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Inhibitory Effect of Amentoflavone of Selaginella Tamariscina on MMP-9 Expression through NF-${\kappa}$B and AP-1 in Macrophage Raw 264.7 cells

  • Ahn, Byung-Tae;Shin, Sung-Ahn;Kim, Jun-Gi;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.243-249
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    • 2007
  • The French paradox has been attributed to the antioxidant properties of flavonoids present in the red wine. Amentoflavone(AF) is a bi-flavonoid compound with anti-fungal and anti-inflammatory activities. We isolated AF from Selaginella tamariscina, and studied its effects on nuclear factor-B(NF-B)-mediated MMP-9 gene expression in RAW264.7 cells. AF blocked the lipopolysaccharide(LPS)-induced expression of MMP-9. Zymographic and immunoblot analyses showed that AF suppressed LPS-induced MMP-9 expression in a dose-dependent manner. To clarify the mechanistic basis for its inhibition of MMP-9 induction, we examined the effect of AF on the transactivation of MMP-9 gene by luciferase reporter activity using -1.59 kb flanking region. AF potently suppressed the reporter gene activity. This inhibition was characterized by down-regulation of MMP-9, which was transcriptionally regulated at NF-B site and activation protein-1 (AP-1) site in the MMP-9 promoter, two important nuclear transcription factors that are involved in MMP-9 expression. These findings indicate the efficacy of AF in inhibiting MMP-9 expression through the transcription factors NF-B and AP-1 on LPS-induced RAW264.7 cells.

IGF1 potentiates BMP9-induced osteogenic differentiation in mesenchymal stem cells through the enhancement of BMP/Smad signaling

  • Chen, Liang;Zou, Xiang;Zhang, Ran-Xi;Pi, Chang-Jun;Wu, Nian;Yin, Liang-Jun;Deng, Zhong-Liang
    • BMB Reports
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    • v.49 no.2
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    • pp.122-127
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    • 2016
  • Engineered bone tissue is thought to be the ideal alternative for bone grafts in the treatment of related bone diseases. BMP9 has been demonstrated as one of the most osteogenic factors, and enhancement of BMP9-induced osteogenesis will greatly accelerate the development of bone tissue engineering. Here, we investigated the effect of insulin-like growth factor 1 (IGF1) on BMP9-induced osteogenic differentiation, and unveiled a possible molecular mechanism underling this process. We found that IGF1 and BMP9 are both detectable in mesenchymal stem cells (MSCs). Exogenous expression of IGF1 potentiates BMP9-induced alkaline phosphatase (ALP), matrix mineralization, and ectopic bone formation. Similarly, IGF1 enhances BMP9-induced endochondral ossification. Mechanistically, we found that IGF1 increases BMP9-induced activation of BMP/Smad signaling in MSCs. Our findings demonstrate that IGF1 can enhance BMP9-induced osteogenic differentiation in MSCs, and that this effect may be mediated by the enhancement of the BMP/Smad signaling transduction triggered by BMP9.

Dietary fat preference and effects on performance of piglets at weaning

  • Weng, Ruey-Chee
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.834-842
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    • 2017
  • Objective: An experiment was to evaluate the interplay of dietary lipid sources and feeding regime in the transition from sow milk to solid food of abruptly weaned piglets. Methods: Soon after weaning, 144 piglets were selected and were trained over a 15 day period to experience gradually reducing dietary fat content from 12% to 6% for lard (L), soybean oil (S), and coconut oil (C) and their feeding behavior and diet preference then tested in a behavior observation experiment. Another 324 weaned piglets were used in three consecutive feeding experiments to measure the effect of different dietary fats on performance and feed choice in the four weeks after abrupt weaning. The lipid sources were used as supplements in a 3% crude fat corn/soya basal diet, with 6% of each being included to form diets 9C, 9S, and 9L respectively, and their effects on performance measured. Combinations of these diets were then further compared in fixed blends or free choice selection experiments. Results: Piglets pre-trained to experience reducing lipid inclusion showed different subsequent preferences according to lipid source, with a preference for lard at 9%, soybean oil at 3%, and coconut oil at 6% inclusion rate (p<0.001). Following abrupt weaning, whilst after 4 weeks those fed 9C had the heaviest body weights (18.13 kg, p = 0.006). Piglets fed a fixed 1:1 blend of 9C+9S had a poorer feed conversion ratio (FCR = 1.80) than those fed a blend of 9C+9L (FCR = 1.4). The 9C and 9L combination groups showed better performance in both fixed blend and free choice feeding regimes. Conclusion: After abrupt weaning, they still have dependence on high oleic acid lipids as found in sow milk. A feeding regime offering free choice combination of lipids might give the possibility for piglets to cope better with the transition at weaning, but further research is needed.

