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Effects of Cryptotympana pustulata on the expression of cytokine genes in human monocytes of THP-1 (선퇴가 인간의 THP-1 단핵구에서 사이토카인 유전자 발현에 미치는 영향)

  • An, Jong-Hyun;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.94-110
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    • 2010
  • Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as phorbol-myristate-acetate(PMA), lipopolysaccharide(LPS), dermato-phagoides pteronyssus crude extract(DPE), dinitrochloro-benzene(DNCB) and Cryptotympana pustulata(CP), the Cryptotympana pustulata extracting substance at simultaneously on the translocation of nuclear factor-kappa B(NF-${\kappa}B$) towards to the nucleus and the mRNA expression patterns of various cytokine genes in Human acute monocytic leukemia cell line(THP-1 cells), monocytes of human. Experiment : To analyze cytokine genes expression patterns, the RT-PCR method was used, measuring tumor necrosis factor(TNF)-$\alpha$ that had been secreted during cell culture in the ELISA method. The morphological change in the cell observed during THP-1 cell culture was observed using a scanning electron microscope (SEM) and the quantitative distribution in the cell NF-${\kappa}B$ was analyzed through immunocytochemistry and a confocal microscopy. Result : CP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. The expressions of inter-leukin(IL)-10, interferon(INF)-$\gamma$, TNF-$\alpha$ and monocyte chemoattractantant protein(MCP)-1 induced by PMA were suppressed by CP while the expression of transforming growth factor(TGF)-$\beta$ was promoted. Regarding the secretion pattern of TNF-$\alpha$ according to PMA processing, its secretion amount was increased by CP concurrent processing, in case of processing CP onto PMA and LPS, We discovered that the secretion amount of TNF-$\alpha$ was increased. Upon processing PMA and LPS on the THP-1 cell strain at the same time or either additionally processing CP thereon, the movement increase towards the nucleus from the NF-${\kappa}B$ cell cytoplasm, a transcription factor was able to be observed. Conclusion : In this study, Cryptotympana pustulata extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances processed on the monocyte THP-1 cell of humans. Therefore, additional studies as for the immune adjusting function of Cryptotympana pustulata are considered to be able to offer important materials for curing immune abnormal diseases such as atopy dermatitis afterward.

Effects of Calcium and Indole-3-butyric acid Treatments on Calcium Concentration and Stem-End Browning in 'Fuyu' Sweet Persimmons (칼슘제 및 IBA 처리가 '부유' 단감과실의 칼슘함량 및 과정부 갈변현상에 미치는 영향)

  • Kim, Young;Kim, Wol-Soo;Choi, Hyun-Sug;Gu, Mengmeng
    • Food Science and Preservation
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    • v.16 no.4
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    • pp.459-462
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    • 2009
  • 'Fuyu' (Diospyros kaki L.) is an important sweet persimmon cultivar, and the fruits are often stored in a modified atmosphere after harvesting in South Korea. However, blossom-end browning and darkening of fruit often occur after harvest or during storage, which decreases fruit quality in the fresh fruit market. High fruit calcium concentration would reduce oxidation of phenolic compounds in the cytoplasm such oxidation is responsible for fruit browning. This study investigated the effects of soluble calcium fertilization and foliar application, and indole-3-butyric acid (IBA) fertilization on fruit quality and browning. Trees received one of the following five treatments: 1) control (no calcium or IBA); 2) calcium fertilization (Ca FG, 2 mL per tree); 3) calcium foliar application (Ca FA, 2 mL); 4) calcium and IBA fertilization (Ca+IBA) 5) IBA fertilization (IBA, 2 mL. Fruit calcium concentration was highest in trees treated by Ca FA, and lowest in control trees. Generally, fruit calcium concentration was high in the stem end but low in the blossom end, which usually first develops fruit-browning symptoms. There were no apparent differences in fruit qualities such as firmness, soluble solid content, and weight among treatments. Fruit browning occurred at frequencies of about 14%, 20%, and 50% on Ca FA, Ca FG, and control trees, respectively. Therefore, the improved fruit calcium level seen when trees received Ca or IBA application tended to prevent fruit browning, which increased fruit quality and storage properties.

