• The Korean Journal of Food And Nutrition
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• v.25 no.1
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• pp.181-187
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• 2012

Grifola frondosa has been used as an herbal medicine for the treatment of cancer, diabetes mellitus and high blood pressure. In this study, functional polysaccharide was obtained from Grifola frondosa using four different extraction methods: hot water(HwFP), homogenize(HgFP), acid(AcFP), and alkali(AlFP) extraction methods. The effects of these extracts on KB and HepG2 cell lines were then examined for any anti-cancer activity. Alkaline extraction produced a yield of 0.175% and the total sugar content of the extract was 54.97%. We were able to confirm that the polysaccharide extracts from the mushroom produce an anti-cancer effect. The cytotoxicity of AlFP and AcFP against HepG2 cells were 22.86% and 28.88%, respectively, and the cytotoxicity of AlFP against the KB cell lines was 47.76% at a concentration of 1,000 ${\mu}g/m{\ell}$. Therefore, these results suggest that the optimum method for extracting functional polysaccharides from G. frondosa is the alkali extraction method.

• Journal of Veterinary Clinics
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• v.18 no.4
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• pp.442-447
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• 2001

A 2-year-old 4.0-kg female Shih Tzu with history of hematemesis and melena was referred to Veterinary Medical Teaching Hospital, Seoul national University for further evaluation and treatment. During physical examination, the dog revealed mild depression, dry mucous membrane and abdominal pain. Hematologic values were normal and serum chemical values showed increased serum bile acid (53.47 umol/l, preprandial), fasting serum ammonia concentration (184 g/dl), alanine transferase (98 U/L), alkaline phosphatase (871 U/L) and gamma glutamyl transpeptidase (21 U/L), and decreased blood urea nitrogen (4 mg/dl), total protein (4.1 g/dl) and albumin (1.2 g/dl). Microhepatica was shown in abdominal radiography. During the ultrasound examination, dilated tortuous vein communicating with caudal vena cava ws observed near the stomach. Intraoperative jejunal vein portography was performed during laparotomy to confirm the location and size of shunt vessel. According to history taking, physical examination, hematologic and serum chemical examination and radiographic study, it was diagnosed as single extrahepatic portosystemic shunt. The anomalous vessel (7 mm, o.d.) that enter the caudal vena cava from the left gastric vein, near the level of the diaphragm, was identified. A Ameroid constrictor (5 mm, i.d.) was applied to the shunting vessel near the caudal vena cava. Hematologic and serum chemical values recovered gradually and were revealed normal values 4 months after surgery. Four month after surgery serum bile acids concentrations were 0.56 $\mu$mol/l (preprandial) and 18.45 umol/l (postprandial). Abdominal radiograph showed normal gastric axis and it revealed normal size of the liver. Fine texture and increased echogenecity of liver and enlargement of portal vein were shown in ultrasonography. Single extrahepatic portosystemic shunt might be treated surgically using Ameroid constrictor.

• Biomolecules & Therapeutics
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• v.20 no.1
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• pp.89-95
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• 2012

Poncirus trifoliata fruit (PTF) affects the digestive and cardiovascular systems, and kidney function. The authors studied the effects of ethyl acetate (EtOAc) extract of PTF on the activities of osteoblasts and in an animal model. The main compounds of the EtOAc extract, naringin and poncirin have been confirmed by HPLC and NMR analysis. Effects of osteoblastic differentiation were measured by alkaline phosphatase (ALP) activity, osteopontin (OPN) protein expression and osteoprotegerin (OPG) mRNA expression in MC3T3-E1 cells. Also, osteoclast differentiation was measured by multinucleated cells (MNCs) formation through tartrate resistance acid phosphatase (TRAP)-positive staining. Bone mineral density (BMD) was measured before and after treatment with EtOAc extract of PTF in prednisolone-induced osteoporotic mice. Dexamethasone (DEX) decreased OPN and OPG expression level in MC3T3-E1 cells and ALP activity was decreased by DEX dose-dependently. EtOAc extract of PTF recovered the levels of ALP activity, and the expression of OPN and OPG in MC3T3-E1 cells treated with DEX. In osteoclast differentiation, multinucleated TRAP-positive cell formation was significantly suppressed by the EtOAc extract of PTF. Total body BMD was restored by EtOAc extract of PTF in prednisolone-induced osteoporotic mice. In conclusion, EtOAc extract of PTF recovered DEX-mediated deteriorations in osteoblastic and osteoclastic functions, and increased BMD in glucocorticoid-induced osteoporosis.

