• Title/Summary/Keyword: SiRNA

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Optimization and Characterization of Conditions for Cellulose-degrading Crude Enzymes Produced by Cellulophaga lytica PKA 1005 (Cellulophaga lytica PKA 1005의 Cellulose 분해 조효소 생산 최적 조건과 조효소의 특성)

  • Bark, Si-Woo;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Pak, Won-Min;Kim, Bo-Ram;Park, Hong-Min;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.18-24
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    • 2014
  • This study was conducted to investigate optimum conditions for the production of cellulose-degrading crude enzymes by an isolated marine bacterium. A marine microorganism producing an extracellular cellulose-degrading enzyme was isolated from the red seaweed, Grateloupia elliptica Holmes. The isolated bacterium was identified as Cellulophaga lytica by 16S ribosomal RNA gene sequence analysis and physiological profiling and designated as Cellulophaga lytica PKA 1005. The optimum conditions for the growth of Cellulophaga lytica PKA 1005 were pH 7, 2% NaCl, and $30^{\circ}C$ with 36 h incubation time. To obtain the crude enzyme, the culture medium of the strain was centrifuged for 30 min at $12,000{\times}g$ and $4^{\circ}C$, and the supernatant was used as crude enzyme. The optimum conditions for the production of the cellulose-degrading crude enzyme were pH 8, $35^{\circ}C$, 8% carboxyl methyl cellulose, and 60 h reaction time.

A Review of Experimental study on Dementia in Oriental medicine;within Oriental medicine journal since 2000 (치매에 대한 최신 실험적 연구 동향;2000년 이후 한의학 학술지를 중심으로)

  • Choi, Sung-Youl;Kim, Dae-Hyun;Kim, Sang-Tae;Kim, Tae-Heon;Kang, Hyung-Won;Lyu, Yeong-Su
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.1
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    • pp.125-146
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    • 2008
  • Objectives : The purpose of this study is to suggest for the following experimental study of dementia by reviewing recent oriental medicine journals that have been published since 2000. Methods: We have investigated various types of studies in relation to dementia through 90 articles that have been published from 2000 to 2007 in recent oriental medicine journals were registered Korea research foundation. Results and Conclusions : 1. Since 2000, 88 articles in relation to dementia have been published and almost of them are herbal medicine-centered studies. Also they show a tendency to increase every year. The journal of oriental neuropsychiatry carries the highest number of studies in relation to dementia. 2. According to the experimental paper, there are 30 cases of using herb simplexes, 48 cases of herb-combined prescription, and 10 cases of other ways. Especially 7 cases of using herb-combined prescription relation to Sasang constitution are all for the Taeumin. 3. There are 85 cases of Animal and cellular experimental, 60 cases of using pathologic model induced cytotoxic activity, a case of using L-NAME, 3 cases of 192 saporin, 4 cases of ibotenic acid, 10 cases of focal cerebral ischemia, 3 cases of alcohol-administered, and one case of natural degradation. 4. Moms water maze, Radial arm maze Passive avoidance learning model were using for examining learning and memory of model animal 5. We propose that following studies of dementia are to he investigated of the applied method of using siRNA with tranceduced gene, sample preparation by water-soaking, oriental medical diagnosis, standardization of differentiating symptom and herb simplexes, building the database by classified prescriptions, and experiment model which are based on precise examining mechanism with cell line as like mouse H19-7 hippocampus, rat HT22 hippocampus, astrocyte, microglia, using the model of animals at APP, PS1, BACE, CT99/PS1, APOE4, Tau, APP/PSI/Tau

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Cinnamaldehyde Derivatives Inhibit Coxsackievirus B3-Induced Viral Myocarditis