Chicken FMRP Translational Regulator 1 (FMR1) Promotes Early Avian Influenza Virus Transcription without Affecting Viral Progeny Production in DF1 Cells

  • Woo, Seung Je;Park, Young Hyun;Han, Jae Yong
    • Korean Journal of Poultry Science
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    • v.48 no.2
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    • pp.81-90
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    • 2021
  • Avian influenza viruses (AIVs) must utilize host cellular factors to complete their life cycle, and fragile X mental retardation protein (FMRP) has been reported to be a host factor promoting AIV ribonucleoprotein (vRNP) assembly and exports vRNP from the nucleus to the cytoplasm. The functional role of chicken FMRP translational regulator 1 (cFMR1) as a host factor of AIV is, however, poorly understood. In this study, we targeted the cFMR1 gene in DF1 cells using clustered regularly interspaced short palindromic repeats/Cas9-mediated genome editing to examine the functional role of cFMR1 as a host factor of AIV. We found that cFMR1 stimulated viral gene transcription during early stages of the viruses' life cycle and did not affect viral progeny production and viral polymerase activity in DF1 cells 24 hours post infection. cFMR1 overexpression did not exert significant effects on virus production, compared to the control. Therefore, unlike in mammalian systems (e.g., humans or mice), cFMR1 did not play a pivotal role in AIV but only seemed to stimulate viral proliferation during early stages of the viral life cycle. These results imply that the interplay between host factors and AIV differs between mammals and avian species, and such differences should be considered when developing anti-viral drugs for birds or establishing AIV-resistant bird models.

The Effect of Nicotine on the Proliferation and Differentiation of Normal Human Osteoblast at the Surface of Implants (임플란트 표면에서 배양된 정상인 조골세포의 증식 및 분화에 미치는 니코틴의 영향)

  • Ahn, Tae Woong;Lee, Chong Heon
    • The Korean Journal of Oral and Maxillofacial Pathology
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    • v.42 no.5
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    • pp.111-118
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    • 2018
  • Nicotine of tobacco component has a controversial impact in the clinical outcome of dental implants. Although numerous nicotine effects on bone healing around implants have been presented, it is rarely reported in vitro study about normal human osteoblast(NHost) from oral and maxillofacial area at the surface of implants. The purpose of the present study was to evaluate the effect of nicotine on the proliferation and differentiation response of NHost to plasmatic and salivary levels of nicotine reported in smokers at the surface of screw-type plasma-sprayed titanium implants. NHosts were seeded on the surface of titanium implants and cultured for 21 days in ${\alpha}-MEM$ supplemented with 10% FBS, 50mg/ml ascorbic acid, 5mM ${\beta}$-glycerophosphate and 100nM dexamethasone. Seeded implants were exposed to various nicotine concentration(0.05-0.5mg/ml) from 1 to 21 days, and characterized for cell morphology, proliferation, differentiation, alkaline phosphatase(ALP) activity and ionized calcium concentration(Cai) of medium. Continuous exposure to higher nicotine concentration(above 0.3mg/ml) induced a dose- and time-dependent vacuolation of the cytoplasm, and a tendency to detach from the implant surface. 0.05mg/ml(lower nicotine concentration) did not cause significant effects in the cell proliferation and ALP activity. 0.1-0.2mg/ml caused evident dose-dependent effects in increased cell proliferation, ALP activity and earlier onset of matrix mineralization at levels up to 0.2mg/ml, while a dose-dependent inhibitory effect at 0.3-0.5mg/ml. Cai concentration of control group was decreased at 14 days. Increased Cai concentration at 0.1-0.2mg/ml, decreased Cai concentration at 0.3mg/ml and no change at 0.5mg/ml during the culture period were seen. It suggested that nicotine concentration could paly an role in modulating NHost activity as a contributing factor associated with proliferation and differentiation of NHost at the surface of implants.

