度甘氏矩阵图研究会周年庆暨

甘氏矩阵图Q＆A壹、价格篇一、认识矩阵图1. Square of Four 与 Square of Nine2. 矩阵图与轮中轮二、基础看图法1.大数，百分比运用2. 基础看图法：角线对角线‧十字对十字3. 如何看一圆周4. 角度线区域判别5. 角线、十字线撑压问题6. 如何观察辅线动能7. 个股推算要采取还原权值股价还是原始股价三、切线、暂时基数1. 短切线2. 短切线、中心点位与百分率1/4停利法/font>四、矩阵小图1.小图五、进场与出场1. 一角停损法2. 角度区间进场法短切线3. 角度区间停利法4. 该如何停利贰、时间篇1. 时间与价位关系2. 时间、生日推算3. 变盘时间参、观念与操作篇1. 技术分析自毁论2. 矩阵图的正确使用方式3. 量是不是不重要？4. 观察现货，操作期货=============================================================================== 「Square of Four」与「Square of Nine」2003-06-29 23:28 -Bucks：发现个小问题刚才在贴大图的时候，发现怎么叫做「Square of Four」，而不是「Square of Nine」呢?2003-06-30 23:21 -矩阵：bucks兄‧您的疑问真是好，有观察力！图送出去500，您是第一个提出来的，为什么是「Square of Four」，而不是「Square of Nine」呢？您有答案吗？当然，说对了有礼物。

有大图的人有兴趣，也可以“猜猜”看！2003-07-01 08:03 -OKLA：猜猜看：我猜是因为它本来就是四角形，而且以十字分割成四个象限，也都是四角形。

也就是说以十字分割成四个四角形的组合！ (Square of Four!)2003-07-24 08:15 -djsl：我的看法稍有不同不是十字线将矩阵（形）切割成四个区块，而是两条对角线将矩阵（形）切割成四个区块。

影响网购冲动消费行为因素分析——以双十一为例目录1绪论 (1)1.1研究背景 (1)1.2研究目的及意义 (1)1.3研究方法 (2)1.4研究内容 (2)1.5研究贡献度 (2)2文献综述 (3)2.1冲动性购买行为 (3)2.2冲动购买意愿 (5)2.3情感反应 (5)2.4网站特性 (6)2.5打折促销手段 (6)2.6个人冲动特质 (7)3研究方法 (7)3.1研究假设 (7)3.2研究框架 (9)3.3问卷设计 (10)3.4样本的选择与容量 (12)3.5问卷试做 (12)4研究结果与分析 (12)4.1样本与回收率 (12)4.2数据分析结果 (12)5讨论与启示 (19)5.1研究总结 (19)5.2研究局限 (21)5.3对未来研究的建议 (21)参考文献 (22)[论文摘要]本文以双十一网络狂欢购物节为研究对象，目的是探讨影响网购冲动消费行为的因素，通过构建回归模型，探讨网站特性（包括网站知识性、视觉性、互动性）、打折促销手段和消费者个人冲动特质与消费者积极情感反应的关系，积极情感反应与冲动性购买意愿的关系，冲动性购买意愿与实际冲动性购买行为的关系，通过随抽样法进行问卷发放，回收问卷数据对理论模型进行实证研究，运用IBM SPSS Statistics 22对回收的403份有效调查问卷数据进行信度分析、回归模型分析，得出除网站知识性对积极情绪反应无显著性外，其他因素对于能够引发冲动购买意愿的积极情绪反应都有正向影响，从而为参与双十一的商家和消费者提出建议。

[关键词]网站特性；打折促销手段；个人冲动特质；积极情绪反应；冲动购买[Abstract] The paper takes the Double Eleven which is an online shopping carnival as the research object, the purpose of which is to explore the factors affecting impulsive consumption behavior of online shopping. By constructing a regression model, exploring the relationship between the characteristics of the website (including the knowledge, visuality, interactivity of the website), the means of discount and promotion, consumers' personal impulsive characteristics and consumers' positive emotional reactions, the relationship between positive emotional reactions and impulsive purchase intentions, and the relationship between impulsive purchase intentions and actual impulsive purchase behaviors, distributing the questionnaire by the random sampling method and then collecting the questionnaire data to conduct empirical research on the theoretical model, this paper conducts reliability analysis and regression model analysis on the valid data of 403 questionnaires by using IBM SPSS Statistics 22. It is concluded that in addition to the fact that the knowledge of the website is not significant in responding to positive emotional reactions, other factors have a positive impact on the positive emotional reactions that can trigger the impulsive purchase intentions, thus making recommendations for merchants and consumers participating in the Double Eleven.[Key words] website characteristics; discount promotion means; personal impulse trait; positive emotional response; impulse purchase1 绪论1.1 研究背景在网络信息技术日新月异的现代，网络经济作为不容忽视的经济形式，而且不得不说网络购物已经成为网络经济的重要表现形式，随着网络的普及和智能手机的发展，越来越多人加入到网络购物的潮流中来。

簡介進入本書之前，您要先知道，您不會在這一本書中找到只贏不輸的方法，也沒有次次能測底摸頭的神算技巧，是的，沒有一勞永逸、一夕致富這樣的事。

想要改變輸錢劣境？想要學會得心應手的操作技能？想要重回初入市場時，對未來人生之路所築構的夢？都不是難事！首要丟棄之前的固執心態，與錯誤觀念，再放下過去的痛苦經驗，放下吧！重新開始建立截然不同的操作基礎。

您選擇了矩陣圖作為新生命的開始，很好，矩陣圖自是不會讓您失望。

除了改變觀念，重建操作心態，您要做比以往更多的功課，做更多的比對，要更認真與努力，且堅持，只要準備好，我們就能擁抱財富。

前言從價格篇出版後，這些年從讀者的建言中，從許多問題的討論經驗中得知，剛開始研讀這本書時，其實不是很輕鬆，但一旦進入狀況，其實並不難懂。

於是，時間篇的開始，要先提出建議，請以輕鬆的心情進入您的時間之旅，但是不要如看小說般隨意翻閱，不用急，給自己多些閱讀的時間，看完一遍之後，第二遍再來逐步探討，理解之後，才作更進一步的深入學習。

