• Journal of Veterinary Clinics
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• v.28 no.6
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• pp.549-554
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• 2011

FK506 is a widespread immunosuppressive drug after liver transplantion in patients with advanced-stage hepatocellular carcinoma. Dexamethasone is frequently used as co-treatment in cytotoxic cancer therapy, e.g. to prevent nausea, to protect normal tissue or for other reasons. Our aim was to investigate antitumor effects of FK506 in Hep3B cells, one of differentiated human hepatocellular carcinoma cell lines and inhibitory effects of dexamethsone on FK506- induced antitumor effects. Cell injury was evaluated by biochemical assays as cell viability, lactate dehydrogenase (LDH) and reactive oxygen species (ROS) in Hep3B cells. Intracellular calcium concentration ([$Ca^{2+}$]i) and the level of activation of the c-Jun-N-terminal kinase (JNK) and the Bax protein in cultured Hep3B cells was measured. Exposure of 0.1 ${\mu}M$ FK506 to Hep3B cells led to cell death accompanied by a decrease in cell viability and an increase in LDH, ROS and [$Ca^{2+}$]i. FK506 induced an increase in activity of Bax and JNK protein but inhibited the activity of Bcl-2 protein. Treatment of dexamethsone, per se, had no effects on cell viability, LDH and ROS. However, co-treatment of FK506 and dexamethasone diminished the FK506-induced LDH release, ROS generation and JNK activation. These results demonstrate that FK506 has antitumor effect in Hep3B cells but the combination of FK506 and dexamethasone antagonizes the FK506-induced antitumor effects.

• Restorative Dentistry and Endodontics
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• v.21 no.1
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• pp.321-338
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• 1996

The purpose of this study was to evaluate the effect of moistening and air-drying of acid conditioned dentin and enamel on the marginal microleakage. In this study, Class V cavity were prepared on both buccal and lingual surface of sixty extracted human premolars with cementum margin. These specimens were randomly devided into three groups and three dentin adhesives(Scotchbond Multi-Purpose, All bond 2, Prisma Universal Bond 3) were applied to each group. The specimens in each group were subdevided into four groups (Wet/primed, Dry/primed, Wet/not primed, Dry/not primed) and the etched dentin and enamel surface were treated these four surface treatments prior to the placement of a bonding agent or adhesive. Wet/primed group was simply blot-dried with a damp facial tissue before primer placement ; Dry/primed group was air dried for 30 seconds before the placement of a primer ; Wet/not primed group and Dry/not primed group were not primed after blot dried and air dried for 30 seconds each group. The bonding agent and composite resin were applied for each group. All specimens were exposed to 500 cycle of thermal stress. Specimens were placed in a silver nitrate solution and then sectioned buccolingually through the center of the restoration. The dye penetrations of the specimens were observed with a stereo microscope. The statistical test were applied to the results using a one way analysis variance (ANOVA) and Duncan's multiple range test. The aspects of silver ion penetration into the resin/dentin interface were examined under scanning electron microscopy. The results were as follows. 1. In all groups, the enamel margin showed significantly lower leakage value than the cementum margin (p<0.05). 2. Regardless of various surface treatment and dentin adhesives, there was no significant difference at the enamel margins (p>0.05). 3. At the dentin margins, the leakage values of Dry/not primed group showed significantly higher than that of the other groups (p<0.05). The leakage values of Wet/primed group showed significantly lower than that of the other groups, but, there was no significant difference between Wet group and Dry group. 4. There was no significantly difference between the dentin adhesives regarding the surface treatments in all groups(p>0.05). 5. On the backscatterd scanning electron microscopy observation, the penetration of the silver ion occured at the bonding resin/dentin interface. In the Wet/primed group, resindentin hybrid zone and resin penetration into the dentin was observed. The resin tags were compactively formed to a thickness of $3\sim4{\mu}m$ at the upper part of dentinal tubules. In the Dry/primed group, the thickness of the hybrid zone and the diameter, depth of the resin tags diminished. In the Non-primed groups, the hybrid zone was not identified and few resin tag was observed. There was the gap formation in the resin/dentin interface.

