• Title/Summary/Keyword: In-vivo ${\mu}-CT$

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Activity-guided Purification of N-benzyl-N-methyldecan-1-amine from Garlic and Its Antitumor Activity against CT-26 Colorectal Carcinoma in BALB/C Mice (활성추적분리법에 의해서 순수분리한 마늘 N-benzyl-N-methyldecan-1-amine이 CT-26 세포주 이식 BALB/C mice의 항암효과)

  • Seetharaman, Rajasekar;Choi, Seong Mi;Guo, Lu;Cui, Zheng Wei;Otgonbayar, Duuriimaa;Park, Ju Ha;Kwon, Young-Seok;Kwak, Jung Ho;Kwon, Young Hee;Min, Ji Hyun;Kang, Jum Soon;Choi, Young Whan
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1062-1070
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    • 2019
  • A components of garlic (Allium sativum) have anti-proliferative effects against various types of cancer. We aimed to investigate the capacity of garlic compounds to anti-tumor on a various cancer cell lines. Fractionation of garlic extract, guided by antiproliferative activity against human gastric cancer (AGS) cells, has resulted in the isolation of N-benzyl-N-methyldecan-1-amine (NBNMA). We investigated the effect of newly isolated NBNMA from garlic cloves on the inhibition of the growth of CT-26, AGS, HepG2, HCT-116, MCF7, B16F10, and Sarcoma-180 cells for in vitro and CT-26 colon carcinoma cells in vivo. NBNMA exhibited an antiproliferative effect in CT-26 cells by apoptotic cell death. NBNMA exhibited down-regulation of anti-apoptotic Bcl-2 proteins and up-regulation of apoptotic Bad protein expression in western blot analyses. In addition, NBNMA meagre activated caspase 3 and caspase 9, initiator caspases of the extrinsic and intrinsic pathways of apoptosis. NBNMA treatment at a dose of 10 mg/kg for 21 days in experimental mice implanted with tumors resulted in significant reduction of the tumor weight (43%). NBNMA exhibited both in vitro and in vivo anticancer activity. These results indicate that NBNMA has promising potential to become a novel anticancer agent from garlic cloves for the treatment of colon carcinoma cancer.

Ex vivo Morphometric Analysis of Coronary Stent using Micro-Computed Tomography (미세단층촬영기법을 이용한 관상동맥 스텐트의 동물 모델 분석)

  • Bae, In-Ho;Koh, Jeong-Tae;Lim, Kyung-Seob;Park, Dae-Sung;Kim, Jong-Min;Jeong, Myung-Ho
    • Journal of the Korean Society of Radiology
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    • v.6 no.2
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    • pp.93-98
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    • 2012
  • Micro-computed tomography (microCT) is an important tool for preclinical vascular imaging, with micron-level resolution. This non-destructive means of imaging allows for rapid collection of 2D and 3D reconstructions to visualize specimens prior to destructive analysis such as pathological analysis. Objectives. The aim of this study was to suggest a method for ex vivo, postmortem examination of stented arterial segments with microCT. And ex vivo evaluation of stents such as bare metal or drug eluting stents on in-stent restenosis (ISR) in rabbit model was performed. The bare metal stent (BMS) and drug eluting stent (DES, paclitaxel) were implanted in the left or right iliac arteries alternatively in eight New Zealand white rabbits. After 4 weeks of post-implantation, the part of iliac arteries surrounding the stent were removed carefully and processed for microCT. Prior to microCT analysis, a contrast medium was loaded to lumen of stents. All samples were subjected to an X-ray source operating at 50 kV and 200 ${\mu}A$ by using a 3D isotropic resolution. The region of interest was traced and measured by CTAN analytical software. Objects being exposed to radiation had different Hounsfield unit each other with values of approximately 1.2 at stent area, 0.12 ~ 0.17 at a contrast medium and 0 ~ 0.06 at outer area of stent. Based on above, further analyses were performed. As a result, the difference of lengths and volumes between expanded stents, which may relate to injury score in pathological analysis, was not different significantly. Moreover, ISR area of BMS was 1.6 times higher than that of DES, indicating that paclitaxel has inhibitory effect on cell proliferation and prevent infiltration of restenosis into lumen of stent. And ISR area of BMS was higher ($1.52{\pm}0.48mm^2$) than that of DES ($0.94{\pm}0.42mm^2$), indicating that paclitaxel has inhibitory effect on cell proliferation and prevent infiltration of restenosis into lumen of stent. Though it was not statistically significant, it showed that the extent of neointema of mid-region of stents was relatively higher than that of anterior and posterior region in parts of BMS as showing cross-sectional 2-D image. suggest that microCT can be utilized as an accessorial tool for pathological analysis.

