• Title/Summary/Keyword: ICR cell

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Biological Activity of Recombinant Human Thrombopoietin (유전자 재조합 인간의 Thrombopoietin의 생리활성)

  • Kim, Boing-Soon;Naidansuren, Purevjargal;Min, Kwan-Sik
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1497-1504
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    • 2007
  • To investigate the function and secretion of human thrombopoietin (TPO) in mammalian cells, hTPO cDNA was cloned using human liver cDNA, and recombinant hTPO (rec-hTPO) was produced in CHO cell lines. In addition, six N-linked glycosylation sites were substituted for Ala to elucidate the role of each carbohydrate chain. To analyze the biological activity, rec-hTPO protein was injected subcutaneously. Blood was withdrawn for platelet determination. The metabolic clearance rate (MCR) was also analyzed at the 1, 4, 10 and 24 hr after tail vein injection. Wild-type TPO (WT) was efficiently secreted into the medium. However, a hTPO mutant with 116 deleted nucleotides detected by PCR cloning was not secreted. The N-linked glycosylation sites had nearly the same expression quantity as rec-hTPO WT apart from mutants 3 and 4. The glycosylation site of mutant 4 appeared to be an indispensable site for hTPO secretion. Also characterized was the biological activity through an injection with rec-hTPO (10 ng) to ICR mice (7 weeks). The result of the blood analysis showed a considerable increase in the platelet number six days after He injection. To analyze the pharmacokinetics, rec-hTPO was injected into the tail vein (5 ng). The result was 200 pg/ml 1hr after this injection. Following this, it dramatically decreased and virtually disappeared 10 hours after the injection. Thus, rec-hTPO may be a treatment for thrombopenia by the production of the high active rec-hTPO. In addition, hTPO can permit the development of potent new analogues that stimulate the platelet value.

Immunostimulating and Anti-cancer Effects of Pediococcus pentosaceus EROM101 Isolated from Korea. (한국인으로부터 분리한 Pediococcus pentosaceus EROM101의 면역증강 및 항암활성)

  • 송미경;우석규;장정순;김중학;김화영;홍성길;이병욱;박미현;정건섭
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.355-361
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    • 2003
  • Immunostimulating effects of lactic acid bacteria as biological response modifier is a subject of growing interest, but the knowledge of these focused on some bacteria as Lactobacillus and Bifidobacterium. In this study, we investigated the effects of Pediococcus pentosaceus EROM101 on the immunostimulating and anti-cancer activity in murine model. P. pentosaceus was mainly found in Kimchi and fermented sea food and is facultatively anaerobic, catalase-netative, gram-positive cocci arranged in pairs, tetrads and clusters. The immunostimulating effects of P. pentosaceus EROM101 were evaluated using IgA production assay of Peyer's patch and proliferation assay of exudated immune cells of Balb/C mice fed P. pentosaceus EROM101 for 3 weeks. The macrophage and splenocyte proliferation were enhanced by orally administrated of P. pentosaceus EROM101. Also, IgA production in Peyer's patch increased by P. pentosaceus EROM101. Anti-cancer activity of P. pentosaceus EROM101 was appeared in Sarcoma 180 tumor-bearing ICR mice. However, this bacterium lysate itself appeared to have noncytotoxic substance against Sarcoma 180 cell in vitro. These results suggested that P. pentosaceus EROM101 reinforce immune system and therefore was revealed to be anti-cancer activity in mice.

Anti-inflammatory Effects of Hongdeung-Tang on Mice with LPS Injected in Vagina (홍등탕(紅藤湯)이 LPS 질내(膣內) 주입(注入) 생쥐에 미치는 항염증(抗炎症)효과)

