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Effects of recombinant retroviral vector mediated human insulin like growth factor-1 gene transfection on skeletal muscle growth in rat

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Chinese Medical Journal 2006年第23期119卷 1991-1998页

作者： RONG Shu-Ling LU Yong-Xin LIAO Yu-hua WANG Xiao-Lin GUO He-Ping CHANG Chao GAO Yan-Zhang mi shao-hua Wan Jian-Ping Department of Cardiology Union Hospital Tongji Medical CollegeHuazhong University of Science and Technology Wuhan 430022China Department of Paediatrics Heping Hospital Changzhi Medical College Changzhi 046000 China

Background This study transferred a recombinant gene encoding human insulin like growth factor-1 （hIGF-1） into modified primary skeletal myoblasts with a retroviral vector （pLgXSN） and determined whether the hIGF-1 promoted growth of skeletal muscle in rat.Methods hlGF-lcDNA was amplified in vitro from normal human liver cells by using RT-PCR and cloned into plasmid vector pLgXSN. The recombinant vector pLghIGF-1SN and control vector pLgGFPSN were transfected into packaging cell PT67 and G418 was used to select positive colony. Myoblasts were infected with a high titre viral supernatant and transduction efficiency was evaluated as GFP expression. The expression of hIGF-1 mRNA in myoblasts was investigated by immunocytochernistry and RT-PCR. MTT assays detected the growth of myoblasts in vitro. Myoblasts transduced with pLghlGF-1SN were injected into hind limb muscles of 10-12 week male SD rats. Formed tissues were harvested 4 weeks later. Myocyte diameter, mean weight of hind limb and body were measured to evaluate the skeletal muscle growth. Results Recombinant retroviral plasmid vector pLghlGF-1SN was constructed successfully. The titre of the packaged recombinant retrovirus was 1 × 106 cfu/ml. The transfection rate of PT67 cells reached 100% after G418 screening, hIGF-1 expression was positive in myoblast-IGF-1. The proliferation rate of myoblast-IGF-1 in vitro was higher than GFP-myoblast or myoblast （P〈 0.05）. The mean weights of hind limb and body of rats injected myoblast-IGF-1 were higher than those of the rats injected with myoblast-GFP or myoblast （P〈 0.05）. Myocyte diameter had a significant increase in IGF-1 group compared to GFP group and myobiast group （P〈 0.05）. Conclusions The transfection of the human IGF- 1 gene mediated by a retroviral vector can promote the growth of skeletal muscle in rats. Genetically modified primary skeletal myoblasts provide a possibly effective approach to treat some skeletal muscle diseases.

关键词： insulin like growth factor-1 skeletal muscle growth retroviral vector

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Effects of transplanted myoblasts transfected with human growth hormone gene on improvement of ventricular function of rats

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Chinese Medical Journal 2008年第4期121卷 347-354页

作者： RONG Shu-ling LU Yong-xin LIAO Yu-hua WANG Xiao-lin WANG Yong-jin CHANG Chao WANG Yu-qin LIU Qi-yun GAO Yan-zhang mi shao-hua Department of Cardiology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei 430022 China Department of Cardiology Heping Hospital Changzhi Medical College Changzhi Shanxi 046000 China

Background Cell transplantation for myocardial repair is limited by early cell death. Gene therapy with human growth hormone （hGH） has been shown to promote angiogensis and attenuate apoptosis in the experimental animal. This study was conducted to explore the effects of myoblast-based hGH gene therapy on heart function restoration and angiogenesis after myocardial infarction, and to compare the differences between myoblast-based hGH gene therapy and myoblast therapy.Methods Myoblasts were isolated from several SD rats, cultured, purified, and transfected with plasmid pLghGHSN and pLgGFPSN. Radioimmunoassay （RIA） was used to detect the expression of hGH in these myoblasts. SD rats underwent the ligation of the left anterior descending coronary artery so as to establish a heart ischemia model. Thirty surviving rats that underwent ligation were randomly divided into 3 equal groups 2 weeks after left coronary artery occlusion： pLghGHSN group received myoblast infected with hGH gene transplantation; pLgGFPSN group received myoblast infected with GFP gene transplantation; control group： received cultured medium only. Four weeks after the injection the surviving rat underwent evaluation of cardiac function by echocardiography. The rats were killed and ventricular samples were undergone immunohistochemistry with hematoxylin-eosin and factor Ⅷ. Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to examine the mRNA expression of vascular endothelial growth factor （VEGF）, bax and Bcl-2. hGH expression in myocardium was examined by Western blot.Results Myoblast can be successfully isolated, cultured and transfected. The expression of hGH in transfected myoblast was demonstrated with RIA. Four weeks after therapy, the cardiac function was improved significantly in pLghGHSN group and pLgGFPSN group. Fractional shortening （FS） and ejection fraction （EF） i

