• Nutrition Research and Practice
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• 제9권2호
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• pp.129-136
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• 2015

BACKGROUND/OBJECTIVES: A variety of immunomodulators can improve the efficacy of low-dose chemotherapeutics. Active hexose correlated compound (AHCC), a mushroom mycelia extract, has been shown to be a strong immunomodulator. Whether AHCC could enhance the antitumor effect of low-dose 5-fluorouracil (5-FU) via regulation of host immunity is unknown. MATERIALS/METHODS: In the current study Hepatoma 22 (H22) tumor-bearing mice were treated with PBS, 5-FU ($10mg{\cdot}kg^{-1}{\cdot}d^{-1}$, i.p), or AHCC ($360mg{\cdot}kg^{-1}{\cdot}d^{-1}$, i.g) plus 5-FU, respectively, for 5 d. $CD^{3+}$, $CD^{4+}$, $CD^{8+}$, and NK in peripheral blood were detected by flow cytometry. ALT, AST, BUN, and Cr levels were measured by biochemical assay. IL-2 and $TNF{\alpha}$ in serum were measured using the RIA kit and apoptosis of tumor was detected by TUNEL staining. Bax, Bcl-2, and TS protein levels were measured by immunohistochemical staining and mRNA level was evaluated by RT-PCR. RESULTS: Diet consumption and body weight showed that AHCC had no apparent toxicity. AHCC could reverse liver injury and myelosuppression induced by 5-FU (P < 0.05). Compared to mice treated with 5-FU, mice treated with AHCC plus 5-FU had higher thymus index, percentages of $CD^{3+}$, $CD^{4+}$, and NK cells (P < 0.01), and ratio of $CD^{4+}$/$CD^{8+}$ (P < 0.01) in peripheral blood. Radioimmunoassay showed that mice treated with AHCC plus 5-FU had the highest serum levels of IL-2 and $TNF{\alpha}$ compared with the vehicle group and 5-FU group. More importantly, the combination of AHCC and 5-FU produced a more potent antitumor effect (P < 0.05) and caused more severe apoptosis in tumor tissue (P < 0.05) compared with the 5-FU group. In addition, the combination of AHCC and 5-FU further up-regulated the expression of Bcl-2 associated X protein (Bax) (P < 0.01), while it down-regulated the expression of B cell lymphoma 2 (Bcl-2) (P < 0.01). CONCLUSIONS: These results support the claim that AHCC might be beneficial for cancer patients receiving chemotherapy.

• Asian-Australasian Journal of Animal Sciences
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• 제25권5호
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• pp.621-628
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• 2012

The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2753-2758
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• 2014

Background: GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. Materials and Methods: GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Results: Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. Conclusions: These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.

• Journal of Nutrition and Health
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• 제24권4호
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• pp.356-365
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• 1991

유전에 의해 비만증을 타고나는 비만쥐 (ob/ob mouse)의 간으로 부터 유리된 세포막과 핵의 $L-triiodothyronine(T_3)$ 결합 능력이 그들과 한 배에서 태어난 정상 체중의 쥐들의 것들과 비교되었다. 세포막이 hypotonic 용액과 discontinuous sucrose density를 사용하여 원심분리기로 분리되었으며, 세포 각 부분의 marker enzyme들의 activity로 세포막의 순도가 측정되었다. 핵은 $Triton{\times}100$를 사용하여 윈심분리기로 얻어졌다. $T_3$ 수용체의 Bmax(최대결합용량)와 Kd(dissociation constant)가 세포막 혹은 핵을 여러 농도의 $^{125}I-T_3$와 함께 일정시간 incubation 시킨 후, 그 binding이 reverse Scatchard analysis에 의하여 계산하여 얻어졌다. 모든 실험과정은 비만쥐와 정상쥐에 대하여 평행으로 진행되었다. 간 핵의 $T_3$ 수용체의 최대결함용량은 비만쥐가 정상 체증의 쥐 보다 유의적으로 적었으나(p<0.001), $T_3$에 대한 친화력에는 차이가 없었다. 이는 이전의 보고들의 결과를 확인해 주는 것이다. 세포막에 있는 $T_3$ 수용체의 최대결합 능력과 친화력은 비만쥐와 정상쥐 간에 유의적인 차이가 없는 것으로 밝혀졌다. 이는 비만쥐의 세포막에 있는 $T_3$수용체의 기능에는 결함이 없음을 나타내며, 비만쥐의 말초조직에서 손상된 갑상선 홀몬의 작용은 세포막 수용체에 결합한 이후에 일어나는 과정에 원인이 있다는 것을 의미하고, 따라서 핵에 있는 $T_3$수용체의 결함이 비만쥐 (ob/ob mouse)의 비만증의 근본적인 원인일 수 있다는 제안을 뒷받침하여 주고 있다.

