pDR2/TK基因经肝动脉注射后杀伤效应的研究

目的 探讨ｐＤＲ２／ＴＫ基因经大鼠肝左叶动脉注射后的靶向性表达及杀伤效应。方法 将４８只大鼠随机分为４组,分别给予ｐＤＲ２／ＴＫ基因、更革洛韦（ｇａｎｃｉｃｏｖｉｒ,ＧｃＶ）、ｐＤＲ２／ＴＫ基因／生理盐水（ＮＳ）、缓冲液（ＴＥ）／ＧＣＶ及ＴＥ／ＧＣＶ。肝组织用原位杂交、免疫组织化学ＡＢＣ法及苏木素－伊红（ＨＥ）染色的方法检测基因转录、表达及杀伤效应。结果 ｐＤＲ２／ＴＫ基因在肝左叶有转录。单独注射     

前列腺素E_1、人胎盘神经生长因子对周围神经损伤后运动恢复的作用

目的 探讨前列腺素Ｅ１（ＰＧＥ１）、人胎盘种经生长因子（ＨＮＧＦ）对周围神经损伤后运动功能恢复的影响。方法 取大鼠 ３５只,作坐骨神经夹毁模型,术侧分别注射 ＰＧＥ１、ＨＮＧＦ１４日,用三导记录肌肉收缩张力,术后每日测术侧和正常侧足趾伸张距离的比值（ＥＴＳ／ＮＴＳ）及足印长度的均值（ＥＰＬ／ＮＰＬ）,并分别与生理盐水组及无神经损伤组进行比较研究。结果ＰＧＥ１和ＨＮＧＦ能明显缩短足趾伸张距离和足印长度的恢复时间,明显增进肌肉收缩力、加快收缩峰值速度和减轻肌肉萎缩率,但ＰＧＥ１的作用效果低于 ＨＮＧＦ结论 ＨＮＧＦ可能通过某些间接作用途径促进神经损伤后运动功能恢复;ＰＧＥ１能部分模拟 ＮＧＦ促进运动功能恢复的作用。     

移植肾急性排斥时血栓素A2、前列环素代谢改变及其对肾小球滤过率的影响

用放射免疫法检测２１例移植肾急性排斥时，肾移植患者尿液以及血浆血栓素Ｂ２（ＴＸＢ２）和６－酮－前列腺素Ｆ１α（６－ｋｅｔｏ－ＰＧＦ１α）浓度，并检测４例不可逆急性排斥和７例慢性排斥移植肾切除后肾组织ＴＸＢ２和６－ｋｅｔｏ－ＰＧＦ１α含量。发现急性排斥出现时，尿中ＴＸＢ２和６－ｋｅｔｏ－ＰＧＦ１α含量均明显升高，ＴＸＢ２增多出现较早；血浆ＴＸＢ２浓度也显著增加，６－ｋｅｔｏ－ＰＧＦ１α浓度下降。尿液和血浆中ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１α比值增大。正常肾组织标本中，肾髓质ＴＸＢ２和６－ｋｅｔｏ－ＰＧＦ１α含量为皮质的４～５倍，皮髓质中ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１α比值水平相同。不可逆急性排斥肾组织中，肾皮质ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１α比值明显高于髓质。急性排斥时，尿液中ＴＸＢ２和６－ｋｅｔｏ－ＰＧＦ１α比值变化与肾皮质平行。分析表明，急性排斥时，尿液ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１α比值增大和移植肾肾小球滤过率负相关。     

肢体缺血再灌注导致脂质过氧化增强和血栓素A2合成增加的研究

骨骼肌缺血再灌注可导致肌肉微血管的实质损伤，观察了肌肉缺血再灌注时脂质过氧化物（ＬＰＯ）和ＴｘＡ２与ＰＧＩ２的代谢状况。缺血组家兔（ｎ＝６）。双后肢缺血２小时。再灌注２小时后，左股薄肌丙二醛（ＭＤＡ）含量，下腔静脉血ＭＤＡ和乳酸浓度均显著高于对照组（ｎ＝６）。再灌注后１０和３０分钟，缺血组血中ＴｘＢ２水平和ＴｘＢ２／６－ｋｅｔｏ－ＰＧＦ１ａ比率明显增高，再灌注后血中ＳＯＤ活性和６－ｋｅｔｏ－ＰＧＦ     

