ^(99m)Tc-DISID 스캔을 시행하였으며, 기저 치 총빌리루빈 및 포합형 빌리루빈 치를 측정한 다음에 하루에 400Kcal로 48시간동안 제한한 열량 제한 시험을 시행하였고, phenobarbital을 하루 60mg씩 5일간 투여한 후에도 각각 총빌리루빈과 포합형 빌리루빈 치를 검사하여 비포합형을 구하였다. 대상 환자들은 모두 14예로서 남자가 11예(78.6%)였고 여자가 3예(21.4%)여서 3.7:1로 남자에서 많았으며, 20대가 6예(42.9%), 30대가 역시 6예(42.9%) 및 40대가 2예(14.2%)로서 2,30대가 대부분(85.8%)이었다. 열량 제한 시험 후의 총빌리루빈 치, 비포합형 및 포합형 빌리루빈 치들은 평균 각각 5.5±2.7, 4.2±2.3 및 1.3±10mg/dL 로서, 시험 전 치들인 3.0±0.8, 2.2±0.8 및 0.7±0.4mg/dL 보다 유의하게(p=0.001, p=0.001, p=0.023) 상승하였다. 포합형 빌리루빈 치도 유의하게 상승하였으나 비포합형의 상승보다는 훨씬 낮아서 주로 비포합형이 증가하였다. phenobarbital 투여 중 설사가 발생하여 중단한 1예를 제외한 13예에서 열량 제한 시험 후에 상승하였던 총, 비포합형 및 포합형 빌리루빈 치가 phenobarbital 유도 시험후에는 2.0±1.1, 1.5±0.8 및 0.5±0.4mg/dL로서 열량 제한 시험 결과보다 유의하게 낮아졌고(p=0.00, p=0.000, p=0.001), 열량 제한 시험 전의 기초치들인 3.0±0.8, 2.2±0.8 및 0.7±0.4mg.dL 보다도 더욱 낮아졌으며 유의한 차이(p=0.001, p=0.02, p=0.005)를 나타내었다. 14예에서 시행한 ^(99m)-Tc DISIDA 스캔에서 9예(64.3%)가 정상이었고, 5예(35.7%)에서는 심장 및 신장으로의 간외 섭취가 3예였고, 60분까지 소장 배출이 없는 배설 지연 예와 담낭 수축 불량 예가 각각 1예 씩 발견되었다. Phenobarbital 투여시험에서 민감도가 열량제한시험에 비해 더 높았다(92.3%와 50.0%). Gilbert 증후군에서 1일 400 Kcal로 48시간의 열량제한 시험과 1일 60mg의 phenobarbital을 5일간 투여하는 유도 시험은 편리하고 유용한 임상적인 진단법으로 이용할 수 있다고 생각된다. 그러나 열량 제한 시험에서는 증가 기준의 통일이 필요하다고 유추되며 phenobarbital 유도 시험이 민감도가 더 높은 것으로 생각된다. Gilbert's syndrome is very frequent and benign chronic process characterized by mild, intermittent, unconjugated hyperbilirubinemia without any symptom and sign of liver disease. Previously intoduced caloric restriction test and phenobarbital stimulation test as two appropriate clinical tests had been examined and their diagnostic values were reevaluated. Fourteen patients with mild, persistent, unconjugated hyperbilirubinemia were included. Subsequently caloric restriction has been applicated by 400 Kcal/day for 48 hours and phenobarbital has been prescribed by 60 mg/day for 5 days. Therafter serum levels of total and direct bilirubin were measured. Most of the patients were third and fourth decade(85.8%) and male predominant. Each basal serum levels of total, indirect and direct bilirubin were 3.0±0.8, 2.2±0.8 and 0.7±0.4 mg/dL. After caloric restriction test, each levels were increased significantly to 5.5±2.7, 4.2±2.3 and 1.3±1.0 mg/dL(p=0.001, p=0.001, p=0.023). After phenobarbital stimulation test for 13 patients had been practiced, increased levels of each bilirubin after caloric restriction test were decreased significantly to 2.0±1.0, 1.5±0.8 and 0.5±0.4 mg/dL(p=0.000, p=0.000, p=0.001) and these levels were significantly lower than basal levels(p=0.001, p=0.02, p=0.005). The sensitivities of caloric restriction test were 85.7%, 50.0%, and 71.4%, 35.7%(1.0, 1.5 mg increase of total bilirubin and 1.0, 1.5 mg/dL increase of indirect bilirubin). The sensitivities of phenobarbital stimulation test were 93.2% and 92.3% at criteria of 1.5 mg/dL increase of total bilirubin and indirect bilirubin. On the diagnosis of Gilbert syndrome, caloric restriction test and phenobarbital stimulation test are convenient and useful diagnostic tools in clinical face. And also phenobarbital stimulation test has higher sensitivity than caloric restriction test. Furthermore, standardization of bilirubin increment would be necessary in caloric restriction test.

