• ALGAE
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• 제25권4호
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• pp.197-204
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• 2010

Cytoskeletal changes were observed during cell division of the green alga Zygnema cruciatum using flourescein isothiocynate (FITC)-conjugated phallacidin for F-actin staining and FITC-anti-$\alpha$-tubulin for microtubule staining. Z. cruciatum was uninucleate with two star-shaped chloroplasts. Nuclear division and cell plate formation occurred prior to chloroplast division. Actin filaments appeared on the chromosome and nuclear surface during prophase, and the F-actin ring appeared as the cleavage furrow developed. FITC-phallacidin revealed that actin filaments were attached to the chromosomes during metaphase. The F-actin ring disappeared at late metaphase. At telophase, FITC-phallacidin staining of actin filaments disappeared. FITC-anti-$\alpha$-tubulin staining revealed that microtubules were arranged beneath the protoplasm during interphase and then localized on the nuclear region at prophase, and that the mitotic spindle was formed during metaphase. The microtubules appeared between dividing chloroplasts. The results indicate that a coordination of actin filaments and microtubules might be necessary for nuclear division and chromosome movement in Z. cruciatum.

• Clinical and Experimental Reproductive Medicine
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• 제46권2호
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• pp.60-66
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• 2019

Objective: To compare black rice (Oryza sativa L) extract with three different staining methods for human sperm head assessment. Methods: Semen samples were collected from 34 volunteers. Four smears of each ejaculate were prepared for staining using the rapid Papanicolaou (PAP) stain, SpermBlue, DipQuick, and black rice extract. The percentage of defective sperm heads (mean${\pm}$standard deviation) was compared. Results: Black glutinous rice extract, a natural dye, was used instead of hematoxylin to stain the nuclei of the sperm heads. The percentage of defective sperm heads showed a significant difference between black rice extract and DipQuick (p= 0.000). In contrast, black rice extract and rapid PAP showed no statistically significant difference (p= 0.974). A strong correlation (r = 0.761) was found between the findings obtained using rapid PAP and black rice extract. In contrast, a weak correlation (r = 0.248) was obtained between DipQuick and black rice extract for the percentage of defective sperm heads. Conclusion: The results showed good agreement and a strong correlation between the rapid PAP and black rice extract stains. The advantages of black rice extract as a novel substitute for hematoxylin for nuclear staining include ease of preparation, local availability, and favorable nuclear staining properties. Further studies could also focus on comparing staining techniques in clinical samples.

• 한국원자력학회:학술대회논문집
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• 한국원자력학회 2002년도 춘계공동학술발표회요약집
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• pp.388-388
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• 2002

• Journal of Radiation Protection and Research
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• 제29권1호
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• pp.9-15
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• 2004

방사선에 의해 유도되는 사람 말초혈액 림프구 미소핵 관찰빈도를 높이면서 생물학적 선량평가법으로서 활용 가능성을 확인하고자 본 연구를 수행하였다. 우선 5명의 건강한 사람으로부터 혈액을 제공 받아 선량영역을 0에서 800cGy로 하여 감마선$(^{137}Cs)$을 조사 한 후 김사와 아크리딘 오렌지 형광 염색하고 미소핵 출현빈도를 비교하였다. 아크리딘 오렌지 염색법을 이용하여 미소핵을 관찰하였을 때, 김사 염색법에 비교하여 적갈색의 비특이성 과립과 녹황색의 DNA가 붉은색의 세포질을 배경으로 명확히 구별되었을 뿐만 아니라 선량이 증가하면서 검출율도 높았다. 아울러, 말초혈액 T와 B-림프구에 대하여 선량영역을 0에서 50cGy로 하여 방사선을 조사한 후 미소핵 출현빈도를 아크리딘 오렌지 염색하여 김사염색과 비교한 바, B-림프구에서 선량이 증가하면서 적어도 2배 이상 높게 관찰되었다. 본 실험 결과, 사람 말초 혈액 B-림프구를 대상으로 한 아크리딘 오렌지 형광염색 미소핵 분석법은 저선량 방사선 인체영향 평가나 과피폭 선량추정시 활용이 가능 할 것으로 생각된다.

