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Vasorelaxing Mechanism of Crude Saponin of Korea Red Ginseng in the Resistance-sized Mesenteric Artery of Rat

  • Kim, Shin-Hye;Park, Hyung-Seo;Lee, Mee-Young;Oh, Young-Sun;Kim, Se-Hoon
    • Journal of Ginseng Research
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    • v.26 no.1
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    • pp.1-5
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    • 2002
  • It has been well known that Korea red ginseng has an antihypertensive effect. The antihypertensive effect may be due to its ability to change the peripheral resistance. Change of vascular tone in the resistance-sized artery contribute to the peripheral resistance, thereby regulate the blood pressure. Therefore, we investigated to clarify the vasorelaxing mechanism induced by crude saponin of Korea red ginseng in the resistance-sized mesenteric artery of rats. The resistance-sized mesenteric artery was isolated and cut into a ring. The ring segment was immersed in HEPES-buffered solution and its isometric tension was measured using myograph force-displacement transducer. Crude saponin of ginseng relaxed the mesenmetric arterial rings precontracted with norepinephrine (3$\mu$M) in dose-dependent manner (0.01 mg/㎖ -1 mg/㎖. The relaxation by crude saponin was smaller in endothelium-intact preparation than that in endothelium-denuded preparation. The contraction induced by A23187 or phorbol 12,13-dibutyrate was not affected by crude saponin of ginseng. The vasorelaxing effect of crude saponin of ginseng was significantly attenuated by the increase of the extracellular K$\^$+/ concentration. Crude saponin-induced vasorelaxation was not affected by tetraethylammonium (1 mM), glybenclamide (10$\mu$M), and 4-aminopyridine (0.1 mM) in these preparations. Ba$\^$2+/(10$\mu$M ∼100$\mu$M) markedly reduced the crude saponin-induced vasorelakation dose-dependently. From the above results, we suggest that crude saponin of ginseng may stimulate K$\^$+/ efflux and hyperpolarize the membrane, thereby cause the vasorelaxation in the resistance-sized mesenteric artery of rats.

Inhibitory Effects of Panaxatriol from Panax ginseng C. A. Meyer on Phosphoinositide Breakdown Induced by Thrombin in Platelets

  • Park, Kyeong-Mee;Rhee, Man-Hee;Shin, Han-Jae;Song, Yong-Bum;Hyun, Hak-Chul;Park, Ki-Hyun;Cho, Hyun-Jeong;Choi, Sun-A;Kang, Hyo-Chan;Kim, Kyoung-Jin;Kim, Hyeong-Soo;Kang, Hee-Jin;Ok, Woo-Jeong;Lee, Dong-Ha;Park, Hwa-Jin
    • Journal of Ginseng Research
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    • v.32 no.2
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    • pp.107-113
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    • 2008
  • In this study, we have investigated the effect of panaxatriol (PT) on phosphoinositides (PIS) breakdown and $Ca^{2+}$-elevation in thrombin-induced platelet aggregation. Thrombin (5U/ml), a potent platelet agonist which activates phospholipase $C_{\beta}$ via protease activated receptor (PAR), hydrolyzed PIS in platelet membrane. The phosphatidylinositol 4, 5-bisphosphate $(PIP_2)$ was hydrolyzed after 10 sec of the thrombin-stimulation, and both the phosphatidylinositol 4-monophosphate (PIP) and phosphatidylinositol (PI) were brokendown after 30 sec of the thrombin-stimulation. However, PT inhibited the thrombin-stimulated hydrolysis of $PIP_2$, PIP, and PI. On the other hand, thrombin increased the level of phosphatidic acid (PA) which is phosphorylated from diacylglycerol (DG) generated by PIS-hydrolysis. However, Pr inhibited the thrombin-increased PA level non-significantly. Thrombin increased cytosolic free $Ca^{2+}([Ca^{2+}])_i$) up to 72% as compared with control $(30.8{\pm}0.9 nM)$ in intact platelet. However, PT (100 ${\mu}g/ml$) inhibited the thrombin-elevated $[Ca^{2+}]_i$ to 100%. These results suggest that PT may have a beneficial effect on platelet aggregation-mediated thrombotic disease by inhibiting thrombin-induced platelet aggregation via suppression of the $[Ca^{2+}]_i$ level and PIS breakdown.

