Characteristics and clinical significance of circulating tumor DNA in extranodal NK/T cell lymphoma

김진주 Graduate School, Yonsei University 2022 국내박사

Background: Extranodal Natural killer/T-cell lymphoma (ENKTL) is an aggressive subtype of non-Hodgkin lymphoma. Necrosis is a classic morphologic feature of this disease which makes it difficult to obtain a satisfactory tissue biopsy for analysis. Somatic gene mutations identified in ENKTL tissue provide information about the pathogenesis and prognosis of ENKTL. ctDNA provides real-time mutational information that can be used to detect and monitor biomarkers of cancer. The recent introduction of a highly sensitive next generation sequencing (NGS) approach allows analysis of circulating cell-free DNA (ctDNA) using plasma. We conducted a study to assess the utility of ctDNA genetic mutation analysis on ENKTL. Methods: The analysis was performed on samples collected from 34 newly diagnosed and 11 relapsed patients. Longitudinal samples were collected from patients undergoing asparaginase (ASP)-based chemotherapy and from relapsing patients undergoing treatment with chemotherapy and immunotherapy. ctDNA was investigated in association with clinicopathological features of ENKTL through the Ann Arbor stages of ENKTL, Epstein-Barr virus DNA quantification, treatment response, and treatment modification. The concordance of somatic variants between plasma and matched tissues was also evaluated. Results: Tumor tissue biopsy sequencing was used to evaluate the performance of ctDNA. ctDNA had an overall sensitivity of 77.8% in identifying tissue-confirmed pathogenic somatic mutations. ctDNA concentration was correlated with treatment response, stage, and EBV DNA quantitation. BCOR was the most frequently mutated gene in newly diagnosed patients, and so was shown to be a suitable adverse prognostic marker, followed by TP53 and DDX3X mutations. Longitudinal ctDNA profiling identified treatment-dependent patterns of clonal evolution in patients who relapsed after chemotherapy. TP53 was the most frequently mutated gene in samples from relapsed/refractory patients, indicating a possible contribution of TP53 mutation to disease relapse. Conclusions: ctDNA analysis is a non-invasive tool which was shown to facilitate monitoring of tumor burden. It may be used to compliment EBV quantitation and imaging studies. Moreover, ctDNA may be used to predict chemo-refractory patients and may serve as a prognostic biomarker in patients with ENKTL.

Circulating tumor cells in breast cancer with orthotopic allograft mouse model

한애리 Graduate School, Yonsei University 2015 국내박사

Breast cancer causes death not because of the primary tumor in the breast but because of metastases in distant sites that gradually cause organ dysfunction.1 Circulating tumor cells (CTCs) are cells that have detached from the primary tumor or metastatic tumor sited and entered the peripheral circulation.2 We established orthotopic breast cancer animal model of breast cancer with CTCs and validated the role of surgical treatment in the subjects with primary breast cancer without CTCs. (Materials and Methods) We implanted 1 ?104 GFP expressing 4T1 cells in 4th or 5th mammary fat pad of 8 week-old BALB/cAnNCrl mice(n=69). For the evaluation of role of surgery in early breast cancer with no CTC, we removed the tumor cell just before CTCs appeared in circulation (n=35). After removal of the primary tumor we evaluated the extent of surgical procedures. If there was any suspected residual tumor tissue in operation bed, the procedure was categorized as R1 resection. If operation bed was not suspected with residual tumor tissue, the resection was categorized as R0 resection.(Results) All the experimental animal showed CTCs after primary tumor appeared (n=69). Furthermore, tumor volume and the number of CTCs increased over time and tumor volume showed statistically significant correlation with CTCs (p<0.001).Next, we removed primary tumor before a CTC was detectable with flow cytometry assay (n=35). The mice showed no CTCs during follow up if removal of the tumor results in complete resection of the tumor, R0 resection (n=25). However, we could detect CTCs during follow up period if the resection results in simple wide excision, R1 resection (n=10) (p<0.001).(Conclusions) We successfully developed orthotopic allograft mouse model with breast cancer and circulating tumor cells, which is most similar with human breast cancer evolution. We also showed R0 resection of the primary tumor could result in no CTCs during follow up period if the resection was done when there was no CTCs. We believe this finding could be translated in the clinical field.

