Breeding of Tetraploid in Platycodon grandiflorum (Jacq.)A. DC. by Colchicine treatment

Kim,Ik-Hwan,Kim,Hag-Hyun,Hong,Eui-Yon,Yun,Jong-Sun,Yun,Tae,Hwang,Ju-Kwang,Lee,Cheol-Hee 한국자원식물학회 2003 Plant Resources Vol.6 No.3

Present studies were carried out to produce tetraploid plants by colchicine treatment using seeds, seedlings and shoot tips of Platycodon grandiflorum in Campanulaceae. The most successful colchicine treatment for tetraploid production in P. grandiflorum was soaking treatment using 0.01 and 0.5% colchicine solution for 1 hour and 12 hours, respectively. Morphological characteristics of both diploid and tetraploid were similar, but tetraploid plants had more leaves. Compared to diploid, tetraploid had the larger stomata, but less number of stomata. Fresh weight of tetraploids was 20∼40% heavier than that of diploid.

京畿道 廣州郡 및 富川郡에 있어서의 肺디스토마의 疫學的 調査

尹德鎭,金元撤,全世鍾,李俊鎬,金燉均,尹燾善,南相德 대한의학협회 1965 대한의학협회지 Vol.8 No.9

항암제 처리한 백혈병 세포주에서의 Apoptosis 발현

김삼용,윤소현,김현수,김종숙,윤환중,김진경,조덕연 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

The bcl-2 proto-oncogene encodes a 26 kD protein that promotes cell survival by blocking apoptosis. The bax protein is a member of the bcl-2 familly, now known to form heterodimers with the bcl-2 protein. The ratio of bax to bcl-2 is be critical in determining the fate of the cell in response to stimuli that can induce apoptosis. Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro with Topo I inhibitory action. In the present study, the relationship between apoptosis and the apoptosis related proteins, bcl-2 and bax were investigated in human leukemic cell lines HL-60, U-937 and CEM-CM3. All anticancer drugs(adriamycin, etoposide, camptothecin, SB-31) induced extensive apoptosis in HL-60, U-937 cells and CEM-CM3 cells. The expression of bcl-2 and bax protein were determined in cell lines by western blotting before and after incubation with anticancer drugs at different time points. 1) In HL-60 or U-937 cell lines, down regulation of bcl-2 and up-regulation of bax were found after incubation with ADR, VP-16 or camptothecin. 2) In HL-60 or U-937 cell lines, no significant change in bcl-2 or bax protein expression resulted after incubation with SB-31. 3) In CEM-CM3 cells, virtually no change was noted in bcl-2/bax expression after incubation with ADR, VP-16, camptothecin or SB-31. It is suggested that different leukemic cell lines use different pathways of apoptosis activation and a given cell may utilize different pathways of apoptosis activation in response to different cytotoxic agents.

한국인 갑상선 수질암 환자에서 RET 원종양유전자 점돌연변이 양상

김형훈,김현진,정윤재,민용기,이명식,이문규,김광원,기창석,김종원,정재훈 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.4