如對學習進度仍有疑問，請先翻看，第五章的『價格篇的學習順序』、『時間篇的學習順序』。

我們從價格篇開始，即強調學習矩陣圖要『空杯進入』。

除了空杯才能再裝進新的東西之外，重點在，矩陣圖是數字的表現應用，而一般圖表分析是曲線的表現，雖然都是技術分析，最終的目的都是一樣，但是學習的觀念及方式是全然不同的，如果在學習過程中思考及應用時加入其它的東西，是不會讓學習的速度加快，讓理解的範圍加大，基本上不會有加分的效果，反而會產生阻礙。

矩陣圖中包含了許多東西，只要簡單使用，即會產生最大的力量，加什麼東西進去都是畫蛇添足，請學習者一定要先認知這一點。

進入學習後，接著請要『按部就班』的走，一定是瞭解一個章節的說明或推算使用後，才進入下一段。

因為矩陣圖的推算技巧，每一種都能單獨運作，不是要從頭到尾的熟悉後才能運用。

按進度推進，一路上所有的推算技巧都會有一個學習、熟練的空間，一個一個先後的熟悉運用後，學習者將可以輕易的將它們依短、中線的推算需求去單獨使用，或組合使用，這才是真正會使用矩陣圖了。

甘氏矩阵图Q＆A壹、价格篇一、认识矩阵图1. Square of Four 与 Square of Nine2. 矩阵图与轮中轮二、基础看图法1.大数，百分比运用2. 基础看图法：角线对角线‧十字对十字3. 如何看一圆周4. 角度线区域判别5. 角线、十字线撑压问题6. 如何观察辅线动能7. 个股推算要采取还原权值股价还是原始股价三、切线、暂时基数1. 短切线2. 短切线、中心点位与百分率1/4停利法/font>四、矩阵小图1.小图五、进场与出场1. 一角停损法2. 角度区间进场法短切线3. 角度区间停利法4. 该如何停利贰、时间篇1. 时间与价位关系2. 时间、生日推算3. 变盘时间参、观念与操作篇1. 技术分析自毁论2. 矩阵图的正确使用方式3. 量是不是不重要？4. 观察现货，操作期货=============================================================================== 「Square of Four」与「Square of Nine」2003-06-29 23:28 -Bucks：发现个小问题刚才在贴大图的时候，发现怎么叫做「Square of Four」，而不是「Square of Nine」呢?2003-06-30 23:21 -矩阵：bucks兄‧您的疑问真是好，有观察力！图送出去500张，您是第一个提出来的，为什么是「Square of Four」，而不是「Square of Nine」呢？您有答案吗？当然，说对了有礼物。

有大图的人有兴趣，也可以“猜猜”看！2003-07-01 08:03 -OKLA：猜猜看：我猜是因为它本来就是四角形，而且以十字分割成四个象限，也都是四角形。

也就是说以十字分割成四个四角形的组合！ (Square of Four!)2003-07-24 08:15 -djsl：我的看法稍有不同不是十字线将矩阵（正方形）切割成四个区块，而是两条对角线将矩阵（正方形）切割成四个区块。

甘氏矩阵图跑图程式Gann-9+好风光好风光恢复供货才甘氏矩陣時間價格跑圖程式+Gann-9使用說明手冊23寫在前面 ..................................................................... .............................. 7 第一次使用程式: .................................................................... ................. 7 程式的更新與啟動 ..................................................................... ............. 8 簡介 .......................................................................................................... 9 如何使用 ..................................................................... .............................. 9 功能說明 ..................................................................... .............................. 7 檔案 ..................................................................... (7)開啟新檔.. ................................................................... .. (7)開啟舊檔 ..................................................................... (11)儲存檔案 ..................................................................... (13)另存新檔 ..................................................................... (10)另存圖檔 ..................................................................... (14)列印 ..................................................................... . (14)預覽列印 ..................................................................... (14)列印設定 ..................................................................... (14)結束 ..................................................................... ............................ 14 工具列 ..................................................................... . (15)4標準模式 ..................................................................... ..................... 15 標注顏色 ..................................................................... ..................... 15 產生切線 ..................................................................... ..................... 15 產生短切線 ..................................................................... .................. 16 大數對應 ..................................................................... ..................... 16 畫線工具 ..................................................................... ..................... 16 基礎角度線 ..................................................................... .................. 18 角線區域 ..................................................................... ..................... 18 工具(選單) .................................................................... ....................... 18 快速製圖機.. ................................................................... .................. 18 快速搜尋.. ................................................................... ..................... 19 檢視 ..................................................................... ............................... 19 基礎角度線.. ................................................................... .................. 19 角線區域.. ................................................................... ..................... 19 放大.. ................................................................... ............................ 20 縮小.. ................................................................... ............................ 20 工具列 ..................................................................... ........................ 20 狀態列 ..................................................................... ........................ 21 說明 ..................................................................... . (21)5甘氏矩陣圖研究會 ..................................................................... ........ 21 註冊&下載中心 ..................................................................... (21)+關於Gann-9 ...................................................................... . (22)甘氏矩陣原圖 (18)甘氏矩陣日時間圖 (19)甘氏矩陣日時間價位圖 (20)甘氏矩陣月時間圖 (21)甘氏矩陣時間標準圖 (22)6寫在前面第一次使用程式:特別說明:當您第一次啟動程式時,將會自動前往註冊中心,在您依指示輸入資料後(*注意*請確認您資料的完整性,日後將依您所輸入的資料來進行驗証的動作。

甘氏时间标准图及矩阵图甘氏时间标准图（Gantt Chart）是一种以条状图形式表示项目进度计划的工具，也被称为甘特图或甘特图表。

它以水平条状图的形式将任务的起始日期、持续时间以及完成日期显示出来，使项目管理人员可以一目了然地查看整个项目的进度安排。

甘氏时间标准图主要由以下三个部分组成：1. 横坐标：用来表示时间轴，通常以天、周或月为单位。

时间轴的左侧用来标注任务的名称。

2. 纵坐标：用来表示参与项目的不同团队成员或资源，可根据需要进行分类或分组。

3. 条形图：条状图以横向显示任务的时间起点和时间跨度，并用不同的颜色或图案来表示任务的不同阶段或进度。

甘氏时间标准图可以为项目管理者提供以下几个方面的信息：1. 任务的起始日期和完成日期：通过条形图的长度和位置，可以很容易地了解各个任务的时间安排。

2. 任务的持续时间：条形图的长度代表任务的持续时间，可以帮助项目管理者预估每个任务所需要的时间。

3. 任务的依赖关系：通过将不同任务的条形图放置在正确的位置，可以显示任务之间的前后关系和依赖关系。

4. 任务的进度：项目管理者可以通过在条形图上标记实际完成日期来了解项目的实际进度，进而与计划进度进行比较。

甘氏时间标准图可以帮助项目管理者有效地规划、监控和控制项目进度。

通过将项目中的所有任务整齐地排列在一个图表上，项目管理者可以随时了解项目的整体进展情况，及时发现任务之间的冲突或瓶颈，并通过对任务的重新排列或分配资源来调整项目进度，保证项目能够按时完成。

