• Title/Summary/Keyword: In vivo development

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Bioavailability and Anti-inflammatory Effect of Fermented Red Ginseng in BALB/c Mouse (BALB/c 마우스에서 발효 홍삼 Ginsenoside의 생체이용율과 항염효과)

  • Lee, Eun Kyu;Bae, Chu Hyun;Kim, Yu Jin;Park, Soo-Dong;Shim, Jae-Jung;Yu, Youngbob;Lee, Jung-Lyoul
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.433-442
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    • 2021
  • The fermented red ginseng by microorganism is known to increase pharmacological activity in vivo. To evaluate the bioavailablity of red ginseng fermented by probiotics, we conducted the pharmacokinetic study of ginsenoside Rb1, Rd and total ginsenosides (TG, ginsenosides Rb1 + Rd + Rg1 + F2 + Rg3 + compound K) in BALB/C mice. The AUC value of ginsenoside Rb1 in mice serum administered with 600mg/kg drugs showed 21.93 ± 14.68 ng·h/mL (RGw, water extract), 275.211 ± 110.04 ng·h/mL (RGe, 50% ethanol extract) and 404.91 ± 162.57 ng·h/mL (fRGe, fermented red ginseng extract). Analysis of ginsenoside Rd also showed a higher ACU value in fRGe than in RGw or RGe. And the AUC value of total ginsenosides in mice serum treated with 600 mg/kg were observed 42.12 ± 23.44 ng·h/mL (RGw), 321.44 ± 133.5 ng·h/mL (RGe) and 537.33 ± 229.01 ng·h/mL (fRGe), respectively. Cmax value of ginsenoside Rb1 in mice administered with 600mg/kg were observed 3.67 ± 3.34 ng/mL (RGw), 23.27 ± 8.81 ng/mL (RGe) and 25.52 ± 7.29 ng/mL (fRGe). These results can be considered that the fermented red ginseng has more bioavailability than that of unfermented red ginseng. In quantitative analysis of the inflammation-related cytokines IL-1β and TNF, no significant difference was found between the fermented red ginseng (fRGe) and the red ginseng (RGe).

Effects of Herbal Medicine Complex on Skin Inflammation and Atopic Dermatitis (한방 복합물이 피부 염증 및 아토피 피부염에 미치는 영향)

  • Ji-Hee, Choi;In-Hwan, Joo;Jong-Min, Park;Dong-Hee, Kim
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.5
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    • pp.187-192
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    • 2022
  • The purpose of this study is to examine the effect of herbal medicine complex (HMC) containing Camellia sinensis L., Duchoesna chrysantha, Houttuynia cordata Thunberg, Poncirus trifoliata Rafinesque on skin inflammation and atopic dermatitis. First, we examined the anti-inflammatory effect of HMC in TNF-α induced human keratinocytes (HaCaT cell). Real-time PCR and western blotting were performed to evaluate the expression of inflammatory cytokines (e.g., iNOS, COX-2, IL-6, IL-8) mRNA and protein. Four-weeks old male NC/Nga mice were treated with 1% 2,4-dinitrochlorobenzene (DNCB) solution and used as an atopic dermatitis mice model. And, HMC (200 mg/kg or 400 mg/kg) was administered directly into the stomach of mice for 4 weeks, and blood or serum analysis, tissue staining were performed after oral gavage. As a result HMC inhibited the mRNA expression of iNOS, COX-2, IL-6, and IL-8, which had been increased by TNF-α in HaCaT cells. In addition, the protein expression was also significantly suppressed in the same way as the mRNA expression results. The in vivo experiment results showed that, HMC administration reduced thickening of the epidermis and infiltration of eosinophil into the skin stratum basale compared to DNCB treatment. In addition, HMC administration significantly reduced the inflammatory cytokines (IL-4, IL-5, IL-6, and IL-13) production and immunocyte (white blood cell, lymphocyte, neutrophil, and eosinophil) count compared to DNCB treatment. Moreover, the serum IgE and histamine level was decreased by HMC administration. These results suggest that HMC can be used as effective herbal medicine extract for skin inflammation and atopic dermatitis. And this study may contribute to the development of the herbal medicine-based drug for the treatment of skin inflammation and atopic dermatitis.