Correlation of expression and activity of matrix metalloproteinase-9 and -2 in human gingival cells of periodontitis patients

  • Kim, Kyung-A;Chung, Soo-Bong;Hawng, Eun-Young;Noh, Seung-Hyun;Song, Kwon-Ho;Kim, Hanna-Hyun;Kim, Cheorl-Ho;Park, Young-Guk
    • Journal of Periodontal and Implant Science
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    • v.43 no.1
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    • pp.24-29
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    • 2013
  • Purpose: Matrix metalloproteinases (MMPs) are capable of degrading extracellular matrix, and they are inducible enzymes depending on an inflammatory environment such as periodontitis and bacterial infection in periodontal tissue. Gingival inflammation has been postulated to be correlated with the production of MMP-2 and MMP-9. The objective of this study was to quantify the expression and activity of MMP-9 and -2, and to determine the correlation between activity and expression of these MMPs in human gingival tissues with periodontitis. Methods: The gingival tissues of 13 patients were homogenized in $500{\mu}L$ of phosphate buffered saline with a protease inhibitor cocktail. The expression and activity of MMP-2 and -9 were measured by enzyme-linked immunosorbent assay and Western blot analysis, and quantified by a densitometer. For the correlation line, statistical analysis was performed using the Systat software package. Results: MMP-9 was highly expressed in all gingival tissue samples, whereas MMP-2 was underexpressed compared with MMP-9. MMP-9 activity increased together with the MMP-9 expression level, with a positive correlation (r=0.793, P=0.01). The correlation was not observed in MMP-2. Conclusions: The expression of MMP-2 and -9 might contribute to periodontal physiological and pathological processes, and the degree of MMP-9 expression and activity are predictive indicators relevant to the progression of periodontitis.

Electrochemical Immunosensor Based on the ZnO Nanorods Inside PDMS Channel for H7N9 Influenza Virus Detection (PDMS 채널 내부에 성장된 산화아연 나노막대를 이용한 H7N9 인플루엔자 바이러스 전기화학 면역센서)

  • Han, Ji-Hoon;Lee, Dongyoung;Pak, James Jungho
    • Journal of Sensor Science and Technology
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    • v.23 no.4
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    • pp.278-283
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    • 2014
  • In this study, we propose an immunosensor using zinc oxide nanorods (NRs) inside PDMS channel for detecting the influenza A virus subtype H7N9. ZnO with high isoelectric point (IEP, ~9.5) makes it suitable for immobilizing proteins with low IEP. In this proposed H7N9 immunosensor structure ZnO NRs were grown on the PDMS channel inner surface to immobilize H7N9 capture antibody. A sandwich enzyme-linked immunosorbent assay (ELISA) method with was used 3,3',5,5' tetramethylbenzidine (TMB) for detecting H7N9 influenza virus. The immunosensor was evaluated by amperometry at various H7N9 influenza antigen concentrations (1 pg/ml - 1 ng/ml). The redox peak voltage and current were measured by amperometry with ZnO NWs and without ZnO NWs inside PDMS channel. The measurement results of the H7N9 immunosensor showed that oxidation peak current of TMB at 0.25 V logarithmically increased from 2.3 to 3.8 uA as the H7N9 influenza antigen concentration changed from 1 pg/ml to 1 ng/ml. And then we demonstrated that ZnO NRs inside PDMS channel can improve the sensitivity of immunosensor to compare non-ZnO NRs inside PDMS channel.