The role of p21/CIP1/WAF1 (p21) in the negative regulation of the growth hormone/growth hormone receptor and epidermal growth factor/epidermal growth factor receptor pathways, in growth hormone transduction defect

  • Kostopoulou, Eirini;Gil, Andrea Paola Rojas;Spiliotis, Bessie E.
    • Annals of Pediatric Endocrinology and Metabolism
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    • v.23 no.4
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    • pp.204-209
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    • 2018
  • Purpose: Growth hormone transduction defect (GHTD) is characterized by severe short stature, impaired STAT3 (signal transducer and activator of transcription-3) phosphorylation and overexpression of the cytokine inducible SH2 containing protein (CIS) and p21/CIP1/WAF1. To investigate the role of p21/CIP1/WAF1 in the negative regulation of the growth hormone (GH)/GH receptor and Epidermal Growth Factor (EGF)/EGF Receptor pathways in GHTD. Methods: Fibroblast cultures were developed from gingival biopsies of 1 GHTD patient and 1 control. The protein expression and the cellular localization of p21/CIP1/WAF1 was studied by Western immunoblotting and immunofluorescence, respectively: at the basal state and after induction with $200-{\mu}g/L$ human GH (hGH) (GH200), either with or without siRNA CIS (siCIS); at the basal state and after inductions with $200-{\mu}g/L$ hGH (GH200), $1,000-{\mu}g/L$ hGH (GH1000) or 50-ng/mL EGF. Results: After GH200/siCIS, the protein expression and nuclear localization of p21 were reduced in the patient. After successful induction of GH signaling (control, GH200; patient, GH1000), the protein expression and nuclear localization of p21 were reduced. After induction with EGF, p21 translocated to the cytoplasm in the control, whereas in the GHTD patient it remained located in the nucleus. Conclusion: In the GHTD fibroblasts, when CIS is reduced, either after siCIS or after a higher dose of hGH (GH1000), p21's antiproliferative effect (nuclear localization) is also reduced and GH signaling is activated. There also appears to be a positive relationship between the 2 inhibitors of GH signaling, CIS and p21. Finally, in GHTD, p21 seems to participate in the regulation of both the GH and EGF/EGFR pathways, depending upon its cellular location.

Increased Apoptotic Efficacy of Decitabine in Combination with an NF-kappaB Inhibitor in Human Gastric Cancer AGS Cells (핵산합성 억제제인 decitabine과 NF-κB 활성 저해제인 PDTC의 병용 처리에 의한 인체 위암세포사멸 효과 증진)

  • Choe, Won Kyung;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1268-1276
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    • 2018
  • The cytidine analog decitabine (DEC) acts as a nucleic acid synthesis inhibitor, whereas ammonium pyrrolidine dithiocarbamate (PDTC) is an inhibitor of nuclear factor-${\kappa}B$. The aim of this study was to investigate the possible synergistic inhibitory effect of these two inhibitors on proliferation of human gastric cancer AGS cells. The inhibitory effect of PDTC on AGS cell proliferation was significantly increased by DEC in a concentration-dependent manner, and this inhibition was associated with cell cycle arrest at the G2/M phase and the induction of apoptosis. This induction of apoptosis by the co-treatment with PDTC and DEC was related to the induction of DNA damage, as assessed by H2AX phosphorylation. Further studies demonstrated that co-treatment with PDTC and DEC induced the disruption of mitochondrial membrane potential, increased the generation of intracellular reactive oxygen species (ROS) and the expression of pro-apoptotic Bax, and down-regulated the expression of anti-apoptotic Bcl-2, ultimately resulting in the release of cytochrome c from the mitochondria into the cytoplasm. Co-treatment with PDTC and DEC also activated caspase-8 and caspase-9, which are representative caspases of the extrinsic and intrinsic apoptosis pathways. Co-treatment also activated caspase-3, which was accompanied by proteolytic degradation of poly (ADP-ribose) polymerase. Taken together, these data clearly indicated that co-treatment with PDTC and DEC suppressed the proliferation of AGS cells by increasing DNA damage and activating the ROS-mediated extrinsic and intrinsic apoptosis pathways.