除了甘氏时间标准图外，还有一种与之类似的工具叫做甘氏时间矩阵图（Gantt Matrix）。

甘氏时间矩阵图通过将任务的起始日期和完成日期以矩阵的形式呈现，更加直观地展示不同任务之间的时间关系。

甘氏时间矩阵图的主要特点如下：1. 使用方便：与甘氏时间标准图相比，甘氏时间矩阵图更加简洁明了，对于小型项目来说更容易理解和应用。

2. 显示任务之间的时间关系：矩阵图将任务的起始日期和完成日期以矩阵的形式呈现，更能突出任务之间的时间关系，便于项目成员协调工作。

甘氏矩陣圖價格推算『矩陣圖』是甘氏理論中最簡易明確的技術分析，是甘氏一直致力使用的推算工具，堪稱是甘氏一生的精華，甘氏理論的精髓。

一張四方形的數字圖，其中竟隱含了所有金融商品的進退趨勢 (當然也包括股票) ，此圖可以說，可推算所有有關數字的東西。

美國小說家，索爾‧貝婁說：『老朽的生命疲竭，而新的形式尚未出現，人們彷彿在沙漠逡巡探勘，努力找尋新的形式。

』這句話用於現在的技術分析領域裡，顯得非常貼切。

行情的變化往往於瞬息間，很多投資人每天都在猜猜看，當然，猜對了有獎！可是有多少人猜中大獎？又有多少人能連續猜對？任何有心於市場的投資人都知道，要想存活於市場絕非問問明牌、看看上市公司相關財務報表數據，或簡單學個技術分析、畫畫線、瞄瞄指標就可以掌握行情，可以傲視且屹立於市場！如果這麼簡單人人都賺翻了，也不會猜對十次，還不夠錯一次大的。

有一句話很好：『市場的錢贏不完，口袋裡的錢卻輸得完。

』長久以來技術分析工作者、投資大眾，不論新手老手，一直在找尋一種簡單明白、快速有效的分析工具，卻愈迷失在多如牛毛的技術指標、深奧的理論與複雜的線形中。

尤其拜科技之賜，現在的圖形分析幾乎全在電腦上進行。

電腦圖之速度，自是手工繪圖所望塵莫及的，但電腦圖常見的盲點在於只能參考過去的線形，無法切劃未來較長的行進方向 (邊度不夠) ；此外受限於電腦硬體的規格，或因軟體程式功能的限制與壓縮圖形往往失真及變形，並不能做為推算依據。

以一般技術分析軟體上常使用的甘氏角為例，根本形同虛設。

因為：『甘氏角必需架立在矩形圖上才能使用，現在的電腦圖都是略長形較多，更別說這套軟體的程式，也要是專為架設甘氏角寫的，且使用甘氏獨特的線形圖。

』現在大部份技術分析軟體都不符合這樣的條件，是無法作用的。

許多人不知道也用得很有意思（等有了問題才奇怪甘氏角不好用？），這是盤後技術分析軟體該檢討的問題，也是使用者應知道的事。

其實，每一種技術分析的指標及理論，都有其學習與參考的價值，問題是絕大多數都無法提供我們對未來行情變化的肯定。

#红柳⼈物#他是中国防震减灾领域的专家，在地震、泥⽯流的灾害现场，他都要下到坑洞或者地下室查看建筑物的隔震板状况，他对同事说，⾃⼰亲⾃做的检查才能放⼼，不能为了⾃⼰的安危不顾众多灾区百姓的安危。

他是2015年“全国师德标兵”，在课堂、实验室、学⽣宿舍，你都能看到他和学⽣交流的⾝影。

他对妻⼦说，我们的孩⼦，我不在的时候还有你，可学校⾥，⼏⼗个孩⼦在等着我。

他发起创建了⼀个名为“建⼯七七”的基⾦，作为兰州理⼯⼤学七七级校友的⼀员，他和他的同学们为了给母校学弟学妹更多的⿎励和资助，积极筹资，改⾰增值。

他对同学们说，要好好珍惜来之不易的机会，对得起校友，对得起母校。

他就是杜永峰，兰州理⼯⼤学⼟⽊学院教授，防灾减灾研究所主任，⼏⼗年如⼀⽇，他⽤发⾃内⼼的爱与奉献谱写了⼀⾸动⼈的育⼈之歌。

哪⾥有地震哪⾥就有他2008年汶川⼤地震，⽢肃省陇南市因为毗邻四川，⼀些地⽅也成为重灾区。

作为省内专家的杜永峰⽴即赶赴灾区，当他站在3栋⼏乎完好⽆损的住宅楼前时，“悬着的⼼⼀下⼦就落进了肚⼦⾥。

”这些六层楼的建筑，因为使⽤了他研究推⼴的“隔震垫”技术，墙体没有出现裂缝，住户家中的东西也没有因为地震发⽣掉落，这是⽢肃省内应⽤这⼀技术的建筑⾸次成功经受住地震的考验。

⽽这距离杜永峰的研究，已经过去了12年。

1996年，结构⼯程专业的杜永峰开始研究“隔震垫”技术。

这项起源于新西兰，最初在国内南⽅应⽤的技术对于地震的作⽤可谓和地基⼀样重要。

“隔震垫”的专业名称叫做“叠层橡胶隔震⽀座”，是⼀种由橡胶和钢板制作⽽成的圆柱形装置。

这种被放在建筑物地基与上体结构中间的⼩垫⼦，在地震来临时，起到了⾮常⼤的缓冲作⽤，上体结构的晃动会明显减⼩。

杜永峰不⼤的办公室⾥，堆满了研究⽤的设备、仪器、军⼤⾐，还有这近20年来他研究的各种“隔震垫”。

南北⽅的⽓候差异，导致“隔震垫”要进⼊北⽅，⾸先要适应这⾥的温度。

研究中，杜永峰带着他的学⽣们经常待在零摄⽒度以下的实验室⾥，把放置在零下50摄⽒度冷冻机⾥的“隔震垫”快速取出安装，然后接受模拟地震震动，虽然每次都穿着军⼤⾐，但他们⼀个个还是冻得直打哆嗦。

