• Title/Summary/Keyword: TdT

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Valeriana jatamansi Jones Inhibits Rotavirus-Induced Diarrhea via Phosphatidylinositol 3-Kinase/Protein Kinase B Signaling Pathway

  • Zhang, Bin;Wang, Yan;Jiang, Chunmao;Wu, Caihong;Guo, Guangfu;Chen, Xiaolan;Qiu, Shulei
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1115-1122
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    • 2021
  • Rotavirus (RV), as the main cause of diarrhea in children under 5 years, contributes to various childhood diseases. Valeriana jatamansi Jones is a traditional Chinese herb and possesses antiviral effects. In this study we investigated the potential mechanisms of V. jatamansi Jones in RV-induced diarrhea. MTT assay was performed to evaluate cell proliferation and the diarrhea mice model was constructed using SA11 infection. Mice were administered V. jatamansi Jones and ribavirin. Diarrhea score was used to evaluate the treatment effect. The enzyme-linked immunosorbent assay was performed to detect the level of cytokines. Western blot and quantitative reverse transcription-PCR were used to determine protein and mRNA levels, respectively. Hematoxylin-eosin staining was applied to detect the pathological change of the small intestine. TdT-mediated dUTP nick-end labeling was conducted to determine the apoptosis rate. The results showed V. jatamansi Jones promoted MA104 proliferation. V. jatamansi Jones downregulated phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in protein level, which was consistent with the immunohistochemistry results. Moreover, V. jatamansi Jones combined with ribavirin regulated interleukin-1β (IL-1β), interferon γ, IL-6, tumor necrosis factor α, and IL-10, and suppressed secretory immunoglobulin A secretion to remove viruses and inhibit dehydration. V. jatamansi Jones + ribavirin facilitated the apoptosis of small intestine cells. In conclusion, V. jatamansi Jones may inhibit RV-induced diarrhea through PI3K/AKT signaling pathway, and could therefore be a potential therapy for diarrhea.

Short-Term Changes in Gut Microflora and Intestinal Epithelium in X-Ray Exposed Mice

  • Tsujiguchi, Takakiyo;Yamaguchi, Masaru;Yamanouchi, Kanako
    • Journal of Radiation Protection and Research
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    • v.45 no.4
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    • pp.163-170
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    • 2020
  • Background: Gut microflora contributes to the nutritional metabolism of the host and to strengthen its immune system. However, if the intestinal barrier function of the living body is destroyed by radiation exposure, the intestinal bacteria harm the health of the host and cause sepsis. Therefore, this study aims to trace short-term radiation-induced changes in the mouse gut microflora-dominant bacterial genus, and analyze the degree of intestinal epithelial damage. Materials and Methods: Mice were irradiated with 0, 2, 4, 8 Gy X-rays, and the gut microflora and intestinal epithelial changes were analyzed 72 hours later. Five representative genera of Actinobacteria, Firmicutes, and Bacteroidetes were analyzed in fecal samples, and the intestine was pathologically analyzed by Hematoxylin-Eosin and Alcian blue staining. In addition, DNA fragmentation was evaluated by the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Results and Discussion: The small intestine showed shortened villi and reduced number of goblet cells upon 8 Gy irradiation. The large intestine epithelium showed no significant morphological changes, but the number of goblet cells were reduced in a radiation dose-dependent manner. Moreover, the small intestinal epithelium of 8 Gy-irradiated mice showed significant DNA damaged, whereas the large intestine epithelium was damaged in a dose-dependent manner. Overall, the large intestine epithelium showed less recovery potential upon radiation exposure than the small intestinal epithelium. Analysis of the intestinal flora revealed fluctuations in lactic acid bacteria excretion after irradiation regardless of the morphological changes of intestinal epithelium. Altogether, it became clear that radiation exposure could cause an immediate change of their excretion. Conclusion: This study revealed changes in the intestinal epithelium and intestinal microbiota that may pave the way for the identification of novel biomarkers of radiation-induced gastrointestinal disorders and develop new therapeutic strategies to treat patients with acute radiation syndrome.

