• Title/Summary/Keyword: mice (BALB/c)

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ppGalNAc T1 as a Potential Novel Marker for Human Bladder Cancer

  • Ding, Ming-Xia;Wang, Hai-Feng;Wang, Jian-Song;Zhan, Hui;Zuo, Yi-Gang;Yang, De-Lin;Liu, Jing-Yu;Wang, Wei;Ke, Chang-Xing;Yan, Ru-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5653-5657
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    • 2012
  • Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.

Antioxidant and Antipruritic Activities of Ethyl Acetate Fraction from Diospyros lotus Leaves (고욤(Diospyros lotus)잎 유래 Ethyl Acetate 분획물의 항산화 및 항가려움 활성)

  • Jeon, In Hwa;Kang, Hyun Ju;Kim, Sang Jun;Jeong, Seung Il;Lee, Hyun-Seo;Jang, Seon Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1635-1641
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    • 2014
  • Diospyros lotus has been cultivated for its edible fruits, which are considered to have medicinal importance. The aim of this study was to evaluate the antioxidant and antipruritic activities of water-soluble, methanol extract, and ethyl acetate (EA) fractions from D. lotus leaves. The EA fraction showed the lowest $IC_{50}$ vale (DPPH: $5.3{\mu}g/mL$, ABTS: $53.8{\mu}g/mL$). Therefore, we further investigated anti-inflammatory and antipruritic effects of the EA fraction. TNF-${\alpha}$ production increased by PMA plus A23187 treatment was significantly inhibited by the EA fraction in a dose-dependent manner. The EA fraction also inhibited histamine release from rat peritoneal mast cells stimulated by compound 48/80, which promotes histamine release. Furthermore, EA fraction had inhibitory effects on scratching behavior induced by compound 48/80 in Balb/c mice. These results suggest that the EA fraction from D. lotus leaves has potential as ameliorative agent against oxidative stress and pruritus-related disease.

The Complex Salmonella typhimurium Ghost Cells Play an Additional Role as an Immune Adjuvant (면역보강제로서 부가적인 역할을 가지는 복합 살모넬라 타이피무리움 고스트 세포)

  • Ha, Yeon Jo;Kim, Seung Tae;Kang, Ho Young;Gal, Sang Wan;Kim, Sam Woong
    • Journal of Life Science
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    • v.24 no.2
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    • pp.154-160
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    • 2014
  • Ghost cells have been recognized as eliciting humoral and cell-mediated immune responses and have also been predicted to play a role as an immune adjuvant. In this study, we used the intramuscular (IM) route to inject BALB/c mice with four vaccine groups constructed from Salmonella typhimurium ghost (STG) cells originating from different virulent strains and complex STG groups instead of heat-labile toxin (LT)-B, a type of adjuvant. Although the complex STG groups exhibited a response after a short delay, the groups showed final total IgG levels similar to those of the LT-B group, which encodes LT-B from pMMP300. The IgG1 response to the ${\chi}$3339 group was the highest response at 6 weeks, whereas IgG2a responses to the ${\chi}$3339 and JOL389 groups were higher at 6 and 8 weeks compared to those of the LT-B group. The response of vaginal sIgA to the LT-B group was generally higher than that of the other groups, whereas fecal sIgA to the LT-B group exhibited lower responses. Protection to virulent S. typhimurium in all groups was above 80%, which was similar to the LT-B group. Taken together, we suggest that STG complex groups can be used as an immune adjuvant instead of LT-B.

Effect of Baekhasuoyijung-Tang on Mouse T Cell Cytokines (백하수오이중탕물 추출물이 생쥐 면역세포의 시토킨 조절에 미치는 효과)

