• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3667-3671
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• 2015

Invasion and metastasis is the major cause of tumor recurrence, difficulty for cure and low survival rate. Excavating key transcription factors, which can regulate tumor invasion and metastasis, are crucial to the development of therapeutic strategies for cancers. PU.1 is a master hematopoietic transcription factor and a vital regulator in life. Here, we report that, compared to adjacent non-cancerous tissues, expression of PU.1 mRNA in metastatic hepatocellular carcinoma (HCC), but not primary HCC, was significantly down-regulated. In addition, levels of PU.1 mRNA in metastatic hepatoma cell lines MHCC97L and MHCC97H were much lower than in non-metastatic Hep3B cells. Transwell invasion assays after PU.1 siRNA transfection showed that the invasion of hepatoma cell lines was increased markedly by PU.1 knockdown. Oppositely, overexpression of PU.1 suppressed the invasion of these cells. However, knockdown and overexpression of PU.1 did not influence proliferation. Finally, we tried to explore the potential mechanism of PU.1 suppressing hepatoma cell invasion. ChIP-qPCR analysis showed that PU.1 exhibited a high binding capacity with miR-615-5p promoter sequence. Overexpression of PU.1 caused a dramatic increase of pri-, pre- and mature miR-615-5p, as well as a marked decrease of miR-615-5p target gene IGF2. These data indicate that PU.1 inhibits invasion of human HCC through promoting miR-615-5p and suppressing IGF2. These findings improve our understanding of PU.1 regulatory roles and provided a potential target for metastatic HCC diagnosis and therapy.

• 대한한의학방제학회지
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• 제28권3호
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• pp.255-269
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• 2020

Objectives : Hemistepta lyrata Bunge (Bunge) is a wild herb that has been used for managing fever and wound in Korean Traditional Medicine. The present study explored the effects of H. lyrata extract on liver X receptor (LXR) α-dependent lipogenic genes in hepatocyte-derived cells. Methods : After HepG2 cells or Huh7 cells were pre-treated with 1-10 ㎍/mL of H. lyrata extract or its fractionated extract for 0.5 h, the cells were subsequently exposed to LXR ligand for 6-24 h. Cell viability, LXR response element (LXRE)-driven luciferase activity, sterol regulatory element binding protein-response element (SREBP-RE)-driven luciferase activity, SREBP-1c expression, and mRNA levels of LXRα and its-dependent target genes were determined. In addition, LC-MS/MS analysis was conducted to explore major compounds in H. lyrata-chloroform fractionated extract #4 (HL-CF4). Results : Of various H. lyrata extracts tested, chloroform extract and its fractionated extract #4, HL-CF4, significantly decreased T0901317-mediated SREBP-1c expression. In addition, HL-CF4 significantly reduced LXRE atransactivation and LXRα mRNA expression without any cytotoxicity. Moreover, HL-CF4 prevented the SREBP-RE-driven luciferase activity and mRNA levels of fatty acid synthase and stearoyl-CoA desaturase-1 induced by T0901317. Results from LC-MS/MS analysis at positive/negative mode indicated that HL-CF4 contained several compounds showing m/z 197.1176 (C₁₁H₁₇O₃), 693.2913/227.1069 (C₃₈H₄₅O₁₂/C₁₅H₁₅O₂), 203.1797 (C₁₅H₂₃), 181.1225 (C₁₁H₁₇O₂), 591.2957 (C₃₅H₄₃O₈), 379.1040 (C₁₈H₁₉O₉), 409.1509 (C₂₀H₂₅O₉), 309.1348 (C₁₆H₂₁O₆), 391.1404 (C₂₀H₂₃O₈), and 669.2924/389.1248 (C₃₆H₄₅O₁₂/C₂₀H₂₁O₈). Conclusion : Based on its inhibition of the LXRα-dependent signaling pathway, H. lyrata chloroform extract and HL-CF4 have prophylactic potentials for managing non-alcoholic fatty liver.