연구배경: 갑상선 수질암의 25∼30%는 유전성으로 발현되는데, MEN 2A, MEN 2B 또는 가족성 수질암의 형태로 나타난다. RET 원종양유전자의 점돌연변이가 유전성 갑상선 수질암의 발생에 중요한 역할을 하므로, 진단 당시의 연령이나 가족력 유무에 관계없이 모든 갑상선 수질암 환자나 또는 RET 변이가 발견된 수질암 환자의 가족 구성원들에서 RET 변이 검색을 하여야 한다. 또한 일부 문헌에서 RET 변이의 양상에따라 임상상이 다르게 표현됨이 보고된 바 있다. 이에 저자들은 한국인 갑상선 수질암 환자에서 RET 원종양유전자의 점돌연변이의 양성률을 알아보고, 변이 양상에 따른 임상상의 차이가 있는지를 알아보고자 본 연구를 시행하였다. 방법: 치근 7년간 본원에서 갑상선절제술을 통해 갑상선 수질암으로 진단받은 29예에서 RET 원종양유전자의 점돌연변이 검사를 시행하였다. 29예의 평균연령은 39세(20∼60세)이었고, 남자 7예, 여자 22예 이었다. 이들의 말초혈액에서 genomic DNA를 분리하고, 특이 시발차를 이용하여 RET 원종양유전자의 exon10, 11, 13, 14, 16부위를 증폭하였다. 증폭된 부위를 자동염기서열분석기를 이용하여 직접 분석하였다. 양성으로 나온 경우는 모든 가족 구성원을 대상으로 RET 변이 유무를 검색하였다. 결과: 대상 환자 29예 중 9예 (31%)에서 RET 원종양유전자 점돌연변이가 발견되었다 RET 변이가 발견된 9예 (남자 3예, 여자 6예)의 평균 연령은 33세 (20∼51세)로 RET 변이가 발견되지 않은 20예의 평균연령 42세(24∼60세)보다 의미 있게 적었다. RET 변이가 발견된 9예 중 MEN 2A가 5예, 가족성 수질암이 1예, 그리고 산발성 수질암이 3예이었고, MEN 2B는단 1예도 진단되지 않았다. MEN 2A 5예 중 4예는 exon 11의 codon 634번(C634R 2예, C634Y 2예)에서, 그리고 나머지 1예는exon 10의 codon 618번 (C618R)에서 변이가 각각 발견되었다. 가족성 수질암 1아는 codon 634번(C634W)에서, 산발성 수질암 3예도 모두 codon 634번 (C634y 2예, C634s 1예)에서 각각 변이가 발견되었다. RET변이 양상 또는 위치에 따른 임상상의 차이는 발견할 수 없었다. 결론: 갑상선 수질암 환자 31%에서 RET 원종양유전자의 점돌연변이를 발견할 수 있었다. 유전성 수질암 중 가족성 수질암 1예를 제외하고 나머지 5예는MEN 2A이었다. 저자들의 5예와 지금까지 국내에서 보고 된 7예를 합친 국내 MEN 2A 12예 중 75% (9/12)는 exon 11의codon 634번(C634R 4예, C634y 4예, C634w 1예)에서, 그리고 나머지 25% (3/12)는 exon 10의 codon618번(C618R 2예, C618s 1예)에서 변이가 발견되었다. 국내에서는 codon 634과 codon 618 두 곳에만 국한된 양상이었고, codon 634에서의 C634R 변이는 1/3에서만 나타났다. 비록 본 연구에서는 제한된 환자 수 때문에 변이 양상과 임상상의 관계를 규명할 수 없었지만, 향후 많은 수의 환자를 대상으로 전향적인 연구를 시행하여 genotype-phenotype 관계 규명을 하는 것이 필요하다. Background: Medullary thyroid carcinomas (MTC) have been reported as hereditary in about 25 -30% of cases. The identification of germline mutation in RET proto-oncogene is important in the diagnosis of hereditary MTC, and occurs in three forms: MEN 2A, MEN 2B and familial MTC (FMTC). To evaluate the prevalence of the relationship of RET proto-oncogene mutation and genotype-phenotype was studied in Korean patients with MTC. Methods: Genomic DNA was obtained from 29 patients, with MTC, who underwent a total thyroidectomy, between 1997 and 2003, at the Samsung Medical Center. There were 7 male and 22 female patients, with an average age of 39, ranging from 20 to 60 years. Exon 10, 11, 13, 14 and 16 of the RET proto-oncogene were amplified, with specific primers, using PCR. A sequencing analysis was performed on the PCR product using an automatic sequencing analyzer. Results: Nine of the 29 patients (31%) were identified as having RET mutations. The average age of these 9 patients was 33 years, ranging from 20 to 51, with a female to male ratio of 2. Five patients had MEN 2A and one had FMTC, with the other 3 thought to have non-hereditary (sporadic) MTC. The 4 patients with MEN 2A had RET mutations on codon 634 of exon 11 (2 patients, C634R; 2 patients, C634Y) and the other patient on codon 618 of exon 10 (C618R). One patient with FMTC had a mutation on codon 634 (C634W). Three patients with sporadic MTC had RET mutations on codon 634 (2 patients, C634Y; 1 patient, C634S). However, no genotype- phenotype relationship could be found, due to the limited number of patients. Conclusion: Thirty-one percent (9/29) of the patients with MTC had RET proto-oncogene mutations. Three-quarters (9/12) of the Korean patients with MEN 2A, including another 7 patients reported in 3 papers in Korea, had RET mutations on codon 634 of exon 11 (4 patients, C634R; 4 patients, C634Y; 1 patient, C634W), but a quarter (3/12) had mutations on codon 618 of exon 10 (2 patients, C618R; 1 patient, C618S). Although no relations could be found between the genotypes and phenotypes, extensive prospective studies will be required to verify this (J Kor SOC Endocrinol 18:360-370, 2003).

