微泡激励的超声空化对兔前列腺的损伤作用

目的 探讨微泡激励的超声空化效应对正常兔前列腺组织的损伤作用.方法 30只新西兰雄性兔随机分为超声微泡组、单纯超声组和微泡假照组进行实验.超声微泡组经静脉推注微泡0.1 ml/kg,超声辐照10 min;单纯超声组超声辐照10 min;微泡假照组仅静脉推注微泡0.1 ml/kg.各组循环注射伊文思蓝(EB)溶液示踪.随后视觉观察和定量分析前列腺EB漏出量,并行光镜检查.结果 超声微泡组的前列腺EB漏出量明显大于微泡假照组、单纯超声组(P＜0.01);光镜观察超声微泡组前列腺微血管破裂,组织间隙大量出血,血肿形成.结论 微泡激励的超声空化对兔前列腺有明显的机械损伤作用.     

诊断超声介导微泡提高肝纤维化通透性及促进基因传递

目的 探讨诊断超声联合微泡对肝纤维化组织通透性的影响及其介导基因转染肝纤维化大鼠的有效性。方法 采用二甲基亚硝胺(DMN)法建立大鼠肝纤维化模型,80只大鼠在建模第4周末随机分为:模型对照组、单纯微泡组、单纯超声组和诊断超声联合微泡组。分别进行肝纤维化微血管通透性实验和基因转染实验,采用激光共聚焦显微镜观察伊文思蓝(EB)在肝纤维化组织内分布情况,同时定量检测肝纤维化组织内EB的含量,评估不同分组微血管通透性。荧光显微镜下观察含增强型绿色荧光蛋白报告基因的质粒转染大鼠肝纤维化模型的基因表达情况。结果 激光共聚焦显微镜显示诊断超声联合微泡组纤维化肝实质内可见明显的EB红色荧光。诊断超声联合微泡组纤维化肝组织中EB含量明显高于其他3组(P<0.05)。荧光显微镜下观察,相比其余3组,诊断超声联合微泡组增强型绿色荧光蛋白最多,基因转染效率最高。结论 诊断超声联合微泡在提高纤维化肝脏微血管通透性的同时可促进基因传递。     

超声微泡造影剂对心肌组织毛细血管通透性的影响实验研究

目的　研究超声破坏微泡造影剂对靶区内心肌组织毛细血管通透性的影响 ;探讨超声微泡造影剂增强基因转染的机制。方法　 2 4只健康雄性 Wistar大鼠 ,取 15只分为 3组 ,第 1组经静脉输入含有伊文思蓝 (Evans blue,EB)的微泡造影剂 ,并采用超声波在鼠胸壁辐照约 6 min破坏心肌组织内造影剂 ;第 2组采用超声照射 ,同时经静脉单纯输入 EB溶液 ;第 3组单纯经静脉输入 EB作为对照。照射完毕 2 h后放血处死大鼠 ,使用标准曲线和分光光度法测量各组大鼠心肌组织中 EB含量 (作为反映血管通透性的指标 )。另 9只大鼠随机分为 3组 ,每组 3只。第 1组经静脉输入微泡造影剂 ,并采用超声波在鼠胸壁辐照约 6 min破坏心肌组织内造影剂 ;第 2组单纯采用超声照射 ;第 3组不加任何处理 ,作为对照。照射完毕即刻处死大鼠 ,取心肌组织进行电镜观察毛细血管的改变。结果　采用超声照射 ,并经静脉输入微泡造影剂的大鼠 ,心肌组织中 EB含量为 (75 .33± 16 .80 )μg/ g,比单纯超声照射[(32 .2 1± 9.5 3)μg/ g]及心肌正常组织 EB含量 [(37.16± 7.98)μg/ g]明显增加 (P<0 .0 5 )。电镜结果显示超声破坏微泡后 ,能使毛细血管破裂 ,红细胞溢出于毛细血管外。结论　超声波触发破坏微泡造影剂后能使心肌组织毛细血管通透性增加 ,可     