Effects of $(1R,9S)-{\beta}-Hydrastine$ hydrochloride on L-DOPA-Induced Cytotoxicity in PC12 cells

  • Yin, Shou-Yu;Lee, Jae-Joon;Kim, Yu-Mi;Jin, Chun-Mei;Yang, Yoo-Jung;Lee, Myung-Koo
    • Natural Product Sciences
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    • v.10 no.3
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    • pp.124-128
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    • 2004
  • Previously, $(1R,9S)-{\beta}-Hydrastine$ hydrochloride has been found to lower dopamine content in PC12 cells (Kim et al., 20001). In this study, the effects of $(1R,9S)-{\beta}-Hydrastine$ hydrochloride on L-DOPA-induced cytotoxicity in PC12 cells were investigated. Treatment with $(1R,9S)-{\beta}-Hydrastine$ hydrochloride at concentrations higher than $500\;{\mu}M$ caused cytotoxicity in PC12 cells. In addition, $(1R,9S)-{\beta}-Hydrastine$ hydrochloride at non-cytotoxic or cytotoxic concentrations significantly enhanced L-DOPA-induced cytotoxicity (L-DOPA concentration, $50\;{\mu}M$). Treatment of PC12 cells with $750\;{\mu}M$ $-1R,9S)-{\beta}-Hydrastine$ hydrochloride and $50\;{\mu}M$ L-DOPA, alone or in combination, also induced cell death via a mechanism which exhibited morphological and biochemical characteristics of apoptosis, including chromatin condensation and membrane blebbing. Exposure of PC12 cells to $(1R,9S)-{\beta}-Hydrastine$ hydrochloride, L-DOPA and $(1R,9S)-{\beta}-Hydrastine$ hydrochloride plus L-DOPA for 48 h resulted in a marked increase in the cell loss and percentage of apoptotic cells compared with exposure for 24 h. These data indicate that $(1R,9S)-{\beta}-Hydrastine$hydrochloride at higher concentration ranges aggravates L-DOPA-induced neurotoxicity cytotoxicity in PC12 cells. Therefore, it is proposed that the long-term L-DOPA therapeutic patients with $(1R,9S)-{\beta}-Hydrastine$ hydrochloride could be checked for the adverse symptoms.

Effect of Allergen Removed Rhus Verniciflua Extract on Inhibition of Tumor Metastasis (알러젠 제거 옻나무 추출물이 종양 전이 억제에 미치는 영향)

  • Park, Jae-Hyun;Moon, Goo
    • Journal of Korean Traditional Oncology
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    • v.15 no.1
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    • pp.47-61
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    • 2010
  • Objective : The aim of this present study is to evaluate the inhibitory effect of allergen removed Rhus verniciflua (ARV) on Matrix Metalloproteinase-9 (MMP-9), Matrix Metalloproteinase-2 (MMP-2) which is considered to have a clinically important role in tumor metastasis. Methods : The inhibitory effects of standardized extract of ARV on the MMP-2, MMP-9 were investigated by spectrofluorometer while the inhibitory effects on the active MMP-2, pro MMP-2, pro MMP-9 were investigated by zymography. Antimetastatic effect of standardized extract of ARV was investigated in vitro on human fibrosarcoma cell (HT1080)'s invasion through Matrigel. Results : The standardized extract of ARV showed inhibitory effects on the active MMP-2 (IC50, $1.01{\mu}g$/ml), active MMP-9 (IC50, $2.5{\mu}g$/ml) depending on concentrations which was determined by spectrofluorometer. The standardized extract of ARV showed inhibitory effects on the active MMP-2, pro MMP-2, pro MMP-9 depending on concentrations which was determined by zymography. However its inhibitory effect on pro MMP-9 was relatively weaker rather than active MMP-2, pro MMP-2. The standardized extract of ARV showed inhibitory effects in vitro on human fibrosarcoma cell (HT1080)'s invasion through Matrigel according to concentration. Conclusions : These results indicate that standardized extract of ARV has antimetastatic effect through inhibit again MMP-2, MMP-9. Also its inhibitory effect is more powerful on active MMP-2, pro MMP-2 than on active MMP-9, pro MMP-9. It is necessary to conduct further studies on other MMP families, TIMP, and each component of standardized extract of ARV.

