• 대한한방종양학회지
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• 제6권1호
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• 대한치과교정학회지
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• 제26권1호
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• pp.65-82
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• 1996

본 연구는 II급 및 III급 부정교합자를 대상으로 activator사용에 따른 각 근육의 적응 및 변화양상을 알아보고자 시행되었다. 본 연구의 대상은 성장 발육기에 있는 아동중 II급 부정교합자 15명과 III급 부정교합자 17명으로 기능적 요인을 갖고있는 자로 선정하였고, 연구방법은 전측두근(T.A.), 교근(M.M.), 후측두근(T.P.), 악이복근 전복(D.A.)의 활성도를 알아보고자 $Biopak^{TM}$ program 과 쌍극표면전극을 이용하여 activator 치료 시작시기와 사용후 6-12개월 사이의 두차례에 걸쳐 안정위, 연하시, clenching등 세가지 조건에서 근활성도를 측정하여 비교해본 결과 다음과 같은 결론을 얻었다. II급 및 III급 부정교합자와 정상교합자의 근활성도를 비교한 결과 안정위시와 연하시 유의한 차이를 보이지 않았으나 clenching시 교근과 후측두근에서는 차이를 나타내었다. 치료전 근활성도가 정상보다 작은 군은 유의성 있는 증가를, 큰 군은 유의성 있는 감소를 보이는 등 대부분의 근육이 일정한 경향을 나타내었고, 이러한 경향은 III급 부정교합군에서 전측두근(연하시), 후측두근(clenching시), 악이복근 전복(연하시, clenching시)을 제외하고 모두에서 보였으며 II급 부정교합군에서는 전측두근(연하시), 교근(안정위, 연하시), 후측두근(clenching시)에서만 관찰되었다. 그러나 이러한 경향은 clenching시에서 감소되었다. II급 부정교합군과 III급 부정교합군과의 근활성도 비교에서 안정위시는 III급 부정교합군의 근활성도 증가율이 II급 부정교합군보다 우세했고, 연하시에는 교근을 제외하고 II급 부정교합군이, clenching시에는 역시 II급 부정교합군이 악이복근 전복을 제외하고 우세하게 나타났다.

• 한국경제지리학회지
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• 제21권1호
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• pp.20-33
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• 2018

도시공간에서 인구분포는 하루의 시간축을 따라 통행흐름의 변화와 함께 변동성을 보인다. 통행흐름은 도시민들이 하루 일과가 진행되면서 그들의 활동과 관련된 시설물이 입지한 지점으로 이동한 결과물이므로 도시민의 활동과 관련 시설물의 분포에 직접적인 영향을 받는다. 따라서 도시 인구분포의 시공간적 특성은 도시민의 일상생활과 관련된 활동공간의 분포와 그것을 방문하는 통행흐름을 결합하여 분석할 필요가 있다. 본 연구에서는 토지이용과 통행흐름에 대한 정보가 풍부한 서울시를 대상으로 건축물데이터베이스와 교통카드데이터베이스를 활용하여 하루 및 일주일 시간축을 따라 변동성을 보이는 도시공간의 인구분포를 분석한다. 일반적인 통계적 기법으로는 파악하기 어려운 시공간적 분석을 위하여 자바프로그램을 이용하여 시간과 공간을 결합한 4차원 시각화 방법을 고안한다. 이러한 4차원 시공간에서 역동적 시각화를 이용하면 직관적인 분석이 가능할 뿐만 아니라 인구분포의 시공간적 특성을 보다 효과적으로 파악할 수 있다. 이를 위하여 먼저 각 지점에 위치한 건축물을 용도에 따라 주거, 업무, 상업 활동으로 구분하고, 일주일분 교통카드데이터베이스에서 1분 단위로 각 지점의 버스와 지하철로 오가는 승객수를 산출하고, 이를 4차원으로 시각화하여 교통과 토지이용을 결합해서 서울시 인구분포의 시공간적 특성을 분석한다. 그 결과로 서울의 인구분포는 토지이용에 따라 뚜렷한 시공간적 특성을 보임을 파악할 수 있으며, 특히 업무활동, 상업활동, 주거활동의 혼합 양상에 따라 하루 시간축을 따라 인구분포 양상에 뚜렷한 차이가 있음을 확인하였다. 이러한 연구결과는 도시 시설의 입지계획과 교통계획 수립에 매우 유용하게 활용될 수 있다.

