• Journal of Embryo Transfer
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• v.29 no.4
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• pp.351-359
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• 2014

Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.

• Macromolecular Research
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• v.16 no.6
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• pp.510-516
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• 2008

New antitumor active polymers, poly(methacryloyl-2-oxy-1,2,3-propanetricarboxylic acid-co-exo-3,6-epoxy-l,2,3,6-tetrahydrophthalic acid) [poly(MTCA-co-ETAc)], poly(methacryloyl-2-oxy-l,2,3-propanetricarboxylic acid-co-hydrogen ethyl-exo-3,6-epoxy-l,2,3,6-tetrahydrophthalate) [poly(MTCA-co-HEET)], and poly(methacryloyl-2-oxy-l,2,3-propanetricarboxylic acid-co-a-ethoxy-exo-3,6-epoxy-1,2,3,6-tetrahydrophthaloyl-5-fluorouracil) [poly(MTCA-co-EETFU)] were synthesized and characterized. Their antitumor activity, inhibition of DNA replication and antiangiogenesis were examined. The structures of the polymers were identified by FT-IR, $^1H$ and $^{13}C$-NMR spectroscopy. The number average molecular weights of the fractionated polymers determined by GPC ranged from 9,400 to 14,900, and polydispersity indices were less than 1.7. The in vitro cytotoxicity of these polymers was determined and their antitumor activity was evaluated. The $IC_{50}$ values (the drug concentration at inhibition of 50% tumor growth) indicated that the synthesized polymers were much better inhibitors of cancer cells and showed lower cytotoxicity than the free 5-FU. The in vivo antitumor activity of the conjugates was examined using mice bearing the sarcoma 180 tumor cell line. The life spans (TIC) of the mice treated with the conjugates were higher than those treated with the free 5-FU. In addition, the synthesized conjugates showed excellent antiangiogenic activity based on an embryo chorioallantoic membrane assay.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.2
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• pp.12-22
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• 2012

Objectives: The aim of this study is to investigate immune enhancing effect of Houttuyniae Herba water extract(HW) on RAW 264.7 cell of mouse macrophages. Methods: Effects of HW on productions of nitric oxide(NO) and hydrogen peroxide($H_2O_2$) in RAW 264.7 mouse macrophages were measured. Effect of HW on production of cytokines such as interleukin(IL)-$1{\beta}$, IL-6, and tumor necrosis factor(TNF)-${\alpha}$ in RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. All of results were represented P<0.05 compared to the normal. Results: 1. After 24 hr incubation, HW increased significantly NO production in RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ${\mu}g$/mL. 2. After 24 hr incubation, HW increased significantly hydrogen peroxide production in RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ${\mu}g$/mL. 3. After 24 hr incubation, HW increased significantly IL-$1{\beta}$ production in RAW 264.7 cells at the concentrations of 100 and 200 ${\mu}g$/mL. 4. After 24 hr incubation, HW increased significantly IL-6 production in RAW 264.7 cells at the concentrations of 100 and 200 ${\mu}g$/mL. 5. After 24 hr incubation, HW increased significantly TNF-${\alpha}$ production in RAW 264.7 cells at the concentrations of 50, 100, and 200 ${\mu}g$/mL. Conclusions: These results suggest that HW has immune enhancing activity related with its increasement of NO, hydrogen peroxide, IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ in macrophages.

• Journal of fish pathology
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• v.17 no.1
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• pp.1-10
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• 2004

In order to analyse the detection of viral hemorrhagic septicemia virus (VHSV) in marine environment surrounding coastal region of Eastern and Southern sea of Korea, the pools of each organ sample of three fish were taken for virus assay from February to May in 2003. The samples comprised 42, taken from 9 species of marine fishes. The VHSV was detected from chub mackerel Scomber japonicus and striped mullet Mugil cephalus in epithelioma papulosum cyprini (EPC) cells. The identity of the virus was confirmed a reverse transcriptase-polymerase chain reaction (RT-PCR). VHSV has previously been reported from chub mackerel, but not from striped mullet. The new isolates was classified as a member of genogroup I (American type) of VHSV and was closely related to the VHSV KVHS'01-l based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene.

• Journal of Radiation Industry
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• v.5 no.1
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• pp.15-20
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• 2011

Protein kinases are the most important drug targets for the treatment of numerous diseases. The involvement of protein kinase C (PKC) in many biological processes such as development, memory, cell differentiation, and proliferation has been demonstrated. PKC is recognized as an important player in carcinogenesis. Thus, a variety of PKC inhibitors have been investigated. Among them, flavonoids have been demonstrated to affect the activity of many mammalian in vitro enzyme systems. The recent investigation was performed to evaluate the inhibitory effects of hesperidin, which is a flavonoid, on the proliferation and carcinogenesis of many cancers. In this study, an efficient kinase assay based on a biochip using radio-phosphorylation was established and performed for an examination of the inhibitory effects of hesperidin on PKC activity at different concentrations of 50, 200, 500 nM. It was found that hesperidin shows inhibitory potency on PKC, and that the biochip can be used to rapidly screen kinase inhibitors resulting in the therapeutic agents.

• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Journal of Dental Anesthesia and Pain Medicine
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• v.22 no.5
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• pp.369-376
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• 2022

Background: Nonobstetric surgery is sometimes required during pregnancy, and neck abscess or facial bone fracture surgery cannot be postponed in pregnant women. However, dental surgery can be stressful and can cause inflammation, and the inflammatory response is a well-known major cause of preterm labor. Propofol is an intravenous anesthetic commonly used for general anesthesia and sedation. Studies investigating the effect of propofol on human amnion are rare. The current study investigated the effects of propofol on lipopolysaccharide (LPS)-induced inflammatory responses in human amnion-derived WISH cells. Methods: WISH cells were exposed to LPS for 24 h and co-treated with various concentrations of propofol (0.01-1 ㎍/ml). Cell viability was measured using the MTT assay. Nitric oxide (NO) production was analyzed using a microassay based on the Griess reaction. The protein expression of cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE 2), p38, and phospho-p38 was analyzed using western blotting. Results: Propofol did not affect the viability and NO production of WISH cells. Co-treatment with LPS and propofol reduced COX-2 and PGE₂ protein expression and inhibited p38 phosphorylation in WISH cells. Conclusion: Propofol does not affect the viability of WISH cells and inhibits LPS-induced expression of inflammatory factors. The inhibitory effect of propofol on inflammatory factor expression is likely mediated by the inhibition of p38 activation.

• The Korea Journal of Herbology
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• v.28 no.5
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• pp.113-119
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• 2013

Objectives : The purpose of this study was to investigate the effects of Angelicae Gigantis Radix Water Extract(AG) on the production of proinflammatory mediators in RAW 264.7 cells stimulated with lipopolysaccharide(LPS). Method : RAW 264.7 cells were cotreated with AG(50 and 100 ug/mL) and lipopolysaccharide(LPS; 1 ug/mL) for 24 hours. After 24 hour treatment, using Bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, IL-$1{\beta}$, IL-10, tumor necrosis factor-alpha(TNF-${\alpha}$), granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), interferon inducible protein-10(IP-10), leukemia inhibitory factor(LIF), lipopolysaccharide-induced chemokine(LIX), monocyte chemoattractant protein-1(MCP-1), macrophage colony-stimulating factor(M-CSF), macrophage inflammatory protein(MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-2, Regulated on Activation, Normal T cell Expressed and Secreted(RANTES) and vascular endothelial growth factor(VEGF) were measured. Result : AG significantly inhibited LPS-induced production of TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, and M-CSF from LPS-stimulated RAW 264.7 cells at the concentrations of 50 and 100 ug/mL. AG significantly inhibited LPS-induced production of MIP-$1{\beta}$, MIP-2, GM-CSF, and IL-6 from LPS-stimulated RAW 264.7 cells at the concentrations of 50 ug/mL. AG significantly inhibited LPS-induced production of VEGF from LPS-stimulated RAW 264.7 cells at the concentrations of 100 ug/mL. But AG did not show any significant effect on the production of MCP-1, LIF, LIX, IP-10 and IL-$1{\beta}$ from LPS-induced RAW 264.7 cells. Conclusion : These results suggest that AG has anti-inflammatory effect related with its inhibition of proinflammatory mediators such as TNF-${\alpha}$, MIP-$1{\alpha}$, G-CSF, RANTES, IL-10, MIP-$1{\beta}$, MIP-2, GM-CSF, IL-6, VEGF and M-CSF in LPS-induced macrophages.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.40-40
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• 2023

Atopic dermatitis is a chronic inflammatory skin diseases caused by skin barrier dysfunction. Allium victoralis var. Platyphyllum (AVP) is a perennial plant used as vegetable and herbal medicine. The purpose of this study was to suggest that AVP is a new cosmetic material by examining the effects of AVP on the skin barrier and inflammatory response. A bibliometric network analysis was performed through keyword co-occurrence analysis by extracting author keyword from 69 articles retrieved from SCOPUS. We noted the anti-inflammatory activity shown by the results of clustering and mapping from network visualization analysis using VOSviewer software tool. HPLC-UV analysis showed that AVP contains 0.12 ± 0.02 mg/g of chlorogenic acid and 0.10 ± 0.01 mg/g of gallic acid. AVP at 100 ㎍/mL was shown to increase the mRNA levels of filaggrin and involucrin related to skin barrier function by 1.50-fold and 1.43-fold, respectively. In the scratch assay, AVP at concentrations of 100 ㎍/mL and 200 ㎍/mL significantly increased the cell migration rate and narrowed the scratch area. In addition, AVP suppressed the increase of inflammation-related factors COX-2 and NO and decreased the release of β-hexosaminidase. This study suggests that AVP can be developed as a functional cosmetic material for atopy management through skin barrier protection effects, anti-inflammatory and anti-itch effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.63-71
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• 2004

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98％ at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97％ at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79％ at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30％. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.