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Cloning and Analysis of the Promoter Region of Rat uPA Gene

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Journal of Reproduction and Contraception 2007年第1期18卷 1-10页

作者： Yan LIU Jin-wen XIONG Li-gang CHEN yong-hong tian Cheng-liang XIONG i Medical College HuazhongUniversity of Science and Technology Wuhan 430030China

Objective To clone and analyze the promoter sequence of rat urokinase plasminogen activator protein gene. Methods The genomic DNA was extracted from rat testicular tissue. According to urokinase plasminogen activator, the gene sense primer and antisense primer of uPA gene were designed and synthesized, then Touch-Down PCR were performed. After proper purification, the PCR product was sequenced, analyzed with the promoter prediction software and compared with the DNA sequence of rattuas urokinase plasminogen activator. Results The cloned uPA gene was about 1 572 bp in length, which contained a full open-reading frame with 21 bp in length exons, and the upper region of transcriptional start was 1 551 bp in length which was eucaryon transcriptional control area. The 5＇ UTR had a promoter region including a non-responsive TATA-box. Not only the GC-box binding region was found in this gene, but also active protein I （AP1） and SP1 were seen in other regions. Conclusion A 1 572 bp uPA gene fragment （GenBank accession No.X65651） was obtained from rat genomic DNA library, containing eucaryon transcriptional control area with a promoter region, non-conspicuous TATA-box, GC-box and an extron. A non-responsive TATA-box is located at the upper -30 region.

关键词： rat testicular tissue urokinase plasminogen activator Touch-Down PCR promoter region eucaryou transcriptional control area

Gene expression changes of urokinase plasminogen activator and urokinase receptor in rat testes at postnatal stages

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Asian Journal of Andrology 2007年第5期9卷 679-683页

作者： Dong-Hui Huang Hu Zhao yong-hong tian hong-Gang Li Xiao-Fang Ding Cheng-Liang Xiong Family Planning Research Institute Tongji Medical College Huazhong University of Science ＆ Technology Wuhan 43003 China Department of Human Anatomy Tongji Medical College Huazhong University of Science ＆ Technology Wuhan 43003 China

Aim： To investigate the gene expression changes of urokinase plasminogen activator （uPA）/urokinase receptor （uPAR） in rat testes at postnatal stages and explore the effects of uPA/uPAR system on the rat spermatogenesis. Methods： The mRNAs of uPA and uPAR in rat testes were measured by using real-time quantitative polymerase chain reaction （PCR） at postnatal days 0, 5, 10, 15, 21, 28, 35, 42, 49 and 56, respectively. Results： The tendencies of uPA and uPAR mRNA expression were similar at most postnatal stages except for Do. The expression of uPAR mRNA in rats testes was relatively higher than that of uPA at postnatal Do, and both were decreased until D21, increased obviously at postnatal D28, reached a peak at postnatal D35, then declined sharply at postnatal D42 and retained at a low level afterwards. Conclusion： The uPA/uPAR system may be strongly linked to spermiation and spermatogenesis via regulating germ cell migration and proliferation, as well as promoting the spermiation and detached residual bodies from the mature spermatids.

关键词： rats spermatogenesis urokinase plasminogen activator urokinase receptor quantitative polymerase chain reaction

Towards human-like and transhuman perception in AI 2.0:a review

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Frontiers of Information Technology & Electronic Engineering 2017年第1期18卷 58-67页

作者： yong-hong tian Xi-lin CHEN hong-kai XIONG hong-liang LI Li-rong DAI Jing CHEN Jun-liang XING Xi-hong WU Wei-min HU Yu HU Tie-jun HUANG Wen GAO School of Electronics Engineering and Computer Science Peking University Institute of Computing Technology Chinese Academy of Sciences Department of Electronic Engineering Shanghai Jiao Tong University School of Electronic Engineering University of Electronic Science and Technology of China Department of Electronic Engineering and Information Sciences University of Science and Technology of China School of Optoelectronics Beijing Institute of Technology Institute of Automation Chinese Academy of Sciences

Perception is the interaction interface between an intelligent system and the real world. Without sophisticated and flexible perceptual capabilities, it is impossible to create advanced artificial intelligence （AI） systems. For the next-generation AI, called ＇AI 2.0＇, one of the most significant features will be that AI is empowered with intelligent perceptual capabilities, which can simulate human brain＇s mechanisms and are likely to surpass human brain in terms of performance. In this paper, we briefly review the state-of-the-art advances across different areas of perception, including visual perception, auditory perception, speech perception, and perceptual information processing and learning engines. On this basis, we envision several R＆D trends in intelligent perception for the forthcoming era of AI 2.0, including：（1） human-like and transhuman active vision; （2） auditory perception and computation in an actual auditory setting; （3） speech perception and computation in a natural interaction setting; （4） autonomous learning of perceptual information; （5） large-scale perceptual information processing and learning platforms; and （6） urban omnidirectional intelligent perception and reasoning engines. We believe these research directions should be highlighted in the future plans for AI 2.0.