Characteristics of Viral Endothelial Cell Necrosis of Eel (VECNE) from Culturing Eel (Anguilla japonica, Anguilla bicolar) in Korea (국내 양식산 뱀장어(Anguilla japonica, Anguilla bicolar)에서 발생하는 바이러스성내피세포괴사증(VECNE)의 감염 특성)

  • Kim, Su Mi;Ko, Sang Mu;Jin, Ji Hye;Seo, Jung Soo;Lee, Nam Sil;Kim, Young Suk;Gu, Jeong Hui;Bae, Yu Ri
    • Korean Journal of Ichthyology
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    • v.30 no.4
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    • pp.185-193
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    • 2018
  • From 2017 to 2018, the disease has been monitored at four culturing eel farms in Incheon and Gyeonggi region in Korea. As a result, diseases with gill congestion frequently occurred. This disease occurred regardless of size of eel, but the frequency and cumulative mortality were high in eels within 3 months after stocking. The infected fish showed pathological histopathological features such as intense congestion and dilation in the central venous sinus (CVS) of gill filaments and hemorrhages in liver and kidney. Hexagonal viral particles measuring about 70 nm in diameter was observed in nuclei and cytoplasm of gill vascular endothelial cells. Molecular biologic diagnosis by both PCR and genetic analysis has been revealed that the causative agent of this disease is Japanese eel endothelial cells-infecting virus (JEECV), the cause of viral endothelial cell necrosis of eel (VECNE), which is mainly reported in Japan. This study is the first report on the characteristics of JEECV and VECNE infection in domestic eel farms.

Anti-inflammatory effect of Euonymi Lignum Suberalatum in LPS-activated Raw 264.7 cells and CA-induced paw edema rat model (귀전우(鬼箭羽)의 in vitro 및 in vivo에서의 염증억제효과)

  • Jeon, Chang Kwon;Park, Sang Mi;Park, Chung A;Byun, Sung Hui;Kim, Sang Chan
    • Herbal Formula Science
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    • v.27 no.2
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    • pp.101-120
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    • 2019
  • PURPOSE : Euonymi Lignum Suberalatum (EL) is the stem fin of Euonymi alatus. In traditional Korean medicine, EL is used for treatment of uterine bleeding, metritis and static blood. Recently, many studies have reported several pharmacological effects of EL including anticancer, antimicrobial, antidiabetic activity, and anti-oxidative stress. However, the mechanisms underlying anti-inflammatory effects by the EL is not established. METHODS : To investigate anti-inflammatory effects of Euonymi Lignum Suberalatum Water (ELWE), Raw 264.7 cells were pre-treated with $10-300{\mu}g/mL$ of ELWE, and then exposed to $1{\mu}g/mL$ of LPS. Levels of NO, IL-6, $IL-1{\beta}$ and $TNF-{\alpha}$ were detected by ELISA kit. Expression of pro-inflammatory proteins were determined by immunoblot analysis. To evaluate the anti-inflammatory effect in vivo, rat paw edema volume, and expressions of COX-2 and iNOS proteins in carrageenan (CA)-induced rat paw edema model. RESULTS : NO production activated by LPS, was decreased by $30-300{\mu}g/mL$ of ELWE. Production of inflammatory mediators such as $TNF-{\alpha}$, ILs, $PGE_2$ were decreased by ELWE 100 and $300{\mu}g/mL$. In addition, ELWE reduced LPS-mediated iNOS and COX-2 expression. Moreover, ELWE increased $I-{\kappa}B{\alpha}$ expression in cytoplasm and decreased $NF-{\kappa}B$ expression in nucleus. In vivo study, ELWE reduced the increases of paw swelling, and expression of iNOS and COX-2 proteins in paw edema induced by CA injection. CONCLUSION : The results indicate that ELWE could inhibit the acute inflammatory response, via modulation of $NF-{\kappa}B$ activation. Furthermore, inhibition of rat paw edema induced by CA is considered as clear evidence that ELWE may be a useful source to treat acute inflammation.

