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Effects of fermented Taraxacum mongolicum by Dongchimi on protection of liver cell and immune activity (포공영(민들레)의 동치미 발효에 의한 간세포 보호 및 면역증진 효과)

  • Choi, Bog-Soo;Kim, Hye-Ja
    • Journal of Society of Preventive Korean Medicine
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    • v.18 no.1
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    • pp.11-21
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    • 2014
  • Objective : This study set out to combine the treatment efficacy of Taraxacum with Dongchimi fermentation and investigate Taraxacum's effects on protection of liver cell and controlling nitric oxide(NO) through experiments, thus checking whether it had values as a physiological active matter. The experimental materials include Taraxacum Dongchimi (TD) and Taraxacum fermented by Dongchimi (TDF). As for methodology, experiments were carried out to compare TD and TDF in components, protection effects for liver cells, anticancer effects on liver cells, and protection effects for brain cells in the aspects of liver function and immunity enhancement. Method : The experimental materials include Taraxacum Dongchimi (TD) and Taraxacum fermented by Dongchimi (TDF). As for methodology, experiments were carried out to compare TD and TDF in components, protection effects for liver cells, anti-cancer effects on liver cells, and protection effects for brain cells in the aspects of liver function and immunity enhancement. Results : As shown in the chromatogram results, each valid component content increased in Taraxacum fermented by Dongchimi (TDF) for each time section. Of them, the valid component content at 36.80 minutes was approximately 2.7 times higher in TDF at 21.8% than in Taraxacum Dongchimi (TD) at 8.28%. TDF generated more excellent protection effects against the toxicity that caused oxidative damage to the liver cell(HepG2) with t-BHP than TD. The survival rate was low in TD of $100{\mu}g/m{\ell}$ and $300{\mu}g/m{\ell}$ and increased to 23.3% in TDF of $100{\mu}g/m{\ell}$. The survival rate was the highest at $300{\mu}M$ with a significant difference of 68.1%(P<0.05). Both TD and TDF showed effects of controlling nitric oxide production according to concentration with TDF recording a higher rate of controlling nitric oxide production than TD. There were significant differences(P<0.05) in the effects of controlling nitric oxide production at 200 ug/ml, 400 ug/ml in both groups. Especially the result TDF of $400{\mu}g/m{\ell}$ was thus similar to those of butein, the positive control group. Conclusion : The result of this studies is that Taraxacum fermented by Dongchimi (TDF) increased the valid component content compared with the simple mixture(TD). The findings clearly show that it is a material with the effects of improving immunity and liver cell protection. If fermentation methods are further developed to use it as a functional material, it will be subject to more opportunities of being used in other functional foods and make a contribution to integrated medicinal food development.

Properties of Extremely Low Frequency Electromagnetic Fields and their Effects on Mouse Testicular Germ Cells

  • Kim, Yeon-Sook;Lee, Suk-Keun
    • International Journal of Oral Biology
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    • v.35 no.4
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    • pp.137-144
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    • 2010
  • To evaluate the biohazard properties of an extremely low frequency electromagnetic field (ELF-EMF), we explored the physical properties of the ELF-EMF that generates the electric current induction in the secondary coil from the chamber of a primary solenoid coil. We subsequently explored the biological effects of a strong alternating electromagnetic field (EMF), ranging from 730-960 Gauss, on the mouse testis. Mice were exposed to an alternating EMF field induced by a rectangular electric current at 1, 7, 20, 40, and 80 Hertz, for 1, 3, 5, and 7 hours. The mouse testes were examined for proliferative activity and apoptosis using the in situ terminal deoxynucleotidyl transferase (TdT) method and by immunostaining of proliferating cell nuclear antigen (PCNA), respectively. We found that the electric currentm induction increased in the 6-8 Hertz range, and that exposure to an ELF-EMF induced the apoptosis of mouse spermatocytes. In situ TdT staining was found to be most prominent in 7 Hertz group, and gradually reduced in the 20, 40, and 80 Hertz groups. These data suggest that a strong EMF can induce reproductive cell death within a short time, and the harmful effects of the EMF are maximal at low frequency alternating EMFs.

