• Clinical and Experimental Reproductive Medicine
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• v.37 no.2
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• pp.143-151
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• 2010

Objective: The aim of this study was to compare day 3 embryo transfer (D3ET) with day 5 ET (D5ET) in fresh in vitro fertilization (IVF) cycle on pregnancy outcomes. Methods: We conducted a retrospective matched case control study that included 90 women with D3ET and 90 women with D5ET from January 2007 to June 2009. Subjects were matched for reproductive profiles and IVF cycle characteristics. Two good quality embryos were transferred in both groups. Pregnancy rates (PR), implantation rate, and multiple PR were compared. Results: Demographics, stimulation parameters and embryological data were comparable in both groups. Main pregnancy outcomes with D3ET and D5ET groups were not statistically different: implantation rate (39.4% vs. 32.8%), positive PR (57.8% vs. 46.7%), clinical PR (53.3% vs. 45.6%), ongoing PR (50.0% vs. 42.2%), respectively. Both groups showed high multiple PR (37.5% vs. 34.1). Conclusion: D5ET may not be beneficial and necessary in comparison with D3ET on pregnancy outcomes, and elective single ET should be considered to decrease multiple pregnancies in women with favorable conditions and good quality embryos undergoing IVF.

• The Korean Journal of Quaternary Research
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• v.2 no.1
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• pp.25-50
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• 1988

The survival of rich evidence of palaeolithic occupation found in the Imjin-Hant'an River basin was possible due to many fortuitous geological conditions provided there. Formation of the basalt plain in a narrow valley system which developed during the late Mesozoic insured the appearance of a basin of sedimentation in which archaeological sites would be preserved with relatively minor post-depositional disturbance. Geomagnetic and K-Ar dating indicates that lava flows occurred during the Brunes Normal Epoch. During and after the process of basin sedimentation, erosion of the plain was confined to the major channel of the present river system which developed along the structural joints formed by the lava flow. Due to characteristic columnar structure and platy cleavage of the basalt bedrock, erosion of the basalt bedrock occurred mainly in vertical direction, developing deep but narrow entrenched valleys cut into the bedrock. Consequently, the large portion of the site area remained intact. Cultural deposits formed on top of the basalt plain were left unmodified by later fluvial disturbances due to changes in the Hant'an River base-level, since they were formed about 20 to 40m above the modern floodplain. Sedimentological evidence of cultural deposits and palynological analysis of lacustrine bed formed in the tributary basin of the Hant'an River indicate that hominid occupation occurred in this basin under rapidly deteriorating climatic conditions. From three thermoluminescence dates, the timing of hominid occupation as represented by 'Acheulian-like' bifaces apparently occur sometime during 45,000 BP. Thus, deposition of cultural layers in this basin approximately coincides with the beginning of the second stadial of the final glacial, during which the Korean Peninsula must have had provided a sanctuary for prolonged human occupation.

• Radiation Oncology Journal
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• v.18 no.3
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• pp.214-219
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• 2000

Purpose : To Investigate the pathways of radiation induced apoptosls and the effect of cysteamine (${\beta}$-mercaptoethyiamine), as a radioprotector, on it. Materials and Methods : HL-50 ceils were assigned to control, irradiated, and cysteamlne (1 mM, 10mM) pretreated groups. Irradiation was given In a single fraction of 10 Gy (6 MV x-ray) and cysteamine was administered 1 hour before irradiation. The activities of caspase-8 were measured in control and irradiated group to evaluate its relation to the radiation Induced apoptosis. To evaluate the role of cysteamine In radiation Induced apoptosis, the number of viable cells, the expression and activity of caspase-3, and the expression of poly (ADP-ribose) polymerase (PARP) were measured and compared after irradiating the HL-60 celis with cysteamine pretreatment or not. Results : The intraceliular caspase-8 activity, known to be related to the death receptor induced apoptosis, was not affected by irradiation(p>0.05). The number of viable cells began to decrease from 6 hours after irradiation (p>0.05), but the number of viable cells In 1 mM cysteamine pretreated group was not decreased after irradiation and was similar to those in the control group. In caspase-3 analyses, known as apoptosis executioner, its expression was not different but its activity was Increased by irradiation(p>0.05). However, this Increase of activity was suppressed by the pretreatment of 1 mM cysteamine. The cleavage of PARP, thought to be resulted from caspase-3 activation, occurred after irradiation which was attenuated by the pretreatment of 1 mM cysteamine. Conclusion : These results show that radiation induced apoptotic process is somewhat different from death receptor induced one and the pretreatment of 1 mM cysteamine has a tendency to decrease the radiation-induced apoptosis in HL-60 cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.29 no.5
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• pp.272-281
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• 2003