关键词： heart failure myoblast growth hormone gene therapy

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Recombinant proteins secreted from tissue-engineered bioartificial muscle improve cardiac dysfunction and suppress cardiomyocyte apoptosis in rats with heart failure

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Chinese Medical Journal 2010年第24期123卷 3626-3633页

作者： RONG Shu-ling WANG Yong-jin WANG Xiao-lin LU Yong-xin WU Yin LIU Qi-yun mi shao-hua XU Yu-lan Department of Cardiology Heping Hospital Changzhi Medical College Changzhi Shanxi 046000 China Department of Paediatrics Heping Hospital Changzhi Medical College Changzhi Shanxi 046000 China Department of Cardiology Union Hospital Tongji Medical College Huazhong University of Science and Tecnology Wuhan Hubei 430022 China Department of Psychology School of Educational Science South China Normal University Guangzhou Guangdong 510631 China

Background Tissue-engineered bioartificial muscle-based gene therapy represents a promising approach for the treatment of heart diseases. Experimental and clinical studies suggest that systemic administration of insulin-like growth factor-1 （IGF-1） protein or overexpression of IGF-1 in the heart exerts a favorable effect on cardiovascular function. This study aimed to investigate a chronic stage after myocardial infarction （mi） and the potential therapeutic effects of delivering a human IGF-1 gene by tissue-engineered bioartificial muscles （BAMs） following coronary artery ligation in Sprague-Dawley *** Ligation of the left coronary artery or sham operation was performed. Primary skeletal myoblasts were retrovirally transduced to synthesize and secrete recombinant human insulin-like growth factor-1 （rhIGF-1）, and green fluorescent protein （GFP）, and tissue-engineered into implantable BAMs. The rats that underwent ligation were randomly assigned to 2 groups： mi-IGF group （n=6） and mi-GFP group （n=6）. The mi-IGF group received rhIGF-secreting BAM （IGF-BAMs） transplantation, and the mi-GFP group received GFP-secreting BAM （GFP-BAMs） transplantation. Another group of rats served as the sham operation group, which was also randomly assigned to 2 subgroups： S-IGF group （n=6）and S-GFP group （n=6）. The S-IGF group underwent IGF-1-BAM transplantation, and S-GFP group underwent GFP-BAM transplantation. IGF-1-BAMs and GFP-BAMs were implanted subcutaneously into syngeneic rats after two weeks of operation was performed. Four weeks after the treatment, hemodynamics was performed. IGF-1 was measured by radioimmunoassay, and then the rats were sacrificed and ventricular samples were subjected to immunohistochemistry. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to examine the mRNA expression of bax and Bcl-2. TNF-α and caspase 3 expression in myocardium was examined by Western *** Primary rat myoblasts were retrovirally transduced to secre

关键词： heart failure muscles insulin-like growth factor-1 gene therapy

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船载雷达海冰监测及预报技术在辽东湾冰区油轮外输作业中的应用

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海洋环境科学 2006年第1期25卷 76-79页

作者：肖井坤邵秘华张勐大连海事大学环境科学与工程学院辽宁大连116026

渤海的辽东湾是世界上典型的海湾结冰区,位于冰区中的石油平台油轮外输作业中,流冰连续不断地漂移、聚集形成堆积冰,给海上油轮的外输作业安全带来严重危害。本文概述了在国内首次利用船载雷达,在该湾冰区石油平台上,现场采用雷达海冰... 详细信息

渤海的辽东湾是世界上典型的海湾结冰区,位于冰区中的石油平台油轮外输作业中,流冰连续不断地漂移、聚集形成堆积冰,给海上油轮的外输作业安全带来严重危害。本文概述了在国内首次利用船载雷达,在该湾冰区石油平台上,现场采用雷达海冰监测预报技术系统,进行遥感海冰数据源的获取,图像的解译和数值化处理,得到流冰类型、冰厚度、流冰密集度、流冰速度、方向、流冰漂移轨迹等要素。同时首次现场实时计算出雷达冰漂流场矢量图,并开展了雷达海冰数值跟踪预报,在冰区油轮外输作业中得到很好应用,为雷达技术对水体和海冰监测和预报、遥感应用新领域提供科学依据。

关键词：船载雷达海冰预报冰漂流遥感系统辽东湾

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现场分析海水中氨氮的靛酚蓝法

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大连海事大学学报 2007年第1期33卷 142-145页

