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A Study on the Effect of Herbal-acupuncture with Carthami Flos at Joksamni($ST_{36}$) on Collagen-induced Arthritis in Mice (족삼리(足三里) 홍화약침(紅花藥鍼)처치가 Collagen으로 유발한 생쥐의 관절염 모델에 미치는 영향)

  • Park, Ki-Hong;Lee, Hyun
    • Journal of Acupuncture Research
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    • v.24 no.6
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    • pp.45-61
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    • 2007
  • Objectives : The purpose of this study is to observe the effects of Carthami Flos herbal-acupuncture (CF-HA) at Joksamni($ST_{36}$) on arthritis in mice induced by Collagen II. Methods : The author performed several experimental items, including arthritis evaluation, change in weight, spleen size and stenosis rate, change in cytokine level, IgG, IgM and anti-collagen II, change of immunocyte count and histological change of the CIA mouse joint. Conclusions are as follows: Results : 1. In the CF-HA, the arthritis index and rate and the incidence of arthritis were decreased as the experiment proceeded. 2. In the CF-HA, spleen swell and stenosis, joint edema and change were decreased. 3. In the CF-HA, the level of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ and $IFN-{\gamma}$ in blood serum were significantly decreased. 4. In the CF-HA, the level of IgG, IgM and anti-collagen II were decreased. 5. In the CF- HA, $IFN-{\gamma}$, $IFN-{\gamma}/IL-4$, IL-10 of the culture fluid was decreased. 6. In the CF-HA, the cell rate of $CD3e^+$ and $CD45R^+$, $CD4^+$ and $CD8^+$, $CD4^+/CD25^+$ in spleen was similar to the cell rate of the normal group. 7. In the CF-HA, the cell rate of $CD4^+/CD25^+$, $CD45R^+/CD69^+$ in a lymph node was decreased as in the normal group. 8. In the CF-HA, the cell rate of $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in joints was decreased as in the normal group. 9. In the CF-HA, the cartilage destruction and the inflammation cell growth in the H&E stain were decreased. The collagen fiber in the M&T stain were less destructed, therefore the result was similar to the normal group. Conclusions : These results suggest that CH-HA at $ST_{36}$ has an effect in controlling immune reaction and suppressing inflammation in rheumatoid arthritis therefore, the continuous flow of the following study is expected.

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Production and Breeding of Transgenic Cloned Pigs Expressing Human CD73

  • Lee, Seung-Chan;Lee, Haesun;Oh, Keon Bong;Hwang, In-Sul;Yang, Hyeon;Park, Mi-Ryung;Ock, Sun-A;Woo, Jae-Seok;Im, Gi-Sun;Hwang, Seongsoo
    • Development and Reproduction
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    • v.21 no.2
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    • pp.157-165
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    • 2017
  • One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-human-primate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were $127{\pm}18.9$. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as ${\alpha}-1,3-galactosyltransferase$ knock-out /hCD46 for xenotransplantation.

Effector Memory CD8+ and CD4+ T Cell Immunity Associated with Metabolic Syndrome in Obese Children

  • Yang, Da-Hee;Lee, Hyunju;Lee, Naeun;Shin, Min Sun;Kang, Insoo;Kang, Ki-Soo
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.24 no.4
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    • pp.377-383
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    • 2021
  • Purpose: We investigated the association of effector memory (EM) CD8+ T cell and CD4+ T cell immunity with metabolic syndrome (MS). Methods: Surface and intracellular staining of peripheral blood mononuclear cells was performed. Anti-interleukin-7 receptor-alpha (IL-7Rα) and CX3CR1 antibodies were used to stain the subsets of EM CD8+ T cells, while anti-interferon-gamma (IFN-γ), interleukin-17 (IL-17), and forkhead box P3 (FOXP3) antibodies were used for CD4+ T cell subsets. Results: Of the 47 obese children, 11 were female. Children with MS had significantly higher levels of serum insulin (34.8±13.8 vs. 16.4±6.3 µU/mL, p<0.001) and homeostasis model assessment of insulin resistance (8.9±4.1 vs. 3.9±1.5, p<0.001) than children without MS. Children with MS revealed significantly higher frequencies of IL-7Rαlow CD8+ T cells (60.1±19.1% vs. 48.4±11.5%, p=0.047) and IL-7RαlowCX3CR1+ CD8+ T cells (53.8±20.1% vs. 41.5±11.9%, p=0.036) than children without MS. As the serum triglyceride levels increased, the frequency of IL-7RαlowCX3CR1+ and IL-7RαhighCX3CR1- CD8+ T cells increased and decreased, respectively (r=0.335, p=0.014 and r=-0.350, p=0.010, respectively), in 47 children. However, no CD4+ T cell subset parameters were significantly different between children with and without MS. Conclusion: In obese children with MS, the changes in immunity due to changes in EM CD8+ T cells might be related to the morbidity of obesity.