Expression of Membrane-Type Matrix Metalloproteinase 1 and 2 in Mouse Oocytes, Embryos, Ovary and Oviduct (생쥐 난자와 배아 및 난소와 수란관의 Membrane-Type Matrix Metalloproteinase 1 및 2의 유전자 발현)

  • 김지영;이희진;김소라;김해권;강성구;이승재;조동제
    • Development and Reproduction
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    • v.4 no.1
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    • pp.45-52
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    • 2000
  • Membrane type matrix metalloproteinases(MT-MMPs) have been suggested to play an important role during structural remodeling of various tissue. Expression patterns of MT1-MMP and MT2-MMP mRNAs were investigated in oocytes, embryos, ovary and oviduct of mouse during their differentiation or periovulatory period using RT-PCR technique. Both cDNA products of MT1- and MT2-MMP of immature oocytes were barely discernable with a minimum amount but the expressions were distinct in mature oocytes regardless that they were matured in vivo or in vitro. MT2-MMP was not expressed by 2-cell embryos but was expressed by 4-cell stage embryos. From the morula stage untill hatched blastocyst stage, embyos showed intesnse expression of MT2-MMP with a sudden increase at blastocyst stage. While mouse ovarian tissues showed both expression of MT1- and MT2-MMP, there was no stage-specific difference throughout the estrous cycle. Mouse oviducts also exhibit constant amount of both MT1- and MT2-MMP expressions throughout periovulatory period, i.e., before or after ovulation. These observations lead to suggest that the differential expressions of maternal MT1- and MT2-MMP during meiotic resumption of mouse oocytes and embryonic expression of MT2-MMP particularly at blastocyst stage might play a role in the differentiation of mouse oocytes and/or embryos. The precise function of MT1- and MT2-MMP with regards to their participation in the remodeling of ovarian and oviductal tissues remains in a question.

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Rumen fermentation and performance of Hanwoo steers fed total mixed ration with Korean rice wine residue

  • Jeong, Chang-Dae;Mamuad, Lovelia L.;Ko, Jong Youl;Sung, Ha Guyn;Park, Keun Kyu;Lee, Yoo Kyung;Lee, Sang-Suk
    • Journal of Animal Science and Technology
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    • v.58 no.1
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    • pp.4.1-4.7
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    • 2016
  • Background: This study was conducted to evaluate the effects of adding Korean rice wine residue (RWR) in total mixed ration (TMR) on in vitro ruminal fermentation and growth performance of growing Hanwoo steers. Methods: For in vitro fermentation, the experimental treatments were Control (Con: 0 % RWR + TMR), Treatment 1 (T1: 10 % RWR + TMR), and Treatment 2 (T2: 15 % RWR + TMR). The rumen fluid was collected from three Hanwoo steers and mixed with buffer solution, after which buffered rumen fluid was transferred into serum bottles containing 2 g dry matter (DM) of TMR added with or without RWR. The samples were then incubated for 0 h, 12 h, 24 h, or 48 h at $39^{\circ}C$ and 100 rpm. For the in vivo experiment, 27 Hanwoo steers (6 months old) with an average weight of $196{\pm}8.66kg$ were subjected to a 24-week feeding trial. The animals were randomly selected and equally distributed into three groups. After which the body weight, feed intake and blood characteristics of each group were investigated. Results: The pH of the treatments decreased significantly relative to the control during the 12 h of incubation. Total gas production and ammonia nitrogen ($NH_3-N$) was not affected by RWR addition. The total volatile fatty acid (VFA) was lower after 24 h of incubation but at other incubation times, the concentration was not affected by treatments. Feed cost was 8 % and 15 % lower in T1 and T2 compared to control. Blood alcohol was not detected and a significant increase in total weight gain and average daily gain were observed in Hanwoo steers fed with RWR. Conclusion: Overall, the results of this study suggest that TMR amended with 15 % RWR can be used as an alternative feed resource for ruminants to reduce feed cost.

CK2 phosphorylates AP-2α and increases its transcriptional activity

  • Ren, Kaiqun;Xiang, Shuanglin;He, Fangli;Zhang, Wenfeng;Ding, Xiaofeng;Wu, Yanyang;Yang, Liping;Zhou, Jianlin;Gao, Xiang;Zhang, Jian
    • BMB Reports
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    • v.44 no.7
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    • pp.490-495
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    • 2011
  • Transcription factor AP-$2{\alpha}$ involves in the process of mammalian embryonic development and tumorigenesis. Many studies have shown that AP-$2{\alpha}$ functions in association with other interacting proteins. In a two-hybrid screening, the regulatory subunit ${\beta}$ of protein casein kinase 2 ($CK2{\beta}$) was identified as an interacting protein of AP-$2{\alpha}$; we confirmed this interaction using in-vitro GST pull-down and in-vivo co-immunoprecipitation assays; in an endogenous co-immunoprecipitation experiment, we further found the catalytic subunit ${\alpha}$ of protein casein kinase 2 ($CK2{\alpha}$) also exists in the complex. Phosphorylation analysis revealed that AP-$2{\alpha}$ was phosphorylated by CK2 kinase majorly at the site of Ser429, and such phosphorylation could be blocked by CK2 specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) in a dose-dependent manner. Luciferase assays demonstrated that both $CK2{\alpha}$ and $CK2{\beta}$ enhanced the transcription activity of AP-$2{\alpha}$; moreover, $CK2{\beta}$ increased the stability of AP-$2{\alpha}$. Our data suggest a novel cellular function of CK-2 as a transcriptional co-activator of AP-$2{\alpha}$.

