血管内皮细胞生长因子基因治疗闭塞性血管病研究进展

概述了近年来利用血管内皮细胞生长因子（ＶＥＧＦ）基因治疗闭塞性血管病的成果,初步探讨了ＶＥＧＦ基因的临床疗效及其应用的安全性。     

基因枪技术及其在基因治疗中的应用进展

目前基因转染载体主要分为病毒型载体和非病毒型载体,病毒载体虽转染率较高、表达时间长,但其安全性令人担忧,非病毒载体中基因枪的优势最为明显,临床化趋势最强。通过分析非病毒基因转染技术面临的障碍,介绍了基因枪技术的产生和原理及其显著的优势,并总结了当前基因枪技术在基因治疗中的应用,指出了基因枪技术发展面临的问题和发展方向。     

基因枪在RNA干扰中的应用研究进展

RNA干扰（RNA interference,RNAi）是指双链RNA（double-strand RNA,dsRNA）特异性降解同源mRNA,从而引发基因转录后水平沉默的现象,是一种高效、高特异性抑制基因表达的途径。自1998年Fire等发现RNA干扰现象以来,其特异性降解目的基因的优势吸引了众多研究者的目光。本文在简要综述RNAi技术在基因功能研究、抗病毒治疗,肿瘤基因治疗等领域的应用后,重点归纳了基因枪技术在RNAi研究即siRNA导入细胞中的应用,并简单分析其优势与意义。     

高速微粒基因枪在植物基因工程中的应用

人口的过度增长。耕地面积的日益减少,国民经济的发展和人民生活水平的逐步改善,迫切要求提高作物产量和产品质量。传统的育种方法虽然还有不不可低估的潜力,但由于种质资源的贫乏和生物种间的差异限制,就显示出不可回避的局限性。这样,使得人们不得不去寻找有效的改良作物品种的新方法、新技术来缓解这一矛盾。以改良作物创造新品种作为战略目标的植物基因工程的兴起,向人们展示了崭新的农业分子育种方法。植物基因工程不仅要对目的基因进行鉴定、分离和克隆,更重要的是要将     

肝纤维化基因治疗的进展

肝硬化是慢性肝病晚期的组织学改变 ,以纤维组织大量增生和肝小叶结构无序化为特征 ,因此又称肝纤维化。近年来随着分子生物学的发展 ,肝纤维化的分子机制逐渐得以阐明 ,从而使肝纤维化的基因治疗成为可能。肝纤维化的基因治疗主要起到阻止纤维化发展、刺激肝细胞分裂和肝组织结构重建三方面的作用。目前 ,常用的方法一般是通过缺陷病毒 (如腺病毒 )转入特定的细胞因子和酶 (如肝细胞生长因子、转化生长因子β1受体、基质金属蛋白酶等 )的基因 ,通过靶细胞表达这些因子作用于受损的肝脏 ,达到延缓和治愈肝纤维化的目的     

基因枪在植物遗传转化中的应用

基因枪技术是近年发展起来的一项新的植物遗传转化技术,本文较为系统地介绍了基因枪的类型,以及它们各自的特点和工作原理,并对基因枪在植物遗传转化中的广泛用途作了重点介绍,文中引用用参考文献七十余篇,为进一步深入了解学科的最新研究进展提供了方便。     

miRNAs在肝细胞肝癌基因治疗中的研究进展

肝癌是严重威胁人类健康的主要恶性肿瘤之一,其中70%~85%是肝细胞肝癌(hepatocellular carcinoma,HCC)。目前,HCC的各种治疗方法的效果均具局限性。HCC发生发展中的各种生物大分子的变化及其机制是探索新的有效治疗方法的基础。近二十年来发现的微小RNA(micro RNA或mi RNA)在基因表达、蛋白质翻译过程中发挥着重要的调控作用,其异常表达与肿瘤的发生发展密切相关。与此同时,基因治疗作为一种新兴的生物治疗手段,也是目前的研究热点之一。因此,mi RNA正被应用于肿瘤的基因治疗研究之中。该文就mi RNA的作用机制、mi RNA与HCC的关系和mi RNA在HCC基因治疗中的应用研究作一综述。     