(The) impact of circulating tumor DNA in biliary tract cancer under chemotherapy

윤우빈 Graduate School, Yonsei University 2023 국내박사

Biliary tract cancer (BTC) is a highly aggressive cancer with a very poor prognosis. In general, the incidence of BTC is higher in Eastern countries than in Western countries. BTC was once considered a geographically region-specific disease. However, according to recent reports, the incidence of BTC has increased globally. Most patients with BTC were first diagnosed at the advanced stage because the disease is usually asymptomatic during the early stage. Tissue biopsy is the current gold standard for cancer diagnosis, but this invasive technique has challenges. Despite the increased incidence rate and poor prognosis of BTC, understanding this disease is still not satisfactory. To discover actionable target genes and monitor the drug response of patients, we enrolled unresectable BTC patients (n = 41), and circulating-tumor DNA (ctDNA) from plasma samples was collected at multiple timepoints while patients received chemotherapy (pre-1st chemotherapy, pre-2nd chemotherapy, pre-4th chemotherapy, and progression disease). All samples were deep sequenced with a large panel containing 531 pan-cancer genes. We identified highly observed variants, such as TP53, ARID2, KRAS, ARID1A, PDE4DIP, ARID1B, CHD4, FAT1, PIK3CA, SPEN, APC, ATM, ATR, ERBB4, FGFR2, and IDH1. In addition, copy number alterations (CNAs) of MYC, ERBB2, CDKN2A, GATA4, ARID2, MDM2, PIK3R3, CDK12, and EGFR were observed. Key pathways and genes were curated from the literature and detected single nucleotide variants (SNVs) were categorized by them. Epigenetic regulation, TP53 signaling, the PI3K/AKT/mTOR and RAS/RAF/ERK pathways, DNA damage, angiogenesis, and DNA repair were highly ranked. TP53, ARID2, and PTPRT frequently occurred under chemotherapy. In particular, the PTPRT mutation remarkably increased in a cohort with progression disease as compared with that of cohorts at other timepoints. The survival rate of BTC patients with a low tumor mutation burden (TMB) was higher than that of the high TMB patient group. Also, a new threshold by delta blood TMB (dTMB) showed potential as a marker for diagnosis. In the present study, we suggested the advantages of cell-free DNA (cfDNA)-targeted sequencing and discussed candidates of precision therapy and understanding molecular profiling of BTC patients under chemotherapy.

Characteristics of circulating-tumor DNA in non-metastatic clear cell renal cell carcinoma

