高强度聚焦超声辐照兔腹主动脉粥样硬化血管后的病理学变化

目的观察高强度聚焦超声(high intensity foeused ultrasound，HIFU)辐照后兔腹主动脉粥样硬化(athero—sclerosis，AS)血管的病理学变化。方法通过大体、光镜和透射电镜观察HIFU辐照后不同时间，兔腹主动脉AS血管的病理学变化。结果48只兔(AS组：n1=24，对照组：n2=24)，HIFU辐照后腹主动脉出现一定程度损伤，但无血管栓塞和破裂；辐照后5d、10d血管损伤呈逐渐恢复趋势，对照组恢复更好。结论HIFU辐照导致兔腹主动脉粥样硬化血管产生可逆性损伤。     

动脉粥样硬化经高强度聚焦超声作用后血管的结构和功能变化

目的 观察高强度聚焦超声（HIFU）在体辐照兔粥样硬化腹主动脉后血管的结构和功能变化，探讨HIFU对病理性血管的影响。方法建立粥样硬化兔动物模型，经HIFU辐照后，通过光镜、透射电镜、彩色多普勒血流显像（CDFI）和数字减影血管造影（DSA）测定血管组织形态学和血流动力学变化。结果 经HIFU辐照后，兔动物模型粥样硬化腹主动脉出现一定程度损伤，但无血管栓塞和破裂，实验组和对照组血管损伤程度差异无统计学意义；CDFI系列血流参数均下降（P〈0．05）；DSA显示动脉血流阻滞，充盈延迟，无血管栓塞和破裂。辐照后5d及10d观察损伤血管呈逐渐恢复趋势，动脉血流参数呈恢复趋势，对照组恢复更好。结论 HIFU辐照兔粥样硬化腹主动脉将导致可逆性损伤和血流动力学改变。     

高强度聚焦超声联合超声造影剂消融兔肝后细胞凋亡和增殖细胞核抗原表达的研究

目的：探讨高强度聚焦超声（HIFU）联合超声造影剂消融技术对兔肝组织细胞凋亡和增殖细胞核抗原（PCNA）表达的影响。方法：40只新西兰纯种大白兔分为A、B组，A组兔肝接受HIFU辐照前，经耳缘静脉注射生理盐水；B组接受辐照前注射超声造影剂SonoVue。A、B组均在辐照后5min、1d、3d、7d、14d分别处死4只动物，将肝取出体外观察，取靶区及周围组织（距靶区边缘5mm以内），HE染色观察组织病理学改变，原位末端标记法（TUNEL）检测细胞凋亡，免疫组织化学方法检测细胞PCNA表达。结果：HIFU辐照后3d、7d、14dB组辐照区域边缘纤维包裹带宽度大于A组（P〈0．05）。HIFU辐照后1d、3d、7d、14dB组靶区周围组织（距靶区边缘5mm以内）凋亡细胞数和PCNA阳性细胞指数较A组增高（P〈0．05）。结论：与单纯HIFU相比，HIFU联合超声造影剂促进了消融后坏死区域机化包裹，增加了HIFU靶区周围细胞凋亡，提高了细胞增殖能力，为进一步增强HIFU消融作用提供了实验数据。     

HIFU消融后兔肝细胞凋亡与增殖的研究

目的探讨高强度聚焦超声（HIFU）消融兔肝组织后细胞凋亡和增殖细胞核抗原（PCNA）表达的变化。 方法24只新西兰大白兔肝组织接受HIFU辐照，辐照后5min，1、3、6、9、14d分别处死4只动物，取出肝脏，HE染色观察组织病理改变，透射电镜观察细胞超微结构变化，原位末端标记法（TUNEL）检测细胞凋亡，免疫组织化学方法检测PCNA表达。 结果HE染色显示HIFu辐照后靶区边缘形成纤维包裹带，随时间逐渐加宽（P〈0．05）。电镜发现辐照后靶区周围组织有凋亡小体生成，TUNEL检测凋亡指数于辐照后3d最高。辐照后靶区周围组织出现PCNA阳性细胞，PCNA阳性细胞指数于辐照后6d达到峰值。 结论HIFU辐照后凝固坏死区域逐渐机化包裹，靶区周围组织出现凋亡细胞和PCNA阳性细胞，数量随时间变化，为进一步控制和增强HIFu消融作用提供实验基础。     

高强度聚焦超声阻断毫米级兔股动脉血流的实验研究

目的 探讨高强度聚焦超声(HIFU)通过体外照射能否导致毫米级兔股动脉内血栓形成,血流被较长期阻断.方法 HIFU工作频率0.4 MHz,焦距110 mm,焦点平均强度2.0 kW/cm2,将60只兔随机分成三组,用HE染色观察HIFU照射后即日、3 d、7 d兔股动脉组织学变化,DSA观察HIFU照射后兔股动脉血流变化.结果 HIFU照射后各实验组所有兔均可见股动脉内血栓形成,数字减影血管造影检查显示兔股动脉内血流中断.结论 HIFU照射能使毫米级的兔股动脉在较长的时间内(7 d)血栓形成,血流中断.     

