실험 방광암 모델에 있어서 Proliferating Cell Nuclear Antigen이 발현되는 세포들의 색상-화상 분석

이현무,이태수,김원재,차은종,송형근 충북대학교 의과대학 충북대학교 의학연구소 1993 忠北醫大學術誌 Vol.3 No.1

Proliferating cell nuclear antigen(PCNA)에 대한 단클론성 항체에 의해 면역조직화학적으로 염색된 세포들의 평가에 있어서 색상-화상 분석의 역할을 살펴보고자 하였다. BBN 투여 기간에 따라 Fisher 344 백서 방광에 정상 상피로부터 단순 과증식, 결절성 혹은 유두상 과증식 및 표재성 방광암에 이르는 순차적인 병리세포학적 변화가 발생함을 관찰한 후, 이를 색상-화상 분석과 육안적 분석을 통해 PCNA에 대한 평균 labeling index를 구한 결과 두가지 방법에서 모두 각 병변 사이에 통계적으로 유의한 차이를 보였으나, 색상-화상 분석을 이용하여 구한 평균 labeling index의 변이계수가 육안적 분석을 이용하여 구한 평균 labeling index의 변이계수에 비해 전 병변에 대하여 낮게 나타나, 종양화과정이 진행함에 따라 PCNA에 대한 평균 labeling index가 증가하고 평균 labeling index가 높을 수록 증식도가 더 큼을 나타낸다는 결과를 얻었으며, 색상-화상 분석을 이용하여 얻은 결과가 육안적 분석을 이용하여 얻은 결과보다 신뢰도가 더 높은 것으로 나타났다. 이와같은 면역조직화학염색에 대한 색상-화상 분석을 통한 세포역동학적 연구는 정량화 및 재현성의 향상을 통해 면역조직화학염색의 평가에 있어 보다 신뢰도가 높은 정보를 제공하여줄 것이며, 이를 통해 방광종양의 증식도 및 악성도를 예측하는데 중요한 역할을 할 것으로 생각된다. A study was carried out to define the role of color-image analysis in evaluation of immunohistochemically labeled cells using anti-PCNA/cyclin monoclonal antibody. Sequential cellular changes(normal, simple hyperplasia, nodular or papillary hyperplasia, and transitional cell carcinoma-Ta, T1) were observed in Fisher 344 rat urinary bladder depending on the duration of BBN administration. The mean PCNA labeling index obtained by color-image analysis and visual analysis in normal, simple hyperplasia, nodular or papillary hyperplasia and bladder tumor showed statistically significant difference in each histological group. But coefficients of variation of mean PCNA labeling index obtained by color-image analysis were lower than those using visual analysis in all lesions. Higher PCNA labeling indices indicated greater biological malignant potential, and mean labeling index in each histological group obtained by both color-image and visual analysis progressively increased with tumorigenesis, but the coefficient of variation with color-image analysis was lower than that with visual analysis in general, therefore, the evaluation of labeling indices obtained using color-image analysis was more reliable than that obtained using visual analysis. The data suggest that cell kinetic evaluation using color-image analysis may provide more reliable information to evaluate immunohistochemical staining in terms of quantitation and reproducibility and play an important role in predicting proliferative activity and malignant potential for bladder tumor.

Lifespan Extending and Stress Resistant Properties of Vitexin from Vigna angularis in Caenorhabditis elegans

( Eun Byeol Lee ),( Jun Hyeong Kim ),( Youn-soo Cha ),( Mina Kim ),( Seuk Bo Song ),( Dong Seok Cha ),( Hoon Jeon ),( Jae Soon Eun ),( Sooncheon Han ),( Dae Keun Kim ) 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.6

Several theories emphasize that aging is closely related to oxidative stress and disease. The formation of excess ROS can lead to DNA damage and the acceleration of aging. Vigna angularis is one of the important medicinal plants in Korea. We isolated vitexin from V. angularis and elucidated the lifespan-extending effect of vitexin using the Caenorhabditis elegans model system. Vitexin showed potent lifespan extensive activity and it elevated the survival rates of nematodes against the stressful environments including heat and oxidative conditions. In addition, our results showed that vitexin was able to elevate antioxidant enzyme activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner. These studies demonstrated that the increased stress tolerance of vitexin-mediated nematode could be attributed to increased expressions of stress resistance proteins such as superoxide dismutase (SOD-3) and heat shock protein (HSP-16.2). In this work, we also studied whether vitexin-mediated longevity activity was associated with aging-related factors such as progeny, food intake, growth and movement. The data revealed that these factors were not affected by vitexin treatment except movement. Vitexin treatment improved the body movement of aged nematode, suggesting vitexin affects healthspan as well as lifespan of nematode. These results suggest that vitexin might be a probable candidate which could extend the human lifespan.

