고등급 장액성 난소암의 예후에 영향을 미치는 저산소증 연관 유전자 발현 연구

학위논문 (박사)-- 서울대학교 대학원 의과대학 의학과, 2017. 8. 박노현.Background: Ovarian cancer is the most lethal disease among malignancies of the female genital tract because of a higher rate of diagnosis at advanced-stage disease. Thus, maximal debulking surgery followed by adjuvant chemotherapy is essential to treat ovarian cancer. However, drug resistance of ovarian cancer is expected to be associated with microenvironment, which may hinder the improvement of prognosis and the development of optimal biomarkers for ovarian cancer. Among various types of tumor microenvironment, hypoxia-related genes are known to be associated with carcinogenesis, proliferation and invasion of tumor cells. However, they have been not investigated sufficiently for their roles affecting drug resistance and prognosis of ovarian cancer. Thus, we evaluated expressions of hypoxia-related genes in the most common histologic type, high-grade serous adenocarcinoma of the ovary (HGSO), and investigated the roles of hypoxia-related genes affecting platinum-resistance and survival of HGSO. Methods: We determined IC50 of cisplatin, and then evaluated cell viability after treatment of cisplatin under hypoxia in OV90 cells. Basic mRNA levels of CBP, P300, HIF-1α, HIF-1β, FIH and VHL were evaluated in each five tissues of normal ovary and HGSO by using of quantitative RT-PCR. Then, we investigated protein levels after treatment of cisplatin under hypoxic conditions (normoxia, 5%O2, 3%O2 and 1%O2) in OV90 cells by using Western blot. Furthermore, we estimated cellular proliferation and reactive oxygen species (ROS) production in OV90 cells by N-acetylcysteine (NAC) to determine relationship between hypoxia and ROS generation. Finally, we performed immunohistochemistry (IHC) for the six genes in patients with HGSO (n=149), and investigated prognostic factors affecting platinum-resistance and progression-free survival (PFS) among clinico-pathologic factors and the six hypoxia-related genes. Results: IC50 of cisplatin were 50 μmol/L in OV90 cells, and hypoxic cells were less inhibited than normoxic cells after treatment of IC50 of cisplatin. In HGSO, mRNA levels of CBP, P300, HIF-1α and HIF-1β were lower than those in normal tissues, whereas mRNA levels of FIH and VHL were higher than those in normal tissues. In Western blot, the protein level of HIF-1β was increased, whereas that of VHL was decreased under hypoxic conditions after treatment of cisplatin in OV90 cells. Furthermore, the protein level of HIF-1β was increased, whereas that of VHL was decreased when the concentration of NAC was increased with cisplatin in OV90 cells. On IHC, a low expression of HIF-1β and a high expression of VHL were favorable factors for reducing platinum-resistance in patients with HGSO (Adjusted ORs, 0.657 and 0.38195% CI, 0.443-0.975 and 0.165-0.878). In addition, a low expression of HIF-1β and a high expression of VHL were related with improved PFS in patients with HGSO (adjusted HRs, 0.657 and 0.38195% CIs, 0.443-0.975 and 0.165-0.878). Conclusions: A high expression of HIF-1 β and a low expression of VHL may be related with platinum-resistance and poor prognosis in patients with HGSO.Introduction 1 Materials and methods 3 Results 9 Discussion 21 References 26 Abstract (Korean) 31 Supplements 33Docto

(A) study on the phytate content and protein digestibility of various soy products