三种化学方法处理异体小血管移植的实验

目的：比较3种化学方法处理异体小血管移植后血管通畅状态以及病理变化。方法：实验于2005-03／09在中山大学医学院动物实验中心完成。①采用随机数字表法将64只新西兰白兔随机分成4组，自体组，甘油组，乙醇组和戊二醛组，每组16只。（2）自体组以自体股动脉移植作为对照；甘油组，乙醇组和戊二醛组，分别将980g／L甘油、体积分数为0.95的乙醇、5g／L戊二醛保存的异体兔股动脉，移植桥接于兔股动脉。③各组兔分别于术后7，14，28，56d观察血管通畅情况，同时取移植体标本观察其病理变化。结果：①各组兔股动脉通畅率：甘油组和自体血管移植8周后通畅率均为100％；乙醇组8周后通畅率为69％，戊二醛组通畅率为6％；甘油通畅率明显高于乙醇组和戊二醛组（P〈0.01）。②各组兔移植前血管组织学变化：甘油浸泡后的血管复苏后外观颜色、质地与正常血管相同，血管弹性良好。乙醇浸泡后的血管其外观颜色、质地与正常血管相同，但血管弹性稍差。戊二醛组血管颜色略变黄，血管弹性欠佳。③异体血管移植后组织学检查结果：甘油组术后2周开始，血管内壁内皮细胞开始增生，至8周时明显。弹力纤维8周时仍可见排列尚整齐。乙醇组术后移植血管内壁内皮细胞变化基本同甘油组。中层平滑肌细胞以及弹力纤维移植后2周开始出现变性或结构紊乱。戊二醛组从2周开始，管腔封闭，弹力纤维结构紊乱。4墙周管腔内充满较多的成纤维细胞。结论：3种化学方法中，甘油处理的异体小动脉移植在8周内与自体血管移植通畅率无明显差别，在8周内可保证通畅。     

HIFU辐照兔腹主动脉粥样硬化血管后血流动力学的变化

目的　观察HIFU辐照兔腹主动脉粥样硬化 (atherosclerosis ,AS)血管后血流动力学的变化 ,确定HIFU的安全性。方法　以CDFI和DSA检测HIFU辐照兔AS及对照组腹主动脉后不同时间血管血流的变化 ,测定血流动力学指标。结果　 48只兔 (AS组 :n1=2 4,对照组 :n2 =2 4) ,HIFU辐照后CDFI示动脉峰值血流速度、平均血流速度、阻力指数、搏动指数、血流量及血流量下降百分比均下降 (P <0 .0 5 )。DSA示动脉血流阻滞、充盈延迟 ,无血管栓塞和破裂 ;5d及 10d观察动脉血流呈向正常恢复趋势 ,对照组恢复更好。结论　HIFU辐照兔腹主动脉AS血管将导致可逆的血流动力学改变 ,HIFU辐照兔腹主动脉粥样硬化血管具有相对安全性     

超声造影剂增强HIFU消融兔肝组织的电镜观察

目的探讨高强度聚焦超声(HIFU)联合超声造影剂消融后兔肝细胞超微结构变化。方法30只动物分为两组,A组单纯HIFU辐照,B组辐照前注射超声造影剂。辐照后1h、6d、14d电镜观察。结果辐照后两组靶区细胞坏死,辐照后早期A组靶区可见细胞轮廓。辐照后6d两组靶区细胞崩解,边缘见纤维包裹带,靶区周围细胞浊肿,B组较A组严重,B组靶区周围凋亡小体多于A组。结论超声造影剂增强HIFU消融作用,进一步破坏组织超微结构。     

高强度聚焦超声对人乳腺癌细胞及其滋养血管损伤的病理观察

目的：观察高强度聚焦超声（HIFU）对乳腺癌及营养血管的破坏作用。方法：对23例乳腺癌患者行HIFU治疗，每例作2张连续的切片，分别进行HE染色、弹力纤维维多利亚蓝和丽春红S组织化学染色，观察HIFU对乳腺癌营养血管破坏的情况。结果：靶区乳癌组织及小于2mm营养血管完全破坏，血管弹力纤维出现崩解、分布不均、断裂及分层等现象，而邻近组织大血管未受损伤。结论：HIFU能完全破坏乳癌细胞及其营养血管，从而有可能抑制肿瘤的增生和转移，加强HIFU治疗肿瘤的效果。HIFU治疗乳癌患者安全、有效、可行。     

