• 한국한의학연구원논문집
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• 제8권2호통권9호
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• pp.95-107
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• 2002

Yukmijihwang-tang(YM) is a noted herbal prescription in Chinese and Korean traditional medicines, and it has been known to reinforce the vital essence and has been widely used for a variety of disease such as stroke, osteoporosis, anti-tumor, and hypothyrodism. Regarding its traditional use, YM has been known to reinforce the Yin (vital essence) of liver and kidney. Also it has been known to reinforce nutrition and biological function in brain. Recently, studies suggested that YM increase antioxidant activities and exert the protective effect against oxidant-induced liver cell injury. We investigated the high-throughput gene expression analysis on the Yukmijihwang-tang administrated in SD rats. Microarray data were validated on a limited number of genes by semiquantitative RT-PCR and Western blot analyses. The recent availability of microarrays provides an attractive strategy for elaborating an unbiased molecular profile of large number of genes in drug discovery This experimental approach offers the potential to identify molecules or cellular pathways not previously associated with herbal medicine. Total RNA from normal control brain and Yukmijihwang-tang administrated brain were hybridized to microarrays containing 10,000 rat genes. The 52 genes were found to be up-regulated(twice or more) excluding EST gene. The nine genes were found to be down-regulated(twice or more) excluding EST gene. Gene array technology was used to identify for the first time many genes expression pathway analysis that arecell cycle pathway, apoptosis pathway, electron transport chain pathway, cytoplasmic ribosomal protein pathway, fatty acid degradation pathway, and TGF-beta signaling pathway. These differentially expressed genes pathway analysis have not previously been iavestigated in the context of herbal medicine efficacy and represent novel factors for further study of the mechanism of herbal medicine efficacy.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.1003-1012
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• 1996

The induction of a phenotype with preoperties may have clinical significance in the acceleration of the wound-healing process. Wound contraction involves a specialized cell known as the myofibroblast. The myofibroblasts can be identified by their intense staining of actin bundles with anti-actin antibody. Tissue-specific actin distribution is correlated with the contractile activity of the myofibroblasts and smooth muscle etc. This study was performed to determine the expression of actin filaments in the cytoplasm of cultured human gingival fibroblsts after GaAs laser(BIOSAER, Korea) irradiation. Human gingival fibroblasts were cultured from explants of normal interdental gingival tissue. The third-generation fibroblasts were used for immunohistochemical study. The cultured fibroblasts were exposed $0.53joule/cm^2$(lmW, 7 mimutes) of energy density, and then observed by immunohistochemical method using, rabbit anti0gelsolin, hen smooth muscle polyclonal antibody(Chemicon international inc.), and biotinylated goat anti-rabbit IgG(Vectastain) 24-, 36-, 48-hour after laser irradiation Following results were obtained ; 1. In nonirradiated cultures, round shaped active fibroblasts with abundant cytoplasm and prominet nucleoli were observed. 2. In 24- and 36-hour cultures after laser irradiation, spindle shaped cells with long process were observed. The intensity of stain was seen in cytoplasm of these modified fibroblasts. 3. In 48-hoour cultures after laser irradiation, stained spindle shape cell were not observed. The results suggest that the effect of the galium-arsenide laser treatment on cultured gingival fibroblasts is the rapid development of cytoplasmic actin filaments.

• BMB Reports
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• 제35권4호
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• pp.371-376
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• 2002