      • 자가 면역성 간염에서 자가항체의 발현 양상과 스테로이드 치료의 효과

        박기오,채경훈,허원석,강윤세,정재훈,김연수,문희석,이엄석,김선문,이병석,김남재,이헌영 충남대학교 의과대학 의학연구소 2003 충남의대잡지 Vol.30 No.1

        목적 : 최근 8년간 본원에서 경험한 자기면역성 간염의 혈액학적 특징과 스테로이드의 치료효과를 알아보고자 본 연구를 시행하였다. 방법 : 1996년 이후부터 2002년까지 충남대학교 병원에서 1999년 International Autoimmune Hepatitis Group(LAHG)에서 제시한 수정 진단 기준안에 의거하여 총 진단점수가 probable 이상에 해당하는 환자들을 대상으로 임상적 특징, 검사실 및 조직 소견과 스테로이드와 azathioprine의 치료 효과를 후향적으로 조사하였다. 결과 : 9예의 환자들은 여성이 78%, 40대가 많았고, 급성 발병이 33.3%, 피로감, 식욕부진 및 황달 등이 나타났으며, 항핵항체가 55.6%에서 그리고 항평활근 항체가 77.8%에서 양성이었고, 동반 질환은 류마치스 관절염이 2예, 특발성 혈소판 감소증 1예, Henoch-Sch nlein 자반증 및 그레브스병이 각각 1예 씩이었다. 7예에 대해서 스테로이드와 azathioprine으로 치료하여 생화학적 임상적 완해가 이루어져서 유효한 치료 결과를 얻었다. 결론 : 자가면역성 간염은 우리 나라에서 원인을 찾을 수 없는 만성 간질환의 경우, 특히 중년여성에서 다른 자가면역성 질환이 동반되었을 때 자가면역성 간염을 의심하여 자가항체 선별검사를 시행해 보아야 할 것으로 생각된다. 향후 다기관 연구를 통하여 좀 더 많은 증례를 모으고 전향적인 연구를 시행하여야 할 필요가 있다. Although autoimmune hepatitis(AIH) is common in western contries, some studties have been reported in Korea. The aim of the study was to analyze the clinical and histological features and expression pattern of the autoantibody and to examine clinical course of AIH in Korea. For 9 patients diagnosed as having AIH in our hospital since 1996, we reviewed medical records and analyzed clinical and laboratory findings retrospectively. They had a probable or definite AIH according to the revised criteria of International AIH group. The mean age was 42.7(21∼68)years and 7(77.8%) were female. Three of them(33.3%) showed acute hepatitis and five of them showed cirrhosis. Fatigue and anorexia were the most frequently complained symptoms(55.6%). Four patients(44.4%) were accompanied with other autoimmune diseases. Antinuclear antibody was detected in 5(55.6%) and anti-smooth muscle antibody was detected in 7(77.8%). Three patients showed focal picemeal necrosis and four showed periportal fibrosis. Among 7 patients who received immunosuppressive therapy, six patients(85.7%) showed initial response. and biochemical and serologic remission were achived in 5(71.4%) and 3(42.8%) patients, AIH seems to be partly responsible for chronic liver disease of unknown etiology in Korea. AIH should be suspected in female patients with unknown chronic liver disease of unknown etiolgy, especially accompanied with other autoimmune disease. The clinical characteristics of AIH may not be quite different from those of Western countries.

      • 히스토아크릴을 이용한 십이지장 궤양 출혈의 지혈 후 발생한 급성췌장염

        유재훈,문원,노지훈,구동영,조영화,김기수,윤준모 고신대학교 의과대학 2010 고신대학교 의과대학 학술지 Vol.25 No.1

        Upper gastrointestinal bleeding is common and potentially life threatening medical emergency. Emergency endoscopy is the first choice of diagnostic and treatment measure for patients with active upper gastrointestinal bleeding. Histoacryl should be considered a second-line treatment modality when conventional treatments were failed. However, it might cause several complications. Herein, we report a case of acute pancreatitis developed after histoacryl injection therapy for active duodenal ulcer bleeding. A 71-year-old man was admitted with melena and hematemesis. On emergency endoscopy, a 2 cm sized active ulcer with bleeding from an exposedvessel was seen at the duodenal bulb. Attempts to arrest the bleeding with hemoclipping and submucosal epinephrine injection were tried, but failed. We changed the method to endoscopic histoacryl injection, and obtained hemostasis immediately. A few hourslater, after successful hemostasis, patient complained diffuse abdominal pain. Ultrasonography revealed hyperechoic heterogenous diffuse pancreatic enlargement and right pararenal space fluid collection, this ultrasonographic findings and elevated serum pancreatic enzymes are compatible with acute pancreatitis.

