• Journal of Life Science
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• v.22 no.6
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• pp.760-771
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• 2012

Marine triterpene glycosides are physiologically active natural compounds isolated from sea cucumbers(holothurians). It was demonstrated that they have a wide range of biological activities, including antifungal, cytotoxic, and antitumor effects. A previous study showed that stichoposide C (STC) isolated from Thelenota anax induces apoptosis through generation of ceramide by activation of acid sphingomyelinase (SMase) and neutral SMase in human leukemia cells. In this study, we investigated whether STD, a structural analog of STC, can induce apoptosis and examined the molecular mechanisms for its activity. It was found that STC and STD induce apoptosis in a dose- and time-dependent manner and lead to the activation of caspase-8, mitochondrial damage, activation of caspase-9, and activation of caspase-3 in K562 and HL-60 cells. STC activates acid SMase and neutral SMase, which results in the generation of ceramide. Specific inhibition of acid SMase or neutral SMase partially blocked STC-induced apoptosis, but not STD-induced apoptosis. In contrast, STD generates ceramide through the activation of ceramide synthase. Specific inhibition of ceramide synthase partially blocked STD-induced apoptosis, but not STC-induced apoptosis. Moreover, STC and STD markedly reduced tumor growth of HL-60 xenograft tumors and increased ceramide generation in vivo. These results indicate that STC and STD can induce apoptosis and have antitumor activity through the different molecular mechanisms, because they have a different sugar residue attached to aglycones. Thus, these results suggest that their actions are affected by a sugar residue attached to aglycones and they can be used as anticancer agents in the treatment of leukemia.

• Journal of Life Science
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• v.21 no.2
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• pp.183-193
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• 2011

Migration and differentiation of mesenchymal stem cells are crucial for tissue regeneration in response to injury. Sphingosine-1-phosphate (S1P) is a bioactive lipid that regulates a variety of biological processes, including proliferation, survival, differentiation and motility. In the present study, we determined the role of S1P in migration and differentiation of human bone marrow-derived mesenchymal stem cells (BMSCs). S1P stimulated migration of BMSCs in a dose- and time-dependent manner, and pre-incubation of the cells with pertussis toxin completely abrogated S1P-induced migration, suggesting involvement of Gi-coupled receptors in S1P-induced cell migration. S1P elicited elevation of intracellular concentration of $Ca^{2+}$ ($[Ca^{2+}]_i$) and pretreatment with VPC23019, an antagonist of $S1P_1/S1P_3$, blocked S1P-induced migration and increase of $[Ca^{2+}]_i$. Small interfering RNA-mediated knockdown of endogenous $S1P_1$ attenuated S1P-induced migration of BMSCs. Furthermore, S1P treatment induced expression of $\alpha$-smooth muscle actin ($\alpha$-SMA), a smooth muscle marker, and pretreatment with VPC23019 abrogated S1P-induced $\alpha$-SMA expression. S1P induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), and pretreatment of cells with SB202190, an inhibitor of p38 MAPK, or adenoviral overexpression of a dominant-negative mutant of the p38 MAPK blocked S1P-induced cell migration and $\alpha$-SMA expression. Taken together, these results suggest that S1P stimulates migration and smooth muscle differentiation of BMSCs through an $S1P_1$-p38 MAPK-dependent mechanism.

• Journal of the Korean Orthopaedic Association
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• v.53 no.6
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• pp.513-521
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• 2018

Purpose: The purpose of this study was to assess the effectiveness and complications of an ultrasound-guided axillary brachial plexus block performed by orthopedic surgeons. Materials and Methods: From March to May 2017, an ultrasound-guided axillary brachial plexus block was performed on a total of 103 cases of surgery. A VF13-5 transducer from Siemens Acuson X300 was used. The surgical site was included in the range of the anatomic sensory distribution of the blocked nerve, except for the case where an operation time of more than 2 hours was expected due to multiple injuries and the operation of the upper arm. The procedure was performed by 2 orthopedic surgeons in the same method using 50 ml of solution (20 ml of lidocaine HCl in 2%, 20 ml of ropivacaine in 0.75%, 10 ml of normal saline in 0.9%). The success rate of anesthesia induction during surgery, anesthetic induction time, anatomical range of operation, duration of postoperative analgesia and complications were investigated. Results: The results from the 2 practices were similar. The anesthesia was successful in 100 out of 103 patients (97.1%). In these patients, the average needling time was 5.5 minutes (2.5-13.2 minutes), the average induction time to complete anesthesia was 18.4 minutes (5-40 minutes), and the average duration of postoperative analgesia was 402.8 minutes (141-540 minutes). The post-anesthesia immediate complications were dizziness in 1 case, nausea and vomiting in 4 cases, and peri-oral numbness in 2 cases, but surgery was performed without problems. All these 7 cases with complications recovered on the same day. A total of 3 cases failed with anesthesia, and they were treated by an injection with local anesthesia in the operation room in 2 cases and switched to general anesthesia in 1 case. Conclusion: An ultrasound-guided axillary brachial plexus block, which was performed by orthopedic surgeons allows anesthesia in a brief period and the high success rates of anesthesia for certain surgeries of the elbow and surgeries on forearm, wrist and hand. Therefore, it can reduce the waiting time to the operating room. This technique is a relatively safe procedure and dose selective anesthesia is possible.