作者：陈燕邵秘华林小燕蒋岳文大连海事大学环境科学与工程学院辽宁大连116026 国家海洋环境监测中心辽宁大连116023

采用正交实验对显色试剂配比进行优化,并探讨了水质分析理化条件,确定了现场分析氨氮的靛酚蓝法.其结果表明:复合试剂中柠檬酸钠含量为120 mg/ml,EDTA含量为30 mg/ml,硝普钠盐含量为1 mg/ml;苯酚试剂中苯酚含量为50 mg/ml,NaOH含量为21.... 详细信息

采用正交实验对显色试剂配比进行优化,并探讨了水质分析理化条件,确定了现场分析氨氮的靛酚蓝法.其结果表明:复合试剂中柠檬酸钠含量为120 mg/ml,EDTA含量为30 mg/ml,硝普钠盐含量为1 mg/ml;苯酚试剂中苯酚含量为50 mg/ml,NaOH含量为21.2 mg/ml;DCI试剂中DCI含量为20 mg/ml,NaOH含量为14.8 mg/ml时可以缩短反应时间在10 min内,具有快速、操作简便等优点,测定结果的精密度和准确度,均能满足海上现场分析的要求.

关键词：水质分析靛酚蓝法正交实验显色试剂配比

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荧光法测定海洋生物体中石油烃总量

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大连海事大学学报 2006年第1期32卷 51-53页

作者：王倩邵秘华陶平大连海事大学环境科学与工程学院辽宁大连116026

将生物样品经氢氧化钠皂化，采用二氯甲烷萃取，残留物用石油醚溶解，采用荧光分光光度计，于激发波长310nm，发射波长360nm处进行总量测定．分析结果表明，方法的检测下限值为1．89μg／L。重复性较好．样品分析结果精密度，其相对误... 详细信息

将生物样品经氢氧化钠皂化，采用二氯甲烷萃取，残留物用石油醚溶解，采用荧光分光光度计，于激发波长310nm，发射波长360nm处进行总量测定．分析结果表明，方法的检测下限值为1．89μg／L。重复性较好．样品分析结果精密度，其相对误差不超出8％，准确度内控样加标，回收率测定结果在86％～102％之间．

关键词：荧光分光光度法生物体石油烃二氯甲烷

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Recombinant human growth hormone secreted from tissue-engineered bioartificial muscle improves left ventricular function in rat with acute myocardial infarction

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Chinese Medical Journal 2009年第19期122卷 2352-2359页

作者： RONG Shu-ling WANG Yong-jin WANG Xiao-lin LU Yong-xin CHANG Chao WANG Feng-zhi LIU Qi-yun LIU Xiang-yang GAO Yan-zhang mi shao-hua Department of Cardiology Heping Hospital Changzhi Medical College Changzhi Shanxi 046000 China Department of Paediatrics Heping Hospital Changzhi Medical College Changzhi Shanxi 046000 China Department of Cardiology Union Hospital Tongji Medical College Huazhong University of Science and Tecnology Wuhan Hubei 430022 China Department of Cardiology Second Hospital of Shanxi Medical University Taiyuan Shanxi 030001 China

Background Experimental studies and preliminary clinical studies have suggested that growth hormone （GH） treatment may improve cardiovascular parameters in chronic heart failure （CHF）. Recombinant human GH （rhGH） has been delivered by a recombinant protein, by plasmid DNA, and by genetically engineered cells with different pharmacokinetic and physiological properties. The present study aimed to examine a new method for delivery of rhGH using genetically modified bioartificial muscles （BAMs）, and investigate whether the rhGH delivered by this technique improves left ventricular （LV） function in rats with CHE Methods Primary skeletal myoblasts were isolated from several Sprague-Dawley （SD） rats, cultured, purified, and retrovirally transduced to synthesize and secrete human rhGH, and tissue-engineered into implantable BAMs. Ligation of the left coronary artery or sham operation was performed. The rats that underwent ligation were randomly assigned to 2 groups： CHF control group （n=6） and CHF treatment group （n=6）. The CHF control group received non-rhGH-secreting BAM （GFP-BAMs） transplantation, and the CHF treatment group received rhGH-secreting BAM （GH-BAMs） transplantation. Another group of rats served as the sham operation group, which was also randomly assigned to 2 subgroups： sham control group （n=6） and sham treatment group （n=6）. The sham control group underwent GFP-BAM transplantation, and the sham treatment group underwent GH-BAM transplantation. GH-BAMs and GFP-BAMs were implanted subcutaneously into syngeneic rats with ligation of the left coronary artery or sham operation was performed. Eight weeks after the treatment, echocardiography was performed, hGH, insulin-like growth factor-1 （IGF-1） and TNF-a levels in rat serum were measured by radioimmunoassay and ELISA, and then the rats were killed and ventricular samples were subjected to immunohistochemistry. Results Primary rat myoblasts were retrovirally transduced to secrete rhGH and tissue-engineered int

