• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.98-98
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• 2002

Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

• 환경위생공학
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• 제24권1호
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• pp.1-12
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• 2009

The various environmental problems that we face on today are basically about environmental quality. Since early 1960s affluent material was produced because of remarkable economic growth by many cooperations. However, for the lack of environmental policy, environmental pollutions has been serious. The central government should transfer the producing and consuming structure to environmental affinity through the regulations for developing Korea as a model of environmental nation which takes an active part in global environmental programme and in which the environment and economy are well harmonized. Moreover, the central government should take the lead in prevention of environmental pollution through the direct policies such as strengthening the discharge limit or setting up environmental basic institutions by securing budget for conserving environment. This thesis emphasize on the public institution among many environmental basic institutions for environmental anti-pollution project. It will find the problems with running those institutions, and will suggest the preview of improvement. Also, it is necessary to investigate of variation trend for inflow and pollutant loading to environmental infrastructure as increased of the diffusion rate as established and maintenance of sewer system. The purpose of this study is to investigate for inflow and pollutant loading to environmental infrastructure, and also to provide the method of efficiently maintenance and management. The results obtained were summarized as follows; 1. Survey of actual condition on operation and management of environmental infrastructure was evaluated the propriety of treatment process and problem of plant management. 2. Analysis of pollutant loading contribution for river system of environmental infrastructure with data analysis of water quality measuring network. 3. To investigate on case study for efficiently maintenance and management of environmental infrastructure. The result on this study was provide the method of efficiently maintenance and management with survey for establish and repair of sewer system and survey of actual condition on operation and management of environmental infrastructure in the water area of discharge to Geum River System. Application as guideline for establish and management of environmental infrastructure, and management of Geum River System. Also, application for preliminary data for fulfill-assess of total effluent regulation of water pollution.

• 대한소아치과학회지
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• 제28권3호
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• pp.447-463
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• 2001

Dendroaspis natriuretic peptide(DNP)는 Dendroaspis angusticeps의 뱀독으로부터 추출된 natriuretic peptide의 새로운 일원으로서, 사람의 혈장과 심방 근육에 존재하고 실험동물에서 혈관확장과 이뇨작용을 나타내는 것으로 알려져있다. 그러나 아직까지 심장 이외의 다른 기관내에 DNP가 존재하는지 또한 이들이 현재까지 알려진 기능 이 외의 또다른 생리적 기능을 나타내는 지의 여부는 아직 확실하게 밝혀져 있지 않다. 타액전 내에서 DNP의 기능을 규명하기 위한 전단계로, 먼저 흰쥐의 타액선내에 DNP가 존재하는지를 면역조직화학법을 이용하여 알아보았고 또한 DNP의 수용체가 타액전 또는 구강점막내에 존재하는 지의 여부를 생체외 방사선자기법을 이용하여 알아보았다. DNP의 면역반응은 악하선과 설하선 및 이하선에서 주로 도관부위에서 관찰되었고, 선포부위에서는 관찰되지 않았다. atrial natriuretic peptide(ANP)의 면역반응 역시 악하선과 설하선 및 이하선에서 DNP 면역반응과 유사하게 주로 도관부위에서 관찰되었다. DNP에 특이한 수용체가 타액선에는 저밀도로 존재한 반면 경구개와 혀 점막상피부위에서는 고밀도로 존재하였다. 경구개와 혀 점막상피부위의 DNP수용체는 주로 상피의 기저층과 기저층에 인접한 유극세포층에 국한되어 존재하는 것을 미세방사선자기법을 통하여 알 수 있었다. 이러한 결과로 볼 때 DNP가 타액선과 구강점막 부위에서 어떠한 생리적 기능을 나타낼 것으로 여겨지는데, 특히 구강점막 부위에서는 점막상피세포의 성장과 성숙과정에 관여하리라 추정된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3581-3586
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• 2014