• Journal of Korean Society of Environmental Engineers
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• v.36 no.8
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• pp.554-560
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• 2014

To investigate the applicability of montmorillonite to capping material for the remediation of contaminated marine sediment, adsorption characteristics of $PO{_4}{^{3-}}$ onto montmorillonite were studied in a batch system with respect to changes in contact time, initial concentration, pH, adsorbent dose amount, competing anions, adsorbent mixture, and seawater. Sorption equilibrium reached in 1 h at 50 mg/L but 3 h was required to reach sorption equilibrium at 300 mg/L. Freundlich model was more suitable to describe equilibrium sorption data than Langmuir model. The $PO{_4}{^{3-}}$ adsorption decreased as pH increased, due to the $PO{_4}{^{3-}}$ competition for favorable adsorption site with OH- at higher pH. The presence of anions such as nitrate, sulfate, and bicarbonate had no significant effect on the $PO{_4}{^{3-}}$ adsorption onto the montmorillonite. The use of the montmorillonite alone was more effective for the removal of the $PO{_4}{^{3-}}$ than mixing the montmorillonite with red mud and steel slag. The $PO{_4}{^{3-}}$ adsorption capacity of the montmorillonite was higher in seawater than deionized water, resulting from the presence of calcium ion in seawater. The water tank elution experiments showed that montmorillonite capping blocked well the elution of $PO{_4}{^{3-}}$, which was not measured up to 14 days. It was concluded that the montmirillonite has a potential capping material for the removal of the $PO{_4}{^{3-}}$ from the aqueous solutions.

• The Korean Journal of Nuclear Medicine
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• v.7 no.2
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• pp.1-12
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• 1973

The most commonly used methods for determining thyroxine binding globulin(TBG) concentration as the total thyroxine-binding capacity utilize electrophoretic seperation of serum. Although technically simple, the electrophoretic method is time consuming and is limited in the number of samples which can be run in a single assay. The author presented a single $T_4$ load ion exchange resin method as an approach to simplify the technique as with clinical practicability and results were analyzed. For construction of the standard curves, serum mixtures were diluted with barbital buffer.which effectively blocked $T_4$-binding to TBPA. For each serum dilution, a constant amount of $T_4-^{125}I$ and increments of unlabelled $T_4$ were added. After incubation in water bath, resin beads were dispensed to the samples which binded all $T_4$ not bound to TBG. The radioactivity in the supernatant was counted in the gamma scintillation counter. Each standard curve was plotted from the percent counts in the supernatant and total $T_4$ in each tube. Unknown samples were diluted to 1:40 and ran at a single $T_4$ loading concentration, and the TBG capacity of the samples was able to be read on the standard isobars. The following results were obtained. 1) Mean and standard deviation for TBG capacity in normal population was $28.6{\pm}5.09{\mu}g\;T_4/100ml$. 2) $24.9{\pm}3.87{\mu}g\;T_4/100ml$ in hyperthyroidism showed low TBG capacity comparing to normal population.(p<0.025) 3) $31.0{\pm}2.40{\mu}g\;T_4/100ml$ in hypothyroidism showed high TBG capacity tendency comparing to normal population. 4) Reversed correlationship existed between TBG capacity and $T_3$ resin uptake(r=-0.624), TBG capacity and serum $T_4$ value (r=-0.859), and TBG capacity and free thyroxine index(r=-0.623). The author assumes that this method of assay is considerably simpler in instrumentation and technique than any other assays traditionally being used, and seems to be more practical for routine clinical laboratory use.

