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Adenovirus-mediated expression of both antisense ODC and AdoMetDC inhibited colorectal cancer cell growth in vitro

Acta Pharmacologica Sinica 2006年第3期27卷 353-359页

作者： Bing ZHANG~2 Xian-xi LIU~(2,5) Yan ZHANG~2 Chun-ying JIANG~3 Qing-shan TENG~4 Hai-yan HU~2 Wei WANG~2 Lei GONG~2 ~2Institute of Biochemistry and Molecular Biology,School of Medicine.Shandong University,Jinan 250012,China ~3Department of Coloproctology,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China,~4Department of Neurosurgery.Cleveland Clinic Foundation,Cleveland,OH 44195,USA InstituteofBiochemistryandMolecularBiology SchoolofMedicineShandongUniversityJinan250012China DepartmentofColo-proctology AffiliatedHospitalofShandongUniversityofTraditionalChineseMedicineJinan250014China

Aim:To construct a recombinant adenovirus that can simultaneously expressboth antisense ornithine decarboxylase(ODC)and adenosylmethionine decar-boxylase(AdoMetDC)and detect its inhibitory effect on the intracellular polyaminepool and colorectal cancer cell ***:A 205-bp cDNA of AdoMetDCwas reverse-inserted into recombinant pAdTrack-ODCas vectors and recombinedwith pAdEasy-1 vectors in AdEasy-1 *** clones were selected andtransfected into the packaging cell HEK293 after they were linearized by *** fluorescent protein expression was used to monitor the process of adenovi-rus *** ODC and AdoMetDC protein levels were identified by west-ern blotting,and intracellular polyamine content was detected by reverse-phasehigh performance liquid chromatography.A viable cell count was used to deter-mine the growth of HT-29 cells with or without exogenous ***:Sequencing confirmed that AdoMetDC cDNA was successfully ligated into thepAdTrack-ODCas *** expression in 293 cells during virus packing andamplification was observed by fluorescence *** blotting dem-onstrated that both ODC and AdoMetDC were downregulated by Ad-ODC-AdoMetDCas,and consequently 3 kinds of polyamine(putrescine,spermidineand spermine)were reduced to very low ***-29 cell growth was signifi-cantly inhibited as compared with control conditions,and growth arrest was notreversed by exogenous ***:The successfully constructedrecombinant adenovirus,Ad-ODC-AdoMetDCas,blocked polyamine synthesisand has therapeutic potential for treating colorectal cancer in vitro.

关键词： polyamine ornithine decarboxylase adenosylmethionine decarboxylase adenovirus colorectal cancer

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Effect of L-Deprenyl on the Putrescine Level and Neuronal Damage after Transient Global Cerebral Ischemia in Gerbils

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International Journal of Organic Chemistry 2017年第2期7卷 171-184页

作者： Hyung Lee Yeun-Kyung Chu Joon-Ho Shon Kyung-Hee Chun Jee-In Kim Seong-Ryong Lee Department of Neurology School of Medicine and Brain Research Institute Keimyung University Daegu Korea School of Kinesiology Yeungnam University Gyeongsan Korea Department of Medical Education School of Medicine Yeungnam University Daegu Korea Department of Molecular Medicine Obesity-Mediated Disease Research Center School of Medicine Keimyung University Daegu Korea Department of Pharmacology Obesity-Mediated Disease Research Center School of Medicine Keimyung University Daegu Korea

L-Deprenyl is selective and irreversible monoamine oxidase B inhibitor, known to have neuroprotective properties. Putrescine, one of polyamine, is thought to be important in the neuronal cell damage associated with various type of excitatory neurotoxicity. We examined the effects of L-deprenyl on the changes in putrescine level and neuronal damage after transient global ischemia in ger-bils. Male Mongolian gerbils weighing 65 - 75 g were used in the experiment. Global ischemia was induced by occlusion of common carotid arteries for 3 min to observe neuronal injury in hippocampal pyramidal cells. L-Deprenyl group was given 10 mg/kg of L-deprenyl intraperitoneally immediately after, 3 h and 6 h after global ischemia. Treated animals were processed in parallel with ischemic animals receiving saline as a vehicle and with sham- operated controls. Hippocampal putrescine level was increased by global ischemia and inhibited by L-deprenyl treatment. In histological findings, counts of viable neurons were made in the pyramidal cell layer of the hippocampal CA1 area 3 days after ischemic insult. The number of viable neurons in the pyramidal cell layer of CA1 area was significantly increased in animals treated with L-deprenyl compared to vehicle-treated ischemic animals (p < 0.05). In terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick endlabeling (TUNEL) assay, semiquantitative analysis of dark-brown neuronal cells was made in the hippocampal CA1 area. There was also a significant difference in the degree of TUNEL staining in the hippocampal CA1 area between vehi-cle-treated and L-deprenyl-treated animals (p < 0.05). These data show L-deprenyl is effective as a prophylactic treatment for neuronal injury when it is administrated before ischemia but a further study need to know the effects of administration of L-deprenyl after ischemia and at given times after reper-fusion.