Effect of bee pollen extract on activation of dendritic cells and induction of Th1 immune response (꿀벌 꽃가루 열수 추출물의 수지상 세포 활성화 및 Th1 반응에 미치는 효과)

  • Cho, Eun-Ji;Kim, Yi-Eun;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.444-450
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    • 2018
  • Dendritic cells (DCs) are potent antigen-presenting cells that play a pivotal role in modulating both innate and adaptive immunity. This study examined the immunomodulatory activities of hot-water extracts of bee pollen (BPW) in bone-marrow derived DCs (BMDC) and mice splenocytes. BMDCs isolated from mice were treated with 250 and $500{\mu}g/mL$ BPW for 24 h. BPW, up to $500{\mu}g/mL$, did not display any cellular toxicity against BMDCs. In fact, it functionally induced BMDC activation via augmentation of CD80, CD86, and major histocompatibility complex (MHC) class I/II expression and pro-inflammatory cytokine (tumor necrosis factor; $TNF-{\alpha}$, interleukin; IL-6, and $IL-1{\beta}$) production. Interestingly, BPW treatment significantly increased the production of interferon $(IFN)-{\gamma}$ in splenocytes, suggesting its possible contribution to Th1 polarization in immune response. Taken together, these findings suggest that BPW may regulate innate and adaptive immunity via DC activation and Th1 polarization in immune responses.

Immunoregulatory Effects of Water Extracts of Scutellariae Radix in DSS-Induced Inflammatory Bowel Disease Animal Model (DSS로 유도된 염증성 장 질환 동물 모델에서 황금 열수 추출물이 면역 조절 기능에 미치는 영향)

  • Lee, Sun-Hee;Lim, Beong-Ou;Choue, Ryo-Won
    • Journal of Nutrition and Health
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    • v.37 no.6
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    • pp.431-439
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    • 2004
  • Scutellariae Radix (Scu.), one of the immune-regulatory substances, is recognized to play the role in the metabolic process of inflammation, allergy and immunity. It has been traditionally used in the Oriental medicine to treat inflammatory bowel diseases (IBD). The purpose of this study was to evaluate the effects of water extracts of Scutellariae Radix on the spleen lymphocyte immune function in the Balb/c female mice treated with dextran sodium sulfate (DSS) to induce colitis. Water extract of Scutellariae Radix (100 mg/kg) and sulfasalazine (50 mg/kg) were administrated orally for 2 weeks of experimental period. Mice were divided into three experimental groups randomly: DSS group (5% DSS was ad libitum for 5 days) as control group, DSS + Scu. (water extracts of Scutellariae Radix for 2 weeks after 5% DSS was ad libitum for 5 days) as experimental group, and DSS + Sulfasalazine group (Sulfasalazine for 2 weeks after 5% DSS was ad libitum for 5 days) as positive control group. Levels of Ig A, Ig E, CD4$^{+}$, CD8$^{+}$, TNF-$\alpha$ and other cytokines were measured. Treatment of DSS for 5 days induced bowel inflammation and the treatment with Scu. water exteract and sulfasalazine significantly recovered the damage. The length of intestine of DSS group was significantly shorter than that of other groups. The serum and fecal concentration of Ig A of SS + Scu group was higher than those of DSS group. The contents of CD4$^{+}$ T cells was higher in the DSS + Scu. group than the other groups and CD8$^{+}$ T cells was the lowest in DSS + Sulfasalazine group. The Ig A level of cultured supernatant of spleen lymphocyte was the highest, while the Ig E level was the lowest in SS + Scu group. The concentration of TNF-$\alpha$, cytokine secreted from the Th1 cell in the supernatant spleen lymphocyte, was the highest in the DSS group and the lowest in the DSS + Scu. group. The concentration of IFN-${\gamma}$ and ll...-12 was lower in the DSS + Scu. group than those of the other groups. The concentration of IL-4 in the supernatant of spleen lymphocyte was the lowest in the DSS + Scu. group but IL-10 was not significantly different. Based on these findings, water extract of Scutellariae Radix exhibited the inhibitory effect via IL-4 production thereby inhibited the production of Ig E and strengthened immune system, and alleviated injury in DSS- induced colitis mice model.