• Biomolecules & Therapeutics
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• v.16 no.2
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• pp.147-160
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• 2008

The present study was designed to examine effects of polyphenolic compounds isolated from red wine (PCRW) on the release of catecholamines (CA) from the isolated perfused model of the rat adrenal medulla, and to clarify its mechanism of action. PCRW (20${\sim}$180 ${\mu}$g/mL), given into an adrenal vein for 90 min, caused inhibition of the CA secretory responses evoked by ACh (5.32 mM), high $K^+$ (a direct membrane-depolarizer, 56 mM), DMPP (a selective neuronal nicotinic $N_N$ receptor agonist, 100 ${\mu}$M) and McN-A-343 (a selective muscarinic $M_1$ receptor agonist, 100 ${\mu}$M) in dose- and time-dependent fashion. PCRW itself did not affect basal CA secretion (data not shown). Following the perfusion of PCRW (60 ${\mu}$g/mL), the secretory responses of CA evoked by Bay-K-8644 (a L-type dihydropyridine $Ca^{2+}$ channel activator, 10 ${\mu}$M), cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, 10 ${\mu}$M) and veratridine (an activator of voltage-dependent $Na^+$ channels, 10 ${\mu}$M) were also markedly blocked, respectively. Interestingly, in the simultaneous presence of PCRW (60 ${\mu}$g/mL) and L-NAME (a selective inhibitor of NO synthase, 30 ${\mu}$M), the inhibitory responses of PCRW on the CA secretion evoked by ACh, high $K^+$, DMPP, McN-A-343, Bay-K-8644 and cyclpiazonic acid were recovered to considerable level of the corresponding control release compared with those effects of PCRW-treatment alone. Practically, the amount of NO released from adrenal medulla after loading of PCRW (180 ${\mu}$g/mL) was significantly increased in comparison to the corresponding basal released level. Collectively, these results obtained here demonstrate that PCRW inhibits the CA secretory responses evoked by stimulation of cholinergic (both muscarinic and nicotinic) receptors as well as by direct membrane-depolarization from the isolated perfused adrenal gland of the normotensive rats. It seems that this inhibitory effect of PCRW is mediated by blocking the influx of both ions through $Na^+$ and $Ca^+{2$} channels into the rat adrenomedullary chromaffin cells as well as by inhibiting the release of $Ca^{2+}$ from the cytoplasmic calcium store, which are due at least partly to the increased NO production through the activation of nitric oxide synthase. Based on these data, it is also thought that PCRW may be beneficial to prevent or alleviate the cardiovascular diseases, such as hypertension and angina pectoris.

• Journal of Life Science
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• v.30 no.9
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• pp.772-782
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• 2020

Skeletal muscle differentiation or myogenesis is important to maintain muscle mass and metabolic homeostasis. Muscle-specific microRNAs (miRNAs) are known to play a critical role in skeletal myogenic differentiation. In this study, we examined the expression profiling of miRNAs during myogenic differentiation in rat L6 myoblasts using rat miRNA microarrays. We identified the upregulated expression of miR-128 as well as several well-known myogenic miRNAs, including miR-1, miR-133b, and miR-206. We additionally confirmed the increased expression of miR-128 observed on microarray through quantitative real-time PCR (qRT-PCR), which showed similarly upregulated expression of both primary miR-128 and mature miR-128, consistent with the microarray findings. Furthermore, transfection of miR-128 into rat L6 myoblasts induced gene expression of myogenic markers such as muscle creatine kinase (MCK), myogenin, and myosin heavy chain (MHC). Protein expression of MHC was increased as well. Inhibition of miR-128 by inhibitory peptide nucleic acids (PNAs) blocked the expression of those myogenic markers. In addition, the transfection of miR-128 into rat L6 myoblasts enhanced the phosphorylation of Erk and Akt proteins stimulated by insulin, while simultaneously reversing the inhibited phosphorylation of Erk and Akt due to insulin resistance. These findings suggest that miR-128 may play important roles in myogenic differentiation and insulin signaling.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.4
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• pp.231-238
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• 2005

