• Title/Summary/Keyword: Mouse Size

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Accumulation of Lipid Including Cholesterol in Murine Hepatocytes with Hyperlipidemia Induced by Triton WR-1339 (Triton WR-1339 주사에 의한 고지혈증 유발시 간세포내 지방 축적에 관한 형태학적 연구)

  • Park, In-Sick;Ahn, Sang-Hyun;Kim, Jin-Taek
    • The Journal of Dong Guk Oriental Medicine
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    • v.6 no.1
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    • pp.107-115
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    • 1997
  • Hapatic tissues of ICR mouse intraperitoneally injected with Triton WR-1339 were observed to investigate the morphologic change of liver by destruction of lipid metabolism as hyperlipidemia. The hepatic tissues were obtained at hour-24, 48, and 72 after triton injection that were fixed in fromol-calcium solution and were cryocut. These tissues were stained by H&E for general morphology, sudan black B for lipid, and perchloric acid-naphthoquinone method for cholesterol. The increase of hepatocyte having meshlike cytoplasm were shown in all hepatic lobules after triton injection and the hepatic plates were disappeared in the region aggregated meshlike hepatocyte. The number of blue black colored lipid drop and dark green colored asterisk shaped cholesterol particle in hepatic cytoplasm were increased than the saline injected mouse and the size of lipid drop was enlarged. As results indicated that the lipid metabolism were destructed by triton injection, subsequently hepatocyes accumulated with lipid including cholesterol.

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External Application of Apo-9'-fucoxanthinone, Isolated from Sargassum muticum, Suppresses Inflammatory Responses in a Mouse Model of Atopic Dermatitis

  • Han, Sang-Chul;Kang, Na-Jin;Yoon, Weon-Jong;Kim, Sejin;Na, Min-Chull;Koh, Young-Sang;Hyun, Jin-Won;Lee, Nam-Ho;Ko, Mi-Hee;Kang, Hee-Kyoung;Yoo, Eun-Sook
    • Toxicological Research
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    • v.32 no.2
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    • pp.109-114
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    • 2016
  • Allergic skin inflammation such as atopic dermatitis is characterized by skin barrier dysfunction, edema, and infiltration with various inflammatory cells. The anti-inflammatory effects of Apo-9'-fucoxanthinone, isolated from Sargassum muticum, have been described in many diseases, but the mechanism by which it modulates the immune system is poorly understood. In this study, the ability of Apo-9'-fucoxanthinone to suppress allergic reactions was investigated using a mouse model of atopic dermatitis. The Apo-9'-fucoxanthinone-treated group showed significantly decreased immunoglobulin E in serum. Also, Apo-9'-fucoxanthinone treatment resulted in a smaller lymph node size with reduced the thickness and length compared to the induction group. In addition, Apo-9'-fucoxanthinone inhibited the expression of interleukin-4, interferon-gamma and tumor necrosis factor-alpha by phorbol 12-myristate 13-acetate and ionomycin-stimulated lymphocytes. These results suggest that Apo-9'-fucoxanthinone may be a useful therapeutic strategy for treating chronic inflammatory diseases.

The Inhibitory Effects of Bimanbang-1(肥滿1號方) on the Obese-mouse Fed High-fat Diet (비만1호방(肥滿1號方)이 고지방식이(高脂肪食餌)로 비만(肥滿) 유도(誘導)된 백서(白鼠)에 미치는 영향)

