• The Korean Journal of Physiology and Pharmacology
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• v.22 no.3
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• pp.349-360
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• 2018

Autophagy has been studied as a therapeutic strategy for cardiovascular diseases. However, insufficient studies have been reported concerning the influence of vascular smooth muscle cells (VSMCs) through autophagy regulation. The aim of the present study was to determine the effects of VSMCs on the regulation of autophagy under in vitro conditions similar to vascular status of the equipped micro-tubule target agent-eluting stent and increased release of platelet-derived growth factor-BB (PDGF-BB). Cell viability and proliferation were measured using MTT and cell counting assays. Immunofluorescence using an $anti-{\alpha}-tubulin$ antibody was performed to determine microtubule dynamic formation. Cell apoptosis was measured by cleavage of caspase-3 using western blot analysis, and by nuclear fragmentation using a fluorescence assay. Autophagy activity was assessed by microtubule-associated protein light chain 3-II (LC-II) using western blot analysis. Levels of intracellular reactive oxygen species (ROS) were measured using $H_2DCFDA$. The proliferation and viability of VSMCs were inhibited by microtubule regulation. Additionally, microtubule-regulated and PDGF-BB-stimulated VSMCs increased the cleavage of caspase-3 more than only the microtubule-regulated condition, similar to that of LC3-II, implying autophagy. Inhibitory autophagy of microtubule-regulated and PDGF-BB-stimulated VSMCs resulted in low viability. However, enhancement of autophagy maintained survival through the reduction of ROS. These results suggest that the apoptosis of conditioned VSMCs is decreased by the blocking generation of ROS via the promotion of autophagy, and proliferation is also inhibited. Thus, promoting autophagy as a therapeutic target for vascular restenosis and atherosclerosis may be a good strategy.

• Microbiology and Biotechnology Letters
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• v.50 no.1
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• pp.22-30
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• 2022

Haliotis discus hannai called abalone, is the valuable marine mollusks and the by-products of abalone processing are viscera. Brownish abalone male viscera (AMV), which have not been reported as having anti-inflammatory effects, was extracted with acetone and fractionated by different six acetone/hexane ratios (0, 10, 20, 30, 40, and 100%) using a silica column via in vitro ABTS and DPPH radical and nitric oxide (NO) production assay-guided fractionation. Among the fractions, the acetone/hexane ratio 40%, A40 exhibited the most potent radical scavenging activities and inhibition of lipopolysaccharide (LPS)-induced NO production without cytotoxicity. A40 inhibited LPS-induced intracellular reactive oxygen species (ROS) production in a dose-dependent manner. Western blot analysis revealed that A40 down-regulated the activation of NF-κB, MAPK (ERK 1/2, p-38, and JNK), and inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Moreover, this fraction inhibited the generation of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. These results suggested that AMV containing A40 with anti-inflammatory and anti-oxidantive effects, is the effective therapeutic and functional material for treating inflammatory disorders.

• Korean Journal of Plant Resources
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• v.22 no.3
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• pp.209-214
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• 2009

To facilitate the introgression of F. esculentum into the traits of F. homotropicum, several accessions of the hybrids between these two species were pollinated with F. esculentum as the recurrent parent. The embryo in vitro rescue was performed to increase the recovery of backcross progenies. The $F_{2}$ generation was more amenable than $F_{1}$ hybrids to produce backcross progenies. The $F_{1}$ hybrids were backcrossed twice with common buckwheat (pin-type F. esculentum) (recurrent backcrossing). Also, alternate backcrosses with common buckwheat and F. homotropicum (congruity backcrossing) were carried out. Pollen tube growth of BC$F_{1}$ ${\times}$ F. esculentum (thrum) and F. homotropicum ${\times}$ BC$F_{1}$ was the disturbed penetration exceeded for all initial interspecific hybrids, and its requirement was proportionally lower when the common buckwheat was used as the recurrent parent and as the last parent of congruity hybrids. Effects of both common buckwheat and F. homotropicum on seed success rate for hybridization were observed. Growth of hybrid embryos before rescue, regeneration of mature hybrids all increased recurrent and congruity backcrosses and inter-crosses between $F_{1}$ plants and selected fertile plants of the second congruity backcrosses.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.92-103
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• 2021

