• The Journal of Korean Medicine
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• v.34 no.3
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• pp.69-85
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• 2013

Objectives: This study investigated the anti-osteoarthritic effects of Kyejiinsam-tang (hereinafter referred to KIT) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods: Anti-oxidative effects of KIT were measured by scavenging activities of DPPH, reactive oxygen species (ROS) and nitric oxide (NO). Scavenging activities of anti-oxidation in lipopolysaccharide (LPS)-treated RAW 264.7 cells were also measured for inhibitory effects against the production of inflammatory mediators (tumor necrosis factor-${\alpha}$, interleukin-$1{\beta}$, interleukin-6). Osteoarthritis was induced in rats by injecting MIA in the knee joint. Rats were divided into a total of 4 groups (n=6). The normal group were not treated at all without inducing osteoarthritis whereas the control group were induced for osteoarthritis by MIA and oral medicated physiological saline per day. The positive comparison group was injected with MIA and after 7 days, 2 mg/kg of Indomethacin. The experimental group was injected with MIA and after 7 days was medicated with 34 mg/kg of KIT. Indomethacin and KIT were orally-medicated for each substance a total of 4 weeks, once per day. Weight-bearing on hind legs was measured every week after MIA injection. At the end of the experiment (5 weeks after MIA injection), micro CT (computed tomography)-arthrography and histopathological examinations on the articular structures of knee joint were performed. The effect on inflammatory cytokines and immunological cells in synovial fluid was measured. Volume of cartilage was measured by micro CT-arthrography. Injury to synovial tissue was measured by H & E (hematoxylin and eosin), Safranin-O immunofluorescence. Results: 1. Cytotoxicity against hFCs was insignificant. 2. KIT showed the potent full term for DPPH. 1. NO was significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and ROS was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 2. IL-6 and IL-$1{\beta}$ were significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and TNF-${\alpha}$ was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 1. In hind legs weight-bearing measurement, level of weight increased. 2. Functions of liver and kidney were not affected. 3. IL-$1{\beta}$ was significantly reduced and TNF-${\alpha}$, IL-6 were also reduced but not significantly. 4. PGE2 (prostaglandin E2), LTB4 (leukotriene B4) were significantly reduced in the KIT group. 5. MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinases-1) and Osteocalcin were significantly reduced in the KIT group. 6. Destruction of cartilage on micro CT arthrography was reduced but had no significant differences. 7. Histopathologically, injury to synovial membrane of the KIT group was decreased and proteoglycan content of KIT group was increased. Conclusions: According to this study, Kyejiinsam-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Kyejiinsam-tang has anti-oxidants and anti-inflammation effects, and is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.9
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• pp.4669-4675
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• 2012

Objective: Nude mice with orthotopic transplantation of human ovarian epithelial cancer were used to investigate screening criteria for paraneoplastic normal ovarian tissue and the security of the freezing and thawing for ovarian tissue transplantation. Methods: Expression of CK-7, CA125, P53, survivin, MMP-2/TIMP-2 in paraneoplastic normal ovarian tissues were detected by RT-PCR as well as immunohistochemistry. The tissues of the groups with all negative indicators of RT-PCR, all negative indicators of immunohistochemistry, negative expression of CK-7, CA125 and survivin, positive expression of CK-7, CA125 and survivin, cancer tissues and normal ovarian tissues of nude mice were used for freezing and thawing transplantation, to analyze overt and occult carcinogenesis rates after transplantation. Results: When all indicators or the main indicators, CK-7, CA125 and survivin, were negative, tumorigenesis did not occur after transplantation. In addition the occult carcinogenesis rate was lower than in the group with positive expression of CK-7, CA125 and survivin (P<0.01). After subcutaneous and orthotopic transplantation of ovarian tissues, rates did not change (P>0.05). There was no statistical significance among rates after transplantation of ovarian tissues which were obtained under different severity conditions (P>0.05). Conclusion: Negative expression of CK-7, CA125 and survivin can be treated as screening criteria for security of ovarian tissues for transplantation. Immunohistochemical methods can be used as the primary detection approach. Both subcutaneous and orthotopic transplantation are safe. The initial severity does not affect the carcinogenesis rate after tissue transplantation. Freezing and thawing ovarian tissue transplantation in nude mice with human epithelial ovarian carcinoma is feasible and safe.