• Journal of Acupuncture Research
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• v.27 no.6
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• pp.31-42
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• 2010

Objectives : The aim of this study was to investigate the anti-obesity potential and mechanisms of action of Rhizoma Atractylodis(RA) herbal acupuncture in high fat diet- induced obese ICR mice. Methods : Sample solutions for herbal acupuncture were prepared from the Rhizoma Atractylodis water extract powder at concentration of 150mg/kg and 300mg/kg with distilled water. Five week-old ICR mice acclimatized to the laboratory environment for 1 week were allocated into four groups: regular diet group (RD), high fat diet group(HFD), groups fed HFD with 150mg/kg RA herbal acupuncture treatment (RAE 150) and with 300mg/kg RA herbal acupuncture treatment(RAE 300). Herbal acupuncture groups were injected with either 150mg/kg or 300mg/kg of Rhizoma Atractylodis(RA) subcutaneously onto both Sinsu($BL_{23}$) alternately on the same time everyday for 30days. Body weight, gross appearance of epididymal fat area, blood glucose, insulin, insulin resistance(HOMA-IR), non-esterified fatty acid, cholesterol, triglyceride, AST, ALT, histological analysis of white adipose tissue, gene expression responsible for adipocyte differentiation and AMPK activation were analyzed. Results : RA herbal acupuncture inhibited the development of weight gain, hyperglycemia, hyperinsulinemia, hyperlipidemia, increases of AST and ALT, and the enlargement of fat cell size induced by HFD. Also, RA herbal acupuncture inhibited the expression of PPAR-${\gamma}$, C/$EBP{\alpha}$, aP2, LPL, FAS, SCD-1 and enhanced the activation of AMP-activated protein kinase. Conclusions : The results of this study demonstrate that RA herbal acupuncture can exert the anti-obesity effect and it is partially mediated by activation of AMPK and inhibition of the gene expressions responsible for adipocyte differentiation. Further studies will be required to ascertain the nti-obesity effect and mechanisms of action of RA herbal acupuncture in animal models and human for aclinical application.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.4
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• pp.314-320
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• 2001

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

• Nutrition Research and Practice
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• v.11 no.3
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• pp.190-197
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• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• Development and Reproduction
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• v.15 no.2
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• pp.113-120
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• 2011

BCL-2 family essential proteins to play a pivotal role to perform in apoptosis signaling pathways and essential proteins for the regulation of cell death. BCL2L10 protein is a member of BCL-2 family and it regulates both anti-apoptotic and pro-apoptotic function of specific tissue or cell line. BCL2L10 of function and expression is not reported in ovary cell lines. In this study we reported that BCL2L10 were significant expression of KGN cell line. Ectopic expression of BCL2L10 induced cell death, and its cells killing effect was blocked by pan-caspase inhibitor of the Z-VAD-fmk. Ectopic expression of BCL2L10 protein led to the activation of caspase 9 and caspase 3, suggesting apoptotic cell death, and confocal microscopic analyses showed that BCL2L10 was partially localized in mitochondria. Thus, we provide a novel function of BCL2L10 in KGN cells, which was involved in the intrinsic cell death pathway.

• The Korean Journal of Pharmacology
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• v.17 no.2
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• pp.37-45
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• 1981

The importance of thyroid hormones for the survival of rats in the cold is along-established fact. Hypothyroid animals are unable to survive in a cold environment. It was also reported that acute exposure of rats, guinea pigs and rabbits to cold produced an increased secretion of TSH and thereby thyroid hormone secretion within 10 to 30 min, but this increase of thyroid activity disappeared quite rapidly during warming. However, in human study no significant difference was found in the concentration of $T_4$, TSH and cortisol between summer and winter. But plasma $T_3$ concentration was increased significantly in winter in 56 adult men. On the other hand, it has been also known that catecholamines are important in the maintenance of body temperature of rat exposured to cold. Abundant evidences suggest that the sympathetic nervous system is involved in the activation of nonshivering thermogenesis and that thyroid hormone metabolism and secretion are influenced by catecholamines and consequently by the activity of the sympatheticadrenal system. Many of the metabolic effects of catecholamines are associated with an increase in the level of cAMP mediated through activation of adenylate cyclase which converts ATP to cAMP. Other studies have shown that thyroid hormones affect the amount of adenylate cyclase present in the adipose tissue. On the other hand. it was also reported that a particulate cAMP phosphodiesterase activity in fat cells was modulated by the action of thyroid hormones. The objective of the present study was to determine the interaction between thyroid activity and cyclic nucleotides during acute exposure to cold. Albino rats weighing around 200 g were used as the experimental animal. The room temperature group was kept at $25^{\circ}C$ and the cold-exposured group was kept at $4^{\circ}C$ for 1 week or 2 weeks. Each group was subdivided into three subgroups; control, KI, and MTU group. At the end of experiment the animals were etherized and blood was taken from abdominal aorta for $T_4,\;T_3$ and cyclic nucleotides. The determinations of $T_3,\;T_4$ and cyclic nucleotides were carried out with a radioimmunoassay(RIA) method. The results were summerized as followings. 1) A significant increase of thyroid weight was observed in rats exposured to cold for 2 weeks. Furthermore, in rats administered MTU while to exposure to cold the thyroid weight was also increased significantly. 2) After 2 weeks $T_3$ concentration in the plasma of cold-exposured rats was significantly increased in KI group and MTU group as well as in control group. On the contrary, after 2 weeks of cold exposure $T_4$ level was decreased in control group. 3) In the case of cyclic nucleotides, plasma cAMP was increased in the control group after 1 or 2 weeks of cold exposure. However, cAMP level in plasma was rather significantly decreased in KI group and MTU group as well as in control group.

