• Korean Journal of Environmental Biology
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• v.36 no.4
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• pp.498-506
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• 2018

The purpose of this study is to examine the restorative effect of Semisulcospira libertina extract, on damaged liver cells induced by D-galactosamine in rats. Treatment of damaged liver cells with S. libertina extract significantly reduced local fatty degeneration, and inflammatory cell necrosis, to levels similar with the undamaged control group. In addition, S. libertina extracts were found to reduce plasma levels of liver damage indicator enzymes, such as AST, ALT, LDH and ALP, to control levels. It also reduced lipid peroxides, and lipid contents within damaged liver tissues. This suggests that S. libertina extract has a restorative effect on liver cells, thus reducing release of damage-associated liver enzymes, and oxidative degradation of lipids. Also, S. libertina extracts were found to be involved in recovery of damaged cells from inflammatory response by suppressing expression of $TNF-{\alpha}$, which leads to tissue injury and necrosis, whereas inducing expression of HO-1 that protects cells during inflammation. Thus, S. libertina extract restores liver tissue from necrosis and fibrosis, as well modulates expression of inflammation-related genes against liver damage. Our findings suggest that S. libertina extract is an effective medicinal resource, for improving and recovering liver cells from hepatic injury.

• Clinical and Experimental Pediatrics
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• v.61 no.12
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• pp.403-406
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• 2018

Floating-Harbor syndrome is a rare autosomal dominant genetic disorder associated with SRCAP mutation. To date, approximately 50 cases of Floating-Harbor syndrome have been reported, but none have been reported in Korea yet. Floating-Harbor syndrome is characterized by delayed bony maturation, unique facial features, and language impairment. Here, we present a 6-year-old boy with a triangular face, deep-set protruding eyes, low-set ears, wide nose with narrow nasal bridge, short philtrum, long thin lips, clinodactyly, and developmental delay that was transferred to our pediatric clinic for genetic evaluation. He showed progressive delay in the area of language and cognition-adaption as he grew. He had previously undergone chromosomal analysis at another hospital due to his language delay, but his karyotype was normal. We performed targeted exome sequencing, considering several syndromes with similar phenotypes. Library preparation was performed with the TruSight One sequencing panel, which enriches the sample for about 4,800 genes of clinical relevance. Massively parallel sequencing was conducted with NextSeq. An identified variant was confirmed by Sanger sequencing of the patient and his parents. Finally, the patient was confirmed as the first Korean case of Floating-Harbor syndrome with a novel SRCAP (Snf2 related CREBBP activator protein) mutation (c.7732dupT, p.Ser2578Phefs*6), resulting in early termination of the protein; it was not found in either of his healthy parents or a control population. To our knowledge, this is the first study to describe a boy with Floating-Harbor syndrome with a novel SRCAP mutation diagnosed by targeted exome sequencing in Korea.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.465-472
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• 2019

Candida albicans is an opportunistic human pathogen that causes infections. Candidiasis is often related to antifungal resistance because the pathogen has the ability to form biofilms. In a previous study, we found that the Salvia miltiorriza ethanol extract demonstrated anticandidal activity by altering membrane permeability and inhibiting the cell wall synthesis in C. albicans. Our results here demonstrate that $78{\mu}g/ml$ of the S. miltiorriza extract significantly diminished the early stage biofilms formed by 10 clinical C. albicans isolates by 51.3%; this was analyzed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) reduction assay. The effect of the S. miltiorrhiza extract on the adhesion of C. albicans cells to polystyrene plates and germ tube formation was examined via microscopic investigation. Although the density of the adhered cells was remarkably reduced up on incubation with $39{\mu}g/ml$ S. miltiorrhiza extract, germ tube formation by C. albicans was rarely affected. Quantitative real-time PCR analysis showed that the S. miltiorrhiza extract downregulated the expression of C. albicans hypha-specific genes, EAP1 by 34.7% (p < 0.001), ALS1 by 45.0% (p < 0.001), ALS3 by 48.1% (p < 0.001), and ECE1 by 21.3% (p = 0.006), respectively. Our data suggest that the S. miltiorrhiza ethanol extract significantly inhibited the early stage of biofilm formation by C. albicans by interfering with cell adhesion, by downregulating EAP1, ALS1 and ALS3, and presumably by modifying the cell wall and membrane structure.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.9
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• pp.333-340
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• 2019