再灌注损伤胃粘膜组织前列腺素含量的变化及丹参提取物F的作用

研究失血性休克再灌注大鼠胃粘膜损伤指数，组织 ＰＧＥ２、 ＰＧＩ２（ ６－ｋｅｔｏ－ＰＧＦ１ａ示） ＴＸＡ２（ＴＸＢ２示）含量，６－ｋｅｔｏ－ＰＧＦ１ａ／ＴＸＢ２比值及丹参提取物 Ｆ对上述各值的影响。结果显示，再灌注后胃粘膜有明显的出血性损伤，组织ＰＧＥ２、６－ｋｅｔｏ－ＰＧＦ１ａ含量明显下降，ＴＸＢ２含量明显增加，６－ｋｅｔｏ－ＰＧＦ１ａ／ＴＸＢ２比值明显降低，单纯失血性休克组未见上述损伤；丹参提取物Ｆ能明显降低胃粘膜损伤指数，且与组织ＰＧＥ２含量６－ｋｅｔｏ－ＰＧ１ａ／ＴＸＢ２比值的增加呈负相关。     

体外震波碎石前后血浆和尿液PGI2与TXA2的变化及其临床意义

应用放射免疫技术，对１４例非梗阻性单纯肾结石患者ＳＥＷＬ前后血浆和尿液中ＴＸＢ２和６－ｋｅｔｏ－ＰＧＦ１ａ水平进行了动态检测。结果显示，血浆ＴＸＢ２在ＥＳＷＬ后第一天明显升高（Ｐ＜０．００１），而６－ｋｅｔｏ－ＰＧＦ１α无显著意义的改变（Ｐ＞０．０５），两者的比值也随ＴＸＢ２而升高（Ｐ＜Ｏ．００１），第三天时都降至基础水平；尿液中上述指标也出现类似变化。我们认为，ＥＳＷＬ后早期肾脏内ＴＸＡ２合成与释放增加，ＴＸＡ２与ＰＧＩ２平衡失调，有可能参与了肾损害的病理生理过程。     

心脏停跳复苏后全脑浅低温对脑组织前列环素和血栓素A2含量…

观察犬心脏停跳复跳后全脑浅低温对脑组织前列环素和血栓素Ａ２含量的影响。结果表明，脑缺血再灌注后常规治疗４小时，６－ｋｅｔｏ－ＰＧＦ１ａ水平无明显变化。ＴＸＢ２含量明显上升，ＴＸＢ２／６－ｋｔｅｏ－ＰＧＦ１ａ（Ｔ／Ｋ）比值明显升高；浅低温治疗组与缺血再灌常规治疗组比较６－ｋｅｔｏ－ＰＧＦ１ａ水平仍无明显变化，而ＴＸＢ２含量下降（Ｐ＜０．０５），Ｔ／Ｋ比值降低（Ｐ＜０．０５）。提示全脑浅低温能抑制脑缺     

小儿先天肾发育不全和畸形103例临床病理分析

小儿先天肾发育不全和畸形１０３例临床病理分析曾丽霞，高岩ＣＨＩＬＤＨＯＯＤＣＯＮＧＥＮＩＴＡＬＲＥＮＡＬＨＹＰ－ＯＰＬＡＳＩＡＡＶＤＡＩＯＭＡＬＹＣＬＩＮＩＣＡＬＡＮＤＰＡ－ＴＨＯＬＯＧＩＣＡＬＳＴＬＵＤＹＯＦ１０３ＣＡＳＥＳＺｅｎｇＬｉｘｉａ，Ｇａ...     