  • Kim, Tae-Gyun;Park, Sung-Min;Kang, Hee;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.754-761
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    • 2008
  • The purpose of this study was to evaluate the effect of Baekhasuoyijung-Tang(BHSYJT)on mouse T cell cytokines. The proliferation of mouse CD4 T cells under the influence of BHSYJT extract was measured. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 in various concentrations of BHSYJT extract, it increased proliferation of CD4 cells by 28% in $10{\mu}g/m{\ell}$ concentration and by 32% in $100{\mu}g/m{\ell}$ concentration. Treatment of CD4+ T cells stimulated by anti-CD3e and anti-CD28 with BHSYJT resulted in reduction of $IFN-{\gamma}$,but IL-4 levels is not changed. Oral administration of BHSYJT resulted in increase of both CD4+ and CD8+ T cell population in Balb/c mice by 11%. Oral administration of BHSYJT resulted in reduction of serum $IFN-{\gamma}$ level by 27% but, IL-4 level is not changed. CD4+ T cells under Th1/Th2 polarizing conditions for 3 days with BHSYJT resulted in decrease of $IFN-{\gamma}$ level in TH1 cells. Experimental results of this study show that BHSYJT helps to reduce secretion of $IFN-{\gamma}$ by mouse T helper cell in vitro and it had the same effect in vivo. Thus, it can be concluded that use of BHSYJT is an effective treatment for correcting immune imbalance in immune disorders and autoimmune diseases by reducing secretion of cytokine by Th1 cells.

Effect of Samsoeum on Cytokine Regulation of Mouse T cell (삼소음 알코올 추출물이 생쥐 면역세포의 시토킨 조절에 미치는 효과)

  • Ryu, Nae-Hyung;Park, Sung-Min;Kang, Hee;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.856-862
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    • 2008
  • The purpose of this study was to evaluate the effect of Samsoeum(SSE) on cytokine regulation of mouse T cells. The proliferation of mouse CD4 T cells under the influence of SSE extract was measured. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 in various concentrations of SSE extract, it increased proliferation of CD4 cells by 30% in $50{\mu}g/m{\ell}$ concentration. The proliferation of CD4 cells increased in $100{\mu}g/m{\ell}$ and $200{\mu}g/m{\ell}$. Treatment of CD4+ T cells stimulated by anti-CD3e and anti-CD28 with SSE resulted in reduction of $IFN-{\gamma}$ and IL-4 levels. SSE has dose-dependent inhibitory effect on $IFN-{\gamma}$ and decreased IL-4 by 70% at 50, $200{\mu}g/m{\ell}$. Oral administration of SSE resulted in increase of CD8+ T cell population in Balb/c mice by 8%. CD4+ T cells under Th1/Th2 polarizing conditions for 3 days with SSE resulted in decrease of $IFN-{\gamma}$ level in Th1 cells by 44% and increase of IL-4 level in Th2 cells by 60%. Experimental results of this study show that SSE induces mouse T-cell to transform into Th2, and increases T-cell population and activation.

Microvascular Contrast Image in Portal Veins of Rat using Micro-CT (마이크로 CT를 이용한 BALB/C(흰쥐) 간문맥의 미세혈관 조영 영상)

  • Lee, Sang-Ho;Lim, Cheong-Hwan;Jung, Hong-Rayng;Han, Beom-Hee;Mo, Eun-Hee;Chai, Kyu-Yun
    • The Journal of the Korea Contents Association
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    • v.10 no.9
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    • pp.259-266
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    • 2010
  • The study focuses on the value of Micro CT, a high resolution X-ray imaging device, by using it on rats to observe the overall portal vein image of the liver and the microvasculature of each lobes, visualize the 4 segmental lobes and acquire 3D image of the microvasculature through the reconstruction of sectional image data. Less of the damage to liver of the 5 mice, the device was able to separate the liver into 4 segmental lobes and displayed the 4 portal vein microvasculature in 2D. By using the 3D MIP technique, observation of the whole portal vein system microvasculature in 3D image was made possible along with each of the portal vein segment's branches until the 6th branch. Measured the size of 6branch, the average was measured at 1branch : $0.51mm{\pm}0.08$, 2 branch : $0.32mm{\pm}0.12$, 3 branch : $0.23mm{\pm}0.11$, 4 branch : $0.19mm{\pm}0.08$, 5 branch : $0.13mm{\pm}0.06$, 6 branch : $70.5{\mu}m{\pm}14.1$. The 3D image and the images of the microvasculatures in the result of study proved that the Micro-CT can be considered many useful device in obtaining high resolution images.