기종성 방광염을 동반한 기종성 신우신염 1예

김선두,김순제,이길도,김정수,김지훈,윤호상,송종오,문언수 대한노인병학회 2000 Annals of geriatric medicine and research Vol.4 No.3

Callus distraction method를 이용한 하악골 신장술 : 계단골절단술식의 적용

김명진,윤필영,신동준,김수경,김종원,김규식 대한악안면성형재건외과학회 2000 Maxillofacial Plastic Reconstructive Surgery Vol.22 No.2

Since callus distraction technique was applied clinically for the correction of dentofacial deformity to the patients with hemifacial dysplasia by McCarthy in 1992, many surgeons have tried to apply this method to the maxillofacial region. But this technique has some drawbacks. One of the disadvantages of this technique is extensive scar formation in the facial area, which is a sequelae of extraoral approach for supraperiosteal dissection of the periosteum overlying the mandible. Recently, we have made an effort to perform this technique through intraoral approaches to prevent scar formation on the submandibular area and modified the design of the osteotomy, that is step osteotomy technique, to increase the raw bone surface on both osteotomized segments. The rationale for the application of this step osteotomy technique is to increase the amount of regeneracted bone and the length of distraction, to avoid damage of inferior alveolar neurovascular bundle, and to increase initial stability of the splitted segments. Step osteotomy procedure can be done with fine micro-osteotomy saw through subperiosteal tunneling. Extraoral pins should be inserted before making the osteotomy. Since 1974 we have applied this technique at 8 sites In 5 patients with mandibular deficiencies: 2 cases of hemifacial microsomia, 1 case of developmental facial asymmetry and 2 cases of mandibular bony defect. Mandibular elongation have been achieved from 12 to 20mm in length. 1 out of 8 site, we experienced non-union in the case of mandibular body defect. Some skeletal relapse and growth retardation phenomenon have been observed in some cases with the longest follow-up of 48 months.

냉동 제대혈 세포의 체외 증폭

김삼용,김철희,배광봉,김현수,박상준,김종숙,윤환중,조덕연 충남대학교 암연구소 1998 癌共同硏究所 硏究誌 Vol.2 No.1

Background : Cord blood(CB), which has no HLA restriction, is an alternative to bone marrow for hematopoietic stem cell transplantation. The use of cord blood, however, is limited by the number of progenitor/stem cells necessary to reconstitute the older child or adult. Therefore, ex vivo expansion of CB could have tremendous impact on diverse clinical settings. We studied the ex vivo expansion of isolated population of CD34_(+) cells from cryopreserved CB cells. Methods : CD34 cells were isolated from cryopreserved CB mononuclear cells. Purified cells were cultured with various combinations of hematopoietic growth factors including erythropoietin(EPO), stem cell factor(SCF), granulocyte-colony-stimulating factor(G-CSF), gra-nulocyte, macrophage-colony-stimulating factor(GM-CSF), interleukin-1β(IL-1β), 1L-3, and IL-6. After 7, 10 or 14 days of culture, the fold increases of colony-forming unit- granu-locyte, macrophage(CFU-GM), burst-forming unit-erythroid(BFU-E), colony-forming unit-mix (CFU-Mix), and high proliferative potential colony-forming cell(HPP-CFC) were evaluated. Results : Ten-day culture with the combination of EPO, SCF, G-CSF, IL-1β, and IL-3 resulted in a median of 60-fold increase of CFU-GM, which was greater than those with the combinations of less than 5 growth factors. The addition of IL-6 or GM-CSF to this combination did not enhance CFU-GM expansion. Ten-day culture was significantly superior to 7-day culture for CFU-GM expansion. Prolongation of culture to 14 days, however, revealed decreased expansion of CFU-GM compared to 10 days. BFU-E and CFU-Mix were expanded to 2~5 folds in 7-day culture with the combination of EPO, SCF, and G-CSF. Further expansion was not achieved in 10-day culture and colonies disappeared in 14-day culture. HPP-CFC was expanded to a median of 7.5 folds in 7-day culture with the combination of EPO, SCF, G-CSF, IL-1β, IL-3, and IL-6. Neither 10-day or 14 day-culture enhanced expansion of HPP-CFU. Conclusion : Cryopreserved cord blood cells maintain ex vivo expansion potential. In our system, 10-day culture with the combination consisting of EPO, SCF, G-CSF, IL-1β, and IL-3 seems to be adequate for hematopoietic progenitor/stem cell expansion from cryopreserved cord blood cells.

