• Food Science and Biotechnology
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• v.18 no.5
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• pp.1063-1070
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• 2009

Dangyuja (Citrus grandis Osbeck) is a native plant growing only on Jeju Island in Korea. In this study, antiinflammatory effect of dangyuja leaves on a murine macrophage cell line was investigated. RAW 264.7 murine macrophage cells were stimulated with lipopolysaccharide (LPS, $1{\mu}g/mL$) to induce expression of pro-inflammatory markers [interleukin (IL)-6 and inducible nitric oxide synthase (iNOS)]. The crude extract (80% MeOH Ex.) and solvent fractions (hexane, $CHCl_3$, EtOAc, BuOH, and $H_2O$ Ex.) were obtained from dangyuja leaves. The $CHCl_3$ fraction inhibited the nitric oxide (NO) and IL-6 production in a dose-dependent manner. Also, the $CHCl_3$ fraction inhibited mRNA expression and protein levels of iNOS in a dose-dependent manner. Furthermore, the $CHCl_3$ fraction inhibited LPS-induced nuclear factor (NF)-${\kappa}B$ activation and phosphorylation of mitogen-activated protein kinases (MAPKs: ERK, JNK, and p38). These results suggest that dangyuja leaves may inhibit LPS-induced production of inflammatory markers by blocking NF-${\kappa}B$ and MAPKs signaling in RAW 264.7 cells.

• Korean Journal of Ecology and Environment
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• v.50 no.1
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• pp.137-147
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• 2017

The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.627-636
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• 2011

A commercially important alkaline protease, produced by Bacillus sp. RRM1 isolated from the red seaweed Kappaphycus alvarezii (Doty) Doty ex Silva, was first recognized and characterized in the present study. Identification of the isolated bacterium was done using both biochemical characterization as well as 16S rRNA gene sequencing. The bacterial strain, Bacillus sp. RRM1, produced a high level of protease using easily available, inexpensive agricultural residues solid-state fermentation (SSF). Among them, wheat bran was found to be the best substrate. Influences of process parameters such as moistening agents, moisture level, temperature, inoculum concentration, and co-carbon and co-nitrogen sources on the fermentation were also evaluated. Under optimized conditions, maximum protease production (i.e., 2081 U/g) was obtained from wheat bran, which is about 2-fold greater than the initial conditions. The protease enzyme was stable over a temperature range of 30-$60^{\circ}C$ and pH 6-12, with maximum activity at $50^{\circ}C$ and pH 9.0. Whereas the metal ions $Na^+$, $Ca^{2+}$, and $K^+$ enhanced the activity of the enzyme, others such as $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, $Co^{2+}$, and $Zn^{2+}$ had rendered negative effects. The activity of the enzyme was inhibited by EDTA and enhanced by $Cu^{2+}$ ions, thus indicating the nature of the enzyme as a metalloprotease. The enzyme showed extreme stability and activity even in the presence of detergents, surfactants, and organic solvents. Moreover, the present findings opened new vistas in the utilization of wheat bran, a cheap, abundantly available, and effective waste as a substrate for SSF.

• Journal of Acupuncture Research
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• v.24 no.4
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• pp.157-166
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• 2007

Objective : The aim of this study was to investigate anti-arthritic effects of Honghwa-ja herbal acupuncture extract through inhibitory MIF activation. Methods: After Rheumatoid arthritis(RA) knee joint was induced by lipopolysaccharide(LPS) in vivo, Honghwa-ja herbal acupuncture solution was applied on $ST_{35}$(犢鼻) and $EX-LE_{202}$(內膝眼) coresponding human body. To research the effect on the expression of MIF, COX-2, MMP-9 mRNA, RT-PCR was performed on LPS-stimulated Raw 264.7 cells. Results: In the Honghwa-ja herbal acupuncture solution treated Raw 264.7 cell, the mRNA expression of cytokines, RA related inflammation factors, such as the MIF, COX-2, and MMP-9 reduced concentration dependently. Positive reaction of RA-related cytokines MIF, $IL-6R-{\alpha}$, STAT3, COX-2, MMP-9 was reduced. Conclusion : Honghwa-ja herbal acupuncture extract has significant protecting ability against acute progressive RA by inhibiting the production of MIF, as a top in cytokines related to inflammation.