Soy Sauce Lowers Body Weight and Fat Mass in High-Fat Diet-Induced Obese Rats

Cha Youn-Soo,Jung Eun Park,Eun-Ji Lee,Jeongwoo Song,Chan-Ho Park,Eun-Gyung Mun,Jinxi Wang,한안나 한국식품영양과학회 2023 Journal of medicinal food Vol.26 No.11

Soy sauce (SS) is a traditional fermented seasoning. Although fermented foods have diverse health beneficialeffects, SS intake has been discouraged because of its high salt level. This study was designed to evaluate the antiobesityoutcomes of SS and the potential involvement of salt content in SS by adding a high-salt group. Sprague-Dawley rats wererandomly assigned into four groups: normal diet (ND, 10% fat of total kcal), high-fat diet (HD, 60% fat of total kcal), HD withsalt water (HDSW, NaCl = 8%), and HD with SS (HDSS, NaCl = 8%). SS significantly decreased HD-induced body weightgain and lipogenic gene expression without affecting food consumption. Moreover, SS also reduced hepatic injury and lipidaccumulation, and also improved hyperlipidemia. Furthermore, SS decreased the mRNA levels related to obesity-derivedinflammatory responses, while HDSW did not change the levels of those markers. These observations indicate that SSameliorates obesity in HD-fed obese rats by attenuating dyslipidemia. Moreover, SS might also have an anti-inflammatoryeffect in HD-induced obesity, which requires further investigation. Most importantly, SS offers these beneficial effectsregardless of its high salt content, implying that different dietary salt sources lead to the distinct health outcomes. Inconclusion, the findings of this study improve the understanding of the functional effect of SS.

Lifespan Extending and Stress Resistant Properties of Vitexin from Vigna angularis in Caenorhabditis elegans

Lee, Eun Byeol,Kim, Jun Hyeong,Cha, Youn-Soo,Kim, Mina,Song, Seuk Bo,Cha, Dong Seok,Jeon, Hoon,Eun, Jae Soon,Han, Sooncheon,Kim, Dae Keun The Korean Society of Applied Pharmacology 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.6

Several theories emphasize that aging is closely related to oxidative stress and disease. The formation of excess ROS can lead to DNA damage and the acceleration of aging. Vigna angularis is one of the important medicinal plants in Korea. We isolated vitexin from V. angularis and elucidated the lifespan-extending effect of vitexin using the Caenorhabditis elegans model system. Vitexin showed potent lifespan extensive activity and it elevated the survival rates of nematodes against the stressful environments including heat and oxidative conditions. In addition, our results showed that vitexin was able to elevate antioxidant enzyme activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner. These studies demonstrated that the increased stress tolerance of vitexin-mediated nematode could be attributed to increased expressions of stress resistance proteins such as superoxide dismutase (SOD-3) and heat shock protein (HSP-16.2). In this work, we also studied whether vitexin-mediated longevity activity was associated with aging-related factors such as progeny, food intake, growth and movement. The data revealed that these factors were not affected by vitexin treatment except movement. Vitexin treatment improved the body movement of aged nematode, suggesting vitexin affects healthspan as well as lifespan of nematode. These results suggest that vitexin might be a probable candidate which could extend the human lifespan.

전국 병원영양사의 직무만족도 현황 및 관련요인

장은희 ( Eun Hee Jang ),류은순 ( Eun Soon Lyu ),이송미 ( Song Mi Lee ),박유경 ( Yoo Kyung Park ),이은 ( Eun Lee ),차진아 ( Jin A Cha ),박미선 ( Mi Sun Park ),이호선 ( Ho Sun Lee ),라미용 ( Mi Young Rha ),이승민 ( Seung Min Lee ) 대한영양사협회 2014 대한영양사협회 학술지 Vol.20 No.4

The objective of this study was to examine job satisfaction status and its related factors among dietitians working in hospitals. A survey questionnaire was sent to dietitians working in tertiary, general, and convalescent hospitals in Korea. The study sample hospitals included all tertiary hospitals (n=43), and the sample of general hospitals (n=114) and convalescent hospitals (n=240) was randomly selected using a stratified sampling method. A total of 665 dietitians completed the survey questionnaire. The questionnaire included27 job satisfaction questions on task, stability·vision, working conditions, and relationship areas, and the questions on task improvement need. The job satisfaction scores in all four areas ranged from2.53/5.00 to 3.89/5.00, implying generally low job satisfaction level in hospital dietitians. The job satisfaction levels in the convalescent hospital group were significantly lower compared to those in the tertiary and general hospital groups. Whereas job satisfaction levels of tertiary and convalescent hospital groups did not differ by number of beds, those of the general hospital group increased significantly with the number of beds for all four areas. Multivariate regression analysis results showed that hospital type was a significant predictor of job satisfaction level for all four areas. Job satisfaction of relationship area was also significantly affected by the number of beds, whereas that of working conditions was influenced by annual income and working experience. The study findings provide useful information in planning clinical dietitians` optimal supply outlook for the improvement of clinical nutrition services.