阿魏酸钠对犬心脏停跳复苏后脑组织TXB_2、6-keto-PGF_(1a)及MDA含量的影响

研究了阿魏酸钠对犬心脏停跳１０分钟复苏后４小时脑组织中血栓素Ｂ２（ＴＸＢ２）、６－酮－前列腺素Ｆ１ａ（６－ｋｅｔｏ－ＰＧＦ１ａ）及丙二醛（ＭＤＡ）含量的影响。１７只犬随机分为非缺血对照组（Ａ组）、缺血再灌注常规治疗组（Ｂ组）及缺血再灌注阿魏酸钠治疗组（Ｃ组）。结果发现，Ｂ组ＴＸＢ２、ＭＤＡ含量及ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１ａ比值均较 Ａ组明显升高（Ｐ＜０． ０１）。 Ｃ组 ＴＸＢ２、ＭＤＡ含量及 ＴＸＢ２／６－ｋｅｔｏ－ＰＧＦ１ａ比值升高幅度均较Ｂ组明显低（Ｐ＜０．０１）。表明阿魏酸钠可明显抑制犬心脏停跳复苏后脑组织花生四烯酸代谢及脂质过氧化反应。     

大鼠脊髓损伤后兴奋性氨基酸的变化及其对血流量的影响

采用Ａｌｌｅｎ＇ｓ打击法复制大鼠脊髓损伤（ＳＣＩ）模型，氨基酸微量检测技术和氢清除法分别测定伤段脊髓组织２４ｈ内兴奋性氨基酸（ＥＡＡ）即谷氨酸（Ｇｌｕ）、天冬氨酸（Ａｓｐ）含量和脊髓灰质血流量（ＳＣＢＦ）的变化，并观察脊髓蛛网膜下腔注射ＥＡＡ受体激动剂Ｎ－甲基－Ｄ－天冬氨酸（ＮＭＤＡ）对ＳＣＢＦ的影响，探讨ＥＡＡ在ＳＣＩ中的作用。结果发现：ＳＣＢＦ在伤后１０ｍｉｎ即有明显下降，２ｈ较１ｈ略有回升，４～８ｈ又进一步下降，第二次下降与第一次下降相差显著。伤后脊髓蛛网膜下腔注射ＮＭＤＡ明显加剧ＳＣＩ后脊髓缺血。Ｇｌｕ、Ａｓｐ伤后１０ｍｉｎ均明显升高，１～２４ｈＡｓｐ较对照组明显降低，８ｈ较２ｈ略有回升；Ｇｌｕ在伤后１０ｍｉｎ有所下降，但仍高于对照组，４ｈ、８ｈ较２ｈ略增加。ＥＡＡ变化与ＳＣＢＦ呈显著负相关。结果提示ＳＣＩ后ＥＡＡ的过度释放是ＳＣＩ后继发损伤的重要因素。     

自体骨髓干细胞移植治疗下肢缺血的研究

目的　为临床探索一种更简便安全的下肢动脉缺血性疾病的治疗途径。方法　建立鼠后肢缺血动物模型 ,将取于自体的骨髓细胞制成悬液注射于缺血部位 ,4周后行动脉造影并取标本 ,测定毛细血管密度。结果　动脉造影显示大鼠缺血肢体侧支动脉明显增多 ,毛细血管密度增加。结论　自体骨髓干细胞移植可望成为一种简单的、有效的治疗下肢缺血的方法     

激光心肌血运重建术联合血管内皮细胞生长165基因治疗缺血心肌的研究

目的：观察激光心肌血运重建术（TMLR）联合血管内皮细胞生长因子165基因（VEGF165cDNA）治疗缺血心肌后心肌血管密度、心肌灌注和代谢的变化。方法：健康杂种犬48条,随机分为TMLR组、TMLR联合VEGF165cDNA组和空质粒（pcDNA3.1）组（n=12）,心肌梗死为对照组。结扎冠状动脉致心肌缺血60min后行CO2激光心肌打孔,VEGF165cDNA基因转染采用直接心肌注射法。结果：TMLR联合VEGF165基因治疗后6和12周,缺血区心肌血管密度和心肌灌注量、代谢显著高于TMLR组。结论：TMLR联合VEGF165基因治疗缺血心肌后心肌血管密度显著增加;成活心肌的数量显著增加和/或成活心肌功能显著恢复。     