The receptor activator of nuclear factor kappa B (RANK) is a member of the tumor necrosis factor (TNF) receptor superfamily. It plays a critical role in osteoclast differentiaion, lymph node organogenesis, and mammary gland development. The stimulation of RANK causes the activation of transcription factors NF-${\kappa}B$ and activator protein 1 (AP1), and the mitogen activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK). In the signal transduction of RANK, the recruitment of the adaptor molecules, TNF receptor-associated factors (TRAFs), is and initial cytoplasmic event. Recently, the association of the MAPK kinase kinase, transforming growth factor-$\beta$-activated kinase 1 (TAK1), with TRAF6 was shown to mediate the IL-1 signaling to NF-${\kappa}B$ and JNK. We investigated whether or not TAK1 plays a role in RANK signaling. A dominant-negative form of TAK1 was discovered to abolish the RANK-induced activation of AP1 and JNK. The AP1 activation by TRAF2, TRAF5, and TRAF6 was also greatly suppressed by the dominant-negative TAK1. the inhibitory effect of the TAK1 mutant on RANK-and TRAF-induced NF-${\kappa}B$ activation was also observed, but less efficiently. Our findings indicate that TAK1 is involved in the MAPK cascade and NF-${\kappa}B$ pathway that is activated by RANK.

• BMB Reports
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• 제51권10호
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• pp.508-513
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• 2018

Fibronectin fragments found in the synovial fluid of patients with osteoarthritis (OA) induce the catabolic responses in cartilage. Nuclear high-mobility group protein Box 1 (HMGB1), a damage-associated molecular pattern, is responsible for the regulation of signaling pathways related to cell death and survival in response to various stimuli. In this study, we investigated whether changes induced by 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) in HMGB1 expression influences the pathogenesis of OA via an HMGB1-modulated autophagy signaling pathway. Human articular chondrocytes were enzymatically isolated from articular cartilage. The level of mRNA was measured by quantitative real-time PCR. The expression of proteins was examined by western blot analysis, immnunofluorescence assay, and enzyme-linked immunosorbent assay. Interaction of proteins was evaluated by immunoprecipitation. The HMGB1 level was significantly lower in human OA cartilage than in normal cartilage. Although 29-kDa FN-f significantly reduced the HMGB1 expression at the mRNA and protein levels 6 h after treatment, the cytoplasmic level of HMGB1 was increased in chondrocytes treated with 29-kDa FN-f, which significantly inhibited the interaction of HMGB1 with Beclin-1, increased the interaction of Bcl-2 with Beclin-1, and decreased the levels of Beclin-1 and phosphorylated Bcl-2. In addition, the level of microtubule-associated protein 1 light chain 3-II, an autophagy marker, was down-regulated in chondrocytes treated with 29-kDa FN-f, whereas the effect was antagonized by mTOR knockdown. Furthermore, prolonged treatment with 29-kDa FN-f significantly increased the release of HMGB1 into the culture medium. These results demonstrated that 29-kDa FN-f inhibits chondrocyte autophagy by modulating the HMGB1 signaling pathway.

• Natural Product Sciences
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• 제13권1호
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• pp.68-77
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• 2007

The aim of the present study was to examine the effects of green tea extract (CUMS6335) on the release of CA evoked by cholinergic stimulation and direct membrane-depolarization in the perfused model of the adrenal gland isolated from the spontaneously hypertensive rats (SHRs), and to establish the mechanism of action. Furthermore, it was also to test whether there is species difference between animals, and between CUMS6335 and EGCG, one of biologically the most powerful catechin compounds found in green tea. CUMS6335 $(100\;{\mu}g/ml)$, when perfused into an adrenal vein for 60 min, time-dependently inhibited the CA secretory responses evoked by ACh (5.32mM), high $K^+$(56 mM), DMPP $(100\;{\mu}M)$, and McN-A-343 $(100\;{\mu}M)$ from the isolated perfused adrenal glands of SHRs. However, CUMS6335 itself did fail to affect basal catecholamine output. Also, in adrenal glands loaded with CUMS6335 $(100\;{\mu}g/ml)$, the CA secretory responses evoked by Bay-K-8644 $(10\;{\mu}M)$ and cyclopiazonic acid $(10\;{\mu}M)$ were also inhibited in a relatively time-dependent fashion. However, in the Presence of EGCG $(8.0\;{\mu}g/ml)$ for 60 min, the CA secretory response evoked by ACh, high $K^+$, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were not affected except for last period. Collectively, these results indicate that CUMS6335 inhibits the CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by direct membrane-depolarization from the perfused adrenal gland of the SHR. It seems that this inhibitory effect of CUMS6335 is exerted by blocking both the calcium influx into the rat adrenal medullary chromaffin cells and the uptake of $Ca^{2+}$ into the cytoplasmic calcium store, which are at least partly relevant to the direct interaction with the nicotinic receptor itself. It seems likely that there is much difference in mode of the CA-releasing action between CUMS6335 and EGCG.

