• Molecular & Cellular Toxicology
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• 제1권4호
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• pp.256-261
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• 2005

The recent DNA microarray technology enables us to understand gene expression profiling in cell line and animal models. The technology has potential possibility to comprehend mechanism of multiple genes were related to compounds which have toxicity in biological system. So, microarray system has been used for the prediction of toxicity through gene expression induced by toxicants. It has been shown that compounds with similar toxic mechanisms produce similar changes in gene expression in vivo system. Here we focus on the use of toxicogenomics for the determination of gene expression analysis associated with hepatotoxicity in rat liver and cell line (WB-F344). Methotrexate (MTX) is a chemotherapy agent that has been used for many years in the treatment of cancer because it affects cells that are rapidly dividing. Also it has been known the toxicity of MTX, in a MTX abortion, it stops embryonic cells from dividing and multiplying and is a non-surgical method of ending pregnancy in its early stages. We have shown DNA microarray analyses to assess MTX-specific expression profiles in vivo and in vitro. Male Sprague-Dawely VAF+ albino rats of 5-6 weeks old and WB-F344 cell line have been treated with MTX. Total RNA was isolated from Rat liver and cell line that has treated with MTX. 4.8 K cDNA microarray in house has been used for gene expression profiling of MTX treatment. We have found quite distinct gene expression patterns induced by MTX in a cell line and in vivo system.

• Journal of Ginseng Research
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• 제27권2호
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• pp.47-51
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• 2003

Sarcoma 180 암세포를 이식한 마우스에서 면역조절 작용을 갖는 홍삼산성다당체 (RGAP)를 항암제 cyclophosphamide (CY)와 병용투여한 결과, RGAP (100 mg/kg)와 CY (3 mg/kg)의 병용투여는 고용량의 CY (10 mg/kg) 단독투여보다 강한 항암작용을 나타내었다. 또한 RGAP (100 mg/kg)과 CY (10 또는 20 mg/kg)과의 병용투여는 CY의 단독투여보다 LL/2 폐종양을 보다 강하게 억제하였다. RGAP (100 mg/kg) 와 5-fluorouracil (5-FU) (2.5 mg/kg)을 병용투여하였을 때도 sarcoma 180 암세포를 이식한 마우스에서 5-FU 단독투여군에 비해 현저한 항암시너지 효과를 나타내었다. LL2 폐 종양에 미치는 결과에서도 RGAP (100 mg/kg)와 5-FU (5 또는 10 mg/kg) 병용투여군은 항암제 단독투여군보다 강한 항종양 효과를 나타내었다. 이상의 결과로부터 RGAP는 항암제 CY 또는 5-FU와 병용투여시 sarcoma 180 및 LL/2 폐종양에 대해서 저용량 항암제의 사용으로 고용량의 항암제 사용에서와 유사한 항암효과를 나타낼 수 있어서 항암효과를 극대화 시킬뿐만아니라 항암제 사용에 의한 면역독성과 같은 부작용을 경감시킬 수 있는 항암치료 보조제로서의 사용가능성을 제시하였다.

• Korean Journal of Radiology
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• 제22권3호
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• pp.476-488
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• 2021

