• Korean Journal of Veterinary Research
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• v.44 no.2
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• pp.163-169
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• 2004

To elucidate the molecular mechanisms of autoimmune inflammation in the central nervous system, we examined the expression and localization of STAT1, STAT3, STAT4 and STAT6 molecules during experimental autoimmune encephalomyelitis (EAE) by competitive PCR. In the present study, we quantitated IL-4 and IL-12 p40 mRNA by competitive PCR in the CNS during EAE. IL-4 mRNA was found at early and peak stages. On the other hand, the IL-12 p40 mRNA level reached maximal levels at the peak stage and still found at the recovery stage of the disease. We examined the kinetics of STAT mRNA in the CNS during EAE and demonstrated that STAT1 and STAT4 mRNA reached a maximal level at the peak stage of EAE, whereas STAT3 mRNA level increased gradually to the recovery stage. STAT6 mRNA increased rapidly at the early stage followed by gradual decrease till the recovery stage. Taken together, these findings suggest that STAT4 which was probably activated by IL-12 plays a pro-inflammatory role and that STAT3 which was activated throughout the disease course seems to serve as a transducer of anti-inflammatory signals.

• Journal of Animal Science and Technology
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• v.53 no.5
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• pp.419-428
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• 2011

This study was conducted to test the efficacy of Ulva pertusa kjellman as immunomodulators under lipopolysaccharide (LPS) challenge in broilers by investigating their effects on serum superoxide dismutase (SOD) like ability, immunoglobulin concentration, and splenic cytokine mRNA expression. A total of ninety six1-d-old male broiler chicks (Ross 308) were divided into 4 treatment groups with 8 replicates of 3 birds in each group. NC (negative control, no immune substances), PC (positive control, ${\beta}$-glucan 25 ppm), Ulva P (Ulva pertusa kjellman Powder 3%), and Ulva E (Extract form Ulva pertusa kjellman 0.3%) were added in feed with respective substance. Heparinized venous blood and spleens were collected from five birds per dietary treatment at 5 wk of age. The SOD like activities in Ulva P and Ulva E were significantly increased in comparison with other groups (P<0.05). The immunoglobulin M content was lower in the Ulva E than other groups (P<0.05). Expression patterns of splenic cytokine mRNA in the IL-$1{\beeta}$, IL-2 and IL-6 were similar. Expression rate of IL-$1{\beta}$, IL-2 and IL-6 in Ulva pertusa kjellman (Ulva P, Ulva E) were decreased (P<0.05) in comparison to other groups. Expression rate of IL-$1{\beta}$, IL-2 and IL-6 were significantly lower in groups treated with Ulva E than Ulva P. In conclusion, dietary supplementation of Ulva pertusa kjellman improved SOD like activity and affect immune system by inhibiting inflammatory response in broiler chicks. In addition, it is more effective to use Ulva pertusa kjellman extract than powder for immunomodulatory function. These results suggest the possibility that Ulva pertusa kjellman could be used as the immunomodulator for inflammatory response in broiler chicks.

• Korean Journal of Pharmacognosy
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• v.30 no.4
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• pp.355-362
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• 1999

New lectins have been isolated and purified from mung bean (Phaseolus radiatus) through physiological saline extraction, ammonium sulfate salt fractionation and column chromatographies. Ion exchanger were eluted by linear salt gradient and then further purified through gel filtration. Thus obtained lectin named as MBL. The gene expressions of 5 cytokines (IL-1, IL-2, IL-6, $TNF-{\aphpa}$ and $IFN-{\gamma}$) from human peripheral blood mononuclear cells (PBMC) stimulated with MBL were investigated by using reverse transcription polymerase chain reaction (RT-PCR). PBMC ($1{\times}106$ cells/ml) isolated from healthy volunteers were stimulated with lectins (4 mg/ml) for various time intervals (1 to 96 hrs). After each of the various stimulated times, total RNA was isolated and assessed for different cytokines mRNA by RT-PCR. The mRNA encoding IL-1, IL-2 were detected continuously from 1 to 20 hrs, and IL-6 was detected up to 24 hrs. But the mRNA encoding $IFN-{\gamma}$ and $TNF-{\alpha}$ were detected to 8 hours only and showed short time response compared with other cytokines. The significant expression of all cytokines mRNA were observed at 4 hrs. These results suggested that MBL, as inducer of cytokines could elicit detectable cytokine mRNA from PBMC.

