Modification of Energy Metabolism-related Proteins is Responsible for Ceftriaxone-resistance

随着抗生素药物的开发与利用,细菌在对抗药物过程中逐渐发展出各种不同的耐药机制.近年来高通量的蛋白质组学技术已逐渐用于细菌耐药性机理研究,但主要集中在对细菌外膜蛋白的作用进行分析.本文采用2-d nATIVE/SdS PAgE方法从蛋白质复合物角度分析接近生理条件的胞浆蛋白在头孢曲松耐药性中的作用.结果发现8个耐药性相关蛋白质,通过对蛋白质功能分析,揭示了细菌通过调整能量代谢相关蛋白产生耐药性的新机制.进一步对相关菌株的次抑菌浓度和生存率分析,提示MAlP和SuCC等关键蛋白质可作为设计和开发新型抗菌分子的作用靶点.Outer membrane proteins of bacteria have been investigated systematically in response to antibiotic resistance by high-throughput proteomic techniques in recent years,but the data of the cytoplasmic proteins analyzed on a proteome scale are not available yet.In this study,2-D native/SDS-PAGE was first applied for characterization in cytoplasmic proteins responding to ceftriaxone-resistance in Escherichia coli.Eight proteins involved in modification of energy production and conversion were identified responsible for ceftriaxone-resistance.Among these proteins,down-regulated MalP and up-regulated SucC were found to play critical roles in response to resistance according to the MIC and survival ability analysis,and the two proteins may be candidate targets for development of novel antimicrobial drugs.浙江省自然科学基金项目(No.Y2090396);浙江理工大学科研启动基金项目(No.11615432610862)---

慈善捐赠对业绩异质性企业价值的影响研究——来自中国上市公司的经验证据

慈善捐赠作为企业履行社会责任的重要表现形式,受到了社会的广泛关注。现有关于慈善捐赠与企业价值之间关系的研究尚未得到一致的结论,单纯考虑二者关系可能使所得结论有失偏颇,本文将企业相对业绩纳入到分析框架,重点研究了慈善捐赠对企业价值的影响在不同业绩水平下是否有所差异。本文采用2003—2015年中国沪深两市A股上市公司为研究样本,研究了慈善捐赠对业绩异质性企业价值的影响。研究发现:相对于业绩较好的企业,业绩较差企业的慈善捐赠价值效应明显更弱,这表明绩差企业的捐赠行为更多的是代理问题的体现,而非利于企业发展的战略性行为;考虑市场化进程和政策不确定性的影响,该现象主要表现在市场化程度较低的地区和政策不确定性较高的地区。本文的研究有助于更好地认识慈善捐赠的经济后果,同时也为政府监管者监督绩差企业慈善捐赠行为提供了理论依据。中央高校基本科研业务费专项资金一般项目“宽松货币政策下制造业企业房地产投资对企业业绩的影响机理研究”(项目编号:XDJK2019C006)重庆市社会科学规划项目“非金融上市公司金融化的同伴效应研究”(项目编号:2018PY61)西南大学人文社会科学校级研究项目重大项目培育“新常态下正式与非正式制度性因素对公司亏损逆转稳定性的影响研究”(项目编号:SWU1909031

Changes in expression and localization of heat shock protein 70 during curcumin-induced apoptisis of human esophageal cancer cell line EC9706