Aim: To investigate the effects of tetramethypyrazine (TMP) on proliferation and apoptosis of the human gastric carcinoma cell line 7901 and its possible mechanism of action. Methods: The viability of TMP-treated 7901 cells was measured with a 3-(4, 5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT) and cell apoptosis was analyzed by flow cytometry. The distribution of cells in different phases of cell cycle after exposure of TMPs was analyzed with flow cytometry. To investigate the molecular mechanisms of TMP-mediated apoptosis, the expression of NF-${\kappa}Bp65$, cyclinD1 and p16 in SGC-7901 cells was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. Results: TMP inhibited the proliferation of human gastric carcinoma cell line 7901 in dose and time dependent manners. Cell growth was suppressed by TMP at different concentrations (0.25, 0.5, 1.0, 2.0 mg/ml), the inhibition rate is 0.46%, 4.36%, 14.8%, 76.1% (48h) and 15.5%, 18.5%, 41.2%, 89.8% (72h) respectively. When the concentration of TMPs was 2.0mg/ml, G1-phase arrest in the SGC-7901 cells was significant based on the data for cell cycle distribution. RT-PCR demonstrated that NF-${\kappa}Bp65$ and cyclin D1 mRNA expression was significantly down-regulated in 7901 cells treated with 2.0 mg/ml TMP for 72h (p<0.05), while the p16 mRNA level was up-regulated (p<0.05). The protein expression of NF-${\kappa}Bp65$ and cyclin D1 decreased gradually with the increase in TMP concentration, compared with control cells (p<0.05), while expression of protein p16 was up-regulated (p<0.01). Conclusion: TMP exhibits significant anti-proliferative and pro-apoptotic effects on the human gastric carcinoma cell line SGC-7901. NF-${\kappa}Bp65$, cyclinD1 and p16 may also play important roles in the regulation mechanisms.

• Asian-Australasian Journal of Animal Sciences
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• 제24권3호
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• pp.336-343
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• 2011

The neurotrophins, required for the survival and differentiation of the nervous system, are known to be important for the development of the reproductive tissues. However, the signals initiating the growth of follicles, gamete development, and transport and the development of zygote in the reproductive system of cows remain ambiguous. The purpose of the present study was to identify the transcripts and proteins of Neurotrophin 4 (NT4) and its receptor tyrosine kinase B (TrkB) in bovine reproductive tissues. The transcripts and immunoreactivity of NT4 and TrkB proteins were detected by reverse transcription polymerase chain reaction and western blot analysis. Using immunohistochemistry, the specific immunoreactivity of NT4 and TrkB were detected in the oocytes of primordial follicles and in the growing primary follicles. The NT4 and TrkB immunoreactivity was predominantly observed in granulosa cells, cumulus granulosa cells, cumulus oocyte complexes, theca cells of mature follicles, as well as in the oviduct epithelial cells, uterine gland cell, and epithelium cells of the uterus during the follicular and luteal phases in cows. Expressions of NT4 and TrkB mRNAs were not significantly different among the ovary, oviduct, and uterus of the follicular phase. For the luteal phase, the expression of NT4 mRNA in the ovary was significantly higher than that in the oviduct and uterus, and the expression of TrkB mRNA in the oviduct was significantly higher than that in the ovary and uterus, as determined by fluorescence quantitative reverse transcription polymerase chain reaction. The expression of NT4 mRNA was significantly higher than that of TrkB mRNA in the ovary and uterus, whereas NT4 mRNA expression was lower than that of TrkB mRNA in the oviduct during the luteal phase. The present study hypothesizes that NT4 participates in the regulation of both gonads and extra-gonadal reproductive tissues in cows.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5137-5142
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• 2012