Objective: We aimed to develop a deep neural network for segmenting lung parenchyma with extensive pathological conditions on non-contrast chest computed tomography (CT) images. Materials and Methods: Thin-section non-contrast chest CT images from 203 patients (115 males, 88 females; age range, 31-89 years) between January 2017 and May 2017 were included in the study, of which 150 cases had extensive lung parenchymal disease involving more than 40% of the parenchymal area. Parenchymal diseases included interstitial lung disease (ILD), emphysema, nontuberculous mycobacterial lung disease, tuberculous destroyed lung, pneumonia, lung cancer, and other diseases. Five experienced radiologists manually drew the margin of the lungs, slice by slice, on CT images. The dataset used to develop the network consisted of 157 cases for training, 20 cases for development, and 26 cases for internal validation. Two-dimensional (2D) U-Net and three-dimensional (3D) U-Net models were used for the task. The network was trained to segment the lung parenchyma as a whole and segment the right and left lung separately. The University Hospitals of Geneva ILD dataset, which contained high-resolution CT images of ILD, was used for external validation. Results: The Dice similarity coefficients for internal validation were 99.6 ± 0.3% (2D U-Net whole lung model), 99.5 ± 0.3% (2D U-Net separate lung model), 99.4 ± 0.5% (3D U-Net whole lung model), and 99.4 ± 0.5% (3D U-Net separate lung model). The Dice similarity coefficients for the external validation dataset were 98.4 ± 1.0% (2D U-Net whole lung model) and 98.4 ± 1.0% (2D U-Net separate lung model). In 31 cases, where the extent of ILD was larger than 75% of the lung parenchymal area, the Dice similarity coefficients were 97.9 ± 1.3% (2D U-Net whole lung model) and 98.0 ± 1.2% (2D U-Net separate lung model). Conclusion: The deep neural network achieved excellent performance in automatically delineating the boundaries of lung parenchyma with extensive pathological conditions on non-contrast chest CT images.

• IMMUNE NETWORK
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• 제22권3호
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• pp.22.1-22.25
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• 2022

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndromecoronavirus-2 (SARS-CoV-2), has spread over the world causing a pandemic which is still ongoing since its emergence in late 2019. A great amount of effort has been devoted to understanding the pathogenesis of COVID-19 with the hope of developing better therapeutic strategies. Transcriptome analysis using technologies such as RNA sequencing became a commonly used approach in study of host immune responses to SARS-CoV-2. Although substantial amount of information can be gathered from transcriptome analysis, different analysis tools used in these studies may lead to conclusions that differ dramatically from each other. Here, we re-analyzed four RNA-sequencing datasets of COVID-19 samples including human bronchoalveolar lavage fluid, nasopharyngeal swabs, lung biopsy and hACE2 transgenic mice using the same standardized method. The results showed that common features of COVID-19 include upregulation of chemokines including CCL2, CXCL1, and CXCL10, inflammatory cytokine IL-1β and alarmin S100A8/S100A9, which are associated with dysregulated innate immunity marked by abundant neutrophil and mast cell accumulation. Downregulation of chemokine receptor genes that are associated with impaired adaptive immunity such as lymphopenia is another common feather of COVID-19 observed. In addition, a few interferon-stimulated genes but no type I IFN genes were identified to be enriched in COVID-19 samples compared to their respective control in these datasets. These features are in line with results from single-cell RNA sequencing studies in the field. Therefore, our re-analysis of the RNA-seq datasets revealed common features of dysregulated immune responses to SARS-CoV-2 and shed light to the pathogenesis of COVID-19.

• Biomolecules & Therapeutics
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• 제32권4호
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• pp.481-491
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• 2024

Paxlovid is the first approved oral treatment for coronavirus disease 2019 and includes nirmatrelvir, a protease inhibitor targeting the main protease (M^pro) of SARS-CoV-2, as one of the key components. While some specific mutations emerged in M^pro were revealed to significantly reduce viral susceptibility to nirmatrelvir in vitro, there is no report regarding resistance to nirmatrelvir in patients and animal models for SARS-CoV-2 infection yet. We recently developed xenograft tumors derived from Calu-3 cells in immunodeficient mice and demonstrated extended replication of SARS-CoV-2 in the tumors. In this study, we investigated the effect of nirmatrelvir administration on SARS-CoV-2 replication. Treatment with nirmatrelvir after virus infection significantly reduced the replication of the parental SARS-CoV-2 and SARS-CoV-2 Omicron at 5 days post-infection (dpi). However, the virus titers were completely recovered at the time points of 15 and 30 dpi. The virus genomes in the tumors at 30 dpi were analyzed to investigate whether nirmatrelvir-resistant mutant viruses had emerged during the extended replication of SARS-CoV-2. Various mutations in several genes including ORF1ab, ORF3a, ORF7a, ORF7b, ORF8, and N occurred in the SARS-CoV-2 genome; however, no mutations were induced in the M^pro sequence by a single round of nirmatrelvir treatment, and none were observed even after two rounds of treatment. The parental SARS-CoV-2 and its sublineage isolates showed similar IC₅₀ values of nirmatrelvir in Vero E6 cells. Therefore, it is probable that inducing viral resistance to nirmatrelvir in vivo is challenging differently from in vitro passage.