Apoptosis and autophagy of muscle cell during pork postmortem aging

  • Chunmei Li;Xialian Yin;Panpan Xue;Feng Wang;Ruilong Song;Qi Song;Jiamin Su;Haifeng Zhang
    • Animal Bioscience
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    • v.37 no.2
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    • pp.284-294
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    • 2024
  • Objective: Pork is an important source of animal protein in many countries. Subtle physiochemical changes occur during pork postmortem aging. The changes of apoptosis and autophagy in pork at 6 h to 72 h after slaughter were studied to provide evidence for pork quality. Methods: In this article, morphological changes of postmortem pork was observed through Hematoxylin-eosin staining, apoptotic nuclei were observed by TdT-mediated dUTP nick end labeling assay, protein related to apoptosis and autophagy expressions were tested by western blot and LC3 level were expressed according to immunofluorescence assay. Results: In this study, we found the occurrence of apoptosis in postmortem pork, and the process was characterized by nucleus condensation and fragmentation, formation of apoptotic bodies, increase in apoptosis-related Bax/Bcl-2 levels, and activation of caspases. Autophagy reached its peak between 24 and 48 h after slaughter, accompanied by the formation of autophagosomes on the cell membrane and expression of autophagy-related proteins beclin-1, P62, LC3-I, LC3-II, and ATG5. Conclusion: Obvious apoptosis was observed at 12 h and autophagy reached its peak at 48 h. The present work provides the evidence for the occurrence of apoptosis and autophagy during postmortem aging of pork. In conclusion, the apoptosis and autophagy of muscle cells discovered in this study have important implications for pork in the meat industry.

Effects of Physical Parameters on Water Quality in Agricultural Reservoirs (농업용 저수지의 물리적 인자가 수질에 미치는 영향)

  • Jeon, Ji-Hong;Ham, Jong-Hwa;Kim, Ho-Il;Hwang, Soon-Jin;Yoon, Chun-Gyeong
    • Korean Journal of Ecology and Environment
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    • v.35 no.1 s.97
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    • pp.28-35
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    • 2002
  • The effect of physical parameters on water quality was analyzed using monitoring data of 193 agricultural reservoirs. The retention time of reservoirs ($t_d$) ranged between 10 and 140 days, and the ratio of drainage area (DA) to reservoir surface area (SA) was between 10 and 120. Both ratios of DA/SA and total area (TA)/ reservoir storage (ST) in Korean agricultural reservoirs were relatively greater than those in natural lakes in other countries. As retention time was plotted against DA/SA ratio, it was shorter in Korean reservoirs than natural lakes. The semi-logarithmic relationship between TA/SA and t>$t_d$ was $t_d\;=\;42.21(TA/ST)^{-1}$ (n = 50, $R^2\;=\;0.89$). While areal loading of total phosphorus (TP) was below $4\;gTP{\cdot}m^{-2}{\cdot}yr^{-1}$ in general, it exceeded $10\;gTP{\cdot}m^{-2}{\cdot}yr^{-1}$ in reservoirs where DA/SA ratio was greater than 100, which implies that areal loading of TP increases as DA/SA ratio increases. Chl-a concentration was positively related with the mean depth of reservoir, implying the higher Chl-a concentration with deeper the mean depth. Therefore, the deeper reservoir might be advantageous in water quality management perspective if other morphological conditions are similar. The empirical regression equation using physical parameters was also suggested in the estimation of TP concentration in the reservoirs. Combined information presented in this paper might be applicable to the water quality management in agricultural reservoirs.

Selection of Representative Foods and 'Best-fit' Mapping of Other Foods for Estimation of a Comprehensive Exposure to Food Contaminants in a Korean Total Diet Study (한국인의 유해물질 식이 노출량 평가를 위한 대표식품 선정과 mapping)

  • Koh, Eun-Mi;Shin, Hye-Hyung;Yon, Mi-Yong;Nam, Eun-Jeong;Lee, Yoon-Na;Kim, Do-Hee;Lee, Jee-Yeon;Kim, Mee-Hye;Park, Sung-Kug;Choi, Hoon;Kim, Cho-Il
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.773-782
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    • 2011
  • A food list representing the typical diet of Koreans was developed for a total diet study based on the 687 food items which appeared in the dietary intake data collected during the 2008 & 2009 Korea National Health & Nutrition Examination Survey. Food items included in 90% of the cumulative intake, items with a consumption frequency of 5% or higher and items contributing to 80% of cumulative fat intake were considered first. From the resulting 165 food items, 15 items were removed to avoid similar foods, while 10 distinct items were added to increase the efficiency of 'best-fit' mapping. Consequently, 160 items were selected as representative foods, which cover 90.93% of intake amount, 91.36% of energy intake and 89.05% of fat intake. Given that every food consumed by Koreans can't be analyzed for contaminants of interest, mapping other foods to the previously mentioned representative 160 foods is warranted because it enables a more comprehensive dietary exposure assessment that is close to reality.