Combined Treatment of Sodium Salicylate and Genistein Induces Incomplete Apoptosis and Necrosis in MCF-7 Multicellular Tumor Spheroids (MCF-7 MTS에서 sodium salicylate과 genistein 복합처리는 불완전한 세포사멸과 세포괴사를 유도한다)

  • Lee, Su-Yeon;Kim, Cho-Hee;Jeon, Hyun-Min;Ju, Min-Kyung;Kim, Min-Young;Jeong, Eui-Kyong;Park, Hye-Gyeong;Kang, Ho-Sung
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1145-1151
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    • 2012
  • Aspirin and its deacetylated form, sodium salicylate (NaSal), have been shown to exert chemopreventive activities against many human cancers including those of the colon, lung, and breast. Previously, we showed that combined treatment of NaSal and genistein synergistically induced apoptosis in A549 lung cancer cells, indicating that these two natural chemicals could be used in combination for cancer therapy. In this study, we examined effects of NaSal/genistein combined treatment on other cancer cells and in three-dimensional multicellular tumor spheroid (MTS) and in an in vitro solid tumor model. We found that the combined treatment induces apoptosis in the HCT116 cells and the A549 cells, but not in the MCF-7 cells. Interestingly, the MCF-7 cells responded to the NaSal/genistein combined treatment by undergoing cell death when they were cultivated as MTS. The combined treatment induced apoptosis at an earlier stage in the MCF-7 MTS culture. However, when the MCF-7 MTS was cultivated for a longer period, it induced necrosis rather than apoptosis. We further found that the apoptotic pattern observed in MCF-7 MTS was incomplete: the chromatins were condensed and fragmented, but the nuclear membrane was still intact. Taken together, these results demonstrate that the NaSal/genistein combined treatment induces incomplete apoptosis and necrosis in the MCF-7 MTS culture system.

Induction of Apoptosis in HT-29 Human Colon Cancer Cells by the Pepper Component Piperine (후추의 주요 성분인 Piperine의 대장암세포 세포사멸 유도 효과)

  • Kim, Eun-Ji;Park, Hee-Sook;Shin, Min-Jeong;Shin, Hyun-Kyung;YoonPark, Jung-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.442-450
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    • 2009
  • Piperine is an alkaloid-amine found in pepper and has been reported to have anticarcinogenic properties. To explore the possibility that piperine has cancer chemopreventive and chemotherapeutic effects in colon cancer, we examined whether piperine inhibits the growth of HT-29 human colon cancer cells and investigated the mechanisms for this effect. Cells were cultured with various concentrations ($0{\sim}40{\mu}M$) of piperine. Piperine decreased the cell viability and induced apoptosis of HT-29 cells. Western blot analysis of total cell lysates revealed that piperine decreases the protein levels of Bcl-2, Mcl-1, and intact Bid but increases Bik levels. Piperine increased the percentage of cells with depolarized mitochondrial membrane, and the release of cytochrome c into cytoplasm. Piperine induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3 and increased the Fas levels. In addition, piperine significantly decreased the protein levels of survivin. The present results indicate that piperine inhibits the growth of HT-29 colon cancer cells by the induction of apoptosis, which may be mediated by its ability to change the Bcl-2 family proteins, increase the activation of caspases, and decrease survivin levels. Overall, our findings suggest that piperine has cancer chemotherapeutic effects in colon cancer.