• Molecules and Cells
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• v.38 no.6
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• pp.528-534
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• 2015

Hypoxia-inducible factor (HIF) is a key regulator of tumor growth and angiogenesis. Recent studies have shown that, BIX01294, a G9a histone methyltransferase (HMT)-specific inhibitor, induces apoptosis and inhibits the proliferation, migration, and invasion of cancer cells. However, not many studies have investigated whether inhibition of G9a HMT can modulate HIF-$1{\alpha}$ stability and angiogenesis. Here, we show that BIX01294 dose-dependently decreases levels of HIF-$1{\alpha}$ in HepG2 human hepatocellular carcinoma cells. The half-life of HIF-$1{\alpha}$, expression of proline hydroxylase 2 (PHD2), hydroxylated HIF-$1{\alpha}$ and von Hippel-Lindau protein (pVHL) under hypoxic conditions were decreased by BIX01294. The mRNA expression and secretion of vascular endothelial growth factor (VEGF) were also significantly reduced by BIX01294 under hypoxic conditions in HepG2 cells. BIX01294 remarkably decreased angiogenic activity induced by VEGF in vitro, ex vivo, and in vivo, as demonstrated by assays using human umbilical vein endothelial cells (HUVECs), mouse aortic rings, and chick chorioallantoic membranes (CAMs), respectively. Furthermore, BIX01294 suppressed VEGF-induced matrix metalloproteinase 2 (MMP2) activity and inhibited VEGF-induced phosphorylation of VEGF receptor 2 (VEGFR-2), focal adhesion kinase (FAK), and paxillin in HUVECs. In addition, BIX01294 inhibited VEGF-induced formation of actin cytoskeletal stress fibers. In conclusion, we demonstrated that BIX01294 inhibits HIF-$1{\alpha}$ stability and VEGF-induced angiogenesis through the VEGFR-2 signaling pathway and actin cytoskeletal remodeling, indicating a promising approach for developing novel therapeutics to stop tumor progression.

• IMMUNE NETWORK
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• v.14 no.1
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• pp.45-53
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• 2014

Osteoarthritis (OA) is a degenerative joint disease characterized by a progressive loss of cartilage. And, increased oxidative stress plays a relevant role in the pathogenesis of OA. Ursodeoxycholic acid (UDCA) is a used drug for liver diseases known for its free radical-scavenging property. The objectives of this study were to investigate the in vivo effects of UDCA on pain severity and cartilage degeneration using an experimental OA model and to explore its mode of actions. OA was induced in rats by intra-articular injection of monosodium iodoacetate (MIA) to the knee. Oral administration UDCA was initiated on the day of MIA injection. Limb nociception was assessed by measuring the paw withdrawal latency and threshold. Samples were analyzed macroscopically and histologically. Immunohistochemistry was used to investigate the expression of interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, nitrotyrosine and inducible nitric oxide synthase (iNOS) in knee joints. UDCA showed an antinociceptive property and attenuated cartilage degeneration. OA rats given oral UDCA significantly exhibited a decreased number of osteoclasts in subchondral bone legion compared with the vehicle-treated OA group. UDCA reduced the expression of IL-$1{\beta}$, IL-6, nitrotyrosine and iNOS in articular cartilage. UDCA treatment significantly attenuated the mRNA expression of matrix metalloproteinase-3 (MMP-3), -13, and ADAMTS5 in IL-$1{\beta}$-stimulated human OA chondrocytes. These results show the inhibitory effects of UDCA on pain production and cartilage degeneration in experimentally induced OA. The chondroprotective properties of UDCA were achieved by suppressing oxidative damage and inhibiting catabolic factors that are implicated in the pathogenesis of cartilage damage in OA.

• Journal of Embryo Transfer
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• v.33 no.3
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• pp.119-127
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• 2018

The purpose of this study is to examined the electrofusion and activation conditions for the production of porcine somatic cell nuclear transfer (SCNT) embryos. In this study, immature oocytes were cultured in TCM-199 with and without hormones for 22 hours. Skin fibroblasts cells of porcine were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion was performed with two different pulses that each one pulse (DC) of 1.1 kV/cm or 1.5 kV/cm for $30{\mu}sec$. After fusion subsequent activation were divided into three groups; non-treatment (control) and treatment with 2 mM 6-DMAP or $7.5{\mu}g/ml$ cytochalasin B for 4 hours. Transferred embryos were cultured in PZM-3 (Porcine Zygote Medium-3) in $5%\;CO_2$ and 95% air at $39^{\circ}C$ for 7 day. Apoptosis-related genes (Caspase-3, BCL-2, mTOR, and MMP-2) were analyzed by immunofluorescence staining. There was no significant difference between two different electrofusion stimuli in the cleavage rate; $64.9{\pm}4.8%$ in 1.1 kV/cm and $62.7{\pm}4.0%$ in 1.5 kV/cm. However, blastocyst formation rate (%) was significantly different among three different activation groups (no treatment, 2 mM 6-DMAP or $7.5{\mu}g/ml$ cytochalasin B) combined with electrofusion of 1.1 kV/cm. The blastocyst formation rate was $12.6{\pm}2.5$, $20.0{\pm}5.0$, and $34.9{\pm}4.3%$ in control, 2 mM 6-DMAP, and $7.5{\mu}g/ml$ cytochalasin B, respectively. Immunofluorescence data showed that expression levels of caspase-3 in SCNT embryos undeveloped to blastocyst stage were higher than those in the blastocyst stage embryos. Expression levels of Bcl-2 in blastocyst stage embryos were higher than those in the arrested SCNT embryos. These results showed that the combination of an electric pulse (1.1 kV/cm for $30{\mu}sec$) and $7.5{\mu}g/ml$ cytochalasin B treatment was effective for production of the porcine SCNT embryos.