Attenuation of Reperfusion Injury with Angiotension $AT_1$ Receptor Blockade in Rat Myocardial Ischemic Model (백서 심근 허혈 모델에서 angiotension $AT_1$수용체 차단제의 재관류 손상 감소 효과)

  • Choi, Jun-Young;Choi, Dong-Ju;Ahn, Hyuk
    • Journal of Chest Surgery
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    • v.34 no.3
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    • pp.203-211
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    • 2001
  • 배경: AT$_1$수용체의 길항제가 세포 수준에서 심근을 재관류 손사으로부터 보호할수 있다는 것으로 알려져 있지만, 생체내에서의 효과나 그 기전은 아직 명확히 밝혀지지 않았다. 본 연구에서는 백서 심근 허혈 모델을 이용하여, AT$_1$ 수용체의 길항제들 중 하나인 irbesartan이 심근이 재관휴 손상에 미치는 효과를 알아보고, 재관류 손상을 매개하는 한 각지 기전으로서 세포자멸의 기여에 대하여 연구하고자 하였다. 대상 및 방법: Sprague-Dawley 백서에서 무작용 부형약(10% gum arabic: 1군, 개체수=14관) irbesartan(50mg/kg/day :II 군, 개체수=12)을 각각 3일 동안 24시간마다 경구로 투여하였다. 실험동물의 좌 관상 동맥을 45분간 결찰하였다가, 그 후 2시간 동안 재관류시킨 다음 심장을 적출 하였다. TTC(triphenyltetrazolium chloride) 염색법을 이용하여, 허혈 노출 부위에 대한 심근 경색 부위의 비율을 측정하였다. Agarose gel 전기영동상의 DNa 분절 양상과 TUNEL(TdT-mediated dUCP nick end labeling) 염색을 관찰하여 세포자멸이 일어난 정도를 평가하였다. 세포자멸을 조절하는데 관여하는 것으로 알려진 Bcl-2(B-cell lymphoma 2 gene), Bad 등의 단백과 ERK (extracellular signal-regulated kinase), p-38 등 신호전달체계에 작용하는 MAPKs(mitogen-activated protein kinases)의 발현을 측정하기 위하여 Western blot을 시행하였다. 결과: 허혈 노출부위에 대한 심근 경색부위의 비율은 II군(42$\pm$2.7%)이 I군( 64.1$\pm$4.65)에 비해 유의하게 작았다.(p< 0.05), Agarose gel 전기영동상의 DNA laddering 양상은 I군에서 보다 높게 발현되었다. Bad와 ERK2의 발현은 두 군간에 유의한 차이가 없었다. 결론: AT$_1$수용체 길항제인 irbesartan은 생체에서 심근의 재관류 손상을 줄이는 효과가 있었다. 이 효과는 적어도 부분적으로 나만 심근세포의 세포자멸이 감소한 것에 기인한 것으로 설명할 수 있으며, 이 항-세포 자멸 효과는 Bcl-2의 발현증가와 관련이 있는 것으로 추정되었다.

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Apoptosis and Cell Proliferation in Gastric Adenoma and Adenocarcinoma (위샘종과 위샘암종에서의 세포자멸사와 세포증식)

  • Lee, Dong-Soo;Kang, Sang-Bum;Lee, Seung-Woo;Nam, Soon-Woo;Yoo, Young-Kyung;Han, Sok-Won
    • Journal of Gastric Cancer
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    • v.6 no.2
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    • pp.91-96
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    • 2006
  • Purpose: Cancer is a genetic disease caused by alterations in key regulators of cell growth and cell turnover, We investigated apoptotic cell death and cell proliferation in gastric adenomas and adenocarcinomas. Materials and Methods: The TdT-mediated dUTP-biotin nick end labelling (TUNEL) method and immunohistochemistry for Ki-67 were peformed, using paraffin-embedded tissues of 41 gastric adenomas and 100 gastric adenocarcinomas. These results were compared with histopathologic parameters. Results: The Ki-67 labelling index was higher in adenocarcinomas than in adenomas and the apoptotic index was higher in adenomas than in adenocarcinomas. There were no significant difference between the apoptotic index/Ki-67 labelling index and clinicopathological parameters. Conclusion: We propose that cell proliferation is more closely associated with gastric adenocarcinomas than apoptosis is, but that neither has any clinical significance as a prognostic factor in gastric adenocarcinomas. (J Korean Gastric Cancer Assoc 2006;6:91-96)

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Experimental Study of Endostar Injection Concomitant with Cryoablation on Lung Adenocarcinoma A549 Xenografts

