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Brain-derived neurotrophic factor genes transfect rat bone marrow mesenchymal stem cells based on cationic polymer vector

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Neural Regeneration Research 2009年第1期4卷 26-30页

作者： Zunsheng Zhang Kun Zan yonghai liu Xia Shen Department of Neurology Affiliated Hospital of Xuzhou Medical Collage Xuzhou 221009 Jiangsu Province China

BACKGROUND： Gene therapy is an effective expression of genes within target cells after transferring exogenous target genes. Both vector selection and transfection method are important factors for gene transfection. An ideal gene vector is required for a high transfusion of target gene and an exact introduction of target gene into specific target cells so as to express gene products. OBJECTIVE： To study the expression of mRNA and protein after transfecting rat bone marrow mesenchymal stem cells （BMSCs） with brain-derived neurotrophic factor （BDNF） genes based on cationic polymer vector. DESIGN, TIME AND SETTING： A randomized, controlled in vitro study using gene engineering, performed at the Neurobiology Laboratory, Xuzhou Medical College between October 2007 and April 2008. MATERIALS： PcDNA3.1 BDNF was obtained from Youbiai Biotechnological Company, Beijing and cationic polymer vector used was the SofastTM gene transfection reagent that was made by Taiyangma Biotechnological Co., Ltd., Xiamen. METHODS： BMSCs extracted from six Sprague Dawley （SD） rats aged 1 month were isolated and cultured in vitro. Third passage BMSCs were inoculated on a 6-well culture plate at the density of 1×106 cells/L. At about 80% confluence, BMSCs were transfected with PcDNA3.1-BDNF （2 μg） combined with SofastTM gene transfection reagent （6 μg）（BDNF group） or with PcDNA3.1 （2 μg） combined with SofastTM gene transfection reagent （6 μg）（blank vector group）. Cells that were not transfected with any reagents but still cultured under primary culture conditions were used as a non-transfection group. MAIN OUTCOME MEASURES： Enzyme linked immunosorbent assay was used to measure time efficiency of BMSC-secreted BDNF protein. Twenty-four hours after gene transfection, RT-PCR was used to detect expression of BDNF mRNA in the BMSCs. Immunohistochemistry was used to determine expression of BDNF protein in the BMSCs. RESULTS： BDNF protein expression was detected at day 1 after gene transfec

关键词： bone marrow mesenchymal stem cells brain-derived neurotrophic factor gene transfection

Synergistic effects of brain-derived neurotrophic factor and retinoic acid on inducing the differentiation of bone marrow stromal cells into neuron-like cells in adult rats in vitro

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Neural Regeneration Research 2006年第4期1卷 301-303页

作者： yonghai liu Yucheng Song Zunsheng Zhang Xia Shen Department of Neurology Affiliated Hospital of Xuzhou MedicalCollege Xuzhou 221002Jiangsu Province China

BACKGROUND： Under induction of retinoic acid （RA）, bone marrow stromal cells （BMSCs） can differentiate into nerve cells or neuron-like cells, which do not survive for a long time, so those are restricted to an application. Other neurotrophic factors can also differentiate into neuronal cells through inducing BMSCs; especially, brain-derived neurotrophic factor （BDNF） can delay natural death of neurons and play a key role in survival and growth of neurons. The combination of them is beneficial for differentiation of BMSCs. OBJECTIVE： To investigate the effects of BDNF combining with RA on inducing differentiation of BMSCs to nerve cells of adult rats and compare the results between common medium group and single BDNF group. DESIGN： Randomized controlled animal study SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College MATERIALS： The experiment was carried out in the Clinical Neurological Laboratory of Xuzhou Medical College from September 2003 to April 2005. A total of 24 SD rats, of either gender, 2 months old, weighing 130-150 g, were provided by Experimental Animal Center of Xuzhou Medical College [certification： SYXK （su） 2002-0038]. Materials and reagents： low-glucose DMEM medium, bovine serum, BDNF, RA, trypsin, separating medium of lymphocyte, monoclonal antibody of mouse-anti-nestin, neuro-specific enolase, glial fibrillary acidic protein （GFAP） antibody, SABC kit, and diaminobenzidine （DAB） color agent. All these mentioned above were mainly provided by SIGMA Company, GIBCO Company and Boshide Company. METHODS： Bone marrow of SD rats was selected for density gradient centrifugation. BMSCs were undertaken primary culture and subculture; and then, those cells were induced respectively in various mediums in total of 3 groups, including control group （primary culture）, BDNF group （20 μg/L BDNF） and BDNF＋RA group （20 μg/L BDNF plus 20 μg/L RA）. On the 3^rd and the 7^th days after induction, BMSCs were stained immunocytochemically