• Pediatric Infection and Vaccine
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• 제3권2호
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• pp.154-162
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• 1996

Purpose : Adenoviruses(Ad) have been shown to play an important role in the etiology of severely acute respiratory diseases, particulary in infants and young children, and the occurrence of fatal outcome and chronic pulmonary sequelae in association with adenoviral infection has been a cause of great interest and concern. This report presents the resul of a retrospective analysis on 30 cases of lower respiratory infection from which adenovirus was isolated. Patients & Methods : The 30 patients in this study represent all detected cases of adenovial infection out of 240 children who were admitted to Sang Sung Medical Center between February to June 1996 showing signs and symptoms of lower respiratory tract infection. The diagnosis of adenovirus infection was based on microscopic visualization of typical cytopathic effect in HEp-2 tissue culture and used monoclonal Ab with nasopharyngeal aspiration. Results : The male/female ratio was 2:1 and the majority of age range was below 36months. Clinical diagnoses in all 30 patients were pneumonia(n=21), bronchitis and Bronchiolitis(n=5) and ARDS(n=4). We recieved the most of patients in the month of May. The chief complaints were fever(93.3%) and cough(80%) and extrapulmonary symptoms were diarrhea(n=5), seizure(n=4), abdominal pain(n=1). The mean duration of fever was $11.95{\pm}6.54$days. Physical examination on admission were crackles(73.3%), coarse breathing sounds(60%), hepatosplenomegaly(33.3%), decreased brething sounds(30%). In WBC counts, 8cases were below $4000/mm^3$ and 14 cases were above $10,000/mm^3$. In platelets counts, 4cases were below $150,000/mm^3$ and 10 cases were above $450,000/mm^3$. 21 cases were above 1 in CRP. GOT and GPT were abnormal in some cases. Chest X-ray revealed diffuse pulmonary infiltration(n=15), pleural effusion(n=6), consolidation(n=4) and hyperaeration(n=3). Seven patients were treated at the peditric intensive care unit with respiratory support and high dose of gammaglobulin. However, one patients died even through he was treated with NO ventilation and high frequency ventilation. Conclusion : Those with adenoviral pneumonia and respiratory infection having long fever duration and symptoms like bacterial pneumonia must be carefully differentiated in order to provide proper treatement and preventive measures due to possible fatal outcome.

• 대한한방내과학회지
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• 제26권1호
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• pp.93-106
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• 2005

Objectives : The aim of this study is to characterize the effect of Chungganhaeju-tang on $TGF-{\beta}l$-induced hepatic fibrosis. Materials and Methods : mRNA and protein expression levels of $TGF-{\beta}l$ in Chungganhaeju-tang treated HepG2 cells were compared to untreated cells using quantitative RT-PCR and ELISA assay, respectively. mRNA expression levels of the $TGF-{\beta}l$ signaling pathway genes $(T{\beta}R-I,\;T{\beta}R-II,\;Smad2,\;Smad3,\;Smad4,\;and\;PAI-1)$ and fibrosis-associated genes (CTGF, fibronectin, and collagen type $l{\alpha}$) were evaluated by quantitative RT-PCR. The effect of Chungganhaeju-tang on cell proliferation of T3891 human fibroblast was evaluated using $[^3H]Thymidine$ Incorporation Assay. Results : Inhibition of $TGF-{\beta}l$ mRNA expression and protein production was observed with treatment of Chungganhaeju-tang and seen to be dose and time dependent. Whereas $TGF-{\beta}l$-mediated induction of PAI-1 was suppressed with treatment of Chungganhaeju-tang, expression of the $TGF-{\beta}l$ signaling pathway genes such as $T{\beta}R-I$, $T{\beta}R-II$, Smad2, Smad3, and Smad4 was not affected. With treatment of Chungganhaeju-tang, inhibition of $TGF-{\beta}l$-induced cell proliferation of T3891 human fibroblast was observed, as well as abrogation of $TGF-{\beta}l$-mediated transcriptional up-regulation of CTGF, fibronectin, and collagen type $I{\alpha}$. Conclusion : This study strongly suggests that the liver cirrhosis-suppressive activity of Chungganhaeju-tang may be derived at least in part from its inhibitory effect on $TGF-{\beta}l$ functions, such as blockade of $TGF-{\beta}l$ stimulation of fibroblast cell proliferation and fibrosis-related gene expression as well as expression of $TGF-{\beta}l$ itself.

