• Title/Summary/Keyword: Complete Freund's Adjuvant

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A Study on Antigenicity of G009, a Polysaccharide Isolated from Gandoderma lucidum IY009 in Mice and Guinea pigs (영지의 단백다당체 G009의 마우스와 기니픽에 있어서의 항원성에 관한 연구)

  • Park, Jong-Il;Jeong, Taw-Cheon;Cha, Shin-Woo;Shin, Ho-Chul;Jeong, Hoon;Kim, Su-Ung;Han, Sang-Seop
    • Biomolecules & Therapeutics
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    • v.4 no.1
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    • pp.1-6
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    • 1996
  • In the present study, the antigenic potential of G009, a polysaccharide isolated from Ganoderma lucidum IY009, was determined in BALB/C mice and Hartley guinea pigs. Antigenicity tests, including passive cutaneous anaphylaxis (PCA), active systemic anaphylaxis (ASA) and indirect hemagglutination test (IHA) were performed according to the established guidelines of National Institute of Safety Research. The results were as follows: 1. Mice showed no production of antibodies against G009 sensitized with an adjuvant, aluminum hydroxide gel (alum), when judged by the heterologous PCA test in rats. Meanwhile, antibodies against ovalbumin (OVA) sensitized with alum were clearly detected. 2. In the studies with guinea pigs, both the sensitization of G009 alone and of G009 with complete Freund's adjutant (CFA) did not produce positive reactions in homologous PCA. In the case of ASA, however, G009 alone and G009 with CFA produced positive reactions. 3. No G009 specific reaction was observed in an IHA assay using sera isolated from G009 sensitized mice. These findings suggest that G009 have no antigenicity potential in mice but may have weak antigenicity in guinea pjgs.

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Effects of the He-Ne IR Laser Irradiation and Neuromuscular Electrical Stimulation on the Change of the Serum Biochemical Components in the Experimentally Induced Muscle Injured Rats

  • Kim, Jong-Soon;Rho, Min-Hee;Kim, Jae-Young
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.223-230
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    • 2007
  • Low power laser and neuromuscular electrical stimulation have been utilized in many clinical applications for the treatment of musculoskeletal lesion. This study was to investigate the effects of low power He-Ne IR laser and neuromuscular electrical stimulation on the change of the serum biochemical components in experimentally induced muscle injured rats. The twenty Sprague-Dawley male rats were randomly assigned to the 4 groups: the normal group (n=5), the control group (n=5), the low power laser irradiation group (LLI) (n=5) and the neuromuscular electrical stimulation group (NMES) (n=5). The experimentally induced muscle injury was induced by the subcutaneous injection of a 0.1 ml Freund's Complete Adjuvant into the right hind paw and calf muscles in control, LLI and NMES groups. The LLI group was irradiated with He-Ne IR laser for 5 minutes every day during 10 days. Also, the NMES group was electrically stimulated 15 minutes every day during 10 days. The normal and control groups were not received with any therapeutic intervention. The authors performed tests which were the concentration of the serum biochemical components to detect the effects of therapeutic interventions. The results were as follows: 1. The level of the aspartate aminotransferase (AST) was significantly decreased in the LLI and NMES groups compare to the control group. 2. The level of the serum lactate dehydrogenase (LDH) was significantly decreased in the LLI and NMES groups compare to the control group. 3. The level of the serum creatine kinase (CK) was significantly decreased in the LLI and NMES groups compare to the control group. From the results, we could come to the conclusion that low power laser and neuromuscular electrical stimulation could be accelerating healing processes of skeletal muscle injury and further clinical work will be required.

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Activation of spinal Serotonergic Receptor Contributes to Electroacupuncture Analgesia in Rat with Chronic Pain (만성통증이 유발된 흰쥐에서 관찰된 침진통효과의 세로토닌성 기전)