본 연구는 여러 종류의 두류 및 대두 식품인 비지, 두유, 두부, 청국장, 메주, 간장, 된장을 실험실에서 제조하거나 구입한 다음 가공 또는 발효에 따른 phytate 함량의 변화를 분석, 비교하였다. 또한 천연적으로 존재하는 phytate 함량이 단백질 소화에 미치는 영향을 알아보기 위하여 각 대두 식품들의 pepsin, pancreatin 소화율을 측정하였으며 phytate 함량과 단백질 소화율 간의 상관관계를 구하였다. 이와 아울러 각 식품들의 수용성 단백질 분획의 변화 양상을 알기 위하여 전기 영동을 실시하였다. 본 실험에서 얻어진 결과를 요약하면 다음과 같다. 1. 콩의 phytate 함량은 2.4%이었고, 가공에 따라 두유는 크게 줄어 0.1-0.2%, 두부는 0.1-0.3%, 비지는 약 0.2%이었다. 발효에 의해 청국장은 0.35%, 메주는 0.6%, 간장은 0.2%, 된장은 0.1-0.4%로 줄었고 발아된 콩나물도 0.06%로 감소하였다. 2. 가용성 단백질 함량은 콩과 제조 두유에서 가장 높았고 다음은 구입 두유, 발효 식품의 순이었으며 비지와 두부는 비슷한 수준으로 낮았다. 3. Pepsin에 의한 대두 식품의 소화율은 시간이 지남에 따라 증가하였고 제조 두유를 제외하고는 콩이 가장 높았다. 발효 식품들은 24시간 후의 소화율이 낮은 경향이었다. phytate/protein 비율과 pepsin에 의만 소화율(30분)간에는 상관관계가 없는 것으로 나타났다. 4. Pancreatin에 의한 소화의 경우 pepsin에 의한 소화에서와는 달리 비지, 두유, 두부등 콩을 가공하였을 때 소화율이 향상되었고 발효 식품들의 소화율도 높게 나타났다. phytate/protein 비율과 pancreatin 소화율(4시간)은 유의적인 부(負)의 상관관계를 나타내었다 (p<0.01, r=-0.58). 5. 대두 식품의 수용성 단백질 분획을 전기영동법을 통하여 살펴본 결과 콩을 불렸을 때 실온에서 잘 녹는 수용성 성분의 bands가 사라지는 변화를 보였다. 비지와 두부는 유사한 단백질을 가졌으나 비지에는 저분자량의 bands가 많았다. 발효 식품들은 전기영동상 감지할 수 없을 정도의 저분자량으로 분해되어 나타났다. 이상의 결과로 미루어 보아 대두 식품의 발효 및 가공중 단백질의 많은 변화가 있을 것으로 생각되며 발효 식품의 경우 그 변화가 뚜렷한 것을 알 수 있었다. 대두 식품 중 phytate가 pepsin 소화에는 큰 영향을 미치지 못했으나 pancreatin 소화에는 많은 영향을 주었는데 이 기전에 대한 연구가 앞으로 이루어져야 할 것이다.;This study was undertaken to find out the effect of phytate which naturally exists in various soybean foods on the protein digestibility. The change in the phytate content due to the physical and fermentation processes of curd residue, soy milk, bean curd, cheongkukjang, meju, soy sauce, and soy paste made with soybean was analyzed and compared with purchased soy products. Protein digestibility was tested by pepsin and pancreatin for the various soy foods and correlation between phytate content and protein digestibility was examined. The results revealed from this study can be summarized as follows: 1. The phytate content of soybean was 2.4%, and this content decreased as it was processed. Thus its content in soy milk, bean curd, and curd residue was 0.1-0.2%, 0.1-0.3%, and 2%, respectively. The phytate content of fermented cheongkukjang, meju, soy sauce, soy paste and germinated bean sprouts also decreased from 2.4% in raw soybean to 0.35%, 0.6%, 0.2%, 0.1-0.4% and 0.06%, respectively. 2. Soybean and lab-made soy milk sample showed the highest level in soluble protein content: the next was purchased-soy milk, and fermented soy products. Curd residue and bean curd were the lowest in the content. 3. Protein digestibility created by pepsin increased as time passed. The digestibility of soybean was the highest among tested products except lab-made soy milk: the digestibility of fermented soy products was at a low level after 24 hours compared to other soy products. Correlation between phytate/protein ratio and protein digestibility by pepsin (after 30 min) was insignificant. 4. The digestibility by pancreatin was enhanced when the soybean was processed to curd residue, soy milk, bean curd. In addition, the digestibility of fermented soy products was also high level. Correlation between phytate/protein ratio and pancreatin digstibility (after 4 hours) was significant (p<0.01, r=-0.58). 5. The results of SDS-PAGE for the soluble protein fractions of soy products are as follows: soaked bean showed the change in its bands pattern, i.e., the soluble components disappeared. Curd residue and bean curd had similar proteins but curd residue had many low molecular weight bands. Fermented soy products were degraded and they were not detected by SDS-PAGE. Judging from this result, it can be concluded that many changes of protein happened in the process of fermentation and manufacturing of soy products, and the change was especially clear in the case of fermented foods. Endogenous phytate contained in soy products didn't show any significant influence on the protein digestibility by pepsin, but showed a significant influence in pancreatin digestion. Therefore, a further research to elucidate this mechanism should be done in the future.목차 = ⅰ 논문개요 = ⅵ Ⅰ. 서론 = 1 Ⅱ. 실험 재료 및 방법 = 3 A. 시약 = 3 B. 식품시료 = 3 1. 콩류 = 3 2. 두유류 = 4 3. 두부류 = 4 4. 대두 발효 식품류 = 4 5. 탈지 대두분과 분리 대두단백 = 5 6. 기타 대두 식품 = 6 C. 식품 성분의 분석법 = 6 1. 수분 함량 및 단백질 함량 = 6 2. Total phytate 함량 = 6 3. 가용성 단백질 함량 = 7 D. 단백질의 소화율 측정법 = 9 1. Pepsin에 의한 소화 = 9 2. Pancreatin에 의한 소화율 = 11 E. Folin-Lowry법과 Macro-Kjeldahl법의 비교 = 11 F. 전기영동법 = 11 1. Stock solution = 12 2. Gel mixture와 gel의 제조 = 14 3. 전기영동의 실시 = 15 4. 염색과 흡광도 측정 = 16 5. 표준 단백질 곡선 = 17 Ⅲ. 결과 및 고찰 = 18 A. 제조 시료의 무게 = 18 B. 대두 제품의 수분, 단백질 및 Phytate 함량 = 19 C. 가용성 단백질 함량 = 23 D. 단백질의 소화율 = 25 1. Pepsin에 의한 소화 = 25 2. Pancreatin에 의한 소화 = 29 3. Pepsin 소화와 Pancreatin 소화와의 비교 = 33 E. Folin-Lowry법과 Macro-Kjeldahl법의 비교 = 34 F. 전기영동에 의한 대두 식품의 수용성 분획 비교 = 37 1. 실험실에서 제조한 대두 제품의 전기영동 = 37 2. 구입한 대두 제품의 전기영동 = 40 3. 발효 식품들의 전기영동 = 42 Ⅳ . 요약 및 결론 = 43 참고문헌 = 45 ABSTRACT = 5