关键词： cell bone Synergistic effects of brain-derived neurotrophic factor and retinoic acid on inducing the differentiation of bone marrow stromal cells into neuron-like cells in adult rats in vitro BMSCs BDNF acid

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宋代城池筑防技术与思想考论

历史教学（下半月） 2019年第5期 52-58页

作者：刘永海贵州师范大学历史与政治学院

宋代城池筑防技术在沿袭前代的基础上,有很多改良和提升。《梦溪笔谈》《守城录》《武经总要》等著作,以实战为基础,进行了很多城池筑防技术的讨论.城墙防护思想方面,提出了针对不同类型城池的城墙结构见解,并有对城墙四角防御的思考与... 详细信息

宋代城池筑防技术在沿袭前代的基础上,有很多改良和提升。《梦溪笔谈》《守城录》《武经总要》等著作,以实战为基础,进行了很多城池筑防技术的讨论.城墙防护思想方面,提出了针对不同类型城池的城墙结构见解,并有对城墙四角防御的思考与改进,而且充分认识并发挥羊马墙在守战中的作用。射击设施的创新方面,一是平头女墙及移动女墙的创制,二是对传统马面的大胆改进。城门防御思想方面,提出了“城门不可依旧制”,“城门贵多不贵少,贵开不贵闭”的理念,创造了城楼的两层三重门形制,取消钓桥与悬门。

关键词：宋代城池筑城

论道教的重生思想——基于元代道教史籍的考察

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河南师范大学学报（哲学社会科学版） 2017年第5期44卷 87-92页

作者：刘永海贵州师范大学历史与政治学院贵州贵阳550025

道教是热爱生命的宗教,重生观念是道教思想的重要内容。元代道教史籍中蕴藏着许多重生思想,最具代表性者有三点:一是注重自身生命的思想。对个体生命的尊重,是道教立教的重要思想基础,并由此生发出反对肉体成仙,提倡灵魂成仙的观念。二... 详细信息

道教是热爱生命的宗教,重生观念是道教思想的重要内容。元代道教史籍中蕴藏着许多重生思想,最具代表性者有三点:一是注重自身生命的思想。对个体生命的尊重,是道教立教的重要思想基础,并由此生发出反对肉体成仙,提倡灵魂成仙的观念。二是尊重他人乃至整个人类生命的思想。道教不仅热衷"个人得道",而且追求"度人成仙",借助"天道好生",宣扬重生止杀。三是爱护和尊重一切动植物等生命体的思想。元代道教史籍鲜明提倡人和动、植物平等的观念,倡导保护自然,爱护动植物,救物放生。

关键词：元代道教史籍重生思想

Identification and Characterization of a Novel Thymus Aging Related Protein Rwdd1

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Cellular & Molecular Immunology 2008年第4期5卷 279-285页

作者： Ning Kang Lian Duan Long Tang Shirong liu Chunman Li yonghai Li Qingfeng liu Yu Hu Lianxian Cui Wei He Department of Immunology School of Basic Medicine Peking Union Medical College and Institute of Basic Medical Sciences Chinese Academy of Medical Sciences Beijing 100005 China

By using DDRT-PCR and EST segment ligation, a novel mouse thymus involution related gene Rwddl was identified. The reading frame encoded a protein of 243 amino acid residues which contained an RWD domain at the N terminus. Rwddl expression in thymus was decreased in aged and oxidatively stressed mice. It was found to be expressed extensively in thymocytes and thymic epithelial cells. The expression level of Rwddl could affect the transactivation activity of androgen receptor （AR） in transiently transfected thymic epithelial cells. However, no direct interaction could be detected by fluorescence resonance energy transfer （FRET） analysis. In conclusion, Rwddl is a thymus involution related protein that may indirectly affect AR signaling pathway. Cellular ＆ Molecular Immunology. 2008;5（4）：279-285.

