• Tuberculosis and Respiratory Diseases
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• 제42권3호
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• pp.314-321
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• 1995

연구배경: 종양세포는 세포의 비정상적인 분열성장이 증가되므로, 세포내의 DNA가 양적인 변화를 일으킨다. DNA의 양적변화인 DNA ploidy 여부는 종양의 생물학적 특성을 반영하므로, 소세포 폐임에서 DNA ploidy의 변화와 생존기간을 비교하였다. 방법: 1990년 1월부터 1991년 12월까지 원광의대 부속병원에서 원발성 소세포 폐암으로 조직병리학적 진단을 받고나서, 2회 이상의 화학요법을 실시받은 후, 최소 2년이상의 후향적 추적에 의해 사망이 확인된 42례를 대상으로 하였다. DNA ploidy 분석방법은 paraffin에 보관된 병리조직을 이용하여 유식세포 분석법에 의한 DNA histogram으로 분석하였다. DNA ploidy 여부와 평균 생존기간을 비교하였으며, 다시 TNM 병기, PS scale, 화학요법 실시 횟수 등으로 세분하여 DNA ploidy 여부와 생존기간과의 관계를 재비교하였다. 결과: 1) 전 군의 평균 생존기간은 190(${\pm}156$)일이었으며, TNM 병기, PS scale이 진행할 수록 생존기간은 단축되었다. 2) 전 군에서 DNA aneuploidy의 발현 비율은 60%(26/42)였으며, 암의 진행정도와는 관계없었다. 3) 전 군에서의 평균 생존기간은 diploidy군이 272(${\pm}197$)일로서 aneuploidy 군의 138(${\pm}90$)일보다 유의하게 연장되었다(p<0.001). 4) DNA ploidy 여부에 의한 생존기간의 차이에 대한 TNM 병기, PS scale의 영향은 없었다. 결론: 소세포 폐암 환자에서 DNA aneuploidy 군은 diploidy 군보다 유의하게 생존기간이 짧았으며, DNA ploidy 여부는 TNM 병기, PS scale과는 무관한 예후추정 인자로서 임상적 이용아 가능하다고 생각된다.

• Restorative Dentistry and Endodontics
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• 제22권1호
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• pp.374-387
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• 1997

The purpose of this study was to examine the distribution of lymphocyte populations in normal, reversibly inflamed and irreversibly inflamed human dental pulp tissues using flow cytometry. Flow cytometry, with specific antibody and fluorochrome reagent allows us to know cellular properties of hematolymphoid cells by measuring fluorescence of stained cells. Before extirpation of pulps in routine endodontic treatment, the clinical diagnosis were performed by symptom. The extirpated pulp tissues were divided into normal pulp group (N=5), reversible pulpit is group(N=10) and irreversible pulpitis group(N=7). The specimen was placed into RPMI 1640 medium, minced into small pieces, and then digested in medium with collagenase. The cell suspension was resuspended in PBS for monoclonal antibody staining of T lymhocytes(CD3+), B lymphocytes (CD19+), T helper cell (CD4+) and T supressor cell (CD8+). The percentages of cells were counted by FACStar(BD) flow cytometer. Following results were obtained; 1. In the most normal and inflamed pulps, the percentages of T lymphocyte, B lymphocytes, T helper cell and T suppressor/cytotoxic cell were less than 1 % in total counted pulpal cells. 2. The higher percentages of T, B, T helper and T suppressor cells were observed in irreversible pulpitis group as compared with the normal pulp and reversible pulpitis group but the differences between groups were not statistically significant (p>0.05). 3. The percentages of T helper cells (CD4 + cells) were greater than that of T suppressor/cytotoxic cells (CD8 + cells) in the inflamed pulps.

• Tuberculosis and Respiratory Diseases
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• 제56권5호
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• pp.514-522
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• 2004

연구 배경 : 비스테로이드성 항염증제는 대장암의 화학 예방에 이용되고 있다. 지속적으로 비스테로이드성 항염증제를 복용한 결과 대장암 발생의 위험이 40-50% 감소하였다. Sulindac은 비스테로이드성 항염증제의 일종으로 대장암의 예방 효과가 있으며 암세포의 성장 억제와 세포 고사를 유도시킨다. 이에 저자들은 3가지 비소세포 폐암 세포주에서 sulindac의 영향을 알아보고자 하였다. 방 법 : A549(선암), NCI-H157(편평상피암), NCI-H460(대세포암) 세포주에 sulindac을 농도별로 투여하여, MTT assay로서 암세포의 생존율을, 유식세포 분석법과 핵산 염색으로 세포 고사의 비율을, 유산탈수소효소유리로서 세포 사멸의 정도를 시간대별로 측정하였다. 결 과 : Sulindac에 의해 농도와 시간에 의존적으로 비소세포 폐암 세포주에서 암세포의 생존율이 감소하였고, 유산 탈수소 효소 유리는 증가하였으며, 세포 고사 역시 농도, 시간에 의존적으로 증가하였다. 결 론 : Sulindac은 비소세포 폐암 세포주에서 농도, 시간에 의존적으로 암세포의 생존율 감소와 세포 고사증가를 유도하였다.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1027-1033
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• 2007