Nontraditional or alternative medicine is becoming an increasingly attractive approach for the treatment of various inflammatory disorders and cancers. Curcumin is the major constitute of turmoric powder extracted from the rhizomes of the plant Curcuma longa. Resveratrol is a phytoalexin present in grapes and a variety of medicinal plants. In this report, We investigated the effect of curcumin and resveratrol on regulatory protein of cell cycle, induction of apoptosis and MMP activity. Treatment with 75 M curcumin for 24 hrs produced morphological changing in HN-4 cells. Curcumin and resveratrol inhibited the cellular growth in HN-4 cells. Inhibition of cell growth was associated with down-regulation of cell cycle regulatory proteins. Curcumin-induced caspase-3 activation and Bax degradation were dose-dependent with a maximal effect at a concentration of 100 M. The elevated caspase-3 activity in curcumin treated HN-4 cells are correlated with down-regulation of survivin and cIAP1, but not cIAP2. Curcumin induced a dose-dependent increase of cytochrome c in the cytosol. Curcumin induced-apoptosis was mediated through the release of cytochrome c. In addition, curcumin-induced apoptosis was caused by the generation of reactive oxygen species, which was prevented by antioxidant N-acetyl-cysteine (NAC). Cotreatment with NAC markedly prevented cytochrome c release, Bax cleavage and cell death. Also resveratrol-induced apoptosis was preceded by down-regulation of the anti-apoptotic Bcl-2, cIAP1, and caspase-3 activity. However, resveratrol-induced apoptosis was not prevented by antioxidant NAC. In addition, HN-4 cells release basal levels of MMP2 when cultured in serum-free medium. Treatment of the cells with various concentrations of PMA for 24 hr induced the expression and secretion of latent MMP9 as determined by gelatin zymography. HN-4 cells were treated with various concentrations of curcumin and resveratrol in the presence of 75 nM PMA, and MMP2 and 9 activities were inhibited by curcumin and resveratrol. These findings have implications for developing curcumin-based anticancer and anti-inflammation therapies.

• Korean Journal of Clinical Pharmacy
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• v.13 no.1
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• pp.29-39
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• 2003

Neurodegenerative disorders are associated with apoptosis as a causing factor or an inducer. On the other hand, it has been reported that epigallocatechin gallate (EUG), one of antioxidants and flavonoids, and z-VAD-fmk, a nonselective caspase inhibitor, suppress oxidative-radical-stress-induced apoptosis. However, it is not yet known what is the effects of EGCG and z-VAD-fmk on the apoptotic pathway is through phosphoinositide 3-kinase (PI3K), Akt and glycogen synthase kinase-3 (GSK-3) as well as mitochondria, caspase-3 and poly (ADP-ribose) polymerase (PARP). We investigated the effects of EGCG by using $H_2O_2$ treated N18D3 cells, mouse DRG hybrid neurons. Methods: Following 30 min $100\;{\mu}m\;H_2O_2$ exposure, the viability of N18D3 cells (not pretreated vs. EGCG or z-VAD-fmk pretreated) was evaluated by using MTT assay. The effect of EGCG on immunoreactivity (IR) of cytochrome c, caspase-3, PARP, PI3K/Akt and GSK-3 was examined by using Western blot, and was compared with that of z-Y4D-fmk. Results: EGCG or z-VAD-fmk pretreated N18D3 cells showed increased viability. Dose-dependent inhibition of caspase-3 activation accompanied by PARP cleavage were demonstrated by pretreatment of both agents. However, inhibition of cytochrome c release was only detected in EGCG pretreated N18D3 cells. On the pathway through PI3K/Akt and GSK-3, however, the result of Western blot in EGCG pretreated N18D3 cells showed decreased IR of Akt and GSK-3 and increased IR of p85a PI3K, phosphorylated Akt and GSK-3, and contrasted with that in z-VAD-fmk pretreated N18D3 cells showing no changes on each molecule. Conclusion: These data show that EGCG affects apoptotic pathway through upstream signal including PI3K/Akt and GSK-3 pathway as well as downstream signal including cytochrome c and caspase-3 pathway. Therefore, these results suggest that EGCG mediated activation of PI3K/Akt and inhibition GSK-B could be new potential therapeutic strategy for neurodegenerative diseases associated with oxidative injury.

• The korean journal of orthodontics
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• v.28 no.2 s.67
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• pp.329-341
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• 1998

Sterilization has received much attention in orthodontic practices over the past several years. The present study was undertaken to investigate the effects of sterilization on the physical properties of orthodontic pliers-AEZ, Unitek, and Dentronix ligature cutters. This study was designed to examine the tips of ligature cutters before and after 200 and 400 sterilization cycles using the Bowmar RHT-1000, the Dentronix DDS-5000, and the Eschmann SES-2000. The tip surface and the fracture surface were observed with a scanning electron microscope. The microstructure was observed with an optical microscope. The hardness test was carried out with the mic개-Vickers hardness tester and the Rockwell C Scale hardness tester. The chemical composition was analyzed by energy dispersive X-ray spectrometer. The results of this study were as follows : 1. The number and the size of corrosion products on the tip surface and the proportion of cleavage planes in fractured specimen increased, but the hardness of the tip decreased in proportion to sterilization cycles. From these observations, it was considered that mechanical properities decreased in proportion to sterilization cycles. 2. The number and the size of chromium carbides increased in proportion to sterilization cycles. Coarse microstructure decreased mechanical properities. 3. The AEZ and Unitek ligature cutters were Fe-Cr stainless steels, but the Dentronix ligature cutter was Co-Cr alloy. There were many differences among manufactures, but the chemical composition was .not changed after sterilization cycles. 4. The tip edge of ligature cutter used in a clinic revealed microcracks with the SEM observation. Clinical experience confirmed that ligature cutters were gradually degraded by sterilization.