Effects of Ranolazine on Norepinephrine-Induced Cell Death by Inhibition of the β-adrenoceptor Signal Pathway in Cardiomyocytes

( Kyung Eun Kim ),( Hee Sang Song ),( Hye Jung Kim ),( Min Ji Cha ),( Byeong Wook Song ),( Eun Ju Choi ),( On Ju Ham ),( Chang Yeon Lee ),( Seong Yong Choi ),( Se Yeon Lee ),( Yang Soo Jang ),( Tae Wo 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.12

Ranolazine blocks the intracellular sodium and calcium overload accompanying myocardial ischemia and is used in antianginal therapy. The effects of ranolazine on the β-adrenoceptor signal transduction system are poorly understood. The present study was designed to confirm whether the mechanism was associated with a β-adrenoceptor antagonist activity of ranolazine on norepinephrine (NE)-induced cardiomyocytes. After cardiomyocytes were preincubated with propranolol and ranolazine, cells were treated with NE for 24 hours. The phosphorylation of ERK was decreased by ranolazine treatment, in comparison with NE-only treated cells. Intracellular Ca2+and Na+ levels decreased by 40±3% and 17±0.5%, respectively, compared to control. Ranolazine decreased expression of Ca2+/ calmodulin-dependent protein kinase II (CaMKII) by 57±4%, the Na+-Ca2+exchanger (NCX) by 21±0.5%, and the ryanodine receptor 2 (RyR2) by 47±1.5%, compared with NE-only treated control cells. Ranolazine also increased expression of the L-type Ca2+channel (LTCC) by 48±3.5%, phospholamban (PLB) levels by 45±2%, and the sarcoplasmic reticulum Ca2+ATPase 2a (SERCA2a) by 36±0.5%, compared to levels in NE-only stimulated cells. The number of annexin V/PI-positive cells fell by 39±1.5% after ranolazine treatment, compared with levels in NE-only treated control cells. Ranolazine also inhibited apoptosis by regulating the levels of the pro-apoptotic factor Bax, the anti-apoptotic factor Bcl-2, and cytochrome C release. These results demonstrate that ranolazine had an effect on NE-induced cell death through inhibition of the β-adrenoceptor signal pathway in cardiomyocytes.

CT-Derived Deep Learning- Based Quantification of Body Composition Associated with Disease Severity in Chronic Obstructive Pulmonary Disease

Jae Eun Song,So Hyeon Bak,Myoung-Nam Lim,Eun Ju Lee,Yoon Ki Cha,Hyun Jung Yoon,Woo Jin Kim 대한영상의학회 2023 대한영상의학회지 Vol.84 No.5

Purpose Our study aimed to evaluate the association between automated quantified body composition on CT and pulmonary function or quantitative lung features in patients with chronic obstructive pulmonary disease (COPD). Materials and Methods A total of 290 patients with COPD were enrolled in this study. The volume of muscle and subcutaneous fat, area of muscle and subcutaneous fat at T12, and bone attenuation at T12 were obtained from chest CT using a deep learning-based body segmentation algorithm. Parametric response mapping-derived emphysema (PRMemph), PRM-derived functional small airway disease (PRMfSAD), and airway wall thickness (AWT)-Pi10 were quantitatively assessed. The association between body composition and outcomes was evaluated using Pearson’s correlation analysis. Results The volume and area of muscle and subcutaneous fat were negatively associated with PRMemph and PRMfSAD (p < 0.05). Bone density at T12 was negatively associated with PRMemph (r = -0.1828, p = 0.002). The volume and area of subcutaneous fat and bone density at T12 were positively correlated with AWT-Pi10 (r = 0.1287, p = 0.030; r = 0.1668, p = 0.005; r = 0.1279, p = 0.031). However, muscle volume was negatively correlated with the AWT-Pi10 (r = -0.1966, p = 0.001). Muscle volume was significantly associated with pulmonary function (p < 0.001). Conclusion Body composition, automatically assessed using chest CT, is associated with the phenotype and severity of COPD.