关键词： L-Deprenyl polyamine Global Ischemia Hippocampus Gerbil Neuroprotection

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IL-10 Gene Knockout Reduces the Expression of mGlu Receptor 1a/b and Decreases the Glutamate-Dependent Production of Nitric Oxide

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Journal of Biomedical Science and Engineering 2014年第13期7卷 1019-1029页

作者： Sopiko Koriauli Tamar Barbakadze Nino Natsvlishvili Nino Dabrundashvili Eka Kvaratskhelia David Mikeladze Ilia State University Tbilisi Georgia Ilia State University Tbilisi Georgia I. Beritashvili Center of Experimental Biomedicine Tbilisi Georgia Institute of Medical Biotechnology Tbilisi State Medical University Tbilisi Georgia

IL-10 provides trophic and survival effects directly on neurons, promotes axonal outgrowth, and stimulates neuroregeneration. In this study, we analyzed the activities of arginase and nitric oxide synthase (NOS) in synaptoneurosomes derived from brain cortex of C57BL/6 IL-10 gene-knockout (KO) and wild-type (Wt) mice and determined that the synaptoneurosomes derived from KO mice present lower arginase II activity and lower spermine content than those derived from Wt mice, whereas the basal NOS activity in the KO synaptoneurosomes was higher than that observed in the control synaptoneurosomes. Moreover, our results indicate that the plasma membranes isolated from the KO mice brain exhibit significantly lower spermine-induced enhancement of [3H] MK-801 binding than the plasma membranes from the brain of Wt mice. Glutamate increases the production of nitric oxide (NO) in Wt synaptoneurosomes in a dose-dependent manner, whereas in the KO synaptoneurosomes, this amino acid does not affect the synthesis of NO. The glutamate-dependent acceleration of NO synthesis in Wt synaptoneurosomes was abrogated by LY367385, an antagonist of mGluR1a/b. The western blot analysis of the synaptoneurosomal proteins demonstrates that the expression of the subunits of NMDAR (NMDAR2A and NMDAR2B), the level of NMDAR-bound nNOS and the expression of iNOS are not changed in KO mice and that only the level of mGluR1a/b is markedly reduced in the synaptoneurosomes of KO mice. We conclude that a neuroprotective and neuroregenerative property of IL-10, in addition to its effects on polyamine metabolism and the spermine-dependent modulation of NMDAR, may involve the regulation of mGluR1a/b expression.

关键词： IL-10 polyamine Biosynthesis Nitric Oxide Metabotropic Glutamate Receptor

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Cu(II) Extraction in Ionic Liquids and Chlorinated Solvents: Temperature Effect

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Green and Sustainable Chemistry 2011年第4期1卷 155-164页

作者： Raimondo Germani Maria V. Mancini Nicoletta Spreti Pietro Di Profio Gianfranco Savelli 不详

Room temperature ionic liquids have been currently used in liquid/liquid extraction processes in order to substitute conventional organic solvents. In this paper, a series of chlorinated solvents and 1-alkyl-3-me- thylimidazolium-based ionic liquids were selected to study the extraction efficiency of a lipophilic polya- mine 1,1,7,7-tetraethyl-4-tetradecyldiethylenetriamine (TE14DT) towards the model ion Cu(II) in such dif-ferent media. The effect of temperature on the extraction efficiency was also investigated. The metal ion par-tition was found to be strongly dependent on both the nature of the solvent and on the working temperature. The viscosity of ionic liquids and the water content in ionic liquid were found to affect the extraction effi-ciency of TE14DT. The chemical nature of the cation of ionic liquids, and in particular the alkyl chain length on imidazolium ring, also seemed to be important in determining the efficiency of the extraction process. Finally, preliminary experiments on back-extraction of Cu(II) ions from ionic liquid also revealed interesting hints to the development of a continuous transport process.