• Journal of Veterinary Clinics
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• v.27 no.5
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• pp.508-513
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• 2010

Cytokines are important mediators of the immune response, and quantitating cytokine mRNA is a highly sensitive and attractive method for measuring cytokine production. The objective of the current study was to develop and validate a SYBR green quantitative real-time reverse transcriptase PCR (qRT-PCR) assay for measuring canine cytokine mRNA. The optimal annealing temperatures ($T_a$) of the designed primers were $62^{\circ}C$ for interleukin (IL)-$1{\beta}$, IL-6 and IL-10; $60^{\circ}C$ for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and tumor necrosis factor (TNF)-${\alpha}$; and $58^{\circ}C$ for high mobility group box 1 (HMGB1). Primer efficiencies of all primers calculated for standard curve samples were between 97.1% and 102.6%. No evidence of secondary structure or primer-dimer formation was seen via melt-curve analysis or gel electrophoresis. The developed qRT-PCR assays are highly specific and sensitive and can be used to quantify gene expression levels of canine cytokines.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.10
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• pp.1329-1335
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• 2006

LP-BM5 murine leukemia retrovirus induces the excessive oxidative stress and immune dysfunction leading to B cell leukemia and murine AIDS with cytokine dysfunction. In the present study, the immune restoratory effect of antioxidant hormone dedydroepiandrosterone sulfate (DHEAS) was investigated in the primary splenocytes from LP-BM5 retrovirus-infected C57BL/6 mice. DHEAS significantly increased T and B cell response to mitogen and normalized the unbalanced production of Th1/Th2 type cytokines. In particular, both protein and mRNA expression of IL-4, IL-6, and $TNF-\alpha$ were down-regulated by DHEAS treatment whereas IL-2 and $IFN-\gamma$ level were increased. This result suggests that DHEAS directly or indirectly regulates the gene expression of Th1/Th2 type cytokines in transcription level. In addition, DHEAS treatment decreased the hepatic lipid peroxidation and preserved vitamin E level in liver cells. These results suggested that DHEAS could effectively prevent immune dysfunction by regulating cytokine secretion and preventing the oxidative stress in murine AIDS.

• CELLMED
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• v.7 no.3
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• pp.15.1-15.6
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• 2017

We analyzed the effects of an Atopic Preventive Drink (APD) on the regulation of Th2 cytokines using RAW 264.7 macrophage cells. In the evaluation of nitric oxide (NO) production in cells, NO production levels were shown to be elevated only in the APD-treated group in a dose-dependent manner. In the lipopolysaccharide (LPS) with APD-treated group, NO production significantly decreased as APD concentration increased. Further, mRNA expression levels and protein concentrations of pro-inflammatory cytokines in cells were determined. Th2 stimulatory cytokine ($IL-1{\beta}$) and Th2 cytokine (IL-6 and IL-10) levels were significantly reduced in the LPS with APD-treated group compared to the only LPS-treated group. mRNA expression levels of inflammatory-related genes (COX-2 and iNOS) were significantly reduced in the LPS with APD-treated group compared to the only LPS-treated group. These results suggest that APD has an anti-atopic effect by reducing mRNA and proteins expressions of Th2 cytokines and inflammatory-related genes.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.77-84
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• 2003

Chitinase was purified from an antagonistic bacterium Bacillus sp. 7079 by ammonium sulfate precipitation, QAE-Sephadex anion exchange chromatography, Sephadex G-100 gel filtration, and SP-Sephadex cation exchange chromatography. The molecula. weight of purified chitinase (PC-1) was approximately 66.5 kDa on SDS-PACE. PC-1 exhibited optimum pH and temperature of pH 7.5 and $45^{\circ}C$, respectively. More than $80\%$ of PC-1 was stable at pH 5.0 to 9.0, and more than $90\%$ at $40^{\circ}C$. $Fe^2+\;and\;Ca^2+$ inhibited the chitinase activity about $20\%$, and EDTA and p-CMB by about $30\%$, whereas $Ag^+$ inhibited the activity up to $65\%$. The $K_m$ value of PC-1 was 1.215 mg/ml with colloidal chitin as a substrate. We also investigated the effect of PC-1 treated chitin (PCTC) on the pro-inflammatory cytokine gene expression in macrophage RAW 264.7 cells. The expression of IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA gene was investigated using reverse transcriptase polymerase chain reaction (RT-PCR). IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were induced by the treatment of PCTC and chitin only in RAW 264.7 cells. These expressions were induced as early as 2 h and sustained up to 24 h in RAW 264.7 cells. IL-$1{\alpha}$ and IL-$1{\beta}$ mRNA were more strongly expressed by the treatment of PCTC than chitin treatment alone in RAW 264.7 cells.

