• Title/Summary/Keyword: 세포생존

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Effect of Sperm Preparation Technique on Subsequent In Vitro Development of Bovine Embryos (소 수정란 체외발달에 대한 정자분리방법의 효과)

  • 노규진
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.117-125
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    • 1998
  • 본 연구는 Percoll,Swim-up 그리고 Glass-wool 여과법의 세가지 정자분리 방법에 대한 정자 회수율, 생존율과 첨체반응율, 그리고 체외수정 후 배양시간에 따른 전핵형성율,수정란의 발달율과 세포할구수에 대한 효과를 조사하고자 실시하였다. 도축된 암소로 부터 채취한 난자를 22시간 체외배양 후 성숙된 난자를 체회수정시켰다. 수정 후 배양시간에 따라 존핵율을 조사하였으며 48시간에 분할율,192시간에 배반포기 발달율 및 세포할구수를 각각 비교 조사하였다. 정자의 첨체반응과 생존율은 처리군간에 차이가 없으나 회수율에 있어서 percoll 처리군이 다른 두 처리군보다 유의적으로 높았다(p<0.001).수정 후 배양시간별 전핵형성에 있어서도 percoll 처리군이 다른 두 처리군보다 빨리 진행됨을 볼 수 있다. 분할율에 있어서는 처리군간에 유의적 차이가 없으나.배반포기 발달율과 세포할구수에 있어서는 pecoll 처리군이 다른 두 처리군보다 유의적으로 높았다(P<0.05). 이상의 결과로 보아 percoll 처리에 대한 정자분리 방법은 정자 회수율이 높고 수정시 전핵형성 시간이 단축되어, 그 결과로 배반포기 발달율과 수정란의 세포할구수에 효과적임을 알 수 있었다.

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Overexpressed HRD3 Protein Required for Excision Repair of Schizosaccharomyces pombe is Toxic to the Host Cell (효모에서 절제회복에 관여하는 HRD3 유전자 과 발현이 숙주세포에 미치는 영향)

  • Choi In Soon
    • Environmental Analysis Health and Toxicology
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    • v.18 no.4
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    • pp.287-294
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    • 2003
  • 출아형 효모 Saccharomyces cerevisiae RAD3 유전자는 절제회복 및 세포의 생존에 필수적이며, DNA dependent ATPase와 DNA-RNA helicase활성을 가지고 있는 것으로 알려져 있다. 본 연구는 분열형 효모 Schizosaccharomyces pombe에서 절제회복과 세포의 생존에 필수적인 출아형 효모 RADS유전자와 유사한 유전자를 S. pombe genomic DNA library에서 분리하여 그 특성을 연구하였다. 분리한 RADS 유사유전자를 HRD3 유전자라 명명하였다. 발현 vector pET3a를 이용하여 분리한 HRD3 유전자를 과 발현하였을 때 HRD3단백질은 숙주단백질의 합성 억제 또는 분해 촉진을 유발하여 숙주세포인 대장균에 독성 효과를 나타냄이 관찰되었다. HRD3유전자와 lacZ유전자를 융합시킨 여러 가지 재조합 vector를 만들어 이들 융합단백질을 분리하였다. 이 결과 HRD3단백질의 카르복실 말단 부위가 DNA회복기능과 대장균에서의 독성효과를 나타내는 중요한 부위로 생각된다.

Treatment Results of Rhabdomyosarcomas of Head and Neck (두경부의 횡문근육종에서 다방면요법의 효과)