关键词： Chlorinated Solvents Liquid-Liquid Extraction Hydrophobic Ionic Liquids Lipophilic polyamine Temperature Effect

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Simple Detection Method of Biogenic Amines in Decomposed Fish by Intramolecular Excimer Fluorescence

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Food and Nutrition Sciences 2012年第7期3卷 1020-1026页

作者： Hirofumi Nishikawa Tatsuya Tabata Seiichi Kitani Health Service Center Tokyo University of Marine Science and Technology Konan Japan

Biogenic amines are known to have various biological functions such as not only neurotransmitter and cell proliferation but also food poisoning. Bacterially-decomposed amines such as histamine, agmatine, putrescine, cadaverine, spermidine and spermine cause allergic symptoms. We developed simple method for measurement of polyamine as indicator of food decomposition with a fluorometer by using 4-(1-Pyrene)butyric acid N-hydroxysuccinimide ester (PSE). PSE reacts with primary and secondary amino moieties of polyamines and produces the intramolecular excimer fluorescence. Excimer fluorescence with broad peak at around 470 nm was clearly detected in linear type biogenic amines such as putrescine, cadaverine, spermidine and spermine at 10 mM. However neither histamine nor trimethylamine altered the fluorescence. Decomposed sardine and mackerel by improper storage showed stronger intensity than fresh ones. Comparing with OPA-method, PSE method was useful for screening biogenic amines present in food, esp. fish since the analysis was simple after one-step purification procedure. An inexpensive system which can rapidly detect biogenic amines from food is necessary in a medium and small-sized food business. The technique using excimer has potential to realize the high through-put screening system for evaluation of food freshness and is expected to bring the public interests such as food security and safety of consumer.

关键词： 4-(1-Pyrene)Butyric acid N-Hydroxysuccinimide Ester PSE Excimer Fluorescence Biogenic Amine Fish Decomposition polyamine Food Poisoning

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High Concentration of Spermine Induces the Dedifferentiation of Somatic Cells into Pluripotent Stem Cells

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Journal of Biomedical Science and Engineering 2019年第2期12卷 92-110页

作者： Naofumi Shiomi School of Human Sciences Kobe College Nishinomiya Japan

Cancer tissues contain cancer stem cells (CSCs), which play important roles in cancer metastasis. However, the mechanisms through which cancer cells dedifferentiate into stem cells have not yet been elucidated. In this study, the effects of high concentrations of polyamines produced in cancer cells on dedifferentiation were examined. The results showed that when normal human fibroblasts were cultured with high concentrations of spermine, the obtained polyamine-induced cells expressed alkaline phosphatase and marker proteins of pluripotent stem cells, although apoptosis occurred in most cells. In contrast, another polyamine-induced stem (PIS) cell line (Spe-2 PIS cells), obtained by culture in medium containing Rock, p53, and Bax inhibitors plus spermine, did not show signs of apoptosis. These Spe-2 PIS cells expressed marker proteins of pluripotent stem cells and differentiated into cardiomyocytes, brown adipocytes, and nerve cells. These results suggest that a high concentration of spermine, which often induces apoptosis in normal cells, has the capacity to dedifferentiate somatic cells into pluripotent stem cells and may be associated with the dedifferentiation of cancer cells, which continuously produce high concentrations of spermine. Moreover, the procedure to obtain Spe-2 PIS cells, which is simple and efficient, may have potential applications in regenerative medicine.

关键词： polyamine Spermine Pluripotent Stem Cells Dedifferentiation Cancer Stem Cells Regenerative Medicine

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Spermidine mediated PLGA nanoparticles containing fluorofenidone for treatment of idiopathic pulmonary fibrosis

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长沙医学院学报 2019年第1期 28-47,23页

作者： Jing Tang Jianming Li Guo Li Haitao Zhang Ling Wang Dai Lid Jinsong Ding Key Laboratory breeding base of Hunan Oriented Fundamental and Applied Research of Innovative Pharmaceutics Changsha Medical University1501 Leifeng RoadChangsha 410219HunanPR China School of Pharmaceutical Sciences Central South University172 Tongzipo RoadChangsha 410013HunanPR China 17 Section 3South Renmin RoadChendu 610041SichuanPR China Xiangya Hospital Central South University87 Xiangya RoadChangsha 410008HumanPR China