关键词： heart failure muscles growth hormone gene therapy

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船载雷达观测海浪的研究

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海洋学报 2006年第5期28卷 152-156页

作者：任福安邵秘华孙延维大连海事大学轮机工程学院辽宁大连116026 大连海事大学环境科学与工程学院辽宁大连116026

1引言随着海洋工程的迅速发展,海洋资源开发和海洋环境保护以及海洋减灾防灾监测预报服务等各方面都特别需要沿岸近海的海浪动态特征信息.目前对海浪研究一般采用两种方法,一是动力学方法,二是统计学方法,但在研究中往往将两种方法结合... 详细信息

1引言随着海洋工程的迅速发展,海洋资源开发和海洋环境保护以及海洋减灾防灾监测预报服务等各方面都特别需要沿岸近海的海浪动态特征信息.目前对海浪研究一般采用两种方法,一是动力学方法,二是统计学方法,但在研究中往往将两种方法结合起来[1].比较经典的方法是利用流体力学方程研究理想的规则波动(正弦波动、余弦波动、斯托克斯波动等),把海水视为不可压缩的理想流体,运动是无旋的.这一类的研究成果能够说明海洋中实际发生的一些比较规则的波动现象.另一种研究海浪的方法是将实际观测资料与波动理论结合起来,将实际海浪视为许许多多振幅不等、相位各不相同的正弦波的叠加,利用谱分析方法确定组成波谱的特征,但是对海浪观测,尤

关键词：船载雷达图像波谱海浪

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基于ECDIS的雷达遥测海冰监控系统

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中国造船 2008年第2期49卷 107-112页

作者：赵宝刚郝江凌邵秘华大连海事大学环境科学与工程学院辽宁大连116026 大连海事大学航海学院辽宁大连116026

依据渤海辽东湾海冰管理项目,将电子海图显示与信息系统(ECDIS)和雷达海冰监控相结合,构建了一种雷达遥测监控系统。陆地雷达监控站使用MD-3641/3642型雷达,采用Internet技术和CDMA/GPRS通信技术实现与指挥中心的数据传输。设计的嵌入... 详细信息

依据渤海辽东湾海冰管理项目,将电子海图显示与信息系统(ECDIS)和雷达海冰监控相结合,构建了一种雷达遥测监控系统。陆地雷达监控站使用MD-3641/3642型雷达,采用Internet技术和CDMA/GPRS通信技术实现与指挥中心的数据传输。设计的嵌入式雷达控制单元以16位单片机PIC24作为CPU,并给出一种目标回波识别算法,用以实现图像去噪和确定海冰回波的准确位置与分布。网站结果表明,该系统扩展了雷达遥测海冰及监控的功能,对保护海洋石油平台和海上工程设施及航运的安全预警乃是一种有效管理途径。

关键词：船舶舰船工程雷达遥测海冰电子海图显示与信息系统回波识别算法

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近岸海域多环芳烃生态系统动力学模型及生境影响

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中国环境科学 2016年第5期36卷 1540-1546页

作者：张晓霞程嘉熠陶平邵秘华朱京海大连海事大学环境科学与工程学院辽宁大连116000 辽宁省环境保护厅辽宁沈阳110000

利用生态动力学模型Delf3D,对大连葫芦山湾海域多环芳烃(PAHs)在海洋中的理化及生态动力学过程进行耦合,并以大连葫芦山湾实际监测数据及相关文献资料为基础,确定了一套适用于该近岸海域PAHs生态动力学过程的特征参数,模拟研究了PAHs在... 详细信息

利用生态动力学模型Delf3D,对大连葫芦山湾海域多环芳烃(PAHs)在海洋中的理化及生态动力学过程进行耦合,并以大连葫芦山湾实际监测数据及相关文献资料为基础,确定了一套适用于该近岸海域PAHs生态动力学过程的特征参数,模拟研究了PAHs在近岸海域生态动力学过程.以浮游植物生长率作为指示近岸海域生态系统健康程度的指标,将PAHs含量的数值模拟结果与浮游植物生长率的动力学响应方程相拟合,定量的评估了PAHs对海洋生态系统的影响.结果显示:排污口选址于水深大、流速快、水动力强处,对PAHs污染物扩散、水质改善具有显著作用,反之造成PAHs大规模富集.PAHs富集作用对该湾生境影响较显著,湾顶水动力作用微弱,浮游生物生长率削减度高达18%.Arc GIS分析显示,浮游生物生长率削减程度与PAHs时空分布并不完全一致,体现海湾生态系统的动力复杂性与结构稳固性.

关键词：近岸海域生态动力学模型多环芳烃数值模拟 Delft3D

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