• Journal of Pharmacopuncture
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• v.16 no.4
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• pp.36-42
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• 2013

Objectives: Shengmaisan (SMS) is a traditional Chinese medicine prescription widely used for the treatment of diverse organs in Korea. The interstitial cells of Cajal (ICCs) are pacemaker cells that play an important role in the generation of coordinated gastrointestinal (GI) motility. We have aimed to investigate the effects of SMS in the ICCs in the mouse small intestine. Methods: To dissociate the ICCs, we used enzymatic digestions from the small intestine in a mouse. After that, the ICCs were identified immunologically by using the anti-c-kit antibody. In the ICCs, the electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICCs. Results: The ICCs generated pacemaker potentials in the mouse small intestine. SMS produced membrane depolarization with concentration-dependent manners in the current clamp mode. Pretreatment with a $Ca^{2+}$ free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in the endoplasmic reticulum, stopped the generation of the pacemaker potentials. In the case of $Ca^{2+}$-free solutions, SMS induced membrane depolarizations. However, when thapsigargin in a bath solution was applied, the membrane depolarization was not produced by SMS. The membrane depolarizations produced by SMS were inhibited by U-73122, an active phospholipase C (PLC) inhibitors. Furthermore, chelerythrine and calphostin C, a protein kinase C (PKC) inhibitors had no effects on SMS-induced membrane depolarizations. Conclusions: These results suggest that SMS might affect GI motility by modulating the pacemaker activity through an internal $Ca^{2+}$- and PLC-dependent and PKC-independent pathway in the ICCs.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.127-137
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• 1994

This study was carried out to investigate the inhibiting or promoting effect of fetal bovine serum fractionated by the molecular weight and to examine the effect of reconstruction of serum fractions on the development of 1- and 2-cell mouse embryos fertilized in vitro (IVEE) . The serum was separated by ultrafiltration or gel filtration methods and added in m-KRB medium for culture of IVFE. The developemental ability(cavitation and hatching) of embryos following culture of day 4 and 6 was compared among fractions. Small molecular weight fraction( <3 kDa) significantly inhibited the development of 1-and 2-cell IVFE to the blastocyst stages, compared with other fractions. One-cell IVFE were more sensitively damaged than 2-cell embryos by that fraction and arrested mainly at 2~4 cell stages. Moreover, small amount(<3%,v /v) of the inhibiting fraction acted even with protein rich fraction(100~30 kDa) and arrested the embryonic development. On the other hand, 100~30 kDa fraction promoted the embryonic development and no inhibiting effect was observed at the level of 50%(v /v) in culture medium In the experiment of gel filtraton, =30 kDa fraction showed the highest promoting effect on the embryonic development, but <4 kDa fraction inhibited significantly the development. These results suggest that serum contains not only small molecular weight inhibitory component(s) but also promoting one rather than albumin on embryonic development. And serum can be more effectively used in the IVF program after removal of inhibitory component(s) by one of above separation methods.

• Journal of Pharmacopuncture
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• v.21 no.4
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• pp.268-276
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• 2018

Objectives: The purpose of this study is to investigate the anti-cancer effects of different fractions of Astragalus membranaceus (AM) in human non-small cell lung cancer (NSCLC) cells. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AM. The cell death was examined by MTT assay and trypan blue exclusion assay. Apoptosis was detected by DAPI staining, annexin V-PI double staining and cell cycle analysis. The expression of apoptosis-related proteins and mitogen-activated protein kinases (MAPKs) was examined by western blot. Results: Among various fractions of AM, the ethyl acetate fraction of AM (EAM) showed the strongest cytotoxic effect in NSCLC cells. EAM reduced the cell proliferation in a time- and dose-dependent manner in NSCLC cells. In addition, EAM induced the chromatin condensation, and increased the population of sub-G1 phase and annexin V-positive cells in a time-dependent manner, indicating that EAM induced apoptosis in NSCLC cells. Consistently, EAM enhanced the expression of cleaved caspase-8 and -9, and induced the accumulation of cleaved- poly (ADP-ribose) polymerase (PARP). Among MAPK proteins, only ERK was dephosphorylated by EAM, suggesting that ERK might be related with EAM-induced apoptosis. Conclusion: Our results clearly demonstrate that EAM exhibited anti-cancer effects in NSCLC cells by induction of apoptosis. We provide a valuable evidence which suggests that AM could be a desirable therapeutic option for treatment of NSCLC.