UHRF2 is a member of the ubiquitin plant homeo domain RING finger family, which has been proven to be frequently up-regulated in colorectal cancer cells and play a role as an oncogene in breast cancer cells. However, the role of UHRF2 in glioma cells remains unclear. In this study, we performed real-time quantitative PCR on 32 pathologically confirmed glioma samples (grade I, 4 cases; grade II, 11 cases; grade III, 10 cases; and grade IV, 7 cases; according to the 2007 WHO classification system) and four glioma cell lines (A172, U251, U373, and U87). The expression of UHRF2 mRNA was significantly lower in the grade III and grade IV groups compared with the noncancerous brain tissue group, whereas its expression was high in A172, U251, and U373 glioma cell lines. An in vitro assay was performed to investigate the functions of UHRF2. Using a lentivirus-based RNA interference (RNAi) approach, we down-regulated UHRF2 expression in the U251 glioma cell line. This down-regulation led to the inhibition of cell proliferation, an increase in cell apoptosis, and a change of cell cycle distribution, in which S stage cells decreased and G2/M stage cells increased. Our results suggest that UHRF2 may be closely related to tumorigenesis and the development of gliomas.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5291-5298
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• 2012

Beta-asarone is one of the main bioactive constituents in traditional Chinese medicine Acorus calamu. Previous studies have shown that it has antifungal and anthelmintic activities. However, little is known about its anticancer effects. This study aimed to determine inhibitory effects on LoVo colon cancer cell proliferation and to clarify the underlying mechanisms in vitro and in vivo. Dose-response and time-course anti-proliferation effects were examined by MTT assay. Our results demonstrated that LoVo cell viability showed dose- and time-dependence on ${\beta}$-asarone. We further assessed anti-proliferation effects as ${\beta}$-asarone-induced apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide assay usinga flow cytometer and observed characteristic nuclear fragmentation and chromatin condensation of apoptosis by microscopy. Moreover, we found the apoptosis to be induced through the mitochondrial/caspase pathway by decreasing mitochondrial membrane potential (MMP) and reducing the Bcl-2-to-Bax ratio, in addition to activating the caspase-9 and caspase-3 cascades. Additionally, the apoptosis could be inhibited by a pan-caspase inhibitor, carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). When nude mice bearing LoVo tumor xenografts were treated with ${\beta}$-asarone, tumor volumes were reduced and terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assays of excised tissue also demonstrated apoptotic changes. Taken together, these findings for the first time provide evidence that ${\beta}$-asarone can suppress the growth of colon cancer and the induced apoptosis is possibly mediated through mitochondria/caspase pathways.

• 대한한의학방제학회지
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• 제14권1호
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• pp.105-119
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• 2006

Objectives & Methods : This present study was performed to investigate the effect of Hwadamtongrak-Tang extract (HTT) on the regulation of cerebral hemodynamics in terms of regional cerebral blood flow (rCBF) and mean arterial blood pressure (MABP)] in normal and cerebral ischemic rats. Also the effects of HTT on changes in local blood flow, inhibition of LD H activity in neuronal cells, and levels of cytokine production in the serum were determined in the ischemic rat model. The major findings are summarized below. Results : 1. HTT significantly increased rCBF in a dose-dependent manner, but MABP was not changed by HTT treatment. These results suggest that HTT may increase rCBF by dilating cerebral arterial diameter. 2. HTT-induced increase in rCBF was blocked by pretreatment with cyclooxygenase inhibitor indomethacin (IDN, 1 mg/kg, i.p.) and MABP was significantly increased by ID N. 3. Pretreatment of methylene blue $(MTB,\;10\;{\mu}g/kg,\;i.p.)$, an inhibitor of guanylate cyclase, significantly decreased both rCBP and MABP in HTT-treated rats. 4. HTT treatment significantly increased rCBP to a stable level during the period of cerebral reperfusion. 5. HTT significantly inhibited LD H activity in neuronal cells, suggesting a neuroprotection by HTT. 6. Serum interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ levels were significantly decreased in the femoral artery 1 hr after middle cerebral arterial occlusion in HTT-treated rats. IL-10 levels in the serum were significantly increased by HTT treatment whereas transforming growth factor $(TGF)-{\beta}$ levels were similar between HTT-treated and control groups. 7. Serum interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ levels were significantly decreased in the femoral artery 1 hr after reperfusion in HTT-treated rats. Serum IL-10 levels were significantly decreased in HTT-treated rats compared with the control group, and no significant changes in $(TGF)-{\beta}$ in the serum were observed by HTT treatment. Conclusions: The present data suggest that HTT may have an anti-ischemic effect via the improvement of cerebral hemodynamics and thus protect the brain from ischemic damage.