Effects of Inonotus obliquus Extracts on Immunomodulating Activity

  • Song, Kee-Cheol;Choi, Byung-Lyul;Shin, Jang-Woo;Son, Jin-Young;Yoo, Hwa-Seung;Cho, Jung-Hyo;Lee, Yeon-Weol;Son, Chang-Gue;Cho, Chong-Kwan
    • The Journal of Korean Medicine
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    • v.28 no.4
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    • pp.27-41
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    • 2007
  • Aim : This study aimed at elucidating the effects of Inonotus obliquus on anti-tumor effects in vivo and immune-based characterization of the mushroom as a potential candidate for cancer remedy. Methods : To investigate the immunomodulatory effects of Inonotus obliquus, we investigated macrophage functions and NK cell activities through the measurement of NO production of macrophage, NK cell cytotoxicity and expressions of cytokines and genes regulating immune responses, in addition to pulmonary metastasis model in vivo. Results : Inonotus obliquus showed general cytotoxicity at high concentrations over the 100 ${\mu}g/m{\ell}$ on the both of normal and cancer cell lines. Inonotus obliquus showed both inhibitory and promotive effects on pulmonary colonization of CT-26 cell depending on period or route of administration in vivo. Conclusion : From these results, it cannot be concluded that Inonotus obliquus has cancer-specific activity. Furthermore, Inonotus obliquus has the provability to show adverse effects differently according to the concentration and the method of administration.

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Clinical Application of in Vivo Dosimetry System in Radiotherapy of Pelvis (골반부 방사선 치료 환자에서 in vivo 선량측정시스템의 임상적용)

  • Kim, Bo-Kyung;Chie, Eui-Kyu;Huh, Soon-Nyung;Lee, Hyoung-Koo;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.27 no.1
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    • pp.37-49
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    • 2002
  • The accuracy of radiation dose delivery to target volume is one of the most important factors for good local control and less treatment complication. In vivo dosimetry is an essential QA procedure to confirm the radiation dose delivered to the patients. Transmission dose measurement is a useful method of in vivo dosimetry and it's advantages are non-invasiveness, simplicity and no additional efforts needed for dosimetry. In our department, in vivo dosimetry system using measurement of transmission dose was manufactured and algorithms for estimation of transmission dose were developed and tested with phantom in various conditions successfully. This system was applied in clinic to test stability, reproducibility and applicability to daily treatment and the accuracy of the algorithm. Transmission dose measurement was performed over three weeks. To test the reproducibility of this system, X-tay output was measured before daily treatment and then every hour during treatment time in reference condition(field size; $10 cm{\times} 10 cm$, 100 MU). Data of 11 patients whose pelvis were treated more than three times were analyzed. The reproducibility of the dosimetry system was acceptable with variations of measurement during each day and over 3 week period within ${\pm}2.0%$. On anterior- posterior and posterior fields, mean errors were between -5.20% and +2.20% without bone correction and between -0.62% and +3.32% with bone correction. On right and left lateral fields, mean errors were between -10.80% and +3.46% without bone correction and between -0.55% and +3.50% with bone correction. As the results, we could confirm the reproducibility and stability of our dosimetry system and its applicability in daily radiation treatment. We could also find that inhomogeneity correction for bone is essential and the estimated transmission doses are relatively accurate.

