• Title/Summary/Keyword: E6/E7 mRNA

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Effect of Sanyeoleumja on Inflammatory Response of RAW 264.7 Cells (RAW 264.7 cell의 염증반응에 대한 산열음자(散熱飮子)의 항염증 효과)

  • Kim, Tae Yeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.1
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    • pp.7-13
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    • 2020
  • Sanyeoleumja (SY) is the traditional Korean medicinal prescription for the treatment of inflammatory diseases of eyes. In this study, the anti-inflammatory effects of SY water extract were investigated. To measure the anti-inflammatory effects of SY, we examined the productions of inflammatory factor including nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), interleukin-1β (IL-1β) in lipopolysaccharide (LPS)-induced RAW 264.7 cells. SY inhibited NO and PGE2 production in a dose dependent manner and decreased the protein and mRNA expression of iNOS and COX-2. Also, SY decreased the mRNA expression of interleukin-6 (IL-6) and interleukin-1β (IL-1β). In conclusion, SY downregulated LPS-induced inflammatory factor productions, which could be a clinical basis for inflammatory diseases.

Anti-inflammatory effects of ethanolic mulberry extract on the murine macrophage cell line, RAW 264.7 (RAW 264.7 큰포식세포에서 상백피 및 상지 에탄올 추출물의 항염증 활성 비교)

  • Kim, Yunyoung;Yang, Yoon Kyoung;Kim, Dongmin;Kim, Ji Yeon
    • Korean Journal of Food Science and Technology
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    • v.49 no.3
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    • pp.343-348
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    • 2017
  • The aim of this study was to compare the anti-inflammatory effects of ethanol extracts of root peel and spear of mulberry (RME and SME, respectively) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Both extracts significantly inhibited the production of reactive oxygen species (ROS), nitric oxide (NO), and interleukin-6 (IL-6). However, prostaglandin $E_2$ ($PGE_2$) levels decreased in LPS-stimulated RAW 264.7 cells treated with SME. Additionally, the extracts reduced inducible NO synthase (iNOS) expression and cyclooxygenase-2 (COX-2) in mRNA levels. Although ROS production was lower in the RME-treated cells than in the SME-treated cells, the levels of other inflammatory parameters, including IL-6 and $PGE_2$, and mRNA levels of iNOS and COX-2 reduced more in the SME-treated cells. These results indicate that SME showed higher anti-inflammatory activities than RME. Therefore, SME can be used as a functional food ingredient to enhance health.

Microbial Community Diversity in Anaerobic Reactors Digesting Turkey, Chicken, and Swine Wastes

  • Ziganshina, Elvira E.;Belostotskiy, Dmitry E.;Shushlyaev, Roman V.;Miluykov, Vasili A.;Vankov, Petr Y.;Ziganshin, Ayrat M.
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1464-1472
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    • 2014
  • The microbial community structures of two continuous stirred tank reactors digesting turkey manure with pine wood shavings as well as chicken and swine manure were investigated. The reactor fed with chicken/swine wastes displayed the highest organic acids concentration (up to 15.2 g/l) and ammonia concentration (up to 3.7 g/l ammonium nitrogen) and generated a higher biogas yield (up to $366ml/g_{VS}$) compared with the reactor supplied with turkey wastes (1.5-1.8 g/l of organic acids and 1.6-1.7 g/l of ammonium levels; biogas yield was up to $195ml/g_{VS}$). The microbial community diversity was assessed using both sequencing and profiling terminal restriction fragment length polymorphisms of 16S rRNA genes. Additionally, methanogens were analyzed using methyl coenzyme M reductase alpha subunit (mcrA) genes. The bacterial community was dominated by members of unclassified Clostridiales with the prevalence of specific clostridial phylotypes in each reactor, indicating the effect of the substrate type on the community structure. Of the methanogenic archaea, methanogens of the genus Methanosarcina were found in high proportions in both reactors with specific methanosarcinas in each reactor, whereas the strict hydrogenotrophic methanogens of Methanoculleus sp. were found at significant levels only in the reactor fed with chicken/swine manure (based on the analyses of 16S rRNA gene). This suggests that among methanogenic archaea, Methanosarcina species which have different metabolic capabilities, including aceticlastic and hydrogenotrophic methanogenesis, were mainly involved in anaerobic digestion of turkey wastes.

Expression of Hepatic Vascular Stress Genes Following Ischemiai/Reperfusion and Subsequent Endotoxemia