• YAKHAK HOEJI
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• v.41 no.5
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• pp.615-621
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• 1997

Dehydropeptidase-I (DHP-I) was solubilized from porcine kidney by treatment with n-butanol and partially purified 19.25 fold by $(NH_4)_2SO_4$ precipitation, DEAE-Sepharose CL-6B ion exchange chromatography and Sephacryl S-300 HR chromatography with an overall yield of 19.16. DHP-I showed its optimal activity at pH 7.5 and 25$^{\circ}C$. Its activity was stable under neutral and alkaline conditions, but was disappeared under acidic condition. And DHP-I was heat-labile and its activity remained at 45$^{\circ}C$ for 3hrs. The enzyme was not inhibited by dicationic ions, while its activity was increased by $Co^{2+}$(1mM) and $Zn^{2+}$ (0.1mM). The enzyme was inhibited by EDTA and N-ethylmaleimide. The relative molecular mass of DHP-I was estimated to be approximately 100kDa by gel filtration chromatography. The $K_m$ value of DHP-I for glycyldehydrophenylalanine (GDHP) was 1.98mM. DWP20418 [(1R, 5S, 6S)-6-[1-(R)-Hydroxyethyl]-1-methyl-2-[(2S, 4S)-2-(piperazinylcarbonyl)-1-(R)-hydroxyethyl)pyrrolidine-4-thio]carbapen-2-em-3-carboxylic acid], compared with meropenem (MEPM), was rather easily hydrolized by DHP-I, while it was four times more resistant than imipenem (IPM) to DHP-I.

• Journal of Veterinary Clinics
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• v.29 no.5
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• pp.400-403
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• 2012

A 10-year-old, intact male, toy poodle was presented with abdominal distension, truncal alopecia, hepatomegaly, and sustained elevation of alkaline phosphatase. Vacuolar hepatopathy and glycogen deposition in hepatocytes were confirmed by liver biopsy and ultrasound-guided fine-needle aspiration with periodic acid-Schiff (PAS) staining of mass lesion respectively. Cortisol and some sex hormones associated with adrenal gland were analyzed at IDEXX Reference Laboratories before and 1 hour after ACTH stimulation. The results of analysis confirmed elevation of some sex hormones including androstenedione, progesterone and 17 hydroxyprogesterone, not cortisol concentration, before and 1 hour after ACTH stimulation. The dog was diagnosed as atypical form of hyperadrenocorticism associated with sex steroids excess. The treatment was initiated with trilostane (0.5 mg/kg, PO, q12hr) that is an adrenal steroid synthesis inhibitor. Trilostane was administered for 8 weeks and the clinical sign including truncal alopecia was improved.

• Journal of Oral Medicine and Pain
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• v.32 no.2
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• pp.129-135
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• 2007

Ecdysteroids are known as insect molting hormone. At the same time, ecdysteroids and plant ecdysteroids (phytoecdysteorids) reveal beneficial effects on mammal. The present study was undertaken to determine the possible cellular mechanism of action of phytoecdysteroids in bone metabolism. The effects on the osteoblasts were determined by measuring cell proliferation, alkaline phosphatase (ALP) activity, and gelatinase activity. The effects on the osteoclasts were investigated by measuring tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation after culturing osteoclast precursors. Phytoecdysteroid treatment showed a increase in ALP activity of osteoblasts. Phytoecdysteroid increased the activity of gelatinase. In addition, phytoecdysteroid decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL) in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, phytoecdysteroid may be a regulatory protein within the bone marrow microenvironment.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.2
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• pp.19-35
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• 2020

Objectives The aim of this study is to evaluate the fracture healing effect of Jinmu-tang (JM) on femur fractured rats. Methods Rats were randomly divided into 5 groups (normal, control, positive control, JM extract with low concentration and JM extract with high concentration). All group except normal group went through both femur fracture. Normal and control group received no treatment at all. Positive control group were medicated with tramadol (20 mg/kg) once a day for 14 days. Experimental group was orally medicated with JM extract (10 mg/kg for low concentration, 50 mg/kg for high concentration) once a day for 14 days. In order to investigate fracture healing process, plasma and serum were obtained. Also, micro-computed tomography was conducted to see the frature site visually. Immunohistochemistry for transforming growth factor-β1, Ki67, alkaline phosphatase, runt-related transcription factor 2, receptor activator of nuclear factor kappa-β, tartrate resistant acid phosphatase was conducted to observe bone healing progress after 14 days since fracture occured. Aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen and creatinine levels were measured in plasma, for hepatotoxicity and nephrotoxicity of JM extract. Osteocalcin was measured to observe activity of osteoblast. Results Through Micro-CT, more fracture healing was observed on both experimental group than control and positive control group. Through Hematoxylin & Eosin and safranin O staining showed bone cell proliferation and bone formation in the experimental group. RANK was significantly increased in the experimental groups. JM with high concentration showed statistically significant of TGF-β and Osteocalcin. NO, TRAP and ALP were not significantly changed. Liver toxicity was not significantly observed. Creatinine significantly increased in both experimental groups after 28 days. Conclusions As described above, JM extract showed anti-inflammatory effect, promoted fracture healing by stimulating the bone regeneration factor, and showed little hepatotoxicity and nephrotoxicity. In conclusion, JM extract can promote fracture healing and it can be used clinically to patients with fracture.