  • Ahn Ki-Jung;Suh Chang-Ok;Kim Gwi-Eon
    • Korean Journal of Head & Neck Oncology
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    • v.7 no.1
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    • pp.45-51
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    • 1991
  • 최근 횡문근육종을 치료함에 있어서 수술적 요법, 방사선 치료, 강암 약물요법을 적절히 병행함으로써 치료에 따르는 후유증을 극소화하면서 괄목할만한 생존율 향상을 가져오게 되었다. 특히 두경부의 횡문근육종은 소아에서 다발하고 병소의 위치에 따라서 각각 다른 임상적 특성을 가지며 수술적 절제를 했을 때 기능 및 외견상 결손이 크므로 별도의 연구 대상이 되어 왔으며 수술적 방법보다는 방사선 치료 및 항암 약물요법이 강조되고 있다. 저자들은 1976년부터 1987년까지 두경부 횡문근육종으로 진단받고 연세 암센터에서 항암 약물요법 및 방사선 치료를 받은 22명의 환자를 대상으로 하여 발생 부위, 조직 병리, 병기별 분포 및 생존율, 치료 방법에 따른 생존율을 후향성으로 분석하였다. 22명의 환자중 10 세 미만이 12 예로 가장 많았고, 13명이 배아세포형이였으며, 임상병기는 병기 III 이 가장 많아서 14 예였다. 병소의 위치는 안와가 6명으로 가장 많았고, 뇌수막주변부가 7명 (상악동 3명, 중이도 2명, 비강 l명, 비인강 1명), 기타 9명 (경부 5명, 이하선 2명, check 2명) 이었다. 5년 무병 생존율은 안와 종양에서 가장 높아서 50% 였고, 안와외 두경부가 37.5%, 뇌수막주변부가 16.7% 로 가장 낮았으며, 낮은 병기(병기 III), 배아세포형, 항암 약물 요법 병용군에서 상대적으로 높은 생존율이 관찰되었다.

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Effects of Baicalin on the Proliferation and Activity of Osteoblastic Cells (Baicalin이 조골세포의 생성 및 활성에 미치는 영향)

  • Ko, Seon-Yle
    • Journal of Oral Medicine and Pain
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    • v.33 no.2
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    • pp.105-110
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    • 2008
  • Baicalin is a flavonoid compound isolated from the medicinal plant Scutellaria baicalensis. It is known to affect multiple biological functions, including of antibacterial, anti-viral, anti-inflammatory and analgesic effects. Baicalin can inhibit nuclear factor-kappaB activation. It has been reported that some flavonoids possess the effects of bone metabolism. The present study was undertaken to determine the possible cellular mechanism of action of baicalin in osteoblasts. The effects on the osteoblast were determined by measuring cell proliferation, cell viability, alkaline phosphatase activity, and osteoprotegerin secretion. Baicalin has no effect on the osteoblastic cell proliferation and cell viability. Baicalin treatment showed increase in alkaline phosphatase activity and osteoprotegerin secretion of osteoblasts. Thus, baicalin may be a regulatory protein within the bone.

Transplantation of Marrow Stromal Cells into the Developing Mammal Retina (발생 중인 포유류 망막으로 골수기질세포의 이식)

  • Lee, Eun-Shil;Kwon, Oh-Ju;Ye, Eun-Ah;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.4
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    • pp.541-548
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    • 2013
  • Purpose: Marrow stromal cells (MSCs) have been known for their potential to trans-differentiate into neural and glial cells in vitro and in vivo. To investigate the influence of the developing host environment on the survival and morphological and molecular differentiation, murine MSCs transplanted into the eye of Brazilian opossum (Monodelphis domestica). Methods: Enhanced green fluorescent protein (GFP) - expressing MSCs were transplanted into developing Brazilian opossums. Animals were allowed to survive for up to 4 weeks after transplantation, at which time the eyes were prepared for immunohistochemical analysis. Results: Some transplanted MSCs survived and showed morphological differentiation into neural cells with some processes within the host vitreous chamber. Some transplanted cells expressed class III ${\beta}$-tubulin (TuJ1, a marker for neuronal cells) or glial fibrillary acid protein (GFAP, a marker for glial cells) or Nestin (a marker for neural stem cells). In addition, some transplanted cells were located in ganglion cell layer but did not show morphological and molecular differentiation. Conclusions: Our result show that the most effective stage of development for transplantation into the retina was postnatal day 16, which retinas developmentally corresponded to postnatal day 4-5 days mouse retina based on cell differentiation and lamination patterns. The present findings suggest that the age of the host appears to play a key role in determining cell fate in vivo.

