• The Korean Journal of Pain
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• 제26권3호
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• pp.255-264
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• 2013

Background: We investigated the effects of pre-emptive administration of ketamine and norBNI on pain behavior and the expression of DREAM, c-Fos, and prodynorphin proteins on the ipsilateral side of the rat spinal cord at 2 and 4 hours after formalin injection. Methods: Eighty-four male Sprague Dawley rats were divided into 4 major groups consisting of control rats (C) (n = 12), rats given only formalin injections (F) (n = 24), and rats treated with pre-emptive administration of either ketamine (K+F) (n = 24) or norBNI (N+F) (n = 24). The non-control groups were further divided into subgroups consisting of rats that were sacrificed at 2 and 4 hours (n = 12 for each group) after formalin injection. Pain behavior was recorded for 1 hour. After 2 and 4 hours, the rats were sacrificed and the spinal cords (L4-L5 sections) were removed for immunohistochemistry and Western blot analysis. Results: The pain behavior response was reduced in the K+F group compared to the other groups during the second phase of the formalin pain response. We detected an increase in the nuclear DREAM protein level in the K+F group at 2 and 4 hours and a transient decrease in the N+F group at 2 hours; however, it increased at 4 hours after injection. Fos-like immunoreactivity (FLI) and Prodynorphin-like immunoreactivity (PLI) neurons decreased in the K+F group but increased in the N+F group at 2 hours after injection. While FLI decreased, PLI increased in all groups at 4 hours after injection. Conclusions: We suggest that NMDA and kappa opioid receptors can modulate DREAM protein expression, which can affect pain behavior and protein transcriptional processes at 2 hours and bring about either harmful or protective effects at 4 hours after formalin injection.

• 대한한방종양학회지
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• 제5권1호
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• pp.61-75
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• 1999

To examine the effect of Xuefuzhuyutang on the metastasis of cancer, the following experiments were carried out. Before the main experiments, the cytotoxicity was measured by putting Xuefuzhuyutant sample in HT1080. Then zymography was made to examine the change of gelatinolytic activity. Western blotting was carried out to examine the changes of Fos, Jun, Ets, Erk, md JNK. In vitro invasion assay with transwells coated by collagen and matrigel was carried out. From the above results the following conclusions were obtained. 1. The experimental result about cytotoxicity of Xuefuzhuyutang agaitst HT1080 was a below. The stained cell count after being treated by by Xuefuzhuyutang sample $400{\mu}g/ml$ for 24 hours was 0.9% of total cells, and the stained cell count by Xuefuzhuyutang sample $100{\mu}g/ml$ was 1.5% of total cells. Both were near the level of control group which showed 0.6% stained. 2. The result of collagenase assay was as below. In Xuefuzhuyutang sample $400{\mu}g/ml$, MMP2 was reduced as compared with TPA control group, and the band of MMP-9 induced by TPA disapappeared. In Xuefuzhuyutang samle $800{\mu}g/ml$ both bands of MMP-2 and MMP-9 disapeared. 3. The results of western blots for Jun, Fos, Ets, Erk, JNK were a below. In Xuefuzhuyutang sample $200{\mu}g/ml$, Ets was reduced, and Jun, Fos were increased. 4 The result of invasion assay was as below. The number of cells which migrated across transwell membrane in Xuefuzhuyutang-treated group was less than that of control(+TPA) group. From the above results, it was concluded that Xuefuzhuyutang might inhibit the activity of collagenase not by the MMP-2, MMP-9 promoter but by the other way.

• 대한한방종양학회지
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• 제4권1호
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• pp.131-146
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• 1998