김홍경 Graduate School, Yonsei University 2023 국내박사

Background: Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma. Circulating-tumor DNA (ctDNA) has emerged as a biomarker which complements or acts as an alternative to renal mass biopsy. However, the characteristics of ctDNA have not been elucidated in non-metastatic ccRCC, especially with small tumors. Methods: Plasma was preoperatively collected from 120 patients who underwent surgical resection for suspected kidney cancer. Samples of ccRCC were sequenced using next-generation sequencing and sequenced data were analyzed using the Pi-Seq algorithm (Dxome, Sungnam, Korea). The characteristics of ctDNA were compared between non-metastatic ccRCC and metastatic ccRCC. Non-metastatic ccRCC was stratified according to pathological T (pT) stage into pT1a and pT1b-3a, and associations with ctDNA were further investigated. The detection rate, variant allele frequency, and proportion of genes with ctDNA were evaluated. ctDNA was investigated in association with several clinicopathological features of ccRCC. The positive concordance of somatic variants between plasma and matched tissue was evaluated. Results: Of the 120 patients included in this study, 90 were diagnosed with ccRCC, 20 were diagnosed with non-ccRCC, and 10 were diagnosed with benign tumors. Among the 90 patients with ccRCC, 15 were excluded based on their medical history; and of the remaining 75 patients, non-metastatic ccRCC was confirmed in 67 patients. Most non-metastatic ccRCC cases (79.1%) were classified as pT1a ccRCC. Detection rates of ctDNA were 26.9% and 75.0% in non-metastatic and metastatic ccRCC, respectively. The detection rate of non-metastatic ccRCC showed a tendency to increase as the tumor size increased. The detection rate of ctDNA in pT1a ccRCC was 22.6%. Median variant allele frequencies of ctDNA were 0.351% and 1.168% in non-metastatic and metastatic ccRCC, respectively. The proportion of genes with ctDNA in non-metastatic ccRCC was different from that of metastatic ccRCC. VHL, PBRM1, SETD2, and BAP1 were frequently detected in metastatic ccRCC while NF1, TP53, and KDM6A were frequently detected in non-metastatic ccRCC. ctDNA detection in non-metastatic ccRCC was associated with tumor sizes and patient age, but not with tumor grade. ctDNA was frequently detected when lymphovascular invasion, fat tissue invasion, or venous tumor thrombus were concurrently observed, but the associations were not statistically significant. Positive concordance between ctDNA and matched tissue was poor in non-metastatic ccRCC. Conclusions: The characteristics of ctDNA in non-metastatic ccRCC were explored, with particular attention on small-sized pT1a ccRCC. Low detection rate, low variant allele frequency, and different proportion of genes with ctDNA were demonstrated in non-metastatic ccRCC compared with metastatic ccRCC. ctDNA was associated with tumor size and patient age in non-metastatic ccRCC. However, the relationship between ctDNA and tumor grade was not clear. Possible variants of clonal hematopoiesis were not filtered, which was a limitation of this study. A prospective study is required to demonstrate the clinical significance of ctDNA in non-metastatic ccRCC. Improvement of the sensitivity of ctDNA analysis and filtration of clonal hematopoiesis may advance the clinical utility of ctDNA. 연구배경: 투명세포 신세포암 (clear cell renal cell carcinoma; ccRCC) 은 신세포암에서 가장 많이 발견되는 유형이다. 순환종양 DNA (circulating-tumor DNA; ctDNA) 는 신장 종괴 생검을 대체하거나 보완할 수 있는 바이오마커 중 하나로 대두되고 있다. 그러나 비전이성 ccRCC 에서, 특히 작은 사이즈의 암종에서의, ctDNA 의 특성은 아직 밝혀지지 않았다. 연구방법: 신장암이 의심되는 종괴에 대하여 외과적 절제를 받은 120명의 환자로부터 수술 전에 혈장을 수집하였다. 그 중 ccRCC 환자의 혈장으로부터 추출한 DNA 를 차세대 시퀀싱 (Next-generation sequencing) 기술을 사용하여 시퀀싱 하였으며 Pi-Seq 알고리즘 (Dxome, 성남, 대한민국) 을 사용하여 ctDNA 를 분석하였다. 비전이성 ccRCC 의 ctDNA 의 특성은 전이성 ccRCC 와 비교되었다. 비전이성 ccRCC 를 pT1a 군과 pT1b-3a 군으로 나누어 ctDNA 와 연관된 조사를 추가적으로 실시하였다. 검출률, 변이의 대립유전자 빈도 및 ctDNA 가 검출된 유전자의 빈도가 평가되었다. ctDNA 는 여러 임상병리학적 특징과 연관되어 조사되었다. 혈장과 매칭된 조직 사이의 체세포 변이의 양성 일치성을 평가하였다. 연구결과: 120명의 환자 중 90명은 ccRCC, 20명은 non-ccRCC 그리고 10명은 양성 종양 환자였다. ccRCC 환자 중 15명의 환자는 특정 과거력으로 인해 연구에서 제외되었다. 75명의 ccRCC 환자 중 67명이 비전이성 환자로 확인되었다. 비전이성 ccRCC 의 대부분은 병리학적 T 단계 1a로 분류되었다 (79.1%). ctDNA 의 검출률은 비전이성과 전이성 ccRCC 에서 각각 26.9%와 75.0% 였으며, 비전이성 ccRCC 에서는 종양의 크기가 커질수록 ctDNA 검출률이 증가하는 경향을 보였다. pT1a ccRCC 의 경우 ctDNA 검출률이 22.6% 였다. 변이 대립유전자 빈도의 중간값은 비전이성과 전이성 ccRCC 에서 각각 0.351% 와 1.168% 였다. 비전이성 ccRCC 에서 ctDNA 가 검출된 유전자의 빈도는 VHL, PBRM1, SETD2 그리고 BAP1 이 주로 관찰되는 전이성 ccRCC 와는 차이가 있었다. NF1, TP53 그리고 KDM6A 가 비전이성 ccRCC 에서 자주 관찰되었다. 비전이성 ccRCC 에서 ctDNA 검출은 종양의 크기와 환자의 나이와 연관되었으며 종양의 등급과는 관련이 없었다. ctDNA 는 림프혈관계 침범, 지방조직침범 그리고 정맥종양혈전이 발견되는 경우에서 더 자주 관찰되었으나 통계학적인 차이는 없었다. 비전이성 ccRCC 에서 ctDNA 와 매칭된 조직 간의 양성 일치성은 좋지 않았다. 결론: 비전이성 ccRCC, 특히 pT1a 에 해당하는 작은 크기의 ccRCC 에 대하여 ctDNA 의 특성을 탐색하였다. 낮은 검출률, 변이의 낮은 대립유전자 빈도 그리고 전이성 ccRCC 와 다른 ctDNA 유전자 비율이 나타났다. 비전이성 암에서 ctDNA 는 종양의 크기 및 환자 나이와 연관되었으나 종양의 등급과는 그 관계가 불명확하였다. 여과되지 않은 클론 조혈 가능성이 있는 변이는 분석에서 고려되지 않았으며, 이는 이 연구의 한계 중 하나로 생각된다. 비전이성 ccRCC 에서 ctDNA 의 임상적 유용성을 입증하기 위해 전향적 연구가 실시되어야 한다. 민감도 개선 및 클론 조혈 변이 배제는 ctDNA 분석의 임상적 유용성을 향상시킬 것으로 사료된다.