Water Extract of Rhei Rhizoma Prevent Production of Reactive Oxygen Species and Loss of Mitochondrial Membrane Potential in a Hypoxia Model of Cultured Neurons (배양 신경세포의 저산소증모델에서 대황 물추출액의 항산화 및 사립체막전위 소실 억제 효능)

  • Lee, Hyun-Sook;Moon, Il-Soo
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1631-1636
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    • 2008
  • Rhei Rhizoma (RR; 大黃) consists of the underground parts (rhizome and root) of Rheum officinale Baill. and Rheum palmatum L. (Polygonaceae), and is widely used in Southeast Asian folk medicine to alleviate liver and kidney damages. In this study, we investigated into the efficacy and mechanism of RR water extract in supporting neuronal survival in a hypoxia model of cultured rat hippocampal neurons. RR exhibited no cytotoxicity up to 10 ${\mu}g$/ml and exhibited neurosupportive effects at 2.5 ${\mu}g$/ml in normoxia. When RR was added to the culture media on 10 days in vitro (DIV10) and given a hypoxic shock (2% $O_2$/5% $CO_2$, 3 hr, $37^{\circ}C$) on DIV13, RR exhibited neuroprotective effects on 5 days post-shock. $H_2DCF$ stainings indicated that RR effectively prevents ROS production in both normoxia and hypoxia. JC-1 stainings showed that RR prevents dissipation of MMP in hypoxia. These results indicate that RR protects neurons by suppressing ROS production and MMP loss.

Nodal Station as a Prognostic Factor in Resected Stage IIIA N2 Non-Small Cell Lung Cancer (절제된 IIIA N2 병기 비소세포형 폐암에 있어서 Nodal Station의 의의)

  • 김대준;김길동;김치영;정경영
    • Journal of Chest Surgery
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    • v.36 no.7
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    • pp.489-496
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    • 2003
  • To clarify the prognostic implication of the location and number of the metastatic mediastinal nodes in resected stage IIIA N2 non-small cell lung cancer. Material and Method: One hundred and seventy-four patients with resected non-small cell lung cancer who eventually proved to have pathologic stage IIIA N2 disease were studied. Patients who received preoperative induction therapy, non-curative operation or defined as operative mortality were excluded from this study. Result: In upper lobe tumors, there was no difference in 5-year survival according to the involvement of lower mediastinal nodes (32.3% vs 25.6%, p=0.86). In lower lobe tumors, no difference was found in 5-year survival according to the involvement of upper mediastinal nodes (25.1% vs 14.1%, p=0.33). There was no significant difference in 5-year survival between patients with or without metastatic subcarinal node (20.9% vs 25.6%, p=0.364). In terms of the number of metastatic mediastinal nodes, 5-year survival was better in single station group (26.3%) than multiple station group (18.3%) (p=0.048). In multiple station N2 group, the patients who received postoperative chemotherapy and radiation therapy had better 5-year survival (34.2%) (p=0.01). Cox's proportional hazards model revealed that the age $\geq$60 (O.R: 1.682, p=.006), multiple station N2 (O.R: 1.503. p=0.021), pneumonectomy (O.R: 1.562, p=0.018), postoperative chemotherapy and radiation therapy (O.R: 0.625, p=0.012) were the factors affecting the postoperative survival. Conclusion: Multiple station N2 disease was the important prognostic factor for postoperative survival in resected stage IIIA N2 non-small cell lung cancer. Postoperative chemotherapy and radiotherapy were thought to improve the survival in case of multiple station N2 disease.

Short-term Hypothermic Preservation of CHO Cells Using Serum-Free Media (무혈청 배지를 이용한 CHO 세포의 단기 저온보존)

  • Byoun, Soon-Hwi;Park, Hong-Woo;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.4
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    • pp.306-311
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    • 2006
  • Cell preservation is indispensable in animal cell culture process and should be established according to the cell characteristics. In this study, we experimented hypothermic preservation of CHO cells that is widely used in pharmaceutical industry to produce therapeutic proteins and established a stable method of preservation. The highest viability of CHO cells was obtained when the cells were preserved using rolling tube, which means the cells should be suspended to avoid the cell lumping during the preservation. Also, we obtained superior preservation result under the anaerobic condition. To evaluate the serum-free media as a preservation solution, we investigated cell growth after hypothermic preservation using serum-free media. High cell viability and normal cell growth was observed during 10 days using serum-free media. Moreover, we found that more effective preservation when ${\alpha}$-tocopherol and retinoic acid is added to media as an additive. In the case of 1 liter large scale hypothermic preservation using established protocol, cell viability and growth rate was obtained as good as small scale one. This study is considered to be helpful for hypothermic preservation of CHO cells and large scale hypothermic preservation may be available through the further studies.