Enhanced brain angiogenesis in chronic cerebral hypoperfusion after administration of plasmid human vascular endothelial growth factor in combination with indirect vasoreconstructive surgery

OBJECT: Vascular endothelial growth factor (VEGF) is a secreted mitogen associated with angiogenesis. The conceptual basis for therapeutic angiogenesis after plasmid human VEGF gene (phVEGF) transfer has been established in patients presenting with limb ischemia and myocardial infarction. The authors hypothesized that overexpression of VEGF using a gene transfer method combined with indirect vasoreconstruction might induce effective brain angiogenesis in chronic cerebral hypoperfusion, leading to prevention of ischemic attacks. METHODS: A chronic cerebral hypoperfusion model induced by permanent ligation of both common carotid arteries in rats was used in this investigation. Seven days after induction of cerebral hypoperfusion, encephalomyosynangiosis (EMS) and phVEGF administration in the temporal muscle were performed. Fourteen days after treatment, the VEGF gene therapy group displayed numbers and areas of capillary vessels in temporal muscles that were 2.2 and 2.5 times greater, respectively, in comparison with the control group. In the brain, the number and area of capillary vessels in the group treated with the VEGF gene were 1.5 and 1.8 times greater, respectively, relative to the control group. CONCLUSIONS: In rat models of chronic cerebral hypoperfusion, administration of phVEGF combined with indirect vasoreconstructive surgery significantly increased capillary density in the brain. The authors' results indicate that administration of phVEGF may be an effective therapy in patients with chronic cerebral hypoperfusion, such as those with moyamoya disease.     

纳米血管内皮生长因子在治疗下肢缺血促进血管再生成中的作用

目的　利用家兔后肢缺血模型 ,观察纳米材料聚乳酸聚乙醇酸共聚物 (poly dl lactic co glycolicacid ,PLGA) ,包载血管内皮生长因子 (VEGF16 5 )基因 ,经局部肌肉注射后 ,外源基因在局部缺血组织中的转染及强度 ,以及缺血部位的血管新生状况。　方法　制备VEGF16 5 真核表达质粒 ,制备包载VEGF16 5 基因的纳米粒子 ,并检测其理化性质和体外释放曲线。建立家兔后肢缺血模型 2 4只 ,其中4只为对照组 ,只行股动脉及其分支结扎切断术 ;2只为空白纳米粒子组 ,局部缺血肌肉注射空白纳米粒子 ;10只应用裸质粒VEGF16 5 转染 ,8只采用纳米技术包载VEGF16 5 转染。直接缺血部位肌肉内多点注射 ,进行局部定位基因转染。术后 14d行血管造影 ,了解缺血部位侧枝形成情况。处死兔子 ,取股二头肌 ,内收大肌 ,做病理切片 ,免疫组化染色 ,观察VEGF16 5 的表达 ,测定毛细血管密度。应用逆转录 聚合酶链反应了解VEGF16 5 在骨骼肌中的表达 ,并对不同的转染技术进行半定量分析。　结果　转基因治疗 14d后 ,转染VEGF基因组血管造影可见明显新生血管和侧枝循环形成 ,免疫组化染色可见VEGF16 5 蛋白表达水平增高 ,缺血肌肉血管数增多 ,纳米VEGF16 5 治疗组与裸质粒VEGF16 5 治疗组的毛细血管密度明显高于对照组 ,有显著性差异 (P     

Vascular endothelial growth factor gene delivery by magnetic DNA nanospheres ameliorates limb ischemia in rabbits