Inhibition of Wntless/GPR177 suppresses gastric tumorigenesis

  • Seo, Jaesung;Kee, Hyun Jung;Choi, Hye Ji;Lee, Jae Eun;Park, Soo-Yeon;Lee, Seung-Hyun;Jeong, Mi-Hyeon;Guk, Garam;Lee, SooYeon;Choi, Kyung-Chul;Choi, Yoon Young;Kim, Hyunki;Noh, Sung Hoon;Yoon, Ho-Geun;Cheong, Jae-Ho
    • BMB Reports
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    • v.51 no.5
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    • pp.255-260
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    • 2018
  • Wntless/GPR177 functions as WNT ligand carrier protein and activator of $WNT/{\beta}$-catenin signaling, however, its molecular role in gastric cancer (GC) has remained elusive. We investigated the role of GPR177 in gastric tumorigenesis and provided the therapeutic potential of a clinical development of anti-GPR177 monoclonal antibodies. GPR177 mRNA expression was assessed in GC transcriptome data sets (GSE15459, n = 184; GSE66229, n = 300); protein expression was assessed in independent patient tumor tissues (Yonsei TMA, n = 909). GPR177 expression were associated with unfavorable prognosis [log-rank test, GSE15459 (P = 0.00736), GSE66229 (P = 0.0142), and Yonsei TMA (P = 0.0334)] and identified as an independent risk predictor of clinical outcomes: GSE15459 [hazard ratio (HR) 1.731 (95% confidence interval; CI; 1.103-2.715), P = 0.017], GSE66229 [HR 1.54 (95% CI, 1.10-2.151), P = 0.011], and Yonsei TMA [HR 1.254 (95% CI, 1.049-1.500), P = 0.013]. Either antibody treatment or GPR177 knockdown suppressed proliferation of GC cells and sensitized cells to apoptosis. And also inhibition of GPR177 suppresses in vitro and in vivo tumorogenesis in GC cells and inhibits $WNT/{\beta}$-catenin signaling. Finally, targeting and inhibition of GPR177 with antibody suppressed tumorigenesis in PDX model. Together, these results suggest GPR177 as a novel candidate for prognostic marker as well as a promising target for treatment of GC patients.

The Effect of Epigallocatechin-3-Gallate on Intimal Hyperplasia after Vascular Grafting (혈관이식술 후 내막과다증식에 대한 Epigallocatechin-3-Gallate의 효과)

  • Park, Han-Ki;Song, Suk-Won;Lee, Mi-Hee;Park, Jong-Chul;Joo, Hyun-Chul;Chang, Byung-Chul;Park, Young-Hwan
    • Journal of Chest Surgery
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    • v.40 no.4 s.273
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    • pp.256-263
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    • 2007
  • Background: Intimal hyperpiasia is characterized by a proliferation of vascular smooth muscle cells in the intimal layer Epigallocatechin-3-gallate (EGCG) is known to suppress smooth muscle cell proliferation. We propose that EGCG may have a protective effect against the development of intimal hyperplasia through the suppression of smooth muscle cell proliferation. Material and Method: Human umbilical vein endothelial cells (HUVEC) and rat aortic smooth muscle cells (RASMC) were cultured with different concentrations of EGCG, and proliferation and migration speed were measured. In 20 dogs, the autologous jugular veins were interposed into the carotid arteries. For the study group (n=10), the graft was stored for 30 minutes in EGCG solution and 300mM EGCG was applied to the perivascular space after grafting. After 6 weeks, the intimal and medial thickness was measured. Result: The proliferation of RASMC and HUVEC was suppressed with EGCG. The migration of RASMC was suppressed with EGCG, but that of HUVEC was not affected. In the in vivo study, the intimal thickness was thinner in EGCG group than in the control group (p<0.05), but the medial thickness did not show any difference. The intimal/medial thickness ratio was lower in the EGCG group (p<0.05). Conclusion: EGCG suppresses intimal hyperplasia after vascular grafting, and this may be mediated by prevention of migration and proliferation of vascular smooth muscle cells. The use of EGCG may offer new therapeutic modality to prevent intimal hyperplasia.