Neuronal apoptotic events, which result in cell death, are occurred in hypoxic/ischemic conditions. Estradiol is a female sex hormone with steroid structure known to provide neuroprotection through multiple mechanisms in the central nervous system. This study was aimed to investigate the signal transduction pathway of $CoCl_2$-induced neuronal cell death and the inhibitory effects of estradiol. Administration of $CoCl_2$ decreased cell viability in both a dose- and time-dependent manner in PC12 cells. $CoCl_2$-induced cell death produced genomic DNA fragmentation and morphologic changes such as cell shrinkage and condensed nuclei. It was found that $CoCl_2$-treated cells increased the reactive oxygen species (ROS) as well as caspase-8, -9 and -3 activities. However, pretreatment with estradiol before exposure to $CoCl_2$ prevented the reduction in cell viability reduction and attenuated DNA fragmentation and morphologic changes caused by $CoCl_2$. Furthermore, the $CoCl_2$-induced increases of ROS levels and caspases activities were attenuated by estradiol. Gene expression analysis revealed that estradiol blocked the underexpression of the Bcl-2 and ameliorated the increase in the release of cytochrome c from mitochondria into cytoplasm and Fas-ligand (Fas-L) upregulated by $CoCl_2$. These results suggest that $CoCl_2$ induce apoptosis in PC12 cells through both mitochondria- and death receptor-mediated cell death pathway. Estradiol was found to have a neuroprotective effect against $CoCl_2$-induced apoptosis through the inhibition of ROS production and by modulating apoptotic effectors associated with the mitochondria- and death-dependent pathway in PC12 cells.

• The Korea Journal of Herbology
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• v.22 no.3
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• pp.27-37
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• 2007

Objectives: Hepatocellular carcinoma is the most common primary malignant tumor of the liver worldwide. In-Jin-Ho-Tang(IJHT) has been used as a traditional Chinese herbal medicine since ancient time. and today it is widely applied as a medication for jaundice which is associated with inflammation in liver. In this study, I investigated whether methanol extract of IJHT induced HepG2 cancer cell death. Methods: Cytotoxic activity of IJHT on HepG2 cells was using XTT assay. Apoptosis induction by Ros A in HCT116 cells was verified by the induction of cleavage of poly ADP-ribose polymerase (PARP). and activation of caspase-3, -8 and -9. The release of cytochrome c from mitochondria to cytosol. the level of Bcl-2 and Bax and the expression of p53 and p21 were examined by western blotting analysis. Furthermore, MAPKs activation was analyzed by western blotting analysis. Results: IJHT induced apoptosis in HepG2 cells. And treatment of IJHT resulted in the release of cytochrome c into cytosol, decreased anti-apoptotic Bcl-2, and increased pri-apoptotic Bax expression. IJHT markedly inactivated extracellular signal-regulated kinase (ERK1/2), and activated p38 mitogen-activated protein (MAP) kinase. Sodium orthovanadate (SOV), a phosphatase inhibitor, to reverse IJHT-induced ERK1/2 inactivation and SB203580, a specific p38 MAP Kinase inhibitor efficiently blocked apoptosis of HepG2. Thus, IJHT induces apoptosis in HepG2 cells via MAP kinase modulation. Conclusion: These results indicated that IJHT has some potential for use as an anti-cancer agent.

• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.333-350
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• 1999

The modulation of leukocyte cell surface adhesion molecules may influence the development of cellular events that determine the course of the inflammatory process. Direct interaction between activated T cells and monocytes resulted in a large production of $IL-1{\beta}$ by monocytes. In this reactions, adhesion molecules play an important part, yet the role of them in Tmonocytes interaction remain unclear. This study was undertaken in an effort to elucidate, 1) the influence of 1.25(OH)$_2D_3-induced$ differentiation on the monocyte responsiveness to direct contact with T lymphocytes, and 2) the role of adhesion molecules on the T-monocyte direct interaction. Initially, I observed that direct contact of monocyte cell line THP-1 with stimulated fixed T cell line HuT78 markedly induces IL-1${\beta}$ production by THP-1. $IL-1{\beta}$ production was higher when THP-1 had been previously exposed to 1.25(OH)$_2D_3$ as compared to control, with ${\alpha}$- 1.25(OH)$_2D_3$ dose-dependent and exposure time-dependent manner. It was shown that 1.25(OH)$_2D_3$ also increased the expression of ${\beta}_2$ integrin adhesion receptor Mac-1(CD11b/CD18) dose- and timedependently, but did not increase the expression of human leukocyte antigen- D(HLA-D) and intercellular adhesion molecule-1(ICAM-1). The $IL-1{\beta}$ producing activity of THP-1 cells correlated well with the ability to induce the Mac-1 expression on THP-1 surface. Monoclonal antibody raised against relevant cell surface glycoproteins on THP-1 were tested for their ability to block the response of THP-1 to T cells. Antibody to Mac-1 only partially blocked $IL-1{\beta}$ production by THP-1, whereas antibodies to ICAM-1 and HLA-D did not. These data indicate that regulation of Mac-1 expression on THP-1 cells can alter the responsiveness of these cells to contact by activated T cells, however other unknown structures on the THP-1 cells may be involved in this process also.