An Experimental Study on Effects of Distilled White-ginseng Herbal Acupuncture on A549 human ephithelial lung cancer cell in vitro and implanted Sarcoma-180 in vivo (A549 폐암세포와 Sarcoma-180 복강암에 대한 인삼(人蔘) 증류약침(蒸溜藥鍼)의 영향에 관(關)한 실험적(實驗的) 연구(硏究))

  • We, Jong-Seong;Kwon, Ki-Rok;Park, Hee-Soo
    • Journal of Pharmacopuncture
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    • v.7 no.3
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    • pp.59-71
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    • 2004
  • Objectives : In order to investigate effects and immune improvement of distilled white-ginseng herbal extract, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled white-ginseng Herbal Acupuncture at Wisu($Bl_{21}$) and Chung-wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR. the control group and the experiment groups show significant increase. For Cox-2, both experiment groups and the normal group showed significant decrease. For Bcl-2, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups 2. For survival time, all of experiment groups didn't show significant differences. 3. IL-2 productivity using Flow cytometry, experiment group I didn't show any significance, For II-4, all of experiment groups showed slight decrease compared to the control group. 4. For IL-2 productivity using ELISA, experiment group I showed slight decrease compared to the control group, experiment group II didn't show any significance. 5. For expression of cytokine mRNA using RT-PCR, significant increase of IL-2 and IL-4 were witnessed in experiment group I compared to the control group. Significant decrease of IL-10 was shonwn in all of experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled white-ginseng Herbal Acupuncture may be further effects in anti-cancer and immune improvement if increasing concentration.

Production of A Monoclonal Antibody (MAb) Against a Thermal Stable-Soluble Protein in Mackerel and Confirmation of the Properties for the MAb (고등어 어육 중 열안정성 단백질에 특이한 단클론성 항체 개발과 특성 확인)

  • Lee, Jeong-Eun;Kim, Jeong-Sook;Chung, Duck-Hwa;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.32 no.1
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    • pp.75-81
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    • 2017
  • For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without cross-reaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

Geumeunyoungji-tang Inhibits Serum IgE Level in Mouse Dermatitis Induced by 2,4-dinitrofluorobenzene

  • Phan, Manh Hung;Jeon, Yong Deok;Kim, Min Cheol;Kim, Su Jin;Kim, Dae Seung;Kwon, Dong Yeul;Kim, Seong Hee;No, Seung Il;Kim, Hyun Jeong;Shin, Hyun Ji;Um, Jae Young;Hong, Seung Heon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.2
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    • pp.217-222
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    • 2014
  • Lonicerae Flos containing formulation, Geumeumyoungji-tang (GYT), is an herbal prescription prepared using 5 different herbal drugs, namely, Lonicerae Flos, Ganoderma, Lactucae Herba, Xanthii Fructus, and Smilacis Rhizoma. This study was focused on the investigation of the pharmacological effects of GYT on allergic reactions. As the first step of the study, GYT was administered BALB/c mice which were sensitized by 2,4-dinitrofluorobenzene (DNFB). As the result GYT ameliorated dermatitis provoked by DNFB. The serum IgE level of the DNFB sensitized-mouse was significantly decreased when GYT was administered. In order to confirm the moderating effect of this prescription on allergic reaction, GYT was pretreated to human mast cells (HMC-1) before they were stimulated by phorbol-12-myristate 13-acetate plus calcium ionophore A23187 (PMACI). GYT suppressed secretion of inflammatory cytokines, interleukin (IL)-6 and IL-8, from HMC-1. Additionally, pretreatment of GYT showed regulating effect on COX-2 expression. Collectively, these findings provide insights into the pharmacological actions of GYT as a potential agent for treatment of allergic dermatitis.

Purification of antigenic protein of sparganum by immunoaBnity chromatography using a monoclonal antibody (단세포군항체를 이용한 친화성 크로마토그래피에 의한 스파르가눔 항원의 순수분리)

  • Cho, Seung-Yull;Kang, Shin-Yong;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • v.28 no.3
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    • pp.135-142
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    • 1990
  • The quality improvement of antigen (crude saline extract) of Spirometra maptscni 1)lerocercoid (sparganum) was investigated by protein purificatioll. The crude extract was fractionated by gel filtration through Sephacryl S-300 Superfine. Its third fraction was purified by affinity chromatography using a monoclonal antibody as ligand. When observed by SDS-PAGE, the purified protein was composed of 2 bands of 36 kDa and 29 kDa which were found already as the most sensitive components in the crude extract by immunoblots with patients sera. The quality of the purified antigen was evaluated in comparison with the crude extract by ensyme-linked imnunosorbent assay (ELISA) for the specific (IgG) antibody in sera of human sparganosis, other parasitic and neurologic diseases, and normal control. When the purified antigen was used: the sensitivity was not altered but remained high (96.4%) while the specificity was increased from 86.8% to 96.9%.

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