순환식 병류형 벼·잡곡 겸용 건조기 개발

김동선 전북대학교 일반대학원 2016 국내박사

This study has been conducted to develop the Circulating-type Concurrent-flow Dryer for Rice and Minor Cereals. Circulating-type Cross-flow Dryer was generally used to dry rough rice only in korea. Barley, bean, corn and rapeseed were dried by way of sundrying in korea. In the way of sundrying happened decreasing of germination, labor increase and quality loss of harmful insects and other reason. So, it is necessary to develop a multipurpose dryer for rice and minor cereals. The specific objectives were as follows; 1. To simulate the drying process for rice and minor cereals 2. To determine the optimum design and operating factor of multipurpose dryer 3. To determine the optimum operating conditions of multipurpose dryer with the performance test The result were summarized as follows; 1. The simulation model to predict the characteristics of multipurpose dryer was developed and was validated through the drying tests for rough rice, barley, corn, bean and rapeseed. 2. The simulation program of multipurpose dryer was developed using MATLAB 7.3.0 (R2006b version). The basic structure of simulation program was consisted of main and sub program of thirteen. 3. Comparison of result between simulated and measured showed 0.97 R squared value and under about 10% difference. 4. Basic design factor of multipurpose dryer was determined using Box’s Complex algorithm. The optimum height of drying chamber was 0.5 m for the multipurpose dryer. The optimum volume ratio of drying chamber to tempering chamber was 1:4-6. 5. The performance of multipurpose dryer generally showed that the drying rate was 30-80% higher and energy consumption was about 10% lower than the other commercial dryer. 6. This dryer is possible to use in the wet climate country of east-southern asia. It is obvious that the developed multipurpose dryer is a good dryer because it has more efficient energy consumption and more rapid drying performance, and keeps the grain quality better simultaneously.

Clinical considerations in the use of forced-air warming blankets during orthognathic surgery to avoid postanesthetic shivering

박지연 서울대학교 대학원 2016 국내석사

Background: During head and neck surgery including orthognathic surgery, mild intraoperative hypothermia occurs frequently. Hypothermia is associated with postanesthetic shivering, which may increase the risk of other postoperative complications. To improve intraoperative thermoregulation, devices such as forced-air warming blankets can be applied. This study aimed to evaluate the effect of supplemental forced-air warming blankets in preventing postanesthetic shivering. Methods: This retrospective study included 113 patients who underwent orthognathic surgery between March and September 2015. According to the active warming method utilized during surgery, patients were divided into two groups: Group W (n = 55), circulating-water mattress; and Group F (n = 58), circulating-water mattress and forced-air warming blanket. Surgical notes and anesthesia and recovery room records were evaluated. Results: Initial axillary temperatures did not significantly differ between groups (Group W = 35.9 ± 0.7°C, Group F = 35.8 ± 0.6°C). However, at the end of surgery, the temperatures in Group W were significantly lower than those in Group F (35.2 ± 0.5°C and 36.2 ± 0.5°C, respectively, P = 0.04). The average body temperatures in Groups W and F were, respectively, 35.9 ± 0.5°C and 36.2± 0.5°C (P = 0.0001). In Group W, 24 patients (43.6%) experienced postanesthetic shivering, while in Group F, only 12 (20.7%) patients required treatment for postanesthetic shivering (P = 0.009, odds ratio = 0.333, 95% confidence interval: 0.147–0.772). Conclusions: Additional use of forced-air warming blankets in orthognathic surgery was superior in maintaining normothermia and reduced the incidence of postanesthetic shivering. 목 적: 양악 수술 또는 시간이 긴 악안면 종양 절제 및 재건 수술의 경우, 체온감소가 되는 경우가 빈번하고 회복실에서 환자는 오한을 느끼고 심한 전율이 발생하고 합병증의 위험성이 증가한다. 수술 시 체온관리 방법으로 수술침대에 가온패드를 사용하는 방법이 일반적이다. 하지만 가온패드 만으로 체온이 유지가 안되는 경우, 열효율이 좋은 에어워머의 사용이 적용될 수 있다. 양악수술을 받는 환자의 하체 부위에 에어워머를 사용하여 수술 중 체온을 유지하는 경우, 수술 후 오한과 전율을 감소시킬 수 있는지 알아보고자 하였다. 방 법: 2015년 3월 1일부터 2015년 9월 30일 까지 양악수술을 받은 모든 환자(113명)에서 질향상 과제로 에어워머를 구입하여 추가적으로 체온관리를 시행한 데이터가 있어 후향적으로 수술장부, 마취기록지, 회복실 기록지를 이용하여 가온패드만 적용되었던 환자군과 에어워머가 함께 적용되었던 환자군으로 나누어서, 액와 체온의 변화를 분석하고, 회복실에서 오한과 전율의 발생 빈도를 조사하였다. 결 과: 에어워머가 적용되지 않은 군(A군)이 55명, 에어워머가 적용된 군(B군)이 58명이었다. 두 군에서 성비, 나이, 체중, 수술시간에서 차이는 없었으며, 수술시작시 체온이 A군은 35.9±0.7℃이었고 B군은 35.8±0.6으로 차이가 없었으나. 수술종료 시 A군은 35.2±0.5℃이었고 B군은 36.6±0.5 (P = 0.04)로 차이가 있었다. 평균체온은 A군은 35.9±0.5℃이었고 B군은 36.2±0.5 (P = 0.0001)이었다. 또한 수술 후에 오한과 전율의 발생 빈도가 A군에서는 24명/55명 (43.6%) 였고, B군에서는 12명/58명(20.7%) 이었다(P = 0.009, odds ratio = 0.333, 95% CI: 0.147-0.772). 결 론: 본 결과로 양악수술 환자에서 에어워머를 적용하므로 수술실 내에서 환자의 체온이 저하되는 것을 방지 할 수 있었고 수술 후 회복실에서 오한과 전율의 발생 빈도를 낮출 수 있었다.