Educational Need Assessment of Advanced Practice Nurses on the Pathophysiology

Purpose: Knowledge on pathophysiology is important to understand diseases and patient conditions and also in providing advanced nursing care. This study was performed to investigate the present situation, knowledge, and educational need on pathophyiology of advanced practice nurses (APN). Method: Eighty-four APN, who is practicing in hospitals nationwide, were surveyed with structured questionnaires developed by researchers. The questionnaire were composed of 109 items with 4 point scales with the contents of general characteristics, educational experience on pathophyisology, educational need, and knowledge on major and subcategories of pathophysiology. Result: The 77.1% of APN had experience of taking pathophysiology lecture and the lecturer were mainly medical doctors (39.8%) and APN (32.4%). The 61.5% of APN answered that the knowledge on pathophysiology was helpful and 97.6% of APN thought pathphysiologic education is needed in clinical practice. Seven major categories of pathophysiology showed higher scores were water imbalance, heart diseases, hemodynamic disorders, arterial vessel disorders, electrolyte imbalance, diseases of the digestive system, and diseases of the respiratory systems. Educational needs on the major and subcategories of pathphysiology were different by the field of APN and also by the clinical experiences as APN. APN also responded that there is a necessity to change the present curricular more specific to each practice field and more focus on the clinical practice. Conclusion: It was confirmed that educational and clinical need' on the pathophysiology is very high and there is urgent need to change current curricular more specific to the field of APN and also clinical practice. This survey will help to identify and clarify the areas of change and to improve the education for the APN.본 연구는 2006년도 대한기초간호자연과학회 정책연구비 지원으로 수행되었음

Surgical management of recurrent ovarian cancer

Most patients with advanced-stage epithelial ovarian cancer will experience a relapse of disease despite a complete response after surgical cytoreduction and platinum-based chemotherapy. Treatment of recurrent ovarian cancer mainly comprises various combinations of systemic chemotherapy with or without targeted agents. The role of cytoreductive surgery for recurrent ovarian cancer is not well established. Although the literature on survival benefit of cytoreductive surgery for recurrent disease has expanded steadily over the past decade, most studies were retrospective, single-institution series with small numbers of patients. Given the balance between survival benefit and surgery-related morbidity during maximum cytoreductive surgical effort, it is essential to establish the optimal selection criteria for identifying appropriate candidates who will benefit from surgery without worsening quality of life. Three phase III randomized trials for this issue are currently underway. Herein, we present contemporary evidence supporting the positive role of cytoreductive surgery and offer selection criteria for optimal candidates for surgery in the treatment of recurrent ovarian cancer. (C) 2016 Elsevier Inc. All rights reserved.OAIID:RECH_ACHV_DSTSH_NO:T201604232RECH_ACHV_FG:RR00200001ADJUST_YN:EMP_ID:A080402CITE_RATE:4.959DEPT_NM:의학과EMAIL:[email protected]_YN:YN

An umbrella study of biomarker-driven targeted therapy in patients with platinum-resistant recurrent ovarian cancer: a Korean Gynecologic Oncology Group study (KGOG 3045), AMBITION

A pilot study of biomarker-driven targeted therapy in patients with platinum-resistant recurrent ovarian cancer has been started in Korea. Archival tumor samples were tested for HRD and PD-L1 status. Treatment arms will be allocated according to the test results. For HRD+ patients, we tested the synergistic effects of olaparib and other agents; treatment arms will randomly be allocated. (Arm 1: olaparib and cediranib; Arm 2: olaparib and durvalumab). For HRD- patients, we tested the role of biomarker-driven immunotherapy according to PD-L1 expression (Arm 3: durvalumab and chemotherapy in patients with high PD-L1 expression; Arm 4: durvalumab, tremelimumab, and chemotherapy in patients with low PD-L1 expression). Sixty-eight patients will be included from three Korean institutions within 1 year. The primary endpoint is the response rate according to RECIST 1.1 (6 months after treatment initiation).OAIID:RECH_ACHV_DSTSH_NO:T201916095RECH_ACHV_FG:RR00200001ADJUST_YN:EMP_ID:A080402CITE_RATE:2.183DEPT_NM:의학과EMAIL:[email protected]_YN:YY