• Korean Journal of Agricultural Science
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• v.41 no.3
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• pp.237-243
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• 2014

Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

• Journal of Veterinary Science
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• v.21 no.1
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• pp.6.1-6.12
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• 2020

Currently, the optimal resuscitation fluid remains debatable. Therefore, in the present study, we designed a trometamol-balanced solution (TBS) for use as a resuscitation fluid for hemorrhagic shock. Hemorrhagic shock was induced in 18 male Wistar-Kyoto rats, which were assigned to normal saline (NS), Ringer's solution (RS), and TBS groups. During the hemorrhagic state, their hemodynamic parameters were recorded using an Abbott i-STAT analyzer with the CG4+ cartridge (for pH, pressure of carbon dioxide, pressure of oxygen, total carbon dioxide, bicarbonate, base excess, oxygen saturation, and lactate), the CG6+ cartridge (for sodium, potassium, chloride, blood glucose, blood urea nitrogen, hematocrit, and hemoglobin), and enzyme-linked immunosorbent assay kits (calcium, magnesium, creatinine, aspartate aminotransferase, alanine aminotransferase, bilirubin, and albumin). Similar trends were found for the parameters of biochemistries, electrolytes, and blood gas, and they revealed no significant changes after blood withdrawal-induced hemorrhagic shock. However, the TBS group showed more effective ability to correct metabolic acidosis than the NS and RS groups. TBS was a feasible and safe resuscitation solution in this study and may be an alternative to NS and RS for resuscitation in hemorrhagic shock patients without liver damage.

• BMB Reports
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• v.37 no.4
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• pp.507-514
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• 2004

Gamma irradiation ($\gamma$-IR) is reported to have diverse effects on immune cell apoptosis, survival and differentiation. In the present study, the immunomodulatory effect of a low dose $\gamma$-IR (5~10 Gy) was investigated, focusing on the role of NF-${\kappa}B$ in the induction of the B cell differentiation molecule, CD23/FceRII. In the human B cell line Ramos, $\gamma$-IR not only induced CD23 expression, but also augmented the IL-4-induced surface CD23 levels. While $\gamma$-IR did not cause STAT6 activation in these cells, it did induce both DNA binding and the transcriptional activity of NF-${\kappa}B$ in the $I{\kappa}B$ degradation-dependent manner. It was subsequently found that different NF-${\kappa}B$ regulating signals modulated the $\gamma$-IR-or IL-4-induced CD23 expression. Inhibitors of NF-${\kappa}B$ activation, such as PDTC and MG132, suppressed the $\gamma$-IR-mediated CD23 expression. In contrast, Ras, which potentiates $\gamma$-IR-induced NF-${\kappa}B$ activity in these cells, further augmented the $\gamma$-IR- or IL-4-induced CD23 levels, The induction of NF-${\kappa}B$ activation and the subsequent up-regulation of CD23 expression by $\gamma$-IR were also observed in monocytic cells. These results suggest that $\gamma$-IR, at specific dosages, can modulate immune cell differentiation through the activation of NF-${\kappa}B$, and this potentially affects the immune inflammatory response that is mediated by cytokines.

• The Journal of Internal Korean Medicine
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• v.44 no.3
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• pp.294-312
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• 2023

Objective: The purpose of this study is to evaluate the effects of GGX on an ovalbumin (OVA)-induced asthma mice model. Methods: Balb/c mice were challenged with OVA and then treated with three concentrations of GGX (100, 200, and 400 mg/kg). After sacrifice, the bronchoalveolar lavage fluid (BALF) or lungs of the mice were analyzed by fluorescence-activated cell sorting, ELISA, real-time PCR, H&E, Masson's trichrome, PAS and AB-PAS staining, and immunohistofluorescence staining. Results: GGX significantly inhibited the increase of total cells, immune cells (lymphocyte, neutrophils, macrophage, CD4+, CD8+, CD4+CD69+, CD62L-CD44high+, Gr-1+SiglecF-), and the expression of cytokines (IL-4, IL-5, IL-13, IFN-γ) in BALF. It also significantly inhibited the increase of total cells, immune cells (lymphocyte, neutrophils, eosinophil/macrophage, CD3+, CD19+, CD3+CD193+, CD4+, CD8+, CD4+CD69+, CD62L-CD44high+, and Gr-1+SiglecF-), and the expression of IL-13, TARC, and MCP-1 in lung tissue. GGX decreased the severity of histological lung injury and the expressions of STAT3 and GATA3. Conclusion: This study suggests the probability of using GGX for the treatment of asthma by inhibiting inflammatory immune response.