• Clinical and Experimental Reproductive Medicine
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• 제38권4호
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• pp.193-202
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• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• 한국환경과학회지
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• 제19권11호
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• pp.1423-1436
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• 2010

The effects of meteorological and reclaiming conditions on the reduction of suspended particles are investigated using a computational fluid dynamics (CFD) model with the k-$\varepsilon$ turbulence closure scheme based on the renormalization group (RNG) theory. Twelve numerical experiments with different meteorological and reclaiming conditions are performed. For identifying the meteorological characteristics of the target area and providing the inflow conditions of the CFD model, the observed data from the automatic weather station (AWS) near the target area is analyzed. Complicated flow patterns such as flow distortion, horse-shoe vortex, recirculation zone, and channeling flow appeared due to the topography and buildings in the domain. Specially, the flow characteristics around the reclamation area are affected by the reclaiming height, reclaiming size and windbreak height. Reclaiming height affected the wind speed above the reclaiming area. Windbreak induces more complicated flow patterns around the reclaiming area as well as within the reclaiming area. In front of the windbreak, flow is distorted as it impinges on the windbreak. As a result, upward flow is generated there. Behind the windbreak, a secondary circulation, so called, a recirculation zone is generated and flow is reattached at the end of the recirculation zone (reattachment point). At the lower part of the recirculation zone, there is a reverse flow toward the windbreak. Flow passing to the reattachment point starts to be recovered. Total amounts of suspended particles are calculated using the frictional and threshold frictional velocities, erosion potential function, and the number of surface disturbance. In the case of a 10 m-reclaiming and northerly wind, the amount of suspended particles is largest. In the presence of 5 m windbreak, the friction velocity above the reclaiming area is largely reduced. As a result, the total amount of the suspended particles largely decreases, compared to the case with the same reclaiming and meteorological conditions except for the windbreak The calculated suspended particle amounts are used as the emission rate of the dispersion model simulations and the dispersion characteristics of the suspended particles are analyzed.

• 한국지구물리탐사학회:학술대회논문집
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• 한국지구물리탐사학회 2004년도 정기총회 및 제6최 특별 심포지움
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• pp.25-48
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• 2004

구조물의 시간경과에 따른 변형 및 성능저하 현상은 인위적 요인 및 시간경과에 따른 지반상태의 변화 혹은 지반 변위요인의 누적에 의해 발생한다. 본 연구에서는 경상남도 경주시에 위치한 첨성대를 연구대상으로 설정하였다. 첨성대는 약 1,300여 년 전 건립된 석축구조물로서 지반침하로 약간 기울어져 있으며, 일부 석재 사이의 틈이 벌어져 있는 상태이다. 본질적인 대책수립을 위하여 본 연구에서는 이러한 목적을 위하여 3차원 레이저스캐닝시스템에 의한 (형상)역공학적 연구, 물리탐사에 의한 지반공학적 특성 연구 및 고유진동수 측정에 의한 첨성대의 동적특성 등의 다음과 같은 연구들을 수행하였다. 첫째, 정밀측량에 의한 첨성대의 정밀한 변형량 측정 및 지속적인 변위측정을 위하여 3차원 레이저 스캐닝에 의한 첨성대의 3차원 공간위치정보를 취득하여 형상역공학에 의한 3차원 벡터이미지 형상구현 및 첨성대의 크기, 변위량, 변위방향 등 여러 가지 제원을 구하고자 하였다. 둘째, 다양한 비파괴적 물리탐사 방법들을 적용하여 첨성대 및 주변 지반의 물성분포 및 지반특성을 파악하고자 하였다. 한편, 다양한 물리탐사를 통하여 향후 유사한 조사에 있어서 적절한 물리탐사 방법을 제시하고자 하였다. 셋째, 경주 첨성대의 1/10 모형에 대한 동적특성실험 및 실물에 대한 고유진동수 측정을 통해 첨성대의 구조적 특성을 연구하여 첨성대의 구조특성 및 상태판단 및 동적특성 파악에 의한 내진성능을 판단하고자 하였다.

• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.21-27
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• 1998

본 연구에서는 흰쥐 자궁에서 pituitary adenylate cyclase-activating polypeptide (PACAP)과 그 수용체 유전자들이 발현되는가와 각각 어떠한 유형의 transcript들이 발현되는가를 조사하였고, 이를 위해 역전사 중합효소 연쇄반응 (RT-PCR)을 시행하였다. 시상하부와 정소에서 공통적으로 존재함이 알려진 PACAP common exon 부위에 해당되는 primer를 사용하여 PCR을 시행한 결과 자궁을 포함한 모든 조직에서 에상대로 297 bp의 band가 확인되었다. 흰쥐 자궁에서 발현되는 PACAP mRNA에 정소 특이적인 exon 1이 포함되는가를 조사하기 위해 정소 특이적인 primer를 사용하여 PCR을 시행한 결과, 정소에서만 예상대로 586 bp의 band가 검출되었고 자궁을 포함한 다른 조직에서는 발견되지 않았다. 흰쥐의 PACAP 수용체 PCR에서는 자궁에서 923 bp와 839 bp크기의 band를 확인할 수 있었으며, 이는 알려진 PACAP type I수용체의 splicing variant 들 가운데 hip-hop (923bp)과 hip- 또는 nop-type (839bp)의 예상 크기와 일치하였다. 인위적으로 성적인 성숙을 유도한 PMSG 주사모델하에서 자궁내 PACAP transcript 수준은 주사 24 시간 후 실험군에서 증가하여 생식주기상 proestus 시기에 해당되는 주사 48 시간까지 증가하였다가 72 시간후에는 control 보다 낮은 수준을 보였는데, 이는 자궁의 PACAP 유전자발현이 생리적으로 조절됨을 나타내는 것이다. 본 연구는 흰쥐의 자궁에서 PACAP과 그 수용체 isoform들의 유전자가 발현됨을 최초로 보고한 것으로 자궁 자체에서 발현되는 PACAP이 autocrine 또는 paracrine하게 자궁의 생리 및 기능 조절을 담당함을 시사한다.

• 대한전자공학회논문지SD
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• 제38권10호
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• pp.678-686
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• 2001

CaAs metal semiconductor field effect transistor (MESFET) 소자의 전달컨덕턴스 분산 (transconductance dispersion) 현상과 게이트 누설 전류의 원인을 capacitance deep level transient spectroscopy (DLTS) 측정을 이용하여 해석하였다. DLTS 스펙트럼에서는 활성화 에너지가 각각 0.65×0.07 eV와 0.88 × 0.04 eV인 두개의 표면 결함과 0.84 × 0.01 eV의 활성화 에너지를 갖는 EL2를 관찰하였다. 전달컨덕턴스 분산 측정 결과, 전달컨덕턴스는 5.5 Hz ∼ 300 Hz의 주파수 영역에서 감소하였다. 전달컨덕턴스 분산을 온도의 함수로 측정한 결과, 온도가 증가할수록 전이 주파수는 증가하였고 전이 주파수의 온도 의존성으로부터 0.66 ∼ 0.02 eV의 활성화 에너지를 구할 수 있었다. 게이트 누설 전류 측정에서는 0.15 V 이하의 게이트 전압에서 순 방향과 역 방향 게이트 전압이 일치하는 오믹 전류-전압 특성을 나타내었고 게이트 누설 전류의 온도 의존성으로부터 구한 활성화 에너지는 0.63 ∼ 0.01 eV로 계산되었다. 서로 다른 방법으로 구한 활성화 에너지의 비교로부터 표면 결함 H1이 주파수에 따라서 감소하는 전달컨덕턴스 분산 및 게이트 누설 전류의 원인임을 알 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2087-2093
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• 2012

Previous evidence showed ${\beta}1$, 3-N-acetylglucosaminyltransferase 8 (${\beta}3GnT8$), which can extend polylactosamine on N-glycans, to be highly expressed in some cancer cell lines and tissues, indicating roles in tumorigenesis. However, so far, the function of ${\beta}3GnT8$ in laryngeal carcinoma has not been characterized. To test any contribution, Hep-2 cells were stably transfected with sense or interference vectors to establish cell lines that overexpressed or were deficient in ${\beta}3GnT8$. Here we showed that cell proliferation was increased in ${\beta}3GnT8$ overexpressed cells but decreased in ${\beta}3GnT8$ knockdown cells using MTT. Furthermore, we demonstrated that change in ${\beta}3GnT8$ expression had significant effects on tumor growth in nude mice.We further provided data suggesting that overexpression of ${\beta}3GnT8$ enhanced the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) at both the mRNA and protein levels, associated with shedding of tissue inhibitors of metalloproteinase TIMP-2. In addition, it caused increased production of transforming growth factor beta 1 (TGF-${\beta}1$), whereas ${\beta}3GnT8$ gene knockdown caused the reverse effect. The results may indicate a novel mechanism by which effects of ${\beta}3GnT8$ in regulating cellular proliferation are mediated, at least in partvia targeting MMPs/TIMPs and TGF-${\beta}1$ in laryngeal carcinoma Hep-2 cells. The finding may lay a foundation for further investigations into the ${\beta}3GnT8$ as a potential target for therapy of laryngeal carcinoma.