Lactobacillus casei, one of the most widely used probiotics, has been reported to alleviate multiple diseases. However, the effects of this species on intestinal diseases are strain-specific. Here, we aimed to screen L. casei strains with inflammatory bowel disease (IBD)-alleviating effects based on in vitro physiological characteristics. Therefore, the physiological characteristics of 29 L. casei strains were determined, including gastrointestinal transit tolerance, oligosaccharide fermentation, HT-29 cell adhesion, generation time, exopolysaccharide production, acetic acid production, and conjugated linoleic acid synthesis. The effects of five candidate strains on mice with induced colitis were also evaluated. The results showed that among all tested L. casei strains, only Lactobacillus casei M2S01 effectively relieved colitis. This strain recovered body weight, restored disease activity index score, and promoted anti-inflammatory cytokine expression. Gut microbiota sequencing showed that L. casei M2S01 restored a healthy gut microbiome composition. The western blotting showed that the alleviating effects of L. casei M2S01 on IBD were related to the inhibition of the NF-κB pathway. A good gastrointestinal tolerance ability may be one of the prerequisites for the IBD-alleviating effects of L. casei. Our results verified the efficacy of L. casei in alleviating IBD and lay the foundation for the rapid screening of L. casei strain with IBD-alleviating effects.

• Tuberculosis and Respiratory Diseases
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• v.48 no.6
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• pp.887-897
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• 2000

Background : Since the exact pathogenesis of sepsis-induced ARDS has not been elucidated, the mechanisms of enhanced neutrophilic respiratory burst were probed in endotoxin primed neutrophils associated with the roles of phospholipase A2(PLA2), platelet activating factor(PAF) and apoptosis. Methods : In isolated fresh human neutrophils, effects of the inhibition of PLA2 and PAF on the apoptosis were examined by the method of Annexin-FITC/dual PIflow cytometry. The roles of PLA2 and PAF on the neutrophilic respiratory burst were also examined by measuring oxidant generation in cytochrome-c reduction assay. Activities of the PLA2 and lysoPAF acetyltransferase (lysoPAF AT) of the neutrophils were determined to understand the effect of endotoxin on these enzymatic activities which may be related to the neutrophilic respiratory burst and apoptosis. In addition, the role roles of PLA2 and PAF in neutrophilic adhesion to bovine endothelial cells were examined in vitro by neutrophil adhesion assay. To investigate the effect of oxidants on pulmonary surfactant, cytochemical ultrastructural microscopy was performed. To inhibit PLA2 and PAF, non-specific PLA2 inhibitor mepacrine (100 nM) and WEB 2086 (100 nM) or ketotifen fumarate (10 ${\mu}g$/ml) were used respectively in all in vitro experimental sets. WEB 2086 is PAF receptor antagonist, and ketotifen fumarate is a lyso PAF AT inhibitor. Results: The mapacrine treatment, provided and the endotoxin (ETX) treatment, resulted in increased apoptosis of neutrophils (p<0.001) while treatments of WEB 2086 and ketotifen did not. The inhibition of PLA2 and PAF decreased (p<0.001) production of oxidants from PMA-stimulated neutrophils. While endotoxin increased the PLA2 activity of neutrophils (p<0.01), mepacrine supressed (p<0.001) the activity, provided after treatment of ETX. The lyso PAF actyltransferase activity (lyso PAF AT) increased (p<0.01) after treatment of ETX. In contrast, mepacrine, WEB 2086 and ketotifen showed a tendency of decreasing the activity after treatment of ETX. The treatment of ETX incresed (p<0.001) neutrophil adhesion to endothelial cells, which was reversed by inhibition of PLA2 and PAF (p<0.01). The binding of oxidants to pu1monary surfactant was identified histologically. Conclusions : The enhanced neutrophilic respiratory burst by ETX plays a pivotal role in the pathogenesis of ARDS in terms of oxidayive oxidative stress. Increased production of oxidants from neutrophils is mediated by the activations of PLA2 and lyso PAF AT.