• Parasites, Hosts and Diseases
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• v.54 no.2
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• pp.123-132
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• 2016

Trichomonas vaginalis causes the most prevalent sexually transmitted infection worldwide. Trichomonads have been detected in prostatic tissues from prostatitis, benign prostatic hyperplasia (BPH), and prostate cancer. Chronic prostatic inflammation is known as a risk factor for prostate enlargement, benign prostatic hyperplasia symptoms, and acute urinary retention. Our aim was to investigate whether T. vaginalis could induce inflammatory responses in cells of a benign prostatic hyperplasia epithelial cell line (BPH-1). When BPH-1 cells were infected with T. vaginalis, the protein and mRNA of inflammatory cytokines, such as CXCL8, CCL2, IL-$1{\beta}$, and IL-6, were increased. The activities of TLR4, ROS, MAPK, JAK2/STAT3, and NF-${\kappa}B$ were also increased, whereas inhibitors of ROS, MAPK, PI3K, NF-${\kappa}B$, and anti-TLR4 antibody decreased the production of the 4 cytokines although the extent of inhibition differed. However, a JAK2 inhibitor inhibited only IL-6 production. Culture supernatants of the BPH-1 cells that had been incubated with live T. vaginalis (trichomonad-conditioned medium, TCM) contained the 4 cytokines and induced the migration of human monocytes (THP-1 cells) and mast cells (HMC-1 cells). TCM conditioned by BPH-1 cells pretreated with NF-${\kappa}B$ inhibitor showed decreased levels of cytokines and induced less migration. Therefore, it is suggested that these cytokines are involved in migration of inflammatory cells. These results suggest that T. vaginalis infection of BPH patients may cause inflammation, which may induce lower urinary tract symptoms (LUTS).

• The Journal of Korean Medicine
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• v.27 no.2 s.66
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• pp.196-210
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• 2006

Objectives : This study aimed to find the curative effect of Gamitongkyutang distillate in mice with allergic rhinitis. Metbods : Forty mice were divided into four groups: the normal group, the control group (allergic rhinitis elicited group), the sample I group (Gamitongkyutang treated group after allergic rhinitis elicitation) and the sample II group (distillate of Gamitongkyutang treated group after allergic rhinitis elicitation). Indexes of AR were investigated such as the histological changes of the nasal mucosa, the changes of eosinophil count, the changes of interleukin-4(IL-4) secretion in the intranasal mucosa, the alteration of inducible nitric oxide synthase(iNOS) mRNA expression and the distribution of the nuclear factor kappa B (NF-kB). ANOVA test was used for statistical analysis (p<0.05). Results : Loss of the cilium and the mucous secretion in the sample I and II groups was rare when compared to the control group. The segment of eosinophil was significantly decreased in the sample I and II groups when compared to the control group (p<0.05). A significant decrease of IL-4 mRNA expression was observed in the sample I and II groups when compared with the control group (p<0.05). Inhibition of iNOS induced by NF-kB p50 in the sample I and II groups was significantly superior to that in the control group (p<0.05). DGT and GT didn't affect AST and ALT. Conclusions : GT was superior to DGT in the IL-4 secretion, eosinophil levels and iNOS production. However, considering the difficulty in taking herbal medicine, the DGT has a meaningful curative effect in mice with allergic rhinitis.

• The Journal of Pediatrics of Korean Medicine
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• v.31 no.3
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• pp.37-45
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• 2017

Objectives Hataedock is an orally-administered herbal extract that used to treat toxic heat and help excrete meconium accumulates in the fetus from pregnancy. This study compared the effects of Hataedock using the extract of Douchi and omega-3 to control inflammation in the colonic mucosa. Methods NC/Nga mice were divided into three groups; Control group (no treatment), FGT group (given the extract of Douchi), and O3T group (given omega-3). The 3-week-old mouse from each group were given the extract of Douchi and omega-3. After 2 weeks, the colonic mucosa tissues of the mice were observed via Immunohistochemical staining for IL-4, IL-13, CD40, $Fc{\varepsilon}RI$, $p-I{\kappa}B$, EGF in the intestinal mucosa. Results In both FGT and O3T group, IL-4, IL-13, STAT-6 levels and CD40-positive reactions were decreased. However, EGF-positive responses were increased. FGT group showed 35% decrease in IL-4, 45% decrease in IL-13, 48% decrease in CD40, 50% decrease in $Fc{\varepsilon}RI$, 54% decrease in $p-I{\kappa}B$, 52% increase in EGF compared to the control group. O3T group also showed 14% decrease in IL-4, 23% decrease in IL-13, 22% decrease in CD40, 29% decrease in $Fc{\varepsilon}RI$, 41% decrease in $p-I{\kappa}B$, 39% increase in EGF compared to the control group. Conclusions Both early administration of Douchi extract and omega-3 were effective in regulating Th2, relieving inflammation and developing colonic mucosal tissues. Hataedock with Douchi extract may be more effective in modulating immune system in colonic mucosa than omega-3.