  • Park Dong-Suk;Shin Hong-Kee;Lee Kyung-Hee
    • The Journal of Korean Medicine
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    • v.26 no.3 s.63
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    • pp.239-248
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    • 2005
  • Objectives : Electroacupuncture (EA)-induced analgesia has been known to be mediated through the activation of opioid, noradrenergic and serotonergic receptors. However, little study on serotonergic mechanism has been performed in an animal model of chronic pain. The present study was designed to elucidate the type of serotonergic receptors responsible for EA analgesia in the chronic pain model. Methods : In rats with complete Freund's: adjuvant-induced inflammation and spinal nerve injury, spinal wide dynamic range (WDR) cell responses to graded electrical stimulation of afferent C fiber were recorded before and after spinal application of selective 5-hydroxytryptamine (5-HT) receptor antagonists. EA stimulation (2Hz, 0.5msec, 3mA) was applied to the contralateral Zusanli point for 30 min. Results : In both models of chronic pain, WDR cell responses were greatly inhibited after EA stimulation. EA-induced inhibition of WDR celt responses was significantly attenuated by spinal application of non-selective 5-HT receptor antagonist, dihydroergocristine Of 5-HT receptor antagonists tested, 5-HT1A (WAY 100635) and 5-HT2 (LY53857) receptor antagonists strongly reduced an ability of EA stimulation to inhibit WDR cell responses. However, 5-HT1B (GR55562) and 5-HT3 (LY278584) receptor antagonists also had weak but significant blocking action on EA-induced inhibitory effect on chronic pain. Conclusions : Dorsal hem cell responses, afferent C fiber stimulation, chronic pain, electroacupuncture, serotonergic receptors.

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The effect of Bee Venom on type II Collagen-induced Arthritis (봉독약침이 제2형 콜라겐유도 관절염에 미치는 영향)

  • Kim, Tae-Woo;Choi, Do-Young;Lee, Jae-Dong
    • Journal of Acupuncture Research
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    • v.19 no.2
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    • pp.92-104
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    • 2002
  • Objective : Bee venom (BV) has traditionally been used in Oriental medicine to relieve pain and to treat inflammatory disease such as rheumatoid arthritis (RA). Autoimmunity to type II collagen (CII) may involve in the pathogenesis of RA. This study was performed to evaluate the effect of BV on type II collagen induced arthritis (CIA) with the naked eye, a immunohistochemical method and the examination of histology. Method : Male mice were immunized by subcutaneously injection of an $200{\mu}g$ emulsion mixed with bovine CII and complete Freund's adjuvant (CFA) twice for two weeks. In the control group, normal saline was injected, and in the experimental group, BV was applied. Result : The incidence of arthritis, the mean arthritis index and the number of the arthritic limbs of the BV group were all significantly lower than those of the control group. Among the pro-inflammatory cytokines, the production of $TNF-{\alpha}$ in the BV group was also suppressed compared with the control group, but $IL-1{\beta}$ was not. The examination on the histopathology of joints of CIA mice showed the effect of Bee Venom Herbal Acupuncture on the arthritis. Conculusion : Treatment with BV resulted in inhibition of development of arthritis and immune responses to CII.

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Morphological Changes of Bones and Joints with Rheumatoid Arthritis and Osteoarthritis

  • Hong, Yun-Kyung;Javaregowda, Palaksha Kanive;Lee, Sang-Kil;Lee, Sang-Rae;Chang, Kyu-Tae;Hong, Yong-Geun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.143-149
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    • 2011
  • Arthritis is a common disease in aged people, and is clinically divided into rheumatoid arthritis (RA) and osteoarthritis (OA). Although common symptoms such as pain are present, the underlying pathological mechanisms are slightly different. Therefore, the objectives of the present study were to compare joint damage induced by RA and OA by analyzing the major morphological and molecular differences, and to propose a suitable therapeutic intervention based on the pathophysiological conditions of bones and joints. For the RA animal model, 8-week-old DBA1/J mice were immunized with bovine type II collagen emulsified in complete Freund's adjuvant (CFA). Normal C57BL/6 mice (over 2 years of age) were used for OA. The clinical arthritis score was calculated using a subjective scoring system, and paw thicknesses were measured using calipers. The serum TNF ${\alpha}$ level was analyzed using an ELISA kit. Micro-CT was used to identify pathological characteristics and morphological changes. In collagen-induced RA mice, there were increased ankle joint volumes and clinical scores (p<0.01). The concentration of TNF ${\alpha}$ was significantly increased from 3 to 7 weeks after immunization. Micro-CT images showed trabecular bone destruction, pannus formation, and subchondral region destruction in RA mice. OA among aged mice showed narrowed joint spaces and breakdown of articular cartilage. This study suggests that a careful therapeutic intervention between RA and OA is required, and it should be based on morphological alteration of bone and joint.