• Toxicological Research
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• 제8권2호
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

• 대한골관절종양학회지
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• 제4권1호
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• pp.13-21
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• 1998

Taxol이 악성 골종양 세포에 어느 정도의 세포독성이 있는지를 평가하기 위해 한국세포주 은행에서 분양 받은 G-292, SaOS-2 및 HT-1080의 3가지 악성 골종양 세포들을 대상으로 기존의 항암제인 methotrexate, adriamycin, ifosfamide, cisplatinum과 함께 각각 투여하여 MTT분석법으로 정량 및 비교 분석하였으며, adriamycin과 taxol을 병용 투여하여 항암제의 상호작용을 isobologram 분석법으로 조사하여 다음과 같은 결과를 얻었다. 1. Taxol의 악성 골종양 세포 주들에 대한 $IC_{50}$는 G-292에서는 $2.7{\times}10^{-2}{\mu}g/ml$, $SaOS^{-2}$에서는 $1.0{\times}10^{-2}{\mu}g/ml$, $HT{\times}1080$에서는 $1.1{\times}10^{-3}{\mu}g/ml$이었다. 2. Taxol은 악성 골종양 세포주들에 대해 기존의 항암제들 보다 강한 세포독성을 보였으며, 기존 항암제의 세포 독성의 강도는 adriamycin이 제일 높은 역가를 보였으며 그 외 methotrexate, cisplatinum, ifosfamide순이었다. 3. Taxol과 adriamycin을 병용 투여하여 상호작용을 관찰한 결과 G-292와 SaOS-2 세포 주에서 상승효과가 관찰되었으며, HT-1080에서는 상승효과가 관찰되지 않았다.

• Journal of Hospice and Palliative Care
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• 제22권3호
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• pp.105-116
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• 2019

목적: 소아청소년 호스피스 완화의료(이하 소아완화의료)는 생명을 위협하는 질환을 가진 소아청소년의 삶의 질을 향상시기 위한 총체적 돌봄 철학이자 실무의 표준이다. 국내에서는 2018년 7월 국가 지원의 소아완화의료 시범사업을 도입하였는데, 소아완화의료의 발전 방향을 제시하기 위해 국외 선진국의 소아완화의료 제공체계를 고찰하였다. 방법: 소아완화의료 제공 수준을 검토하여 영국, 미국, 일본, 싱가포르를 대상으로 선정하였다. 소아완화의료 제공체계를 다룬 국내외 학술지 등 문헌을 통합적으로 고찰하고 현지 전문가의 자문과 현지 방문조사를 수행하였다. 1990년 이후 영어, 일본어로 발간된 문헌을 중심으로 PubMed, Google, Google Scholar에서 검색하여 학술지, 정책보고서 등을 참고하였다. 각국의 소아완화의료 발전과정, 정책, 재정 모델, 대상 기준, 전달 체계, 질 관리 체계에 대해 분석하였다. 결과: 영국은 지역사회의 독립형 소아전문 완화의료기관이 일차 의료체계와 협력하며 어린이병원의 전문 소아완화의료 자문팀과 의뢰와 자문을 주고받는다. 미국은 병원기반의 전문 소아완화의료 자문팀을 중심으로 지역사회의 호스피스기관, 가정의료기관이 네트워크를 구축하고 돌봄 코디네이터를 지정하여 소아완화의료를 제공한다. 일본은 완화의료, 재택의료, 장애아동 및 만성질환아동 지원체계에서 완화의료 성격의 서비스를 제공한다. 싱가포르는 소아전문 가정 완화의료 단체가 어린이병원의 전문 자문팀과 협력하여 높은 지리적 접근성을 토대로 중추적으로 완화의료를 제공한다. 결론: 국외의 제공체계를 참고하고 국내의 현장의 요구를 반영하여 소아완화의료의 제공체계를 정비하여 미충족 요구가 발생하지 않도록 한국 소아청소년 호스피스완화의료를 최적화해 나가야 한다.

