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Condurango (Gonolobus condurango) Extract Activates Fas Receptor and Depolarizes Mitochondrial Membrane Potential to Induce ROS-dependent Apoptosis in Cancer Cells in vitro -CE-treatment on HeLa: a ROS-dependent mechanism-

  • Bishayee, Kausik;Mondal, Jesmin;Sikdar, Sourav;Khuda-Bukhsh, Anisur Rahman
    • Journal of Pharmacopuncture
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    • v.18 no.3
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    • pp.32-41
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    • 2015
  • Objectives: Condurango (Gonolobus condurango) extract is used by complementary and alternative medicine (CAM) practitioners as a traditional medicine, including homeopathy, mainly for the treatment of syphilis. Condurango bark extract is also known to reduce tumor volume, but the underlying molecular mechanisms still remain unclear. Methods: Using a cervical cancer cell line (HeLa) as our model, the molecular events behind condurango extract's (CE's) anticancer effect were investigated by using flow cytometry, immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Other included cell types were prostate cancer cells (PC3), transformed liver cells (WRL-68), and peripheral blood mononuclear cells (PBMCs). Results: Condurango extract (CE) was found to be cytotoxic against target cells, and this was significantly deactivated in the presence of N-acetyl cysteine (NAC), a scavenger of reactive oxygen species (ROS), suggesting that its action could be mediated through ROS generation. CE caused an increase in the HeLa cell population containing deoxyribonucleic acid (DNA) damage at the G zero/Growth 1 (G0/G1) stage. Further, CE increased the tumor necrosis factor alpha ($TNF-{\alpha}$) and the fas receptor (FasR) levels both at the ribonucleic acid (RNA) and the protein levels, indicating that CE might have a cytotoxic mechanism of action. CE also triggered a sharp decrease in the expression of nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) both at the RNA and the protein levels, a possible route to attenuation of B-cell lymphoma 2 (Bcl-2), and caused an opening of the mitochondrial membrane's permeability transition (MPT) pores, thus enhancing caspase activities. Conclusion: Overall, our results suggest possible pathways for CE mediated cytotoxicity in model cancer cells.

Effect of Lithospermi Radix on Contact Dermatitis Induced by Dinitrofluorobenzene in Mice

  • Kim, Han-Na;Kim, Mi-Young;Choi, Chan-Hun;Kim, Byung-Joo;Kim, Kyung-Yoon;Kim, Gye-Yeop;Jeong, Hyun-Woo;Kim, Hyung-Woo
    • Journal of Pharmacopuncture
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    • v.15 no.2
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    • pp.7-10
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    • 2012
  • Objectives: The root of Lithospermum erythrorhizon Sieb. et Zucc. (Lithospermi Radix, LR) is a kind of heat clearing and blood cooling medicinal herbs. It can clear away heat and cool the blood, reduce toxins and disperse maculae. LR has long been used as efficacious therapy for inflammation, burns, frostbite and skin diseases such as eczema and psoriasis. Methods: In the present study, we investigate anti-allergic and anti-inflammatory effects of LR by using the 1-fluoro-2, 4- dinitrofluorobenzene (DNFB)-induced contact dermatitis mouse model. Results: Topical application of 10 mg/mL of LR effectively inhibited skin lesions induced by repeated paintings with DNFB. Topical application of LR also inhibited hyperplasia, edema, spongiosis and infiltrations of mononuclear cells. In addition, production levels of total immunoglobulin and IgG1 in serum were decreased by using LR in vivo. Conclusions: These data suggest that LR acts as an antiinflammatory agent, improving skin lesions in CD mice.

Expression of Heat Shock Protein in Cytokine Stimulated PDL Cells and Inflamed Gingival Tissue (염증성 치은조직과 치주인대세포에서 Cytokine에 의해 유도되는 열충격단백 발현에 관한 연구)