• Applied Biological Chemistry
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• v.49 no.1
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• pp.21-24
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• 2006

A series of cytotoxic activities $(ED_{50})$ in vitro against six human cancers (lung cancer, uterine cancer, skin cancer, brain cancer, colon cancer and adenocarcinoma) and their seventeen cell lines of 3,6-bis(2'-pyridyl)pyridazine, 1, 3,6-bis-(6'-methyl-2'-pyridyl)pyridazine, 2 and their transition metal, Ni(II), Cu(II) and Zn(II) complexes, $3{\sim}6$ were measured. Particularly, the results revealed that the cytotoxic activities against the brain cancer cell line (SNB-19) and the colon cancer cell line (SW62) of bis- [3,6-bis-(6'-methyl-2'-pyridyl)pyridazine-$k^2N^2,N^3$]chlorocopper(II)perchlorate, 4 were shown to be higher than that of the first generation anticancer agent, Cis-platin.

• The Journal of Korean Academy of Prosthodontics
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• v.51 no.1
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• pp.11-19
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• 2013

Purpose: The aim of this study was to evaluate the heat generation in bone in vitro during the guided flapless drilling procedure and the effect of drilling methods on the heat generation. Materials and methods: A model that has missing the first and second mandibular molars bilaterally was used. In group A, classical flap implant surgery was performed. In group B, flapless implant surgery using surgical guide was performed. In group C, flapless implant surgery using surgical guide without up-and-down pumping motion was performed. Temperature was measured with k-type thermocouple and a real-time digital thermometer. The thermocouples were placed at 0.5 mm away from the osteotomy area at the depths of 3 mm and 6 mm. The measured values were evaluated with independent t-test. Results: The mean temperature generated was $27.2^{\circ}C$ ($SD{\pm}2.1^{\circ}C$) and $27.5^{\circ}C$ ($SD{\pm}2.3^{\circ}C$) for groups A and B, respectively. These differences were not statistically significant. In group C, the mean temperature was $37.0^{\circ}C$ ($SD{\pm}3.4^{\circ}C$). There were statistically significant differences between groups B and C with respect to the mean temperature. Conclusion: These findings suggest that guided flapless drilling with up-and-down pumping motion may not significantly increase the bone temperature.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.269-275
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• 1996