诊断超声联合微泡对大鼠移植瘤血管通透性影响的实验研究

目的 研究诊断超声联合微泡对大鼠移植瘤血管通透性的影响.方法 将48只已建立Walker-256皮下移植瘤模型的SD大鼠随机分为4组:空白对照(A)组、单纯微泡剂(B)组、单纯超声(C)组和超声联合微泡(D)组.以伊文思蓝(Evens blue,EB)为指示剂,选择频率为1.75 MHz、机械指数为1.4的诊断超声持续照射肿瘤区5 min,经大鼠尾静脉注射/不注射微泡造影剂.使用分光光度计和标准曲线法测量大鼠肿瘤组织内EB含量变化,激光共聚焦显微镜观察EB外溢情况.结果 超声联合微泡可增加肿瘤组织微血管通透性.D组瘤内EB含量较A、B、C组明显增加(P＜0.05),血管周围EB外溢明显.A、B、C组EB含量比较差异无统计学意义(P＞0.05),血管周围均未见明显EB渗出.结论 在一定条件下,诊断超声联合微泡可明显提高辐照肿瘤组织血管通透性,这对临床肿瘤化疗患者具有潜在的应用意义.     

低频超声联合微泡造影剂诱导人前列腺癌PC3细胞及DU145细胞自噬的实验研究

目的：研究低频超声联合微泡造影剂对人雄激素非依赖性前列腺癌PC3细胞及DU145细胞自噬的影响。方法2种细胞均各分为空白对照组、单纯微泡组、单纯低频超声组和低频超声联合微泡组4组,每组设3个复孔。单纯微泡组加200μl的微泡造影剂混匀,不进行超声辐照;单纯低频超声组以声功率80 mW的20 kHz低频超声,不加造影剂,连续波辐照60 s;低频超声联合微泡组加200μl的微泡造影剂,混匀后以声功率80 mW的20 kHz低频超声,连续波辐照60 s;空白对照组不进行任何处理,辐照后的细胞继续培养24 h,吖啶橙染色荧光显微镜与透射电镜观察细胞自噬泡。结果经吖啶橙染色后,荧光显微镜下,2种细胞低频超声联合微泡组的胞质内可见大量红色荧光的酸性囊泡,明显多于其余3组;单纯低频超声组红色荧光的酸性囊泡量亦多于空白对照组,空白对照组与单纯微泡组红色荧光的酸性囊泡量差异不大。透射电镜下,2种细胞的低频超声联合微泡组细胞细胞核基本正常,但胞质内出现大量由双层膜包裹的自噬泡或自噬体,而空白对照组内的细胞形态基本正常,胞质内未见到明显的自噬泡的形成。结论低频超声联合微泡造影剂能明显诱导人雄激素非依赖型前列腺癌PC3细胞及DU145细胞的自噬。     

不同参数对低频诊断超声联合微泡下肝血管通透性影响的研究

目的 探讨不同低频诊断级超声参数联合微泡对正常肝血管通透性的影响,优化超声参数以实现肝血管高通透性,减少对正常肝细胞的影响.方法 使用低频诊断超声辐照造影剂显影的肝脏,按实验分组设置不同水平的辐照时间、机械指数( mechanical index,MI)、微泡剂量,经大鼠尾静脉注入50 mg/kg伊文氏蓝(Evens blue,EB).观察辐照后肝组织中EB渗出情况,定量检测肝组织中EB含量的改变以探索不同参数条件下肝血管通透性的变化,同时对肝脏进行组织学检查观察细胞损伤情况.结果 随着辐照时间的延长、MI的升高、微泡浓度的增加,肝组织中EB含量均有明显增加;HE染色检测肝细胞无明显损伤.结论 低频诊断超声联合微泡可增加肝脏血管通透性,辐照时间、机械指数、微泡浓度是其重要影响因素.     

超声辐照微泡对大鼠前列腺增生组织通透性的影响

目的探讨大鼠前列腺增生组织通透性的改变及超声微泡对通透性的影响。方法24只SD大鼠随机分为正常组、前列腺增生组、超声辐照微泡治疗前列腺列增生组(UM BPH组)。UM BPH组注射白蛋白微泡并对前列腺局部用UGT1025型超声基因转染仪辐照。各组电镜观察其前列腺血管及细胞膜变化、计算前列腺指数(prostaticindex,PI),并测定前列腺组织中台盼蓝含量。结果电镜观察可见UM BPH组微血管基底膜变薄,部分血管周围有红细胞漏出,前列腺上皮细胞结构疏松,线粒体内可见空化;BPH组前列腺指数为2.88±0.03,UM BPH组为2.88±0.02,较正常组1.57±0.04均明显增加(P<0.05);前列腺组织中台盼蓝的含量:UM BPH组为(8.54±0.23)×10-9g/L,较BPH组[(2.54±0.11)×10-9g/L]及NP组[(4.20±0.22)×10-9g/L]明显增加(P<0.05)。结论在大鼠BPH时,组织通透性有所下降,超声辐照白蛋白微泡可以增加前列腺组织的通透性。     