• 대한핵의학회지
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• 제38권6호
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• pp.511-515
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• 2004

목적: 갑상선세포에서 요오드의 섭취는 갑상선호르몬 합성의 첫 단계이며, sodium iodide symporter (NIS)라는 세포막 단백질에 의해 이루어지는 것으로 알려져 있고 NIS 유전자가 클로닝됨으로써 NIS 발현을 직접 관찰하는 것이 가능해졌다. 하지만, 갑상선암을 비롯한 갑상선 결절과 정상 조직에서의 NIS 발현은 검사방법과 대상에 따라 아주 다양한 결과를 보이고 있다. 따라서 이 연구에서는 갑상선암을 비롯한 여러 갑상선질환에서 NIS의 발현여부와 정도를 두 가지 서로 다른 RT-PCR방법과 면역조직화학염색법으로 조사하여 비교하였다. 대상 및 방법: 갑상선절제술을 받은 32명의 조직을 이용하였다. 술후 병리학적 진단은 유두암 19명, 여포암 1명, 수질암 1명, 선종 4명, 선종양 갑상선종 7명이었다. RT-PCR방법을 이용하여 갑상선글로불린, NIS, 갑상선과산화효소의 발현을 관찰하였고, 항NIS 항체를 이용하여 면역조직화학염색을 시행하여 NIS 발현 정도를 반정량적으로 평가하고, 그 결과를 각각 비교하였다. 결과: RT-PCR의 결과에서 유두암 19명중 10예에서 NIS의 발현이 있었다. 여포암 1예와 수질암 1예는 NIS 발현이 없었다. 선종 4명중 2예, 선종양 갑상선종 7명중 4예에서 NIS의 발현이 있었다. 면역조직화학염색법으로는 유두암 19명중 15예에서 발현이 있었고, 여포암 1예는 발현이 없었다. 선종 4예중 3예, 선종양 갑상선종 7예중 6예에서 발현이 있었다. RT-PCR방법에 의한 NIS의 발현 정도와 면역조직화학염색의 정도를 반정량적으로 평가하여 비교한 결과 각 검사의 결과 사이에 유의한 양의 상관관계가 있었다(p<0.001). 결론: RT-PCR방법과 면역조직화학염색법으로 조사한 NIS의 발현 정도는 양의 상관관계가 있었으나, RT-PCR방법에 비해 면역조직화학염색법으로 NIS발현을 더 많이 찾을 수 있었다.

• 대한물리치료과학회지
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• 제10권1호
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• pp.198-205
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• 2003

We evaluated the effect of low power GaAsAl laser on re-epithelization in full-thickness excisional wound of rat skin. Two full-thickness excisions were made on the back of the experimental animals. Low power laser applications with 10mW intensity were treated experimental animals twice a day for 7 days. On the seventh postoperative day the quantitative analysis of re-epithelization was performed using immunohistochemical staining for proliferating cell nuclear antigen (PCNA). The majority of PCNA immunoreactive cells was observed at epithelial cells in the margin of full thickness excisional wound. The low power laser treatments significantly increased the number of PCNA immunoreactive cell as compared to that of non treated animal group (p<0.01). The shape of PCNA immunoreactive cell appeared as small dark, round to ovoid structures. Most PCNA immunoreactive cells exhibited a high intensity of staining that contrasted sharply with the surrounding background. In conclusion, these findings suggest that GaAlAs laser treatments effectively enhance the epithelial wound healing by the stimulating cell proliferation. Furthermore, the majority of cell proliferation occurred in the margin of full thickness excisional wound.