  • Cho, In-Ho;Kim, Doek-Kyu;Kim, Eun-Cheol;You, Hyung-Keun;Shink, Hyung-Shin
    • Journal of Periodontal and Implant Science
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    • v.28 no.1
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    • pp.103-120
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    • 1998
  • Prokaryotic and eukaryotic cells respond to heat stress and other environmental abuses by synthesizing a small set of stress proteins and by inhibiting post-transcription synthesis of normal proteins. The purpose of the present study was to document the stress response produced by inflamed gingival tissue in vivo, and cytokine inducted human periodontal ligament cells. Human PDL cells were exposed to TNF-$\alpha$(1ng/ml), INF-$\gamma$(200 U/ml), LPS(100ug/ml), combination of cytokine, and SDS-PAGE gels running and Western blotting analysis was done. In vivo studies, the healthy gingival tissusse of a control group and inflamed gingival tissue of adult periodontitis were studied by immunohistochemistry and histology. The results were as follows 1. HSP 47 was distributed on basal layer in healthy gingiva, but stronger stained in basal, suprabasal, and spinous layer of inflamed gingiva. 2. HSP 47 was rare on endothelial cells and mononuclear cells in healthy gingiva, but stronger expressed in inflamed gingiva. 3. HSP 70 expression was rare on epihelium and inflammatory cells hi both healthy & inflamed gingiva. 4. HSP 70 was actively expressed on endothelial cells and inflammatory cells of capillary lumen in moderately & mild inflamend gingiva. 5. PDL cells showed low level of HSP 47 protein expression which was significantly induced by cytokine stimulation (LSP only and combination). 6. Maximum HSP 70 protein induction was seen with stimulation by a combination of the cytokine, Combination of TNF-$\alpha$, INF-$\gamma$, LPS have been shown to synergistically effects of HSP 70 expression. On the above findings, HSP Is influenced by cytokine and chronic inflammation in vivo, and may be involved in protection of tissue during periodontal inflammatiom.

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Anticancer Effect of Activated Natural Killer Cells on Human Colorectal Tumor (결장암에 대한 활성 자연살해세포의 항암효능)

  • Sung, Hye-Ran;Kim, Jee-Youn;Park, Min-Gyeong;Kim, Il-Hoi;Lee, Dong-Wook;Han, Sang-Bae;Lee, Chong-Kil;Song, Suk-Gil
    • YAKHAK HOEJI
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    • v.54 no.3
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    • pp.192-199
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    • 2010
  • Colorectal cancer is one of the most common alimentary malignancies. In this study, the antitumor activity of activated human natural killer (NK) cells against human colorectal cancer was evaluated in vivo. Human NK cells are the key contributors of innate immune response and the effective functions of these cells are enhanced by cytokines. Human peripheral blood mononuclear cells (PBMC) were cultured with interleukin-2 (IL-2)-containing medium for 14 days and resulted in enriched NK cell population. The resulting populations of the cells comprised 7% $CD3^+CD4^+$ cells, 25% $CD3^+CD8^+$ cells, 13% $CD3^-CD8^+$ cells, 4% $CD3^+$CD16/$CD56^+$ cells, 39% $CD3^+$CD16/$CD56^-$ cells, and 52% $CD3^-$CD16/$CD56^+$ cells. Tumor necrosis factor alpha (TNF-$\alpha$), interferon gamma (IFN-$\gamma$), IL-2, IL-4, and IL-5 transcripts of the activated NK cells were confirmed by RT-PCR. In addition, activated NK cells at doses of 2.5, 5 and 10 million cells per mouse inhibited 10%, 34% and 47% of SW620-induced tumor growth in nude mouse xenograft assays, respectively. This study suggests that NK cell-based immunotherapy may be used as an adoptive immunotherapy for colorectal cancer patients.

Effect of 1,3-Benzopyrone on Chemotactic Activity of Peripheral Blood Leukocytes in the Dog (1,2-benzopyrone이 개 말초혈액 백혈구의 유주활성에 미치는 영향)

  • Kim Jee-hyun;Kang Ji-houn;Yang Mhan-pyo
    • Journal of Veterinary Clinics
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    • v.22 no.2
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    • pp.84-89
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    • 2005
  • This study was undertaken to examine whether 1,2-benzopyrone affects on chemotactic activity of canine peripheral blood leukocytes. A modified Boyden chamber method was sed on chemotaxis evaluation. The direct treatments of 1,2-benzopyrone showed no ffects on the chemotaxis of peripheral blood mononuclear cells (PBMCs) and olymorphonuclear cells (PMNs). But chemotaxis of PMN was remarkably enhanced by ulture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similarly, it as also increased by recombinant (r) interleukin (lL)­8. This chemotactic activity of MN was inhibited by addition of anti-rIL-8 polyclonal antibody. The chemotaxis of PBMC was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. Therefore, these results suggested that the chemotactic activity of PMN ay be mainly mediated by IL-8-like factor(s) produced from PBMC treated with ,2-benzopyrone.