Evaluation of Bisphenol a Induced Apoptosis in Sertoli Cell-lines (Bisphenol A에 의한 Sertoli 세포주 내 세포자연사 검정)

  • Kim, Ji-Hyang;Kim, Jin-Kyu;Do, Byoung-Rok;Lee, Chang-Joo;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.9 no.2
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    • pp.115-121
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    • 2005
  • The present report aimed at evaluating the effect of bisphenol A(BPA) and diethylstilbestrol(DES) on Leydig or Sertoli cell-lines. To identify the differences in the susceptibility to BPA upon different cell-types, assay of the cell viability was done on TM3(Leydig cells) and TM4(Sertoli cells) cell-lines. The result indicates that Sertoli cells are more sensitive to low dose of BPA than Leydig cells. Also, the BPA- or DES-treated Sertoli cells showed a reduction of phospholipase D(PLD) activity identically. According to the confirmation of the mRNA expression of fas receptor and fas ligand in the BPA-treated cells, fas/fasL system activated by BPA will deliver the apoptosis signal onto Sertoli TM4 cells. However, Fas/FasL system was not activated in the DES-treated cells unlike the BPA-treated cells.

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Correlation of Proliferating Cell Nuclear Antigen (PCNA) Expression and S-phase Fraction, Survival Rate in Primary Non-Small Cell Lung Cancer (원발성 비소세포 폐암에서 PCNA의 발현정도와 암세포의 분열능 및 생존률과의 관계)

  • Yang, Sei-Hoon;Kim, Hak-Ryul;Gu, Ki-Seon;Jung, Byung-Hak;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.756-765
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    • 1997
  • Background : To study the prognosis of patients with lung cancer, many investigators have reported the methods to detect cell proliferation in tissues including PCNA, thymidine autoradiography, flow cytometry and Ki-67. PCNA, also known as cyclin, is a cell related nuclear protein with 36KD intranuclear polypeptide that is maximally elevated in S phase of proliferating cells. In this study, PCNA was identified by paraffin-embedding tissue using immunohistochemistry which has an advantage of simplicity and maintenance of tissue architecture. The variation of PCNA expression is known to be related with proliferating fraction, histologic type, anatomic(TNM) stage, degree of cell differentiation, S-phase fraction and survival rate. We analyzed the correlation between PCNA expression and S-phase fraction, survival. Method : To investigate expression of PCNA in primary lung cancer, we used immunohistochemical stain to paraffin-embedded sections of 57 resected primary non-small cell lung cancer specimen and the results were analyzed according to the cell type, cell differentiation, TNM stage, S-phase fraction and survival. Results : PCNA expression was divided into five group according to degree of staging(-, +, ++, +++, ++++). Squamous cell type showed high positivity than in adenocarcinoma. Nonsignificant difference related to TNM stage was noticed. Nonsignificant difference related to degree of cell differentiation was noticed. S-phase fraction was increased with advance of PCNA positivity, but it could not reach the statistic significance. The 2 year survival rate and median survival time were -50% 13 months, +75% 41.3 months, ++73% 33.6 months, +++67% 29.0 months, ++++25% 9 months with statistic significance (P<0.05, Kaplan-Meier, generalized Wilcox). Conclusion : From this study, PCNA expression was high positive in squamous cell cancer. And, there was no relationship between PCNA positivity and TNM stage, cellular differentiation or S-phase fraction. But, the patients with high positive PCNA staining showed poor survival rate than the patients with lower positive PCNA staining (p<0.05). It was concluded that PCNA immunostaining is a simple and useful method for survival prediction in paraffin embedded tissue of non-small cell lung cancer.

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