Biosynthesis and secretion of anterior pituitary hormones are under the control of specific hypothalamic stimulatory and inhibitory factors. Among them, Growth Hormone Releasing Hormone (GHRH) is the major stimulator of pituitary somatotrophs activating GH gene expression and secretion. Human GHRH is a polypeptide of 44 amino acids initially isolated from pancreatic tumors, and the gene for the hypothalamic form of GHRH is organized into 5 exons spanning over 10 kilobases (kb) on genomic DNA and encodes a messenger RNA of 700-750 nucleotides. Several neuropeptides classically associated with the hypothalamus have been found in the extrahypothalamic regions, suggesting the existence of novel sources, targets and functions. GHRH-like immunoreactivity has been found in several peripheral sites, including placenta, testis, and ovary, indicating that GHRH may also have regulatory roles in peripheral reproductive organs. Furthermore, higher molecular weight forms of the GHRH transcripts were identified from these organs (1.75 kb in testis; 1.75 and >3 kb in ovary). These tissue-specific expression of GHRH gene suggest the existence of unique regulatory mechanism of GHRH expression and function in these organs. In fact, placenta-specific and testis-specific promoters for GHRH transcripts which are located in about 10 kb upstream region of hypothalamic promoter were reported. The use of unique promoters in extrahypothalamic sites could be refered in a different control of GHRH gene and different functions of the translated products in these tissues. Somatotrophs and lactotrophs have been thought to be derived from a common bipotential progenitor, the somatolactotrophs, which give origins to either phenotypes. Although the precise mechanism responsible for the lactotroph differentiation in the anterior pituitary gland has not been yet clalified, there are several candidators for the generation of lactotrophs. In human, the presence of GHRH peptides with different size from authentic hypothalamic form in the normal anterior pituitary and several types of adenoma were demonstrated. Recently our group found the existence of immunoreactive GHRH and its transcript from the normal rat anterior pituitary (gonadotroph> somatotroph> lactotroph), and the GHRH treatment evoked the increased proliferation rate of anterior pituitary cells in vitro. The transgenic mouse models clearly shown that GHRH or NGF overexpression by anterior pituitary cells induced development of pituitary hyperplasia and adenomas particularly GH-oma and prolactinoma. Taken together, we hypothesize that the pituitary GHRH could serve not only as a modulator of hormone secretion but as a paracrine or autocrine regulator of anterior pituitary cell proliferation and differentiation. Interestingly enough, the expression of Pit-1 homeobox gene (the POU class transcription factor) was confined to somatotrophs, lactotrophs and somatolactotrophs in which GHRH receptors are expressed commonly. Concerning the mechanism of somatolactotroph and lactotroph differentiation in the anterior pituitary, we have focused following two possibilities; (1) changes in the relative levels or interactions of both hypothalamic and intrapituitary factors such as dopamine, VIP, somatostatin, NGF and GHRH; (2) alterations of GHRH-GHRH receptor signaling and Pit-1 activity may be the cause of lactotroph differentiation or pituitary hyperplasia and adenoma formation. Extensive further studies will be necessary to solve these complicated questions.

• Journal of Ginseng Research
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• v.46 no.3
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• pp.418-425
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• 2022

Background: Sorafenib is effective in treating hepatoma, but most patients develop resistance to it. STAT3 signaling has been implicated in sorafenib resistance. Artesunate (ART) and 20(R)-ginsenoside Rg3 (Rg3) have anti-hepatoma effects and can inhibit STAT3 signaling in cancer cells. This study aimed to evaluate the effects of Rg3 in combination with ART (Rg3-plus-ART) in overcoming sorafenib resistance, and to examine the involvement of STAT3 signaling in these effects. Methods: Sorafenib-resistant HepG2 cells (HepG2-SR) were used to evaluate the in vitro anti-hepatoma effects of Rg3-plus-ART. A HepG2-SR hepatoma-bearing BALB/c-nu/nu mouse model was used to assess the in vivo anti-hepatoma effects of Rg3-plus-ART. CCK-8 assays and Annexin V-FITC/PI double staining were used to examine cell proliferation and apoptosis, respectively. Immunoblotting was employed to examine protein levels. ROS generation was examined by measuring DCF-DA fluorescence. Results: Rg3-plus-ART synergistically reduced viability of, and evoked apoptosis in HepG2-SR cells, and suppressed HepG2-SR tumor growth in mice. Mechanistic studies revealed that Rg3-plus-ART inhibited activation/phosphorylation of Src and STAT3 in HepG2-SR cultures and tumors. The combination also decreased the STAT3 nuclear level and induced ROS production in HepG2-SR cultures. Furthermore, overactivation of STAT3 or removal of ROS diminished the anti-proliferative effects of Rg3-plus-ART, and removal of ROS diminished Rg3-plus-ART's inhibitory effects on STAT3 activation in HepG2-SR cells. Conclusions: Rg3-plus-ART overcomes sorafenib resistance in experimental models, and inhibition of Src/STAT3 signaling and modulation of ROS/STAT3 signaling contribute to the underlying mechanisms. This study provides a pharmacological basis for developing Rg3-plus-ART into a novel modality for treating sorafenib-resistant hepatoma.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.37-37
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• 2021

Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L^-1, and IBA 1mg·L^-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.