  • Shin, Hong-Jung;Yoon, Il-Ji
    • The Journal of Korean Medicine
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    • v.29 no.2
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    • pp.116-132
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    • 2008
  • Objective: This study investigated the effects of Bimanbang-1 (肥滿1號方: here in after referred to BMB-1) on the obese gene and obese inhibitory. Methods: C57BL/6 mice were induced by high fat diet. Mice were divided into three groups (normal, high fat diet with control, high fat diet with BMB-1 extract) and fed for 15 weeks. Items of this experimental study are as follows: body weight change, final body weight, the weight change of the adipocytes in body, the level change of LFT, NEFA and creatinine, the expression of ${\beta}3AR$ and leptin gene in primary adipocytes, the production change of leptin in primary adipocytes, the expression of ${\beta}3AR$, leptin and $TNF-{\alpha}$ in adipocytes tissue. Result: 1. All experimental groups showedthat the weight change decreased considerably and the high density group showedthat the final weight decreased considerably. 2. The high density group showed that the amount of the adipocyte in weight decreased considerably. 3. All experimental groups showedthat the amount of ALT decreased considerably, and AST decreased in the high density group. However, the amount of creatinine and glucose did not increase considerably. 4. All experimental groups showed that the amount of total cholesterol, LDL-cholesterol, triglyceride, and NEFA decreased, and HDL-cholesterol increased considerably. 5. The high density groups showedthat the amount of leptin decreased considerably. 6. All experimental groups showed that the revelation of ${\beta}3AR$ in primary adipose cell and 3T3-L1 cell increased considerably, and that the revelation of leptin and $TNF-{\alpha}$ in primary adipose cellsand 3T3-L1 cells decreased considerably. 7. All experimental groups showed that the size of adipocyte in adipocytes tissue decreased. 8. All experimental groups showed that the adipose vacuoles in liver tissue decreased considerably. Conclusion: The findings suggest that Bimanbang-1 causes weight loss and histological change, thus it may be effective to treat obesity.

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Expression of Human Serum Albumin in Milk of Transgenic Mice Using Goat β-casein/Human Serum Albumin Fusion Gene

  • Wu, H.T.;Chou, C.K.;Huang, M.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.743-749
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    • 2004
  • The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

Combination Therapy for Gliomas Using Temozolomide and Interferon-Beta Secreting Human Bone Marrow Derived Mesenchymal Stem Cells

  • Park, Jae-Hyun;Ryu, Chung Heon;Kim, Mi Jin;Jeun, Sin-Soo
    • Journal of Korean Neurosurgical Society
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    • v.57 no.5
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    • pp.323-328
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    • 2015
  • Objective : Malignant gliomas are the most common primary tumors of the central nervous system and the prognosis of patients with gliomas is poor. The combination of interferon-bata (IFN-${\beta}$) and temozolomide (TMZ) has shown significant additive antitumor effects in human glioma xenograft models. Considering that the poor survival of patients with human malignant gliomas relates partly to the inability to deliver therapeutic agents to the tumor, the tropism of human bone marrow-derived mesenchymal stem cells (MSC) for malignant gliomas can be exploited to therapeutic advantages. We investigated the combination effects of TMZ and MSCs that secrete IFN-${\beta}$ on gliomas. Methods : We engineered human MSCs to secret mouse IFN-${\beta}$ (MSC-IFN-${\beta}$) via adenoviral transduction and confirmed their secretory capacity using enzyme-linked immunosorbent assays. In vitro and in vivo experiments were performed to determine the effects of the combined TMZ and MSC-IFN-${\beta}$ treatment. Results : In vitro, the combination of MSC-IFN-${\beta}$ and TMZ showed significantly enhanced antitumor effects in GL26 mouse glioma cells. In vivo, the combined MSC-IFN-${\beta}$ and TMZ therapy significantly reduced the tumor size and improved the survival rates compared to each treatment alone. Conclusion : These results suggest that MSCs can be used as an effective delivery vehicle so that the combination of MSC-IFN-${\beta}$ and TMZ could be considered as a new option for the treatment of malignant gliomas.

Development of Bioavailability Enhancement System for the Skin Permeation Promotion of Psolarea corylifolia Extract (보골지 추출물의 피부 투과 촉진 시스템 개발)