诊断超声联合微泡增强骨髓间充质干细胞归巢兔缺血心肌的研究

目的 探讨诊断超声联合微泡对经静脉移植兔骨髓间充质干细胞(BM-MSCs)归巢缺血心肌的靶向能力.方法 采用密度梯度离心与贴壁法培养兔BM-MSCs,用DAPI标记细胞.完全结扎冠状动脉左前降支(LAD)法建立兔心肌梗死模型.将分离培养的BM-MSCs经静脉途径移植人心肌梗死兔体内.根据移植时是否介入超声与微泡分成单纯静脉移植MSCs组与超声辐照+微泡+MSCs组,移植后48 h荧光显微镜下观察缺血梗死区域及其周围的荧光标记阳性细胞数目,并对两组进行计数和统计学分析.HE染色观察局部缺血心肌的病理学改变.透射电镜观察心肌血管及内皮细胞情况.结果 荧光显微镜下显示超声辐照+微泡+细胞组较单纯静脉细胞移植组阳性细胞分布更为密集.单纯静脉细胞移植组与超声辐照+微泡+细胞组阳性细胞个数分别为(146.8±18.78)个和(213.2±26.5)个,差异有统计学意义(P<0.01).HE染色结果显示超声+微泡+细胞组缺血心肌及周围区心肌组织间隙可见红细胞成分漏出,而单纯细胞组无此发现.透射电镜显示在超声联合微泡的介导下,心肌血管内皮细胞间隔增大,血浆成分渗出.结论 超声联合微泡经静脉移植MSCs后能增加MSCs在缺血及周围区心肌的聚集与归巢,增强其靶向性.     

超声破坏微泡对心肌细胞膜通透性影响的研究

目的探讨超声破坏微泡造影剂对心肌细胞膜通透性的影响。方法将15只昆明小白鼠随机分为3组,一组采用尾静脉输入白蛋白微泡造影剂,胸壁用频率为1MHz,强度为1.5W/cm2的超声进行辐照1min;一组单纯采用同等超声辐照相同时间;一组作为对照。作用后即刻取心肌组织用硝酸镧(La)示踪法做透射电镜观察心肌细胞膜通透性的变化。结果采用超声破坏微泡后可见部分镧颗粒分布于心肌细胞胞浆内,部分分布于线粒体外、细胞核外;单纯超声作用组心肌细胞胞浆中可有少量的镧颗粒分布,而大部分镧颗粒分布于细胞膜外;而对照组镧颗粒分布于心肌细胞膜外。结论超声破坏微泡可提高心肌细胞膜通透性,可能是超声微泡造影剂增强组织中基因转染的机制之一。     

低频超声联合SonoVue微泡促进裸鼠基因体内转染的可行性

目的 探讨利用低频超声联合SonoVue微泡造影剂促进基因转染人前列腺癌裸鼠皮下移植瘤的可行性。方法 采用人前列腺癌PC-3细胞建立裸鼠皮下移植瘤模型,成瘤后随机分为空白组（仅注射生理盐水）、单纯质粒组（注射生理盐水和质粒混悬液）、低频超声组（注射生理盐水和质粒混悬液后超声辐照）和低频超声+微泡组（注射超声微泡造影剂和质粒混悬液后超声辐照）。超声辐照条件：功率3 W/cm²,占空比5：5,频率80 kHz,辐照10 min。基因转染3天后在激光共聚焦显微镜下观察各组绿色荧光蛋白表达情况,并分析其平均荧光强度;同时行常规病理检查,观察低频超声联合微泡辐照对组织的损伤程度。结果 4组中,仅低频超声+微泡组可见明显绿色荧光,与其他各组差异均有统计学意义（P均<0.05）;空白组、单纯质粒组和低频超声组均未见明显绿色荧光。各组均未见明显组织损伤。结论 SonoVue微泡造影剂在低频超声辐照下能够安全有效地促进pEGFP基因转染进入人前列腺癌裸鼠皮下移植瘤组织内。     