Paclitaxel이 암세포에서 세포예정사를 유발할지라도, 아직 정확한 기전은 잘 알려져 있지 않다. 이에 본 연구에서는 HeLa $S_{3}$ 자궁암세포에서의 paclitaxel이 어떠한 영향을 미치는지 알아보고자 한다. 그리하여 방법으로는 세포독성검사, apoptotic cells의 형태학적 변화(DAPI 염색 ), western blot 분석법을 사용하여 수행하였다. 본 연구의 결과로 paclitaxel은 HeLa $S_{3}$ 세포에서 세포독성을 보이며 특히 paclitaxel의 $IC_{50}$ 값은 약 1 ${\mu}M$이며, paclitaxel 처리한 HeLa $S_{3}$ 세포에서 형태학적 변화(분절화)를 관찰하였고, flow cytometric 분석에서는 G2/M기가 차단되어 paclitaxel은 세포주기 특히 Sub-$G_{1}$기를 조절함을 알 수 있다. 그리고 Paclitaxel을 처리한 HeLa $S_{3}$ 세포에서는 PARP cleavage를 유발하였고 Bc1-2의 감소와도 관련되었다.

• 생명과학회지
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• 제24권1호
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• pp.92-97
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• 2014

Cordycepin은 Cordyceps militaris에서 처음 유래된 nucleoside adenosine 유도체의 일종으로 면역증강 및 항암활성을 포함한 다양한 약리 기능이 있는 것으로 알려져 있다. 본 연구에서는 LNCap 인체 전립선 암세포 모델을 이용하여 cordycepin에 의한 항암활성 기전을 연구하였다. Cordycepin 처리에 따라 LNCap 세포는 처리 농도 의존적으로 증식이 억제되었으며, 이는 apoptosis 유발과 연관성이 있음을 poly ADP-ribose polymerase의 단편화 현상과 Annexin V 염색에 의한 정량적 분석으로 확인하였다. Cordycepin 처리에 따른 flow cytometric analysis 결과로서 cordycepin이 세포주기 G2/M기 정체 현상을 유발하였음을 알 수 있었으며, 이는 cyclin B1 및 cyclin A의 발현 감소와 연관성이 있었다. 또한 cordycepin이 처리된 LNCap 세포에서 cyclin-dependent kinase (CDK) inhibitor p21Waf1/Cip1의 발현이 증가되었지만, CDK2, CDC2 및 Cdc25C의 발현에는 큰 영향을 미치지 않았으며, cordycepin에 의하여 증가된 p21 단백질은 CDK2 및 CDC2와의 복합체를 형성하고 있었다. 본 연구의 결과는 LNCap 전립선 암세포에서 cordycepin에 의한 G2/M 및 apoptosis 유발은 p53 비존적인 CDK inhibitor p21의 발현 증가가 중요한 역할을 하고 있음을 보여주는 것이다.

• 대한수의학회지
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• 제53권3호
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• pp.143-147
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• 2013

Ginsenoside Rp1 is one of ginseng saponins with chemotherapeutic activity. In this study, we investigated the effects of Rp1 on spleen cells. Spleen is a major immune organ consisted of crucial immune cells, such as T lymphocytes, B lymphocytes, natural killer cells, and some antigen-presenting cells. Although the anti-tumor potential of Rp1 was studied, the effects of Rp1 on immune cells have not investigated yet. A viability assay using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), flow cytometric analysis, Western blot analysis were used to detect cellular changes on Rp1-treated spleen cells. MTT assay showed that Rp1 decreased the viability of spleen cells. To further investigate the effects of Rp1 on activated spleen cells, we treated lipopolysaccharide (LPS) as a representative inflammatory agent and Rp1 on spleen cells in a combination. The surface expression levels of activation markers for lymphocytes, CD25 and CD69 were measured. Apoptotic analysis revealed the cytotoxic effects of Rp1 on both na$\ddot{i}$ve and activated cells, and the expression pattern of some apoptosis-related proteins was correlated to apoptotic events of cells. Taken together, ginsenoside Rp1 increases the cellular death of spleen cells and also inhibits the LPS-induced activation of spleen cells.