  • Cho, Young-Ho;Ahn, Ghe-Whan;Yang, Seung-Won;Cho, Kwan-Hyun;Kim, Sang-Won;Baek, Ki-Myoung;Lee, Gye-Won
    • KSBB Journal
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    • v.26 no.6
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    • pp.505-512
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    • 2011
  • Psolarea corylifolia extract that contains bakuchiol is known to have anti-microbial, anti-inflammatory and anti-scarring effects. In this study, a vesicles such as liposome, niosome, and transfersome were produced to encapsulate P. corylifolia extract and measured their stability and physiochemical property. The skin permeation and partitioning of P. corylifolia extract in the vesicles were elucidated in nude mouse skin by using Franz diffusion cells after topical application for 24 h. After storage at 25, 40, $70^{\circ}C$, and light, the stability of bakuchiol incorporated into the vesicles was maintained for 30 days. The optimal concentration of P. corylifolia extract entrapped into the vesicles was found to be 5~10%. From the physicochemical studies, after storage at 4, 25, and $40^{\circ}C$, the viscosity and particle size of the vesicles remained in 30~80 cP and the nanosize range for 6 months, respectively. From the permeation experiments, niosome showed a higher amount of bakuchiol permeated through the mouse skin compared to liposome and transfersome after 24 h. From these results, niosome and transfersome could be a good bioavailability enhancement system (BAES) for P. corylifolia extract to improve the skin permeation and stability.

Ginsenoside Rd alleviates mouse acute renal ischemia/reperfusion injury by modulating macrophage phenotype

  • Ren, Kaixi;Jin, Chao;Ma, Pengfei;Ren, Qinyou;Jia, Zhansheng;Zhu, Daocheng
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.196-202
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    • 2016
  • Background: Ginsenoside Rd (GSRd), a main component of the root of Panax ginseng, exhibits anti-inflammation functions and decreases infarct size in many injuries and ischemia diseases such as focal cerebral ischemia. M1 Macrophages are regarded as one of the key inflammatory cells having functions for disease progression. Methods: To investigate the effect of GSRd on renal ischemia/reperfusion injury (IRI) and macrophage functional status, and their regulatory role on mouse polarized macrophages in vitro, GSRd (10-100 mg/kg) and vehicle were applied to mice 30 min before renal IRI modeling. Renal functions were reflected by blood serum creatinine and blood urea nitrogen level and histopathological examination. M1 polarized macrophages infiltration was identified by flow cytometry analysis and immunofluorescence staining with $CD11b^+$, $iNOS^+$/interleukin-12/tumor necrosis factor-${\alpha}$ labeling. For the in vitro study, GSRd ($10-100{\mu}g/mL$) and vehicle were added in the culture medium of M1 macrophages to assess their regulatory function on polarization phenotype. Results: In vivo data showed a protective role of GSRd at 50 mg/kg on Day 3. Serum level of serum creatinine and blood urea nitrogen significantly dropped compared with other groups. Reduced renal tissue damage and M1 macrophage infiltration showed on hematoxylin-eosin staining and flow cytometry and immunofluorescence staining confirmed this improvement. With GSRd administration, in vitro cultured M1 macrophages secreted less inflammatory cytokines such as interleukin-12 and tumor necrosis factor-${\alpha}$. Furthermore, macrophage polarization-related pancake-like morphology gradually changed along with increasing concentration of GSRd in the medium. Conclusion: These findings demonstrate that GSRd possess a protective function against renal ischemia/reperfusion injury via downregulating M1 macrophage polarization.

Cultural, Morphological and Pathological Variation in Indian Isolates of Ascochyta rabiei, the Chickpea Blight Pathogen

  • Basandrai, A.K.;Pande, S.;Kishore, G. Krishna;Crouch, J.H.;Basandrai, D.
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.207-213
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    • 2005
  • Cultural, morphological and pathogenic variation in Indian isolates of Ascochyta rabiei, the causal agent of blight of chickpea, was investigated. Fungal isolates representative of seven agroclimatic regions in north western plain zones (NWPZ) of India showed variation in colony colour as mouse gray with green hue, light mouse gray with slate gray centre and gray with dark brown centre, when grown on chickpea dextrose agar (CDA). Conidiomatal color of the isolates varied from brown to slate gray and black. The number of conidiomata and conidia formed on CDA ranged from 49.7 to 90.7 and $5.5\times10^4\;to\;3\times10^5cm^{-2}$, respectively. The size of conidiomata and conidia of A. rabiei isolates varied from $274\times232{\mu}m\;to\;156\times116{\mu}m$, and from $14.0\times6.2{\mu}m\;to\;10.7\times4.6{\mu}m$, respectively. Fourteen A. rabiei isolates from the seven agroclimatic regions of NWPZ were evaluated for their virulence on 180 chickpea genotypes in controlled environment. Cluster analysis based on the disease rating on a 1-9 scale indicated higher similarity coefficient (> 0.65) between isolates from different agroecological regions, while few isolates from the same region had less similarity. The 14 isolates were grouped into eight pathotypes at > 0.5 similarity coefficient. Sixteen genotypes were identified as probable differentials to distinguish A. rabiei isolates.