超声声致孔隙效应致兔前列腺损伤的初步研究

目的 探讨超声联合造影剂微泡的声致空隙效应开放血-前列腺屏障时对前列腺的损伤作用.方法 15只健康8月龄性成熟新西兰兔,随机分为单纯微泡(MB)组、单纯超声(US)组、超声联合微泡(US+MB)组,超声波直接辐射前列腺,光镜、电镜、细胞凋亡指数(AI)观察前列腺组织损伤的情况.结果 MB组与US组前列腺组织在光镜下均未见明显异常,胞浆、细胞核染色均匀,腺体上皮组织排列整齐、完整,腺管腔内未见明显改变;US+MB组腺体上皮组织细胞排列紊乱,腺管腔内可见大量嗜伊红染的液体.透射电镜下MB组与US组前列腺组织微血管内皮细胞排列整齐,细胞间紧密连接形态正常;US+MB组可见线粒体肿胀,微血管内皮细胞间隙增宽,细胞间桥连接断裂,红细胞漏出等改变.US+MB组AI明显高于MB组与US组(P<0.01),而US组则明显高于MB组(P<0.01).结论 利用超声联合造影剂微泡的声致孔隙效应开放血-前列腺屏障的同时可造成前列腺组织损伤.

Abstract：

Objective To explore the damage effect of sonoporation on the prostate of rabbit,while opening up the blood-prostate barrier by microbubble mediated sonoporation.Methods Fifteen male New Zealand white rabbits were randomly divided into 3 groups:ultrasound (US) group,microbubble (MB) group,ultrasound and microbubble (US+MB) group.Ultrasound was insonated directly on the prostate.Optical microscope,electron microscope and apoptosis index (AI) with TUNEL method were applied to trace the changes of the prostate of rabbit under different conditions.Results There was no significant change in prostatic tissues of group US and MB under the optical microscope.Cytoplasm and nucleoli were stained equally,cells of glandular epithelium were intact and formed orderly.Glandular cavities in these two groups were change very slightly.Glandular epithelium cells of Group US+MB were organized optical under the optical microscope,and there was a mass of eosinophilic liquid in the glandular cavities.Vascular endothelial cell were intact and formed orderly and swollen mitochondria were observed under the electron microscope in MB group and US group.Swollen mitochondria,tight junctions among gland cells were opened,and infiltrated erythrocyte could be found under the eletron microscope in US+MB.AI of group US+MB was markedly higher than that of group US and group MB (P<0.01),and AI of group US was higher than that of group MB (P<0.01).Conclusions Microbubble mediated sonoporation causes damage in the prostate tissue of rabbit,while opening up the blood-prostate barrier with an increased permeability of the prostate.     

Ultrasound-Mediated Microbubble Destruction Increases Renal Interstitial Capillary Permeability in Early Diabetic Nephropathy Rats

Diabetic nephropathy (DN) is defined as persistent proteinuria corresponding to a urinary albumin excretion rate >300 μg/mg in the absence of other non-diabetic renal diseases. The aim of this study was to determine if ultrasound (US)-mediated microbubble (MB) destruction could increase renal interstitial capillary permeability in early DN rats. Diabetes was induced with streptozotocin. DN rats presented with mild micro-albuminuria 30 d after onset of diabetes. DN rats (N = 120) were divided into four groups that received Evans blue (EB) followed by: (i) no treatment (control group); (ii) continuous ultrasonic irradiation for 5 min (frequency = 7.00 MHz, mechanical index = 0.9, peak rarefactional pressure = 2.38 MPa: US group); (iii) microbubble injection (0.05 mL/kg: MB group); and (iv) both ultrasound and microbubble injection (US + MB group). Another 8 DN rats were subjected to ultrasound and microbubbles and then injected with EB after 24 h (recovery group). EB content, EB extravasation and E-selectin mRNA and protein expression significantly increased, and interstitial capillary walls became discontinuous in the US + MB group. Neither hemorrhage nor necrosis was observed on renal histology. Urine samples were collected 24 h post-treatment. There was no hematuria, and the urinary albumin excretion rate did not increase after ultrasound-microbubble interaction detected by urinalysis. EB content returned to the control group level after 24 h, as assessed for the recovery group. In conclusion, ultrasound-mediated microbubble destruction locally increased renal interstitial capillary permeability in DN rats, and should be considered a therapy for enhancing drug and gene delivery to the kidney in the future.     