• The Journal of Internal Korean Medicine
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• v.23 no.2
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• pp.165-173
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• 2002

Purpose : Among numerous biological symptoms of cancer, matrix metalloproteinases (MMPs) are essential for tumor invasion and metastasis. HAD is used as an inhibitor of MMP gene. This study was designed to evaluate the effects of HAD on anti metastasis and preventing recurrence in cancer patients. Materials and Methods : We retrospectively analyzed the medical records of 69 cancer patients who had been administered with HAD for over 12 months continuously in East-West Cancer Center of Oriental Hospital of Daejeon University, from January 1993 to May 2002. Results : We analyzed gender, portion, stage and anti-metastasis & recurrence rates of cancer patients. Analysis of sex cases showed that the percentage of male is 62.3%, female is 37.7%. Analysis of cancer portion showed that the percentage of stomach is 31.9%, colorectum is 26.1%, lung is 21.7%, liver is 8.7%, breast is 8.7% Analysis of stage showed that the rate of III is 78.3%, IV is 13.0% and II is 8.7%. Analysis of anti-metastasis and recurrence rates showed that colorectal cancer is 77.8%, stomach cancer is 63.6%, lung cancer is 33.4% and breast cancer is 33.3% (mean : 53.6%). Conclusions : HAD has significant effects on anti-metastasis and preventing recurrence of tumor on cancer patients. So it helps to prolong the survival rates of cancer patients.

• Toxicological Research
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• v.34 no.2
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• pp.103-110
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• 2018

Environmental stimuli can lead to the excessive accumulation of reactive oxygen species (ROS), which is one of the risk factors for premature skin aging. Here, we investigated the protective effects of $7-MEGA^{TM}$ 500 (50% palmitoleic acid, 7-MEGA) against oxidative stress-induced cellular damage and its underlying therapeutic mechanisms in the HaCaT human skin keratinocyte cell line (HaCaT cells). Our results showed that treatment with 7-MEGA prior to hydrogen peroxide ($H_2O_2$)-induced damage significantly increased the viability of HaCaT cells. 7-MEGA effectively attenuated generation of $H_2O_2$-induced reactive oxygen species (ROS), and inhibited $H_2O_2$-induced inflammatory factors, such as prostaglandin $E_2$ ($PGE_2$), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), and $interleukin-1{\beta}$ ($IL-1{\beta}$). In addition, cells treated with 7-MEGA exhibited significantly decreased expression of matrix metalloproteinase-1 (MMP-1) and increased expression of procollagen type 1 (PCOL1) and Elastin against oxidative stress by $H_2O_2$. Interestingly, these protective activities of 7-MEGA were similar in scope and of a higher magnitude than those seen with 98.5% palmitoleic acid (PA) obtained from Sigma when given at the same concentration (100 nL/mL). According to our data, 7-MEGA is able to protect HaCaT cells from $H_2O_2$-induced damage through inhibiting cellular oxidative stress and inflammation. Moreover, 7-MEGA may affect skin elasticity maintenance and improve skin wrinkles. These findings indicate that 7-MEGA may be useful as a food supplement for skin health.