A PRELIMINARY STUDY OF B LYMPHOCYTE ANTIGEN D8/17 IN TOURETTE SYNDROME CHILDREN WHO SUGGESTED PANDAS (PANDAS가 의심되는 뚜렛장애 아동에서의 B 임파항원 D8/17에 관한 예비연구)

  • Joung Yoo-Sook;Lee Young-Sik
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.17 no.1
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    • pp.27-31
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    • 2006
  • Objectives : We examined whether D8/17 expression in Tourette syndrome children who suggested PANDAS were higher than comparison group, and there was my clinical difference by D8/l7 expression. Methods : Nine Tourette's syndrome children suggested PANDAS and two ADHD children without tic disorder were evaluated far percentage of D8/17 expression-positive B cells by immunofluorescence flow cytometric assay and anti-streptolysin O titer. Results : The frequency of D8/17 positive B lymphocyte rate was significantly higher in Tourette's syndrome than ADHD, whose average rate were 77.9 and 24.8, respectively. Among 9 TD patients,4 patients showed above 90% D8/l7 expression. There was high concordance expression rate between mother (98.4%) and daughter (99.0%) The significant relation between percentage of D8/17 expression and tic severity were not detected. The significant relation between percentage of D8/17 expression and anti-streptolysin O titer were not detected, however in 66.7% TD patients showed above 100IU/ml. Conclusion : We concluded that subgroup of TD children are streptococcal infected tic disorder, so called PANDAS.

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Effect of growth hormone on migration and tumover rats of small intestinal epithelium in mice (성장(成長) hormone이 랫드 소장상피세포(小腸上皮細胞) 재생주기(再生週期)에 미치는 영향에 대하여)

  • Kwak, Soo-dong;Kim, Chong-sup;Ahn, Dong-weon
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.37-42
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    • 1993
  • The purpose of this study was to examine the effect of growth hormone(GH) on the division and migration of the small intestinal epithelium in mice. Twenty mice(ICR), weighing initially about 10 g, aged 18 days were randomly subdivided in two groups of control and GH-treated group. Control group was sacrificied at 2 hr, 2, 3, 4 and 5 day after intraperitoneal injection of $^3H$-thymidine($^3H$-TdR, $4{\mu}$ Ci/gm of BW/day for 2 days). GH-treated group was injected subcutaneously with somatotropin(10 IU/mouse/day for 4 days) from 2 day ago of first $^3H$-TdR injection and then was sacrificied at 2 hr, 2, 3, 4 and 5 day after intraperitoneal injection of $^3H$-TdR($4{\mu}$ Ci/g of BW/day for 2 days). The small intestines were collected for autoradiogrophy and Hematoxylin counterstain. The location of the labeled epithelial cells(LEC) in villi of the small intestine was invested with light microscope. 1. In the control group, the LEC regions in the small intestine were located at crypts on 1 dsy, at $0%{\sim}15.9{\pm}3.6%$ of villus high value(VHV) on 2 day, at $0%{\sim}49.8{\pm}16.5%$ of VHV on 3 day, at $0%{\sim}95.3{\pm}6.9%$ of VHV on 4 day and $62.9{\pm}16.7{\sim}100%$ of VHV on 5 day, respectively. 2. In the GH-treated group, the LEC regions in the small intestine were located at crypts on 1 day, at $0%{\sim}39.2{\pm}9.5%$ of VHV on 2 day, at $0%{\sim}81.5{\pm}18.2%$ of VHV on 3 day, at $45.2{\pm}11.5%{\sim}100%$ of VHV on 4 day, at 85%~100% of VHV on 5 day, respectively. 3. VHV of tops in the LEC regions appeared to be 23.3% and 31.7% higher on 2 and 3 day, and VHV of the LEC region bases appeared to be 45.2%, 22.1% higher on 4 and 5 day, respectively in the GH-treated group than those observed in the control group. These difference was very highly significant(p<0.01).

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Intracellular Mechanisms of Growth Hormone Action on Apoptosis in Cultured Porcine Ovarian Granulosa Cells

  • Sirotkin, A.V.;Makarevich, A.V.;Pivko, J.;Genieser, H.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1045-1050
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    • 2002
  • The aims of this study were to detect spontaneously occurring apoptosis in cultured porcine ovarian cells, to examine the role of growth hormone (GH), tyrosine kinase (TK), protein kinase G (PKG) and cyclin-dependent kinase (CDK) in the control of this process, and to determine whether the effect of GH on apoptosis is mediated by TK-, PKG- and cdc2-dependent intracellular mechanisms. We studied the action of pGH (10 ng/ml), blockers of TK (genistein, lavendustin, both 100 ng/ml), PKG (Rp-Br-PET-cGMPS, 50 nM; KT5823, 100 ng/ml) and CDK (olomoucine, $1{\mu}g/ml$), as well as combinations of GH with these blockers, on the onset of apoptosis in cultured granulosa cells isolated from antral (3-6 mm) porcine follicles. The functional characteristics of an early apoptotic event, DNA fragmentation, were determined using terminal deoxynucleotidyltransferase (TdT)-mediated dUTP nick end labelling (TUNEL), whilst morphological signs of advanced apoptosis such as pyknosis, chromatin marginalization, shrinkage and fragmentation of nucleus, were detected using routine light microscopy. After culture, some ovarian granulosa cells exhibited DNA fragmentation, which in some cases was associated with morphological apoptosis-related changes (pyknosis, shrinkage and fragmentation of the nucleus). GH significantly reduced the proportion of TUNEL-positive cells. Neither TK nor CDK blockers when given alone, significantly affected the percentage of TUNEL-positive cells although both PKG blockers significantly increased this index. Furthermore, TK and PKG blockers given together with GH, prevented or reversed the inhibitory effect of GH on apoptosis, whilst the CDK blocker olomoucine promoted it. These observations demonstrate apoptosis in porcine ovaries and suggest the involvement of GH, TK, PKG and CDK in the control of this process. They also suggest that the effect of GH on ovarian apoptosis is mediated or regulated by multiple signalling pathways including TK-, PKG- and CDK-dependent intracellular mechanisms.