超声破坏微泡对大鼠皮肤创面愈合影响的实验研究

目的 探讨超声破坏微泡(声诺维)对大鼠皮肤创面愈合的影响.方法 96只SD大鼠建立皮肤创面模型后随机分成4组:超声破坏微泡组、单纯微泡组、单纯超声组和对照组.超声破坏微泡组经鼠尾静脉注射微泡造影剂0.5 ml,同时用声强度1 MHz、2.0 W/cm2超声辐照3 min单纯微泡组经鼠尾静脉注射微泡造影剂0.5 ml;单纯超声组经鼠尾静脉注射生理盐水0.5 ml,同样条件超声辐照3 min对照组经鼠尾静脉注射生理盐水0.5 ml.于创伤后第1、3、5、7、14、21 d利用HE染色和免疫组化方法 观察各组创面肉芽组织生长及血管内皮生长因子(VEGF)表达的情况.结果 HE染色:伤后第7 d,超声破坏微泡组肉芽组织明显比其他3组厚,新生毛细血管均垂直于创面生长,其他3组毛细血管排列紊乱.免疫组化观察VEGF表达变化:超声破坏微泡组在第3 d形成表达高峰,其他3组表达高峰在第5～7 d.结论 超声破坏微泡可以提高大鼠背部皮肤的创面愈合质量;新生肉芽组织成熟早,创面愈合加速.超声破坏微泡时产生的高温、高压及某些化学效应可以刺激内源性VEGF分泌,促进血管生成,从而促进创面愈合.

Abstract：

Objective To investigate the effect of microbubble(Sono Vue) destruction with ultrasound on wound healing in rats. Methods Total 96 SD rats were accepted one rounded whole-layer skin incision on back each other and randomly divided into four groups:microbubble destruction with ultrasound(US + MB),microbubble(MB), ultrasound(US) and control group. Rats in US + MB group were injected with 0.5 ml microbubble contrast agent via tail vein,and then ultrasound irradiated for 3 minutes immediately. MB group were injected with 0.5 ml microbubble contrast agent. US group were injected with 0.5 ml physiological saline,and then ultrasound irradiated for 3 minutes immediately under the same condition. Control group were injected with 0.5 ml physiological saline. Feed each rat in single cage. On day 1,3,5,7,14 and 21 after wound creation,the excised wound tissues were analyzed by histology and VEGF expression in wounds by immunohistochemistry. Results HE staining: On day 7, wounds of US + MB group displayed the most accumulation of granulation tissue and all new capillaries were perpendicular to the wound surface, but the new capillaries of other 3 groups were disordered. Immunohistochemical examination of VEGF expression:the peak expression appeared on day 3 in US + MB group, other 3 groups were on day 5 to day 7.Conclusions US + MB treatment could improve the quality of wound healing and granulation tissues were maturated earlier than MB, US treatment and control group, which could accelerate wound healing. High temperature,high pressure and some kind of chemistry effecs induced by microbubble destruction with ultrasound can stimulate the secretion of endogenous VEGF, which may be the mechanism of promoting angiogenesis and wound healing.     

超声击破微泡效应抑制兔VX_2肿瘤生长的实验研究

目的 探讨超声击破微泡效应抑制兔VX_2肿瘤生长的可行性.方法 21只移植有皮下VX_2肿瘤的新西兰大白兔,随机分为超声微泡治疗组、单纯超声组和假照组.经静脉注射脂质微泡的同时,脉冲式聚焦超声直接辐照肿瘤区域.单纯超声组和假照组分别用5 ml生理盐水和超声假照替代,每次10 min,每隔72 h治疗一次,采集各时间点肿瘤二维声像图和超声造影影像,测量肿瘤切面最大直径.结果 在30 d的实验周期内,微泡超声治疗组、单纯超声组和假照组的肿瘤平均直径分别从(1.1±0.1)cm、(1.2±0.1)cm、(1.2±0.1)cm生长至(2.1±0.5)cm、(3.0±0.9)cm、(3.4±0.7)cm,微泡超声治疗组肿瘤直径明显小于另两组.结论 超声击破微泡效应可阻断肿瘤微循环,有一定的抑制肿瘤生长作用.     

