• 생명과학회지
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• 제26권12호
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• pp.1458-1465
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• 2016

본 연구는 oxidative stress에 의한 세포죽음 분석의 이상적인 모델로 사용되는 HT22세포를 이용하여 천문동 에탄올추출물의 glutamate에 의한 oxidative toxicity에 대한 신경보호 효과를 살펴보았다. 이를 위해 cell viability, lactate dehydrogenase (LDH), 그리고 세포죽음형태, reactive oxygen species (ROS), mitochondria membrane potential (MMP) 등에 대한 flow cytometry 및 Western blot분석을 이용하였다. 천문동 추출물의 처리는 cell viability 및 LDH분석에서 glutamate에 의한 cell toxicity를 저하시키며, 특히 apoptotic cell death를 현저히 감소시켰다. ROS 및 MMP분석 결과, 천문동 추출물은 ROS의 형성을 저하시키며 glutamate에 의해 저하된 MMP를 현저히 회복시켜 주었다. 이와 관련된 단백질 발현을 보면, 천문동 추출물은 PARP 및 HO-1의 발현을 억제하였다. 이상의 결과는 천문동 추출물이 HT22해마세포에서 ROS형성저해 및 MMP회복에 의해 세포죽음을 완화시켜 보호작용을 하는 것으로 사료되며 oxidative toxicity관련 질환에 적용 가능할 것으로 보여 진다.

• 한국수산과학회지
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• 제34권1호
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• pp.17-20
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• 2001

한국산 꺽지과 어류 3종에 대한 세포유전학적 기초 자료를 얻기 위하여 염색체 핵형, 적혈구 세포 및 핵의 크기와 DNA 함량을 조사하였다 꺽지 (Coreoperca herzi), 꺽저기 (C. kawamebari) 및 쏘가리 (Siniperca schezeri)의 핵형은 각각 2n=48 (4SM+44A, T), 2n=48 (6SM+42A, T) 2n=48 (4SM+44A, T)이었으며, 암수간 heteromorphic한 성염색체는 발견할 수 없었다. 쏘가리의 적혈구 세포 및 핵의 표면적과 체적은 다른 2종보다 작았다. DNA함량은 적혈구 세포 크기에서 같은 경향을 보여, 쏘가리가 1.47pg/cell로 꺽지 (1.83pg/cell) 및 꺽저기 (1.85pg/cell)보다 유의하게 작았다.

• 한국ITS학회 논문지
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• 제10권1호
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• pp.16-26
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• 2011

가변속도제어(Variable Speed Limit ; VSL)는 교통상황에 따라 속도를 차등 제한하여 혼잡 또는 사고지점에 도달하기전 차량간, 차로간 속도편차를 감소시킴으로써 사고를 예방하고 총 지체를 최소화하기 위한 연속류 ITS기법 중 하나이다. 본 연구에서는 가변속도 제어시 연속류 기본구간의 동적교통류에 대한 수학적 분석결과를 기초로 가변속도제어 수행모형을 제시하고 효과분석을 수행하였다. 가변속도제어모형은 병목현상 발생시 상류부에 전파되는 충격파의 속도를 최소화 함으로써 지체를 감소시킬 수 있도록 graphical solution을 적용하여 제시한다. 또한 가변속도 제어시 해당 지점으로부터 하류부로 전파되는 충격파의 생성 및 움직임을 모형화 하여 재현하며 그 방법론으로는 Cell-Transmission Model 및 Supply-Demand Method를 적용하였다. 가변속도제어 적용에 따른 효과분석으로는 제어를 통한 지체감소의 효용성 및 안전도 측면에서 각 Cell별 속도 및 밀도의 표준편차의 개선정도를 살펴보았다. 향후 본 연구의 이론적 검증 및 분석 결과를 토대로 다양한 가변속도제어모형 및 운영방안을 개발할 수 있을 것으로 예상한다.

• 한국약용작물학회지
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• 제17권5호
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• pp.341-345
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• 2009