  • Seo, Ji-Young;Kim, Yoon-Sang;Yun, Jeong-Mun;Lee, Tae-Hui;Lim, Eun-Mee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.1
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    • pp.81-93
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    • 2005
  • Purpose : There are various oriental medicine therapy of treating Pelvic Inflammatory Disease(PID) in clinics. We made the PID mice model by injection Lipopolysaccharide (LPS) in vagina, and investigate anti-inflammatory effects of Hongdeung-Tang among oral medication, retention enema therapy and herbal-acupuncture treatment. Method : The ICR(20-30g) mice(♀) were used. To examine the occurrence of inflammation, LPS in different concentration was injected into the vagina of the mice, and White Blood Cell(WBC) in blood was counted. To examine anti-inflammatory effects, 6 mice were assigned to each of the normal group, the control group and the sample group. Hongdeung-Tang was medicated in oral and rectal with 1.0g/kg low dose and 3.0g/kg high dose and by herbal acupucture for 5 days, 2 days before LPS injection. After 3days from LPS injection, blood was collected from retro-orbital plexus, and WBC, Interleukin-6(IL-6), Tumor Necrosis Factor-${\alpha}$(TNF-${\alpha}$) in the blood was counted. Result : After LPS injection with each dose, WBC count showed significant increase depending on LPS concentration from $100{\mu}g/kg$, and it was maximized at 3 or 4 days after LPS injection. the Hongdeung-Tang treatment groups, The number of WBC was decreased significantly only in low dose and high dose oral medication, and IL-6 concentration showed significant decrease in oral and rectal medication as well as in herbal acupuncture treatment. TNF-${\alpha}$ concentration was decreased significantly in oral and rectal medication of low dose and high dose. herbal-acupuncture treatment group datas showed reductive tendency. Conclusion : Based on above results, we made the PID mice model by injection LPS in vagina, and demonstrated anti-inflammatory effect of Hongdeung-Tang of oral medication, retention enema therapy and herbal-acupuncture treatment.

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Studies on Antitumor Components of Collybia confluens (밀버섯의 항암성분에 관한 연구)

  • Kim, Sook-Hee;Kim, Jin-Sook;Jin, Mi-Rim;Kim, Ha-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • Korean Journal of Pharmacognosy
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    • v.24 no.4
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    • pp.267-281
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    • 1993
  • To find antitumor components from higher fungi, the mycelia of Collybia confluens (Pers. ex Fr.) Kummer were cultured in artificial media. For efficient production of the mycelia, the influences of various modifications of culture conditions were examined. A water-soluble protein-bound polysaccharide fraction, Fr. A, was obtained from the mycelia by hot water extraction. When Fr. A was purified and fractionated by DEAE-cellulose and Sepbadex G-200 gel filtration chromatographies into four fractions which were designated B, C, C-I and C-II. The tumor inhibition ratios of these fractions ranged from 46% to 75% against the solid forms of sarcoma 180 in ICR mice at doses of 20 and 50 mg/kg/day when given intraperitoneally. Especially, Fr. C which was named Collyban(CB) exhibited a marked life-prolonging effect of the mice against ascitic forms of sarcoma 180 at a dose of 50 mg via i.p. administration. To extend spectra of the antitumor activities and eliminate the effects of allograft rejection, the characterization of antitumor effects of CB was performed in syngeneic host-tumor systems. It did not show any antitumor activity against L1210 murine leukemia in $CD_2Fl$ mice but prolonged their life span against ascitic forms of $MM_{46}$ carcinoma in $C_3H/He$ mice. Also it exhibited antitumor activity against human cervical cancer HeLa cells that were xenografted into nude mice having BALB/c genetic backgrounds by the i.p. injection at a dose of 100 mg/kg/day. In order to characterize the antitumor components, CB was examined by chemical analysis. It was acidic protein-bound polysaccharides composed of 31% polysaccharide, 27% protein and 3% hexosamine. CB was fractionated into two fractions, Fr. C-I(M.W.: 500 Kd) and Fr. C-II(M.W.:30 and 8 Kd) by Sephadex G-200 gel filtration chromatography.