This study investigated the effects of goat blood serum (gBS), goat follicular fluid (gFF) and gonadotropin (FSH) on the in vitro maturation, fertilization and development of Korean native black goat oocytes. Our results indicate that the gBS combined with FSH treated group showed significantly higher maturation rate than the other groups. Furthermore, blastocyst formation rate was significantly increased in all treated groups, and gBS and gFF combined with FSH treated groups were higher than other groups. However, gene expression levels of BMP15 and GDF9 in COC, both oocyte maturation related genes, remained unaffected after 24 h maturation. The results of the present study indicate that supplementation of the maturation medium with gBS, gFF and FSH is efficacious in improving the in vitro maturation, fertilization and development of Korea native black goat oocytes.

• Journal of Life Science
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• v.29 no.10
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• pp.1144-1151
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• 2019

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease associated with various metabolic syndromes, such as obesity, dyslipidemia, hypertension, and diabetes. Cudrania tricuspidata is a medicinal plant distributed widely in Asia and has been used in clinical practice to treat various diseases. The aim of this study is to determine the lipid-lowering effects of C. tricuspidata fruit extract (CTE) using a cell model induced by free fatty acids (FFAs). HepG2 cells were exposed to 1mM FFAs (palmitic acid:oleic acid = 2:1) for 24 hr to simulate the conditions of NAFLD in vitro. CTE attenuated the increases of lipid accumulation, intracellular triglyceride, and cholesterol content and inhibited 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) activity in the HepG2 cells in a dose-dependent manner. Also, CTE inhibited the protein expression of lipogenesis-related genes, such as sterol regulatory element-binding protein-1/-2 (SREBP-1/-2), fatty acid synthase (FAS), and stearoyl CoA desaturase-1 (SCD-1) in FFAs-induced lipid accumulation in HepG2 cells. In addition, CTE-induced adenosine monophosphate-activated protein kinase (AMPK) phosphorylation in HepG2 cells. These results suggest that CTE attenuates hepatic lipid accumulation by inhibiting lipogenesis through the modulation of the AMPK signaling pathway on FFAs-induced lipogenesis in HepG2 cells and may potentially prevent NAFLD.

• Journal of Life Science
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• v.29 no.10
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• pp.1152-1158
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• 2019

Rudbeckia laciniata var. hortensis Bailey is used in home remedy for colic and gastritis in South Korea. Although Rudbeckia laciniata var. hortensis Bailey is used extensively for home remedies, no single study on its efficacy exists. In this study, we investigated the anti-obesity effects of Rudbeckia laciniata var. hortensis Bailey. The anti-obesity effect of a 70% ethanol extract from Rudbeckia laciniata var. hortensis Bailey on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated with an Oil Red O assay, western blot analysis, and mRNA analysis. Compared to the control (only treated with DM), the 70% ethanol extract of Rudbeckia laciniata var. hortensis Bailey significantly inhibited adipocyte differentiation and intracellular triglyceride (TG) levels at a concentration of $100{\mu}g/ml$. To determine how the TG content was reduced, we measured the level of protein and mRNA expression of obesityrelated agents, such as peroxisome proliferators-activated receptor ${\gamma}$ ($PPAR{\gamma}$), CCAAT/enhancer- binding protein ${\alpha}$ ($C/EBP{\alpha}$), AMP-activated protein kinase (AMPK) phosphorylation, LPL, and FAS. As a result, the 70% ethanol extract of Rudbeckia laciniata var. hortensis Bailey significantly increased the expression of AMPK and decreased the expression of genes related to adipogenesis and fat storage, such as $PPAR{\gamma}$, $C/EBP{\alpha}$, LPL, and FAS.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.3
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• pp.316-322
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• 2019

Helicobacter pylori, a gram-negative bacterium, is one of the risk factors that induces gastritis and gastric cancer. Therefore, much attention has been paid to the compounds that inhibit bacterial growth or eradicate bacteria. Evodiae fructus (EF), the fruit of Evodia rutaecarpa, has been used for treating diarrhea and abdominal pain. EF extract was already found to inhibit the growth of H. pylori. However, to the best of our knowledge, the effect of EF on the virulence factors of H. pylori has not been reported. In this study, when comparing the minimum inhibitory concentration (MIC) of the different methanol concentration extracts, the 95% methanol extract (EF95) showed the lowest MIC value. EF95 extract suppressed the expressions of cagA, vacA and ureB, but interestingly, it up-regulated the expression of ureA. A decrease in production of ammonia in the culture medium and the cell lysates indicated that EF95 inhibited the urease activity in H. pylori, which was the result of EF95 inhibiting the ureB expression. Although the mechanism by which EF95 extract regulates the virulence factors in H. pylori needs further study, EF95 could be used for treatment of gastric troubles induced by H. pylori.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.255-264
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• 2018