超声靶向破坏微泡介导骨髓间充质干细胞移植促进大鼠血管新生

目的 探讨超声破坏微泡介导骨髓间充质干细胞(MSCs)移植在大鼠缺血骨骼肌中的作用. 方法 24只Wister大鼠离断右侧股动脉7 d后分为4组:①超声+微泡组(US+MB);②干细胞静脉移植组(MSCs-iv);③超声+微泡+干细胞静脉移植组(US+MB+MSCs-iv);④对照组.处理后第7 d取局部缺血骨骼肌行免疫组化检测. 结果 US+MB+MSCs-iv组缺血骨骼肌可见较多移植的MSCs,其新生血管数量较其他组明显增加. 结论 超声靶向破坏微泡有可能成为一种增强MSCs移植和促进血管新生的新方法 .     

Investigation Into the Impact of Diagnostic Ultrasound with Microbubbles on the Capillary Permeability of Rat Hepatomas

Ultrasound-targeted microbubble destruction (UTMD) takes advantage of transiently increased capillary permeability to enhance the release of tumor-specific drugs from blood vessels into sonicated tumor tissues. However, the application of focused ultrasound is limited because of the lack of an appropriate image-monitoring system. In this study, hepatoma-bearing Sprague-Dawley rats were insonicated with low-frequency diagnostic ultrasound and injected with Evans Blue (EB) dye and microbubbles through their tail veins to test changes in capillary permeability. We studied how the mechanical index, sonication duration and the injected microbubble (MB) concentration affect the hepatoma vascular permeability by quantitatively evaluating the EB delivery efficiency. Confocal laser scanning microscopy was used to observe the deposition of red fluorescence–dyed EB in tumor tissues. In addition, P-selectin, a type of biochemical marker that reflects vascular endothelial cell activation, was identified using an immunoblotting analysis. The experimental results reveal that EB delivery efficiency in tumor tissues was greater in groups with the diagnostic ultrasound–mediated UTMD (8.40 ± 0.71 %ID/g) than in groups without UTMD (1.73 ± 0.19 %ID/g) and EB delivery efficiency could be affected by MI, sonication duration and MB dose. The immunoblotting analysis indicates that diagnostic ultrasound–induced UTMD results in the vascular endothelial cell activation to increase capillary permeability, justifying the high quantity of EB deposited in tumor tissues.     

低频诊断超声联合脂氟显微泡开放小鼠血脑屏障的可逆性研究

目的探讨低频诊断超声联合脂氟显微泡开放小鼠血脑屏障(blood-brain barrier,BBB)的可逆性及伊文思蓝(Evans blue,EB)透过率。方法健康C57BL/6小鼠48只,随机分为对照组6只和观察组42只。观察组再随机分为0、0.5、1、2、3、8、10 h亚组各6只,低频诊断超声辐照同时经尾静脉注射脂氟显微泡1 mL/kg,并分别于超声辐照0、0.5、1、2、3、8、10 h经尾静脉注射质量分数2%EB溶液50 mg/kg;对照组不进行低频诊断超声辐照,经尾静脉注射1 mL/kg生理盐水后注射质量分数2%EB溶液50 mg/kg。注射1 h后处死小鼠取脑组织,观察脑组织蓝染深度及面积,采用紫外分光光度法定量分析脑组织EB含量。结果对照组及观察组8、10 h亚组未见脑实质蓝染,观察组0、0.5、1、2、3 h亚组辐照区脑实质均有不同程度蓝染,且1 h时蓝染深度最深、面积最大;观察组0、0.5、1、2、3 h亚组脑组织EB含量[(45.43±2.89)、(50.94±1.25)、(79.79±1.12)、(51.55±1.20)、(31.60±1.77)μg/g]均高于对照组[(8.32±0.12)μg/g](P<0.05),8、10 h亚组脑组织EB含量[(8.34±0.09)、(8.31±0.07)μg/g]与对照组比较差异无统计学意义(P>0.05);观察组0、0.5、1 h亚组脑组织EB含量依次升高(P<0.05),1、2、3 h亚组脑组织EB含量依次降低(P<0.05)。结论低频诊断超声联合脂氟显微泡开放小鼠BBB呈动态可逆的过程,BBB在超声辐照后1 h开放程度最大,于超声辐射8 h恢复正常。     

Experimental research on therapeutic angiogenesis induced by hepatocyte growth factor directed by ultrasound-targeted microbubble destruction in rats.