• Journal of Genetic Medicine
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• v.20 no.1
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• pp.6-14
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• 2023

Fabry disease (FD), a rare X-linked lysosomal storage disorder, is caused by mutations in the α-galactosidase A gene gene encoding α-galactosidase A (α-Gal A). The functional deficiency of α-Gal A results in progressive accumulation of neutral glycosphingolipids, causing multi-organ damages including cardiac, renal, cerebrovascular systems. The current treatment is comprised of enzyme replacement therapy (ERT), oral pharmacological chaperone therapy and adjunctive supportive therapy. ERT has been introduced 20 years ago, changing the outcome of FD patients with proven effectiveness. However, FD patients have many unmet needs. ERT needs a life-long intravenous therapy, inefficient bio-distribution, and generation of anti-drug antibodies. Migalastat, a pharmacological chaperone, augmenting α-Gal A enzyme activity only in patients with mutations amenable to the therapy, is now available for clinical practice. Furthermore, these therapies should be initiated before the organ damage becomes irreversible. Development of novel drugs aim at improving the clinical effectiveness and convenience of therapy. Clinical trial of next generation ERT is underway. Polyethylene glycolylated enzyme has a longer half-life and potentially reduced antigenicity, compared with standard preparations with longer dosing interval. Moss-derived enzyme has a higher affinity for mannose receptors, and seems to have more efficient access to podocytes of kidney which is relatively resistant to reach by conventional ERT. Substrate reduction therapy is currently under clinical trial. Gene therapy has now been started in several clinical trials using in vivo and ex vivo technologies. Early results are emerging. Other strategic approaches at preclinical research level are stem cell-based therapy with genome editing and systemic mRNA therapy.

• BMB Reports
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• v.49 no.3
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• pp.191-196
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• 2016

Vascular endothelial growth factor (VEGF) is a key mediator of angiogenesis and critical for normal embryonic development and repair of pathophysiological conditions in adults. Although phospholipase D (PLD) activity has been implicated in angiogenic processes, its role in VEGF signaling during angiogenesis in mammals is unclear. Here, we found that silencing of PLD2 by siRNA blocked VEGF-mediated signaling in immortalized human umbilical vein endothelial cells (iHUVECs). Also, VEGF-induced endothelial cell survival, proliferation, migration, and tube formation were inhibited by PLD2 silencing. Furthermore, while Pld2-knockout mice exhibited normal development, loss of PLD2 inhibited VEGF-mediated ex vivo angiogenesis. These findings suggest that PLD2 functions as a key mediator in the VEGF-mediated angiogenic functions of endothelial cells.

• IMMUNE NETWORK
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• v.21 no.1
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• pp.3.1-3.16
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• 2021

Coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading worldwide since its outbreak in December 2019, and World Health Organization declared it as a pandemic on March 11, 2020. SARS-CoV-2 is highly contagious and is transmitted through airway epithelial cells as the first gateway. SARS-CoV-2 is detected by nasopharyngeal or oropharyngeal swab samples, and the viral load is significantly high in the upper respiratory tract. The host cellular receptors in airway epithelial cells, including angiotensin-converting enzyme 2 and transmembrane serine protease 2, have been identified by single-cell RNA sequencing or immunostaining. The expression levels of these molecules vary by type, function, and location of airway epithelial cells, such as ciliated cells, secretory cells, olfactory epithelial cells, and alveolar epithelial cells, as well as differ from host to host depending on age, sex, or comorbid diseases. Infected airway epithelial cells by SARS-CoV-2 in ex vivo experiments produce chemokines and cytokines to recruit inflammatory cells to target organs. Same as other viral infections, IFN signaling is a critical pathway for host defense. Various studies are underway to confirm the pathophysiological mechanisms of SARS-CoV-2 infection. Herein, we review cellular entry, host-viral interactions, immune responses to SARS-CoV-2 in airway epithelial cells. We also discuss therapeutic options related to epithelial immune reactions to SARS-CoV-2.

• Journal of Acupuncture Research
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• v.23 no.6
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• pp.117-132
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• 2006