To examine the effect of Shiquandabutang on the metastasis of cancer, the following experiments were made. Before the main experiments, the cytotoxicity was measured by putting Shiquandabutang sample in HT1080. Then zymography was made to examine the change of gelatinolytic activity. And western blotting was carried out to examine the changes of Fos, Jun, Ets, the transcription factors of MMP-2, MMP-9, and Erk, JNK on signal transduction pathway to AP-1. Third, in vitro invasion assay with transwells coated by collagen and matrigel was carried out. From the results of the above the following conclusions were obtained. 1. The experimental result about cytotoxicity of Shiquandabutang against HT1080 was as below. The stained cell count after being treated by Shiquandabutang sample $400{\mu}g/ml$ for 24 hours was 0.9% of total cells, and the stained cell count by Shiquandabutang sample $100{\mu}g/ml$ was 1.5% of total cells. Both were near the level of control group which showed 0.6% stained. 2. The result of collagenase assay was as below. In Shiquandabutang sample $400{\mu}g/ml$, MMP-2 was reduced as compared with TPA control group, and the band of MMP-9 induced by TPA disappeared. In Shiquandabutang sample $800{\mu}g/ml$, both bands of MMP-2 and MMP-9 disappeared. 3. The results of western blots for Jun, Fos, Ets, Erk, JNK were as below. In Shiquandabutang sample $200{\mu}g/ml$, Ets was reduced, and Fos were increased. 4. The result of invasion assay was as below. The number of cells which migrated across transwell membrane in Shiquandabutang-treated group was less than that of +TPA control group. From the above results, it was concluded that Shiquandabutang might control the appearing and acting of collagenase not by the MMP-2, -9 promoter but by other way.

• The Korean Journal of Physiology and Pharmacology
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• 제8권4호
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• pp.213-218
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• 2004

Primary fish-odor syndrome (FOS) is a genetic disorder caused by defective flavin-containing mono-oxygenase 3 gene (FMO3) with deficient N-oxidation of trimethylamine (TMA), causing trimethylaminuria (TMAU). By contrast, secondary FOS can be acquired by decreased FMO activities in patients with chronic liver diseases, but the underlying mechanisms are unknown. In the present study, we examined plasma NOx concentrations and viral DNA contents as well as in vivo FMO activities and their correlations in chronic viral hepatitis (CVH) patients. Plasma concentration of NOx was significantly increased by 2.1 fold $(56.2{\pm}26.5\;vs.\;26.6{\pm}5.4\;{\mu}M,\;p<0.01)$, and it was positively correlated with plasma hepatitis B virus (HBV) DNA contents $(r^2=0.2838,\;p=0.0107)$. Furthermore, the elevated plasma NOx values were inversely and significantly correlated with in vivo FMO activities detected by ranitidine-challenged test $(8.3%\;vs.\;20.0%,\;r^2=0.2109,\;p=\0.0315)$. TMA N-oxidation activities determined in CVH patients without challenge test were also significantly low (73.6% vs. 95.7%, p< 0.05). In conclusion, these results suggested that secondary FOS could be acquired by the endogenously elevated NO in patients with CVH.

• Journal of Acupuncture Research
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• 제18권1호
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• pp.21-28
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• 2001

Objective : The aim of this study was to investigate the effectiveness of stimulation of specific auricular acupuncture points on appetite suppression. Methods : The fasted rats were deprived of food and water for 48 hours. Stimulation of tho inner auricular region of the rat correspond to the human stomach acupuncture point was made for leptin, tryptophan hydroxylase (TPH) and c-fos expression in the brain of the fasted rats by immunohistochemistry. Results : The immunoteactivities of leptin in the periventricular nucleus of the auricular acupunctured group with fasted were significantly higher compared to those of the fasted group. The immunoreactivities of TPH in the median raphe neucleus and reticulotegmental neucleus pons of the auricular acupunctured group with fasted were significantty higher compared to those of the fasted group. And the immunoreactivities of c-fos in the dentate gyrus of the auricular acupunctured group with fasted were significantly lower compared to those from the fasted group. Conclusions : We conclude that the auricular acupuncture altered leptin and TPH-expression in the brain of the fasted rats. The results suggest that auricular acupuncture may inhibit food intake.

• 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.

• 대한의생명과학회지
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• 제11권3호
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• pp.319-326
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• 2005

It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.