Influence of Ionizing Radiation on Ovarian Carcinoma SKOV-3 Xenografts in Nude Mice under Hypoxic Conditions

  • Zhang, Yong-Chun;Jiang, Gang;Gao, Han;Liu, Hua-Min;Liang, Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2353-2358
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    • 2014
  • Purpose: We aimed to detect the expression of HIF-1${\alpha}$, VEGF, HPSE-1 and CD31 in SKOV3 xenografts in nude mice treated with different doses of ionizing radiation, trying to explore the possible mechanism of hypoxia and radioresistance. Methods: Nude mice bearing SKOV3 xenografts were randomly divided into 4 groups: Group A (control group, no ionizing radiation), Group B (treated with low dose of ionizing radiation: 50cGy), Group C (treated with high dose of ionizing radiation: 300cGy), Group D ( combined ionizing radiation, treated with ionizing radiation from low dose to high dose : 50cGy first and 300cGy after 6h interval). The mRNA levels of HIF-1 and VEGF in each group were detected by real time polymerase chain reaction, while HPSE-1 expression was measured by ELISA. The microvessel density (MVD) and hypoxic cells were determined through immunohistochemical (IHC) staining of CD31 and HIF-1a. Results: Significant differences of HIF-1${\alpha}$ mRNA level could be found among the 4 groups (F=74.164, P<0.001): Group C>Group A>Group D> Group B. The mRNA level of VEGF in Group C was significantly higher than in the other three groups (t=-5.267, P=0.000), while no significant difference was observed among Group A, B and D (t=1.528, 1.588; P=0.205, 0.222). In addition, the MVD was shown to be the highest in Group C (t=6.253, P=0.000), whereas the HPSE-1 level in Group A was lower than in Group B (t=14.066, P=0.000) and higher than in Group C (t=-21.919, P=0.000), and similar with Group D (t=-2.066, P=0.058). Through IHC staining of HIF-1a, the expression of hypoxic cells in Group A was (++), Group B was (+), Group C was (+++) and Group D was (+). Conclusion: Ionizing radiation with lowerdoses might improve tumor hypoxia through inhibiting the expression of HIF-1 and HPSE-1, whereas higherdoses worsen tumor hypoxic conditions by up-regulating HIF-1${\alpha}$, HPSE-1, VEGF and CD31 levels. A protocol of low-dose ionizing radiation followed by a high-dose irradiation might at least partly improve tumor hypoxia and enhance radiosensitivity.

Bevacizumab accelerates corneal wound healing by inhibiting TGF-βexpression in alkali-burned mouse cornea

  • Lee, Sung-Ho;Leem, Hyun-Sung;Jeong, Seon-Mi;Lee, Koon-ja
    • BMB Reports
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    • v.42 no.12
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    • pp.800-805
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    • 2009
  • This study investigated the effect of subconjunctival injections of bevacizumab, an anti-VEGF antibody, on processes involved in corneal wound healing after alkali burn injury. Mice were divided into three groups: Group 1 was the saline-treated control, group 2 received subconjunctival injection of bevacizumab 1hr after injury and group 3 received bevacizumab 1 hr and 4 days after injury. Cornea neovascularization and opacity were observed using a slit lamp microscope. Corneal repair was assessed through histological analysis and immunostaining for CD31, $\alpha$-SMA, collagen I, and TGF-$\beta$2 7 days post-injury. In group 3, injection of bevacizumab significantly lowered neovascularization and improved corneal transparency. Immunostaining analysis demonstrated a reduction in CD31, $\alpha$-SMA and TGF-$\beta$2 levels in stroma compared to group 1. These results indicate that bevacizumab may be useful in reducing neovascularization and improving corneal transparency following corneal alkali burn injury by accelerating regeneration of the basement membrane.