      • KCI등재
      • KCI등재후보

        망간 노출 근로자의 정신 ·신경행동 양상에 영향을 주는 요인

        문영한,권기련,김은아,김규상,김양호,김재우,노재훈,양정선,진영우 大韓産業醫學會 1999 대한직업환경의학회지 Vol.11 No.2

        The risk posed to human health by environmental manganese (Mn) exposure is unknown. The purpose of this study is to establish if subclinical effects related to Mn exposure and examine the factors influencing psychoneurobehaviral outcomes of Mn exposed workers. This study involved 121 male workers of welding, alloy furnace, and manufacturing of welding stick. Study investigations include: a questionnaire covering demographic characteristics, job and exposure history and medical symptoms and conditions, monitoring workplace air for personal exposure to respirable and inhalable manganese, analysis of blood and urine samples, psychoneurobehaviral test(neurobehaviral core test battery(NCTB), signal change on magnetic resonance imaging (MRI) scans, minimental state and neurological examination). The main results of this study were as follows; 1. Preliminary results showed that, with increasing manganese exposure, neurobehaviral performance was poorer and signal change on MRI scans increased. 2. The results of examination revealed significant relations on psychoneurobehaviral outcomes (neurobehaviral performance, signal change on MRI scans, and neurological features). 3. In multiple regression, age and educational status as demographic variables and exposure level were significantly related to neurobehaviral test. Also exposure level and serum Mn concentrations were positively related to signal change on MRI scans and neurological features affected by Mn on the brain (signal change). 4. Psychoneurobehaviral outcomes from Mn exposure were related to neurobehaviral performance, signal change on MRI scans, neurological features and profile of mood states (POMS), and influenced positively with age, alcohol and smoking history, and duration of Mn exposure, negatively with educational status. Serum Mn concentrations in combination with brain MRI scans, and perhaps a battery of neurobehaviral tests, appear to be the best way to monitor excessive exposure to Mn. These results are consistent with our knowledge on Mn action on the brain and are similar to the type of neurobehaviral dysfunction. They suggest that there may be age, educational status, and life style (alcohol and drinking history) differences with chronic environmental exposure. These findings suggest further evaluation, particularly on relationships between Mn exposure, aging, and susceptibility factors.

      • KCI등재후보

        지역사회획득 폐렴의 치료지침 권고안

        송재훈,정기석,강문원,김도진,배현주,서지영,심태선,안중현,안철민,우준희,이남용,이동건,이미숙,이상무,이영선,이혁민,정두련,지역사회획득 폐렴 치료지침 제정위원회 대한감염학회 2009 감염과 화학요법 Vol.41 No.3

        A successful therapy of community-acquired pneumonia requires appropriate empirical antimicrobial therapy. Etiology and antimicrobial susceptibility of major pathogens of pneumonia can differ by country. Therefore, an ideal treatment guideline of community-acquired pneumonia should be based on the studies performed in each country. We developed a treatment guideline for community-acquired pneumonia in immunocompetent adults in Korea. This guideline was developed by the joint committee of the Korean Society for Chemotherapy, the Korean Society of Infectious Diseases, and the Korean Academy of Tuberculosis and Respiratory diseases.

      • 7개 대학 병원에서 조사한 지역사회 폐렴의 원인균

        정문현,김성민,강문원,최희정,정희진,이경원,한성우,송재훈,신형식,김의종,최강원,김민자,박승철,배현주,정윤섭,김준명,백경란,신완식,이규만,김양리 대한감염학회 1997 감염 Vol.29 No.5