• Journal of Aquaculture
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• v.20 no.2
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• pp.96-105
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• 2007

A pair of maroon clownfishes with an indonesian native, reared in recirculation culture system to develope its aquaculture techniques. Courtship, spawning behavior, egg developments and rearing of the maroon clownfish larvae were documented. The larval development were described with illustrative figures. The spawning was occurred 8 times between Feburary and August 2004. The gravid female spawned during 15:00-20:00. The male mainly took care of the eggs supplying oxygen by water currents using their pectoral fins, anal fin and mouth. The fertilized eggs were separative-adhesive and oval in shape, and $1.99{\pm}0.03\;mm$ in longer diameter and $0.88{\pm}0.03\;mm$ in shorter diameter. The fertilized eggs were in deep-orange color. Cleavage occurred in 30 minutes after fertilization, and the egg reached 2 cells stage in 1 hour 10 minutes after fertilization at $27.0^{\circ}{\pm}0.5^{\circ}C$. The embryo was formed in 23 hours 40 minutes after fertilization. Hatching began in between $120{\pm}2$ hours and $150{\pm}12$ hours after fertilization at $27.0^{\circ}C$ in the incubator. Total length (TL) of the newly hatched larvae was 3.22 mm with mouth and anus opened. Ten days after hatching, mean TL of the larvae were 6.21 mm with 28 dorsal fin rays, 17 anal fin rays and 28 caudal fin rays. Nineteen days after hatching, mean TL of the larvae were 9.34 mm. At this stage the larva had three white bands on the body, and they began to feed on commercial diet.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• Journal of Life Science
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• v.18 no.11
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• pp.1499-1506
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• 2008

To examine antitumor activity of the edible plant Zanthoxylum schinifolium, the cytotoxic effect of various organic solvent extracts of its stems on human acute leukemia Jurkat T cells was investigated. Among these extracts such as methanol extract (SS-7), methylene chloride extract (SS-8), ethyl acetate extract (SS-9), n-butanol extract (SS-10), and residual fraction (SL-11), SS-8 exhibited the most cytotoxic activity against Jurkat T cells. The methylene chloride extract (SS-8) possessed the apoptogenic activity capable of inducing sub-G1 peak along with apoptotic DNA fragmentation in Jurkat T cells. Western blot analysis revealed that SS-8 induced apoptosis via mitochondrial cytochrome c release into cytoplasm, subsequent activation of caspase-9 and caspase-3, and cleavage of PARP, which could be blocked by overexpression of Bcl-xL. Jurkat T cell clone I2.1 $FADD^{-/-}$) and Jurkat T cell clone I9.2 (caspase-$8^{-/-}$ were as sensitive as was the wild-type Jurkat T cell clone A3 to the cytotoxic effect of SS-8, suggesting no contribution of Fas/FasL system to the SS-8-mediated apoptosis. The GC-MS analysis of SS-8 showed that it was composed of 16 ingredients including 9,12-octadecanoic acid (18.62%), 2,4-dihydro-5-methyl-4- (1-methylethylidene)- 2-(4-nitrophenyl)-3H- pyrazol-3-one (14.97%), hexadecanoic acid (14.23%), (z,z)-6,9-pentadecadien- 1-ol (13.73%), 5,6-dimethoxy-2-methyl benzofuran (10.95%), and 4-methoxy-2-methylcinnamic acid (5.38%). These results demonstrate that the methylene chloride extract of the stems of Z. schinifolium can induce apoptotic cell death in Jurkat T cells via intrinsic mitochondria-dependent caspase cascade regulated by Bcl-xL without involvement of the Fas/FasL system.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.55-61
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• 2009

Objective: To compare the clinical outcomes of ICSI with sperm retrieved from testicular tissue in patients with obstructive azoospermia (OA) or hypospermatogenesis (HS). Methods: From January 2003 through December 2006, 155 patients with OA (241 cycles) and 28 patients with HS (34 cycles) were included in this study. We compared clinical outcomes of ICSI with testicular sperm such as fertilization rate, implantation rate, clinical pregnancy rate and delivery rate. Data were statistically analyzed using t-test and ${\chi}^2$-test. Results: Testicular spermatozoa could not be retrieved in 1 out of the 21 cycles where fresh testicular sperm extraction in HS patients. Fertilization rate (FR) was significantly higher in OA than HS (75.6 % vs. 62.6%, p<0.001). Cleavage rate (CR) per fertilized zygote was also significantly higher in OA than that in HS (66.8% vs. 54.8% p<0.001). However, there were no significant differences in good embryo rate (GER), clinical pregnancy rate (CPR), implantation rate (IR) and delivery rate (DR). Conclusion: Our results show that testicular sperm of HS does not affect CPR, IR, and DR although it has shown reduced FR and CR.