Significance of microvascular evaluation of ductal lesions on breast ultrasonography: Influence on diagnostic performance

Kim, Eun Sil,Seo, Bo Kyoung,Park, Eun Kyung,Woo, Ok Hee,Jung, Kyoonsoon,Cho, Kyu Ran,Song, Sung Eun,Cha, Jaehyung Elsevier 2018 Clinical imaging Vol.51 No.-

<P><B>Abstract</B></P> <P>We aim to investigate the significance of microvascular ultrasonography in breast ductal lesions for distinguishing benign from malignant lesions. Sixty-two ductal lesions were evaluated with gray-scale and three Doppler techniques before biopsy, Superb Microvascular Imaging (SMI), color Doppler (CDI), and power Doppler imaging (PDI). We evaluated number, distribution, and morphology of vessels. The area under the ROC curve of combined use of Doppler imaging was greater than that of gray-scale only and SMI had higher value than CDI and PDI (<I>p</I> < 0.05). Microvascular ultrasonography improves diagnostic performance that distinguishes benign from malignant breast ductal lesions, and SMI is superior to CDI and PDI.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Breast ductal lesions are caused by various benign and malignant diseases. </LI> <LI> Gray-scale ultrasonogrpahy is limited in assessing ductal lesions because secretion can be confused with intraluminal tumor. </LI> <LI> We report the usefulness of microvascular ultrasonography to differentiate benign from malignant ductal lesions. </LI> <LI> Combined use of Doppler ultrasonography significantly improves diagnostic performance compared to gray-scale only. </LI> <LI> Superb Microvascular Imaging is superior to color or power Doppler technique in microvascular evaluation of ductal lesions. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

신생 쥐의 저산소성 허혈성 뇌손상에서 Nitric oxide synthase 조절을 통한 taurine의 신경보호 효과

송은정 ( Eun Jeong Song ),장윤영 ( Yoon Yung Jang ),김태열 ( Tae Yeol Kim ),차선화 ( Sun Hwa Cha ),박혜진 ( Hye Jin Park ),이계향 ( Kye Hyang Lee ),이경훈 ( Kyung Hoon Lee ),최은진 ( Eun Jin Choi ),김진경 ( Jin Kyung Kim ),정혜리 대한주산의학회 2009 Perinatology Vol.20 No.3