Effect of Radiotherapy on Chromosomal Aberration in Cancer Patients (암환자에서 방사선치료에의한 염색체이상)

  • Chun, Ha-Chung;Lee, Myung-Za;Yoo, Myung-Soo
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.43-50
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    • 1993
  • We evaluated frequency and types of chromosomal aberrations by ionizing radiation in cancer patients treated with radiotherapy in our institution. Twenty-five patients with various types of carcinomas such as lung, uterine cervix, esophagus, rectum, head and neck and pancreatic cancers were studied immediately before and after external beam radiotherapy. The frequency of aberrant metaphase prior to treatment was $4.93{\%}$, which was higher than that of control group. Especially in lung cancer, the freuqency of aberrant metaphase was three times higher than control group. A comparison of chromosomal abnormalities observed before and after radiotherapy demonstrated that proportion of aberrant rnetaphases was significantly inreased to $22.13{\%}$. Major chromosomal aberrations like structural abnormalities showed remarkalbe increase from 65.45 to $88.45{\%}$ after the treatment. Also the numbers of chromosomal alterations per cell were increased by a factor of 6.5. Aberrations with two or more break points were more prominently increased, compared with aberrations with single break point. The number of chromosomal break points was noted to be higher than expected value in No.1, 3, 8 and 11 chromosomes and lower in No.13, 15, 17 and 21 chromosomes. Based on this study, we believe that the distribution of chromosomal breakage is related with gene and chromosomal rearrangement which could result in the development of cancers.

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Development of Optical Molecular Imaging System for the Acquisition of Bioluminescence Signals from Small Animals (소동물 발광영상 측정을 위한 광학분자영상기기의 개발)

  • Lee, Byeong-Il;Kim, Hyeon-Sik;Jeong, Hye-Jin;Lee, Hyung-Jae;Moon, Seung-Min;Kwon, Seung-Young;Choi, Eun-Seo;Jeong, Shin-Young;Bom, Hee-Seung;Min, Jung-Joon
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.344-351
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    • 2009
  • Purpose: Optical imaging is providing great advance and improvement in genetic and molecular imaging of animals and humans. Optical imaging system consists of optical imaging devices, which carry out major function for monitoring, tracing, and imaging in most of molecular in-vivo researches. In bio-luminescent imaging, small animals containing luciferase gene locally irradiate light, and emitted photons transmitted through skin of the small animals are imaged by using a high sensitive charged coupled device (CCD) camera. In this paper, we introduced optical imaging system for the image acquisition of bio-luminescent signals emitted from small animals. Materials and Methods: In the system, Nikon lens and four LED light sources were mounted at the inside of a dark box. A cooled CCD camera equipped with a control module was used. Results: We tested the performance of the optical imaging system using effendorf tube and light emitting bacteria which injected intravenously into CT26 tumor bearing nude mouse. The performance of implemented optical imaging system for bio-luminescence imaging was demonstrated and the feasibility of the system in small animal imaging application was proved. Conclusion: We anticipate this system could be a useful tool for the molecular imaging of small animals adaptable for various experimental conditions in future.

Pharmacological Profile of KR-31125, an Orally Active AT1 Receptor Antagonist (안지오텐신 수용체 리간드 KR-31125의 생체 내 활성에 관한 연구)

  • Lee, Sung-Hou
    • Journal of Life Science
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    • v.20 no.7
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    • pp.969-976
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    • 2010
  • In vivo studies of KR-31125 (2-butyl-5-dimethoxymethyl-6-phenyl-7-methyl-3-[[2'-(1H-tetrazol-5-yl) biphenyl-4-yl]methyl]-3H-imidazo[4,5-b]pyridine) were performed in pithed rats, conscious angiotensin II (AII) challenged normotensive rats, renal hypertensive rats (RHRs) and furosemide-treated beagle dogs. KR-31125 induced a non-parallel right shift in the dose-pressor response curve to AII ($ID_{50}$: 0.095 mg/kg) with a dose-dependent reduction in the maximum responses in pithed rats. Compared to losartan, this antagonistic effect was about 18 times more potent, presenting competitive antagonism. Other agonists such as norepinephrine and vasopressin did not alter the responses induced by KR-31125. Orally administered KR-31125 had no agonistic effect and dose-dependently inhibited the pressor response to AII with a slightly weaker potency ($ID_{50}$: 0.25 and 0.47 mg/kg, respectively) in the AII-challenged normotensive rat model, but with a more rapid onset of action than losartan (time to $E_{max}$: 30 min for KR-31125 and 6 hr for losartan). KR-31125 produced a dose-dependent antihypertensive effect with a higher potency than losartan in RHRs, and these effects were confirmed in furosemide-treated dogs where they presented a dose-dependent and long-lasting (>8 hr) antihypertensive effect with a rapid onset of action (time to $E_{max}$: 2-4 hr), as well as a 20-fold greater potency than losartan. These results suggest that KR-31125 is a potent, orally active $AT_1$ receptor antagonist that can be applied to the development of new diagnostic and research tools as an added exploratory potential of $AT_1$ receptor antagonist.