• 대한수의학회지
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• 제36권2호
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• pp.277-281
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• 1996

A technique to improve the analysis of micronuclei(MN) in lymphocytes as a cytogenetic indicator is reported. For the purpose of diminishing the variation of the result from individual reader and making it easier to distinguish accurately a cytokinesis blicked(CB) lymphocyte and micronuclei, we tried a modified one-step silver staining technique as a method for detection of the argyrophilic nucleolar organizer region(AgNOR) with or without conventional Giemsa stain in the slide from CB method. Compared with the conventional Giemsa stain, the preparation processed with this method are especially useful for the accurate analysis of MN of cultured lymphocyte with cytochalasin B. This method will be a useful technique for automated calculation of MN.

• Tuberculosis and Respiratory Diseases
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• 제42권4호
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• pp.513-521
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• 1995

연구배경: 세포의 증식능력을 반영한다고 알려진 NORs를 간편한 은염색으로 발현시켜서 AgNORs 수가 정상조직과 종양조직의 비교감별과 종양의 증식능력판단에 유용한지를 알기위하여 파라핀 포매된 편평세포폐암 조직을 이용하여 본 연구를 시행하였다. 방법: 수술로써 절제한 파라핀 포매된 편평세포폐암 조직 36예를 Mourad 등의 방법으로 은염색하였다. 1,000 배 배율하에서 종양조직과 정상조직에서 임의로 100 개씩 세포를 선정해서 핵당 평균 AgNORs수를 구하였다. 결과: 종양조직에서 TNM병기에 따른 핵당 평균 AgNORs수는 유의한 차이가 없었으나 정상조직과 종양조직을 비교허여 정상조직에서는 $1.74{\pm}0.25$, 종양 조직에서는 $4.05{\pm}0.80$으로 종양조직의 핵당 평균 AgNORs수가 정상조직에 비해서 유의하게 높았다(p<0.001). 결론: TNM분류에 따른 각 병기별 종양조직과 인접 정상조직의 핵당 평균 AgNORs수를 비교하여도 각 병기에서 종양조직의 평균 AgNORs수가 정상조직에 비하여 유의하게 높았다(p<0.005).

• Toxicological Research
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• 제14권4호
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• pp.489-494
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• 1998

In the present study, we have evaluated cytotoxic effects of Taraxaci Herba extract on human skin melanoma cells. The light microscopic study showed morphological changes AG-NOR (argyrophylic nucleolar organizer region) by silver chloride stain, and glycoprotein by PAS reaction of the treated cells. Disruptions in cell organelles were determined by SRB assay.

• 대한의생명과학회지
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• 제10권4호
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• pp.361-366
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• 2004

The purpose of this study was to determine the effect of high voltage pulsed current (HVPC) stimulation on proliferative activities of basal keratinocytes by measured nucleolar organizer region (NOR) expression and thickness of spinous layer in aged rat skin. Fifty-one weeks old twelve male Sprague-Dawley rats (300∼350g) were divided into control and HVPC stimulation groups. Each animal's hair on the back were removed. The HVPC stimulation group received an negative monophasic twin peak pulsed current stimulation with 50 V, while the control group was given the same treatment without electricity. The rats were sacrificed after 3 weeks. The biopsy specimens were fixed in formalin, embedded in paraffin and stained with hematoxyline-eosin and silver nitrate. The thickness of basal to granular layer of the epidennis were measured using a light microscope and computerized image analysis system. The number of argyrophilic nucleolar organizer region (AgNOR) were counted using a light microscope and computerized image analysis system and calculated as the mean number of AgNOR per nucleus in the basal keratinocyte. By using a Student's t-test, an increase in the thickness of basal-spinous layer (P<0.001) of epidermis can be observed in HVPC stimulation rats as compared with the control rats, whereas the thickness of the granular layer is not affected. A Student's t-test showed a significantly higher mean NOR number per nucleus of the basal keratinocyte in the HVPC stimulation rats than control rats (P<0.001). There was significantly positive correlation between the NOR number and the thickness of basal-spinous layer (r=0.80, P<0.05). These results suggest that the HVPC stimulation may increase the thickness of spinous layer in the epidennis due to increased proliferative activities of basal keratinocytes in epidennis in aged rat skin.