基因治疗在慢性难愈创面的应用进展

众多疾病和创伤都会带来损伤创面的难愈问题,慢性难愈创面的长期存在不仅增加了局部感染的机会,而且大大影响了局部组织的结构和功能。基因治疗作为一种新的生物治疗方法,其相关理论和技术近来日臻完善,为慢性难愈创面的治疗提供了新的技术途径,有望带来突破性的治疗效果。     

基因枪技术在植物基因工程研究中的应用

文章介绍了近年来采用基因枪技术在植物基因工程研究中开展的一些工作。     

基因枪在水稻遗传转化中的应用及其转化技术的优化

1983年Zambryski等人用根瘤农杆菌介导法进行烟草基因转移,获得了世界上首例转基因植株.随后,应用DNA直接导入技术如电击法(electroporation)和PEG介导法(PEG—mediated)成功地获得了转基因水稻植株.近年来,随着基因枪技术的建立和发展,水稻遗传转化成功的报道逐年增多.目前基因枪技术在植物遗传转化中的应用超过了根瘤农杆菌介导和其它转化方法的应用.这是因为基因枪转化技术不受植物种类的限制,不需要以原生质体作为转化的受体,可以将外源基因直接导入细胞、组织或器官,因而克服了根瘤农杆菌     

Marked inhibition of retinal neovascularization in rats following soluble-flt-1 gene transfer

BACKGROUND: In mouse models of retinopathy of prematurity (ROP) inhibitors of vascular endothelial growth factor (VEGF) functions administered systemically completely block retinal neovascularization. In contrast, selective ocular VEGF depletion has achieved an approx. 50% inhibition of retinal neovascular growth. It is unclear whether a more complete inhibition of new blood vessel development can be obtained with an anti-VEGF therapy localized to the eye. Therefore, the objective of the present study was to determine the effect of local anti-VEGF therapy in a different animal model which closely mimics human ROP. METHODS: Rats were exposed to alternating cycles of high and low levels of oxygen for 14 days immediately after birth; thereafter, they were intravitreally injected with an adenoviral vector expressing a secreted form of the VEGF receptor flt-1 (Ad.sflt), which acts by sequestering VEGF. Contralateral eyes were injected with the control vector carrying the reporter gene expressing beta-galactosidase (Ad.betaGal). RESULTS: At the peak of retinal neovascular growth, i.e. post-natal day 21 (P21), we observed up to 97.5% decrease in retinal neovascularization in animals injected with Ad.sflt. At the end of observation (P28), no significant difference in retinal vessel number was detected in both oxygen-injured and normoxic Ad.sflt-treated retinas compared with untreated or Ad.betaGal-treated retinas. CONCLUSION: Adenoviral-mediated sflt-1 gene transfer induces a near-complete inhibition of ischemia-induced retinal neovascularization in rats without affecting pre-existing retinal vessels.     

农杆菌及基因枪在玉米遗传转化中的应用

目前外源基因导入玉米受体细胞的方法很多 ,最受瞩目的是农杆菌介导法和基因枪法。就农杆菌、基因枪转化玉米的基本原理、影响转化率的因素 ,以及近年来在玉米遗传转化中的最新动态进行了综述     

质粒治疗肢体动脉闭塞性疾病的安全性分析

肢体动脉血管闭塞性病是一种严重危害人类健康的疾病,许多病人最终要截肢,基因治疗为这类患带来了希望。利用肝细胞生长因子促进血管生成的作用,采用质粒作为载体对大鼠和故事片诉下肢动脉闭塞疾病模型进行治疗,效果良好。为探讨基因治疗的安全性问题,用绿色荧光蛋白报告基因研究质粒在体内的分布和表达,方法简便。通过制作冰冻切片观察绿色荧光蛋白在小鼠体内的表达情况发现局部注射质粒不会造成全身性扩散,表明该方法安全。     