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Effect of Dendropanax morbifera extract Addition on Chungkukjang on Immune Response of Mice (황칠 추출물을 첨가한 청국장이 마우스의 면역기능에 미치는 효과)

  • Kim, Jin-Sol;Cheong, Min-Ju;Chung, Kyoung-A;Song, Seon-Young;Lee, Hyun-Hwa
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.6
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    • pp.430-438
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    • 2019
  • This study is to investigate increase immunomodulatory activity of Chungkukjang added with Dendropanax morbifera extract. There are four groups divided that Control, Dendropanax morbifera extract group, Chungkookjang group and Chungkukjang added to Dendropanax morbifera extract group which of five mouses each group. Their immunological characteristics were compared with either general Chungkukjang or non treated group in the tested male ICR mice. There were no significant difference in body weight and organ weight. In the plaque forming cell assay, Dendropanax morbifera extract 250 and 500 mg/kg groups measured significant increase over the control group. The values of lymphocytes were significantly increased in the Dendropanax morbifera extract 500 mg/kg group campared with the control group. In the general Chungkukjang 400 mg/kg added with Dendropanax morbifera extract 500 mg/kg group, total serum immunoglobulin G concentration was significantly higher than the control group and their spleen tissues observed proliferation of white puls. These results demonstrated that Chungkukjang Added with Dendropanax morbifera extract was provided enhance of immunomodulatory activity and suggest that it can be used as various functional foods, based on foods promote immune system health.

Anti-inflammatory and Anti-Pruritonic Effects of WSY-1075 composited with Medicinal Plants on the Activated Rat Peritoneal Mast cells and Mouse Pruritus (활성화된 렛트 비만세포와 마우스 소양증에 대한 한약재로 조성된 WSY-1075의 항염증 및 항소양 효과)

  • Hwang, Sung Yeoun;Lee, Seung Ho;Lee, Chia Wei;Kim, Jang Ho;Jang, Seon Il;Kim, An Na;Kim, Hong Jun
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.93-100
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    • 2013
  • Objectives : This study was to evaluate the anti-inflammatory and anti-pruritic effects of WSY-1075 composited with Corni Fructus, Angelica gigantis Radix, Lycii Fructus, Ginseng Radix, Cervi parvum Cornu and Cinnamomi Cortex in rat peritoneal mast cells (RPMCs) and scratching mouse model. Methods : WSY-1075 was prepared by extracting with 30% ethanol. In the present study, we investigated the effect of WSY-1075 on the production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and histamine in rat peritoneal mast cells (RPMCs) activated with phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187, and on the scratching behavior in mice treated with pruriogens. Results : WSY-1075 was not cytotoxic effect in used all concentration. PMA plus A23187 treatment significantly increased TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 production compared with media control in RPMCs. However, TNF-${\alpha}$, $IL1{\beta}$ and IL-6 production increased by PMA plus A23187 treatment were significantly inhibited by WSY-1075 (200 ${\mu}g/mL$ and 400 ${\mu}g/mL$). WSY-1075 also inhibited the histamine release from RPMCs stimulated by compound 48/80, which promotes histamine release. Moreover, WSY-1075 administration had an inhibitory effects on the scratching behavior induced by pruritogen (compound 48/80, histamine, serotonin and substence P) in ICR mice. Conclusion : These results suggest that WSY-1075 administration (200 mg/kg or 400 mg/kg) has the anti-inflammatory and anti-pruritic effects on the activated rat peritoneal mast cell and mouse pruritus. WSY-1075 has a potential use as a composition of medicinal plants for treatment against inflammation- and pruritus-related disease.

Hepatoprotective effect of fermented Schizandrae Fructus Pomace extract and Hoveniae Semen Cum Fructus extract combination mixtures against carbon tetrachloride-induced acute liver injured mice (사염화탄소 유발 급성 간 손상에 대한 발효 오미자박 및 헛개과병 추출물의 혼합 비율에 따른 간 보호효능)