Idiopathic pulmonary fibrosis(IPF) is a progressive, fatal lung disease with poor survival. The advances made in deciphering IPF have led to the approval of different anti-fibrotic molecules, such as pirfenidone and nintedanib. An increasing number of studies with particles(liposomes, nanoparticles, microspheres, nanopolymersomes and nanoliposomes) modified with different functional groups have demonstrated improvement in lung targeted drug delivery. In the present study, we prepared, characterized, and evaluated spermidine(SPD) modified poly-lactic-co-glycolide nanoparticles(SPD-AKF-PLGA/NPs) as carriers for fluorofenidone(AKF) in order to improve the anti-fibrotic efficacy of this drug in lung. SPD-AKF-PLGA/NPs were prepared and functionalized by modified solvent evaporation method with spermidine(SPD), poly-lactic-co-glycolide-polyethylene glycol(PEG-PLGA) groups. The size of SPD-AKF-PLGA/NPs was 172.5±4.3 nm. The AKF release from nanoparticles was shown to be fit Higuchi model. The A549 cellular uptake of the SPD-Cou-6-NPs group was found to be almost 2-fold higher than that of the Cou-6-NPs group. Free SPD and DFMO were pre-incubated in A549 cells to further prove that the uptake of SPD-Cou-6-PLGA/NPs via PTS. As a result, the uptake of SPD-Cou-6-PLGA/NPs significantly decreased with the increase of SPD concentrations in incubation. At the higher SPD concentrations in 50 and 500 μM, the uptake of SPD-Cou-6-PLGA/NPs reduced 0.34 and 0.49-fold than that without SPD pretreatment. After pretreatment with DFMO for 36 h, the cellular uptake of SPD-Cou-6-NPs reached 1.26-fold compared to untreated DFMO group. In biodistribution study, the drug targeting index of SPD-AKF-PLGA/NPs in lung was 3.62 and 4.66-fold than that of AKF-PLGA/NPs and AKF solution, respectively. It was suggested that the SPD-AKF-PLGA/NPs could accumulate effectively in the lung. The lung histopathology changes and collagen deposition were observed by HE staining and Masson staining in efficacy stu

关键词： Idiopathic Pulmonary fibrosis Fluorofenidone Spermidine polyamine transport system Nanoparticles

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The Effect of Curative Concentration on Thermal and Mechanical Properties of Flexible Epoxy Coated Jute Fabric Reinforced Polyamide 6 Composites

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Open Journal of Composite Materials 2012年第4期2卷 133-138页

作者： Smith Thitithanasarn Kazushi Yamada Umaru S. Ishiaku Hiroyuki Hamada Department of Advanced Fibro-Science Kyoto Institute of Technology Kyoto Japan Department of Textile Science and Technology Faculty of Science Ahmadu Bello University Zaria Nigeria

Many researchers have shown interest in the reinforcement of commodity thermoplastic with natural fibers. However, the drawback of natural fibers is their low thermal processing temperatures, that border around 200℃. In this investigation, we tried to improve the thermal stability of natural fibers with the use of flexible epoxy surface coating that could facilitate processing with engineering thermoplastics. Jute fabric and Polyamide 6 (PA6) composites were prepared by compression molding. The thermal decomposition characteristics of the jute fabric were evaluated by using thermo gravimetric analysis (TGA). Mechanical analysis was conducted to evaluate tensile test and three point bending test of composite. It was found that thermal degradation resistance of jute fabric was improved by coating with flexible epoxy resin. Moreover, the flexural modulus improved with increasing curative concentration. The interfacial interaction between the epoxy and PA6 was clearly indicated by the photo micrographs of the polished cross sections of the coated and uncoated jute fabric/PA6 composites.

关键词： Natural Fiber (Jute) polyamine 6 Flexible Epoxy Composites Thermal Degradation Mechanical Properties

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Identification and characterization of promoter in mouse spermine oxidase gene

Identification and characterization of promoter in mouse spe...

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中国生物化学与分子生物学会第十届会员代表大会暨全国学术会议

作者： Tang Guicheng Han Yu Wang Yanlin Institute of molecular biology Medical CollegeChina Three Gorges University

Spermine oxidase(SMO) is a FAD-dependent oxidase involved in polyamines(PA) homeostasis in mammalian *** enzyme can selectively catalyze degradation of *** the resulted products,H202 and 3-aminopropanal have potential to produce apoptosis and pathological *** this study,we proved that at least six transcriptional start points(+1,+4,+27,+31,+51,and +63) existed for SMO gene and all of them located in the same exon(exonl).A 2.1 kb DNA fragment(-2113+124) upstream the mouse SMO gene was obtained from NIH 3T3 cell and was demonstrated to have promoter activity. Multiple luciferase reporter plasmids were constructed from the original promoter-containing DNA fragment with pGL-2 basic as the *** gene analysis with these serial deletion promoters showed that the fragment between -176 and -25 was the core promoter region and the fragment between -615 and -176 was the important regulation region.

关键词： polyamine spermine oxidase promoter

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Cyclen-Based Lipids and Polymers as Gene Vectors

Cyclen-Based Lipids and Polymers as Gene Vectors

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第八届全国化学生物学学术会议

作者： Xiao-Qi Yu Key Laboratory of Green Chemistry and Technology（Ministry of Education） College of ChemistrySichuan University

＜正＞Gene therapy is a treatment method through the delivery of gene bounding to a vector and passing into *** with viral vectors,traditional nonviral vectors have some disadvantages such as low transfection efficiency and *** it is very important to develop some novel vectors which are safety and *** this presentation,a series of macrocyclic polyamine （MPA）-contained cationic lipids were designed and *** lipids were firstly applied

关键词： Cyclen gene delivery polyamine cationic lipids

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