Original ContributionThe extracellular matrix metalloproteinase inducer EMMPRIN is a target of nitric oxide in myocardial ischemia/reperfusionCarlos Tarin a ,1,Begoña Lavin a ,1,Monica Gomez a ,Marta Saura b ,Antonio Diez-Juan c ,Carlos Zaragoza a ,⁎a Fundación Centro Nacional de Investigaciones Cardiovasculares,Madrid 28029,Spainb Departamento de Fisiologia,Facultad de Medicina,Universidad de Alcala,Madrid 28871,SpaincVascular Repair and Regeneration Laboratory,Department of Regenerative Medicine,Centro de Investigación Príncipe Felipe,Valencia 46012,Spaina b s t r a c ta r t i c l e i n f o Article history:Received 4March 2011Revised 29March 2011Accepted 11April 2011Available online 16April 2011Keywords:Nitric oxide MMPEMMPRINIschemia/reperfusion MiceFree radicalsNitric oxide (NO)is an important defense against myocardial ischemia/reperfusion (I/R)injury.Although matrix metalloproteinase (MMP)-mediated necrosis of cardiac myocytes is well characterized,the role of inducible NO synthase (iNOS)-derived NO in this process is poorly understood.I/R injury was increased in iNOS-de ficient mice and in mice treated with 1400W (a pharmacological iNOS inhibitor)and was associated with signi ficantly increased expression of extracellular matrix metalloproteinase inducer (EMMPRIN)and EMMPRIN-associated MMPs.Transcriptional activity of an EMMPRIN luciferase promoter reporter expressed in cardiac myocytes was inhibited by NO in a cGMP-dependent manner,and this transcriptional inhibition was abolished by mutation of a putative E2F site.Consistent with these findings,EMMPRIN null mice,in which iNOS is normally induced,are partially protected against I/R injury.Pharmacological inhibition of iNOS in EMMPRIN null mice had no additional protective effect,suggesting that EMMPRIN is a downstream target of NO.Administration of anti-EMMPRIN neutralizing antibodies partly reduced the excess heart damage and MMP-9expression induced by I/R in iNOS null mice,indicating that regulation of EMMPRIN is an important mechanism of NO-mediated cardioprotection.©2011Elsevier Inc.All rights reserved.Inducible nitric oxide (NO)synthase (iNOS)plays a pivotal role in cardioprotection [1,2].NO-mediated cardioprotection in response to ischemia/reperfusion (I/R)injury is implicated in several molecular pathways,including NF-κB transactivation of COX2[3,4],PKC-δphosphorylation [5],nitration of PKC-ε[6],and mitochondrial perme-ability transition [7].However,the role of NO in cardiac cell necrosis associated with extracellular matrix degradation,an early step in I/R injury,is poorly understood.Extracellular matrix metalloproteinases (MMPs)are proteolytic degrading enzymes that cleave extracellular matrix components,chemokines,vasoactive factors,cytokines,growth factors,angiogenic factors,and adhesion molecules [8].In the heart,activation of MMPs triggers cardiac myocyte necrosis and left-ventricle dysfunction and promotes adverse left-ventricle remodeling [9].Consequently,strat-egies aimed at inhibiting MMP proteolytic activation may provide an effective means of avoiding cardiac injury.Recently,it has been shown that EMMPRIN (extracellular matrix metalloproteinase inducer;also known as CD147and basigin)regulates the expression and activation of several MMPs,including MMP-2,MMP-9,and MT1-MMP,and is expressed in monocytes,cardiac myocytes,andendothelial cells [10,11].Interventions targeting EMMPRIN may there-fore inhibit not just one endopeptidase but a broad spectrum of MMPs implicated in I/R injury.To investigate the possible effects of NO-mediated repression of EMMPRIN,we induced I/R injury in mice subjected to coronary artery occlusion followed by 24-h reperfusion.We found that NO represses EMMPRIN transcription,providing signi ficant evidence about the possible role of EMMPRIN in the recovery of cardiac function.Materials and methods ReagentsGeneral cell culture supplies were from BD Biosciences (Spain),calf serum was from BioWhittaker (Verviers,Belgium),and cell culture and antibiotics were from Sigma (St.Louis,MO,USA).Conjugated secondary antibodies were from GE HealthCare (Spain).EDTA-free protease inhibitor cocktail tablets were from Roche (Spain).OptiMem and Lipofectamine were from GIBCO BRL.(Z )-1-[2-(2-Aminoethyl)-N -(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA-NO),S -nitroso-N -acetyl-D,L -pen-icillamine,(Z )-1-{N -[3-aminopropyl]-N -[4-(3-aminopropylammonio)butyl]-amino}-diazen-1-ium-1,2-diolate,and N -[3-(aminomethyl)ben-zyl]acetamidine (1400W)were from Alexis Biochemicals (San Diego,CA,USA).Rp-8-Br-PET-cGMPS was from Biolog (Germany).Anti-MMP-9and anti-MMP-2were from BD Biosciences;anti-EMMPRIN and anti-Free Radical Biology &Medicine 51(2011)387–395⁎Corresponding author at:Dept.of Atherothrombosis and Cardiovascular IC.Sinesio Delgado 3,Madrid 28029,Spain.Fax:+34914531245.E-mail address:czaragoza@cnic.es (C.Zaragoza).1These authors contributed equally to thisarticle.0891-5849/$–see front matter ©2011Elsevier Inc.All rights reserved.doi:10.1016/j.freeradbiomed.2011.04.021Contents lists available at ScienceDirectFree Radical Biology &Medicinej o u r n a l h o me p a g e :w w w.e l s e v i e r.c om /l oc a t e /fr e e r a d b i o me dEMMPRIN control IgG were from Serotec.Anti-MAC3antibody was from BD Pharmingen(Spain).MiceiNOS null mice(B6;129P2-Nos2tm1Lau/J)and the corresponding wild-type controls were purchased from The Jackson Laboratory(Bar Harbor,ME,USA).No differences in size and weight were detected between these mice.All animals were housed in our animal facilities in isolated rooms.The local ethics review board approved animal procedures.EMMPRIN null mice were kindly donated by Dr.Alicia García Arroyo.The study conformed to the Guidelines for the Care and Use of Laboratory Animals published by the U.S.National Institutes of Health(NIH Publication No.85-23,revised1996).CellsThe HL1cardiomyocyte cell line was kindly donated by Dr.Antonio Bernad and cultured as described[12].The murine macrophage