• 한국의학물리학회지:의학물리
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• 제25권3호
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• pp.139-142
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• 2014

본 연구의 목적은 흡수선량 재구성, 방사선 치료간의 재구성된 선량의 등록, 선량-체적 히스토그램의 생산등을 수행하는 선량 재구성의 임상적 응용시스템을 만들고 그것을 변형된 전립선 팬텀에 적용하는 것이다. 이를 위해 변형가능한 전립선 팬텀을 20 cm 깊이와 40 cm너비의 물팬텀에 집에 넣었다. 이것의 영상을 얻고, 전립선, 정낭 및 항문의 윤곽을 그렸다. 동일 평면에서 네개의 조사문을 이용하여 세기 변조계획을 세웠다.항문에 20 ml의 물풍선을 삽입하여 장기를 변형시켰다. 영상을 다시 획득하여 위 장기의 윤곽을 그렸다. XVMC몬테칼로 코드를 사용하여 두 팬텀및 EPID내에서 선량반응 인자를 계산하였다. 세기변조계획에서 얻어진 방사선을 두팬텀에 조사하여 EPID에서 적분형 영상을 얻었다. Demons 방법을 사용하여 변형된 팬텀을 변형전 팬텀에 등록시켰다. 이를 통해 단위체적별 위치변이 정보를 얻었고 이를 이용해 두 팬텀의 재 구성된 선량을 합하여 변형전 팬텀에 생산해 냈다. 순방향으로 계산된 치료계획 선량을 합산된 재구성된 선량과 비교하였다. 200 cGy에서 전립선과 정낭이 받든 체적은 차이를 거의 보이지 않았으나, 210 cGy 이상에서는 3%가량 차이를 보였다. 항문에서는 150-200 cGy영역에서 재구성된 선량에 의하여 받은 체적은 치료 계획과 비교하여 3% 이상 적었다. 본 연구를 통하여 선량 재구성의 임상적 응용시스템이 성공적으로 만들어 졌다. 변형된 전립선 팬텀에 적용되어 작지 않은 선량의 차이를 목표장기와 보호 장기에 보였다.

• 생명과학회지
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• 제28권8호
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• pp.945-954
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• 2018

오메가-3 지방산(오메가-3)은 수종의 암에 대해 종양형성 억제 및 침윤이 억제됨이 알려져 있다. 그러나 혀의 편평세포암 세포에서 내인성 오메가-3에 의한 침윤 및 종양형성 억제 대한 연구가 명확하게 보고된 바 없다. 이에 본 연구는 혀의 편평세포암 세포에서 ${\omega}3$-fatty acid desaturase의 유전자 발현이 침윤 및 종양형성에 미치는 영향을 규명하였다. 먼저 SCC-4 및 SCC-9세포의 침윤능은 오메가-3인 DHA 처리에 의해 억제 됨을 확인 하였다. DHA 처리 후 MMP-9 및 MMP-2 활성이 감소 되었을 뿐만 아니라 그 promoter의 reporter 활성도 억제하였다. 또한 COX-2 및 VEGF promoter 활성 뿐만 아니라 NF-kB 활성도 DHA에 의해 억제 되었다. SCC-9의 ${\omega}3$-desaturase 유전자 stable 세포(fSCC-9sc)의 세포증식 및 colony formation이 억제 되었으며, in vivo 동물실험에서 fSCC-9sc 세포의 종양형성능은 현저히 억제 되었고, 면역형광염색법을 이용한 fSCC-9sc 세포의 종양 조직에서의 TUNEL 양성세포는 대조군인 fSCC-9cc 세포에 비해 현저히 증가하였다. 이상의 결과로 오메가-3는 인체 혀의 편평세포암 세포의 침윤 뿐만 아니라 종양형성을 억제하여 항암작용을 나타낼 수 있으며 따라서 오메가-3는 인체 혀의 편평암의 예방 및 치료에 유용하게 사용될 수 있으리라 생각된다.