  • Hye-Rim, Park;Kyung Hwan, Jegal;Beom-Rak, Choi;Jae Kwang, Kim;Sae Kwang, Ku
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.53-65
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    • 2023
  • Objectives : Present study investigated the hepatoprotective effects and the optimal mixing ratio of fermented Schizandrae Fructus Pomace (fSFP) and Hoveniae Semen Cum Fructus (HSCF) extract combination in carbon tetrachloride (CCl4)-induced acute liver injury mice. Methods : ICR mice were orally administered with 200 mg/kg of fSFP, HSCF and mixtures of fSFP and HSCF [MSH (w:w); 1:1, 1:2, 1:4, 1:6, 2:1, 4:1, 6:1, and 8:1] for 7 consecutive days. Silymarin (100 mg/kg) was administered as a reference drug. 0.5 mL/kg of CCl4 was injected intraperitoneally to induce acute liver injury. Body weight gain, relative liver weight, serum chemistry, histopathological analysis, and hepatic endogenous antioxidants capacities were observed. Results : All diverse combinations of MSH significantly reduced relative liver weight increase by CCl4. In addition, MSH administrations significantly decreased the elevation of serum alanine aminotransferase and aspartate aminotransferase activities by CCl4. Histopathological observation indicated that all MSH treatments significantly reduced the increase of degenerated hepatocytes, inflammatory cell infiltration, and histological activity index score by CCl4. Moreover, all MSH administrations reduced the elevation of malondialdehyde contents, and ameliorated the reduction of hepatic glutathione contents, superoxide dismutase activity, and catalase activity. Among the various mixing ratio of MSH combinations, MSH 1:1 and 2:1 showed the most potent anti-oxidative stress, and hepatoprotective effect. Conclusion : Present results suggest that 1:1 and 2:1 combinations of MSH is promising herbal formulation with the hepatoprotective effect against oxidative stress.

Dysregulation of Cellular Immune Functions on Gastric Administration to Mixtures of Polyethlene Microplastics and Metallic Lead in Mice (폴리에틸렌 미세플라스틱과 납의 복합노출에 따른 실험동물의 세포면역기능 조절 장애 평가)

  • Gyoungwoo Lee;Changyul Kim
    • Journal of Environmental Health Sciences
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    • v.49 no.1
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    • pp.22-29
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    • 2023
  • Background: The existing research results on the combined toxicity of these pollutants using mammals, such as rodents, are insufficient, especially in relation to changes in the immune system. Objectives: This study aims at evaluating the cellular immune response to PE-MPs solely or when combined with Pb, which possess excellent adsorption capacity with PE-MPs and is commonly co-exposed in our daily lives. Methods: The study investigated the cellular immune function of 9-week ICR mice with 28 days exposure to PE-MPs (2 mg/mouse/day) and Pb (0.1 mM in distilled water) individually and in combination. PE-MPs were administered via gastric intubation while the lead intake was conducted via the oral drinking water route. Cellular immunity was evaluated by analyzing the production for TH1 cytokines namely, TNF-α and IFN-𝛾 and TH2 cytokines, IL-4 and IL-6 in culture supernatants from polyclonally activated splenic mononuclear cells ex vivo. Results: Both the PE-MPs only and the PE-MPs+Pb exposure group revealed an increased TH1 response with elevated TNF-α and IFN-𝛾 levels and downregulated TH2 response with low IL-4, and IL-6 production levels compared to the control group. Furthermore, an increased IFN-𝛾/IL-4 ratio was found in the PE-MPs only and PE-MPs+Pb exposure groups, which indicated the skewedness to TH1 response. Meanwhile, reduced blood hemoglobin levels and increased levels of IL-4, the dominant TH2 cytokine in the Pb-only exposure group, were observed. Conclusions: Our current findings on the predominance of TH1 immune response in the PE-MPs and PE-MPs+Pb groups suggest that PE-MPs could be responsible for the predominant induction of the cellular immune changes. This finding could be used as an important landmark in research related to TH1 predominance, such as autoimmune diseases. It suggests that additional research on immune modulation using longer exposure durations or the same exposure route is required to elucidate stronger findings.