Objectives : The purpose of this study is to investigate the effect of Ulmus davidiana Planch herbal acupuncture solution in LPS-stimulated RAW 264.7 cells and mouse knee joints, perfom1ed several experimental items: those are MIF mRNA, MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS mRNA, iNOS, NO, synovial hyperplasia, angiogenesis and fibrosis. Methods : In order to observe mRAN expression of MIF and iNOS in LPS-stimulated RAW 264.7 cells, RT-PCR was used. NO production in LPS-stimulated RAW 264.7 cells was measured by nitrite assay. All the female BALB/c mice were bred and maintained in pathogen-free mouse colonies and were 6 weeks of age on beginning of the experiment. The experimental model of RA was induced by injection of $50{\mu}g/kg$ LPS. Ulmus davidiana Planch herbal acupuncture solution was injected into either S 35 (犢鼻) or EX-LE 202 (內膝眼) of mice in turn daily for 19 days. Immunohistochemical staining was carried out to assess $TNF-{\alpha}$, $NF-{\kappa}B$ p65 and iNOS expression in synovial membrane. Synovial hyperplasia, angiogenesis and fibrosis in synovial membrane was observed with a microscope. Results : 1. Ulmus davidiana Planch herbal acupuncture solution inhibited mRNA expression of MIF and iNOS in dependence on a density of it in LPS-stimulated RAW 264.7 cells. 2. Ulmus davidiana Planch herbal acupuncture solution decreased synovial hyperplasia, angiogenesis and fibrosis in LPS-stimulated mouse knee joints. 3. Ulmus davidiana Planch herbal acupuncture solution curtailed production of MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS in LPS-stimulated mouse knee joints. Conclusion : On the basis of these results, It was shown that Ulmus davidiana Planch herbal acupuncture solution is significantly able to inhibit the production of MIF as a top in cytokines related to inflammatory or irrlll1une responses. Our results may provide that Ulmus davidiana Planch herbal acupuncture solution has beneficial effect in not only RA but other inflammatory or immune deases.

• Journal of Acupuncture Research
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• v.24 no.4
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• pp.167-181
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• 2007

Objectives: To evaluate expression of MIF mRNA, MIF, $TNF-{\alpha}$, $IL-6R-{\alpha}$, STAT-3, $NF-{\kappa}B$ p65, COX-2 and iNOS, MMP-9mRNA after injecting deer antler herbal acupuncture solution in a LPS rat model. Methods: The experiment was divided in category of the control group, RA group, and NA group. RA was induced in the mice via injecting 300ug/kg LPS. The deer antler herbal acupuncture solution 50ug/kg was applied on $ST_{35}$(犢鼻) and EX-LE201(內膝眼) for 19days from $3^{rd}$ day of RA inducement. Results: 1. In the deer antler herbal acupuncture solution treated RAW 264.7cell, the mRNA expression of cytokines, RA related inflammation factors, such as the MIF, COX- 2, iNOS, and MMP-g reduced concentration dependently. 2. In the deer antler herbal acupuncture treated mice's synovial membrane, decrease in the cell replication of synovial joint cells, regeneration of blood vessel, fibrosis and fibroblastic cells expansion were observed. 3. Positive reaction of RA-related cytokines MIF, $TNF-{\alpha}$, $IL-6R-{\alpha}$, STAT3, COX-2, iNOS, $NF-\;{\kappa}B$ p65, MMP-9 was reduced. Conclusion : On the basis of the results, it was concluded that deer antler herbal acupuncture extract has significant protecting ability against acute progressive RA by inhibiting the production of MIF, as a top in cytokines related to inflammation.

• Korean Journal of Medicinal Crop Science
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• v.26 no.1
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• pp.42-54
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• 2018

Background: Allergic diseases like such as atopic dermatitis, asthma, and rhinitis have recently increased both domestically and globally. The present study was undertaken to select candidates with anti-allergic activity from plant resources. Methods and Results: Fifty-six plant extracts at $20{\mu}g/m{\ell}$ were screened against ${\beta}$-hexosaminidase production and interleukin (IL)-4 release in degranulated rat basophilic leukemia (RBL)-2H3 cells. The anti-allergy activity of three plant extracts selected from the preliminary screening experiment, Polygonatum sibiricum F. Delaroche (root), Pyrus pytifolia var. culta (Makino) Nakai (leaf), and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud (root) were measured at concentrations of $2-250{\mu}g/m{\ell}$ in three cell lines as RBL-2H3, HaCaT and Jurkcat T cells. The assay showed the root extract of R. glutinosa to have an inhibitory activity of 4.2% - 28.6% on ${\beta}$-hexosaminidase production from IgE-sensitized RBL-2H3 cells. Each extract of P. sibiricum and R. glutinosa reduced IL-4 release in IgE-sensitized RBL-2H3 cells, respectively. The leaf extract of P. pyrifolia var. culta showed a significantly potent suppressive effect of 10.2% - 74.7% on the mRNA expression of tumor necrosis factor (TNF)-${\alpha}$ in HaCaT cells sensitized with TNF-a and INF-g, and showed inhibitory effect of -8.6% - 90.9% on the mRNA expression of IL-2 in Jurkat T cells sensitized with PMA and A23187. Conclusions: The results showed that the root of R. glutinosa and leaf of P. pyrifolia var. culta could be useful candidates as antiallergy materials.