目的探讨姜黄素对人食管癌EC9706细胞凋亡的诱导作用,热休克蛋白70(HSP70)在肿瘤细胞凋亡过程中在核基质上的变化及其与凋亡调控相关蛋白的关系。方法用细胞计数和流式细胞仪检测姜黄素对人食管癌EC9706细胞的增殖抑制作用,以光学显微镜和透射电镜观察姜黄素诱导人食管癌EC9706细胞凋亡前后的细胞结构变化,琼脂糖凝胶电泳观察人食管癌EC9706细胞凋亡前后的dnA结构变化。双向凝胶电泳和质谱鉴定分析HSP70在核基质中的存在与变化;并以WESTErn blOTTIng进行确证;激光扫描共焦显微镜观察HSP70在EC9706细胞凋亡过程中的定位及其与bAX、bCl-2等基因产物的共定位关系。结果姜黄素能显著抑制人食管癌EC9706细胞增殖并诱导人食管癌EC9706细胞凋亡,双向凝胶电泳、质谱鉴定和结果发现并证实,HSP70在姜黄素处理前后的EC9706细胞核基质蛋白中的存在及其表达下调变化。激光扫描共焦显微镜观察结果显示,HSP70在EC9706细胞凋亡过程中与bAX、bCl-2等基因产物具有共定位关系,且其共定位区域发生了变化。结论姜黄素对人食管癌EC9706细胞具有显著的凋亡诱导作用;HSP70作为一种新发现的核基质蛋白,在姜黄素诱导人食管癌EC9706凋亡过程中的表达与分布发生了显著变化。HSP70与凋亡相关基因的关系对EC9706细胞凋亡具有重要影响。Objective To investigate the effect of curcumin on human esophageal cancer EC9706 cells and explore the role of heat shock protein(HSP70) in cell apoptosis by examining changes in the nuclear matrix and its relationship with apoptosis-related proteins.Methods Cell counting and flow cytometry were performed to probe the inhibitory effect of curcumin on cellular proliferation.Transmission electron microscopy and optical microscopy were used to observe the structural changes in EC9706 cells before and after apoptosis.Agarose gel electrophoresis was conducted to investigate the DNA structure of EC9706 cells before and after apoptosis.Two-dimensional polyacrylamide gel electrophoresis(2-D PAGE) and mass spectrometry(MS) analysis were performed to investigate the presence and changes of HSP70 in the nuclear matrix of EC9706 cells before and after curcumin treatment,which was further corroborated by Western blotting assay.Laser confocal scanning microscopy was used to observe the colocalization of HSP70 with Bax and Bcl-2 during apoptosis.Results The results indicated that curcumin could markedly inhibited EC9706 cell proliferation and finally induced apoptosis.Data from 2-D PAGE,MS,and Western blotting showed that HSP70 was involved in the nuclear matrix proteins and expression of HSP70 was downregulated after curcumin treatment.Laser confocal microscopy showed that HSP70 colocalized with Bax and Bcl-2,and the colocalized regions were altered by the curcumin treatment.Conclusion Our work proves that curcumin could definitely induce EC9706 cells into apoptosis.As a new found nuclear matrix protein,the expression and distribution of HSP70 are altered during the apoptosis of EC9706 cells.The colocalization of HSP70 with apoptosis-related genes evidently affects the apoptosis of EC9706 cells.福建省自然科学基金资助项目(2011J01256); 中央高校基本科研业务费专项资金(2011121061

New Perspectives and Demonstration Research on Sport Cultural Exchanges between Fujian and Taiwan

概览两岸学者闽台体育文化交流学术研究成果,探讨两岸学者对闽台体育文化交流可持续性发展愿景,论述闽台体育文化交流的迫切性和可行性。通过多类、多项闽台体育文化交流项目的实验设计,实践反馈,实证研究,构建两岸体育文化交流可行的路径。通过对交流过程控制的剖析,对过程目标、形式、内容、方法等基本因素的系统整合,探索两岸体育文化交流合作面临的、急需解决的体制兼容、机构对接、条例配套等系列问题。提出闽台体育文化交流的研究重在创新和具有指导思想明确、项目计划严谨、措施科学合理、机制长效互动的新思路。This paper is to discuss the immediate need and feasibility of the sport cultural exchanges between Fujian and Taiwan in the light of the scholars' anticipation from both sides.In order to keep a constant development of the sport cultural exchanges between Fujian and Taiwan,through a thorough analysis on the experiences drawn from our past projects,we argue that the focus of the research on the exchanges should be placed on producing creative ideas with more well-designed projects which promote long-term interactions.2005年度国家社会科学基金项目(05BTY006

An Investigation of the Professional Development of Primary and Secondary School P. E. Backbone Teachers in Fujian Province

采用文献法、调查法、统计法、逻辑法等研究方法,以参加2013—2015年福建省基础教育万名骨干教师培训中的190名中小学体育骨干教师为调查对象,对教师基本情况和教师专业发展的认识、态度、知识、能力、继续教育等现状进行调查,在此基础上对教师专业发展现状进行分析,总结出我省中小学体育骨干教师专业发展的特点及存在的问题,并提出相应的建议,给我省中小学体育教师培训机构的改革提供依据,同时为我省中小学体育教师专业发展提供参考与借鉴,为进一步深化我省中小学体育教育改革提供人才保障.结果表明:1)目前,福建省中小学体育骨干教师以年富力强者居多,年龄、职称结构较合理;教师专业发展意识强,学历层次由大专逐步向本科、硕士过度,但硕士比例小;2)教师专业发展态度端正,职业的稳定性和责任感强烈,但部分年轻教师缺乏专业发展的自主性;3)教师对专业知识的掌握比较扎实,主要通过互联网、图书资料、专家报告或讲座等方法来获取专业知识;4)专业能力水平方面,教师在体育教学、课外指导、体育保健、现代教育技术运用等方面能力比较强,而创新教育和科研两方面能力相对比较薄弱;5)当下,我省中小学体育骨干教师最需要强化的培训是课题申报技能、论文写作能力、专题讲座能力、教育科研方法、提高学历层次、教学设计能力,其中提高科研能力和水平是教师们最迫切的愿望.2014年福建省中青年教师教育科研项目(社科A类)(JAS14023