cell line RAW247.6was grown as described[13].Ischemia/reperfusion of the coronary arteryIschemia was induced by coronary artery ligation.Twelve-week-old mice were intraperitoneally anesthetized with ketamine/xylazine (100and10mg/kg,respectively),intubated with a1-mm steel tube, and ventilated(2ml,80strokes/min).The thorax was opened between the second and the third ribs and widened with the aid of a mouse retractor.The pericardium was opened and the main left coronary artery was occluded for30min close to its bifurcation by using a6-0silk suture.Decoloration of the ventricle provided direct evidence of coronary occlusion.Ligation was then released,the chest closed,negative pressure restored,and the skin sutured.Control mice (sham)were included in the assays,in which the same procedure was performed excepting the occlusion of the left coronary artery.For in vivo iNOS inhibition,iNOS wild-type mice were injected in the tail vein with2mg/kg/day1400W,30min before ischemia and 24h after ischemia/reperfusion.To neutralize EMMPRIN in vivo,mice were administered a range of doses(100–500mmol/L/kg)of anti-EMMPRIN antibodies or control IgG's by intravenous tail injection.A dose of250μmol/L/kg of the anti-EMMPRIN antibody was found to reduce MMP-9expression by more than50%compared with control IgG and was used in subsequent experiments.To evaluate the effects of anti-EMMPRIN antibody on ischemia/reperfusion,antibodies were administered4days before surgery and reperfusion,after which infarct size,heart function,and the expression of EMMPRIN and MMP-9were evaluated.Nitrite assayNitrite concentrations were determined by a modification of the Griess assay[14].Briefly,50-μl samples and sodium nitrite standards were incubated with an equal volume of Griess reagent(1% sulfanilamide,0.1%naphtylethylenediamine,and 2.5%H3PO4)for 10min at room temperature.The absorbance of samples at540nm was measured in a microplate reader.Fluorescence molecular tomographyFluorescence molecular tomography was performed with a chamber imaging system(FMT1500;VisEn Medical,PerkinElmer,USA),which delivers true3D spatial information aboutfluorochrome distribution and concentration.Spatial encoding is achieved by varying the spatial location of the excitation laser,wherebyfluorochromes are excited by 80different point sources distributed over the area of interest behind the imaging chamber,and the resultingfluorescence is recorded by a CCD camera.Light is directed at various projections,and signal is detected from multiple points of the sample surface.A postprocessing algorithm then uses light propagation properties and the information in these80raw images to tomographically reconstruct the location and concentration of thefluorochromes.To detect areas of inflammation,we injected the imaging probe Prosense-680(VisEn;5nmol in150μl PBS; excitation wavelength680±10nm,emission700±10nm),which detects cathepsin-B activity,into the tail vein24h before imaging. Tomographic images were acquired in anesthetized mice,and the amount of cathepsin-B activity was evaluated with the aid of the FMT 1500software package.EchocardiographyMouse hearts were visualized by echocardiography over time,using a high-frequency micro-ultrasound system(Vevo770;VisualSonics, Toronto,ON,Canada).During all experiments,mice were anesthetized with1.5%isoflurane gas,which resulted in a heart rate of approximately 300beats/min.Anesthetized animals were placed on the Integrated Rail System and Mouse Handling Table(provided by the manufacturer), which allows simultaneous acquisition of temperature data(37°C,by using the integrated heating pad)and electrocardiography(with a lead II configuration)throughout the experiment.The chest of the mouse was carefully shaved,and warm ultrasound transmission gel was applied to ensure optimal image quality.Parasternal short-axis-view images of the heart were recorded using a30-MHz RMV scan head in a B-mode to allow M-mode recordings by positioning the cursor in the parasternal short-axis view perpendicular to the interventricular septum and posterior wall of the left ventricle.From these recordings, the following parameters were determined using the on-site software cardiac package(VisualSonics):left-ventricle end-diastolic diameter, left-ventricle end-diastolic volume,ejection fraction,and shortening fraction.Histology and immunohistochemistryMouse hearts were embedded in paraffin and4-μm serial sections were obtained as described[13].Heart morphology was visualized by eosin–hematoxylin staining,and collagen deposition was detected by Masson's trichrome staining.Immunohistochemical detection of EMMPRIN,MMP-9,and MMP-2was performed by incubating samples with the corresponding primary and secondary antibodies and detecting boundfluorescence-conjugated secondary antibodies by confocal microscopy as described [15].Double immunostaining of EMMPRIN and macrophages was performed in heart sections by confocal microscopy with anti-EMMPRIN and anti-MAC3antibodies.In brief,heart samples were embedded in paraffin,and4-mm-thick serial sections were subjected to confocal microscopy with the appropriate antibodies[13].Gelatin zymographyProtein lysates prepared from hearts were resolved on8.5–10%SDS–polyacrylamide gels containing1mg/ml gelatin.Gels were incubated for 30min with renaturing buffer(2.5%Triton X-100)and then incubated overnight with developing buffer(50mM Tris–HCl,pH7.5,200μM NaCl,10μM CaCl2,0.02%Brj35).Gels were stained with Coomassie blue for1h,and gelatinolytic activity was detected after incubation with destaining solution(methanol:acetic acid:water,50:10:4).Cloning of murine EMMPRIN regulatory regionsA1-kb DNA fragment upstream of the mouse EMMPRIN coding sequence was cloned by PCR using the commercial BAC CH29-603O5 kit(which contains part of mouse chromosome10)from CHORI388 C.Tarin et al./Free Radical Biology&Medicine51(2011)387–395(Children's Hospital Oakland Research Institute)and the following