• BMB Reports
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• 제40권4호
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• pp.604-609
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• 2007

An Asp/His catalytic site of 10-formyltetrahydrofolate dehydrogenase (FDH) was suggested to have a similar catalytic topology with the Asp/His catalytic site of serine proteases. Many studies supported the hypothesis that serine protease inhibitors can bind and modulate the activity of serine proteases by binding to the catalytic site of serine proteases. To explore the possibility that soybean trypsin inhibitor (SBTI) can recognize catalytic sites of FDH and can make a stable complex, we carried out an SBTI-affinity column by using rat liver homogenate. Surprisingly, the Rat FDH molecule with two typical liver proteins, carbamoyl-phosphate synthetase 1 (CPS1) and betaine homocysteine S-methyltransferase (BHMT) were co-purified to homogeneity on SBTI-coupled Sepharose and Sephacryl S-200 followed by Superdex 200 FPLC columns. These three liver-specific proteins make a protein complex with 300 kDa molecular mass on the gel-filtration column chromatography in vitro. Immuno-precipitation experiments by using anti-FDH and anti-SBTI antibodies also supported the fact that FDH binds to SBTI in vitro and in vivo. These results demonstrate that the catalytic site of rat FDH has a similar structure with those of serine proteases. Also, the SBTI-affinity column will be useful for the purification of rat liver proteins such as FDH, CPS1 and BHMT.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7335-7338
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• 2013

There is increasing evidence that natural killer (NK) cells play an important role in antitumor immunity following dendritic cell (DC) vaccination. Little is known, however, about the optimal stimulation of DCs by epitopes and NK interactions for cytotoxicity in tumors. In this study, DC cells activated by the HPV16E7.49-57 epitope and LPS were co-cultured with NK cells in vitro, and then used ot immunize mice to study CTL activity of TC-1, which constitutively expresses HPV16E6E7, with an LDH release assay. Cytotoxicity in mice immunized with DC loaded with epitope HPVE7.49-57 vaccine co-cultured with NK was enhanced significantly (p<0.01). In conclusion, talk-across between DC and NK cells enhances their functions, also improving cytotoxicity againsttumor cells, suggesting that activated DC-NK by epitopes has potential application for cancer-specific immuno-cellular therapy.

• 한방안이비인후피부과학회지
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• 제18권1호
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• pp.104-115
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• 2005

Sunjeonhwadok-Tang was a drug that treated carbuncle and cellulitis. So, the purpose of this study was to investigate effects of Sunjeonhwadok-Tang on the proliferation of cancer calls and immunocytes focusing around combined effects of anticarcinogen. We used Sunjeonhwadok-Tang extract(SHT) with freeze-dried, 8wks-old male balb/c mice and cancer cell lines(L1210, Sarcoma-180) for this Study. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results : 1. SHT was significantly showed cytotoxicity on the L1210 cell lines. 2. SHT was significantly increased proliferation of thymocytes and splenocytes in vitro. 3. In combined effects of SHT and vincristine(0.005 mg/kg), SHT was significantly inhibited proliferation of L1210 cell lines, but was not inhibited proliferation of Sarcoma-180 cell lines compared with positive control group. 4. In combined effects of SHT and vincristine(0.005 mg/kg), SHT was significantly increased proliferation of thymocytes and splenocytes compared with positive control group. 5. In combined effects of SHT and vincristine, SHT was significantly increased proliferation of thymocytes and splenocytes in normal mice. 6. In combined effects of SHT and vincristine, SHT was significantly inhibited proliferation of L1210 cells in L1210 cells transplanted mice 7. In combined effects of SHT and vincristine, SHT was significantly increased proliferation of L1210 cell in L1210 cells transplanted mice. The present author thought that SHT had action of anti-cancer and immuno-activity, and in combined effects of vincristine, SHT had recoverable effects on damage by anticarcinogen.

• 대한한의학방제학회지
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• 제17권1호
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• pp.175-185
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• 2009

Petiolus Nelumbinis, branches of lotus leaf or lotus flower is a traditional oriental herbal medicine widely used for treating a superheat or disorder of qi flow. Although there are many clinical results and literature study, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of PNM (Petiolus Nelumbinis MeOH extract) in vitro. To determine cytotoxic concentration of PNM, the cells were treated with PNM for 24 h after LPS addition, and the cell viability was tested by MTT assay. Both of dosages (30 and 100 ${\mu}g/ml$) of PNM had no cytotoxicity. In these concentrations, PNM significantly reduced the elevated levels of NO and $PGE_2$ by LPS. These inhibitory effects of PNM were due to the reduced expressions of iNOS and COX-2 protein. TNF-$\alpha$, IL-1$\beta$ and IL-6 are frequently encountered pro-inflammatory cytokines, and LPS plays a key role in inducing to the massive production of these cytokines. Thus, we next determined the levels of these cytokines. Although PNM had no significant inhibitory effect on the production of TNF-$\alpha$, the elevated levels of IL-1$\beta$ and IL-6 by LPS were dose-dependently reduced in PNM-treated groups. These results demonstrate that PNM has anti-inflammatory effects by inhibiting the production of proinflammatory cytokines, NO and $PGE_2$ in LPS-activated macrophage. Moreover, the reduction of NO and $PGE_2$ levels was due to the inhibition of iNOS and COX-2 protein expression by PNM.