        목 적 : 폐렴은 많이 발생하면서 사망률이 크게 줄지 않는 질환이며, 이를 적절히 치료하기 위해서는 원인균의 상대적 빈도, 기저 질환에 따른 변화, 항균제 내성률, 사망에 관련된 인자들을 알아야 한다. 원인균의 빈도는 지역마다 차이가 있고 국내에서는 항균제 내성률이 높아 지역사회에서 발생한 폐렴을 치료하기 위한 경험적 항균제 선택에 도움이 되기 위해 서울 소재 6개 대학 병원과 천안의 1개 대학 병원이 참여하여 위의 사항들에 대해 조사를 하였다. 방 법 : 1995년에 내과에 입원했던 16세 이상 환자를 대상으로 했다. 퇴원 진단명이 폐렴 또는 폐결핵인 병록지을 찾았고, 이중에서 병원 감염을 제외하였다. 특이도를 높이기 위해, 이들 중에서 호흡기 증상이 있고 발열이나 저체온이 있으면서 흉부 X-선에서 이상 음영이 있는 환자만을 대상으로 했다. 폐결핵은 위의 기준에 입원 초기에 항균제 치료를 하고 입원 7일 이후에야 항결핵제가 투여된 경우만을 폐렴의 원인균으로 하였다. 혈액 배양에서 양성, 객담에서 항상균이나 M. tuberculosis가 증명된 경우, 혈청학적으로 항체가가 4배 이상 증가된 경우, 조직에서 원인균이 진단된 경우는 확정(definitive) 원인균으로 하였고, 객담에서 배양된 균이 그람 도말과 일치할때, 항결액제에 대한 반응으로 진단한 폐결핵, 단일 항체가 양성이고 이에 대해 항균제를 사용했을 때는 가능(probable) 원인균으로 정의하였다. 다세균 감염균은 각각 다 른 원인균으로 처리하였다. 임상 조사와 함께 임상병리과에서 S. pneumoniae, H. influenzae, M. catarrhalis, mycoplasma, 항상균에 대해 검사 의뢰 건수, 배양 양성수, 항균제 감수성 결과를 조사하였다. 결 과 : 폐렴의 증례 정의에 부합하지 않은 135명과 폐결핵의 정의에 해당하지 않는 230명을 제외하고 남은 246명의 평균 나이는 58.2세이고 남성이 142명(58.2%) 이었고, 71%의 환자에서 기저 질환이 있었다. 진단 방법의 시행 횟수는 혈액 배양 77.6%, 혈청 검사 18.3%, 기관지경 검사는 4.1%였고, 세균의 항원 검사를 한 예는 없었다. 원인균이 밝혀진 예는 77명(31.3%)이었다. 다세균 감염이 4명에서 있었고, 원인균의 상대적 빈도는 결핵 20명(확정 17, 가능 3: 6개 병원 자료), 폐렴구균 18(확정8 가능 10)명과 폐렴구균이 아닌 Streptococcus 3명 (모두 확정), H. influenzae 11명(모두 가능), 그람음성간균 11명(확정 7, 가능 4) (K. pneumoniae 8건), Mycoplasma 5명(확정 1, 가능 4), S. aureus 4명(확정 2, 가능 2), mucormycosis 1명(확정)이었다. 평균 입원 기간은 19일이고, 중환자실 입원률과 인공 호흡기 사용율은 각각 18%와 9.3%였다. 사망률은 13.8%였고 사망까지 평균 기간은 14.6일 이었다. 다변량 분석에서 사망을 예측할 수 있는 인자는 저체온과 빈호흡이었다. 임상병리과에서 배양되었던 모든 폐렴구균의 Penicillin 내성률은 서울 3개 병원에서 82-88%, 천안에서 72%였다. 폐렴 환자의 혈액에서 배양된 7주는 모두 Penicillin에 감수성이 있었다. K. pneumoniae 8주 모두 cefotaxime과 gentamicin에 감수성을 보였다. 결 론 : 후향적 조사이고 병원마다 원인균 진단에 차이가 있지만, 원인이 밝혀진 경우에는 결핵과 폐렴균이 흔하였고, 무균 부위에서 배양된 폐렴구균의 항균제 내성률은 낮았다. 원인이 밝혀지지 않은 경우가 많고, 혈청검사로 진단되는 원인균이 드물며, 분리균주가 적어 항균제 내성 정도를 추정하기 어려워, 이를 밝히기 위한 전향적 조사가 필요하다. Background : Community-acquired pneumonia (CAP) is one of the leading causes of mortality and morbidity, but its management is still challenging. The limitation of diagnostic methods to identify etiologic agents rapidly make it necessary to use empiric antibiotics in almost all patients, and furthermore the discovery of new respiratory pathogens and the emergence of antibiotic-resistant organisms pose difficulties to the selection of an empiric regimen. To clarify the factors necessary for the optimal choice of empirical antibiotics, such as the frequency of etiologic agents, the attributable rates to death and antimicrobial resistance rates in the community, six university hospitals in Seoul and one university hospital in Cheonan were participating in this study. Methods : medical records of adults (>15 years of age) hospitalized for CAP or pulmonary tuberculosis between April 1995 and March 1996, were reviewed. Patients who satisfied all of the following criteria were included in the study: (1) fever or hypothermia; (2) respiratory symptoms; and (3) pulmonary infiltrates on chest roentgenogram. To exclude cases of pulmonary tuberculosis whose roentgenographic features were so typical that it could be easily differentiated from conventional pneumonia, two additional criteria were required for inclusion: antibiotic treatment during the first week of hospital admission and initiation of anti-tuberculosis medications thereafter. Organisms isolated from sterile body sites, acid-fast bacilli or Mycobacterium tuberculosis isolated from sputum, pathogens diagnosed by a 4-fold rising titer of antibodies to “atypical”pathogens, or pathogens revealed by histopathology were defined as definitive cause of pneumonia; isolates from sputum withcompatible Gram stain, pathogens diagnosed by a single diagnostic titer plus use of a specific antimicrobial agent, or tuberculosis diagnosed by clinucal response to anti- tuberculosis medications were considered probable cause of pneumonia. The records of the clinical microbiology were reviewed for isolates of S. pneumoniae, H. influenzae, M. catarrhalis, Mycobacterium or acid-fast