Identification of circulating transcripts associated with adenoma-carcinoma sequence in colorectal cancer

황다솜 Graduate School, Yonsei University 2022 국내박사

Colorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer-related mortality. The survival rate of CRC sharply declines with late detection of the cancer, so early diagnosis of CRC through regular screening is important. It is known that most CRCs develop through the adenoma-carcinoma sequence (ACS), which describes the transformation of normal epithelium to a non-advanced adenoma (NA) to an advanced adenoma (AA) and ultimately to a carcinoma. AA is known to have a high risk for developing CRC. Therefore, it is important to detect AA as well as CRC for decreasing the incidence of CRC. Currently, colonoscopy is available for screening AA and CRC, but it shows low compliance due to the invasiveness and inconvenience. Therefore, this study aimed to identify circulating transcripts that can be useful to develop a molecular assay for screening AA and CRC. For this purpose, the study was set up to identify circulating transcripts associated with the ACS. In brief, a total of blood samples was collected from subjects scheduled for colonoscopy, and the subjects were divided into 4 groups based on the results of the colonoscopy: (I) CRC, (II) AA, (III) NA, and (IV) control group. And then, the relative expression levels of circulating transcripts in the CRC, AA, NA, and control group were determined using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). The results showed that a total of 20 circulating transcripts were differentially expressed in each step of the ACS. Subsequently, expression profiles of the 20 circulating transcripts were analyzed by an artificial intelligence (AI) model to differentiate the AA and CRC group from the control and NA group. As a result, the sensitivity to detect AA and CRC was 75% and 89%, respectively, and the specificity was 72%. In conclusion, it seems that the 20 multiple circulating transcripts associated with the ACS may be useful to differentiate the AA and CRC group. 대장암은 전 세계에서 세 번째로 많이 발병하는 암이며 암과 관련된 사망에 있어 두 번째로 주요한 원인이 된다. 대장암은 진단되는 시기가 늦어 짐에 따라 생존율이 급격히 감소하므로 정기적인 검진을 통한 조기 진단이 중요하다. 또한 대부분의 대장암은 정상 상피세포에서 비진행선종, 진행선종을 거쳐 암으로 진행되는 선종-암화 기전을 통해 발생하는 것으로 알려져 있다. 진행선종은 선종 중에서도 암으로 발전할 확률이 높은 선종이므로 대장암의 발병율을 낮추기 위해서는 선별검사를 통해 이러한 진행선종을 검출하는 것이 중요하다. 그러나 기존에 주로 이용되는 검사법인 대장내시경은 침습적이고 불편하다는 단점으로 인하여 대상자들의 순응도가 낮다. 따라서 본 연구에서는 대장암 선별 검사법에서 유용하게 이용될 수 있는 선종-암화 기전과 연관된 혈중전사체들을 규명하고자 하였다. 먼저 대장내시경이 예정된 대상자의 혈액을 채취하고, 대장내시경 결과에 따라 대장암군, 진행선종군, 비진행선종군, 대조군으로 나누었다. 다음으로, 실시간 정량적 중합효소 증폭법(RT-qPCR)을 이용하여 대장암군, 진행선종군, 비진행선종군 및 대조군에서 혈중전사체들의 상대발현양을 분석하였다. 그 결과에 따르면 총 20개의 혈중전사체가 선종-암화 기전 단계에 따라 발현 양상에 유의한 차이를 나타냈다. 그 후 해당 20개 혈중전사체의 발현 프로파일을 기반으로 대장암군 과 진행선종군을 대조군 과 비진행선종군으로부터 구별하는 인공지능(AI) 모델을 구축하였다. 구축된 인공지능 모델의 성능을 확인한 결과에 따르면 대장암군과 진행선종군에 대한 민감도는 각각 89%와 75%였고, 비진행선종과 대조군에 대한 특이도는 72%로 나타났다. 본 연구의 결과는 대장암의 선종-암화 기전과 관련된 특정 혈중전사체들이 대장암군 및 진행선종군을 구별하는데 유용할 수 있음을 보여주고 있다.