• 대한한방종양학회지
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• 제6권1호
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• pp.125-148
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• 2000

At here, we investigated the historical background, current stage, and concrete therapies of alternatives in cancer therapy. We obtained the conclusion below. 1. Alternative medicine(AM) is a unothodoxial medical physiology, medical practice or intervension that overcome the side effects, faults, limits of coventional medicine. 2. Recentely, as emerging New-age movement, collapse of Decarte's mecanical cosmology, and emphasis on patient rights, the interest of AM is increased, and foreign & domestic use rates of AM are risen. 3. After coventional medicine in cancer therapies that depends on operations, radiations, and chemotherapy is bounded, Alternatives in cancer therapy are rapidly developed, and the majority of users are often considered satisfactory. 4. Alternatives in cancer therapy are classified six boundaries; Diet and Nutrition, Mind-body Technique, Integrated System, Pharmacologic and Biologic treatments, Immuno Enhanced Therapy, Natural and herbal medicine. 5. Looking at contents of Alternatives in cancer therapy, we get to know most of them are similar to the treatment of oriental medicine in holistic method, therapy, philosophy. If we synthesize the alternative medicine's treatment mentioned above and investigate it with oriental medical viewpoint, that may help us approach the conquest of cancer and improve the rate of cure. 6. It is certain that foreign AM will be imported reversely, and we must prepare for rebuilding of Korean traditional AM systemically, investing for activity positively, making national policy for medical system.

• 동의생리병리학회지
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• 제25권1호
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• pp.109-114
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• 2011

Atractylodis Rhizoma Alba is commonly used herbal medicine and it has been known that has immuno-regualtory effects and anti-cancer effects. The inhibition of osteoclastogenesis is essential for the prevention and treatment of osteoporosis. The aim of this study was to evaluate the effects of Atractylodis Rhizoma Alba on osteoclast differentiation in vitro and on resorbing activity of osteoclast. Osteoclast formation was evaluated in bone marrow cells (BMC) in the presence or absence of Atractylodis Rhizoma Alba. The expression of c-fos, tartrate-resistant acid phosphatase (TRAP), OSCAR, DC-STAMP, cathepsin K, MafB and NFATc1 mRNA in osteoclast precursor were assessed by RT-PCR. The levels of TNF receptor-associated factor-6 (TRAF-6), c-fos and NFATc1 protein were assessed by Western blot analysis. Also the correlation with MAPKs and NF-${\kappa}B$ pathways were measured by using Western blot analysis. With bone resorption study, I tried to evaluate the inhibitory effects of Atractylodis Rhizoma Alba on mature osteoclast function. Atractylodis Rhizoma Alba inhibited the RANKL induced osteoclastic differentiation from bone marrow macrophage in a dose dependant manner without cellular toxicity. Gene expression of c-fos and NFATc1 was significantly down regulated with Atractylodis Rhizoma Alba treatment. Atractylodis Rhizoma Alba markedly inhibited the RANKL-induced osteoclastogenesis through suppression of nuclear factor kappa b (NF-${\kappa}B$) pathway, down stream pathway of p38, ERK and JNK pathway. Taken together, I concluded that Atractylodis Rhizoma Alba have beneficial effect on osteoporosis by inhibition of osteoclast differentiation and by inhibition of functioning osteoclast. Thus I expect that Atractylodis Rhizoma Alba could be a treatment option for osteoporosis.

• KSBB Journal
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• 제25권1호
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• pp.47-54
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• 2010

The mycelial dispersed growth of Cordyceps sinensis was optimized in submerged batch culture at initial pH of 5.0, 150 rpm, and $25^{\circ}C$. The morphological data showed much more dispersed growth of C. sinenesis at initial pH of 5.0. Also, projected area, main hyphal length and number of tips for the mycelial growth of initial pH 5.0 were higher than those of other initial pHs. The industrial medium for mycelial production of C. sinensis was determined to be molasses of 100 g and crushed brewery yeast of 10 g per liter as carbon and nitrogen sources, respectively. With these culture conditions, the maximum production of mycelia was approximately 30.0 g per liter by batch culture in 5-liter jar fermenter with no controlled pH. This result suggests that large-scale mycelia production of C. sinensis may be possible in submerged batch culture. The hot water extract of mycelia from C. sinensis was mainly composed of 83.0% carbohydrate, 11.8% protein, 1.9% lipid, and 2.4% ash and there were present glucose, mannose, galactose, and arabinose as molar ratio of 8.79 : 2.59 : 1.34 : 1.0 in the carbohydrate, respectively. In the experiment using spleen cell and macrophage, the extract showed potent mitogenic and immuno-stimulating activities and among various components, an important factor that contribute to the immunological activities was turned out to be carbohydrate moiety.