Cerebral ischemia results from a transient or permanent reduction in cerebral blood flow that decreases oxygen and glucose supply. When the cellular oxygen supply is reduced to critical level, damage to cells and induction of cell death are occurred by excitotoxicity, oxidative stress and inflammation. Ischemia remains one of the leading causes of death, but there is no effective treatment that might protect neurons gainst ischemia by interrupting the cascade of cell death. In this study, human neuroblastoma SH-SY5Y cells are exposed to oxygen and glucose deprivation (OGD) followed by reoxgenation. OGD can mimic the acute restriction of metabolite and oxygen supply caused by ischemia and is widely used as a model of ischemic conditions. SH-SY5Y cells are treated samples at the commencement of OGD to achieve different final concentrations, and cell viabilities were quantified using the measurement of flow cytometry analysis. Of those tested, the extracts of Polygala tenuifolia (roots), Dictamnus dasycarpus (barks), Polygala tenuifolia (roots), Eucommia ulmoides (branches), Eucommia ulmoides (barks), Poria cocos (whole), Sophora flavescens (roots) showed neuroprotective effects, with $EC_{50}$ values of $4.5{\pm}0.6$, $7.9{\pm}1.5$, $10.5{\pm}0.7$, $18.4{\pm}1.9$, $19.6{\pm}0.3$, $21.6{\pm}1.9$, and $30.7{\pm}3.9{\mu}g/m{\ell}$, respectively.

• 대한한방내과학회지
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• 제35권3호
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• pp.317-331
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• 2014

Objectives: The root of Platycodon grandiflorum (PG) has been known to possess a range of pharmacological activities including anti-cancer, anti-inflammatory, and anti-oxidant effects. The present study was designed to investigate whether or not PG-induced cell death was connected with autophagy and apoptosis in NCI-H460 human lung cancer cells. Methods: Effects on the cell viability and apoptotic activity were quantified using MTT assays and flow cytometry analysis, respectively. Protein activation was measured by immunoblotting. Autophagy was measured by LC3 immunofluorescence and immunoblotting. ROS production and loss of mitochondria membrane potential (MMP) were checked with flow cytometry analysis. Results: Following exposure to PG, NCI-H460 cell proliferation decreased simultaneously inducing autophagic vacuoles and up-regulation of microtubule-associated protein 1 light chain 3 and beclin-1 protein expressions. Interestingly, pre-treated with autophagy inhibitors, 3-methyladenin or bafilomycin A1 further triggered reduction of cell viability. PG treatment also induced apoptosis that was related modulation of Bcl-2 family proteins, death receptors and activation of caspases. In addition, PG stimulation clearly enhanced loss of MMP and reactive oxygen species (ROS) generation. Conclusions: Our results suggest that PG elicited both autophagy and apoptosis by increasing loss of MMP and ROS production. PG induced-autophagy may play a cell protective role.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9945-9948
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• 2014

Background: The purpose of this study was to investigate Tim-3 expression on peripheral CD3-CD56+ natural killer (NK) cells and CD3+CD56+ natural killer T (NKT) cells in lung cancer patients. Materials and Methods: We analyzed Tim-3+CD3-CD56+ cells, Tim-3+CD3-$CD56^{dim}$ cells, Tim-3+CD3-$CD56^{bright}$ cells, and Tim-3+CD3+CD56+ cells in fresh peripheral blood from 79 lung cancer cases preoperatively and 53 healthy controls by flow cytometry. Postoperative blood samples were also analyzed from 21 members of the lung cancer patient cohort. Results: It was showed that expression of Tim-3 was significantly increased on CD3-CD56+ cells, CD3-$CD56^{dim}$ cells and CD3+CD56+ cells in lung cancer patients as compared to healthy controls (p=0.03, p=0.03 and p=0.04, respectively). When analyzing Tim-3 expression with cancer progression, results revealed more elevated Tim-3 expression in CD3-CD56+ cells, CD3-$CD56^{dim}$ cells and CD3+CD56+ cells in cases with advanced stages (III/IV) than those with stage I and II (p=0.02, p=0.04 and p=0.01, respectively). In addition, Tim-3 expression was significantly reduced on after surgical resection of the primary tumor (p<0.01). Conclusions: Tim-3 expression in natural killer cells from fresh peripheral blood may provide a useful indicator of disease progression of lung cancer. Furthermore, it was indicated that Tim-3 might be as a therapeutic target.

• Asian Pacific Journal of Cancer Prevention
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• 제16권6호
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• pp.2555-2559
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• 2015

Purpose: To investigate effects of the TESTIN (TES) gene on proliferation and migration of highly metastatic nasopharyngeal carcinoma cell line 5-8F and the related mechanisms. Materials and Methods: The target gene of human nasopharyngeal carcinoma cell line 5-8F was amplified by PCR and cloned into the empty plasmid pEGFP-N1 to construct a eukaryotic expression vector pEGFP-N1-TES. This was then transfected into 5-8F cells. MTT assays, flow cytometry and scratch wound tests were used to detect the proliferation and migration of transfected 5-8F cells. Results: A cell model with stable and high expression of TES gene was successfully established. MTT assays showed that the OD value of 5-8F/TES cells was markedly lower than that of 5-8F/GFP cells and 5-8F cells (p<0.05). Flow cytometry showed that the apoptosis rate of 5-8F/TES cells was prominently increased compared with 5-8F/GFP cells and 5-8F cells (p<0.05). In vitro scratch wound assays showed that, the width of the wound area of 5-8F/TES cells narrowed slightly, while the width of the wound area of 5-8F/ GFP cells and 5-8F cells narrowed sharply, suggesting that the TES overexpression could inhibit the migration ability. Conclusions: TES gene expression remarkably inhibits the proliferation of human nasopharyngeal carcinoma cell line 5-8F and reduces its migration in vitro. Thus, it may be a potential tumor suppressor gene for nasopharyngeal carcinoma.