  • Li, Xiao-Qiang;Liu, Xiao-Xiao;Wang, Xue-Ying;Xie, Yan-Hua;Yang, Qian;Liu, Xin-Xin;Ding, Yuan-Yuan;Cao, Wei;Wang, Si-Wang
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.279-287
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    • 2017
  • The chemical property of cinnamaldehyde is unstable in vivo, although early experiments have shown its obvious therapeutic effects on viral myocarditis (VMC). To overcome this problem, we used cinnamaldehyde as a leading compound to synthesize derivatives. Five derivatives of cinnamaldehyde were synthesized: 4-methylcinnamaldehyde (1), 4-chlorocinnamaldehyde (2), 4-methoxycinnamaldehyde (3), ${\alpha}$-bromo-4-methylcinnamaldehyde (4), and ${\alpha}$-bromo-4-chlorocinnamaldehyde (5). Neonatal rat cardiomyocytes and HeLa cells infected by coxsackievirus B3 (CVB3) were used to evaluate their antiviral and cytotoxic effects. In vivo BALB/c mice were infected with CVB3 for establishing VMC models. Among the derivatives, compound 4 and 5 inhibited the CVB3 in HeLa cells with the half-maximal inhibitory concentrations values of $11.38{\pm}2.22{\mu}M$ and $2.12{\pm}0.37{\mu}M$, respectively. The 50% toxic concentrations of compound 4 and 5-treated cells were 39-fold and 87-fold higher than in the cinnamaldehyde group. Compound 4 and 5 effectively reduced the viral titers and cardiac pathological changes in a dose-dependent manner. In addition, compound 4 and 5 significantly inhibited the secretion, mRNA and protein expressions of inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in CVB3-infected cardiomyocytes, indicating that brominated cinnamaldehyde not only improved the anti-vital activities for VMC, but also had potent anti-inflammatory effects in cardiomyocytes induced by CVB3.

Adenosine A3 Receptor Mediates ERK1/2- and JNK-Dependent TNF-α Production in Toxoplasma gondii-Infected HTR8/SVneo Human Extravillous Trophoblast Cells

  • Ye, Wei;Sun, Jinhui;Li, Chunchao;Fan, Xuanyan;Gong, Fan;Huang, Xinqia;Deng, Mingzhu;Chu, Jia-Qi
    • Parasites, Hosts and Diseases
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    • v.58 no.4
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    • pp.393-402
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    • 2020
  • Toxoplasma gondii is an intracellular parasite that causes severe disease when the infection occurs during pregnancy. Adenosine is a purine nucleoside involved in numerous physiological processes; however, the role of adenosine receptors in T. gondii-induced trophoblast cell function has not been investigated until now. The goal of the present study was to evaluate the intracellular signaling pathways regulated by adenosine receptors using a HTR-8/SVneo trophoblast cell model of T. gondii infection. HTR8/SVneo human extravillous trophoblast cells were infected with or without T. gondii and then evaluated for cell morphology, intracellular proliferation of the parasite, adenosine receptor expression, TNF-α production and mitogen-activated protein (MAP) kinase signaling pathways triggered by adenosine A3 receptor (A3AR). HTR8/SVneo cells infected with T. gondii exhibited an altered cytoskeletal changes, an increased infection rate and reduced viability in an infection time-dependent manner. T. gondii significantly promoted increased TNF-α production, A3AR protein levels and p38, ERK1/2 and JNK phosphorylation compared to those observed in uninfected control cells. Moreover, the inhibition of A3AR by A3AR siRNA transfection apparently suppressed the T. gondii infection-mediated upregulation of TNF-α, A3AR production and MAPK activation. In addition, T. gondii-promoted TNF-α secretion was dramatically attenuated by pretreatment with PD098059 or SP600125. These results indicate that A3AR-mediated activation of ERK1/2 and JNK positively regulates TNF-α secretion in T. gondii-infected HTR8/SVneo cells.