• YAKHAK HOEJI
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• v.46 no.6
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• pp.472-476
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• 2002

Contact hypersensitivity (CHS) serves as a good model of cell-mediated reaction. Epidermal langerhans cell (LC) are thought to playa crucial role in the regulation of immune reaction of the skin, which elicit the CHS response by presenting Antigen to trafficking Ag-specific T cells within the skin. However, contact hypersensitivity is regarded as a negative side of immunities, caused by increased damaging immune response. Therefore, the study of effector molecule causing immune suppression is thought to be meaningful in the skin immune response. For this aim, this study investigated the influence of pedunculagin on cytokine, IL-$\beta$ expression from langerhans cell (LC). In vitro and in vivo, pedunculagin up-regulated the expression of IL-1$\beta$ mRNA. After PMA stimulation in vitro and DNFB sensitization in vivo, the expression of IL-1$\beta$ mRNA was down-regulated. This results suggested that pedunculagin could be immuno-modulator in skin immune system by modulating IL-1$\beta$ expression.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.5
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• pp.413-420
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• 2015

Dexmedetomidine is a sedative and analgesic agent that exerts its effects by selectively agonizing ${\alpha}2$ adrenoceptor. Histamine is a pathophysiological amine that activates G protein-coupled receptors, to induce $Ca^{2+}$ release and subsequent mediate or progress inflammation. Dexmedetomidine has been reported to exert inhibitory effect on inflammation both in vitro and in vivo studies. However, it is unclear that dexmedetomidine modulates histamine-induced signaling and pro-inflammatory cytokine expression. This study was carried out to assess how dexmedetomidine modulates histamine-induced $Ca^{2+}$ signaling and regulates the expression of pro-inflammatory cytokine genes encoding interleukin (IL)-6 and -8. To elucidate the regulatory role of dexmedetomidine on histamine signaling, HeLa cells and human salivary gland cells which are endogenously expressed histamine 1 receptor were used. Dexmedetomidine itself did not trigger $Ca^{2+}$ peak or increase in the presence or absence of external $Ca^{2+}$. When cells were stimulated with histamine after pretreatment with various concentrations of dexmedetomidine, we observed inhibited histamine-induced $[Ca^{2+}]_i$ signal in both cell types. Histamine stimulated IL-6 mRNA expression not IL-8 mRNA within 2 hrs, however this effect was attenuated by dexmedetomidine. Collectively, these findings suggest that dexmedetomidine modulates histamine-induced $Ca^{2+}$ signaling and IL-6 expression and will be useful for understanding the antagonistic properties of dexmedetomidine on histamine-induced signaling beyond its sedative effect.

• The Journal of Internal Korean Medicine
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• v.24 no.2
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• pp.249-259
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• 2003

Objective : We aimed to identify the inhibitory effects of SocheongRyoungTang on Cytokine Production and Secretion of IgE in Highly Purified Mouse B cells. This experiment was designed to investigate the effect of SoCheongRyongTang(SCRT) on Antiallergy. Materials and Methods : We measured the cytotoxic activity for cytokines transcript expression, production of $IL-1{\beta},\;IL-4,\;IL-6,\;IL-10,\;IFN-{\gamma},\;TNF-{\alpha},\;TGF-{\beta}$ proliferation of B cell in anti-CD40mAb plus rIL-4 plus HRF stimulated murine splenic B cells and histamine in anti-CD40mAb plus rIL-4 plus HRF stimulated mast cells. Results : 1. SCRT increased the gene synthesis of $IFN-{\gamma}(m-RNA)$, the appearance of IL-10, $IFN-{\gamma}$. 2. SCRT decreased the gene synthesis of $IL-l{\beta},\;IL-4,\;TGF-{\beta}(m-RNA)$ and the appearance of $IL-l{\beta},\;IL-4,\;TGF-{\beta},\;IgE$ significantly. Conclusion : SCRT decreased the proliferation of B cell significantly. According to the above results, it is suggested that SCRT extract might be usefully applied for prevention and treatment of Allergic disease.