Treatment of Aplasia Cutis Congenita on Scalp using Hyalomatix$^{(R)}$: A Case Report (하이알로매트릭스를 이용한, 두개골결손을 동반한 선천성피부무형성증의 치료: 1례 보고)

  • Rhee, Suk-Hyun;Hong, Jong-Won;Roh, Tai-Suk;Kim, Young-Seok;Rah, Dong-Kyun
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.469-472
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    • 2010
  • Purpose: Aplasia Cutis Congenita (ACC) is a rare disease characterized by the focal defect of the skin at birth, frequently involving scalp, but it may affect any region of the body. There are no etiology known but some conditions such as intrauterine vascular ischemia, amniotic adherences and viral infections are associated. The ideal treatment for the ACC is not known. Superficial and relatively small sized defects (< $3{\times}5\;cm$) may heal spontaneously and large defects related with risks of infection and bleeding may require aggressive surgical treatment. Hyalomatrix$^{(R)}$ is a bilayer of an esterified hyaluronan scaffold beneath a silicone membrane. It has been used as a temporary dermal substitute to cover deep thickness skin defect and has physiological functions derive from the structural role in extracellular matrix and interaction with cell surface receptor. This material has been used for the wound bed pre-treatment for skin graft to follow and especially in uncooperative patient, like a newborn, this could be a efficient and aseptic way of promoting granulation without daily irritative wound care. For this reason, using Hyalomatrix$^{(R)}$ for the treatment of ACC was preferred in this paper. Methods: We report a case of a newborn with ACC of the vertex scalp and non-ossified partial skull defect. The large sized skin and skull defect ($6{\times}6\;cm$) was found with intact dura mater. No other complications such as bleeding or abnormal neurologic sign were accompanied. Escharectomy was performed and Hyalomatrix$^{(R)}$ was applied for the protection and the induction of acute wound healing for 3 months before the split-thickness skin graft. During the 3 months period, the dressing was renewed in aseptic technique for every 3 weeks. The skin graft was achieved on the healthy granulation bed. Results: The operative procedure was uneventful without necessity of blood transfusion. Postoperative physical examination revealed no additional abnormalities. Regular wound management was performed in out-patient clinic and the grafted skin was taken completely. No other problems developed during follow-up. Conclusion: Hyalomatrix$^{(R)}$ provides protective and favorable environment for wound healing. The combination of the use of Hyalomatrix$^{(R)}$ and the skin graft will be a good alternative for the ACC patients with relatively large defect on vertex.

Inactivation of Pathogenic Escherichia coli Using Crude Extract of Immunized Silkworm (면역유도누에 추출물을 이용한 병원성 대장균의 불활성화)

  • Park, Jong Woo;Jeong, Chan Young;Lee, Chang Hoon;Kang, Sang Kuk;Ju, Wan-Taek;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young
    • Journal of Life Science
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    • v.31 no.8
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    • pp.755-760
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    • 2021
  • Swine diarrhea is a livestock disease that causes huge economic losses to pig farms. In general, diarrhea occurs because of the proliferation of pathogenic Escherichia coli (E. coli). The toxins produced by the proliferated E. coli cause edema in pigs. Although the proliferation of these coliforms can be prevented by using a vaccine, the vaccines containing chemically produced dead bacteria are not very effective, making it difficult to control the proliferation of E. coli. Therefore, there is a need to develop new, more effective vaccines. In this study, we prepared killed F4+ and F18ab+ E. coli, which induce diarrhea and edema in pigs, using the extracts of immune-induced silkworms containing antimicrobial peptides and examined their availability as a killed-bacteria vaccine. First, the antimicrobial activity analysis of the prepared immune-induced silkworm extract was conducted using the radial diffusion assay. The results showed high activity against both F4+ and F18ab+ E. coli. The production efficiency of E. coli dead cells was determined using the colony-counting method. The concentration of the E. coli dead cells was the highest (50 mg/ml) when treated at 4℃. In addition, the analysis of the prepared dead cells using a transmission electron microscope confirmed that E. coli leaked out of the cytoplasm and the cell membrane remained intact. Therefore, F4+ and F18ab+ E. coli produced using immune-induced silkworms extract are considered to be highly available as bacterial ghost vaccines that can help prevent swine diarrhea and the resulting edema.