A Study on the Effects of Chungyeolmaksungbang on Mouse Model of Membranous Nephropathy Induced by Cationic Bovine Serum Albumin (청열막성방(淸熱膜性方)이 Cationized Bovine Serum Albumin투여로 유발된 Mouse의 Membranous Nephropathy에 미치는 영향)

  • Choi, In-Gu;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.104-116
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    • 2008
  • Objective : Membranous nephropathy (MN) is the most common cause of adult nephrotic syndrome worldwide. MN has been defined as granular subepithelial deposition of IgG immune complexes along the glomerular basement membrane (GBM). We aimed to identify the effects of Chungyeolmaksungbang (CYMSB) treatment on cBSA-induced in MN mouse model. Methods : The effect of Chungyeolmaksungbang treatment was studied on the morphology and protein excretion in the cationized bovine serum albumin (cBSA)induced mouse chronic serum sickness nephritis model. One group of mice was given intra-peritoneal (i.p.) immunizing doses of cBSA and complete Freund's adjuvant. One week later, these animals began a single i.p. injection of cBSA for 4 weeks. A second group followed the same injection protocol, but was given CYMSB p.o. Results : Proteinuria significantly was decreased and serum albumin was increased in the group treated with cBSA and CYMSB extract compared with the control. Serum BUN was significantly decreased on CYMSB compared with control. CD3e+/CD19 cells ratio of peripheral blood was decreased and CD4+/CD8 cells was increased. Level of $IL-1{\beta}$ was significantly decreased, and $IFN-{\gamma}$ was significantly increased. Concentration of IgG and IgM was significantly decreased compared with control. Thickness of GBM was decreased on histological analysis of kidney. Deposition of CD4 and CD8 was decreased on immunohistochemical staining of kidney. Conclusions : We conclude that CYMSB treatment may could be a useful remedy agents for treating the MN with cBSA.

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The Radio-Immunoassay Method for Ginsenoside $Rg_1$ of Korean Ginseng

  • Han, Byung-Hoon;Han, Yong-Nam;Sankawa, Ushio;Akyama, Minko;Kawashima, Koichiro
    • Korean Journal of Pharmacognosy
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    • v.11 no.3_4 s.43
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    • pp.133-140
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    • 1980
  • In order to develop the radio-immunoassay procedure for the ginsenoside $Rg_1$ we prepared the $Rg_1-BSA$ conjugate and $Rg_1-tyramine$ conjugate by condensing the $Rg_1-azide$, which was prepared by a series of six step chemical modification of the $Rg_1-side$ chain, with bovine serum albumin(BSA) or with tyramine. Rabbits were immunized by repeated injection of $Rg_1-BSA$ conjugate with Freund's Complete Adjuvant for 5 month long to obtain very potent $anti-Rg_1$ serum. The radio-labelled haptene was prepared by direct radio-iodination $(125_J)$ of $Rg_1-tyramine$ according to the chloramine-T method. The radio-immunoassay procedure was successfully furnished by using DCC method (dextran coated charcoal) and the anti-body titer of the anti-serum was found as being $1600{\sim}3200$ by using 15000cpm tracer per test. Calibration test using non-labelled $Rg_1$ showed linear competetive binding response in the $(8-300){\times}34pg$. range of non-labelled $Rg_1$. The cross reaction test using 19 ginsenoside analogues enabled us a full structure-activity analysis on the antigen-antibody reaction that the anti-body in the serum would recognize the full structure of ginsenoside $Rg_1$ except the side chain moiety.

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Antigenicity test of cis-Malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolane]platinum(II)(SKI 2053R) in Guinea Pigs and Mice (기니픽 및 마우스에서 cis-Malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolane]platinum(II)(SKI 2053R)의 항원성시험)

