• Title/Summary/Keyword: PC-12 Cell

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Relations of Platelet Indices with Endometrial Hyperplasia and Endometrial Cancer

  • Karateke, Atilla;Kaplanoglu, Mustafa;Baloglu, Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.4905-4908
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    • 2015
  • Background: Platelets are blood elements thought to play a role in the immune system and therefore tumor development and metastasis. Platelet activation parameters such as mean platelet volume (MPV), platelet distribution width (PDW), and plateletcrit (PCT) can be easily evaluated with the whole blood count and have been studied as markers of systemic inflammatory responses in various cancer types. Our aim in this study was to evaluate the correlation between endometrial pathologies and MPV, PDW and PCT. Materials and Methods: A total of 194 patients who presented to our clinic with abnormal vaginal bleeding were included in our study. The patients were divided into 3 groups (endometrial hyperplasia, endometrial cancer, control) according to their pathology results. The groups were compared for MPV, PDW, and PCT values obtained from the blood samples taken on endometrial biopsy day. Results: The endometrial cancer patients were the oldest group (p=0.04). There was no significant difference between the three groups in terms of white blood cell count (WBC), platelet count (PC), and hemoglobin (Hb) level. The highest MPV (p<0.001), PDW (p=0.002), and PCT (p<0.001) levels were in the endometrial cancer group, and the lowest levels were in the control group. Conclusions: The easy evaluation of platelet parameters in patients who are suspected of having endometrial pathology is a significant advantage. We found MPV, PDW, and PCT to be correlated with the severity of endometrial pathology with the highest values in endometrial cancer. Studies to be conducted together with different laboratory parameters will further help evaluate the diagnosis and severity of endometrial cancer and precursor lesions.

Tyrosinase Inhibitory Activity and Neuronal Cell Protection of Hydrothermal Extracts from Watermelons (수박 열수 추출물의 Tyrosinase 저해능과 신경세포 보호효과)

  • Heo, Da-Jeong;Kim, Su-Jung;Choi, Ae-Ran;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.10
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    • pp.1707-1711
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    • 2013
  • In our study, each part (flesh, white rind, and green rind) of watermelon was extracted using hydrothermal extraction method at temperatures ranging from 100 to $300^{\circ}C$ at the intervals of 10, 30, and 60 min. We found that hydrothermal treatment has a significant bearing not only on tyrosinase inhibitory activity but also on neuronal cell protection of watermelon parts. The peak tyrosinase inhibitory activity (about 93%) was observed in both the flesh and green rind extracts at $300^{\circ}C$ for 60 min. In addition, we observed that hydrothermal extracts of watermelon parts at $300^{\circ}C$ for 60 min also evidenced significant protection effect for neuronal cell against $H_2O_2$ in a concentrationdependent manner. The results of this study confirm that hydrothermal treatment may be an efficient processing method for the purpose of obtaining potent bioactive substances from watermelon.

CONSTRUCTION OF HNGF-$\beta$ RECOMBINANT ADENOVIRUS & SCREENING OF ITS EXPRESSION AFTER TRANSFECTION INTO VARIOUS CELL LINES (말초신경재생을 위한 hNGF-$\beta$ recombinant Adenovirus의 제작 및 수종세포주에서 신경성장인자의 발현)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Yoon-Tae;Park, Hee-Jung;Sung, Mi-Ae;Kim, Nam-Yeol;Yoo, Sang-Bae;Myoung, Hoon;Hwang, Soon-Jung;Kim, Myung-Jin;Kim, Sung-Min;Jang, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.5
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    • pp.446-456
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    • 2005
  • Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$\beta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$\beta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$\beta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$\beta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$\beta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$\beta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$\beta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$\beta$ gene transfected cells.

EFFECT OF NERVE GROWTH FACTOR GENE INJECTION ON THE NERVE REGENERATION IN RAT LINGUAL NERVE CRUSH-INJURY MODEL (백서 설신경 압박손상모델에서 신경성장인자 유전자 주입이 신경재생에 미치는 영향)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Soung-Min;Kim, Yun-Hee;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.375-395
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    • 2006
  • Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

Physiological Activity and Antiproliferation Effects of Citron Seed Extracts on Cancer Cells (유자씨 추출물의 생리활성과 암세포 성장 억제효과)

  • Lee, Yoon-Jeong;Hwang, In-Guk;Joung, Eun-Mi;Kim, Hyun-Young;Park, Eui-Seok;Woo, Koan-Sik;Jeong, Heon-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1672-1678
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    • 2009
  • This study was carried out to investigate the total polyphenol, total flavonoid content, antioxidant activity, angiotensin I-converting enzyme (ACE) inhibitory activity, $\alpha$-glucosidase inhibitory activity, and antiproliferation activity of the citron seed. The citron seed were separated to hull and embryo, and extracted with n-hexane and 70% ethanol. Antioxidant activity of ethanol extract was higher than that of n-hexane extract. IC50 value for DPPH radical scavenging activity of ethanol extract of hull (CSE1) and embryo (CSE2) were 3.18 and 8.43 mg/mL, and those of total antioxidant activity were 19.96 and 11.28 mg AA eq/g, respectively. ACE inhibitory activity and $\alpha$-glucosidase inhibitory activity on CSE1 showed the highest values of 31.61 and 45.17%, respectively. Antiproliferation effects on the MCF7, HepG2, H460, HCT-116, and PC3 cell line showed the highest values of 14.09, 19.12, 12.29, 9.78, and 9.12% in extract concentration of 5 mg/mL, respectively. These results suggested that citron seed can be used for development of functional food material which have biological activities.