primers:forward,5′-CGGGGTACCAGCACTCCATCCAAAGGCAGA-3′; reverse,5′-GGAAGATCTGTCGCCTCGTCCAGGAGC-3′.The resulting fragment was cloned into the pGL3-Basic vector(Promega),upstream of a luciferase reporter gene(pEMMPRIN-WT).Mutagenesis of the EMMPRIN promoterpEMMPRIN-WT(p1000)was used as the template in PCR assays to create the following serial deletions of the EMMPRIN promoter:p500, a plasmid containing the proximal500bp of the EMMPRIN promoter sequence;p250,containing the proximal250bp of the sequence; p1000-500,containing the distal500bp;p1000-750,containing the distal250bp;and p875-750,containing the sequence corresponding to positions875–750of the EMMPRIN promoter.In addition,point mutations(substitutions)with affinity for the E2F transcription factor were introduced into the binding site,using the Stratagene site-directed mutagenesis kit.The following combination of primers was used (mutated bases in lowercase):forward,5′-GGGGTTAGAAGCCTtCtCtA-CAGTGCACGACCTTCAAA-3′;reverse,5′-TTTGAAGGTCGTGCACTGTaGa-GaAGGCTTCTAACCCC-3′.Transient transfectionCells were transfected and luciferase content was measured as previously described[16].Briefly,cells were transiently transfected with1μg of DNA and10μl of Lipofectamine2000for4h,washed,and incubated with MEM/10%FCS for16h.Cells were cotransfected with a plasmid containing the constitutive promoter region of CMV fused to the Renilla luciferase gene as a control.All samples were analyzed for firefly(Photinus pyralis)and renilla(Renilla reniformis)luciferase content.Results were normalized to Renilla luciferase content.Statistical analysisUnless otherwise specified,data are expressed as means±SD.Cell culture experiments were performed in triplicate,and conditions were assayed in duplicate on each replicate.Animal experiments were performed in triplicate,and the numbers of animals and replicates are specified in the text.Whenever comparisons were made with a common control,significance of differences was tested by analysis of variance followed by Dunnett's modification of the t test.Differences were considered significant at p b0.05.Error bars represent±SD.ResultsIschemia/reperfusion injury is more severe in mice lacking iNOSWild-type and iNOS-deficient mice were subjected to I/R injury by left coronary artery occlusion(see Materials and methods).Cardiac expression and activity of iNOS were evaluated,revealing a significant increase in iNOS expression in wild-type hearts in response to injury (Fig.1A).To detect areas of inflammation,mice were injected24h before injury with5nmol of the pancathepsin protease sensor Prosense-680and examined byfluorescence molecular tomography. The area of inflammation was significantly smaller in hearts of wild-type mice compared with iNOS-deficient mice(Fig.1B).Similarly, high-frequency ultrasound revealed smaller infarct sizes and better ejection fraction values in wild-type hearts(Figs.1C and D).Table1 lists the cardiac parameters collected after M-mode ultrasound evaluation of mice.Expression of EMMPRIN is increased in iNOS null mice after I/R injuryMatrix metalloproteinases are implicated in cardiac necrosis in response to I/R,by inducing extracellular matrix(ECM)degradation.Among the signals that prompt expression in the heart,it was recently described that the extracellular matrix metalloproteinase inducer EMMPRIN is increased in human left ventricle after acute myocardial infarction[17]and regulates the secretion of MMPs by cardiac cells[10]. We found that in the hearts of iNOS null mice,I/R induced the expression and secretion of MMP-2and MMP-9,together with a significantincrease ck of NO increases myocardial damage during ischemia/reperfusion.Ischemia (30min)was induced by left coronary artery occlusion in the hearts of wild-type and iNOS null mice(n=6mice/group/triplicate)and was followed by reperfusion for24h.(A)Immunoblot showing iNOS protein expression,with GAPDH as loading control,and NO production(right).Data are means±SD;*p b0.05,iNOS WT+I/R vs iNOS WT−I/R.(B)Cathepsin-B activity in operated hearts detected byfluorescence molecular tomography(mean±SD;*p b0.05,iNOS WT I/R vs iNOS KO I/R).(C)Infarct size (mean±SD;*p b0.05,iNOS WT vs iNOS KO).(D)Ejection fraction(mean±SD; *p b0.05,iNOS WT I/R vs iNOS KO I/R).Table1Ultrasound parameters.WT control WT I/R iNOS control iNOS I/RLVDS(mm) 3.07±0.3 3.50±0.6 3.06±0.28 4.10±0.12 LVDD(mm) 4.17±0.40 4.66±0.11 4.24±0.29 5.56±0.49VS(ml)37.24±3.0753.00±6.9134.43±6.9459.78±3.82VD(ml)57.43±2.0868.63±4.8159.45±10.4288.60±15.28* SV(ml)20.19±4.0115.63±6.4725.02±3.6228.82±6.39* EF(%)51.26±3.7042.49±4.6251.9±4.1530.53±3.35* FS(%)25.55±2.2120.66±4.8126.30±1.9814.94±2.51* Heart rate(bpm)326±11.77339±23.33355±8.98343±60.80 Weight(mg)120±6.59132±6.12110±9.09165±19.25 LVDS,left-ventricle end-systolic diameter;LVDD,left-ventricle end-diastolic diameter; VS,systolic volume;VD,diastolic volume;SV,systole–diastole volume difference;EF, left-ventricle ejection fraction;FS,shortening fraction.*p b0.05,WT I/R vs iNOS I/R.389C.Tarin et al./Free Radical Biology&Medicine51(2011)387–395in EMMPRIN,compared to wild-type mice(Figs.2A and B,respectively), suggesting that NO may regulate the expression and secretion of MMPs, at least,by controlling the levels of EMMPRIN in hearts subjected to I/R.EMMPRIN-induced MMP activation is related to the glycosylation of the protein.High-glycosylated(HG)forms of EMMPRIN(40–60kDa), rather than low-glycosylated(LG)forms(25–30kDa),induce MMP-mediated ECM degradation[18].In mouse heart lysates,three main forms of EMMPRIN were detected,and exogenous administration of NO to iNOS null mice(tail vein injection of20μg/kg/day sodium nitroprusside(SNP))significantly reduced the levels of both HG and LG forms of EMMPRIN in response to I/R(Fig.2C),suggesting that NO may regulate EMMPRIN in the heart at the transcriptional level, although direct NO-induced EMMPRIN posttranscriptional modifica-tion cannot be excluded.NO inhibits the expression of EMMPRIN mRNA in mouse heart during I/R To explore the contribution of NO to EMMPRIN transcription in response to I/R,we measured EMMPRIN mRNA by RT-PCR in total RNA isolated after surgery from wild-type and iNOS null mouse hearts.I/R induced a significantly higher increase in EMMPRIN mRNA in iNOS null mice compared with their wild-type counterparts(Fig.3A),suggesting that iNOS-derived NO might repress the transcription of EMMPRIN in the heart.To investigate the specificity of this NO action in cardiomyocytes, we compared EMMPRIN mRNA expression in the cardiac myocyte cell line HL-1B and in RAW247.6macrophages treated with NO donors. Administration of DETA-NO significantly reduced the levels of EMM-PRIN mRNA in HL-1B cells(Fig.3B),whereas in macrophagesneitherFig.2.NO inhibits the expression of EMMPRIN and MMP-9during ischemia/reperfusion.