This experiment was conducted to analyse the effects of flavone supplementation on the preimplantation development of in-vitro produced porcine embryos. During in-vitro development, immature oocytes and early embryos were exposed to different concentrations of flavone (0, $1{\mu}M$, $25{\mu}M$, $50{\mu}M$, and $100{\mu}M$ respectively). Results showed that $100{\mu}M$ of flavone significantly reduced the intracellular ROS levels of oocytes accompanied with a significant rise in GSH level. In parthenogenesis, no significant change was observed in the cleavage rates whether flavone was supplemented in IVM or IVC media. In IVM supplemented group, the blastocyst development rate was significantly enhanced by $1{\mu}M$ concentration than other groups (51.5% vs. 41.3%, 44.0%, 36.3%, 31.7%; P<0.05) respectively. However, in IVC group $1{\mu}M$ concentration significantly improved the blastocysts production than $50{\mu}M$ and control groups (50.0% vs. 40.5%, 38.0%; P<0.05) respectively. Following nuclear transfer, the cleavage rate of IVM group was significantly more in $1{\mu}M$ than $50{\mu}M$ and $100{\mu}M$ groups (92.9% vs. 89.7%, 87.8%; P<0.05), followed by similar pattern of cloned blastocysts production being significantly higher in $1{\mu}M$ group than $50{\mu}M$, $100{\mu}M$ and control groups (16.8% vs. 9.0%, 7.1%, 12.8%; P<0.05) respectively. In IVC group, $1{\mu}M$ concentration resulted in significantly higher cleavage rate than $25{\mu}M$ and $50{\mu}M$ groups (91.7% vs. 87.8%, 88.8%; P<0.05) respectively. However, the blastocysts production was significantly higher in $100{\mu}M$ group than others (26.2% vs. 13.6%, 14.0%, 18.2%; P<0.05) respectively. The optimal concentrations of flavone significantly enhanced the percentages of ICM:TE than control group (43.8% vs. 37.6%; P<0.05) accompanied with significantly higher expression levels of reprogramming related genes. In conclusion, the optimal concentrations of $1{\mu}M$ during IVM and $100{\mu}M$ during IVC can significantly improve the production of porcine in-vitro embryos.

• Journal of Life Science
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• v.29 no.1
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• pp.97-104
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• 2019

Streptococcus iniae is a typical fish pathogen causing streptococcosis and it can also cause zoonotic infectious diseases. We studied S. iniae FP5228 isolated from infected olive flounder in Wando, Korea. In a study to find virulence factors in FP5228, we found that the number of live bacteria decreased dramatically in culture medium containing S. iniae FP5228 for more than 24 hr. This phenomenon was hypothesized to be related to Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ genes, components of the Toxin/ Antitoxin (TA) system on the 14 kb plasmid of FP5228. We used a protein overexpression system to identify it. The pBP1140 vector system was constructed to regulate the expression of Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ by IPTG and Arabinose. E. coli/pBP1140 strain grew slowly in early growth under toxin expression condition, and it was confirmed by microscopic observation that the strain became longer. S. iniae CK287, lacking a 14 kb plasmid of S. iniae FP5228 strain, was constructed. CK287 bacterial cells did not show rapid killing during culture, and the ability to produce biofilm was also decreased, and toxicity was weakened in cytotoxicity test and fish test. These results suggest that the TA system is involved in physiological regulation and expression of virulence factors in S. iniae FP5228.

• Tuberculosis and Respiratory Diseases
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• v.82 no.2
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• pp.133-142
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• 2019

Background: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. Methods: Mouse AE II cell line MLE-12 were exposed to $1-10{\mu}g/mL$ BLM and $0.01-100{\mu}M$ baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. Results: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor ${\alpha}$, and transforming growth factor ${\beta}1$. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. Conclusion: BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.