Monitoring trafficking and expression of hemagglutinin-tagged transient receptor potential melastatin 4 channel in mammalian cells

  • Eun Mi Hwang;Bo Hyun Lee;Eun Hye Byun;Soomin Lee;Dawon Kang;Dong Kun Lee;Min Seok Song;Seong-Geun Hong
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.4
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    • pp.417-426
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    • 2023
  • The TRPM4 gene encodes a Ca2+-activated monovalent cation channel called transient receptor potential melastatin 4 (TRPM4) that is expressed in various tissues. Dysregulation or abnormal expression of TRPM4 has been linked to a range of diseases. We introduced the hemagglutinin (HA) tag into the extracellular S6 loop of TRPM4, resulting in an HA-tagged version called TRPM4-HA. This TRPM4-HA was developed to investigate the purification, localization, and function of TRPM4 in different physiological and pathological conditions. TRPM4-HA was successfully expressed in the intact cell membrane and exhibited similar electrophysiological properties, such as the current-voltage relationship, rapid desensitization, and current size, compared to the wild-type TRPM4. The presence of the TRPM4 inhibitor 9-phenanthrol did not affect these properties. Furthermore, a wound-healing assay showed that TRPM4-HA induced cell proliferation and migration, similar to the native TRPM4. Co-expression of protein tyrosine phosphatase, non-receptor type 6 (PTPN6 or SHP1) with TRPM4-HA led to the translocation of TRPM4-HA to the cytosol. To investigate the interaction between PTPN6 and tyrosine residues of TRPM4 in enhancing channel activity, we generated four mutants in which tyrosine (Y) residues were substituted with phenylalanine (F) at the N-terminus of TRPM4. The YF mutants displayed properties and functions similar to TRPM4-HA, except for the Y256F mutant, which showed resistance to 9-phenanthrol, suggesting that Y256 may be involved in the binding site for 9-phenanthrol. Overall, the creation of HA-tagged TRPM4 provides researchers with a valuable tool to study the role of TRPM4 in different conditions and its potential interactions with other proteins, such as PTPN6.

Selectivity of Bleaching Herbicides Caused by Physiological Differences between Rice and Barnyardgrass (벼와 피의 생리적(生理的) 차이(差異)에 의한 백화형(白化型) 제초제(除草劑)의 선택성(選擇性))

  • Na, J.Y.;Kim, J.S.;Kim, T.J.;Cho, K.Y.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.89-101
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    • 1992
  • In this study, various physiological and biochemical experiments were conducted to know whether the selectivity between rice and barnyardgrass treated with bleaching herbicides containing diphenyl ether compounds was also partly based on their basic physiological proterties such as peroxidation ability, membrane stability or antioxidant system. It seemed to be partly based on the differences of their physiological characteristics that barnyardgrass was commonly more susceptible to most of the bleaching herbicides than rice. The scenescence of intact leaf segment was more rapid in barnyardgrass than in rice, indicating that barnyardgrass is weak at early stage. Also pigment metabolic ability, antioxidant enzyme activities(peroxidase, catalase, superoxide dismutase, glutathione reductase) and antioxidant content (tocopherol, ascorbic acid, glutathione, carotenoids) were lower in barnyardgrass on the basic of fresh weight. However, lipoxygenase activity and the content of unsaturated fatty acid which is vulnerable to oxygen radicals were higher in barnyardgrass, suggesting that barnyardgrass seedling bave a properties easy to be peroxidized. The differences of PPIX (protoporphyrin IX) or carotenoid content, which are the primary substances inducing herbicide activity, were not related to the selectivity between rice and barnyardgrass.

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Effects of Green Tea on Weight Gain, Plasma and Liver Lipids and Lipid Peroxidation in Pair Fed Rats (Pair Fed 흰쥐에 있어서 녹차의 항증체, 지질개선 및 항산화 효과)