(A)Left:immunoblot detection of MMP-9,MMP-2,EMMPRIN,and GAPDH in heart lysates from wild-type and iNOS null mice subjected to IR.Right:densitometric analysis(relative to GAPDH band intensities)from three independent immunoblots(n=6mice/group/ triplicate,mean±SD;*p b0.05,MMP-9WT IR vs MMP-9KO IR;#p b0.05,EMMPRIN WT IR vs EMMPRIN KO IR).(B)Gelatinolytic activity of MMP-9and MMP-2.Right:densitometric analysis of enzymatic activities detected in three independent assays(n=6mice/group/triplicate,mean±SD;*p b0.05,MMP-9WT IR vs MMP-9KO IR;#p b0.05,MMP-2WT IR vs MMP-2KO IR).(C)Expression of EMMPRIN in iNOS null mice subjected to sham operation(C),ischemia/reperfusion(I/R),and I/R initiated after treatment with the NO donor sodium nitroprusside(20μg/kg/day)(I/R NO);SNP treatment was continued for24h after I/R.Right:densitometric analysis(relative to GAPDH band intensities)from three independent immunoblots of the levels of each form of EMMPRIN(40,37,23kDa;Total,total intensity/lane;n=6mice/group/triplicate;mean±SD;*p b0.05,40kDa IR vs40kDa IR NO;#p b0.05,37kDa IR vs37kDa IR NO;¢p b0.05,23kDa IR vs23kDa IR NO;@p b0.05,Total IR vs Total IR NO.390 C.Tarin et al./Free Radical Biology&Medicine51(2011)387–395DETA-NO nor lipopolysaccharide (LPS)-induced expression of iNOS affected the expression of EMMPRIN mRNA (Fig.3C).Confocal immuno-fluorescence assay on mouse heart sections con firmed that EMMPRIN expression in in filtrating immune cells did not differ between I/R-injured wild-type and iNOS null mice (Fig.3D),suggesting that NO-mediated transcriptional repression of EMMPRIN occurs speci fically in cardiac myocytes.NO induces transcriptional repression of the EMMPRIN promoter in cardiac myocytesTo analyze NO-induced transcriptional repression of EMMPRIN in cardiac myocytes,we fused the 1-kb sequence upstream from the EMMPRIN coding sequence to the reporter fire fly luciferase in the pGL3-Basic vector (pEMMPRIN-WT).In transiently transfected cardiac myocytes,DETA-NO signi ficantly reduced EMMPRIN promoter activity in a dose-dependent manner (Fig.4A),whereas in transfected macrophages NO (either endogenously produced by stimulation with LPS or exogenously administered as DETA-NO)had no signi ficant effect (Fig.4B).The soluble cGMP analog 8-Br-cGMP mimicked the effect of DETA-NO on cardiac myocytes,suggesting that NO represses EMMPRIN transcription via the cGMP/cGMP-dependent protein kinase (PKG)pathway (Fig.4C).Consistent with this idea,pharmacological inhibition of PKG with Rp-8-Br-PET-cGMPS (PET)stimulated EMMPRIN promoter activity in control cells (Fig.4D)and partially restored promoter activityin DETA-NO-treated cells (Fig.4D).Furthermore,overexpression of a dominant positive form of PKG-1α(fG1AC)[16]signi ficantly reduced EMMPRIN promoter activity in cardiac myocytes expressing pEMM-PRIN-WT (Fig.4D).These results indicate that NO limits the transcription of EMMPRIN via the cGMP/PKG pathway in cardiac myocytes,and this molecular pathway might contribute to the cardioprotective effect of NO.To examine the mechanism of NO-induced transcriptional inhibi-tion of EMMPRIN,we tested the effects of NO on a series of deletion mutants of the promoter (Fig.5).Experiments in transfected cardiac myocytes showed that two of the vectors,p1000-750(containing the distal 250bp of the EMMPRIN promoter)and p875-750(containing the 125-bp region between bp −875and bp −750of the EMMPRIN promoter),contain the region responsible for NO-induced transcrip-tional repression (Figs.5A,bottom,and B).Detailed analysis of this region revealed binding sites for several transcription factors,including one for E2F,which has been reported to mediate transcriptional repression of several genes [19].Mutation of the E2F siteabolishedFig.3.I/R increases EMMPRIN mRNA expression in iNOS-de ficient mouse hearts.(A)Real-time quantitative RT-PCR showing expression of EMMPRIN in the hearts of wild-type and iNOS null mice after I/R (n =3/triplicate,mean ±SD;*p b 0.05,iNOS null +I/R vs iNOS null −I/R).(B)Real-time quantitative RT-PCR showing the expression of EMMPRIN in cultured HL1cardiac myocytes in response to treatment with 100μM DETA-NO (n =3,mean ±SD;*p b 0.05).(C)Real-time quantitative RT-PCR showing the expression of EMMPRIN in cultured RAW 247.6macrophages in response to exogenous treatment (16h)with 100μM DETA-NO,endogenous production of NO (50μM LPS),or inhibition of endogenous iNOS with 100μM 1400W (n =3mice,mean ±SD).(D)Confocal microscopy detection of immune cells (anti-Mac-3,green)expressing EMMPRIN (red),in heart in filtrates after I/R.Nuclei were stained with DAPI (blue)(n =3mice/triplicate).Scale bars,50μm.Fig.4.NO represses EMMPRIN transcription in cardiac myocytes via the NO/cGMP/PKG pathway.(A)Dose response to the NO donor DETA-NO of EMMPRIN transcriptional activity in cardiomyocytes transiently transfected with a luciferase reporter construct of the 1000-bp EMMPRIN promoter region (pEMMPRIN-WT;n =10,mean ±SD;*p b 0.05vs 0).(B)Effects of exogenous NO (100μM DETA-NO),endogenous production of NO (50μM LPS),and pharmacological inhibition of LPS-induced iNOS (100μM 1400W)on EMMPRIN transcriptional activity in RAW 247.6macrophages transiently transfected with pEMMPRIN WT (n =3,mean ±SD).