Micro-CT System for Small Animal Imaging (소동물영상을 위한 마이크로 컴퓨터단층촬영장치)

  • Nam, Ki-Yong;Kim, Kyong-Woo;Kim, Jae-Hee;Son, Hyun-Hwa;Ryu, Jeong-Hyun;Kang, Seoung-Hoon;Chon, Kwon-Su;Park, Seong-Hoon;Yoon, Kwon-Ha
    • Progress in Medical Physics
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    • v.19 no.2
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    • pp.102-112
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    • 2008
  • We developed a high-resolution micro-CT system based on rotational gantry and flat-panel detector for live mouse imaging. This system is composed primarily of an x-ray source with micro-focal spot size, a CMOS (complementary metal oxide semiconductor) flat panel detector coupled with Csl (TI) (thallium-doped cesium iodide) scintillator, a linearly moving couch, a rotational gantry coupled with positioning encoder, and a parallel processing system for image data. This system was designed to be of the gantry-rotation type which has several advantages in obtaining CT images of live mice, namely, the relative ease of minimizing the motion artifact of the mice and the capability of administering respiratory anesthesia during scanning. We evaluated the spatial resolution, image contrast, and uniformity of the CT system using CT phantoms. As the results, the spatial resolution of the system was approximately the 11.3 cycles/mm at 10% of the MTF curve, and the radiation dose to the mice was 81.5 mGy. The minimal resolving contrast was found to be less than 46 CT numbers on low-contrast phantom imaging test. We found that the image non-uniformity was approximately 70 CT numbers at a voxel size of ${\sim}55{\times}55{\times}X100\;{\mu}^3$. We present the image test results of the skull and lung, and body of the live mice.

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Suppression of Osteoporotic bone loss on the site to which low Intensity Ultrasound is Irradiated - In vivo test on BMD and Morphological Characteristics (저강도 초음파 조사 부위의 골 소실 억제 효과 -골 밀도 및 형태학적 특성 평가)

  • Ko, Chang-Yong;Seo, Dong-Hyun;Kim, Hyo-Seon;Kim, Han-Sung;Kim, Sang-Hee;Kim, Jin-Man;Kim, Ki-Won;Lim, Do-Hyung
    • Journal of Biomedical Engineering Research
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    • v.30 no.1
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    • pp.49-55
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    • 2009
  • Purpose : The aim of this study is to evaluate a possibility of clinical application for the effects of low intensity ultrasound stimulation (LIUS) in morphological characteristics (i.e., structure, bone mineral density) of bone on osteoporotic fracturesprevention. Materials and Methods : Eight virgin 14-week-old ICR mice (approximate weight 25g) were used and ovariectomized (OVX) to induce osteoporosis. Right tibia (US) for each mouse served as the LIUS (1.5MHz frequency, 1.0 kHz pulse repetition on frequency, $30mW/cm^2$ intensity, $200{\mu}s$ pulse length, and stimulation for 20 minutes a day and 5 days a week over a 6-week period). Left tibia (CON) for each mouse served as the non-stimulated controls. Structural parameters and bone mineral density ($g/cm^3$) on trabecular bone of tibiae were calculated and measured from images derived in-vivo micro computed tomography (micro-CT) at 0 week and after 6weeks. Results : The BV/TV and Tb.N in US group were significantly bigger than those in CON group. The Tb.Pf in US group, moreover, was significantly smaller than that in CON group (p<0.05).For the others structural parameters and BMD, however, there were no significant difference between US group and CON group (p>0.05). Conclusion : The LIUS might prevent bone loss and keep bone connectivity in osteoporotic bones. Therefore, the LIUS might prevent effectively the osteoporotic fractures in clinics.