关键词： Rwdd 1 thymus aging androgen receptor

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浅谈我国古代官私书目中的道教典籍

图书馆杂志 2005年第9期24卷 64-68页

作者：刘永海北京师范大学古籍研究所 100875

道教典籍数量庞大,我国历代官私书目中收录的大量道教典籍,从分类方法和著录范围乃至著录旨趣上,与道经目录都有很大不同;历代官私书目之间在诸多方面也存在差异。通过对古代官私书目的考察,可以更好地了解道教典籍的发展演变和流传情况。

关键词：道教典籍古代官私书目中国分类方法著录范围

由孝道崇拜到忠孝净明:许逊信仰历程及其特征论析

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河南师范大学学报（哲学社会科学版） 2020年第5期47卷 89-95页

作者：刘永海贵州师范大学历史与政治学院/贵州民间文献研究中心贵州贵阳550025

许逊崇拜是产生、流传于我国本土,根植于中华民族丰厚土壤中的民族信仰。早期许逊孝道崇拜的诞生以魏晋之际动荡社会及频繁的水患灾害为背景,植根于悠久的孝亲文化传统,带有鲜明的地域特征。唐及北宋时期的许逊崇拜,是孝道信仰与道教神... 详细信息

许逊崇拜是产生、流传于我国本土,根植于中华民族丰厚土壤中的民族信仰。早期许逊孝道崇拜的诞生以魏晋之际动荡社会及频繁的水患灾害为背景,植根于悠久的孝亲文化传统,带有鲜明的地域特征。唐及北宋时期的许逊崇拜,是孝道信仰与道教神学紧密结合的产物,又融入了浓厚的家族观念。南宋时期的高道大德,基于孝道信仰,借鉴儒释道心性学说,吸收日神、月神崇拜精华,首次以“净明”为名开宗立教,并借鉴儒学纲常伦理,在原来孝道信仰基础上,增益“忠”的内容。元代净明道依托传统孝道信仰,融合中华民族传统文化,构建了一套完整的宗教神学体系。同时进一步吸收儒学思想,将忠孝置于无以复加的位置,突出其在修道成仙过程中的特殊作用。明清净明道则呈现出与道教各派之融会交参以及向世俗化回归等特色。

关键词：净明道许逊信仰忠孝

Possible mechanism of reduced tiller number in the ascorbic acid-de ficient rice suppressed for L-galactono-1, 4-Lactone dehydrogenase

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Possible mechanism of reduced tiller number in the ascorbic ...

2016年全国植物生物学大会

作者： Le Yu yonghai liu Yezheng Chen Liping Zhou Hua Chen College of Life Sciences Zhaoqing University

The tiller of rice(Oryza sativa L.),which determines the panicle number per plant,is an important agronomic trait for grain production. Ascorbic acid(Asc) is a major plant antioxidant that serves many functions in plants. L-galactono-1,4-lactone dehydrogenase(GLDH,EC 1.***.2.3) is an enzyme that catalyzes the last step of Asc biosynthesis in plants. Here we show that the GLDH-suppressed transgenic rices,GI-1 and GI-2,which have constitutively low(between 30% and 50%) leaf Asc content compared with the wild-type plants,exhibit significantly reduced tiller number. Moreover,lower growth rate and plant height were observed in the Asc-deficient plants relative to the trait values of the wild-type plants at different tillering stages. Further examination showed that the deficiency of Asc resulted in a higher lipid peroxidation,a loss of chlorophyll,a loss of carotenoids and a lower rate of CO assimilation. In addition,the level of abscisic acid was higher in GI-1 plants,while the level of jasmonic acid was higher in GI-1 and GI-2 plants at different tillering stages. The results we presented here indicated that Asc deficiency was likely responsible for the promotion of premature senescence,which was accompanied by a marked decrease in photosynthesis. These observations support the conclusion that deficiency of Asc alters tiller number in the GLDH-suppressed transgenics through promoting premature senescence and changing phytohormones related to senescence.

关键词： ascorbic acid L-galactono-1 4-lactone dehydrogenase rice tiller number

来源： cnki会议评论

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道教仙传述略

图书与情报 2006年第4期 113-116,121页

作者：刘永海唐山师范学院政史系河北唐山063000

道教仙传自成系统,数量可观,其编撰历程可分为发轫期、成熟期、鼎盛期及衰贼期四个阶段;道教仙传的体裁可分为专传、类传、总传、像传四种。

关键词：道教仙传