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microRNA-214-mediated UBC9 expression in glioma

  • Zhao, Zhiqiang;Tan, Xiaochao;Zhao, Ani;Zhu, Liyuan;Yin, Bin;Yuan, Jiangang;Qiang, Boqin;Peng, Xiaozhong
    • BMB Reports
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    • v.45 no.11
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    • pp.641-646
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    • 2012
  • It has been reported that ubiquitin-conjugating enzyme 9 (Ubc9), the unique enzyme2 in the sumoylation pathway, is up-regulated in many cancers. However, the expression and regulation of UBC9 in glioma remains unknown. In this study, we found that Ubc9 was up-regulated in glioma tissues and cell lines compared to a normal control. UBC9 knockdown by small interfering RNA (siRNA) affected cell proliferation and apoptosis in T98G cells. Further experiments revealed that microRNA (miR)-214 directly targeted the 3' untranslated region (UTR) of UBC9 and that there was an inverse relationship between the expression levels of miR-214 and UBC9 protein in glioma tissues and cells. miR-214 overexpression suppressed the endogenous UBC9 protein and affected T98G cell proliferation. These findings suggest that miR-214 reduction facilitates UBC9 expression and is involved in the regulation of glioma cell proliferation.

Normalization of CA19-9 Following Resection for Pancreatic Ductal Adenocarcinoma is not Tantamount to being Cured?

  • Chen, Tao;Zhang, Min-Gui;Yu, Xian-Jun;Liu, Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.661-666
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    • 2015
  • Background: Postoperative carbohydrate antigen 19-9 (CA19-9) is an independent predictor of survival for pancreatic ductal adenocarcinoma (PDAC), and more powerful than preoperative CA19-9. However, making decisions just dependent on postoperative CA19-9 may result in necessary treatments not being performed. Materials and Methods: A total of 178 patients with resected PDAC were eligible for this retrospective study, classified into two corresponding subgroups according to postoperative CA19-9. Prognostic significance of all clinicopathologic factors was evaluated by univariate and multivariate analyses. Results: Postoperative CA19-9, preoperative CA125 and lymph node status were independent predictors. Better predictive performances for overall survival (OS) and recurrence-free survival (RFS) were achieved by postoperative CA19-9 compared to preoperative CA125 and lymph node status. Particularly, preoperative CA125 was associated with poor OS (p<0.001 for the normalized CA19-9 patients, p=0.012 for the elevated) and RFS (p=0.005 for the normalized, p=0.004 for the elevated). Moreover, preoperative CA125 levels related with survival in double-negative patients. Conclusions: Normalization of CA19-9 is not tantamount to be cured. Preoperative CA125 is a critical predictor for PDAC patients, especially in double-negative patients.

Association of Matrix Metalloproteinase (MMP)-2 and -9 Expression with Extra-gastrointestinal Stromal Tumor Metastasis

  • Wang, Chao;Ma, Hong-Xi;Jin, Mei-Shan;Zou, Ya-Bin;Teng, Yong-Liang;Tian, Zhuang;Wang, Hai-Ying;Wang, Yin-Ping;Duan, Xiu-Mei
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4187-4192
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    • 2014
  • Matrix metalloproteinase (MMP)-2 and MMP-9 are important proteases involved in invasion and metastasis of various tumors. Extra-gastrointestinal stromal tumors (EGISTs) are rare neoplasms. This study was performed to assess MMP-2 and MMP-9 expression in EGIST tissue samples for association with clinicopathological data from the patients. Twenty-one surgical EGIST tissue specimens were collected for analysis of MMP-2 and MMP-9 expression using immunohistochemistry. MMP-2 and MMP-9 proteins were expressed in all of the epithelial cell types of EGISTs, whereas they were only expressed in 75% of the spindle cell type, although there was no statistically significant difference (p>0.05). Expression of MMP-2 and MMP-9 proteins was associated with tumor size, mitotic rate, tumor necrosis, and distant metastasis (p<0.05). MMP-2 expression was linked with MMP-9 levels (p<0.05). However, there was no correlation between MMP-9 expression and age, sex, primary site, or cell morphology in any of these 21 EGIST patients (p>0.05). Moreover, expression of MMP-2 and MMP-9 proteins increased with the degree of EGIST risk. This study provided evidence of an association of MMP-2 and MMP-9 expression with advanced EGIST behavior.