Comparison of biological activities of essential oils from Foeniculum vulgare Mill, Boswellia carteii Birew and Juniperus rigida Sieb. by a supercritical fluid extraction system (초임계 추출 공법을 이용해 회향, 유향 및 노간주나무로부터 분리한 정유 성분의 생리활성 비교)

  • Lee, Hyun-Soo;Mun, Chul-Hyung;Park, Jin-Hong;Kim, Dae-Ho;Yoo, Jae-Eun;Park, Young-Sik;Ryu, Lee-Ha;Choi, Keun-Pyo;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.2
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    • pp.115-121
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    • 2003
  • Essential oils from Fennel fruit(Foeniculum vulgare Mill), Olibanum resin(Boswellia carteii Birew) and Needl Juniperus stem(Juniperus rigida Sieb.) were extracted by a supercritical fluid extraction system(SFE) and biological activity of each essential oils were observed. SFE technique was applied for the isolation and purification of nonpolar biologically active essential oils from each samples. The quantitative analysis of essential oils was carried out by gas chromatography-mass spectrometer(GC/MS). About 60% of the growth of AGS and A549 cells were inhibited by adding 1.0g/l of the crude essential oils and below 40% was observed by the control. Cytotoxicity on human normal lung cell(HEL299) was scored as $15{\sim}18%$ for the crude essential oils and 12% for control, respectively. It meant that the essential oils were more effective than the control in anti-mutagenecity tested by CHO V79 cells. The effect of the essential oils on the growth of nerve cells, PC12 was observed as follows: The viable cell density was about two times higher than control.

Cloning of cDNA Encoding Putative Cellular Receptor Interacting with E2 protein of Hepatitis C Virus (C형 간염바이러스 E2 단백질에 결합하는 추정 세포수용체 cDNA의 클로닝)

  • 이성락;백재은;석대현;박세광;최인학
    • Journal of Life Science
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    • v.13 no.4
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    • pp.541-550
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    • 2003
  • E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.

In vitro screening of the acetylcholinesterase inhibition, antioxidant activity, and neuronal cell protective effect of medicinal plant extracts (생약추출물의 acetylcholinesterase 저해, 항산화 및 신경세포보호 효과 in vitro 탐색)

  • Um, Min Young;Ha, Tae Youl;Seong, Ki Seung;Kim, Yong Sik
    • Food Science and Preservation
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    • v.20 no.6
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    • pp.840-845
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    • 2013
  • This study investigated the effects (i.e., the acetylcholinesterase activity, lipid peroxidation, and neuronal survival) of 20 kinds of medicinal water extracts. The water extracts of three medicinal plants - Cornus officinalis, Glycyrrhiza glabra, and Angelica gigas - were found to be the most effective on acetylcholinesterase inhibitory activity. In the lipid peroxidation-generating system induced by $H_2O_2/FeSO_4$ in rat brain homogenates, Perilla frutescens, Polygonum multiflorum, Cinnamomun cassia, and G. glabra exhibited protective activity against lipid peroxidation. The neuronal cell death induced by L-glutamate in PC12 was suppressed by the water extracts of G. glabra, Cinnamomun cassia, Platycodon grandiflorum, and Mentha arvensis at the concentration of $100{\mu}g/mL$. Taken together, these results showed that the water extract of G. glabra has the potential anti-dementia activity, which suggests that it might provide an effective strategy for improving dementia.

Transfer of Porcine Embryos Injected with Sperm Carrying with Exogenous DNA

  • Cho, Seong-Keun;Cho, Hwang-Yun;Park, Mi-Ryung;Park, Jong-Sik;Yoo, Jae-Gyu;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.61-61
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    • 2001
  • The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.

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Synthesis and Characterization of Quinoxaline-Based Thiophene Copolymers as Photoactive Layers in Organic Photovoltaic Cells

  • Choi, Yoon-Suk;Lee, Woo-Hyung;Kim, Jae-Ryoung;Lee, Sang-Kyu;Shin, Won-Suk;Moon, Sang-Jin;Park, Jong-Wook;Kang, In-Nam
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.417-423
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    • 2011
  • A series of new quinoxaline-based thiophene copolymers (PQx2T, PQx4T, and PQx6T) was synthesized via Yamamoto and Stille coupling reactions. The $M_ws$ of PQx2T, PQx4T, and PQx6T were found to be 20,000, 12,000, and 29,000, with polydispersity indices of 2.0, 1.2, and 1.1, respectively. The UV-visible absorption spectra of the polymers showed two distinct absorption peaks in the ranges 350 - 460 nm and 560 - 600 nm, which arose from the ${\pi}-{\pi}^*$ transition of oligothiophene units and intramolecular charge transfer (ICT) between a quinoxaline acceptor and thiophene donor. The HOMO levels of the polymer ranged from -5.37 to -5.17 eV and the LUMO levels ranged from -3.67 to -3.45 eV. The electrochemical bandgaps of PQx2T, PQx4T, and PQx6T were 1.70, 1.71, and 1.72 eV, respectively, thus yielding low bandgap behavior. PQx2T, PQx4T, and PQx6T had open circuit voltages of 0.58, 0.42, and 0.47 V, and short circuit current densities of 2.9, 5.29 and 9.05 mA/$cm^2$, respectively, when $PC_{71}BM$ was used as an acceptor. For the solar cells with PQx2T-PQx6T:$PC_{71}BM$ (1:3) blends, an increase in performance was observed in going from PQx2T to PQx6T. The power conversion efficiencies of PQx2T, PQx4T, and PQx6T devices were found to be 0.69%, 0.73%, and 1.80% under AM 1.5 G (100 mW/$cm^2$) illumination.