关键词： Intelligent perception Active vision Auditory perception Speech perception Autonomous learning

Role of Urokinase-type Plasminogen Activator in the Precontact Sperm-egg Communication and Fertility of Mice in vitro

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Journal of Reproduction and Contraception 2005年第4期16卷 201-212页

作者： Xiao-fang DING Cheng-liang XIONG hong-gang LI yong-hong tian Jin-wen XIONG Lian HU Center of Reproductive Medicine Tongji Medical College Huazhong University of Science and Technologe Wuhan 430030 China

Objective To explore the role of urokinase-type plasminogen activator（uPA） in precontact sperm-egg communication and fertility of mice in vitro. Methods Firstly, sperm chemotaxis （SC） induced by uPA was assayed by measuring the sperm densities in capillaries with a descending gradient or no gradient of uPA respectively. Secondly, the role of uPAR that exists in sperm plasma membrane in SC was studied by examining the change of sperm density in capillary after incubating spermatozoa with anti-uPAR antibody. Thirdly, SC induced by eggs, which had been treated with uPA, PAl-1 and anti-uPAR beforehand respectively, was assayed to study the role of uPA in PSEC. Lastly, the fertilization capability of spermatozoa treated with uPA was examined by counting the number of fertilized eggs. Results 1）The density of spermatozoa that migrated down the gradient of uPA into the capillary was significantly lower than that into the capillary containing no-gradient uPA. 2） When uPAR of spermatozoa was inhibited by anti-uPAR antibody, the density of spermatozoa that migrated into the capillary with ascending gradient of uPA decreased correspondingly. 3） The density of spermatozoa attracted by eggs, which were treated with uPA beforehand, increased significantly than that of attracted by non-treated eggs. On the contrary, the sperm density decreased correspondingly when the egg was treated with PAI-1. 4） The number of fertilized eggs increased significantly after the spermatozoa used here was treated with uPA beforehand. Conclusion uPA could induce SC of mice sperm in vitro through the uPAR on its membrane, enhance the capability of egg inducing SC, and promote spermatozoa to fertilize eggs. Thus, uPA may act as an attractant in PSEC, increase the chance encounter of spermatozoa and eggs, therefore, enhance the fertility success correspondingly. This study, in some degree, provides an evidence that uPA may be used as a new medicine and diagnostic reagent for male infertility.

关键词： urokinase-type plasminogen activator sperm chemotaxis precontact sperm-egg communication fertility

Enhancing m^(6)A modification in the motor cortex facilitates corticospinal tract remodeling after spinal cord injury

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Neural Regeneration Research 2025年第6期20卷 1749-1763页

作者： tian Qin Yuxin Jin Yiming Qin Feifei Yuan hongbin Lu Jianzhong Hu yong Cao Chengjun Li Department of Spine Surgery and Orthopedics Xiangya HospitalCentral South UniversityChangshaHunan ProvinceChina Key Laboratory of Organ Injury Aging and Regenerative Medicine of Hunan ProvinceChangshaHunan ProvinceChina National Clinical Research Center for Geriatric Disorders Xiangya HospitalCentral South UniversityChangshaHunan ProvinceChina Department of Sports Medicine Xiangya HospitalCentral South UniversityChangshaHunan ProvinceChina

Spinal cord injury typically causes corticospinal tract disruption. Although the disrupted corticospinal tract can self-regenerate to a certain degree, the underlying mechanism of this process is still unclear. N6-methyladenosine(m^(6)A) modifications are the most common form of epigenetic regulation at the RNA level and play an essential role in biological processes. However, whether m^(6)A modifications participate in corticospinal tract regeneration after spinal cord injury remains unknown. We found that expression of methyltransferase 14 protein(METTL14) in the locomotor cortex was high after spinal cord injury and accompanied by elevated m^(6)A levels. Knockdown of Mettl14 in the locomotor cortex was not favorable for corticospinal tract regeneration and neurological recovery after spinal cord injury. Through bioinformatics analysis and methylated RNA immunoprecipitation-quantitative polymerase chain reaction, we found that METTL14 regulated Trib2 expression in an m^(6)A-regulated manner, thereby activating the mitogen-activated protein kinase pathway and promoting corticospinal tract regeneration. Finally, we administered syringin, a stabilizer of METTL14, using molecular docking. Results confirmed that syringin can promote corticospinal tract regeneration and facilitate neurological recovery by stabilizing METTL14. Findings from this study reveal that m^(6)A modification is involved in the regulation of corticospinal tract regeneration after spinal cord injury.