三种化学方法处理异体小血管移植的实验

目的:比较3种化学方法处理异体小血管移植后血管通畅状态以及病理变化。方法:实验于2005-03/09在中山大学医学院动物实验中心完成。①采用随机数字表法将64只新西兰白兔随机分成4组,自体组,甘油组,乙醇组和戊二醛组,每组16只。②自体组以自体股动脉移植作为对照;甘油组,乙醇组和戊二醛组,分别将980g/L甘油、体积分数为0.95的乙醇、5g/L戊二醛保存的异体兔股动脉,移植桥接于兔股动脉。③各组兔分别于术后7,14,28,56d观察血管通畅情况,同时取移植体标本观察其病理变化。结果:①各组兔股动脉通畅率:甘油组和自体血管移植8周后通畅率均为100%;乙醇组8周后通畅率为69%,戊二醛组通畅率为6%;甘油通畅率明显高于乙醇组和戊二醛组(P<0.01)。②各组兔移植前血管组织学变化:甘油浸泡后的血管复苏后外观颜色、质地与正常血管相同,血管弹性良好。乙醇浸泡后的血管其外观颜色、质地与正常血管相同,但血管弹性稍差。戊二醛组血管颜色略变黄,血管弹性欠佳。③异体血管移植后组织学检查结果:甘油组术后2周开始,血管内壁内皮细胞开始增生,至8周时明显。弹力纤维8周时仍可见排列尚整齐。乙醇组术后移植血管内壁内皮细胞变化基本同甘油组。中层平滑肌细胞以及弹力纤维移植后2周开始出现变性或结构紊乱。戊二醛组从2周开始,管腔封闭,弹力纤维结构紊乱。4～8周管腔内充满较多的成纤维细胞。结论:3种化学方法中,甘油处理的异体小动脉移植在8周内与自体血管移植通畅率无明显差别,在8周内可保证通畅。     

Application of high-intensity focused ultrasound for umbilical artery occlusion in a rabbit model.

OBJECTIVES: To investigate the application of high-intensity focused ultrasound (HIFU) for fetal umbilical artery blood flow occlusion in a rabbit model. METHODS: A prototype HIFU transducer in combination with an imaging probe with Doppler capability was constructed. Using this transducer, HIFU was applied at 1.4, 2.75 or 5.5 kW/cm(2) through the maternal abdominal skin to the fetal intra-abdominal umbilical arteries of four time-mated Japanese White rabbits (11 fetuses) on gestational day 25. Courses of 5-s HIFU exposure were performed until cessation of umbilical blood flow and cardiac arrest were confirmed by Doppler ultrasonography. Fetal necropsy was performed and exposed lesions were assessed by microscopic histological analysis. RESULTS: The mean diameter of the fetal umbilical artery was 0.6 +/- 0.2 mm and the mean peak systolic velocity of arterial blood flow was 44.7 +/- 18.5 cm/s. When HIFU was applied at 5.5 kW/cm(2), blood flow was completely occluded within 15 courses. HIFU exposure brought about vacuolar degeneration and destruction of elastic fibers in the tunica media of the artery. CONCLUSIONS: HIFU can be used to occlude umbilical artery blood flow in fetal rabbits.     

Functional and histological changes in rat femoral arteries by HIFU exposure

This study was an investigation of arterial contractility in response to high-intensity focused ultrasound (HIFU) and of histologic changes to the artery with various intensities of HIFU. We constructed a prototype HIFU transducer in combination with an imaging probe that provides color Doppler imaging and Doppler velocimetry. HIFU was applied through the skin to deep femoral arteries in left thighs of Sprague-Dawley rats; color images of the blood flow were used to aim the HIFU beam. Peak intensities used were 530, 1080, 2750 and 4300 W/cm2. The duration of each HIFU exposure was 5 s. HIFU was applied to five focal spots of each leg. These focal spots were aligned with a spacing of 1.0 mm so as to form a line across the artery. Blood flow occlusion was accomplished by HIFU at an intensity of 4300 W/cm2, but the flow continued with the lower intensities. Peak systolic velocities (PSVs) of blood flow as measured by Doppler velocimetry increased in the arteries to which HIFU had been applied at 1080 and 2750 W/cm2. The increase corresponded with HIFU intensity. Exposure to HIFU at 530 W/cm2 did not change the blood flow velocity. Histologic studies have demonstrated that exposure to HIFU at 2750 and 4300 W/cm2 leads to vacuolar degeneration and destruction of elastic fibers of the tunica media of the artery. Exposure at 1080 W/cm2 led to increased PSV, but did not induce histologic changes in the vessel wall. In conclusion, the response of the artery to HIFU varied with intensity. Vascular contraction without tissue degeneration occurred at low intensity; with increasing intensity, the tissue degeneration detectable in histology reduced the vascular diameter and, finally, at high intensity, the blood flow was occluded. Although these phenomena appeared to be mainly due to thermal effects, mechanical effects might have some role, particularly on vascular contraction.     