OBJECTIVE: The purpose of this study was to explore the feasibility of therapeutic angiogenesis in myocardial infarction induced by hepatocyte growth factor (HGF) mediated by ultrasound-targeted microbubble destruction. METHODS: Forty Wistar rats were divided into 4 groups after the models of myocardial infarction were prepared: (1) HGF, ultrasound, and microbubbles (HGF+US/MB), (2) HGF and ultrasound, (3) HGF and microbubbles, and (4) surgery alone. Destruction of ultrasound-targeted microbubbles loaded with the HGF gene with an electrocardiographic trigger mode was performed in the HGF+US/MB group. All the rats were killed after being transfected for 14 days. Enhanced green fluorescent protein expression was examined in the myocardium, liver, and kidney in all groups by fluorescence microscopy; CD34 expression was detected by immunohistochemistry, and microvessel density (MVD) was counted in the high-power field on microscopy. Hepatocyte growth factor expression in the myocardium was detected by western blotting and an enzyme-linked immunosorbent assay. RESULTS: Enhanced green fluorescent protein expression was detected in the myocardium of the HGF+US/MB group, but a few areas of HGF expression were detected only in small vessels and the capillary endothelium, and no expression was found in the surgery-alone and HGF and microbubbles groups. The results of MVD counting by microscopy showed that the MVD in the myocardium of the HGF+US/MB group was the highest among all the groups. The results of western blotting and the enzyme-linked immunosorbent assay showed that the amount of HGF in the myocardium was highest in the HGF+US/MB group. CONCLUSIONS: Ultrasound-targeted microbubble destruction could deliver HGF into the infracted myocardium and produce an angiogenesis effect, which could provide a novel strategy for gene therapy of myocardial infarction.     

超声击破造影剂微泡阻断正常肝血流灌注的造影观察

目的 采用视觉评分方法分析微泡增强的超声空化阻断兔正常肝血流灌注后的恢复情况.方法 健康新西兰兔24只,分为超声微泡组、单纯超声组及假照组.超声微泡组经兔耳缘静脉推注脂质微泡联合超声辐照兔肝,单纯超声组以生理盐水代替微泡,假照组超声假照.各组治疗前及治疗后0 min、15 min、30 min、45 min、60 min和24 h对兔肝进行超声造影,视觉评分分析各时间点造影灌注峰值灰阶变化.结果 各组治疗前肝血流灌注比较,视觉评分差异无统计学意义(P>0.05);超声微泡组的治疗前造影血流灌注评分显著优于治疗后0 min、15 min两个时间点,差异有统计学意义(P<0.05);但与治疗后30 min、45 min、60 min、24 h视觉评分差异无统计学意义(P>0.05);而单纯超声组、假照组的治疗前后各时间点之间超声造影评分差异均无统计学意义(P>0.05).结论 微泡增强的超声空化具有显著的暂时性阻断兔肝实质血流作用.     

靶向和非靶向微泡联合尿激酶超声溶栓的电镜表现

目的 比较靶向和非靶向微泡联合尿激酶超声溶栓的电镜表现。方法 将精氨酸-甘氨酸-天冬氨酸-丝氨酸片段（RGDS）与尿激酶（UK）以及超声微泡（SonoVue）通过机械振荡法,制备成靶向微泡。于30只新西兰大白兔单侧股动脉制备在体混合性血栓,并分为单纯超声辐照组（US组）、超声辐照+非靶向微泡造影剂+UK组（US+M+UK组）、超声辐照+靶向微泡造影剂+UK组（US+RGDS+UK组）。通过超声及多普勒血流仪观察溶栓效果,然后对股动脉离体标本行HE染色,并观察电镜表现。结果 溶栓20 min后,与US组和US+M+UK组比较,US+RGDS+UK组血流量明显恢复（P均<0.05）,US组与US+M+UK组比较差异无统计学意义（P均>0.05）。US+M+UK组HE染色显示管腔内充满血栓,血小板梁呈颗粒状、不致密,扫描电镜示粗大束状的胶原纤维上疏松附着少量细小纤维蛋白丝,大部分断裂;透射电镜示血栓大部分溶解为空泡状,可见白细胞或血小板降解的碎片。US+RGDS+UK组HE染色显示血栓完全溶解;扫描电镜示血栓的纤维网状结构被破坏,纤维蛋白完全的溶解;透射电镜示血栓降解为高电子密度的颗粒。结论 血栓结构的空泡化、纤维蛋白网架结构完全崩解和纤维蛋白的完全溶解是靶向微泡和UK联合溶栓的主要电镜改变。