Identification of circulating transcripts associated with adenoma-carcinoma sequence in the colorectal cancer

김리아 Graduate School, Yonsei University 2024 국내박사

ABSTRACT Identification of Circulating Transcripts Associated with Adenoma-Carcinoma Sequence in the Colorectal Cancer Li Ah KIM Department of Biomedical Laboratory Science The Graduate School Yonsei University Colorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer-related mortality. The conventional model for CRC progression traces its origins from normal epithelial cells through non-advanced and advanced adenomas, ultimately culminating in cancer. While this model has been instrumental in colorectal cancer surveillance, its reliance on tissue sampling presents challenges in accurately determining cancer stages. This study addresses these limitations by identifying circulating transcriptomic biomarkers associated with the adenoma-carcinoma sequence in CRC, utilizing readily available blood samples for increased screening efficacy. Blood samples were obtained from 160 individuals scheduled for colonoscopy who were systematically divided into five groups based on both colonoscopy and histology results: CRC group, advanced adenoma (AA) group, non-advanced adenoma (NA) group, symptomatic non-disease control (NDC) group, and healthy control (HC) group. RNA sequencing was performed on 20 samples from each group, and the subsequent results were subjected to three gene ontology analysis methods. Further analysis included network examination using heat map, protein-protein interaction, and KEGG pathway analyses. The study yielded remarkable results, identifying IFI27 in the NDC group, DEFA4 in the NA group, MPO in the AA group and CD177 in the CRC group as significant circulating transcriptional biomarkers. These biomarkers, which are closely linked to immune responses, suggest the involvement of immune cells, particularly neutrophils, in CRC progression. Of particular importance is the marked upregulation of myeloperoxidase (MPO) in the NA and AA groups compared to the HC group, a validation confirmed by RT-qPCR testing on a subset of 60 blood samples. The implications of this research underscore the potential utility of specific circulating transcriptomic biomarkers associated with the adenoma-carcinoma sequence for the early detection and management of CRC. This innovative approach represents a non-invasive method to identify precancerous adenomas in the bloodstream, offering promising prospects for early intervention prior to tissue manifestation. Key words: colorectal cancer, adenoma-carcinoma sequence (ACS), RNA sequencing, circulating transcripts, biomarker, neutrophil, MPO, RT-qPCR 국문초록 대장암 선종-암화 기전과 관련된 순환 전사체의 식별 연세대학교 대학원 임상병리학과 김리아 대장암(CRC)은 전 세계적으로 세 번째로 흔한 암이며 암 관련 두 번째 주요 사망 원인이다. CRC 진행에 대한 선종-암화 기전은 정상 상피 세포에서 비진행성 선종과 진행성 선종을 거쳐 궁극적으로 암으로 발전하는 과정을 설명하고 있다. 이 기전은 대장암 감시에 중요한 역할을 했지만 조직 샘플링에 의존하기 때문에 실제 임상적으로 암 단계를 정확하게 결정하는데 어려움이 있다. 이 연구는 CRC의 선종-암화 기전과 관련된 순환 전사체 바이오마커를 식별하고, 암에 대한스크리닝 효능 증가를 위해 쉽게 이용 가능한 혈액 샘플을 활용함으로써 이러한 한계를 해결하고자 했다. 대장내시경 검사를 받기로 예정된 160명의 개인으로부터 혈액 샘플을 채취하여 대장내시경 검사와 조직학 결과를 바탕으로 총 5개의 그룹, CRC 그룹, 진행성 선종(AA) 그룹, 비진행성 선종(NA) 그룹, 증상이 있는 비질병 대조(NDC)그룹과 건강한 대조(HC)그룹 으로 체계적으로 분류하였다. 각 그룹당 20개 샘플, 총 100개의 샘플로 RNA sequencing을 수행하였고, 이후 결과를 3가지 유전자 온톨로지 분석 방법에 적용하였다. 추가 분석에는 히트 맵, 단백질-단백질 상호 작용 및 KEGG 경로 분석을 사용한 네트워크 연구를 시행하였다. 이 연구에서는 중요한 순환 전사체바이오마커로 NDC 그룹의 IFI27, NA 그룹의 DEFA4, AA 그룹의 MPO, CRC 그룹의 CD177을 선정할 수 있었다. 면역 반응과 밀접하게 연관되어 있는 이러한 바이오마커는 CRC 진행에 면역 세포, 특히 호중구가 관여함을 시사한다. 특히 중요한 것은 HC 그룹에 비해 NA 및 AA 그룹에서 골수과산화효소(MPO)의 현저한 상향 조절이 있는 것을 HC그룹과 NA, AA그룹에 해당하는 각 20개의 혈액 샘플, 총 60개의 혈액 샘플에 대하여 RT-qPCR 테스트를 통해 검증하였다. 혈액으로부터 순환 전사체 바이오마커를 이용하여 대장암의 진행과정을 식별하는 이러한 혁신적인 접근법은 전암성 선종을 진단하는 비침습적 방법을 제시하며, 조직에서의 암 발현 보다 앞서 혈액으로부터 순환 전사체 바이오마커를 검출함으로써 대장암에 대한 조기 진단과 예후 전략을 향상시키는데 유용함을 보여주고 있다. ________________________________________핵심되는말: 대장암, 선종-암종 서열, RNA 염기서열분석, 순환 전사체, 바이오마커, 호중구, MPO, RT-qPCR