Pharmacodynamics of CKD-602 (Belotecan) in 3D Cultures of Human Colorectal Carcinoma Cells

  • Lee Sin-Hyung;Al-Abd Ahmed M.;Park Jong-Kook;Cha Jung-Ho;Ahn Soon-Kil;Kim Joon-Kyum;Kuh Hyo-Jeong
    • Biomolecules & Therapeutics
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    • v.14 no.2
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    • pp.90-95
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    • 2006
  • CKD-602 exerts its antitumor effect via inhibition of topoisomerase I in cancer cells. Multicellular spheroid (MCS) and Multicellular layers (MCLs) are known as in vitro 3-dimensional models which closely represent tumor conditions in vivo. In order to investigate the potential of CKD-602 against human colorectal tumors, we evaluated the anti-proliferative activity and penetration ability of CKD-602 in MCS and MCL cultures of DLD-l human colorectal cancer cells, respectively. The maximum effects($E_{max}$) induced by CKD-602 were significantly lower in MCS compared to monolayers (48% vs 92%). With prolonged drug exposure, the $IC_{50's}$ of CKD-602 decreased to $23.5{\pm}1.0nM$ in monolayers after 24 h exposure and $42.3{\pm}1.7nM$ in MCS after 6 days, respectively. However, no further increase in effect was observed for exposure time longer than growth doubling time (Td) in both cultures. Activity of CKD-602 was significantly reduced after penetration through MCL and also with cell-free insert membrane. In conclusion, CKD-602 showed significantly decreased anti-proliferative activity in 3D cultures (MCS) of human colorectal cancer cells. Tumor penetration of CKD-602 could not be determined due to loss of activity after penetration through cell free insert membrane, which warrants further evaluation using a modified model.

Surface Marker Analysis in Acute Leukemias (백혈병의 면역학적 표지자검사의 결과분석)

  • Moon, Jin-Young;Lee, Chae-Hoon;Kim, Kyung-Dong;Kim, Chung-Sook
    • Journal of Yeungnam Medical Science
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    • v.14 no.2
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    • pp.359-369
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    • 1997
  • We studied the expression of the cell surface antigen associated with myeloid and lymphoid leukemias on bone marrow or peripheral blood blast cells from 153 leukemic patients including 61 cases of acute myelogenous leukemias(AML), 46 of acute lymphocytic leukemias(ALL) and 12 of acute leukemias. They were analyzed by direct or indirect immunofluorescence method for reactivity with the monoclonal antibodies to B cells(CD10, CD19, SmIg), T cells(CD2, CD5, CD7, CD3, CD4, CD8), myeloid antigen(CD13, CD14, CD33, CD61) and a nonspecific antigen, HLA-DR. Lymphoid associated markers detected on AML is CD7 32.8%, CD10 14.8%, CD5 13.1%, CD2 6.6% and CD19 1.6%. TdT was positive in 4.9% of AMLs. Hybrid leukemias were 8 cases out 61 AML cases and were mainly composed of monocytic lineage, M4 and M5a. Myeloid markers detected in ALL were CD13 2.2% and CD33 2.2%. In this study, immunologically classified ALLs were composed of 65.2% of CALLA (+) B precursor type, 10.9% of CALLA (-) B precursor pattern, 8.7% of T cell type, 2.2% of B cell type, 4.5% of mixed lymphoid lineage(B&T), 2.2% of undifferentiated leukemia, and 6.5% of hybrid leukemia. Twelve cases of acute leukemias ware finally diagnosed to be 5 cases of hybrid leukemia, 3 cases of B lineage, 3 case of T lineage and 1 case of mixed lymphoid(B&T) leukemia. In summary, we think the best method for typing acute leukemias is by using a combination of FAB classification and immunophenotying.

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When Dose Losses of Maternal Lymphocytes Response to Trophoblast Antigen or Alloantigen Occur in Women with a History of Recurrent Spontaneous Abortion? (반복유산을 경험한 환자에서 임신중 태반항원과 동종항원에 노출된 모체 림프구면역반응은 언제부터 소실되나?)