Anti-Inflammatory Effect of Ethanol Extract from the Seeds of Arctium Lappa L. in Vascular Endothelial Cells (혈관내피세포에서 우방자(牛蒡子) 에탄올 추출물의 항염증 효과)

  • Lee, Yun-Jung;Yoon, Jung-Joo;Kim, Hye-Yoom;Ahn, You-Mee;Hong, Mi-Hyeon;Son, Chan-Ok;Na, Se-Won;Lee, Ho-Sub;Kang, Dae-Gill
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.3
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    • pp.20-31
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    • 2019
  • Objectives: The seeds from Arctium lappa have been considered for its various pharmacological properties, which include anti-carcinogenic, anti-inflammatory, anti-diabetic, and anti-viral activities. Methods: In the present study, we investigated the anti-inflammatory effect of the ethanol extract from the seeds of Arctium lappa L (EAL) on cytokine-induced vascular inflammation in human umbilical vein endothelial cells (HUVEC). Results: Pretreatment with EAL significantly decreased tumor necrosis factor alpha ($TNF-{\alpha}$)-induced cell adhesion molecules expression such as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial-selectin (E-selectin) in a dose-dependent manner. Cell adhesion assay showed that pretreatment with EAL suppressed HUVEC-monocyte adhesion by $TNF-{\alpha}$ over $1{\mu}g/ml$ concentration. We investigated the involvement of nuclear transcription factor kappa-B ($NF-{\kappa}B$) in $TNF-{\alpha}$-induced vascular inflammation. $NF-{\kappa}B$ p65 nuclear expression was induced by $TNF-{\alpha}$, however, pretreatment with EAL was attenuated that nuclear translocation. In cytoplasm, EAL was also attenuated $TNF-{\alpha}$-induced decrease of inhibitor of ${\kappa}B-{\alpha}$ ($I{\kappa}B-{\alpha}$) expression. Moreover, EAL significantly decreased $TNF-{\alpha}$-induced production of intracellular reactive oxygen species (ROS). Conclusions: Taken together, our findings suggest that seeds of Arctium lappa L could be a therapeutic herb for prevention of cardiovascular diseases throughout the inhibition of vascular endothelial inflammation.

Changes in the Concentration and Localization of Accumulated Mercury in Kidney, Liver, and Spleen of Mice over Time (생쥐 신장, 간, 비장 내 시간에 따른 수은 농도 변화와 수은 화합물의 위치)

  • Kim, Yu Seon;Kim, Young Eun;Cho, Hyun Wook
    • Journal of Life Science
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    • v.29 no.8
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    • pp.879-887
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    • 2019
  • This study investigated the localization and changes in the concentration of injected mercury in the kidney, liver, and spleen of mice. To evaluate changes in the concentration of mercury over time, the mice were euthanized 10, 150, and 300 days post-treatment. Localization of accumulated mercury was identified by the autometallography method. Mercury was densely located in the supranuclear cytoplasm of epithelial cells of proximal tubules of the kidney but was not detected in the glomerulus 10 days post-treatment. In the liver, mercury was mainly found in hepatocytes around the portal vein and in sinusoidal Kupffer cells 10 days post-treatment. Mercury was scattered throughout both white and red pulp of the spleen 10 days post-treatment. In terms of changes in the concentration of mercury, the levels were lower in the renal cortex and medulla 150 and 300 days post-treatment as compared with those 10 days post-treatment. Mercury was found at low concentrations in liver hepatocytes 150 and 300 days post-treatment. The mercury concentration was also low in both the white and red pulp of the spleen 150 and 300 days post-treatment. Therefore, the concentrations of accumulated mercury in the kidney, liver, and spleen 150 and 300 days post-treatment were lower than those 10 days post-treatment. We identified the localization of mercury in cells and tissues of several organs and observed that accumulated mercury in organs decreased naturally over time.