Bioactive Utility of the Extracts from Rhus verniciflua Stokes (RVS) : Biological Function of the Extracts from RVS (옻나무 추출물의 생리활성 이용에 대한 연구 : 옻나무 추출물의 생물학적 기능)

  • Lim, Kye-Taek;Lee, Jeong-Chae
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.238-245
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    • 1999
  • Antioxidative effects of the water or ethanol extracts from Rhus verniciflua Stokes (RVS) were measured by protection against hydroxyl radicals in mouse brain tissue culture. In the water extracts from RVS, cell viabilities were estimated 60.0, 66.0, 72.0, 84.0 and 90.0% at addition of 1, 2, 4, 7 and $10{\mu}L$, respectively, compared with GO (20 mU/mL) alone. The cell viability in the ethanol extracts was similarly with water extracts. In the antitumor effects, the results showed that percentages of the HeLa cell death were approximately 24% for 12 hrs, 57% for 48 hrs at addition of 10%/well ethanol extracts respectively. To know inhibition of tumor growth, in vivo, mice (BALB/c) were inoculated with 0.25 mL CT-26 $(1{\times}10^6\;cells/mL)$ subcutaneously. After the generation of tumor, the results of RVS extracts (ethanol, water) injection showed generally that the tumor size in BALB/c was reduced. For physicochemical characterization of the RVS extracts, purified substances of water or ethanol extracts were analized with SDS-PAGE and ICP spectrometer. In electrophoresis, gel showed 2 bands (210, 230 KDa). The results of ICP verified that RVS extracts contain $Cu^{2+}$ in both samples. Conclusively, this substance might be a laccase which has a biological effective function, as a natural bioactive substance.

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Therapeutic Effects of Curdrania tricuspidata Leaf Extract on Osteoarthritis (골관절염 실험모델에서 꾸지뽕나무 추출물의 골관절염 억제효과 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.697-704
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    • 2013
  • The inhibitory effect of ethanol extracts from Curdrania tricuspidata leaves (CTL) on osteoarthritis was investigated in primary cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced arthritis rat model. To identify the effects of CTL 80% ethanol extracts (CTL80) and CTL 10% ethanol extracts (CTL10) against $H_2O_2$ treatment in vitro, cell survival was measured by the MTT assay. Cell survival after $H_2O_2$ treatment increased with CTL80 and CTL10 close to normal up to $300{\mu}g/mL\;H_2O_2$. The mRNA expression of matrix metalloproteinases (MMPs) was determined MMP-7 and MMP-13 (known catabolic factors), were significantly inhibited by CTL 80 and CTL10; a $200{\mu}g/mL$ dose of CTL80 especially decreased MMP-13 expression. In vivo, osteoarthritis was induced by an intra-articular injection of MIA into the knee joints of rats, then CTL80 and CTL10 orally administered daily for 35 days. After the animals were sacrificed, histological evaluations of their knee joints revealed a reduction in polymorphonuclear cell infiltration and smooth synovial lining in the CTL80-500 group. Micro-CT analysis of hind paws from CTL80-500 and CTL10 showed a protection against osteophyte formation, soft tissue swelling, and bone resorption. In conclusion, CTL ethanol extracts are effective in ameliorating joint destruction and cartilage erosion in MIA-induced rats. CTL decreases and normalizes articular cartilage through preventing extracellular matrix degradation and chondrocyte injury, and could potentially serve as a therapeutic treatment for humans.

Effects of Keonbodan (健步丹) on MIA-Induced Osteoarthritis in Rat (건보단(健步丹)이 MIA로 유도된 골관절염 Rat에 미치는 영향)

  • Lee, Eun-Jung;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.2
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    • pp.51-64
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    • 2014
  • Objectives This study was carried out to know the effects of Keonbodan (hereinafter referred to KBD) in osteoarthritis induced by Monosodium iodoacetate(hereinafter referred to MIA) on Wistar rat. Methods Osteoarthritis was induced by injection of MIA into left knee joint cavities of rat. Osteoarthritis rats were divided into 4 groups (normal (n=6), control (n=6), indomethacin (n=6), KBD (n=6) group). The control group was administered normal saline and indomethacin group was administered indomethacin (2 mg/kg). And the KBD group was administered KBD (142 mg/kg). Each groups were administered by orally for 4 weeks. This experiment were carried out in vivo. In vivo, at the end of the experiment (5 weeks after MIA injection), effects on hepatotoxicity and nephrotoxicity, cytokines in serum, arachidonic acid, osteocalcin, MMP-9, TIMP-1 and cartilage volume were evaluated. And histopathological examinations on the articular structures of knee joints were performed. Results 1. In weight-bearing measurement, level of weight was increased. 2. In order to hepatotoxicity and nephrotoxicity, ALT, AST, BUN and creatinine were tested. And there were no significant changes. 3. In serum, levels of TNF-$\alpha$, IL-$1{\beta}$ were significantly decreased. IL-6 was insignificantly decreased. 4. In serum, level of MMP-9 and TIMP-1 was decreased. 5. In serum, level of $LTB_4$, $PGE_2$ and osteocalcin was decreased. 6. In ${\mu}$CT-arthrography, the cartilage volume was greater than that of the control group. 7. The joint damage induced by osteoarthritis was lesser than the control group in histopathologic observation (H&E, Safranin-O staining). Conclusions These results demonstrated that KBD suppressed the osteoarthritis- inducing effects of MIA in rat. And further studies are required to find out more effective substance and anti-osteoarthritic mechanism in the future.