In Vitro Properties and Biodistribution of Tc-99m and Re-188 Labeled Monoclonal Antibody CEA79.4 (Re-188과 Tc-99m 표지 단일클론항체 CEA79.4의 생체외 특성과 생체내 분포)

  • Hong, Mee-Kyoung;Jeong, Jae-Min;Yeo, Jeong-Seok;Kim, Kyung-Min;Chang, Young-Soo;Lee, Yong-Jin;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul;Lee, Seung-Jin
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.6
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    • pp.516-524
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    • 1998
  • Purpose: Radiolabeled CEA79.4 antibody has a possibility to be used in radioimmunoscintigraphy or radioimmunotherapy of cancer. We investigated the in vitro properties and biodistribution of CEA79.4 antibody labeled with Re-188 or Tc-99m. Materials and Methods: CEA79.4 was reduced by 2-mercaptoethanol to produce-SH residue, and was labeled with Re-188 or Tc-99m. For direct labeling of Tc-99m, methylene-diphosphonate was used as transchelating agent. CEA79.4 in 50 mM Acetate Buffered Saline (ABS, pH 5.3) was labeled with Re-188, using stannous tartrate as reducing agent. In order to measure immunoreactivity and the affinity constant of radiolabeled antibody, cell binding assay and Scatchard analysis using human colon cancer cells SNU-C4, were performed. Biodistribution study of labeled CEA79.4 was carried out at 1, 14 and 24 hr in ICR mice. Results: Labeling efficiencies of Tc-99m and Re-188 labeled antibodies were $92.4{\pm}5.9%$ and $84.7{\pm}4.6%$, respectively, In vitro stability of Tc-99m-CEA79.4 in human serum was higher than Re-188-CEA79.4. Immunoreactivity and affinity constant of Tc-99m-CEA79.4 were 59.2% and $6.59{\times}10^9\;M^{-1}$, respectively, while those of Re-188-CEA79.4 were 41.6% and $4.2{\times}10^9\;M^{-1}$, respectively. After 24 hr of administrations of Re-188 and Tc-99m labeled antibody, the remaining antibodies in blood were 6.32 and 9.35% ID/g respectively. The biodistribution of each labeled antibody in other organs was similar because they did not accumulate in non-targeted organs. Conclusion: In vitro properties and biodistribution of Re-188-CEA79.4 were similar to those of Tc-99m-CEA79.4. It appears that Re-188-CEA79.4 can be used as a suitable agent for radioimmunotheraphy.

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Depression of Immune Response by Newcastle Disease Virus Infection (Newcastle병(病) 바이러스감염(感染)에 의(依)한 면역반응억제(免疫反應抑制))

  • Kim, Hwan-Jong;Ha, Tai-You
    • The Journal of the Korean Society for Microbiology
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    • v.14 no.1
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    • pp.79-87
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    • 1979
  • The immunosuppressive activity of newcastle disease virus(NDV) and some possible role of interferon(C-IF) in viral suppression of immune response were evaluated by SRBC-induced delayed-type hypersensitivity(DTH), rosette formation in spleen cells, number of lymphocytes in peripheral blood, hemagglutinin and hemolysin response to SRBC in ICR mice sensitized with SRBC. When NDV was inoculated before or after sensitization of mouse with SRBC, virus caused a marked inhibition of DTH, and its depressive effect was dependent on the time of virus inoculation in relation to SRBC sensitization or challenge. Rosette formation of spleen cells was significantly reduced by NDV infection. The degree of the depression of rosette formation was more prominent in mice inoculated before sensitization than after sensitization and could be related to the amount of serum interferon induced by the virus. Humoral response to SRBC of virus infected mouse was significantly depressed when NDV was inoculated 24 or 48 hours before sensitization. However, there was no difference in the response when the virus was inoculated 9 hour before and at the same time of sensitization or even after that. Lymphocytes in peripheral blood of mice were markedly diminished in numbers when NDV was inoculated 48 and 24 hour before sensitization with SRBC, but they were slightly augmented when the virus was inoculated 9 hour before and at the same time of sensitization. When UV-inactivated or heat-inactivated NDV was injected to the mouse at the same time of sensitization with SRBC, DTH and rosette formation of spleen cells were slightly depressed. DTH and rosette formation in mice treated with crude-IF were generally depressed as com pared with those of control mice. These studies suggest that the NDV causes a significant depression of cell-mediated immunity, whereas the humoral immune response is not inhibited markedly, and that the depression of immune response by NDV infection may be caused by interferon produced by NDV and direct viral activity.

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