• Asian Pacific Journal of Cancer Prevention
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• 제15권9호
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• pp.3959-3963
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• 2014

The tumour suppressor genes, p53 and pRb, are known to play important roles in neoplastic transformation. While molecular routes to the uncontrolled growth of hepatocytes, leading to primary liver cancer have generated considerable interest, the roles of p53 and pRb mutations in hepatocellular carcinoma (HCC) and hepatoblastoma (HB) remain to be clarified. We examined the immunohistochemical expression of p53 and pRb gene products in 26 HCC and 9 HB, sampled into tissue microarray blocks. 10 (38%) of 26 HCC showed > 10% tumour nuclear staining for p53 protein, 3 of these also being HbsAg positive. Conversely, none of 9 HB expressed nuclear p53 immunopositivity. Some 24 (92%) HCC and 8 (89%) HB showed loss of pRb nuclear expression. Two of the 26 HCC and one of the 9 HB showed >10% tumour nuclear staining for pRb protein. Our results suggest that p53 does not have an important role in the development of HB but may contribute in HCC. There is also loss of pRb expression in the majority of HCC and HB, supporting loss of pRb gene function in the hepatocarcinogenesis pathway. However, a comparison of the staining profiles of p53 and pRb proteins in HCC and HB did not reveal a consistent pattern to differentiate between the two types of tumours immunohistochemically. Hence the use of p53 and pRB protein expression has no contribution in the situation where there is a diagnostic difficulty in deciding between HCC and HB.

• Asian Pacific Journal of Cancer Prevention
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• 제13권8호
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• pp.3627-3630
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• 2012

The epithelial cell adhesion molecule (EpCAM) is a pan-epithelial differentiation antigen that is expressed on almost all carcinomas. However, a role in rat liver carcinogenesis has never been reported previously. Thus, its expression was investigated herein in rat liver tumors induced by diethylnitrosamine (DEN). Twenty male 5-week-old F344 rats were used in this experiment. Mini-osmotic pumps containing doses of 47.5 mg of DEN were inserted into the abdominal cavity of each animal to initiate liver carcinogenesis. All animals were sacrificed at 26 weeks after DEN treatment. At necropsy, hepatic masses were processed for histopathological examination, which revealed forty-four hepatocellular adenomas (HCAs) and twenty hepatocellular carcinomas (HCC). Tumors were immunohistochemically analyzed for EpCAM, proliferating cell nuclear antigen (PCNA) and co-localization of the two. EpCAM expression was mainly detected in hepatic tumor cells, showing a cytoplasmic staining pattern. However, expression was also slightly observed in normally-appearing surrounding hepatic cells. PCNA expression was highly detected in tumor cells, showing nuclear staining. Double staining of EpCAM and PCNA in tumors showed many cells with co-localization. Taken together, EpCAM and PCNA expression were increased in DEN-induced tumors and many tumor cells showed co-expression. It is suggested that EpCAM may increase during DEN-induced tumors, possibly associated with cell proliferation.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.407-414
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• 2011

The development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is dependent upon numerous factors. Central to development is the quality and developmental competence of the recipient cytoplast and the type of the donor nucleus. Typically metaphase of the second meiotic division (MII) has become the cytoplast of choice. Production of a cytoplast requires removal of the recipient genetic material, however, it may remove proteins which are essential for development or reduce the levels of cytoplasmic proteins to influence subsequent reprogramming of the donor nucleus. In this study, enucleation at MII did not affect the activities of either MPF or MAPK kinases. Immunocytochemical staining showed that both Cyclin B1 (MPF) and Erk1/2 (MAPK) were associated with the meiotic spindle of AI/TI oocytes with little staining in the cytoplasm, however, at MII association of both proteins with the spindle had reduced and a greater degree of cytoplasmic distribution was observed. The analysis of oocyte proteins removed during enucleation is a difficult approach to the identification of factors which may be depleted in the cytoplast. This is primarily due to the large numbers of aspirated karyoplasts which would be required for the analysis.

• 대한핵의학회:학술대회논문집
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• 대한핵의학회 2004년도 추계학술대회 및 총회
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• pp.408.2-408.2
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• 2004