BACKGROUND: Critical limb ischemia often leads to disability and limb loss. Vascular endothelial growth factor (VEGF), delivered either as recombinant protein or as gene therapy, has been shown to promote arteriogenesis and angiogenesis in animal models of limb ischemia. However, most of the studies used a nonspecific targeting system. MATERIALS AND METHODS: Magnetic DNA nanospheres containing expression plasmids encoding VEGF were synthesized, and their morphology, magnetropism, and stability were analyzed. The magnetic DNA nanospheres were administrated via an artery into a rabbit limb ischemia model. The expression of VEGF and vascularization were examined by immunohistochemistry. The angiography was taken to evaluate arteriogenesis. RESULTS: Magnetic DNA nanospheres were very stable and showed a high magnetropism. Gene delivery of such nanospheres via artery under a magnetic field led to the overexpression of VEGF in situ. The capillary density and capillary to muscle fiber ratio were doubled compared with those of the control animals. The arteriogenesis also was promoted in VEGF gene therapy group compared with controls but at later interval than capillary angiogenesis. CONCLUSIONS: Our results suggest that intra-arterial VEGF gene delivery by magnetic DNA nanosphere promotes angiogenesis and arteriogenesis and presents a potent therapeutic strategy for critical limb ischemia.     

人血管内皮细胞生长因子基因转染大鼠骨髓间充质干细胞对脂肪颗粒移植存活率的影响

目的 检测大鼠骨髓间充质干细胞(BMSCs)经人血管内皮细胞生长因子(VEGF165)基因转染后,对脂肪颗粒移植存活率的影响.方法　利用FuGENE HD介导VEGF165基因体外转染SD大鼠BMSCs,与取自SD大鼠的脂肪组织混合后移植于大鼠背部,类似方法设立未转染组及DMEM液空白对照组.检测脂肪组织存活率及再生血管密度.结果　转染组存在外源性基因和蛋白的表达,脂肪组织存活率及再生血管密度高于未转染组,且两者均高于对照组.结论　转染VEGF165基因的BMSCs具有更强的促血管再生作用,可提高游离移植脂肪的存活率.     

转染血管内皮生长因子基因的小鼠NIH3T3细胞移植促进皮瓣早期血运重建的研究

目的探讨转染血管内皮生长因子（VEGF）基因的小鼠NIH3T3细胞移植对缺血皮瓣的血管新生和皮瓣存活率的影响。方法体外PcDNA3．1（-）／VEGF165质粒转染小鼠NIH3T3细胞,免疫组化方法检测小鼠NIH3T3细胞体外表达VEGF的情况,CM-DiI标记小鼠NIH3T3细胞。将小鼠随机分为3组：A组[PcDNA3．1（-）／VEGF165质粒转染的NIH3T3细胞移植]、B组（单纯NIH3T3细胞移植）、C组（单纯DMEM培养基注射）。每只小鼠背侧皮下按组分别注射细胞悬液和培养基,注射后酶联免疫吸附（ELISA）法连续检测大鼠血浆VEGF浓度,注射后第4天掀起一个蒂在尾侧的4．0cm×1．5cm的随意皮瓣。术后第7天分别观察皮瓣的存活率、血流灌注、皮瓣毛细血管密度、NIH3T3细胞在皮瓣内的分布和存活情况。结果转染VEGF165基因的小鼠NIH3T3细胞体外和体内检测均高表达VEGF165蛋白。A组的皮瓣存活率、毛细血管密度、血流灌注比值均显著高于另外两组（P〈0．05）。结论转染VEGF基因的小鼠NIH3T3细胞皮下移植可促进缺血皮瓣的血管新生,提高存活率。     