  • Kim, Sung-Ho;Lee, Sun-Mee
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.769-775
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    • 2004
  • Hepatic ischemia and reperfusion (l/R) predisposes the liver to secondary stresses such as endotoxemia, possibly via dysregulation of the hepatic microcirculation secondary to an imbalanced regulation of the vascular stress genes. In this study, the effect of hepatic I/R on the hepatic vasoregulatory gene expression in response to endotoxin was determined. Rats were subjected to 90 min of hepatic ischemia and 6 h of reperfusion. Lipopolysaccharide (LPS, 1 mg/kg) was injected intraperitoneally after reperfusion. Plasma and liver samples were obtained 6 h after reperfusion for serum aminotransferase assays and RT-PCR analysis of the mRNA for the genes of interest: endothelin-1 (ET-1), its receptors $ET_A$ and $ET_B$, endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), cyciooxygenase-2 (COX-2), and tumor necrosis factor-a (TNF-${\alpha}$). The activities of serum aminotransferases were significantly increased in the I/R group. This increase was markedly potentiated by LPS treatment. The ET-1 mRNA was increased by LPS alone, and this increase was significantly greater in both the I/R alone and I/R + LPS groups compared to the sham. There were no significant differences in ETA receptor mRNA levels among any of the experimental groups. $ET_B$ mRNA was increased by both LPS alone and I/R alone, with no significant difference between the I/R alone and I/R + LPS groups. The eN OS and HO-1 transcripts were increased by I/R alone and further increased by I/R + LPS. The iNOS mRNA levels were increased by I/R alone, but increased significantly more by both LPS alone and I/R + LPS compared to I/R alone. The TNF-${\alpha}$ mRNA levels showed no change with I/R alone, but were increased by both LPS alone and I/R + LPS. The COX-2 expression was increased significantly by I/R alone and significantly more by I/R + LPS. Taken collectively, significantly greater induction of the vasodilator genes over the constriction forces was observed with I/R + LPS. These results may partly explain the increased susceptibility of ischemic livers to injury as a result of endotoxemia.

Inhibitory Effect of Eurya emarginata on the Production of Pro-inflammatory Cytokines in Murine Macrophage RAW264.7 (Murine Macrophage RAW 264.7 세포에서 우묵사스레피에 의한 염증성 사이토카인 억제효과)

  • 박수영;이혜자;현은아;문지영;앙홍철;이남호;김세재;강희경;유은숙
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.311-318
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    • 2003
  • Eurya emarginata (Thunb.) Makino (Theaceae) is distributed in coastal areas of island. The leaves of Eurya are used in the traditional medicine of the coastal areas of jeju island with the aim of diuresis or to treat ulcers. Nevertheless, there are few reports on the biological activity and constituents of E. emarginata. In this study, we investigated the pharmacological activity of the solvent extracts of E. emarginata on the several inflammatory markers (TNF-$\alpha$, IL-1$\beta$, IL-6, NO, iNOS and COX-2). Also we examined the antioxidizing effect of the solvent extracts by determination of DPPH radical-scavenging activity. Among the solvent fractions, EtOAc and BuOH extracts showed potent radical scavenging activity (RC$_{50}$=10.9 and 12.7 respectively). The subtractions of EF 5-4-6-3-2 and BF 1 potentially inhibited the mRNA expression of pro-inflammatory cytokines (IL-1$\beta$, IL-6 and TNF-$\alpha$) at the concentration of 100 $\mu\textrm{g}$/mι. Also the fractions inhibited the mRNA expression of pro-inflammatory cytokines (IL-1$\beta$, IL-6 and TNF-$\alpha$) and protein expression of iNOS and COX-2 at the concentration of 100 $\mu\textrm{g}$/mι. And then, the inhibition of iNOS was correlated with the decrease of nitrite level. These results suggest that E. emarginata may have anti-inflammatory activity through the inhibition of pro-inflammatory cytokines, iNOS and COX-2.2.

Effects of Scutellariae Radix-acupuncture solution On Anti-inflammatory in RAW 264.7 macrophages (황금약침액(黃芩藥針液)이 LPS로 유도(誘導)된 RAW 264.7 대식세포(大食細胞)에서의 항염증효과(抗炎症效果))

  • Choi, Young-Kwang;Park, Sung-Ho;Seo, Il-Bok;Kim, Ho-Hyun;Kim, Jeong-Seon;Kim, Ee-Hwa
    • Journal of Pharmacopuncture
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    • v.6 no.3
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    • pp.5-14
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    • 2003
  • Objective : Recently, Herbal-acupuncture therapeutics has been used for the treatment of inflammatory diseases such as rheumatoid arthritis. Especially, we have been interested in chemical mediators concerned with inflammation such as prostaglandin, cytokine, nitric oxide. The purpose of this study is investigated that the effect of Scutellariae Radix-acupuncture solution in lipopolysaccharide-stimulated RAW 264.7 macrophages, performed several expeimental items : those are prostaglandin E. nitric oxide and cyclooxygenase-2. Method : The cytotoxicity of Scutellariae Radix-acupuncture solution in RAW 264.7 macrophages were measured by MTT-based cytotoxicity assay. In order to observe cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, RT-PCR was used. Prostaglandin $E_2$ formation and nitric oxide production was measured by competitive enzyme immunoassay and Griess assay. Results : 1. The MTT assay demonstrated that cytotoxic effect of Scutellariae Radix-acupuncture solution in RAW 264.7 macrophages were not appeared before concentration of 5mg/mL 2. Scutellariae Radix-acupuncture solution inhibited cyclooxygenase-2 mRNA expression in lipopolysaccharide-treated RAW 264.7 macrophages. 3. Scutellariae Radix-acupuncture solution inhibited nitric oxide production in lipopolysaccharide-treated RAW 264.7 macrophages. 4. Scutellariae Radix-acupuncture solution inhibited prostaglandin $E_2$ formation in lipopolysaccharide-treated RAW 264.7 macrophages. Conclusion : On the basis of these results, It was shown that Scutellariae Radix-acupuncture solution is significantly able to inhibit the production of $PGE_2$ and NO, as well as COX-2 mRNA expression. Our results may provide new mechanism by which Scutellariae Radix-acupuncture solution accounts for its beneficial effect on accelerating wound healing and anti-inflammation.