• 대한미생물학회지
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• 제12권1호
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• pp.33-49
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• 1977

These studies were carried out to detect the presence of mycotoxin fungi and detoxification of toxins in various kinds of grain and foodstuffs in Korea. The experiments were divided into three parts: mycologic, toxicologic and electron microscopic study. The resuIts were summarized as follows: 1. From the 210 various, samples(local grains, 150 samples; rice-cakes, 50 samples; meju, 10 samples), 1,205 colonies or fungi were isolated. In 1,127 of 1,205 colonies, it was possible to identify 21 genera. Among the identified strains, the predominant genera were Aspergillus sp.(28.5%), Penicillium sp.(27.1%) and Mucor sp.(7.8%). 2. In cytotoxicity test on HeLa cells, 25 strains showed severe toxic effects among the 240 strains of experiments. 3. In histopathologic test on mice, 21 strains showed severe toxic effect to mouse liver cells among the 240 tested strains. 4. In electron microscopic studies of HeLa cells and mouse liver cells from animal which had been treated with crude toxin, the liver cells showed the cytoplasmic change: dillatation or vacuolization of endoplasmic reticulum, swelling of mitochondria and disappearance of mitochondrial cristae, increased number of lipid and glycogen particles. Nucleus and nucelar envelope alterations, were also noted. 5. In the detoxification study with ultraviolet irradiation, 90% of the toxic substances were denatured by the ultraviolet light irradiation for 24 hours. 6. As a mass screening, the cytotoxicity test of HeLa cells and histopathologic study of mice liver cells treated with culture filtrates, employed and feasible to detect mycotoxin producing fungi.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• 제4권4호
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• pp.231-240
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• 2000

Effects of Jasmonic Acid and Wounding on Polyphenol Oxidase Activity in Senescing Tomato Leaves The effects of jasmonic acid(JA) and wounding on polyphenol oxidase(PPO) during leaf senescence was investigated by measuring the PPO activity in detached tomato(Lycopersicon esculentum Mill.) leaves of two-week-old seedlings. The PPO activity in the detached senescing leaves increased significantly in the dark. The leaf segments responded to the application of JA with accelerated senescence, as indicated by the loss of chlorophyll and rapid increase in the PPO activity. The senescence-promoting action of JA differed in the light and dark. Wounding the detached senescing leaves by scraping surface segments or making punctures with needles considerably delayed the loss of chlorophyll and had a significant effect on the PPO activity, the amounts of which were roughly proportional to the intensity of the wounding. In the dark, the combination of wounding plus JA resulted in stable levels of chlorophyll and PPO. JA and ABA acted similarly in both unwounded and wounded leaves, however, the amount of chlorophyll and PPO in the wounded segments was always higher than in the respective controls. JA was found to eliminate the senescence-retarding action of benzyladenine. In a histochemical localization test, the PPO activity was found to be localized in the cell walls of the parenchyma tissue, thereby indicating moderate cytoplasmic reactions. In the JA-treated plants, the PPO activity was intense in the cells of the cortex and phloem parenchyma. Accordingly, based on these observations it would appear that PPO is a component of a defense response maker, whereas JA plays an integral role in the intracellular signal transduction involved in inducible defense mechanisms.