Hypoxia Mediates Runt-Related Transcription Factor 2 Expression via Induction of Vascular Endothelial Growth Factor in Periodontal Ligament Stem Cells

  • Xu, Qian;Liu, Zhihua;Guo, Ling;Liu, Rui;Li, Rulei;Chu, Xiang;Yang, Jiajia;Luo, Jia;Chen, Faming;Deng, Manjing
    • Molecules and Cells
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    • v.42 no.11
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    • pp.763-772
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    • 2019
  • Periodontitis is characterized by the loss of periodontal tissues, especially alveolar bone. Common therapies cannot satisfactorily recover lost alveolar bone. Periodontal ligament stem cells (PDLSCs) possess the capacity of self-renewal and multilineage differentiation and are likely to recover lost alveolar bone. In addition, periodontitis is accompanied by hypoxia, and hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) is a master transcription factor in the response to hypoxia. Thus, we aimed to ascertain how hypoxia affects runt-related transcription factor 2 (RUNX2), a key osteogenic marker, in the osteogenesis of PDLSCs. In this study, we found that hypoxia enhanced the protein expression of $HIF-1{\alpha}$, vascular endothelial growth factor (VEGF), and RUNX2 ex vivo and in situ. VEGF is a target gene of $HIF-1{\alpha}$, and the increased expression of VEGF and RUNX2 proteins was enhanced by cobalt chloride ($CoCl_2$, $100{\mu}mol/L$), an agonist of $HIF-1{\alpha}$, and suppressed by 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1, $10{\mu}mol/L$), an antagonist of $HIF-1{\alpha}$. In addition, VEGF could regulate the expression of RUNX2, as RUNX2 expression was enhanced by human VEGF ($hVEGF_{165}$) and suppressed by VEGF siRNA. In addition, knocking down VEGF could decrease the expression of osteogenesis-related genes, i.e., RUNX2, alkaline phosphatase (ALP), and type I collagen (COL1), and hypoxia could enhance the expression of ALP, COL1, and osteocalcin (OCN) in the early stage of osteogenesis of PDLSCs. Taken together, our results showed that hypoxia could mediate the expression of RUNX2 in PDLSCs via $HIF-1{\alpha}$-induced VEGF and play a positive role in the early stage of osteogenesis of PDLSCs.

Triptolide Inhibits Lipopolysaccharide-Induced MUC5AC/5B Expression via Nuclear Factor-Kappa B in Human Airway Epithelial Cells (사람 호흡기 상피세포에서 Triptolide의 Nuclear Factor-Kappa B를 통한 Lipopolysaccharide로 유도된 MUC5AC/5B 발현 억제 효과)

  • Seo, Bo Hyeon;Choi, Tae Yeong;Choi, Yoon Seok;Bae, Chang Hoon;Na, Hyung Gyun;Song, Si-Youn;Kim, Yong-Dae
    • Korean Journal of Otorhinolaryngology-Head and Neck Surgery
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    • v.61 no.12
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    • pp.674-680
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    • 2018
  • Background and Objectives The representative mucin genes in the human airway are MUC5AC and MUC5B, which are regulated by several inflammatory and anti-inflammatory substances. Triptolide (TPL), udenafil, betulinic acid, changkil saponin, and glucosteroid are some of the many anti-inflammatory substances that exist. TPL is a diterpenoid compound from the thunder god vine, which is used in traditional Chinese medicine for treatment of immune inflammatory diseases, such as rheumatoid arthritis, systemic lupus erythematosus, nephritis and asthma. However, the effects of TPL on mucin expression of human airway epithelial cells have yet to be reported. Hence, this study investigated the effect of TPL on lipopolysaccharide (LPS)-induced MUC5AC and MUC5B expression in human airway epithelial cells. Subjects and Method The NCI-H292 cells and the primary cultures of human nasal epithelial cells were used to investigate the effects of TPL on LPS-induced MUC5AC and MUC5B expression using real-time polymerase chain reaction, enzyme immunoassay, and Western blot. Results TPL significantly decreased the LPS-induced MUC5AC and MUC5B mRNA expression and protein production. TPL also significantly decreased the nuclear factor-kappa B (NF-kB) phosphorylation. Conclusion These results suggest that TPL down regulates MUC5AC and MUC5B expression via inhibition of NF-kB activation in human airway epithelial cells. This study may provide important information about the biological role of triptolide on mucus-secretion in airway inflammatory diseases and the development of novel therapeutic agents for controlling such diseases.