  • Kang, Jung-Ae;Chae, In-Sook;Song, Yong-Bo;Kang, Jung-Sook
    • Journal of Nutrition and Health
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    • v.41 no.7
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    • pp.602-611
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    • 2008
  • We compared antiobese, hypocholesterolemic, antiplatelet and antioxidant effect of 10% green tea powder and 3% green tea extract in rats pair fed 5% cholesterol diets. The final body weight was decreased significantly compared with the control (p < 0.05). Plasma and liver total cholesterol were lower in group of green tea powder or extract, but not statistically different. HDL cholesterol was increased significantly in group of green tea powder compared with the control or green tea extract (p < 0.05). Plasma triglyceride was significantly decreased in group of green tea extract compared with green tea powder, and green tea powder compared with the control respectively (p < 0.05). Liver triglyceride was significantly decreased in group of green tea powder or green tea extract compared with the control (p < 0.01). Platelet aggregations in the maximum and initial slope were not different among groups. Hemolysis was significantly lower in group of green tea powder compared with the control (p < 0.05). Plasma TBARS production was decreased in group of green tea extract compared with the control (p < 0.05). Na passive leak in intact cells was not different, but Na leak in AAPH treated cell was significantly decreased in group of green tea powder than the control (p < 0.05). The leak increase (${\Delta}Na$ Leak) after AAPH treatment was significantly decreased in groups of green tea powder and extract compared with the control (p < 0.05). Isotope excretion after $^{14}C$-cholesterol ingestion was significantly increased in group of green tea extract compared with the control or the green tea powder (p < 0.05). Consumption of green tea in powder or extract may give beneficial effects in weight control and plasma lipid profiles, impeding metabolic syndrome. More studies are needed to clarify what component of green tea and what mechanism are involved in antiobese and hypolipedemic actions of green tea.

The Study of CYFRA 21-1 and Epidermal Growth Factor Receptor Levels in Cancer Tissue of Bronchogenic Carcinoma Patients (폐암환자의 암조직내 CYFRA 21-1과 Epidermal Growth Factor Receptor의 측정치에 대한 연구)

  • 김대연;김송명
    • Journal of Chest Surgery
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    • v.30 no.9
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    • pp.854-861
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    • 1997
  • CYPRA 21-1 is known to be a cytokeratin 19 fragment, and it can be detected by using two specific monoclonal antibodies (KS 19-1 and BM 19-21) and can be clinically applied as a useful circulating tumor marker The epidermal growth factor receptor (EGF-R) expression was evaluated and characterized by its tyrosine protein kinase activity and by its ligand-stimulated autophosphorylation, a property shared with other peptide growth factor receptors. Autocrine or para'urine action was initiated by a growth factor, or by a transforming growth factor o, which had an extensive homology with EGP and which also stimulated tyrosine kinase activity on the EGF-R. The CYFRA 21-1 and the EGF-R levels in 30 patients with primary lung tumors were investigated. There were 24 patients with squamous cell carcinomas and 6 patients with adenocarcinomas. Specimen 5 mm3 in size were sampled at three different locations ; the main lesion, the boundary between the lesion and the unaffected tissue, and the unaffected tissue of the patients. The results were as follows 1. The CYPRA 21-1 concentration in the cancer boundary, the most malignant region,(348.6 : 89.9 ng/ml) was the lowest value. The CYFRA 21-1 concentration in unaffected tissue,(718.4$\pm$77.8 ng/ml) was higher than that in the main lesion. which had intact cellularity. 2. The EGF-R concentration in the main lesion was higher than that in the unaffected tissue, and the EGF-R concentration in a squamous cell cacinoma was higher than that in an adenocarcinoma. also, the EGF-R concentration in the cancer b undary was highest at stage 1, ll. The EGF-R concentration was higher in the main cancer lesion that in the unaffected tissue at stage 111, IV. 3. The CYFRA 21-1 was a cytoplasmic skeleton and the EGF-R was a cell-wall component; there was no correlation. In conclusion, CYFRA 21-1 was abundant in the cytoplasm but had a higher concentration in the unaffected tissue than in the main cancer lesion. The CYFRA 21-1 concentration of the tissue did not reflect the amount of cancer activity, the EGP-R was located in the cell membrane, the level of tissue that reflects cancer activity, so the main cancer lesion had a higher concentration than the unaffected tissue. CYFRA 21-1 is not a useful tumor maker at the tissue level. Because the EGF-R concentration re(looted the cancer activity, its a useful tumor marker for lung cancer.

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