우리나라 農産物 流通 構造의 調整에 관한 硏究

김용국 明知大學校 國際通商大學院 1996 국내석사

As the reason for need to improve the circulating structure for the agricultural products of Korea is to enumerate the elements such as 1) The growth of circulating margins by the complication of the circulating processes, 2) The insufficiency of the authority's systematically rearing policy on the marketplace, 3) The inefficient roles of the agricultural cooperative-union in the place of production market, 4) The lack of marketplace information for the producer's price, 5) The price information that can not be reflected in the disharmony of the demand and supply. The purpose of researching this thesis is to present the improving proposals for the circulating structure in connection with the agricultural products. The method of this study based on the analysis for the circulating structure of agricultural products keeping pace with the kind of reports, monthly investigating magazines and statistical materials. The extend of this study is as following 1. The theoretical survey on the circulating structures and processes of the agricultural products market. 2. The analysis on the circulating margins of agricultural products. 3. The analysis on the conditions of the demand and supply for agricultural products. And so, the conclusion of this study is as following : 1. It is needed to be strengthen the circulating function in the place of products market for the agricultural products. That is to say, it is needed to strengthen the function such as a joint collections through the joint forwards, a assortment packing. It must be establish, activate the agriculture's spontaneous cooperate-system and the huge mechanism-collections specially the place of products, the products, and so it is needed to be spread the direct transactions with the central wholesale placemarket, the large retail placemarket. 2. The Authority, the agricultural cooperate-unions, the agriculture's organization is needed to be role faithfully in the circulating systems. Therefore, the authority should try to endeavour the price-control through the public price-collusion by the market-auction system, all kinds of the information's and the propaganda. The agricultural cooperate-unions should strengthen the functions such as the merchant functions, the collection functions and dissolution functions. The unit agricultural's organization should organically relate with the unit agriculture cooperate-unions. On the basis of this conclusions, for the purpose of improving the circulating structures of agricultural products, the authority must establish the department taking sole charge of circulating prices, the circulating placemarketsand the statistics, should try to active the establishment for market-auction system. Through the this system, the consumers should settle the organic relation with the authority, the agricultural cooperat-unions, the agriculture through the using fur's agricultural products. Afterwards, when the authority establish the agricultural policy i.e.., the improvement of the circulating structures of agricultural products, if it refer to this theory, it is thought to be profit. Our cemetery system is based on traditional Korean system. The Korean people are deeply attached to the worship of ancestor spirits. This worship is derived from the tenets of confucius and grounded within the regional peculiarities of our country. The elaborate funerals process and grove selection of our elders stem from the sense of filial duty and respect held. Because of this, the choice of propitious grove site for family members is of paramount importance, although we hold our customs dears, we also now consider the needs of our large population and the needs of effective land resource use. The dignified burial of our decal versus the land needs of the living causes concern. Currently, 921.8 square kms of land is held as cemetery (18,220,000 grove sites). This equates to approximately 0.9% of the total land moss. 10.5 square kms of land (210,000 grove sites) ore added each sear, on the average. The increase con be described as adding the land moss of skidoo in Seoul 1.3 times per sear. This has grate impact on the korean environment and land use policies for the growing population. Additional statistics ore equally sobering. The rote of increase for cemetery land in relation to the total land area will be 1.1 % in another 2000yrs. Government figures on the rate of increase for crematoriums are : 1984(16.6%), 1986(16.4%), 1987(17.8%). Korean population density is 12 times the world average. This density is the 3rd highest in the world. More seriously, the amount of cemetery land compared with total land is the highest in the world. Thus one may say the land is virtually covert with cemeteries. Because of this situation, our cemetery system should be improved. A program with which to alleviate this problem is presented 1. There have been no problems with current desire program to limit growth of the amounts of land in cemeteries from 2. squire meters to 10 square meters per grave sites. This may be gradually enforced as time goes on. 2. A system of imposing a time limit for burial may be introduced gradually. The limit would permit exhumation squire reburial in another cemetery location as land is needed. This is tone in France, where cremation follows a 3-30 year burial. This would permit a customary a burial permit, exhumation e cremation then interment for paying respects in a charnel house. 3. Legislation may be introduced which permits cremation, supports it administratively e prohibit burial of the dead. 4. Cemetery taxes can be newly established, with exception of public national cemeteries. This tax would be imposed to provide for the use of the nations land as well as maintenance a public crematorium facility a limited public cemetery. Illegal burials should be examined e discouraged by imposing a system of heavy tastes which may be levied. Also cemetery "Collectivization" e unauthorized occupation of the land holdings for cemeteries should be prohibited by tat lazy revisions. 5. The above suggestions will not be effective unless the public is educated on cemetery use, e cemetery legislation needs The previous cemetery improvement measures pan only be effective if there is consensus on retiring those pustoms which are archaic e will not serve future generation. Changing our beliefs, howervez, should be ,done gradually e with great foresight, if the new program are to be acceptable. It is possible that the dead refuge pan become a park enjoyed by living for refreshment e contemplation These should be are goals in order to pass both respect to our ancestors e a better environment to our dependants.