Apoptosis of Human Esophageal Carcinoma Cell Line EC9706 Induced by Curcumin

目的:应用姜黄素处理人食管癌EC9706细胞,研究姜黄素对人食管癌EC9706细胞凋亡的诱导作用。方法:应用细胞计数、流式细胞仪、琼脂糖凝胶电泳、Hoechst染色、H.E染色和透射电镜检测经姜黄素诱导处理后人食管癌EC9706细胞的凋亡。结果:经姜黄素诱导处理后,人食管癌EC9706细胞生长抑制率达69.9%;细胞周期检测出现亚二倍体(亚G1期)细胞峰值,细胞凋亡率达23%;琼脂糖凝胶电泳显示出细胞凋亡典型的180-200 bp及其倍体的DNA"梯状"条带;Hoechst染色显示细胞核内出现浓染致密的固缩形态或颗粒状荧光;光镜和电镜下可见典型的细胞凋亡特征:细胞体积缩小,染色体凝集,可见有成群或单独存在的凋亡细胞,电镜下可见凋亡小体存在。结论:姜黄素能够有效诱导人食管癌EC9706细胞的凋亡,从而进一步为食管癌等恶性肿瘤疾病的治疗和凋亡机理的研究提供重要基础和科学依据.Objective: To study the apoptosis effects of the human esophageal carcinoma cell line EC9706 cells induced by curcumin(Cur). Methods: The apoptosis effects of EC9706 cells induced by curcumin were detected by cell count, flow cytometry analysis, light microscope and electron microscope. Results: After treated with curcumin, the proliferation of EC9706 cells was inhibited, and the inhibitory rate was 69.9%. The results of flow cytometry analysis showed that curcumin could induce the emergence of the phase of apoptosis, and the rate was 23%. Agarose gel electrophoresis revealed that cell DNA fragment exhibited characteristic "DNA ladder".Cell nucleus concentrated and appeared granular fluorescence by Hoechst33258 staining. Light microscope and electron microscope showed that the morphology of the cells treated with curcumin appeared shrinked, cell nucleus concerntrated, chromatin agglutinated, mitochondria swelled, and apoptosis body formed. Conclusion: This study suggested that curcumin could induce apoptosis of the human esophageal carcinoma cell line EC9706 effectively, and provided important foundation and research proofs to study more about the therapy of esophageal carcinoma, the malignant tumor and apoptosis mechanisms.国家自然科学基金项目(30470877

便携式拉曼光谱仪快速检测废水中残留有机溶剂

本文通过使用便携式拉曼光谱仪来快速检测废水中残留有机溶剂的种类和含量。首先利用便携式拉曼光谱仪建立标准拉曼谱图库,然后用便携式拉曼光谱仪检测废水样品并与标准谱图库比对,即可获得废水中含有的有机溶剂的种类和含量。该方法简便、灵敏、快速,对于实时监测废水的排放具有重要意义。国家自然科学基金(21522508);;浙江省重点研发计划(2017C03002,2018C03077

Expression and Localization of hnRNP A2/B1 during Differentiation of Human Osteosarcoma MG-63 Cells Induced by HMBA