용혈성 요독 증후군이 합병된 출혈성 장염 1예

김능수,이원길,김기연,이종명,윤종수,김성한,전정훈 대한감염학회 1997 감염 Vol.29 No.3

저자들은 쇠고기에 의해 매개된 것으로 추정되는 출혈성 장염 및 이에 합병된 용혈성 요독 증후군 환자 1예를 경험하였기에 문헌고찰과 함께 보고하는 바이다. We report case of hemolytic uremic syndrome(HUS) associated with hemorrhagic colitis. A 35-year0old man was admitted to a local hospital because of abdominal pain and watery diarrhea which had developed about 6 hours after taking roast beef. He was treated with intravenous fluids and antibiotics, but watery diarrhea changed to bloody in nature from the next day. He was transferred to our hospital due to progressive ascites and jaundice on his 8th day of illness. Examinations revealed ascites, jaundice, microangiopathic hemolytic anemia, thrombocytopenia and uremia. Sorbitol-negative Escherichia coli was isolated from his stool, which proved as Shiga-like toxin-negative E. coli, serotype O25. His conditions improved markedly after three times of plasmapheresis and intravenous fluids therapy, and the organism was not isolated from the follow-up stool culture.

항암제 처리한 백혈병 세포주에서의 Apoptosis 발현

김삼용,윤소현,김현수,김종숙,윤환중,김진경,조덕연 충남대학교 암연구소 1998 癌共同硏究所 硏究誌 Vol.2 No.1

The bcl-2 proto-oncogene encodes a 26 kD protein that promotes cell survival by blocking apoptosis. The bax protein is a member of the bcl-2 familly, now known to form heterodimers with the bcl-2 protein. The ratio of bax to bcl-2 is be critical in determining the fate of the cell in response to stimuli that can induce apoptosis. Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro with Topo I inhibitory action. In the present study, the relationship between apoptosis and the apoptosis related proteins, bcl-2 and bax were investigated in human leukemic cell lines HL-60, U-937 and CEM-CM3. All anticancer drugs(adriamycin, etoposide, camptothecin, SB-31) induced extensive apoptosis in HL-60, U-937 cells and CEM-CM3 cells. The expression of bcl-2 and bax protein were determined in cell lines by western blotting before and after incubation with anticancer drugs at different time points. 1) In HL-60 or U-937 cell lines, down regulation of bcl-2 and up-regulation of bax were found after incubation with ADR, VP-16 or camptothecin. 2) In HL-60 or U-937 cell lines, no significant change in bcl-2 or bax protein expression resulted after incubation with SB-31. 3) In CEM-CM3 cells, virtually no change was noted in bcl-2/bax expression after incubation with ADR, VP-16, camptothecin or SB-31. It is suggested that different leukemic cell lines use different pathways of apoptosis activation and a given cell may utilize different pathways of apoptosis activation in response to different cytotoxic agents.

Breeding of Tetraploid in Codonopsis lanceolata (Sieb. et Zucc.) Trautvetter by Colchicine Treatment

Kim,Ik-Hwan,Kim,Hag-Hyun,Hong,Eui-Yon,Yun,Jong-Sun,Yun,Tae,Hwang,Ju-Kwang,Lee,Cheol-Hee 한국자원식물학회 2003 Plant Resources Vol.6 No.3

Present studies were carried out to produce tetraploid plants by colchicine treatment using seeds, seedlings and shoot tips of Codonopsis lanceolata. Three tetraploid plants of C. lanceolata were produced from seeds which absorbed 0.1 % colchicine solution for 12 hours, and 0.5% colchicine solution for 1 and 6 hours from seedlings, respectively. But tetraploid was not produced from shoot tips treated by colchicine solution. Compared to diploid, tetraploid plants had larger stomata, but less number of stomata. Fresh weight of tetraploid plants was 1.4∼3.6 times heavier than diploid plants.