(C)Dose response to the cGMP analog 8-Br-cGMP in cardiac myocytes transiently transfected with pEMMPRIN-WT (n =10,mean ±SD;*p b 0.05vs 0).(D)Effect of PKG on EMMPRIN transcriptional activity in cardiac myocytes expressing pEMMPRIN-WT and either cotransfected with dominant-positive PKG-1α(fG1AC)or treated with 10μM PKG inhibitor PET in the presence or absence of 100μM DETA-NO (n =3,mean±SD;*p b 0.05,vs transfected resting cells (−).391C.Tarin et al./Free Radical Biology &Medicine 51(2011)387–395NO-induced repression of EMMPRIN promoter activity,suggesting that this factor is a target of NO (Fig.5B).Targeted disruption of EMMPRIN promotes cardiac protection of mouse heartsTo investigate the role of EMMPRIN in NO-mediated cardioprotec-tion in vivo,we induced I/R in EMMPRIN-de ficient mice.In response to injury,induction of iNOS (Fig.6A)and infarct size were similar to those found in wild-type animals (Fig.6B).Pharmacological inhibition of iNOS with 1400W increased the expression of EMMPRIN (Fig.6C,WT I/R vs WT 1400W I/R)and MMP-9(Fig.6D,WT I/R vs WT 1400W I/R)in wild-type hearts over the levels found in mice lacking EMMPRIN (Figs.6C and D,KO I/R vs KO I/R 1400W I/R),and interestingly,1400W had no signi ficant effect on heart contractility in the absence of EMMPRIN(Fig.6E,KO I/R vs KO 1400W I/R).These results suggest that EMMPRIN is a target in the cardioprotective action of NO (Fig.6F).Inhibition of EMMPRIN in iNOS null mice improves heart contractility after I/RTo explore the potential protection of targeted inhibition of EMMPRIN against I/R injury,we administered speci fic anti-EMMPRIN antibodies (which speci fically bind to an epitope located at the N-terminal Ig extracellular domain)to iNOS null mice subjected to I/R.Anti-EMMPRIN antibodies signi ficantly downregulated expression of MMP-9compared with control mice or mice injected with IgG1(Fig.7A).Treatment with anti-EMMPRIN antibodies did not alter EMMPRIN expression (Fig.7B);but the performance of the heart was signi ficantly recovered compared to untreated controls (Fig.7C,left).Anti-EMMPRIN antibodies were also bene ficial for cardiac contractility in wild-type mice (Fig.7C,right).Fig.5.Transcriptional repression of the EMMPRIN promoter by NO is mediated at least in part through E2F.(A)Effects of NO on EMMPRIN promoter activity in cardiomyocytes transiently transfected with pEMMPRIN-WT (p1000)or a series of deletion mutants.Top:transcriptional activity of p1000and p500(n =3,mean±SD;*p b 0.05,vs p1000).Middle:transcriptional activity of p1000and p250(n =3,mean ±SD;*p b 0.05,p1000+DETA-NO vs p1000;#p b 0.05,vs p1000).Bottom:transcriptional activity of p1000-500and p1000-750,which contain the distal 500and 250bp of p1000,respectively (n =3,mean±SD;*p b 0.05,vs p1000-500;#p b 0.05,vs p1000-750).(B)Transcriptional activity of p875-750and a point mutation in the putative E2F binding site (pE2F;see Materials and methods;n =3,mean±SD;*p b 0.05,p875-750vs p875-750+NO).392 C.Tarin et al./Free Radical Biology &Medicine 51(2011)387–395These findings indicate that NO-mediated cardioprotection is at least regulated by inhibition of EMMPRIN in mouse hearts,pointing toward EMMPRIN as a candidate for the treatment of myocardial infarction.DiscussionOur results show that iNOS is expressed in response to I/R in mice,and the resulting production of NO induces cGMP/PKG-mediated transcriptional repression of EMMPRIN,restricting the activation of extracellular MMPs.NO-mediated transcriptional repression is restrict-ed to cardiac myocytes of hearts subjected to I/R and is dependent on the integrity of a binding site for the transcription factor E2F in the EMMPRIN promoter.The hearts of EMMPRIN null mice express iNOS normally in response to I/R,but show signi ficantly less heart damage and lower expression of MMP-9than wild-type hearts.Inhibition of iNOS in EMMPRIN null mice did not protect cardiac contractility or reduce MMP-9expression,pointing to EMMPRIN as a target of iNOS in cardiac protection.Furthermore,protection of cardiac function was prompted with anti-EMMPRIN neutralizing antibodies before I/R.NO is an important defense against I/R injury [20].Several mechanisms by which NO may exert cardiac protection have been identi fied through the use of NO donors or adenoviral delivery of iNOS [4].However,little is known about the mechanisms elicited by endogenous NO in response to I/R or the role of NO-mediated extracellular matrix degradation in this context.Our results indicate that NO-mediated EMMPRIN downregulation in cardiac myocytes is an important mecha-nism in cardiac protection,preserving heart contractility by limiting ECM degradation.NO derived from iNOS or endothelial NOS plays a role in cardiac protection [2,21,22]and regulates the expression of several genes by mechanisms that include the cGMP/PKG pathway [16,23].Our results show that NO-mediated transcriptional repression of EMMPRINinvolvesFig.6.EMMPRIN-de ficient mice show reduced heart damage in response to I/R.(A)Immunoblot detection of iNOS (and GAPDH as control)in wild-type (WT)and EMMPRIN null mice (KO)subjected to I/R (n =3mice/group/triplicate).(B)I/R-induced heart damage in wild-type and EMMPRIN null mice (n =5mice/group/duplicate;mean ±SD;*p b 0.05).(C)Immunohistochemical detection of EMMPRIN in WT and EMMPRIN KO mice subjected to I/R.Where indicated,animals were treated with 100μM 1400W (see Materials and methods;n =3mice/group/triplicate;scale bars,50μm).(D)Immunohistochemical detection of MMP-9in the same sections as in (C).(E)Left-ventricle ejection fraction values obtained from control and post-I/R wild-type and EMMPRIN null mice,treated or not with 100μM 1400W (mean ±SD;*p b 0.05,WT C vs WT IR;#p b 0.05,WT IR vs WT IR 1400W;¢p b 0.05,KO C vs KO IR 1400W;@p b 0.05,WT IR 1400W vs KO IR 1400W).(F)Proposed mechanism of NO-mediated EMMPRIN inhibition in cardiac protection.393C.Tarin et al./Free Radical Biology &Medicine 51(2011)387–395。