• Journal of Embryo Transfer
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• v.31 no.4
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• pp.335-347
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• 2016

Multiple interferon tau (IFNT) genes exist in bovine. An antiluteolytic substance secreted by the bovine conceptus and primarily responsible for maternal recognition of pregnancy is bovine trophoblast protein 1 (bIFNT1), a new type I interferon tau (IFNT) genes. The objectives of this research were to investigate whether multiple, distinct gene encode bIFNT1 and other type I bIFNT gene in the bovine genome and to examine expression of bIFNT1 and other bIFNTc1 mRNAs during conceptus development. These transcrips could be regulated through caudal-related homeobox-2 (CDX2) and ETS2 and/or AP1 (JUN) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. The presence of mRNAs encoded by bIFNT1 and type I bIFNTc1 genes were examined quantitatively via reverse transcription-polymerase chain reaction (RT-PCR) analysis of total cellular RNA (tcRNA) extracted from on day 17, 20 and 22 bovine conceptuses. The expression level of bIFNT1 was higher on day 17 transcripts were gradually weakly detectable on day 20 and 22. However, the other bIFNTc1 gene examined transcripts was highly expressed on day 20 and transcripts were weakly detectable on day 17 and 22 bovine conceptuses. Furthermore, human choriocarcinoma JEG3 was co-transfected with an -1kb-bIFNT1/c1-Luc constructs and several transcription factor expression plasmids. Compared to each -1kb-bIFNT1/c1-Luc increased when this constructs were co-transfected with, ETS2, AP1(JUN), CREBBP and/or CDX2. Also, bIFNTc1 gene was had very effect on activity by alone ETS2, and AP1 (JUN) expression factors in choriocarcinoma JEG3 cell. However, bIFNT1 gene expression of the upstream region was not identified. We demonstrated that the activities of bIFN genes are regulated by differential, tissue-specific and developmental competence during pregnancy.

• Reproductive and Developmental Biology
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• v.33 no.2
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• pp.107-111
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• 2009

The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

• Journal of Nutrition and Health
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• v.53 no.1
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• pp.1-12
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• 2020

Purpose: Diabetic nephropathy is one of the most important diabetic complications prompted by chronic hyperglycemia, characterized by glomerulosclerosis, tubular fibrosis, and it eventually causes kidney failure. Nobiletin is a polymethoxyflavone present in tangerine and other citrus peels, and has anti-cancer and anti-inflammatory effects. This study investigated the effects of nobiletin on glomerular fibrosis through inhibition of the transforming growth factor (TGF)-β1-Src-caveolin-1 pathway. Methods: Human renal mesangial cells (HRMC) were incubated in media containing 33 mM glucose with or without 1-20 uM nobiletin for 3 day. The cellular expression levels of fibrogenic collagen IV, fibronectin, connective tissue growth factor (CTGF), TGF-β1, Src and caveolin-1 were all examined. In addition, TGF-β1, Src and caveolin-1 proteins were screened to reveal the relationship among TGF-β1-Src-caveolin-1 signaling in glomerular fibrosis. Results: High glucose promoted the production of collagen IV, fibronectin and CTGF in HRMC, which was inhibited in a dose dependent manner by 1-20 uM nobiletin. The Western blot data showed that high glucose elevated the expression of TGF-β1, Src, caveolin-1 and Rho GTPase. When nobiletin was treated to the HRMC exposed to high glucose, the expression of TGF-β1-Src-caveolin-1 was dampened. Finally, TGF-β1-Src-caveolin-1 signaling pathway was activated in high glucose-exposed HRMC, and such activation was encumbered by nobiletin. Conclusion: These result demonstrated that nobiletin blunted high glucose-induced extracellular matrix accumulation via inhibition of the TGF-β1-Src-caveolin-1 related intracellular signaling pathway. Nobiletin may be a potent renoprotective agent to counteract diabetes-associated glomerular fibrosis that leads to kidney failure.