A Study on the Effect and Mechanism of Gamikyejakjimogawusul-tang Herbal Acupuncture on Induced Rheumatoid Arthritis model of DBA/1 mice (계작지모가우슬탕(桂芍知母加牛膝湯) 약침이 류마티스 관절염 생쥐에 미치는 영향)

  • Jung, Soon Hyun;Cho, Chong kwan;Kim, So Yun;Kim, Young Il
    • Journal of Haehwa Medicine
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    • v.24 no.2
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    • pp.35-57
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    • 2016
  • Objectives : The purpose of this study is to prove the effect and mechanism of Gamikyejakjimogawusul-tang(GKHA) herbal acupuncture on induced rheumatoid arthritis model of DBA/1 mice. Methods : We check effect of GKHA extract on the AST, ALT, Creatinine, BUN of serum and cell viability of GK extract in RAW 264.7 cells to test the stability of this study. In vitro, we measure total phenol contents, total flavonoid contents, DPPH free radical scavenging activity, ABTS cation radical scavenging activity of Gamikyejakjimogawusul-tang, effect of GK extract on ROS(Reactive Ooxygen Species) production to estimate a anti-oxidant capacity, and we also measure effect of GK extract on NO (Nitric Oxid), IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF production in RAW 264.7 cells to estimate a anti-inflammatory efficacy. In vivo, we compare a rheumatoid arthritis manifestation between control and experimental group and estimate a AI. Then we check effect of GKHA on the level of WBC, neutrophil, lympocyte, monocyte in the blood to see the effect of immune cells in blood. In addition we measure effect of GKHA on the level of hs-CRP, IgM, IgG, IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF in serum. We observe effects of GKHA on imaging of cartilage degeneration using micro CT-arthrography in paw hind. And we calculate effects of GKHA that reduced BV ratio, BS/BV ratio using 3D Micro-CT. Lastly we observe effects of GKHA histopathologic examination analysis. Results : 1. The toxicity on liver and kidney was disregardable and the cytotoxicity against RAW 264.7 cells was also disregardable. 1. Total phenol contents and total flavonoid contents in GK extract were in high level. 2. DPPH free radical scavenging activity and ABTS cation radical scavenging activity were increased according to concentration of GK extract 3. ROS production was significantly decreased in GK extract (at 10, $100{\mu}g/ml$). 4. NO, IL-6, TNF-${\alpha}$, MCP-1 production were significantly decreased in GK extract(at 10, $100{\mu}g/ml$). IL-17, GM-CSF production were significantly decreased in GK extract(at 1, 10, $100{\mu}g/ml$). IL-$1{\beta}$, IL-21 production were also decreased but there was no statistical significance. 5. 25x observation after H&E and M-T staining, infiltration of immune cells and subsidence of the cartilage and damage to the synovial cells were decreased. Conclusions : This study showed that GKHA extract had anti-oxidant capacity, anti-inflammatory efficacy. GKHA extract also had inhibiting effect on the process of rheumatoid arthritis and can protect joint and cartilage. So we expect that GKHA extract can be a meaningful treatment to rheumatoid arthritis patients.