  • Ma, Chun-Hua;Jiang, Rong;Li, Jin-Duo;Wang, Bin;Sun, Li-Wei;Lv, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6697-6701
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    • 2013
  • Objective: To explore the inhibiting effect and mechanism of Endostar injection concomitant with cryoablation on lung adenocarcinoma A549 xenografts in nude mice. Materials and Methods: A total of 24 nude mice with subcutaneous xenografts of the A549 cell line were established and divided into 4 groups when the maximal diameters of tumors became 1 cm: control group, Endostar group, cryoablation group and combination group (Endostar concomitant with cryoablation). The nude mice were sacrificed after 21-days treatment, tumour tissues were removed to measure their volume, in situ test of TdT-mediated dUTP nick end labeling (TUNEL) was adopted to determine the cellular apoptosis around freezing injury zones, and immunohistochemical SP test was applied for the detection of micro-vessel density (MVD) and vascular endothelial growth factor (VEGF) expression levels. Results: At 21-days after treatment, the growth velocities of control group, Endostar group, cryoablation group and combination group were $236.7{\pm}51.2%$, $220.0{\pm}30.6%$, $159.5{\pm}29.3%$ and $103.3{\pm}25.5%$ (P<0.01), while cellular apoptosis rates of tumors were $21.7{\pm}2.34%$, ($22.17{\pm}1.47$)%, $38.3{\pm}1.37%$ and $49.2{\pm}1.72%$, (P<0.01), respectively, according to the immunohistochemical test. MVD and VEGF expression levels in the combination group were both lower than in other groups (P<0.01), also being positively related (r=0.925, P<0.01). Conclusions: Endostar can significantly improve the inhibitory effects of cryoablation on xenografts of lung adenocarcinoma A549, and the mechanism is probably associated with its function as an inhibitor of tumour neo-angiogenesis through down-regulating VEGF expression.

Effects of Reactive Oxygen Species on Sperm Function, Lipid Peroxidation and DNA Fragmentation in Bovine Spermatozoa (소 정자에 있어서 활성산소계가 정자 기능과 지방산화 및 DNA 절편화에 미치는 영향)

  • Ryu, Buom-Yong;Chung, Yung-Chai;Kim, Chang-Keun;Shin, Hyun-A;Han, Jung-Ho;Pang, Myung-Geol;Oh, Sun-Kyung;Kim, Seok-Hyun;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.105-115
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    • 2002
  • Objective : To evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase (XO) system on sperm function, the change of sperm characteristics, lipid peroxidation, and DNA fragmentation in bovine spermatozoa. Materials and Methods: ROS were produced using a combination of 1000 uM X and 50 mU/ml XO. The ROS scavengers: superoxide dismu tase (SOD) (200 U/ml) and catalase (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without ROS scavengers at $37^{circ}C$ under 5% $CO_2$ incubator. Sperm movement characteristics by CASA (computer-aided sperm analysis), HOST (hypoosmotic swelling test), Caionophore induced acrosome reaction, malondialdehyde formation for the analysis of lipid peroxidation, the percentage of DNA fragmentation using the method of TdT-mediated nick end labelling (TUNEL) by flow cytometry were determined after 2 hours incubation. Results: The action of ROS on bovine spermatozoa resulted in a decreased in capacity for sperm motility, Ca-ionophore induced acrosome reaction and membrane integrity, an increased in malondialdehyde formation and the percentage of sperm with DNA fragmentation. In the effects of antioxidant, catalase completely alleviated the toxic effects induced by the ROS in terms of sperm function and characteristics, however SOD exhibited no capacity to reduce the toxic effects. Conclusion: The ROS can induce significant damages to sperm functions and characteristics. The useful ROS scavengers can minimized the defects of sperm function and various damages of spermatozoa.

Synthesis and Photovoltaic Properties of Conjugated Polymers Having Push-pull Structure according to the Type of Side-chain in the N-Substituted Phenothiazine (Push-pull 구조의 공액 고분자 합성 및 Phenothiazine의 질소 원자에 치환된 Side-chain에 따른 유기박막태양전지로의 특성 연구)

  • Seong, Ki-Ho;Yun, Dae-Hee;Woo, Je-Wan
    • Applied Chemistry for Engineering
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    • v.25 no.6
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    • pp.624-631
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    • 2014
  • In this study, a new series of conjugated polymer 3-(5-(5,6-bis(octyloxy)-7-(thiophen-2-yl)benzo[c][1,2,5]thiadiazol-4-yl)thiophen-2-yl)-10-(4-(octyloxy)phenyl)-10H-phenothiazine (P1) and 3-(5-(5,6-bis(octyloxy)-7-(thiophen-2-yl)benzo[c][1,2,5]thiadiazol-4-yl)thiophen-2-yl)-10-(4-((2-ethylhexyl)oxy)phenyl)-10H-phenothiazine (P2) were synthesised and organic photovoltaics (OPVs) properties were characterized. The push-pull structure polymer consisted of phenothiazine derivative as an electron donor and benzothiadiazole derivative as an electron acceptor. The aliphatic chain substituted aromatic ring was substituted at the position of N in phenothiazine for the electron-rich and improved solubility. Excellent thermal stabilities of P1 and P2 were confirmed by measured Td values as 321.9 and $323.7^{\circ}C$, respectively and the degrees of polymerization were 4,911 (P1) and 5,294 (P2). The maximum absorption wavelength of P1 and P2 were 549 and 566 nm, respectively. The device was fabricated and the OPVs property was measured. As a result, the power efficiency of conversion for P1 and P2 were 0.96 and 0.90%, respectively.