The oral microbiome of implant-abutment screw holes compared with the peri-implant sulcus and natural supragingival plaque in healthy individuals

  • MinKee Son;Yuri Song;Yeuni Yu;Si Yeong Kim;Jung-Bo Huh;Eun-Bin Bae;Won-Tak Cho;Hee Sam Na;Jin Chung
    • Journal of Periodontal and Implant Science
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    • v.53 no.3
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    • pp.233-244
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    • 2023
  • Purpose: An implant-supported prosthesis consists of an implant fixture, an abutment, an internal screw that connects the abutment to the implant fixture, and the upper prosthesis. Numerous studies have investigated the microorganisms present on the implant surface, surrounding tissues, and the subgingival microflora associated with peri-implantitis. However, there is limited information regarding the microbiome within the internal screw space. In this study, microbial samples were collected from the supragingival surfaces of natural teeth, the peri-implant sulcus, and the implant-abutment screw hole, in order to characterize the microbiome of the internal screw space in healthy subjects. Methods: Samples were obtained from the supragingival region of natural teeth, the peri-implant sulcus, and the implant screw hole in 20 healthy subjects. DNA was extracted, and the V3-V4 region of the 16S ribosomal RNA was sequenced for microbiome analysis. Alpha diversity, beta diversity, linear discriminant analysis effect size (LEfSe), and network analysis were employed to compare the characteristics of the microbiomes. Results: We observed significant differences in beta diversity among the samples. Upon analyzing the significant taxa using LEfSe, the microbial composition of the implant-abutment screw hole's microbiome was found to be similar to that of the other sampling sites' microbiomes. Moreover, the microbiome network analysis revealed a unique network complexity in samples obtained from the implant screw hole compared to those from the other sampling sites. Conclusions: The bacterial composition of the biofilm collected from the implant-abutment screw hole exhibited significant differences compared to the supra-structure of the implant. Therefore, long-term monitoring and management of not only the peri-implant tissue but also the implant screw are necessary.

Cordycepin Inhibits LPS-induced Cell Migration and Invasion in Human Colorectal Carcinoma HCT116 cells through Down-regulation of Prostaglandin E2-EP4 Receptor (LPS 유도된 HCT116 인간 대장암세포에서 cordycepin의 prostaglandin E2-EP4 receptor 감소 조절을 통한 세포의 이동과 전이 억제 효과)

  • Jung Eun Kim;Bo-Ram Kim;Su Hui Seong;Jin-Ho Kim;Ha-Nul Lee;Chan Seo;Ji Min Jung;Su A Im;Kyung-Min Choi;Jin-Woo Jeong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2023.04a
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    • pp.50-50
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    • 2023
  • Prostaglandin E2(PGE2), a major product of cyclooxygenase-2 (COX-2), plays an important role in the carcinogenesis of many solid tumors, including colorectal cancer. Because PGE2 functions by signaling through PGE2 receptors (Eps), which regulate tumor cell growth, invasion, and migration, there has been a growing amount of interest in the therapeutic potential of targeting Eps. In the present study, we investigated the role of EP4 on the effectiveness of cordycepin in inhibititing the migration and invasion of HCT116 human colorectal carcinoma cells. Our data indicate that cordycepin suppressed lipopolysaccharide (LPS)-enhanced cell migration and invasion through the inactivation of matrix metalloproteinases (MMP)-9 as well as the down-regulation of COX-2 expression and PGE2 production. These events were shown to be associated with the inactivation of EP4 and activation of AMP-activated protein kinase (AMPK). Moreover, the AMPK inhibitor, compound C, as well as AMPK knockdown via siRNA, attenuated the cordycepin-induced inhibition of EP4 expression. Cordycepin treatment also reduced the activation of CREB. These findings indicate that cordycepin suppresses the migration and invasion of HCT116 cells. Through modulating EP4 expression and the AMPK-CREB signaling pathway. Therefore, cordycepin has the potential to serve as a potent anti-cancer agent in therapeutic strategies against colorectal cancer metastasis.