基因修饰的骨髓间充质干细胞移植于慢性心肌梗死模型的实验研究

目的 研究血管内皮生长因子（VEGF）165基因修饰的骨髓间充质干细胞（MSCs）移植于慢性心肌梗死模型后的血管新生及对心功能的作用。方法 同源重组法构建含有VEGF。基因的重组腺病毒载体（rAd—VEGF165）;密度梯度离心法分离骨髓单个核细胞,贴壁法培养兔MSCs;rAd—VFGF165转染MSCs,并用4,6-联脒-2-苯基吲哚（DAPI）标记MSCs;结扎前降支法建立兔心肌梗死模型,存活6周的实验动物36只随机分为3组：移植rAd—VFGF165转染的MSCs组（Ⅰ组）12只、单纯移植MSCs组（Ⅱ组）12只和只注射无血清培养基的对照组（Ⅲ组）12只。移植术后4周,超声法检测心脏功能,并同术前比较;荧光显微镜下观察MSCs分布情况;免疫组化法检测梗死区微血管密度。结果 移植4周后Ⅰ 组的MSCs存活率大于其他2组;Ⅰ组的左室射血分数、E／A比值和梗死区微血管密度与其他2组比较差异有统计学意义（P〈0．05）。结论 VEGF165基因和MSCs联合策略是治疗心肌梗死的有效方法,所产生的协同治疗效果大于单纯的MSCs移植。     

Administration of bone marrow cells into surgically induced fibrocollagenous tunnels induces angiogenesis in ischemic rat hindlimb model

We established a comparative model of angiogenic induction in previously formed fibrocollagenous tunnels in rat inner thigh muscles. A unilateral hindlimb chronic ischemia model was performed in male Sprague-Dawley rats. A device was then inserted in the central portion of the inner thigh muscles. Vascularity in the ischemic limb was determined by means of an angiographic score, capillary/fiber ratio, and endothelial proliferation by histochemistry and immunohistochemistry. Autologous transplant of bone marrow, vascular endothelial growth factor (VEGF), or collagen-polyvinylpyrrolidone plus heparin induced significant vascularization of the ischemic hindlimb when compared to saline solution. However, the bone marrow group presented a higher angiographic score than the other two. No differences among groups were observed in capillary/fiber ratio or proliferation, except for the VEGF group, where capillary proliferating cells were significantly higher than in controls. Based on these results, bone marrow-derived progenitor cells may constitute a safe and viable alternative for the induction of therapeutic angiogenesis.     

Intraventricular infusion of vascular endothelial growth factor promotes cerebral angiogenesis with minimal brain edema

OBJECTIVE: Therapeutic cerebral angiogenesis, i.e., using angiogenic factors to enhance collateral vessel formation within the central nervous system, is a potential method for cerebral revascularization. Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen that also increases capillary permeability, particularly in ischemic tissue. The purpose of this study was to assess the angiogenic and capillary permeability effects of chronic intraventricular infusion of exogenous VEGF in nonischemic brain tissue, because many patients with impaired cerebrovascular reserve do not exhibit chronic cerebral ischemia. METHODS: Recombinant human VEGF(165) was infused into the right lateral ventricle of rats at a rate of 1 microl/h for 7 days, at concentrations of 1 to 25 microg/ml, with osmotic minipumps. Control animals received vehicle only. Vessels were identified in laminin immunohistochemical analyses. Capillary permeability and brain edema were assessed with Evans blue extravasation, [(3)H]inulin permeability, and brain water content measurements. RESULTS: Vessel density was dose-dependently increased by VEGF(165) infusions, with significant increases occurring in animals treated with 5 or 25 microg/ml, compared with control animals (P h 0.01). Significant enlargement of the lateral ventricles was observed for the highest-dose group but not for animals treated with other doses. Capillary permeability was assessed in animals treated with a dose of 5 microg/ml. An increase in capillary permeability in the diencephalon was identified with Evans blue extravasation and [(3)H]inulin permeability assessments; however, the brain water content was not significantly increased. CONCLUSION: Chronic intraventricular infusions of VEGF(165) increased vascular density in a dose-dependent manner. There seems to be a therapeutic window, because infusion of VEGF(165) at a concentration of 5 microg/ml resulted in a significant increase in vessel density with minimal associated brain edema and no ventriculomegaly.