  • Choi, Bum-Chae;Hill, Joseph A.
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.115-122
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    • 1998
  • The maintenance of a viable pregnancy has long been viewed as an immunological paradox. The deveolping embryo and trophoblast are immunologically foreign to the maternal immune system due to their maternally inherited genes products and tissue-specific differentiation antigens (Hill & Anderson, 1988). Therefore, speculation has arisen that spontaneous abortion may be caused by impaired maternal immune tolerance to the semiallogenic conceptus (Hill, 1990). Loss of recall antigen has been reported in immunosuppressed transplant recipients and is associated with graft survival (Muluk et al., 1991; Schulik et al., 1994). Progesterone $(10^{-5}M)$ has immunosuppressive capabilities (Szekeres-Bartho et al., 1985). Previous study showed that fertile women, but not women with unexplained recurrent abortion (URA), lose their immune response to recall antigens when pregnant (Bermas & Hill, 1997). Therefore, we hypothesized that immunosuppressive doses of progesterone may affect proliferative response of lymphocytes to trophoblast antigen and alloantigen. Proliferative responses using $^3H$-thymidine ($^3H$-TdR) incorporation of peripheral blood mononuclear cells (PBMCs) to the irradiated allogeneic periperal blood mononuclear cells as alloantigen, trophoblast extract and Flu as recall antigen, and PHA as mitogen were serially checked in 9 women who had experienced unexplained recurrent miscarriage. Progesterone vaginal suppositories (100mg b.i.d; Utrogestan, Organon) beginning 3 days after ovulation were given to 9 women with unexplained RSA who had prior evidence of Th1 immunity to trophoblast. We checked proliferation responses to conception cycle before and after progesterone supplementation once a week through the first 7 weeks of pregnancy. All patients of alloantigen and PHA had a positive proliferation response that occmed in the baseline phase. But 4 out of 9 patients (44.4%) of trophoblast antigen and Flu antigen had a positive proliferative response. The suppression of proliferation response to each antigen were started after proliferative phase and during pregnancy cycles. Our data demonstrated that since in vivo progesterone treated PBMCs suppressed more T-lymphocyte activation and $^3H$-TdR incorporation compare to PBMCs, which are not influenced by progesterone. This data suggested that it might be influenced by immunosuppressive effect of progesterone. In conclusion, progesterone may play an important immunological role in regulating local immune response in the fetal-placental unit. Furthermore, in the 9 women given progesterone during a conception cycle, Only two (22%) repeat pregnancy losses occured in these 9 women despite loss of antigen responsiveness (one chemical pregnancy loss and one loss at 8 weeks of growth which was karyotyped as a Trisomy 4). These finding suggested that pregnancy loss due to fetal aneuploidy is not associated with immunological phenomena.

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Activation of Pro-Apoptotic Multidomain Bcl-2 Family Member Bak and Mitochondria-Dependent Caspase Cascade are Involved in p-Coumaric Acid-Induced Apoptosis in Human Jurkat T Cells (p-Coumaric acid에 의해 유도되는 인체 Jurkat T 세포의 에폽토시스 기전)

  • Lee, Je-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1678-1688
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    • 2011
  • The apoptogenic effect of p-coumaric acid, a phenolic acid found in various edible plants, on human acute leukemia Jurkat T cells was investigated. Exposure of Jurkat T cells to p-coumaric acid (50-$150{\mu}M$) caused cytotoxicity and TdT-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic DNA fragmentation along with Bak activation, ${\Delta}{\psi}m$ loss, activation of caspase-9, -3, -7, and -8, and PARP degradation in a dose-dependent manner. However,these apoptotic events were completely abrogated in Jurkat T cells overexpressing Bcl-2.Under these conditions, necrosis was not accompanied. Pretreatment of the cells with the pan-caspase inhibitor (z-VAD-fmk) could prevent p-coumaric acid-induced sub-$G_1$ peak representing apoptotic cells, whereas it failed to block ${\Delta}{\psi}m$ loss, indicating that the activation of caspase cascade was prerequisite for p-coumaric acid-induced apoptosis as a downstream event of ${\Delta}{\psi}m$ loss. FADD- and caspase-8-positive wild-type Jurkat T cell clone A3, FADD-deficient Jurkat T cell clone I2.1, and caspase-8-deficient Jurkat T cell clone I9.2 exhibited similar susceptibilities to the cytotoxicity of p-coumaric acid, excluding an involvement of Fas/FasL system in triggering the apoptosis. The apoptogenic activity of p-coumaric acid is more potent in malignant Jurkat T cells than in normal human peripheral T cells. Together, these results demonstrated that p-coumaric acid-induced apoptogenic activity in Jurkat T cellswas mediated by Bak activation, ${\Delta}{\psi}m$ loss, and subsequent activation of multiple caspases such as caspase-9, -3, -7, and-8, and PARP degradation, which could be regulated by anti-apoptotic protein Bcl-2.