목적: 타우린은 술폰 기를 산기로 하는 황 아미노산의 일종으로 뇌, 망막, 심장, 근육에 풍부하여 담즙산의 포합 기능 뿐아니라 두뇌 발달, 망막의 광수용체 활성, 심장 근육의 수축, 삼투압 조절, 생식기능, 성장발달, 면역 체계의 유지 및 황산화 활성 등 다양한 생물학적 기능이 보고되면서 그 중요성이 재조명되고 있다. 최근 이러한 여러 가지 작용 중에서 국소적 뇌허혈에 대한 타우린의 신경보호효과에 관한 연구들이 발표되었다. 그러나 이러한 연구가 성인의 뇌졸중의 치료에 대한 연구에서 진행된 것이 대부분이며 아직까지 신생아시기의 저산소성 손상에 대한 효과를 구체적으로 연구한 바가 없어, 본 연구에서는 타우린이 발달하고 있는 뇌에서 NOS 조절을 통해 저산소성 허혈성 뇌손상을 줄여줄 수 있는지 알아보고자 하였다. 방법: 재태기간 18일된 태아 백서의 대뇌피질 세포를 배양하여 1% O2 배양기에서 저산소 상태로 뇌세포 손상을 유도하여 저산소군, 손상 전후 타우린 투여군(30 ug/mL)으로 나누어 정상산소군과 비교하였다. 또한, 생후 7일된 백서의 좌측 총 경동맥을 결찰한 후 저산소(8% O2) 상태로 2시간 노출시켜서, 저산소성 허혈성 뇌 손상을 유발하였고, 뇌손상 전후 30분에 타우린을 체중 kg당 30 mg을 투여하였다. 저산소성 허혈성 뇌손상 후 1일, 3일, 7일, 14일, 28일 째 뇌를 적출하여 질소산화물과 관련된 inducible NOS (iNOS), endothelial NOS (eNOS) 및 neuronal NOS (nNOS)을 이용하여 실시간 중합효소연쇄반응과 western blotting을 하였다. 결과: 저산소로 유발된 뇌세포 배양에서 정상군에 비해 저산소군에서 뇌세포 손상이 많았고 저산소 손상 전 타우린 투여군에서 뇌세포 손상이 회복되었으며 저산소 손상후 타우린 투여군에서는 저산소 손상 전 투여군보다 회복력이 떨어졌다. 실시간 중합효소연쇄반응과 western blotting을 이용한 저산소 상태의 백서 뇌세포 배양 실험뿐만 아니라 저산소성 허혈성 뇌손상 동물 모델에서도 타우린을 투여한 경우 저산소군에서는 손상 후 7일과 14일 째 iNOS는 감소하였고 eNOS와 nNOS는 증가하였으나 뇌 손상 전 타우린 투여군에서는 반대로 iNOS는 증가하였고 eNOS와 nNOS는 감소하였다. 결론: 본 연구에서 저산소 상태에서 발현된 iNOS 감소, eNOS와 nNOS 증가가 타우린 투여 후에는 반대로 정상 상태로 유지되는 것을 보아 질소 산화물 조절 기전을 통한 뇌보호 작용도 있을 것이라 생각된다. 그리고 이것은 대체적으로 저산소 손상 후 7일과 14일 째 가장 효과가 있는 것으로 밝혀졌다. Purpose: Taurine is a simple sulfur-containing amino acid and enriched in brain, retina, heart and skeletal muscles. In the central nervous system, taurine has been implicated in major phenomena. Current studies have demonstrated the neuroprotective effect of taurine in adult rat model, but limited data are available for those during the neonatal periods. The aim of this study was to determine whether taurine could reduce hypoxic-ischemic (HI) brain injury in the developing brain via modulation of nitric oxide synthase. Methods: In in vitro model, embryonic cortical neuronal cell culture procedure was done in Sprague-Dawley (SD) rats at 18 days of gestation. The cells were divided into the hypoxia group, taurine-treated group before and after a hypoxic insult. The each groups compared with normoxia group. In in vivo model, left carotid artery ligation was done in 7-day-old SD rat pups. the pups were exposed to hypoxia, received an injection of 30 mg/kg of taurine, and sacrificed at day 1, day 3, day 7, day 14 and day 28. We assayed the expression of iNOS, eNOS and nNOS mRNA using real-time PCR and western-blotting. Results: In in vitro model, brain cell damage of hypoxia group was more than in the normoxia group. Cell damage`s recovery was more in the taurine-treated group before a hypoxic insult than in the taurine-treated group after a hypoxic insult. The expression of iNOS mRNA was less in the hypoxia group than in the normoxia group both in vitro and in vivo models. The expression of eNOS and nNOS was more in the hypoxia group. Conclusion: Taurine has neuroprotective property over perinatal HI brain injury due to modulation of NOS, as evidenced by causing a decrease in eNOS and nNOS and increase in iNOS expression. The neuroprotective effect of taurine administration was maximal at day 7 and day 14 after a hypoxic injury.

Development of a Multiplex and Cost-Effective Genotype Test toward More Personalized Medicine for the Antiplatelet Drug Clopidogrel

Jeong, Hye-Eun,Lee, Su-Jun,Cha, Eun-Young,Kim, Eun-Young,Kim, Ho-Sook,Song, Young Hwan,Shin, Jae-Gook Molecular Diversity Preservation International (MD 2014 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.15 No.5

<P>There has been a wide range of inter-individual variations in platelet responses to clopidogrel. The variations in response to clopidogrel can be driven by genetic polymorphisms involved in the pathway of absorption, distribution, metabolism, excretion, and the target receptor P2Y12. A set of genetic variants known for causing variations in clopidogrel responses was selected, which included <I>CYP2C19*2</I>, <I>*3</I>, <I>*17</I>, <I>CYP2B6*4</I>, <I>*6</I>, <I>*9</I>, <I>CYP3A4*18</I>, <I>CYP3A5*3</I>, <I>MDR1 2677G</I> > <I>T</I>/<I>A</I>, <I>3435C</I> > <I>T</I>, and <I>P2Y12 H2</I> (<I>742T</I> > <I>C</I>). The simultaneous detection of these 10 variants was developed by using a multiplex PCR and single-base extension (MSSE) methodology. The newly developed genotyping test was confirmed by direct DNA sequencing in the representative positive control samples and validated in an extended set of 100 healthy Korean subjects. Genotyping results from the developed MSSE exhibited a perfect concordance with the direct DNA sequencing data and all of variants tested in 100 healthy Korean subjects were in agreement with Hardy-Weinberg equilibrium (<I>p</I> > 0.05). The present molecular diagnostic studies provide an accurate, convenient, and fast genotyping method for the detection of multiple variants. This would be helpful for researchers, as well as clinicians, to use genetic information toward more personalized medicine of clopidogrel and other antiplatelet drugs in the future.</P>