Effects of Herba, Radix, Fructus-xanthii Extract on the Immunities against infections Diseases and Tumors (창이초(蒼耳草)의 약용부위별(藥用部位別) 추출물이 항감염(抗感炎) 및 항종양(抗腫瘍) 면역반응(免疫反應)에 미치는 영향)

  • Cho, Nam-Zoon;Song, Ho-Joon;Shin, Min-Kyo
    • The Journal of Korean Medicine
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    • v.19 no.2
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    • pp.420-438
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    • 1998
  • Herba Xanthii(HX), Radix Xantluii(RX) and Fructus Xanthii(FX) is one of the oriental medicine that has been used for the treatment of such infectious diseases and tumors. However, the mechanism of the drug is not investigated much. This study was done to know the effects of HX, RX and FX extract on the such innate immunities as phagocytic function and reactive radical formtions from phagocytes and the such acquired immunities as humoral and cell-mediated immunities. The followings are the results obtained from this study: 1. HX2 and FX1 groups increases the in vivo phagocytic activity of mononuclear phagocytes. 2. HXB, RXB, RXC, FXB and FXC groups increase the in vitro phagocytic activities. 3. RXB group stimulated the macrophages to produce nitric oxide in the presence of $interferon-{\gamma}$ $(IFN-{\gamma})$. 4. HX and RX whole groups increased the luminol-amplified reactive oxygen intermediate production in vivo. 5. HX whole and RX1, FX2 groups increased the lucigenin-amplified reactive oxygen intennediate production in vivo. 6. HXC group only increased the luminol-amplified reactive oxygen intermediate production in vitro. 7. HXB, FXB and FXC groups increased the lucigenin-amplified reactive oxygen intermediate production in vitro. 8. HX2, RX1 and FX whole groups increased the hemolysin formations from B cells. 9. HX, RX and FX whole groups significantly increased the rosette forming cells from the spleen. 10. HX, RX and FX whole groups significantly decreased the delayed-type hypersensitivity measured by footpad swelling. The above results demonstrate that HX, RX and FX has enhancing effects on innate immunity selectively and decreasing effects on delayed-type hypersensitivity of cell-mediated immunity according to medicinal part and diluted condition. This immunomodulating effects of HX, RX and FX might be responsible for the treatment of immune-mediated disorders.

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Effect of Edible and Medicinal Plants on the Activation of Immune Cells (생약제가 면역세포 활성화에 미치는 영향)

  • 이인선;하영득
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.1
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    • pp.150-155
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    • 1994
  • In order to evaluate the effect of the extracts of eidble and medicinalplant son the activation of immune cells, measurements were made by ELISA and radioimmunoassay on the degree of release for the tumor necrosis factor (TNF) and neopterin by the edible and medicinal plants in peripheral blood cells. The results of measurements of TNF in the supernatant cultured liquid showed nothing in t도 control which does not have any edible and medicinal plants. However, measurements of TNF 9pg/ml) in the samples are given as follows : 716.7 in lipopolysaccharide (LPS 1 g/ml), 465.2 Rheum plamatum L.m302.7 Sanguisorba offciinalis L. 818.2 Rubus coreanus M, 328.3 Terminalia chebula R., 426.6 AReca catechu L. 227.0 Eugenia caryophiliata T., 272.9 Ephedra sinica S., 30.1 Caesalpinia sappan L., 474.0 Chaenomeles japonica L., 396.0 Cornus officinalis S.in edible and medicinalplants. ENopterin (n mole/L) value showed below the check point in the control group, however, the values are 11.0 in LPS, and edible and medicinal plants, 5.3 Rheum palmatum L., 11.6 Eugenia caryophiliata T., 5.5 Ephedra sinica S., 4.5 Caesalpinia sappan L., 4.3 Chaenomelees japonica L.3.7 Cornus officinalis S. In order to find m RNA levels of Cytokines increased by edible and medicinal plants, total RNA was separated from mononuclear cells treated 5 hrs with Rubus coreanus M. and then administrated for RT-PCR. The considerable increases of the m RNA of TNF, IL-1 $\alpha$ and IL-6 were observed.