• Molecules and Cells
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• v.43 no.1
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• pp.34-47
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• 2020

The circadian clock regulates various physiological processes, including bone metabolism. The nuclear receptors Reverbs, comprising Rev-erbα and Rev-erbβ, play a key role as transcriptional regulators of the circadian clock. In this study, we demonstrate that Rev-erbs negatively regulate differentiation of osteoclasts and osteoblasts. The knockdown of Rev-erbα in osteoclast precursor cells enhanced receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast formation, as well as expression of nuclear factor of activated T cells 1 (NFATc1), osteoclast-associated receptor (OSCAR), and tartrate-resistant acid phosphatase (TRAP). The overexpression of Rev-erbα leads to attenuation of the NFATc1 expression via inhibition of recruitment of c-Fos to the NFATc1 promoter. The overexpression of Rev-erbα in osteoblast precursors attenuated the expression of osteoblast marker genes including Runx2, alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OC). Rev-erbα interfered with the recruitment of Runx2 to the promoter region of the target genes. Conversely, knockdown of Rev-erbα in the osteoblast precursors enhanced the osteoblast differentiation and function. In addition, Rev-erbα negatively regulated osteoclast and osteoblast differentiation by suppressing the p38 MAPK pathway. Furthermore, intraperitoneal administration of GSK4112, a Rev-erb agonist, protects RANKL-induced bone loss via inhibition of osteoclast differentiation in vivo. Taken together, our results demonstrate a molecular mechanism of Rev-erbs in the bone remodeling, and provide a molecular basis for a potential therapeutic target for treatment of bone disease characterized by excessive bone resorption.

• Journal of Oral Medicine and Pain
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• v.31 no.1
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• pp.17-25
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• 2006

The present study was undertaken to determine the possible cellular mechanism of action of angiopoietin-2 in bone metabolism. The effects on the osteoblasts were determined by measuring 1) cell viability, 2) alkaline phosphatase (ALP) activity, 3) gelatinase activity, and 4) nitric oxide production. The effects on the osteoclasts were investigated by measuring 1) tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation, and 2) resorption areas after culturing osteoclast precursors. Angiopoietin-2 treatment showed a significant increase in both the viability and ALP activity of osteoblasts. Angiopoietin-2 increased the activity of gelatinase and nitric oxide production. In addition, angiopoietin-2 decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL), and inhibited osteoclastic activity in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, angiopoietin-2 may be a regulatory protein within the bone marrow microenvironment.

• Advances in nano research
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• v.4 no.1
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• pp.1-14
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• 2016

Superparamagnetic iron oxide nanoparticles (Nps), composed of magnetite, $Fe_3O_4$, or maghemite, ${\gamma}-Fe_2O_3$, core and biocompatible polymer shell, such as dextran or chitozan, have recently found wide applications in magnetic resonance imaging, contrast enhancement and hyperthermia therapy. For different diagnostic and therapeutic applications, current attempt is focusing on the synthesis and biomedical applications of various ferrite Nps, such as $CoFe_2O_4$ and $MnFe_2O_4$, differing from iron oxide Nps in charge, surface chemistry and magnetic properties. This study is focused on the synthesis of manganese ferrite, $MnFe_2O_4$, Nps by most commonly used chemical way pursuing better control of their size, purity and magnetic properties. Co-precipitation syntheses were performed using aqueous alkaline solutions of Mn(II) and Fe(III) salts and NaOH within a wide pH range using various hydrothermal treatment regimes. Different additives, such as citric acid, cysteine, glicine, polyetylene glycol, triethanolamine, chitosan, etc., were tested on purpose to obtain good yield of pure phase and monodispersed Nps with average size of ${\leq}20nm$. Transmission electron microscopy (TEM), X-ray diffraction, energy dispersive X-ray spectroscopy (EDX), $M\ddot{o}ssbauer$ spectroscopy down to cryogenic temperatures, magnetic measurements and inductively coupled plasma mass spectrometry were employed in this study.