• 대한한방부인과학회지
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• 제21권2호
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• pp.27-37
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• 2008

Purpose: This study was conducted to investigate the effects of Ginseng Radix Alba extract solution on the cell cycle regulation, apoptosis of human uterine myoma cells. Methods: Primary cultured human uterine myoma cells were treated with extract solution of Ginseng Radix Alba concentration of 1, 10 and $100mg/m{\ell}$ for 48 hours. We underwent flow cytometry and western blotting for cell cycle and apoptosis related factors. Results: In flow cytometry, a slight promotion of G1 phase in $1mg/m{\ell}$ group had been observed but, simultaneously a delay of G1 phase in 10 and $100mg/m{\ell}$ groups were also observed. The manifestation of cyclin D1 in Ginseng Radix Alba extract solution medicated group increased compared to the control group. The manifestation of Bax controlling apoptosis increased in terms of concentration but Bcl-2 showed no change. Also VEGF increased in terms of concentration. Conclusion: The present study suggests that Ginseng Radix Alba extract solution treatment in human uterine myoma cells induce the delay of cell cycle and apoptosis. These results suggest that Ginseng Radix Alba will be a promising agent for use in therapeutics agents against human uterine myoma.

• 대한수의학회지
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• 제50권3호
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• pp.179-185
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• 2010

The present study was undertaken to establish reference values for the composition blood lymphocyte populations and compare forty three Hanwoo neonatal calves (KC) with twenty one Holstein calves (HC) by blood cell count and immunophynotying. The percentages of CD2+, CD4+, CD8+, CD26+, ACT2+, MHC class, MHC class II and WC1+ T cells, B cells were determined by flow cytometry. The number of lymphocyte and monocyte in HC were higher than those of KC. However, the number of neutrophils was higher in HC than KC. The proportions of CD2+, CD4+, CD8+, MHC class, and WC1+ lymphocytes remained relatively stable during the study period, while there was a moderate increase in the relative percentage of CD26+, ACT2+, MHC class II and B cell from birth to approximately 3 weeks of age. Marked differences in the relative proportions of the lymphocyte subpopulations were noted between the individual calves. The present study shows that the T-cell subpopulations are present in peripheral blood of KC at levels comparable with HC, while the MHC class II and B cell population of KC increases significantly with age. The absolute number of WBC in KC was due to the decrease of absolute number of neutrophil rather than the increase of lymphocyte. The results indicated that KC have significantly higher number of neutrophils, and proportion of MHC class II and B cell than HC.

• IMMUNE NETWORK
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• 제6권3호
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• pp.138-144
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• 2006

Background: There are at least two different subsets of B cells, B-1 and B-2. The characteristic features and function of B-2 cells in addition to the effect of steroids on B-2 cells are well-known. Although B-1 cells have different features and functions from B-2 cells, the effect of steroids on B-1 cells is not completely understood. Therefore, this study examined the effects of dexamethasone on peritoneal (or B-1 cells) and splenic B cells (or B-2 cells). Methods: Purified B cells were obtained from the peritoneal fluid and the spleens of mice. The isolated B cells were cultured in a medium and after adding different concentrations of dexamaethasone. The cell survival rate was measured by flow cytometry using propidium iodide. The expression level of the B cell surface marker was analyzed by flow cytometry. During the culture of these cells, immunoglobulin secreted into the culture supernatants was evaluated by an enzyme-linked immunosorbent assay. Results: The survival rate of peritoneal and splenic B cells decreased with increasing dexamethasone concentration. However, the rate of peritofieal B cell apoptosis was lower than that of splenic B cells. CDS and B7.1 expression in peritoneal B cells and CD23 and sIgM expression in splenic B cells after the dexamethasone treatment were reduced. When B cells were treated with dexamethasone, the spontaneous IgM secretion decreased with increasing dexamethasone concentration. Conclusion: Dexamethasone induces apoptosis in peritoneal and splenic B cells. However, peritoneal B cells are less sensitive to dexamethasone. The dexamethasone suppressed expression of the surface markers in peritoneal B cells is different from those in splenic B cells.