• 대한한방내과학회지
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• 제24권1호
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• pp.75-83
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• 2003

Objectives : Peroxynitrite($ONOO^-$), formed from the reaction of $O2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been involved in the aging process and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_{2^-}$ and its scavenging mechanism of Zingiberis Rhizoma (ZR). Methods : To investigate scavenging activities of $ONOO^-,\;NO,\;O_{2^-}$ and its scavenging mechanism, we used fluorescent probes like DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on ZR was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. The scavenging efficacy was expressed as IC50, showing the concentration of each sample that is required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of ZR on $ONOO^-$-induced nitration of bovine serum albumin was investigated through immuno-assay with a monoclonal anti-nitryrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results : ZR markedly scavenged authentic $ONOO^-,\;O_{2^-}$ and NO. It also inhibited $ONOO^-$ induced by $O_{2^-}$ and NO which are derived from SIN-1. The data demonstrated that ZR led to decreased $ONOO^-$ mediated nitration of tyrosine through electron donation. It also inhibited the nitration of bovine serum albumin induced by $ONOO^-$ in a dose-dependent manner. Furtheremore, it blocked LPS-induced ROS and RNS generation. Conclusions : These results suggest that ZR can be developed as an effective $ONOO^-$ scavenger for the prevention of aging process and age-related diseases.

• 대한임상검사과학회지
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• 제40권1호
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• pp.31-40
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• 2008

Despite of the importance of the primordial follicle (PMF) recruitment, factors and mechanisms for process are poorly understood. To evaluate expression and role of the follicular transition from PMF to PMF/primary follicles (PMIF) in the present study, we evaluated expression of lats1, lats2, cyclin A1, and cyclin A2 mRNA and protein, and elucidated and role of lats1-cyclin A in the follicular transition from PMF to PRIF. To analysis of differential expression in PMF and PMIF, each stage follicles were collected by day1 and day5 of immuno-compromised rats (ICR) and analyzed by real-time PCR for the genes. For localization of mRNAs and proteins of the genes, in situ hybridization and immunohistochemistry were performed. We confirmed that the lats1, lats2, cyclin A1, and cyclin A2 mRNA were more expressed in PMF than PMIF. Localization of the four genes expression were observed in nuclei of oocytes from the arrested primordial, and in the surrounding granulosa cells of the growing follicles. The mRNA expressions were gradually decreased with follicular development. From immunohistochemistry studies, Cyclin A1 protein expression were observed in oocyte cytoplasmas of early stage follicles, while observed in granulose cells and oocyte nucleoli during growing follicles. This study suggested that the presence of lats gene family might perform negatively regulation of cell proliferation by modulation of the CDC2/Cyclin A complex activity. lats-cyclin A genes in oocytes of the early stage follicles might play a role in the meiotic cell cycle arrest of the primary oocytes at the primordial follicle stage as well as the follicular growth.

• 동의생리병리학회지
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• 제24권6호
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• pp.956-961
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• 2010

Tumor necrosis factor-alpha (TNF-${\alpha}$) is a major mediator of immuno-inflammatory activity. The purpose of this study is to investigate whether TNF-${\alpha}$ productions of mouse macrophage RAW 264.7 and human hepatocyte HepG2 are modulated by Artemisiae argi Folium water extract (AW), Lactobacillus pentosus-fermented Artemisiae argi Folium water extract (AFL), and Saccharomyces cerevisiae-fermented Artemisiae argi Folium water extract (AFS) for 3 h of incubation. Effect of AW on cell viability of HepG2 was also investigated. TNF-${\alpha}$ productions were measured by Enzyme-Linked Immnunosorbent Assay method and cell viability was measured by MTT assay. Both AFL and AFS significantly increased TNF-${\alpha}$ productions of RAW 264.7 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). Also, AFL and AFS significantly increased TNF-${\alpha}$ productions of HepG2 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). AW significantly increased TNF-${\alpha}$ production of HepG2 at the concentration of 100 and 200 ${\mu}g$/mL (p<0.05). AW did not show any cytotoxicity on HepG2 cells for 3 h. These results suggest that AFL, AFS, and AW have the immune-enhancing property related with its increasing effect on TNF-${\alpha}$ production of macrophage and hepatocyte.