• Archives of Pharmacal Research
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• 제21권6호
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• pp.657-663
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• 1998

The enzyme responsible for the synthesis of nitric oxide (NO) from L-arginine in mammalian tissues is known as nitric oxide synthase (NOS) (EC.1.14.13.39). In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat pancreatic acinar cells. Treatment of rat pancreatic acinar cells with cholecystokinin-octapeptide (CCK-OP) resulted in an increase in the arginine conversion to citrulline, the amount of $NO_X$, the release of amylase, and the level of CGMP. Especially, CCK-OP-stimulated increase of arginine to citrulline transformation, the amount of $NO_X$, and CGMP level were completely counteracted by the inhibitor of NOS, NG-monomethyl-L-arginine (MMA), by contrast, that of amylase release was partially reduced. Furthermore, MMA-induced decrease of NOS activity and amylase release showed dose-dependent pattern. The data on the time course of CCK-OP-induced citrulline formation and CGMP rise indicate that NOS and guanylate cyclase were activated by treatment of CCK-OP. However, the mechanism of agonist-stimulated guanylate cyclase activation in acinar cells remains unknown. Therefore, activation of NOS is one of the early events in receptor-mediated cascade of reactions in pancreatic acinar cells and NO, not completely, but partially mediate pancreatic enzyme exocrine secretion.

• 대한한방내과학회지
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• 제29권2호
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• pp.401-412
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• 2008

Objectives : The aim of this study was to investigate the characteristics of Ryodoraku and association of Ryodoraku with gastric dysmotility in functional dyspepsia(FD). Methods : Subjects were 154 patients with FD and 18 patients with asthma. We calculated the average Ryodoraku score(RS, ${\mu}A$) and each variation from physiologic range of 12 Ryodoraku points, and investigated the incidence when left and right points were simultaneously below(bilateral deficiency) or above(bilateral excess) physiologic range. Postprandial regularity of normal slow waves, power ratio, and postprandial % of bowel sound were obtained by electrogastrography and enterotachography, and were used as gastric dysmotility index. Results : 1. Bilateral excess of H4, H5, F1, and F4 and bilateral deficiency of H4, H5, and H6 were characteristic in FD compared with asthma patients. 2. Incidence of gastric dysmotility in bilateral deficiency of H4, H5, and H6 was 100%, and was higher than in total FD patients(88.3%). 3. There was a positive correlation between the variation of H6 and % postprandial bowel sound. 4. Deficient tendency of H4, H5, and H6 was more evident when RS was above $40{\mu}A$ in FD. Conclusions : These findings suggest that gastric dysmotility in FD can be diagnosed when a pattern of H4, H5, H6 bilateral deficiency and F1, F4 bilateral excess is shown at the same time. We think this phenomenon is related to low activity of the vagus nerve rather than meridian pathway with result based on positive correlation between variation of H6 and postprandial % of bowel sound.

• BMB Reports
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• 제46권5호
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• pp.264-269
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• 2013

Pattern recognition receptors are known to participate in the activation of Prophenoloxidase system. In this study, a 1,3-${\beta}$-D-glucan recognition protein was detected for the first time in Antheraea pernyi larvae (Ap-${\beta}GRP$). Ap-${\beta}GRP$ was purified to 99.9% homogeneity from the hemolymph using traditional chromatographic methods. Ap-${\beta}GRP$ specifically bind 1,3-${\beta}$-D-glucan and yeast, but not E. coli or M. luteus. The 1,3-${\beta}$-D-glucan dependent phenoloxidase (PO) activity of the hemolymph inhibited by anti-Ap-${\beta}GRP$ antibody could be recovered by addition of purified Ap-${\beta}GRP$. These results demonstrate that Ap-${\beta}GRP$ acts as a biosensor of 1,3-${\beta}$-Dglucan to trigger the Prophenoloxidase system. A trace mount of 1,3-${\beta}$-D-glucan or Ap-${\beta}GRP$ alone was unable to trigger the proPO system, but they both did. Ap-${\beta}GRP$ was specifically degraded following the activation of proPO with 1,3-${\beta}$-Dglucan. These results indicate the variation in the amount of Ap-${\beta}GRP$ after specific immune challenge in A. pernyi hemolymph is an important regulation mechanism to immune response.