• International Journal of Contents
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• 제8권1호
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• pp.74-81
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• 2012

The aim is to investigate the analgesic effect of transcranial direct current stimulation(tDCS) on central neuropathic pain(CNP) in spinal cord contusive rat model. Twenty Sprague-Dawley rats($250{\pm}50$ g, male) were used. Thoracic spinal cord(T10) was contused using New York University(NYU) spinal cord impactor. The animals were randomly assigned to two groups; GroupI: Non-treatment after SCI induction(n=10), GroupII: application of tDCS(0.1 mA, 20 min/time, 2 times/day, 5 days/6week) after SCI induction(n=10). Assess the effect of tDCS using the Basso Beattie Bresnahan(BBB) locomotor rating scales, Touch $test^{TM}$ sensory evaluator(TTSE), Plantar test$^{\circledR}$after contusion at the $2^{nd}$, $3^{rd}$, $4^{th}$, $5^{th}$, $6^{th}$ week and the immunohistochemistric response of c-fos in the thalamus, cerebral cortex after contusion at the $3^{rd}$, $6^{th}$ week after SCI. The scores of BBB scales were significantly different from $3^{rd}$week. TTSE were different significantly over time, but there were no differences at each evaluation times on between-measure time effects. Plantar test were different significantly over time and there were difference at the $4^{th}$, $6^{th}$ week after SCI on between-measure time effects. Also, immunohistochemistric response of c-fos was reduced significantly from $3^{rd}$, $6^{th}$ week after SCI in tDCS group compared with control group in thalamus and cortex. These results identified that tDCS of non-invasive therapeutic method may have beneficial analgesic effect on CNP after SCI with behavioral test and immunohistochemical test.

• Nutritional Sciences
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• 제7권4호
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• pp.196-200
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• 2004

There are increasing evidences that prebiotics can modulate various properties of the immune system. This study was conducted to investigate effects of three kinds of fructans (chicory inulin, chicory inulin oligosaccharide and fructooligosaccharide) and a glucose oligomer(isomaltooligosaccharide) in large bowel mass and innnunoglobulin A (IgA) in rats. Forty five Sprague-Dawley male rats weighing about 1909 were randomly sorted to receive one of the five treatments, which were control diet, control diet+6% isomaltooligosaccharide (IMOS), control diet+6% fructooligosaccharide (FOS), control diet+6% chicory inulin oligosaccharide (CIOS), or control diet + 6% chicory inulin (CI). Rats were pair-fed and received the experimental diets for 5 weeks. Cecal and colonic wall weights were significantly higher in fructan (FOS, CIOS, CI)-fed groups compared with control and IMOS groups, and the length of colon was elevated in FOS and CIOS groups compared with control group. Fecal concentrations of acetic acid and total short-chain fatty acids (SCFAs) were significantly elevated in fructan-fed groups. Plasma and cecal levels and fecal excretion of immunogiobulin A (IgA) in rats were not significantly different among groups. However, fructooligosaccharide tended to increase IgA level in cecum. Cecal IgA level was significantly negatively correlated with pH of cecal content (r=-0.337), positively correlated with acetic acid level (r=0.310). Fecal IgA excretion was positively correlated with total SCFA (r=0.311) and propionic acid (r=0.400) level in feces. These results indicate that fructooligosaccharide and chicory inulin oligosaccharide exerted trophic effects in large bowel wall, increased production of SCFAs and decreased pH, which were conditions positively associated with cecal and colonic IgA secretion.

• Asian Pacific Journal of Cancer Prevention
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• 제15권14호
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• pp.5667-5672
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• 2014

Folate receptor alpha ($FR{\alpha}$) mediates folate uptake by endocytosis, and while folate is essential to DNA methylation and synthesis and may have an important role in proliferating cells. $FR{\alpha}$ is known to be expressed in rapidly proliferating cells, including many cancer cell lines, but there has been no systematic assessment of expression in cervical cancer cell lines. The aim of the present study was to evaluate the effects of $FR{\alpha}$ on proliferation and apoptosis of cervical cells and correlation mechanism. In this study, we investigated the biological function of $FR{\alpha}$ in Hela cells using RNA interference. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK8) assay, while cell cycling and apoptosis were assessed by flow cytometry, mRNA levels by real time-PCR and protein levels of $FR{\alpha}$, c-Fos and c-Jun by Western blotting. The results revealed that $FR{\alpha}$ was highly expressed in Hela cells and its silencing with a small interfering RNA (siRNA) inhibited cell proliferation and induced cell apoptosis, arresting the cell cycle in G0/G1 stages while decreasing the proportion in S and G2/M stages, and suppressed the expression levels of c-Fos and c-Jun. In conclusion, the results of this study indicated that $FR{\alpha}$ down-regulation might be capable of suppressing cervical cancer cell proliferation and promoting apoptosis. It suggested that $FR{\alpha}$ might be a novel therapeutic target for cervical cancer.