Morphologic Evidence of Anti-Tumor Specificity of T Cells Activated by Denritic Cells Derived from Peripheral Blood Mononuclear Cells of Thyroid Cancer Patients

  • Lee, Dae-Heui
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.4
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    • pp.243-247
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    • 2012
  • Recent studies suggest that immunization with autologous dendritic cells (DCs) results in protective immunity and rejection of established tumors in various human malignancies. The purpose of this study is to determine whether DCs are generated from peripheral blood mononuclear cells (PBMNs) by using cytokines such as F1t-3 ligand (FL), granulocyte macrophage-colony stimulating factor (GM-CSF), IL-4, and TNF-${\alpha}$, and whether cytotoxic T cells activated against the thyroid cancer tissues by the DCs. Peripheral blood was obtained from 2 patients with thyroid cancer. DCs were established from PBMNs by culturing in the presence of FL, GM-CSF, IL-4, and TNF-${\alpha}$ for 14 days. At day 14, the differentiated DCs was analyzed morphologically. The immunophenotypic features of DCs such as CDla, CD83, and CD86 were analyzed by immunofluorelescence microscopy. At day 18, DCs and T cells were incubated with thyroid cancer tissues or normal thyroid tissues for additional 4 days, respectively. DCs generated from the PBMNs showed the typical morphology of DCs. Activated cytotoxic T lymphocytes (CTLs) were observed also. DCs and the CTLs were attached to the cancer tissues on scanning electron microscope. The DCs activated the CTLs, which able to specifically attack the thyroid cancer. This study provides morphologic evidence that the coculture of T cells/cancer tissues activated the T cells and differentiated CTLs. The CTLs tightly adhered to cancer tissues and lysed cancer tissues vigorously. Therefore DCs could be used as potential vaccines in the immunotherapy.

Reaarding Effect of Dietary Fibers Isolated from Tangerine Peels on Glucose, Bile Acid, Cadmium transport In Vitro (감귤과피로부터 분리한 식이섬유의 포도당, 담즙산, 카드뮴 투과억제에 관한 In Vitro 연구)

  • 김미경
    • Journal of Nutrition and Health
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    • v.30 no.2
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    • pp.210-219
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    • 1997
  • Retarding effects of the dietary fibers from tangerine peels on glucose, bile acid and cadmium transport were evaluated by dialysis method, and were compared with those of commercial dietary fibers(citrus pection, CM-cellulose, guar gum, $\alpha$-cellulose). Yields of total (TDF), insoluble(IDF) and soluble dietary fibers(SDF) from tangerine peels on the fresh matter basis were 2.84%, 1.95% and 0.39% respectively. The amount of insoluble fibers was 5.2 times higher than that of soluble fibers. Soluble fibers(guar gum, CM-cellulose, SDF, pectin) had the retarding effect on glucose transport, while IDF, TDF and $\alpha$-cellulose did not have. Guar gum showed the greatest effect, followed by CM-cellulose, SDF and pectin. Among the extracted fibers, only SDF had the effect on glucose transport retardation. Regarding bile acid dialysis, guar gum had the greatest retarding effect, and all dietary fibers from tangerine peels, especially SDF, showed the effect of bile acid retardation. On cadmium transport retardation, CM-cellulose had the greatest effect, followed by SDF, TDF, IDF, guar gum and pectin. Among the extracted fibers, SDF had the greatest effect on Cd trasport transport retardation. The extracted dietary fibers showed higher retarding effect on Cd transport than glucose and bile acid transport, and the effect of SDF was higher than IDF.

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Development of a Cardiovascular Disease Resilience Scale (심혈관질환용 회복력(Cardiovascular Disease Resilience) 측정도구 개발 및 평가)

  • Shin, Su-Jin
    • Korean Journal of Adult Nursing
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    • v.22 no.2
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    • pp.161-170
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    • 2010
  • Purpose: The purpose of this study was to develop a Cardiovascular Disease Resilience (CDR) scale to evaluate disease specific resilience for recovery. Methods: The study was conducted as follows: items generation, and test of validity and reliability. Items were developed via literature review, review of instruments, and data acquired from the interviews. In order to test validity and reliability, seven panels of experts reviewed the preliminary questionnaire and then data were collected from 550 cardiovascular disease patients. Factor analysis, Pearson correlation, ANOVA, and Cronbach's alpha were used to analyze the data. Results: In the preliminary stage, forty-four items were generated. A reduction to 40 items was accomplished through content validity analysis. Factor analysis extracted 7 factors with a total of 25 items. The CDR items were moderately correlated with the subscales of the CD-RISC (Connor-Davidson Resilience Scale) and the mean score of CDR was associated with quality of life measured with CD-QOL (Cardiovascular Disease Quality of Life). Cronbach's ${\alpha}$=.84. Conclusion: Content validity, construct validity, criterion validity, and reliability of the CDR were established. The CDR is a reliable and valid instrument which the resilience of cardiovascular disease specific recovery state can be evaluated.