• Ocean and Polar Research
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• 제35권3호
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• pp.219-228
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• 2013

In Korea, more than 300 oil spill accidents occur every year. Despite the frequency, only a small pool of data is available on the initial effect of oil spill on macrobenthic fauna inhabiting rocky shores. The aim of this study was to analyze the variation of macrobenthic fauna composition and community structure on rocky shores, and understand the impact of oil on rocky shore organisms after the Hebei Spirit oil spill. Field surveys were carried out in five regions dose to the wreck site in January, April and September 2008. Polluted sites after the Hebei Spirit oil spill showed that biological index consistently decreased for 9 months limited to breeding and recruitment of organisms by spilled oil. Macrobenthic community was subdivided into 3 groups by species elimination and differences between density of major dominant species: enriched biota community under a relatively stable environment, the second with relatively low ecological index and the last with poor community. In this study, species number did not clearly reflect the effect of oil on the rare and mobile species. However, mean density, biomass and community structure showed the effect of oil by considering breeding activity, decline in recruitment and variation pattern with time.

• Molecules and Cells
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• 제41권4호
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• pp.342-350
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• 2018

Flowering time is determined by florigens. These genes include, Heading date 3a (Hd3a) and Rice FT 1 (RFT1) in rice, which are specifically expressed in the vascular tissues of leaves at the floral transition stage. To study the cis-regulatory elements present in the promoter region of Hd3a, we generated transgenic plants carrying the 1.75-kb promoter fragment of Hd3a that was fused to the ${\beta}$-glucuronidase (GUS) reporter gene. Plants expressing this construct conferred a vascular cell-specific expression pattern for the reporter gene. However, GUS was expressed in leaves at all developmental stages, including the early seedling stage when Hd3a was not detected. Furthermore, the reporter was expressed in roots at all stages. This suggests that the 1.75-kb region lackings cis-elements that regulate leaf-specific expression at the appropriate developmental stages. Deletion analyses of the promoter region indicated that regulatory elements determining vascular cell-specific expression are present in the 200-bp region between -245 bp and -45 bp from the transcription initiation site. By transforming the Hd3a-GUS construct to rice cultivar 'Taichung 65' which is defective in Ehd1, we observed that Ehd1 is the major regulatory element that controls Hd3a promoter activity.

• 한국수정란이식학회지
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• 제26권1호
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• pp.57-63
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• 2011

This study was performed to the expressions of pregnancy-associated plasma protein-A (PAPP-A) and 20alpha-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) in bovine corpus luteum during early pregnancy. To determine the function of PAPP-A gene during early pregnancy, we collected corpus luteum samples on 30, 60 and 90 days of pregnancy in bovine. The mRNA expression of PAPP-A, $20{\alpha}$-HSD, progesterone-receptor (PR) and insulin-like growth factor binding protein4 (IGFBP4) gene was conducted by Real-time PCR. In parallel with mRNA levels, The protein expressions of PAPP-A and $20{\alpha}$-HSD were detected by immunological analysis. The mRNA expressions $20{\alpha}$-HSD and PAPP-A significantly increased on day 90 in the corpus luteum during pregnancy. The mRNA expression of PR and JGFBP4 in the corpus luteum progressively was enhanced at 30 to 60 day, but decreased on 90 day of pregnancy in the corpus luteum. The expression patterns of these genes, PAPP-A and $20{\alpha}$-HSD were similar pattern in these tissues. In conclusion, PAPP-A and $20{\alpha}$-HSD activity in corpus luteum could be played a role for early pregnancy manifestation.

• 한국수정란이식학회지
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• 제33권3호
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• pp.99-105
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• 2018

To determine the differences in the in-vitro ovum maturation process of bovine, we compared the expression of MMPs in these oocytes and cumulus cell throughout oocytes maturated. In an attempt to investigate the effect of MMP activation and inhibitors in total protein of cumulus cell and, oocytes during oocytes maturation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), TIMPs (TIMP-2 and TIMP-3), as well as their expression profiles (Real-time PCR, Gelatin Zymography and ELISA). Our results that the bovine oocytes MMP-2 and MMP-9 level was significantly associated with the rate of maturity of oocytes (P<0.05). In cumulus cell, MMP-2 was highly expressed in all stages of the oocyte's maturation. The final oocytes maturation exhibited strong gelatinase activity. There was no significant correlation between cumulus cell MMP-9 and the maturation rate of oocytes. However, for the oocyte cytoplasm MMP-9 expression was significant correlation to the maturation oocytes. There was no significant correlation between cumulonimbus cells MMP-9 and oocyte maturation rates; however, for oocyte cytoplasm, MMP-9 expression was significantly correlated with mature oocyte. However, the TIMP-1 and TIMP-2 protein expression patterns are not correlated with the maturation rate of the oocyte. Our results suggest that MMP different expression pattern may regulate the morphological remodeling of oocyte's in the cumulus cell. Further, the MMP-2 expression has a strong relation with a higher maturation rate of the oocyte.