• Journal of Nutrition and Health
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• 제46권2호
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• pp.119-125
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• 2013

본 연구에서는 산수유, 목단피, 산약, 숙지황, 지골피, 야생배의 한약재 물 추출물이 당대사 관련 효소인 GCK, PDH, ACC mRNA 발현정도에 미치는 영향을 측정하였다. HepG2 세포에 대한 세포독성을 측정한 결과, GCK, PDH, ACC mRNA 발현량을 측정하기 위한 물 추출물의 농도 범위는 세포 생존율에 영향을 주지 않는 100, 250, 500 ppm로 결정하였다. GCK mRNA 발현은 100 ppm에서 숙지황 물 추출물이 165%로 가장 높게 나타났고, 250 ppm에서는 숙지황과 지골피 물 추출물이 각각 180%, 154%로 높았으며, 500 ppm에서는 산수유(195%), 목단피 (157%), 야생배 (139%), 산약 (122%), 지골피 (117%), 숙지황 (113%)의 순으로 GCK mRNA 발현이 증가되었다. PDH mRNA 발현량은 250 ppm 농도에서 지골피, 숙지황 물추출물에서 각각 141%, 118% 증가되었고, 500 ppm에서는 지골피, 숙지황 물 추출물에서 각각 191%, 124% 증가되었다. ACC mRNA 발현량은 500 ppm에서 지골피 (188%), 숙지황 (126%)로 가장 높게 나타났다. 결과적으로 GCK, PDH, ACC mRNA 발현량을 증가시킬 수 있는 소재로 산수유, 숙지황, 지골피 등을 꼽을 수 있겠으며 이들 소재들은 식후 혈당상승을 억제할 수 있는 항당뇨 천연소재로 이용될 수 있음을 제시하였다.

• 대한본초학회지
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• 제36권3호
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• pp.15-24
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• 2021

Objectives : The Glycyrrhiza new varieties, WONGAM and SINWONGAM, were developed through interspecific cross between Glycyrrhiza glabra and Glycyrrhiza uralensis by the National Institute of Horticultural and Herbal Science, Rural Development Administration in Korea. This in vitro study was undertaken to compare the antioxidant and cytotoxic effects between Glycyrrhiza new varieties (WONGAM and SINWONGAM) and official compendia (Glycyrrhiza glabra and Glycyrrhiza uralensis). Methods : Antioxidant activity was determined by DPPH (1,1-diphenyl-2-picrylhy drazyl), ABTS (2,2-azino-bis (3-rthylbenz-thiazoline-6-sulfonic acid)) diammonium salt, Nitrite radical scavenging assay, and Reducing Power assay. Cytotoxicity was determined by MTT assay and cell morphology was observed by an inverted microscope. Results : The DPPH, ABTS, Nitrite radical scavenging activities and reducing power of Glycyrrhiza glabra, Glycyrrhiza uralensis, WONGAM, and SINWONGAM were evaluated at different concentrations (0, 10, 50, 100, 500, 1000 ㎍/㎖). Glycyrrhiza glabra, Glycyrrhiza uralensis, WONGAM, and SINWONGAM showed similar dose-dependent increase in antioxidant activities. The cytotoxic effects with increasing doses of Glycyrrhiza new varieties and official compendia did not differ in HCT116, HT29, A549, MDA-MB231, PC3, ACHN, and HeLa cells. However, significant difference in cytotoxicity were observed in AGS, MCF7 and Hep3B cells by Glycyrrhiza glabra, Glycyrrhiza uralensis, WONGAM, and SINWONGAM. Conclusions : These results showed that Glycyrrhiza new varieties and official compendia acts as a potent antioxidant. Also, the finding that equivalent cytotoxic potency was observed in a cell dependent manner. Our study suggests that Glycyrrhiza new varieties may offer a wide-variety of health benefits.

• Nutrition Research and Practice
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• 제12권6호
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• pp.486-493
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• 2018

BACKGROUND/OBJECTIVE: The honeysuckle berry (HB) contains ascorbic acid and phenolic components, especially anthocyanins, flavonoids, and low-molecular-weight phenolic acids. In order to examine the potential of HB as a hepatoprotective medicinal food, we evaluated the in vitro anti-oxidant and anti-inflammatory activities of Korean HB (HBK) and Chinese HB (HBC). MATERIALS/METHODS: Antioxidant and anti-inflammatory effects of the extracts were examined in HepG2 and RAW 264.7 cells, respectively. The anti-oxidant capacity was determined by DPPH, SOD, CAT, and ARE luciferase activities. The production of nitric oxide (NO) as an inflammatory marker was also evaluated. The Nrf2-mediated mRNA levels of heme oxygenase-1 (HO-1), NAD(P)H dehydrogenase [quinone] 1 (Nqo1), and glutamate-cysteine ligase catalytic subunit (Gclc) were measured. The concentrations of HB extracts used were 3, 10, 30, 100, and $300{\mu}g/mL$. RESULTS: The radical scavenging activity of all HB extracts increased in a concentration-dependent manner (P < 0.01 or P < 0.05). SOD (P < 0.05) and CAT (P < 0.01) activities were increased by treatment with $300{\mu}g/mL$ of each HB extract, when compared to those in the control. NO production was observed in cells pretreated with 100 or $300{\mu}g/mL$ of HBC and HBK (P < 0.01). Treatment with $300{\mu}g/mL$ of HBC significantly increased Nqo1 (P < 0.01) and Gclc (P < 0.05) mRNA levels compared to those in the control. Treatment with $300{\mu}g/mL$ of HBK (P < 0.05) and HBC (P < 0.01) also significantly increased the HO-1 mRNA level compared to that in the control. CONCLUSIONS: Thus, the Korean and Chinese HBs were found to possess favorable in vitro anti-oxidant and anti-inflammatory activities. Nrf2 and its related anti-oxidant genes were associated with both anti-oxidant and anti-inflammatory activities in HB-treated cells. Further studies are needed to confirm these in vivo effects.