  • Lee, Yong-Soon;Kang, Kyung-Sun;Shin, Dong-Jin;Kim, Hyoung-Ook;Cho, Jae-Jin;Kim, Bae-Hwan;Nam, Ki-Hoan;Seo, Kwang-Won
    • Toxicological Research
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    • v.8 no.2
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    • pp.255-263
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    • 1992
  • SKI 2053R and SKI 2053R-human serum albumin(HSA) mixture were examined for their antigenicity in Hartley guinea pigs as well as C57BL/6 mice in comparison with distilled water (DW), HSA and DW-HSA conjugate. Several antigenicity tests, including acitive systemic anaphylaxis(ASA), passive systemic anaphylaxis (PSA), passive cutaneous anaphylaxis (PCA) and indirect hamagglutination test (IHA), were performed according to the Established Regulations of National Institute of Safety Research. The results were as follows: 1. When guinea pigs were sensitized with SKI2053R or SKI2053R-HSA emulsified with complete Freund's adjuvant(CFA), these animals showed negative reactions in ASA and PSA. 2.No blue spot was observed on the back skin of guinea pigs in the PCA test. 3. Sera from guinea pigs revealed a negative reaction in IHA. 4.Guinea pigs were sensitized by HSA emulsified with CFA as a positive control, and these animals showed positive reactions in ASA, PSA, PCA, and IHA. As shown above, SKI2053R was considered to possess neither antigenic, nor haptenic properties, and confirmed not to have the haemagglutinating activity.

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Study on Peripheral Mechanism and Opioid Receptors Implicated in Electroacupunture-induced Inbibition of Chronic Pain (만성통증을 억제하는 전침효과의 말초성 기전과 아편양물질수용기에 관한 연구)

  • 신홍기;이서은;박동석
    • The Journal of Korean Medicine
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    • v.24 no.3
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    • pp.108-117
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    • 2003
  • Objective : The central opioid mechanism of acupuncture analgesia has been fairly well documented in acute behavioral experiments, but little electrophysiological study has been performed on the peripheral mechanism and subtypes of opioid receptors responsible for acupuncture-induced antinociception in chronic animal models. In the present electrophysiological experiment, we studied the peripheral mechanism and opioid receptor subtypes which Were implicated in electroacupuncture-induced antinociception in the rat with chronic inflammatory and neurogenic pain. Methods : In the rat with complete Freund's adjuvant-induced inflammation and spinal nerve injury, dorsal horn cell responses to afferent C fiber stimulation were recorded before and after electroacupuncture (EA) stimulation applied to the contralateral Zusanli point for 30 minutes. Also studied Were the effects of specific opioid receptor antagonists and naloxone methiodide, which can not cross the blood-brain barrier, on EA-induced inhibitory action. Results : EA-induced inhibitory action was significantly attenuated by naloxone methiodide, suggesting that EA-induced inhibition was mediated through peripheral mechanism. Pretreatment, but not posttreatment of naltrexone and spinal application significantly blocked EA-induced inhibitory actions. In inflammatory and neurogenic pain models, ${\mu}-$ and ${\delta}-opioid$ receptor antagonists (${\beta}-funaltrexamine$ & naltrindole) significantly reduced EA-induced inhibitory action, but ${\kappa}-opioid$ receptor antagonist had weak inhibitory effect on EA-induced antinociception. Conclusion : These results suggest that 2Hz EA-stimulation induced antinoeiceptive action is mediated through peripheral as well as central mechanism, and mainly through ${\mu}-$ and ${\delta}-opioid$ receptors.

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Expression of galectin-3 in the spinal cords of Lewis rats andNOD mice with experimental autoimmune encephalomyelitis (자기면역성뇌척수염 척수조직에서 galection-3의 발현)

  • Kim, Heechul;Joo, Hong-Gu;Moon, Changjong;Ahn, Meejung;Jee, Youngheun;Lim, Yoon-kyu;Koh, Chang-Sung;Shin, Taekyun
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.349-355
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    • 2004
  • The aim of this study was to evaluate the expression of galectin-3, one of beta-galactoside-binding proteins, in the experimental autoimmune encephalomyelitis(EAE) model of Lewis rats or non-obese diabetic (NOD) mice. Western blot analysis showed that galectin-3 was weakly expressed in the spinal cords of complete Freund's adjuvant(CFA) immunized control rats. In EAE, however, galectin-3 expression was significantly increased at the peak stage(days 14 post-immunization), while it was decreased slightly at the recovery stage(day 21 post-immunization). Immunohistochemical analysis showed that galectin-3 was detected in some macrophages in demyelinating lesions of NOD mice, while galectin-3 was immunoreacted in some inflammatory cells in the perivascular cuffing in rat EAE lesions. Collectively, it is postulated that the expression of galectin-3 is significantly increased in response to neuroimmunological stimulation in the central nervous system, whereas it is weak in normal rats and mice.