• Asian Pacific Journal of Cancer Prevention
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• 제16권13호
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• pp.5359-5364
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• 2015

Background: Multiple complex pathways are operable in the evolution of cutaneous T cell lymphomas (CTCLs). These pathways involve interaction between neoplastic T cells and cells of the immune system (especially dendritic cells and the non-malignant T cells). Granulysin is a proinflammatory antimicrobial peptide which has an immune alarmin function, activating dendritic cells, as well as an active role in tumor immunology and prognosis. FOXP3+ regulatory T cells Tregs are an important player in the immune system. Much controversy is found in the literature about the role of Tregs in CTCL. Aim: The present study aimed to investigate the expression of granulysin and FOXP3 in mycosis fungoides (MF), its precursor lesion large plaque parapsoriasis and its leukemic form ;$s\acute{e}ezary$ syndrome (SS). Materials and Methods: Immunohistochemical expression of granulysin and FOXP3 were assessed in lesional skin biopsies taken from 58 patients (4 large plaque parapsoriasis, 48 MF and 6 SS). Results: Granulysin positivity was cytoplasmic and higher in MF than in parapsoriasis en plaque and higher in the more advanced stages of MF (p<0.001). All groups showed significant differences between each other except between MF tumor stage and SS. FOXP3 positivity was nuclear and higher in early stage MF (plaque and patch stages) than in tumor stages and SS (p<0.001). However the FOXP3 count was lower in parapsoriasis en plaque than in other stages of MF. All the groups showed significant differences between each other except between parapsoriasis and SS and between patch and plaque stages of MF. Conclusions: The present study supports a role for granulysin in MF progression and proposes a novel hypothesis about the effect of FOXP3 +veTregs in the suppression of the activity of the neoplastic cells in MF.

• 한국미생물·생명공학회지
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• 제33권3호
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• pp.184-188
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• 2005

HRF는 Na,K-ATPase의 ${\alpha}$ subunit에 결합하여 이의 활성을 저해하는 것으로 알려져 있으며, PKC에 의해 Ser98 잔기가 인산화 될 수 있다는 것을 anti-HRFpS98 항체와 HRF S98A mutant를 이용한 실험으로 확인할 수 있었다. 또한 $^{86}Rb^{+}-uptake$ assay 실험에서 HRF의 serine 98 잔기의 탈인산화는 Na,K-ATPase의 활성에 약간의 영향을 미치는 것으로 미루어 PKC에 의해 인산화되는 98 serine 잔기가 Na,K-ATPase 활성 저해에 큰 영향을 미치지 않는 것으로 보인다.

• 한국응용곤충학회지
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• 제39권3호
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• pp.193-197
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• 2000

Wolbachia는 몇몇 곤충류에 공생하는 세포내 공생세균으로서 수직 감염되며, 숙주의 세포질 불화합성, 단위생식, 자성화 등을 유도한다. 현재까지 알려진 Wolbachia는 비병원성이지만 돌연변이인 popcorn은 숙주인 초파리(Drosophila melanogaster)에 수직 감염되어 성충의 조직괴사와 조기 사망 등의 병원성을 나타낸다. Popcorn에 감염된 초파리는 알과 유충 발육기간이 연장되고, 번데기 기간은 단축되는 경향을 보이며, 성충의 수명이 단축되었다. 또한 비병원성 Wolbachia에 감염된 경우보다 산란율은 15%, 부화율은 80% 수준으로 감소하였고, 용화율은 67%, 우화율은 65%에 불과하였다. Popcorn과 비병원성 Wolbachia간의 교배에서 감염된 초파리는 모두 산란수의 감소와 차세대 부화율, 용화율, 우화율 등의 감소를 초래하였으며, popcorn에 감염된 초파리의 특성을 나타냈다. Wolbachia에 감염되지 않은 초파리와의 교배에서 popcorn 감염 초파리가 수컷인 경우 Wolbachia가 검출되지는 않았다. 따라서 popcorn은 초파리에서 모체 전염되지만, 세포질 분화합성은 유도하지 않으며, 초파리의 초기발육의 지연, 생식의 저하, 성충수명의 단축을 야기하는 등 병원성을 나타냈다.