• Proceedings of the Korean Society of Crop Science Conference
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• 2023.04a
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• pp.157-157
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• 2023

Pre-harvest sprouting (PHS) in rice (Oryza sativa L.) is one of the main problems associated with seed dormancy. PHS causes yield loss and reduction of grain quality under unpredictable humid conditions at the ripening stage, thus affecting the economic value of the rice crop. To resolve this issue, it is important to understand the molecular mechanism underlying seed dormancy in rice. Recent studies have shown that seed dormancy is affected by a large number of genes associated with plant hormones. However, the effect of heat stress on seed dormancy and plant hormones is not well understood. In this study, we compared the PHS rate as well as the transcriptome and small RNAome of the seed embryo and endosperm of two different accessions of rice, PHS-susceptible rice (low dormancy) and PHS-resistant rice (high dormancy) under three different maturation stages. We identified and verified the candidate genes associated with seed dormancy and heat stress-related responses in rice using quantitative real-time PCR. We newly discovered hormone-related genes, heat shock protein-related genes, and miRNAs potentially involved in PHS. These findings provide a foundation for understanding the dynamics of transcriptome and small RNAome of hormone- and heat stress-related genes, which affect PHS during seed maturation.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.1020-1024
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• 2003

Five barrows (average initial body weight 58.6 kg) were used to determine the apparent ileal digestibilities of amino acids, DM, N and energy in various soybean meal, rapeseed meal and coconut meal in finishing pigs. Dietary treatments included 1) KSBM (Korean soybean meal), 2) CSBM (Chinese soybean meal), 3) ASBM (Argentine soybean meal), 4) RSM (Rapeseed meal), and 5) CNM (Coconut meal). The diets were corn starch-based and formulated so that each protein source provided the same amount of total ME (3,490 kcal/kg), CP (15.70%), lysine (1.00%), Ca (0.80%) and P (0.60%). Protein content of the KSBM was higher than the CSBM and ASBM, with all values similar to those expected, and protein content of the CNM was lower than that of the SBM preparation and RSM. Apparent ileal digestibilities of histidine, lysine, threonine, alanine, asparatic acid, cystine, glutamic acid and serine were greater for the KSBM, CSBM, ASBM and RSM than for the CNM (p<0.05). Also, the apparent ileal digestibilities of methionine, leucine, phenylalanine, valine and tyrosine were greater for the KSBM than for the CSBM, ASBM, RSM and CNM (p<0.05). Overall, the apparent ileal digestibilities of total essential amino acids were greater for the KSBM than for the CSBM, ASBM, RSM and CNM (p<0.05), and the apparent ileal digestibilities of total non essential amino acids were greater for the KSBM, CSBM, ASBM and RSM than for the CNM (p<0.05). No difference (p>0.05) in apparent digestibility of DM at the small intestine was observed among the treatments. However, the apparent digestibility of DM at the total tract was greater for the KSBM than for the CSBM, ASBM, RSM and CNM (p<0.05). Also, apparent digestibilities of N and digestible energy at the small intestine and total tract were greater for the KSBM than for the RSM and CNM (p<0.05). In conclusion, nutrient digestibility values of SBM preparations and RSM were relatively high compared to CNM.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.7
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• pp.2953-2958
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• 2015

Although associations between thioredoxin interacting protein (TXNIP) and cancers have been recognized, the effects of TXNIP on non-small cell lung cancer (NSCLC) prognosis remained to be determined in detail. In addition, while hypoxia is a key characteristic of tumor cell growth microenvironment, the effect of hypoxia on TXNIP expression is controversial. In this study, formaldehyde fixed and paraffin embedded (FFPE) samples of 70 NSCLC patients who underwent resection between January 2010 and December 2011 were obtained. Evaluation of TXNIP and hypoxia inducible factor-$1{\alpha}$ ($HIF-1{\alpha}$) protein expression in FFPE samples was made by immunohistochemistry. By Kaplan-Meier method, patients with high TXNIP expression demonstrated a significantly shorter progression free survival (PFS) compared with those with low TXNIP expression (18.0 months, 95%CI: 11.7, 24.3 versus 23.0 months, 95%CI: 17.6, 28.4, P=0.02). High TXNIP expression level was also identified as an independent prognostic factor by Cox regression analysis (adjusted hazard ratio: 2.46; 95%CI: 1.08, 5.56; P=0.03). Furthermore, TXNIP expression was found to be significantly correlated with $HIF-1{\alpha}$ expression (Spearman correlation=0.67, P=0.000). To further confirm correlations, we established a tumor cell hypoxic culture model. Expression of TXNIP was up-regulated in all three NSCLC cell lines (A549, SPC-A1, and H1299) under hypoxic conditions. This study suggests that hypoxia induces increased TXNIP expression in NSCLC and high TXNIP expression could be a poor prognostic marker.