• Development and Reproduction
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• v.12 no.2
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• pp.159-168
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• 2008

Nanog is a newly identified member of the homeobox family of DNA binding transcription factors that functions to maintain the undifferentiated state of stem cells. However, molecular mechanisms underlying the function of Nanog remain largely unknown. To elucidate the regulatory roles of Nanog involved in maintenance of P19 embryonal carcinoma (EC) stem cells, we transfected three small interfering RNA (siRNA) duplexes targeted against different regions of the Nanog gene into P19 cells. The Nanog siRNA-100 duplexes effectively decreased the expression of Nanog up to 30.7% compared to other two Nanog siRNAs, the Nanog siRNA-400 (67.9 %) and -793 (53.0%). When examined by RT-PCR and real-time PCR, the expression of markers for pluripotency such as Fgf4, Oct3/4, Rex1, Sox1 and Yes was downregulated at 48 h after transfection with Nanog siRNA-100. Furthermore, expression of the ectodermal markers, Fgf5 and Isl1 was reduced by Nanog knockdown. By contrast, the expression of other markers for pluripotency such as Cripto, Sox2 and Zfp57 was not affected by Nanog knockdown at this time. On the other hand, the expression of Lif/Stat3 pathway molecules and of the endoderm markers including Dab2, Gata4, Gata6 and the germ cell nuclear factor was not changed by Nanog knockdown. The results of this study demonstrated that the knockdown of Nanog expression by RNA interference in P19 cells was sufficient to modulate the expression of pluripotent markers involved in the self-renewal of EC stem cells. These results provide the valuable information on potential downstream targets of Nanog and add to our understanding of the function of Nanog in P19 EC stem cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.30 no.1
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• pp.29-42
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• 2017

Objectives : Atopic dermatitis accompanies with severe pruritus and collapse of skin barrier, inflammation. Maifanite could be used as an ointment for skin disease. However, there have been few studies about maifanite uses for atopic dermatitis. We report the anti-inflammatory and promoting skin recovery effects of ionized maifanite on damaged skin barrier with experimentally elicited atopic dermatitis. Methods : Nc/Nga mice were divided into 3 groups: control group(CON), atopic dermatitis elicited group(AE group), ionized maifanite treated group after atopic dermatitis elicitation(MT group). After 5% SDS was applied D. pteronyssinus crude extract also applied for 3 weeks to elicit atopic dermatitis-like skin disease. MT group was treated for 3 weeks with ionized maifanite. Ionized maifanite was applied once a day and voluntarily administrated. AE group and control group were treated with normal saline in the same way. Results : In MT group, skin lesions like eczema were more improved than AE group. p-ERK1/2 positive reaction was reduced in MT group. MMP-9 and substance P positive reaction at dermal papillae was also reduced in MT group. With skin angiogram, capillary vessel decreased in MT group. Also, IL-4 positive reaction cell and STAT-6 positive reaction cell reduced more in MT group than in AE group. $NF-{\kappa}B$ p65 positive reaction cell and iNOS positive reaction cell also declined more in MT group than in AE group. Conclusions : It is supposed that ionized maifanite has anti-inflammatory effects on NC/Nga mice's atopic dermatitis with suppressing IL-4 production and Th2 cell differentiation, and controlling $NF-{\kappa}B$ activation.

• The Journal of Korean Medicine
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• v.39 no.4
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• pp.74-85
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• 2018

Objectives: HTD treatment is a traditional preventive therapy for neonatal inflammatory diseases such as AD. The aim of this study was to investigate the efficacy of HTD treatments for the maintenance and formation of lipid barrier in Dermatophagoides farina-induced obese NC/Nga mice. Methods: 20 mg/kg of CRGR extracts as HTD treatments were orally administered to NC/Nga mice. To induce obesity, high fat diet was served. Dermatophagoides farina extracts was applied on the 4th-6th and 8th-10th weeks to induce AD-like skin lesions in NC/Nga mice. Changes of skin conditions in mice were observed by histochemistry and immunohistochemistry. Results: The results showed that HTD treatments effectively maintained and formed the lipid barrier. In the experimental groups, restorations of Lass2 expression and distributions of filaggrin, involucrin, loricrin, ASM, and LXR means that HTD treatments maintained and generated the lipid barrier. In the dermal papillae, HTD treatments reduced PKC production accompanied by epidermis damage. Furthermore, levels of IL-4, and STAT6 was low. HTD treatment may be effective for preventing inflammation induced by Th2-skewed condition by suppressing the main pathway of Th2 differentiation. Conclusions: HTD treatment alleviated the inflammatory damage in the skin tissues of the NC/Nga mice by maintaining the lipid barrier and suppressing Th2 differentiation.