• The Journal of Korean Physical Therapy
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• 제12권1호
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• pp.1-7
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• 2000

전기자극이 흰쥐 창상 표피세포의 핵소체형성부위에 미치는 영향을 규명할 목적으로 Sprague-Dawley계 수컷 흰쥐 24마리의 등애 15mm의 절개창을 만들고 봉합한 후 각각 4일 7일 대조군 및 전기자극에 각각 6마리씩 무작위로 배치하고 전기자극군 흰쥐의 에 전극을 부착한 후 맥동빈도 0.5pps, 전류량 $500{\mu}A$로 60분간 전기자극하였으며 대조군은 전극만 부착하고 전류를 통전시키지 않았다. 전자자극 후 등의 피부를 적출하여 포르말린에 고정한 후 파라핀 포매 절편을 제작하여 silver colliod염색을 하였다. 이 표본을 400배율 현미경하에서 관찰하여 핵당 AgNOR으 수를 계측한 후 핵당 AgNOR의 분포와 핵당 평균 AgNOR수를 산출하여 통계분석한 결과 창상유발 4일과 7일째의 전기자극군의 평균 AgNOR수가 대조군보다 통계학적으로 유의하게 높게 나타났다. 이러한 결과는 전기자극이 흰쥐 피부 창상연 표피세포에서 AgNOR 발현을 증가시켰음을 확인할 수 있었으며 이는 미세맥동전류자극이 창상연의 표피세포를 증식시키고 있음을 시사하고 있다.

• 한국환경성돌연변이발암원학회지
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• 제18권2호
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• pp.104-108
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• 1998

In the present study, we have evaluated cytotoxic effects of Taraxaci Herba extract in human oral epitheloid carcinoma cells. An antitumor activity was measured by colorimetric assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulforhodamine protein B (SRB). The light microscopic study showed morphological changes, Ag-NOR (argyrophylic nucleolar organizer region) number and PAS positive reaction or the treated cells. These results obtained are as follows : MTT and SRB quantities were significantly decreased in cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations. The number of Ag-NORs were significantly decreased in cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations and the rate of Ag-NORs was shifted to left side (one Ag-Nounucleus was increased and five Ag-NORs/nucleus were decreased) by the high concentration. PAS reaction of cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations was negative. These results suggest that Taraxaci Herba retains a potential antitumor activity.

• 약학회지
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• 제43권2호
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• pp.173-179
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• 1999

In the present study, we have evaluated cytotoxic effects, antitumor activities and metastasis inhibitory effects of Palsun brewing water in NIH 3T3 cells, human epitheloid carcinoma cells, and human skin melanoma cells. The light microscopic study showed morphological changes, AG-NOR (argyrophylic nucleolar organizer region) by silver chloride stain, and glycoprotein by PAS (periodic acid stain) reaction of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide) and SRB (sulforhodamine B protein) assays. These results suggest that Palsun Brewing Water retains no cytotoxic effects in NIH 3T3 cells and a growth-inhibitory activity in cancer cell lines.

• 대한수의학회지
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• 제30권4호
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• pp.459-464
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• 1990

Inhibitory effect of caffeine on NIH3T3 cell proliferation was studied by using [$^3H$]thymidine incorporation and a modified one-step silver staining technique. The latter technique shows argyrophilic nucleolar organizer region-associated proteins (Ag-NORs), which are seen in nuclei as black dots. The result was compared with the counts of [$^3H$] thymidine incorporation. The results were summarized as follows; 1. The Ag-NORs numbers of NIH3T3 cells were $6.81{\pm}1.38$ at 24 hrs, $7.13{\pm}1.26$ at 48 hrs, $8.50{\pm}2.04$ at 72 hrs after incubation. 2. The numbers of Ag-NORs were significantly decrease in caffeine treated groups (p<0.01). 3. The counts of [$^3H$] thymidine incorporation were similar to the result of using Ag-NORs staining technique. It is concluded that Ag-NOR is a rapid, simple and compatible method to quantitate cell proliferation as compared with [$^3H$]thymidine incorporation.