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Valproic Acid-induced PPAR-alpha and FGF21 Expression Involves Survival Response in Hepatocytes (Valproic acid에 의해 증가하는 PPAR-alpha 및 FGF21의 발현이 간세포 생존에 미치는 영향)

  • Bakhovuddin Azamov;Yeowon Kang;Chanhee Lee;Wan-Seog Shim;Kwang Min Lee;Parkyong Song
    • Journal of Life Science
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    • v.34 no.4
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    • pp.227-235
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    • 2024
  • Hepatocyte damage caused by medications or herbal products is one of the important problem when these compounds are chronically administrated. Thus, improving hepatocyte survival during treatment offers a wide range of opportunities. Valproic acid (VPA), a branched short-chain fatty acid derived from naturally occurring valeric acid, is commonly used to treat epilepsy and seizures. Although VPA exerts numerous effects in cancer, HIV therapy, and neurodegenerative disease, its effects on the liver and its mechanism of action have not been fully elucidated. Here, we demonstrated that VPA caused moderate liver cell toxicity and apoptosis. Interestingly, VPA treatment increased transcription levels of PPAR alpha (PPAR-α) and fibroblast growth factor 21 (FGF21) in murine (Hepa1c1c7) hepatoma cells in a time and concentration dependent manner. VPA-induced FGF21 expression was significantly weaker under PPAR-α silencing condition than in cells transfected with non-targeting control siRNA. Subsequent experiments showed that cell viability was significantly lowered when the FGF21 signaling pathway was blocked by FGF receptor antagonist. Finally, we further determined that AMPK phosphorylation was not responsible for VPA-induced FGF21 expression and PPAR-a increments. These results indicate that increases of FGF21 expression alleviate VPA-induced hepatic toxicity, thereby making FGF21 a potential biomarker for predicting liver damage during VPA treatments.

Effects of the Cell-to-Cell Communication between Oocyte and Cumulus Cells on the Quality of Oocytes (난자와 난구세포간 Cell-to-Cell Communication이 난자에 미치는 영향)

  • 신창숙;윤세진;박창은;이경아
    • Development and Reproduction
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    • v.5 no.2
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    • pp.115-122
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    • 2001
  • Production of a mature oocyte is a complex process that requires the close association between oocyte and follicular cells. The present study was conducted to investigate the difference between oocytes with and without close junctional communication with cumulus cells and the involvement of two connexins(CXs) in the interactions. Follicles at different sizes(small: 200~400 ㎛; large:>450 ㎛) were mechanically isolated from PMSG-primed mouse ovaries, and punctured to get cumulus-oocyte complex(COC). Oocytes were released themselves(denuded), with partially attached(partial), or with tightly attached(intact) cumulus cells. Maturation and fertilization capacity of the COCs were measured. Expression of CX 37 and CX 43 was examined by RT-PCR and in situ hybridization. The ratio between intact COC and denude/partial oocytes was 30%(SI) and 70%(SPD) in small follicles, while 55%(LI) and 45%(LPD) in large follicles, respectively. Maturation and fertilization rates of the released oocytes were similar among SI, LPD, LI groups, but those were significantly lower in SPD oocytes. In oocytes, CX 37 was the major CX and CX 43 was not expressed, whereas in the cumulus cells, CX 43 was the major, and CX 37 was the next. Results of the present study suggest that 1) immature oocytes from small follicles with intact cell-to-cell communication with cumulus have the similar quality to that of the oocytes from larger follicles, 2) gap junction between oocyte and cumulus cells may be the heterotypic channel, and 3) we could not explain the difference in the cell-to-cell communication between intact and partial/denuded COCs through the expression of the two CXs.

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