VEGF 164 gene transfer by electroporation improves diabetic sensory neuropathy in mice

BACKGROUND: Diabetic neuropathy is the most common cause of peripheral neuropathy and a serious complication of diabetes. Vascular endothelial growth factor (VEGF) stimulates angiogenesis and has neurotrophic and neuroprotective activities. To examine the efficiency of VEGF 164 electro-gene therapy for neuropathy, intramuscular VEGF 164 gene transfer by electroporation was performed to treat sensory neuropathy in diabetic mice. METHODS: VEGF 164 was overexpressed in the tibial anterior (TA) muscles of streptozotocin-induced diabetic mice with hypoalgesia, using a VEGF 164 plasmid injection with electroporation. From 2 weeks after electro-gene transfer, the nociceptive threshold was measured weekly using the paw-pressure test. The TA muscles, sciatic nerve, liver and spleen were histochemically examined at 4 weeks after electro-gene transfer. RESULTS: Two weeks after electro-gene transfer into the bilateral TA muscles, the elevated nociceptive threshold was decreased to a normal level in all treated mice. Improvement of the hypoalgesia continued for 14 weeks. When the VEGF 164 plasmid was injected with electroporation into a unilateral TA muscle, recovery from hypoalgesia was observed in not only the ipsilateral hindpaw, but also the contralateral one, suggesting that VEGF circulates in the blood. No increase in the number of endoneurial vessels in the sciatic nerve was found in the VEGF 164 plasmid-electroporated mice. CONCLUSIONS: These findings suggest that VEGF 164 electro-gene therapy completely recovered the sensory deficits, i.e. hypoalgesia, in the diabetic mice through mechanisms other than angiogenesis in the endoneurium of the peripheral nerve, and may be useful for treatment for diabetic sensory neuropathy in human subjects.     

Treatment of chronical myocardial ischemia by adenovirus-mediated hepatocyte growth factor gene transfer in minipigs

Growth factor gene transfer-induced therapeutic angiogenesis has become a novel approach for the treatment of myocardial ischemia. In order to provide a basis for the clinical application of an adeno- virus with hepatocyte growth factor gene (Ad-HGF) in the treatment of myocardial ischemia, we estab- lished a minipig model of chronically ischemic myocardium in which an Ameroid constrictor was placed around the left circumflex branch of the coronary artery (LCX). A total of 18 minipigs were ran- domly divided into 3 groups: a surgery control group, a model group and an Ad-HGF treatment group implanted with Ameroid constrictor. Ad-HGF or the control agent was injected directly into the ischemic myocardium, and an improvement in heart function and blood supply were evaluated. The results showed that myocardial perfusion remarkably improved in the Ad-HGF group compared with that in both the control and model groups. Four weeks after the treatment, the density of newly formed blood vessels was higher and the number of collateral blood vessels was greater in the Ad-HGF group than in the model group. The area of myocardial ischemia reduced evidently and the left ventricular ejection fraction improved significantly in the Ad-HGF group. These results suggest that HGF gene therapy may become a novel approach in the treatment of chronically ischemic myocardium.     

肝细胞生长因子对四氯化碳损伤肝细胞的保护作用及相关基因表达

利用无血清原代培养大鼠肝细胞,观察重组人肝细胞生长因子（ｒｈＨＧＦ）对ＣＣｌ４染毒肝细胞的保护作用。结果表明：（１）ｒｈＨＧＦ（５ｎｇ／ｍｌ）预自理后可显著提高ＣＣｌ４（１５ｍｍｏｌ／Ｌ）染毒肝细胞存活率,降低细胞内丙氨酸氨基转移酶（ＡＬＴ）、Ｋ＾＋的漏出;（２）表皮生长因子（ＥＧＦ,５０ｎｇ／ｍｌ）和ｒｈＨＧＦ（５ｎｇ／ｍｌ）合用预处理肝细胞,ＣＣｌ４染毒后细胞内ＡＬＴ、Ｋ＾＋漏出较ｒｈＨＧＦ和