• 대한바이러스학회지
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• 제26권2호
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• pp.215-225
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• 1996

Aseptic meningits, an acute inflammation of the meninges, is a common illness during childhood. Virus is the most important cause of aseptic meningitis. Especially enterovirus causes approximately above 85% of all cases of aseptic meningitis. In 1993, there was a big epidemic of aseptic meningitis by ECHO 9 and ECHO 30 viruses. And ECHO 3 virus was isolated as a causative agent of aseptic meningitis in 1994. This study was aimed to detect the causative agent of aseptic meningitis in 1995 and to analyze the 5'-noncoding region which was used to detect virus. Virus was isolated from 87 stools and cerebrospinal fluid specimens of the patients by cultured RD and HEp-2 cell. Neutralizing antibody tests using enterovirus serum pool were performed on the specimens with cytopathic effect. 3 of ECHO 7 viruses and 5 of Coxsackie B3 viruses were isolated from stool specimens and 1 of ECHO 7 and Coxsackie B3 mixed type was confirmed from cerebrospinal fluid specimens. RNA was isolated from the culture supernatants of infected cells and general primers were selected in highly conserved part of the 5'-noncoding region of the enteroviral genome for RT-PCR. PCR product from this virus showed a 152bp band on gel electrophoresis. Sequence of obtained DNA was compared with prototype sequences by accessing to the Genebank database. 5'-noncoding region of isolated Coxsackie B3 virus, which has point mutations in nucleotide sequence positions 493, 497, 502, 523, was closely related to that of polio virus type 1, Mahoney strain. In case of isolated ECHO 7 virus, nucleotide has been changed from cytosine to thymine at position 581 and from thymine to cytosine at position 583. We concluded the causative agents of the outbreak of aseptic meningitis during June to July in 1995 were both ECHO 7 and Coxsackie B3 virus, and the primer used in this study could allow a rapid diagnosis of enteroviruses by PCR.

• BMB Reports
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• 제39권4호
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• pp.432-440
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• 2006

A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

• Molecular & Cellular Toxicology
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• 제5권4호
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• pp.298-303
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• 2009

(-)-Epigallocatechin-3-gallate (EGCG), a major flavonoid in green tea has multiple health benefits including chemoprevention, anti-inflammatory, anti-diabetic, and anti-obesity effects. In connection with these effects, EGCG can be a candidate to help the treatment of metabolic diseases. Metformin is a widely used anti-diabetic drug regulating cellular energy homeostasis via AMP-activated protein kinase (AMPK) activation. Therefore, the combination of metformin with EGCG may have additive or synergistic effects on treatment of type 2 diabetes. Nevertheless, there is no report for the pharmacokinetic and/or pharmacodynamic interaction of EGCG with metformin. Here, we evaluated the pharmacokinetic and pharmacodynamic interaction between metformin and EGCG in rats. Pharmacokinetics parameters of metformin were measured after oral administration of metformin in rats pre-treated with EGCG (10 mg/kg) or saline for 7 days. The results showed that there is no significant difference in pharmacokinetic parameters between saline control and EGCG-treated group. In addition, the hepatic AMPK activation by metformin in EGCG-treated rats was also similar to the control. The lack of additive effects of EGCG on AMPK activation or intracellular uptake of metformin was also evaluated in cells in the presence or absence of EGCG. Treatment of HepG2 cells with EGCG inhibited the metformin-induced AMPK activation. Combined results suggested that EGCG has no effect on the pharmacokinetics of metformin but may contribute to metformin action.