背景与目的:核基质蛋白的差异表达与细胞癌变和增殖分化调控关系密切。本研究观察了hnRNP A2/B1在诱导分化处理前后人成骨肉瘤MG-63细胞核基质中的存在和分布,及其与Actin、Prohibitin的共定位关系。方法:选择性抽提经环六亚甲基双乙酰胺(hexamethylene bisacetamide,HMBA)诱导处理前后的MG-63细胞核基质,并运用双向电泳、质谱分析、蛋白质印记杂交、免疫荧光、激光共聚焦等技术检测hnRNP A2/B1在核基质中的表达与定位变化,及其与相关蛋白的共定位关系。结果:双向电泳及蛋白质印迹杂交结果证实了hnRNP A2/B1存在于MG-63细胞核基质蛋白组分中,并在HMBA处理后细胞核基质中表达下调;免疫荧光显微镜观察显示hnRNP A2/B1定位在核基质上,经HMBA处理后hnRNP A2/B1表达减弱。激光共聚焦显微镜观察结果显示,hnRNP A2/B1与细胞核基质蛋白组分Actin、细胞增殖相关调控因子Prohibitin具有共定位关系,但在诱导处理后细胞内的共定位关系减弱。结论:hnRNP A2/B1在MG-63细胞诱导分化过程中的表达分布,及其与Actin、Prohibitin的共定位关系的改变对MG-63细胞分化具有重要影响,值得进一步探索和研究。BACKGROUND & OBJECTIVE: The differentially expressed nuclear matrix proteins have great effects on canceration and regulation of cell differentiation. This study was to explore the existence and distribution of ribonucleoprotein hnRNP A2/B1 in nuclear matrix and its co-localization with Actin and Prohibitin in human osteosarcoma MG-63 cells before and after hexamethylene bisacetamide (HMBA) treatment. METHODS: The nuclear matrix of MG-63 cells before and after treatment of HMBA were selectively extracted. The expression and localization of hnRNP A2/B1 in nuclear matrix were detected by 2-D PAGE, MALDI-TOF-MS, Western blot, and immunofluorescent staining. The co-localization of hnRNP A2/B1 with Actin and Prohibitin was observed under laser scanning confocal microscope (LSCM). RESULTS: hnRNP A2/B1 was detected in the component of nuclear matrix proteins of MG-63 cells by Western blot and immunogold staining and its expression was decreased after treatment of HMBA. hnRNP A2/B1 was located in the nuclear matrix, and its expression was weakened after HMBA treatment. hnRNP A2/B1 was co-localized with Actin or Prohibitin in MG-63 cells, while the co-localization relationship was weakened during differentiation of MG-63 cells. CONCLUSIONS: hnRNP A2/B1 is a kind of nuclear matrix protein, and localizes in the nuclear matrix. The distribution and expression of hnRNP A2/B1 and its co-localization with Actin and Prohibitin play important roles during the differentiation of MG-63 cells.国家自然科学基金项目(No.30470877)~

基于内毒素血症探讨皂术茵陈方对非酒精性脂肪性肝炎大鼠的影响

目的 基于内毒素血症探讨皂术茵陈方防治非酒精性脂肪性肝炎(NASH)大鼠的影响及其作用机制。方法 采用随机数字表法将40只大鼠分为5组,即正常组、模型组、皂术茵陈方组、盐酸吡格列酮组及培菲康组,每组8只。除正常组外,其余32只大鼠采用高脂饮食16周建立大鼠NASH模型,在造模第9周开始,皂术茵陈方组给予皂术茵陈方水提液60 mg/(kg·d)灌胃,盐酸吡格列酮组给予盐酸吡格列酮10 mg/(kg·d)灌胃,培菲康组给予培菲康210 mg/(kg·d)灌胃,正常组及模型组均给予双蒸水10 mL/(kg·d)灌胃,共治疗8周。第16周末经腹主动脉取血,生化法检测肝组织甘油三酯(TG)水平,苏木精—伊红(HE)染色观察肝组织病理学变化。终点显色法检测血浆内毒素(LPS)含量。ELISA法检测肝组织肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6等炎症因子的表达水平。结果 与正常组比较,模型组大鼠肝组织显示出典型的NASH组织学特征,出现重度脂肪变性,不同程度的炎细胞浸润和坏死灶。大鼠肝湿重、肝指数、肝脏TG含量、血浆LPS水平、肝组织TNF-α、IL-1β、IL-6等炎症因..

中国式学科评估:问题与出路

今年四月份,教育部学位与研究生教育发展中心(以下简称"教育部学位中心")邀请全国学位授予单位参加全国第四轮一级学科整体水平评估。随之,各个高校展开了一场大规模、高级别的学科评估申报及材料提交总动员。第四轮学科评估自发布起也引发了学界的广泛关注和热烈讨论。高等教育是中国崛起的思想发动机,关涉民族复兴的未来,而学科评估是近年来中国高等教育学科建设成就的集中展示,其意义和影响可谓深远。为了更好推进学科评估科学进行,特别是促进高等教育健康发展,《探索与争鸣》编辑部邀请全国