Identification of circulating miRNAs as diagnostic biomarkers of hair cell damage in sensorineural hearing loss

하선목 Graduate School, Yonsei University 2024 국내박사

Sensorineural hearing loss (SNHL) is caused by damage to the inner ear cells, specifically, the hair cells, which convert sound signals into bioelectrical signals. Early diagnosis is crucial to prevent damage to the inner ear cells, which can result in permanent hearing loss. Circulating microRNAs (miRNAs) are small RNAs that are released from cells and circulate in the blood. The quantity and variety of circulating miRNAs depend on the cell's state. Therefore, this study aimed to identify circulating miRNAs that can aid in diagnosing hair cell damage in SNHL. For this study, first, cochlear were isolated from 3-day-old mice to identify circulating miRNAs associated with hair cell damage. The hair cells were then exposed to culture media directly through ex vivo culture. To induce hair cell damage, the cochlear explant was treated with kanamycin, an ototoxic reagent. Subsequently, miRNAs released from the cells into the culture media were investigated. For this study, 8 miRNAs (miR-183, miR-205, miR-15a, miR-18b, miR-23a, miR-34a, miR-143, and miR-210), previously identified as having an association with SNHL, were selected and tested as hair cell damage biomarkers. In summary, the study found that kanamycin treatment of cochlear explants resulted in a statistically significant increase in miR-183, miR-18b, and miR-210, and a decrease in miR-23a expression levels. Additionally, the study compared miRNA levels in the plasma of patients with idiopathic sudden sensorineural hearing loss (ISSHNL) without other underlying diseases with those of healthy subjects. In conclusion, the study revealed a statistically significant increase in miR-183 and miR-210 levels and a decrease in miR-18b, miR-23a and miR-143 levels. The study suggests that circulating miRNA biomarkers, specifically miR-183, miR-18b, miR-23a, and miR-210, have a diagnostic value in identifying hair cell damage in SNHL. The sensitivity and specificity of these biomarkers were found to be 81.0% and 87.5%, respectively. This indicates the potential for developing a diagnostic method for hair cell damage in SNHL using these circulating miRNA biomarkers. 감각신경성 난청은 소리 신호를 생체 전기 신호로 바꾸는 내이 세포, 특히 유모세포의 손상 때문에 발생하는 질환이다. 내이 세포의 손상은 영구적인 청력 손실을 초래할 수 있어, 이를 조기에 진단하는 것이 중요하다. 본 연구는 감각신경성 난청에서 유모세포 손상을 진단하는 데 유용한 마이크로 RNA 를 규명하는 것을 목표로 하였다. 우선, 유모세포 손상과 관련된 순환 마이크로 RNA 를 확인하기 위해 생후 3 일 된 마우스의 달팽이관을 분리하여 생체 외에서 배양하고, 유모세포를 배양액에 직접 노출시켰다. 이후, 달팽이관 외이식편에 이 독소인 카나마이신을 처리하여 유모세포 손상을 유도한 후, 세포에서 배양액으로 방출되는 마이크로 RNA 를 조사하였다. 본 연구에서는 이전 연구에서 감각신경성 난청과 연관성이 확인된 8 개의 마이크로 RNA(miR-183, miR-205, miR15a, miR-18b, miR-23a, miR-34a, miR-143, 그리고 miR-210)를 유모세포 손상의 바이오 마커로 선별하여 테스트하였다. 연구 결과, 달팽이관 외이식편에 카나마이신 처리를 통하여 통계적으로 유의미한 수준에서 방출된 miR-183, miR-18b, miR-210 의 증가와 miR-23a 의 감소를 확인하였다. 이 상관관계를 더 강화하기 위해, 감각신경성 난청 중 기저 질환이 없는 돌발성 난청이 있는 환자의 혈장 마이크로 RNA 발현 수준을 건강한 사람과 비교하였다. 결과적으로, miR-183 와 miR-210 수치는 유의미하게 증가하고, miR-23a, miR-18b, miR-143 수치는 감소하였다. 또한, miR-183, miR-18b, miR-23a, miR-210 의 순환 마이크로 RNA 바이오마커를 이용한 진단은 각각 81.0%와 87.5%의 민감도와 특이도를 확인하였다. 이러한 결과는 순환 마이크로 RNA 바이오마커를 활용한 감각신경성 난청의 유모세포 손상의 진단법 개발의 잠재성을 규명하였다.