关键词： corticospinal tract remodeling epigenetic regulations locomotor cortex m^(6)A modification methyltransferase 14 protein(METTL14) mitogen-activated protein kinase neural regeneration spinal cord injury syringin TRIB2

尿激酶型纤溶酶原激活因子在小鼠精-卵接触前通讯和体外受精中作用初探

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生殖与避孕 2006年第1期26卷 3-9页

作者：丁晓芳田永红李红钢熊锦文熊承良华中科技大学同济医学院计划生育研究所/生殖医学中心武汉430030

目的:探讨尿激酶型纤溶酶原激活因子(uPA)在雄性促生育中的作用机制。方法:通过检测精子在毛细管内聚集数量的变化来测试小鼠精子体外对uPA和卵细胞的趋化性,以及将卵细胞分别经uPA、uPA抑制剂(PAI-1)以及抗-uPAR抗体预处理后,检测其趋... 详细信息

目的:探讨尿激酶型纤溶酶原激活因子(uPA)在雄性促生育中的作用机制。方法:通过检测精子在毛细管内聚集数量的变化来测试小鼠精子体外对uPA和卵细胞的趋化性,以及将卵细胞分别经uPA、uPA抑制剂(PAI-1)以及抗-uPAR抗体预处理后,检测其趋化诱导精子的能力,来反应uPA在精-卵接触前通讯中的作用。其次观察精子经uPA预处理后其受精能力的变化。结果:精子在递减uPA浓度梯度中的聚集数量明显低于等浓度内的(P<0.05);经uPA预处理的卵细胞其聚集精子的能力显著强于未经处理的卵细胞(P<0.05),而用PAI-1预处理卵细胞后,卵细胞聚集精子的能力显著下降(P<0.05);用抗-uPAR抗体预处理卵细胞并不影响卵细胞对精子的聚集作用;uPA处理后的精子与卵细胞共孵育后其受精卵数目多于未经处理的(P<0.05)。结论:精子在体外对uPA和卵细胞均有趋化性;uPA能增加卵细胞对精子的趋化作用,增进精-卵接触前通讯,促进受精。本实验在一定程度上探讨了uPA促男性生育的机制,并为其在临床上的应用提供一定的理论依据。

关键词：尿激酶型纤溶酶原激活因子(uPA) 精-卵接触前通讯精子趋化性体外受精小鼠

大鼠睾丸组织uPA基因启动子区的克隆与分析

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生殖与避孕 2006年第9期26卷 520-525页

作者：刘燕熊锦文陈丽刚田永红熊承良华中科技大学同济医学院计划生育研究所生殖医学中心武汉430030

目的:克隆大鼠睾丸组织uPA基因的启动子区并对该序列进行分析。方法:从大鼠睾丸组织中提取基因组DNA,以其为模板设计uPA基因引物,运用降落PCR法扩增uPA基因5'端上游的真核转录调控序列。测序得到的PCR产物用启动子区的分析软件分析,并与... 详细信息

目的:克隆大鼠睾丸组织uPA基因的启动子区并对该序列进行分析。方法:从大鼠睾丸组织中提取基因组DNA,以其为模板设计uPA基因引物,运用降落PCR法扩增uPA基因5'端上游的真核转录调控序列。测序得到的PCR产物用启动子区的分析软件分析,并与其DNA序列进行比对。结果:得到的uPA基因长度为1572bp(登录号X65651)。经软件分析:该序列包含uPA基因完整的开放阅读框(ORF)、21bp的外显子部分,1551bp区域为转录起始的上游部分在其5'端UTR区的-30bp位置有一个不典型的TATA盒,其上游有非常明显的GC盒及启动子区常见的AP1(activeprotein1)、SP1等结合位点。结论:成功克隆了大鼠睾丸组织uPA基因启动子区。分析表明,该片段包含真核转录调控区域、不典型的TATA盒、典型的GC盒及启动子区所常见的AP1、SP1等结合区域。

关键词：大鼠睾丸组织尿激酶纤溶酶原激活因子(uPA) Touch-Down PCR 启动子区真核转录调控区域