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Zebrafish 동물모델에서 human HtrA2의 expression system 정립에 관한 연구
조성원,박효진,김구영,남민경,김호영,고인호,김철희,임향숙,Cho, Sung-Won,Park, Hyo-Jin,Kim, Goo-Young,Nam, Min-Kyung,Kim, Ho-Young,Ko, In-Ho,Kim, Cheol-Hee,Rhim, Hyang-Shuk 한국생명과학회 2006 생명과학회지 Vol.16 No.4
Mitochondrial serine protease로 알려진 human HtrA2 (hHtrA2)는 apoptosis 유도 과정에서 중요한 역할을 담당하고 있을 뿐만 아니라 hHtrA2가 motor neuron degeneration과 관련이 있다는 최근 연구 결과가 있으나, hHtrA2의 생리적 기능은 아직 명확하게 밝혀져 있지 않다. 이와 같이 생체내에서 필수적인 업무를 담당하는 hHtrA2의 기능을 심도 있게 연구하기 위해서는 적절한 동물모델 시스템이 필요하나 이에 대한 연구도 미흡한 실정이다. 따라서 본 연구에서는 hHtrA2의 기능 분석을 위한 기본적인 실험으로 zebrafish라는 동물모델을 선택하여 hHtrA2의 발현 시스템을 정립하였다. 먼저 zebrafish에 hHtrA2를 발현시키기 위하여 zebrafish에서 일반적으로 사용되는 발현 시스템인 pCS2+ vector에 hHtrA2와 GFP를 cloning하고 plasmid를 HEK293 cell에 transfection한 후, hHtrA2-GFP fusion 단백질의 발현을 immunoblot과 immunofluorescence staining assay로 확인한 바 약 64 kDa의 hHtrA2 단백질의 발현을 확인할 수 있었다. Zebrafish에서 hHtrA2-GFP fusion 단백질의 발현양상은 immunofluorescence microscope으로 확인하였다. hHtrA2-GFP DNA와 mRNA를 zebrafish embyro에 microinjection하여 두 가지 component의 발현을 비교 분석한 결과, DNA는 dot 형태로 mRNA는 몸 전체에 퍼져보이는 형태로 발현 양상의 차이는 있었으나 둘 다 zebrafish embryo에서 잘 발현되는 것을 알 수 있다. 다음 DNA를 주 component로 microinjection하여 zebrafish embryo에서 발현을 확인한 결과 hHtrA2는 72 hpf 까지 발현이 지속되는 것을 확인하였다. 본 연구에서 정립한 hHtrA2의 zebrafish 발현 조건은 앞으로 zebrafish에서 hHtrA2의 생리적 기능을 심도있고 정확하게 연구하는 데 있어 기본적인 자료로 활용 할 수 있을 것이다. HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.
400 kHz 페라이트 유도 결합 플라즈마를 위한 임피던스 매칭 네트워크
조성원,방진영,이영광,정진욱,Cho, Sung-Won,Bang, Jin-Young,Lee, Young-Kwang,Chung, Chin-Wook 한국반도체디스플레이기술학회 2007 반도체디스플레이기술학회지 Vol.6 No.2
Low frequency (400 kHz) Ferrite ICP has been proposed for large area processing. Because the coupling coefficient is close to 1, the transformer matching can be adapted to Ferrite ICP. The transformer matching system is simple. In this paper the new matching system by controlling the turns of transformer using relays is proposed for impedance matching. We developed a simple matching system and characterized it when the turns were changed. It was observed that the 2-channel relay is available for transformer matching.
위장관 ( 胃腸管 ) : 위암환자에서 내시경적 초음파단층법 ( EUS ) 에 의한 TNM분류의 평가
조성원(Sung Won Cho),심찬섭(Chan Sup Shim),김진홍(Jin Hong Kim),이문성(Moon Sung Lee),조영덕(Young Deok Cho),손진휘(Jin Hwe Son),김홍수(Hong Su Kim) 대한소화기학회 1991 대한소화기학회지 Vol.23 No.3
N/A EUS-a combination of endoscopy and ultrasonography was developed to improve the diagnostic accuracy of ultrasound by direct image of the target lesion via the gastrointestinal lumen. This new technique will prove to be accurate in the staging of gastric malignant tumors because of the clear visualization of the tumor infiltration and of involvement of adjacent lymph nodes. EUS was performed preoperatively for assessing the clinical TNM classification in 54 patients with a gastric carcinoma confirmed by endoscopic biopsy. The results were correlated with those obtained with the histology of resected specimens according to the pathological TNM classification. 1) In 54 patients with a gastric carcinoma, the mean age of the patients was 55.9 years with a male to female ratio of 1.84 : l 2) Early gastric cancer was diagnosed with the histology of resected specimens in 7 patients, and advanced gastric cancer in 47 patients. In 4 cases of advanced gastric cancer, only exploratory laparotomy was performed, so that the extent of tumor infiltration could not be assessed histologically. 3) In assessing the depth of tumor invasion, the stage accuracy rate of EUS was 85.6%. for T1 tumors, 83.3% for T2 tumors, 76.5% for T3 tumors, and 85.7% for T4 tumors. The overall accuracy rate was 82.0% (41/50). 4) In staging lymph nodes, the accuracy rate of EUS was 57.9% for NO, 75.0%. for Nl, and 54.5% for N2. The overall accuracy rate was 64.8% (35/54). A rate of false-positive diagnosis was 42.1% and a rate of false-negative diagnosis was 14.3%. 5) In assessing distant metastases, the accuracy rate of EUS was 100% for MO, 57.1% for Ml. In 2 patients with metastases in the right lobe of the liver and 1 patient with metastasis in the ovary, metastases were not visualized with EUS due to its limited depth of the penetration of ultrasound. 6) In stage grouping of stomach cancer, the overall accuracy rate of EUS was 57.4% (31/54). In conclusion, EUS will become an important diagnostic procedure for clinical TNM classification with the combined use of transcutaneous abdominal ultrasonography or abdomimal computed tomo-graphy.
비면역기능저하 환자에서 발생한 비안와 뮤코르진균에 의한 단안 압박성 시신경병증
조성원(Sung-Won Cho),하석규(Suk Gyu Ha),김승현(Seung-Hyun Kim) 대한검안학회 2019 Annals of optometry and contact lens Vol.18 No.1
Purpose: To report a case of unilateral compressive optic neuropathy caused by rhino-orbital mucormycosis in an immunocompetent patient. Case summary: A 62-year-old male visited the ophthalmology department with discomfort and decreased visual acuity in his right eye. There were no signs of eyelid edema, limited ocular movement, or proptosis. The patient had no history of diabetes and hypertension. On ophthalmologic examination, relative afferent pupillary defect, decreased color vision, optic disc swelling, and inferior visual field defect were observed in the right eye. There were no abnormal results of laboratory investigation. Brain and orbit magnetic resonance image revealed right ethmoidal sinus mass-like lesion with extension into the orbital cavity, and ethmoidal biopsy confirmed it as mucormycosis. Conclusions: We reported a case of unilateral compressive optic neuropathy caused by rhino-orbital mucormycosis in the non-immunocompromised patient without other systemic diseases.