Biological Activity of Fermented Gryllus bimaculatus extracts (발효 쌍별귀뚜라미 추출물의 생리활성 연구)

  • Park, In-Sun;Lim, Hyeon-Ji;Jeong, Seung-Il;Jung, Chan-Hun
    • Korean Journal of Pharmacognosy
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    • v.52 no.2
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    • pp.92-98
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    • 2021
  • Gryllus bimaculatus extract (GbE) have reported that anti-inflammatory activity by suppression of pro-inflammatory cytokines. However, the effects of fermented Gryllus bimaculatus extract (FGbE) have not yet been investigated. In this study, we evaluated the anti-inflammatory and anti-wrinkle effect of the fermented Gryllus bimaculatus extracts using Bacillus subtilis (JB PMB-18) in RAW264.7 cells. Both GbE and FGbE exerted no cytotoxic effects until 1000 ㎍/mL concentration. FGbE decreased NO production and decreased iNOS and COX-2 mRNA levels in a concentration-dependent manner. In addition, the protein production of inflammatory cytokines TNF-α, IL-1β and IL-6 was effectively reduced compared to the GbE. Inhibitory activities of elastase and collagenase associated with skin wrinkle improvement were measured to be 45% and 69%, respectively, at a concentration of 500 ㎍/mL in FGbE. From these results, FGbE can be used as a health functional food and skin functional cosmetic materials for preventing inflammatory diseases because it has excellent anti-inflammatory and anti-wrinkle effects.

Cloning and Expression of K11 Phage RNA Polymerase (K11 RNA 중합효소의 Cloning 및 발현)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.9 no.1
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    • pp.19-24
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    • 1997
  • Using the PCR(polymerase chain reaction method), gone 1 of phage K11 coding for K11 phage RNA polymerase has been cloned and expressed under the control of lac promoter. K11 phage RNA polymerase was conventionally purified through the DEAE-sephacel and Affigel blue column chromatographies. The 0.2-0.3 M $NH_4Cl$ fractions of DAEA-sephacel column chromatography showed K11 phage RNA polymerase activity and further purification with Affigel blue column chromatography showed nearly single protein band on SDS-polyacryl amide gel. K11 phage RNA polymerase, which is one of the T7 group phage RNA polymerase (E. coil phage T7, T3 and Salmonella tyhimurium phage SP6 RNA polymerase), shares high degrees of homology with the other T7 group phage RNA polymerase. Previously we constructed T7 and SP6 promoter variants and revealed promoter specificity of T7 and SP6 RNA polymerase (Lee and Kang, 1993). To investigate the promoter specificity of K11 RNA polymerase in vitro K11 promoter activity was measured with SP6 promoter variants. The SP6 promoter variant share highest degrees of sequence homology with K11 promoter sequence show strongest promoter activity.

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Effects of Kamiyukgunja-tang on anti-CD40 and Recombinant Interleukin-4 induced Cytokine Production and Immunoglobulin E in Highly Purified Mouse B Cells (생쥐의 B 세포에서 면역글로블린 E의 분비와 사이토카인 생산에 대한 가미육군자탕의 효과)

  • Kim Woon Gil;Kim Dong Hee;Park Yang Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.1065-1074
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    • 2003
  • In order to evaluate the antiallergic effects of Kamiyukgunja-tang (KYGJT), studies were done. We measured the cytotoxic activity for lung fibroblast cell, cytokines transcript expression, production of INF-γ, IL-10, IL-4, GM-CSF, IL-1 β, TNF-α. IL-5 proliferation of B cell in anti-CD40mAb plus r1L-4 stimulated murine splenic B cells. The results were obtained as follows : 1. KYGJT was not showed cytotoxicity in the fibroblast lung cell. 2. KYGJT increased the gene synthesis of INF-γ, IL-10, GM-CSF(m-RNA). 3. KYGJT decreased the gene synthesis of IL-1β, IL-4, TNF-α, IL-5(m-RNA). 4. KYGJT decreased the appearance of TNF-α significantly. 5. KYGJT decreased the appearance of IgE significantly. 6. KYGJT decreased the proliferation of B cell significantly. 7. KYGJT decreased the appearance of Histamin Release Production significantly. The facts above prove that KYGJT is effective against the allergy. Thus. I think that we should study on this continuously

Purification and Characterization of Cop, a Protein Involved in the Copy Number Control of Plasmid pE194

  • Kwak, Jin-Hwan;Kim, Jung-Ho;Kim, Mu-Yong;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.291-297
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    • 1998
  • Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

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