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Korean Red Ginseng Improves Vascular Stiffness in Patients with Coronary Artery Disease

  • Chung, Ick-Mo;Lim, Joo-Weon;Pyun, Wook-Bum;Kim, Hye-Young
    • Journal of Ginseng Research
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    • v.34 no.3
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    • pp.212-218
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    • 2010
  • Korean red ginseng (KRG) has been shown to enhance endothelium-dependent vasorelaxation in experimental animals; however, little is known about its pharmacological effects on vascular stiffness in patients with coronary artery disease (CAD). This randomized, double-blind, placebo-controlled crossover trial was carried out to determine whether KRG has beneficial effects on arterial stiffness, cardiovascular risk factors such as plasma lipid profiles and blood pressure (BP), and Rho-associated kinase (ROCK) activity. Twenty patients (mean age, 62.5 years) with stable angina pectoris were given KRG (2.7 g/day) and a placebo alternatively for 10 weeks. Blood biochemical analysis and pulse wave velocity (PWV) recording were performed on day 0 and after the completion of each treatment. ROCK activity was assessed based on the level of phospho-$Thr^{853}$ in the myosin-binding subunit of myosin light chain phosphatase, determined by Western blot analysis of peripheral blood mononuclear cells. KRG significantly decreased the systolic BP, brachial ankle PWV, and heart femoral PWV in the patients (all p<0.05), but did not significantly alter the serum lipid profiles, including triglycerides and total, high-density lipoprotein, and low-density lipoprotein cholesterol levels. The ROCK activity tended to decrease (p=0.068) following KRG treatment. The placebo did not significantly alter any of the variables. In conclusion, KRG decreased systolic BP and arterial stiffness, probably via the inhibition of ROCK activity, in patients with CAD, but had a neutral effect on serum lipid profiles. Our data suggest that KRG has a therapeutic effect on CAD.

The Study on the Reactivity of Dioxygen Bridged Palladium Complexes Having ${\pi}$-Allyl Ligands (${\pi}$-알릴리간드를 갖는 산소가교 팔라듐착화합물의 반응성에 관한 연구)

  • Pyeong Jin Chung
    • Journal of the Korean Chemical Society
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    • v.30 no.6
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    • pp.516-520
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    • 1986
  • This study is related to reactivity of dioxygen bridged palladium complexes having ${\pi}$-allyl ligands. In this case, new dioxygen bridged palladium complexes were prepared using superoxide ion$(O_2^-)$ as an oxygen source. Reactions of the dioxygen palladium complexes prepared in the study were examined in order to clarify the nature of the coordinated dioxygen. Treatments of a solution of the dioxygen bridged palladium complexes in benzene by water, methanol and acetic acid gave hydrogen peroxide $(H_2O_2)$ as hydroxy-, methoxy-, and acetoxybridged palladium complexes, respectively. The dioxygen bridged palladium complexes reacted also with substitution phenols of salicylaldehyde, 8-hydroxyquinoline and active methylenes of acetylacetone, dimethyl malonate to afford mononuclear complexes of palladium and hydrogen peroxide. The results suggest that dioxygen is coordinated as peroxo $(O_2^{2-})$ in the complexes and behaves as a strong base.

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Epidemiological study on prevalence of fascioliasis in slaughtered cattle (도축 소의 간질 감염실태에 관한 역학적 고찰)

  • Kim, Yong-Hwan;Park, Jong-Tae;Kim, Jae-Ik;Kim, Hyun-Joong;Kim, Tae-Soon;Jang, Mi-Sun;Kim, Jeong-Nam;Park, Duk-Woong;Bae, Seong-Yeol;Park, Ja-Yoon;Seo, Eun-Ju
    • Korean Journal of Veterinary Service
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    • v.31 no.2
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    • pp.207-217
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    • 2008
  • This study was carried out to investigate the prevalence of fascioliasis of slaughtered and farmed cattle in Gwangju area from February to November in 2007. A total of 1,000 cattle fecal samples were collected from slaughter houses (n=805) and farms (n=185). Twelve (1.2%) were found as positive cases with excretion of the egg of Fasciola spp in the fecal specimens, and 128 (12.8%) were positive in intestinal parasitism using the flotation and sedimentation procedures. The infection rate of fluke larvae from the slaughtered cattle at abattoirs in Gwangju was 0.75% (6 out of 805 heads). In histopathology, there were several liver lesions such as inflammation with infiltration of eosinophil, polymorphonuclear cells, mononuclear cells and multinucleated giant cells, proliferation of connective tissue, calcification and abscess formation.