• Journal of Korean Traditional Oncology
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• v.11 no.1
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• pp.41-54
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• 2006

Ekongsan (EKS) was expected to have inhibitory effects on angiogenesis, considering the fact that its constituents such as Ginseng Radix, Glycyrrhizae Radix and Citri Pericarpium were reported to inhibit angiogenesis. Moreover, recently several metabolites transformed by the human intestinal microflora were reported to enhance effectiveness compared to their crude drugs. Based on these data, this study was designed to confirm whether the EKS metabolites (EKS-M) can significantly exert the anti-angiogenic and anti-metastatic activites. Hence, with EKS and EKS-M, viability assay, proliferation assay, in vitro tube formation assay, gelatin zymogram assay, in vitro invasion assay were carried out. EKS showed less toxicity in ECV304 and HT1080 cells than EKS-M. EKS-M inhibited the proliferation of HT1080 cells by 30% at 200 ${\mu}g/m{\ell}$ and 42% at 400 ${\mu}g/m{\ell}$ respectively. Also, EKS-M degraded the tube network at 200 ${\mu}g/m{\ell}$. EKS and EKS-M inhibited the expression of MMP-9 at 200 and 400 ${\mu}g/m{\ell}$in HT1080 cells. EKS reduced the invasive activity of HT1080 cells through matrigel coated transfilter at the concentration of 200 ${\mu}g/m{\ell}$ more effectively than EKS-M. These data suggest that EKS and EKS-M has anti-angiogenic and anti-metastatic activities.

• Journal of Korean Traditional Oncology
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• v.11 no.1
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• pp.75-93
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• 2006

Jiaweicitaowan (JWCTW) has been used to inhibit recurrence and metastasis of cancer in clinical practice. Further study has shown its anti-metastatic and anti-angiogenic effects. By applying in vitro and in vivo anti-angiogenic evaluation model, the author assayed the each ingredients of JWCTW. The author performed the following experiments and the results are listed as below: Cell viability assay showed that the viability was almost identical between that of the control and that of the ingredients extract 40 ${\mu}g/m{\ell}$ treated, except of hexane fraction of Curcumae Radix (40 ${\mu}g/m{\ell}$, 2.0% of control), ethylacetate fraction (40 ${\mu}g/m{\ell}$, 26.7%), butanol fraction (20 ${\mu}g/m{\ell}$, 87.2%; 40 ${\mu}g/m{\ell}$, 12.5%) of Cremastrae appendiculatae Tuber, water fraction of Persicae Semen (40 ${\mu}g/m{\ell}$, 82.7%), ethylacetate fraction of Hippocampus (40 ${\mu}g/m{\ell}$, 85.3%). The results of gelatin zymogram assay showed that the ingredients of JWCTW decreases the gelatinolytic activity of MMP-9 from ECV304, at the concentration of 10 ${\mu}g/m{\ell}$. In in vitro invasion assay, the ingredients of JWCTW effectively inhibited the invasion of cancer cells as compared with the control (+PMA) groups. In capillary-like tube formation assay, the hexane and ethylacetate fractions of Curcumae Radix, Cremastrae appendiculatae Tuber and Persicae Semen showed the dramatic inhibition effects on tube formation of ECV304 at the concentration of 40 ${\mu}g/m{\ell}$. In ex vivo rat aortic ring assay, the hexane and ethylacetate fractions of Curcumae Radix, Cremastrae appendiculatae Tuber and Persicae Semen showed the inhibition effects on angiogenesis of rat aorta at the concentration of 40 ${\mu}g/m{\ell}$. According to the above research, the anti-angiogenic effects of the ingredients of JWCTW was proved and it suggested that the more effective prescription for anti angiogenesis could be developed.

• Journal of Korean Traditional Oncology
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• v.11 no.1
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• pp.105-118
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• 2006

Two of the essential processes required for metastasis are neoangiogenesis and tumor cell invasion of basement membranes (BM) and extracellular matrix (ECM). Recently, data showed that herbs removing blood stasis has an anti-angiogenic effects. Tonifying vital Qi and eliminating pathogenic factor was a basic modality in Oriental oncology. In this study, we investigated several Qi and Blood tonics for potent angiogenic inhibitors. Methanol extracts of samples inhibited the proliferation of ECV-304 at the concentration of 100 ${\mu}g/m{\ell}$. Zizyphi Fructus, Glycyrrhizae Radix, Angelicae Gigantis Radix decreased the gelatinolytic activity of MMP-9 from ECV-304, at the concentration of 100 ${\mu}g/m{\ell}$ in gelatin zymography. In in vitro invasion assay, herbs inhibited the invasion activity of ECV-304 by 53% of control (Ginseng Radix), 39% (Zizyphi Fructus), 36% (Angelicae Gigantis Radix), 25% (Glycyrrhizae Radix). Ginseng Radix inhibited the capillary-like tube formation of ECV-304 at the concentration of 160 ${\mu}g/m{\ell}$, Angelicae Gigantis Radix and Paeoniae Radix Alba inhibited at the concentration of 320 ${\mu}g/m{\ell}$. These results indicated that Ginseng Radix, Glycyrrhizae Radix, and Angelicae Gigantis Radix could be considered as potent angiogenic inhibitiors.