• Advances in pediatric surgery
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• v.15 no.1
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• pp.1-10
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• 2009

Recently, amniotic fluid has gained attention as one of the potential sources for cell therapy and tissue engineering because it has characteristics of multipotent stem cells. However, current knowledge about what types of cells are naturally found in amniotic fluid is still limited. In this study, we aimed to investigate whether human amniotic fluid contains cells that have characteristics of respiratory cells. Samples of human amniotic fluid (5 mL per sample) obtained from amniocenteses were cultured with small airway growth medium (SAGM). Cells were grown until the third passage and the presence of type II alveolar cells were characterized by inverted microscopy, immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR). On inverted microscopy, cultured cells showed typical polygonal and cobblestone-like epithelial morphology. The morphology of cells was not changed after selection and passing. Immunofluorescence analysis demonstrated that the isolated cells stained positive for surfactant protein C (SPC), specific marker for type II alveolar cells. Cells also stained positive for TTF-1 protein but negative for CD 31 and vimentin. RT-PCR analysis of cells showed expression of SPC mRNA. This study has demonstrated that respiratory cells can be isolated and identified from human amniotic fluid cultured in SAGM medium. Our results may provide the basis for further investigations of amniotic fluid.

• Journal of the Korean Vacuum Society
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• v.14 no.3
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• pp.138-142
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• 2005

We tested performance of the 4C1 beamline for analyzing structures of proteins in solution using small angle X-ray scattering (SAXS) at the Pohang Light Source(PLS). Structurally well-known proteins such as lysozyme and $Bcl-XL(\vartriangle TM/\vartriangle loop)$ were used for the study. Low resolution solution structures of lysozyme and $Bcl-XL(\vartriangle TM/\vartriangle loop)$ were obtained at a resolution of at least i.2 nm, and the structures were basically same as those calculated from the crystal structures of the proteins. We also used $Bcl-XL(\vartriangle TM/\vartriangle loop)$ with a long flexible loop attached [$Bcl-XL(\vartriangleTM))$] and obtained significantly different data from $Bcl-XL(\vartriangle TM/\vartriangle loop)$, although the electron density map of the loop is known to be invisible from the crystal structure of $Bcl-XL(\vartriangleTM))$. We confirm that SAXS experiment is a powerful tool for the structural study of proteins in solution and the 4Cl beamline at the PLS is well-equipped and suitable for the protein solution SAXS experiment.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.485-489
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• 2002

To investigate the effects of the dissolved oxygen concentration (DO) of Tetillarca granosa (Linnaeus), We measured $LC_{50}$, survival, oxygen consumption rate, filtration rates and blood composition under $10{\cdot}{\pm}0.5^{\circ}C$ and $25{\cdot}0.5^{\circ}C$ as a function of DO. The 16 days-$LC_{50}$ of DO for T. granosa was 1.31 mg DO/L in large individual and 0.95 mg DO/L in small individual. At 25$^{\circ}C$, the $LC_{50}$ of DO in large and small individual was 1.13 and 1.M mg DO/L, respectively. With decreasing DO, oxygen consumption rate, and filtration rates of T. granosa decreased. Blood composition of T. granosa was analysed hemoglobin, glucose, total protein, total cholesterol, GOT and GPT, Hg was increased with decreasing DO, but glucose was decreased below 1.2 mg DO/L. Total protein, total cholesterol, GOT and GPT were investigated irregular and decreasing aspect.

• Journal of Life Science
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• v.19 no.11
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• pp.1506-1513
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• 2009

RGS proteins have been identified as negative regulators of G protein signalling pathways and attenuate the activity of GPCR receptors. However, information on the regulatory effects of RGS proteins in the activity of cannabinoid receptors is limited. In this study, the role of RGS proteins on the signal transduction of the CB2 cannabinoid receptor was investigated in HEK293 cells co-transfected with CB2-receptors and plasmids encoding RGS2, RGS3, RGS4 and RGS5. Treatment of cells with WIN55, 212-2, a CB2 receptor agonist, inhibited forskolin-induced cAMP response element (CRE) activity in CB2-transfected HEK293 (CB2-HEK293) cells. This inhibitory effect of WIN 55, 212-2 on CRE activity was reversed by co-transfection of CB2-HEK293 cells with RGS3, but not with RGS2, RGS4 and RGS5. However, endogenous RGS3 protein knocked down by a small interfering siRNA targeting RGS3 gene enhanced inhibition of forskolin induced CRE activity via agonist induced CB2 receptor signal transduction. These results indicate the functional role of endogenous RGS protein in cannabinoid signaling pathways and define receptor-selective roles of endogenous RGS3 in modulating CRE transcriptional responses to agonist induced CB2 receptor activity.