• 한국식품영양과학회지
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• 제41권7호
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• pp.907-913
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• 2012

포도, 대황, 사탕수수 등을 포함한 다양한 식물에서 발견되는 hydroxystilbene의 일종인 piceatannol은 암세포의 증식을 억제하고 apoptosis를 유발하는 것으로 알려져 있다. 본 연구에서는 AGS 인체위암세포를 대상으로 piceatannol에 의한 암세포 증식억제 과정에서 나타나는 또 다른 현상들을 조사하기 위하여 실시되었다. Piceatannol이 처리된 AGS 위암세포는 piceatannol의 처리 농도의 증가에 따라 생존율이 감소되었으며, 이는 세포주기 G1 arrest 유발과 연관이 있음을 MTT assay와 flow cytometry 분석을 통하여 확인하였다. Piceatannol에 의한 AGS 세포의 G1 arrest는 Cdks 및 cyclins의 발현 변화 및 Cdk 저해제인 p21의 발현을 전사 및 번역 수준에서 증가시켰으며, pRB 단백질의 인산화 감소 및 E2F1의 발현 억제와 연관성이 있었다. 아울러 piceatannol은 COX-2의 mRNA 및 단백질의 발현을 억제하였으나 COX-1의 발현에는 영향을 미치지 않았으며, piceatannol에 의한 COX-2의 발현억제는 PGE2의 생성 저하와 관련이 있었다. 본 연구의 결과는 piceatannol에 의한 세포주기 G1 arrest 유발이 COX-2의 선택적 발현 차단과 연관이 있음을 보여 주는 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제24권8호
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• pp.1128-1135
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• 2011

This study was conducted to investigate the effect of dietary green tea extract (GTE) on the performance, carcass and gastrointestinal tract (gut) traits, caecal coliform bacteria count, and pH and color (CIE $L^*$, $a^*$, and $b^*$) values of the breast muscle in broilers. A total number of 600 day-old broilers (Ross 308) was allocated to three treatments with four replicates containing 50 (25 males and 25 females) birds. The dietary treatments consisted of the basal diet as the control (0GTE) and diets with GTE at 0.1 (0.1GTE) or 0.2 (0.2GTE) g/kg. Body weights and the feed intake of broilers were measured at 1, 21 and 42 days, the feed intake was measured for different periods and the feed conversion ratio was calculated accordingly. At 42 day four birds per replicate were slaughtered for the determination of carcass and organ weights, caecal coliform bacteria count, and also quality of the breast muscle. The dietary GTE increased the body weight, feed efficiency, carcass weight and dressing percentage and decreased caecal coliform bacteria count of broilers (p<0.05). The 0GTE broilers consumed (p<0.01) less feed than the 0.1GTE birds in the entire experimental period. The relative abdominal fat weight and gut length of broilers in the 0.2GTE were tended to be lower (p<0.07) than those in the 0GTE group. The breast meat from 0.1GTE birds had a lower pH value when compared to that from 0GTE birds. The 0.1GTE broilers had lighter breast meat than 0GTE and 0.2GTE birds. The dietary GTE increased $a^*$ and $b^*$ values of the breast meat. Thus this product appeared to have a measurable impact on CIE color values of the breast meat in broilers. The results of the present study may indicate that the improved production results in the group with added GTE are directly connected with physiological mechanisms such as the regulation of the caecal micro-flora.