Expression Vectors for Human-mouse Chimeric Antibodies

  • Xiong, Hua;Ran, Yuliang;Xing, Jinliang;Yang, Xiangmin;Li, Yu;Chen, Zhinan
    • BMB Reports
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    • v.38 no.4
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    • pp.414-419
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    • 2005
  • The production of recombinant antibodies has been generally recognized as time-consuming and labor-intensive. The aim of our study is to construct mammalian expression vectors containing the cDNA encoding the human constant regions and murine variable regions to massively and cost-effectively produce full-length chimeric antibodies. Unique restriction sites flanking the Ig variable region were designed to allow for the replacement of variable regions generated by PCR. Western blot analysis of the chimeric antibodies revealed that the expressed products were of the predicted size, structure and specificity. The usefulness of the vectors was confirmed by construction of human-mouse chimeric antibody-HCAb which secretes murine antibody against the human colorectal cancer. Selected in medium containing gradually increasing methotrexate (MTX), clones with increased expression of the product gene can be efficiently generated. The secretion of recombinant chimeric antibody-HCAb yielded $30\;pg\;cell^{-1}\;day^{-1}$ at $10^{-6}\;M$ MTX. With this high-level expression from pools, the convenient and rapid production of over 100 milligram amounts per liter of recombinant antibodies may be achieved, which indicates the significant roles of pYR-GCEVH and pYR-GCEVL in the production of chimeric antibodies.

Effects of Kaempferol on Lippolysaccharide-induced Inflammation in Mouse Brain (Kaempferol이 LPS로 유도된 생쥐 중추신경계 염증에 미치는 영향)

  • Lee, Hung-Gi;Kim, Do-Hoon;Kim, Youn-Sub
    • The Korea Journal of Herbology
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    • v.30 no.1
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    • pp.77-84
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    • 2015
  • Objectives : Brain inflammation early activates the microglia and activated microglia secrete a variety of pro-inflammatory cytokines. Kaempferol, which is a flavonoid in Cuscutae Semen, shows a wide range of physiological activities, including neurons protection and anti-inflammatory actions through inhibition of pro-inflammatory mediators. The present study examined the modulatory effect of kaempferol on cytokines [tumor necrosis factor- alpha ($TNF-{\alpha}$), interleukin-1beta ($IL-1{\beta}$) and interleukin-6 (IL-6)] and cyclooxygenase-2 (COX-2) mRNA expression and microglia activation in the brain tissue of the mouse. Methods : Kaempferol was administered orally three doses of 10, 20 and 30 mg/kg respectively, once 1 hour before the lippolysaccharide(LPS) (3 mg/kg, i.p.) injection. Brain tissue was removed at 4 hours after LPS injection. Cytokines and COX-2 mRNA expression in the brain tissue was measured by the quantitative real-time polymerase chain reaction (PCR) method. Iba1 expression was calculated by western blotting method. Microglia was observed with immunohistochemistry. Immunohistochemistry stained microglia was analyzed by using ImageJ software. Results : Kaempferol 20 and 30 mg/kg was significantly attenuated the expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 mRNA. Kaempfrol 10, 20 and 30 mg/kg significantly attenuated COX-2 mRNA expression in the brain tissue. Kaempferol 30 mg/kg significantly suppressed the increase of Iba1 protein expression by LPS. Kaempferol 30 mg/kg significantly decreased the number of microglia in the cerebral cortex and the number and cell size of microglia in the hypothalamic region and the area percentage of ionized calcium binding adaptor molecule 1(Iba1)-expressed microglia in the hippocampus. Conclusions : This results indicate that kaempferol plays an anti-inflammatory role in the brain.