bacilli, and Mycoplasma. Then the frequency of these agents, antimicrobial resistance rates of resiratory pathogens from all body sites, and their clinical significance were evaluated. Results: After excluding 365 patients (230 with pulmonary tuberculosis and 135 with CAP) who were screened for inclusion but did not meet the inclusion criteria,246 persons were enrolled in this study. Their mean age was 58.2 years old with slight male predominance (58.2%), and 171(71%) patients had underlying illnesses. Blood cultures were performed on 191 (77.6%) patients and serologic tests on 44(18.3%) patients. The etiologic agents were identified in 31.3%, and the list of individual agents, in decreasing order, was pulmonary tuberculosis (17 definite and 3 probable: data of six hospitals), S. pneumoniae (8 definite and 10 probable), non-pneumococci (3 definite), aerobic gram-negative bacilli (7 definite and 4 probable), Haemophilus spp. (11 probable), mycoplasma (1 definite and 4 probable), polymicrobial infections (2 definite and 2 probable: E. coli and S. agalactiae, M. tuberculosis and S. aureus, S. pneumoniae and H. influenzae and A. baumannii and K. pneumonias), S. aureus (2 definite and 2 probable) , and mucormycosis (1 definite). Among gram-negative bacilli, K. pneumoniae was the most common agent (8isolates). therates of admission to the intensive care unitand of using assisted ventilation were 18% and 9.3%, respectively. The mortality was 13.8% and logistic regression analysis showed that hypothermia and tachypnea were associated with death. Hospital stay averaged 19 days. Susceptible rates of S. pneumoniae isolated from all body sites to penicillin ranged from 8% to 28% but seven isolated from blood of patients with pneumonia were susceptible to penicillin. Also all 8 isolated of k> pneumoniae from patients with pneumonia were susceptible to cefotaxime and gentamicin. Conclusion: In Korea, in addition to S. pneumoniae, M. tuberculosis is an important agent causing community-acquired pneumonia. The low incidence of etiologic diagnosis is probably related to infrequent requesting of test "atypical" pathogens and does not represent the true incidence of infections by "atypical" pathogens, which well be answered by a prospective study. The antimicrobial resistance rates of major respiratory pathogens from sterile body sites are low, however, because of a small number of the isolates this result needs confirmation by a nationwide surveillance of antimicrobial resistance.

      • 白花蛇舌草 메탄올 抽出物의 抗腫瘍 效果 및 抗癌 棋戰에 關한 硏究

        魯勳政,文九,文錫哉,元秦熹,文永昊,朴來佶 대한한방종양학회 2000 대한한방종양학회지 Vol.6 No.1

        Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to 0.4㎍) and periods (6 to 30 hr) of H_2O and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with 200㎍/ml of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with 200㎍/ml of each extract for 16hr.Then, cells were treated with various doses of each extract for 12 hr and 100㎍/ml of methanol extract for various periods. Lysate from the cells used to measure the activity of caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively, Cells were treated with 200㎍/ml of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with 32^p-ATP and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by suing Phosphoimage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of NF-kB was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at CO_2 incubator for 6 days. The number of colony was cunted under light microscopy (×100). Results: The death of HL_60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL_60 cells. In addition, it was shown nucleus chromatin condensation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspaxe 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, NF-kB was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator NF-κB, In addition, our results also suggest that methanol exthanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

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