• Journal of Microbiology and Biotechnology
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• 제31권2호
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• pp.217-225
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• 2021

This study aimed to investigate the neuroprotective effects of 1-methoxylespeflorin G11 (MLG), a pterocarpan, against glutamate-induced neurotoxicity in neuronal HT22 hippocampal cells. The protective effects of MLG were evaluated using MTT assay and microscopic analysis. The extent of apoptosis was studied using flow cytometric analysis performed on the damaged cells probed with annexin V/propidium iodide. Moreover, mitochondrial reactive oxygen species (ROS) were assessed using flow cytometry through MitoSOXTM Red staining. To determine mitochondrial membrane potential, staining with tetramethylrhodamine and JC-1 was performed followed by flow cytometry. The results demonstrated that MLG attenuates glutamate-induced apoptosis in HT22 cells by inhibiting intracellular ROS generation and mitochondrial dysfunction. Additionally, MLG prevented glutamate-induced apoptotic pathway in HT22 cells through upregulation of Bcl-2 and downregulation of cleaved PARP-1, AIF, and phosphorylated MAPK cascades. In addition, MLG treatment induced HO-1 expression in HT22 cells. These results suggested that MLG exhibits neuroprotective effects against glutamate-induced neurotoxicity in neuronal HT22 cells by inhibiting oxidative stress and apoptosis.

• Journal of Microbiology and Biotechnology
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• 제33권1호
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• pp.106-113
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• 2023

The supply of microbiological risk-free water is essential to keep food safety and public hygiene. And removal, inactivation, and destruction of microorganisms in drinking water are key for ensuring safety in the food industry. Ultraviolet-C (UV-C) irradiation is an attractive method for efficient disinfection of water without generating toxicity and adversely affecting human health. In this study, the disinfection efficiencies of UV-C irradiation on Shigella flexneri (Gram negative) and Listeria monocytogenes (Gram positive) at various concentrations in drinking water were evaluated using a water purifier. Their morphological and physiological characteristics after UV-C irradiation were observed using fluorescence microscopy and flow cytometry combined with live/dead staining. UV-C irradiation (254 nm wavelength, irradiation dose: 40 mJ/cm²) at a water flow velocity of 3.4 L/min showed disinfection ability on both bacteria up to 10⁸ CFU/4 L. And flow cytometric analysis showed different physiological shift between S. flexneri and L. monocytogenes after UV-C irradiation, but no significant shift of morphology in both bacteria. In addition, each bacterium revealed different characteristics with time-course observation after UV-C irradiation: L. monocytogenes dramatically changed its physiological feature and seemed to reach maximum damage at 4 h and then recovered, whereas S. flexneri seemed to gradually die over time. This study revealed that UV-C irradiation of water purifiers is effective in disinfecting microbial contaminants in drinking water and provides basic information on bacterial features/responses after UV-C irradiation.

• Biomolecules & Therapeutics
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• 제18권4호
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• pp.420-425
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• 2010

The antifungal effects and action mechanisms of styraxjaponoside A and B were investigated. Devoid of hemolytic effect, the compounds had significant effect against several human pathogenic fungal strains, with energy-independent manners. To understand the action mechanisms of the compounds, the flow cytometric analysis plotting the forward scatter and the side scatter, $DiBAC_4$(3) staining and DPH fluorescence analysis were conducted. The results indicated that the actions of the compounds were dependent upon the membrane-active mechanisms. The present study suggests that styraxjaponoside A and B exert their antimicrobial effects via membrane-disruptive mechanisms.

• 대한한의학회지
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• 제26권4호
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• pp.12-21
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• 2005

Objectives: In the present study, we investigated whether an aqueous extract of Armeniacae semen induces apoptotic neuronal cell death upon mouse N2a neuroblastoma cells. Methods: 1. Cell viability was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTI) assay. 2. For in situ detection of apoptotic cells, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining. 3. The fraction of cells was revealed by flow cytometric analysis used that. 4. For detection of apoptotic DNA cleavage, DNA fragmentation assay was performed. 5. For detection of bax and bcl-2, Western blot analysis was performed. 6. Caspase enzyme activity was measured using caspase-3 assay. Results: From the present results, N2a neuroblastoma cells treated with Armeniacae semen extract exhibited several characteristics of apoptosis. A treatment of Armeniacae semen extract was shown to increase the expression of Bax, a proapoptotic protein, and the treatment decreased the expression of Blc2, an anti-apoptotic protein. In addition, Armeniacae semen extract increased the caspase-3 enzyme activity. Conclusions: The present results show that Armeniacae semen extract induces apoptotic cell death in mouse N2a neuroblastoma cells.