Pulsatile flow phantom for ultrasound image-guided HIFU treatment of vascular injuries

A pulsatile flow phantom was developed for studies of ultrasound image-guided high intensity focused ultrasound (HIFU) application in transcutaneous hemostasis of injured blood vessels. The flow phantom consisted of a pulsatile pump system with instrumented excised porcine carotid artery, which was imbedded in a transparent agarose gel to model structural configuration of in vivo tissues. Heparinized porcine blood was circulated through the phantom. The artery was injured using an 18-gauge needle to model a penetrating injury in human peripheral vasculature. A HIFU transducer with the diameter of 7 cm, focal length of 6.3 cm and frequency of 3.4 MHz was used to seal the puncture. Ultrasound imaging was used to localize and target the puncture site and to monitor the HIFU treatment. Triphasic blood flows present in the human arteries were reproduced, with flow rates of 50 to 500 mL/min, pulse rates of 62 to 138 beats/min and peak pressures of 100 to 250 mm Hg. The penetrating injury of an artery was mimicked successfully in the flow phantom setting and was easily visualized both optically through the transparent gel and with power Doppler ultrasound imaging. Hemostasis was achieved in 55 +/- 31 s (n = 9) of HIFU application. Histologic observations showed that a HIFU-sealed puncture was filled with clotted blood and covered with a fibrin cap. The pulsatile flow phantom provides a controlled and repeatable environment for studies of transcutaneous image-guided HIFU application in hemostasis of a variety of blood vessel injuries.     

高强度聚焦超声致兔胃壁损伤的实验研究

目的 探讨模拟临床照射剂量条件下高强度聚焦超声(HIFU)对兔胃壁的破坏程度，以了解HIFU技术的安全性。方法 HIFU体外照射20只新西兰大白兔胃壁，观察照射过程中超声影像改变、局部损伤区的组织病理变化以及术后生存情况。结果 监控超声图像可较敏感的反映出HIFU照射组织损伤区的变化，术后病理观察可以证实兔胃壁发生急性坏死。结论 临床治疗剂量的HIFU照射足以使胃壁发生急性坏死，致胃穿孔发生导致死亡。     

Sequential changes in rat femoral artery blood flow and tissue degeneration after exposure to high-intensity focused ultrasound

Purpose  The purpose of this study was to investigate the sequential changes in rat artery blood flow and tissue degeneration after exposure to high-intensity focused ultrasound (HIFU) in vivo. Methods  HIFU was applied through the skin to the femoral artery of Sprague-Dawley rats. The peak intensities used were 530, 1080, 2750, and 4300 W/cm². After exposure, we measured the peak systolic velocity (PSV) in the artery every 1 min until the velocity stabilized. The vessel was resected and examined histologically 7 days after exposure. Results  PSV was not significantly affected by HIFU exposure at 530 W/cm². PSV increased immediately after HIFU exposure at intensities of 1080 and 2750 W/cm². PSV after HIFU exposure at 1080 W/cm² fell to the control level within minutes; however, PSV increased immediately after HIFU exposure at 2750 W/cm² and then decreased slowly but remained at a higher level than the control for 15 min. On HIFU exposure at 4300 W/cm², the target artery was completely occluded. Histological studies 7 days after HIFU exposure demonstrated that exposure at 530 and 1080 W/cm² induced vacuolar degeneration in the tunica media of the femoral artery in rats; exposure to HIFU at 2750 and 4300 W/cm² resulted in strong necrotic degeneration in the tunica media. These histological changes were more marked than those found immediately after HIFU exposure. Organized thrombus formation was observed only for HIFU exposure at 4300 W/cm². Conclusion  Sequential changes in arterial blood flow after HIFU exposure vary with the intensity, and the histological changes in arterial tissue progress over time. These phenomena should be considered when HIFU is clinically applied to achieve arterial occlusion.     

超声引导下兔急性肺动脉高压模型的建立及其意义

目的探讨超声引导下肺栓塞所致兔急性肺动脉高压动物模型的建立及其意义。方法模拟人体血栓栓子脱落造成急性肺动脉栓塞的过程，采用超声引导下股静脉穿刺右心导管肺动脉放置以及压力检测，同步经对侧股静脉穿刺推注自体血栓，建立兔急性肺动脉高压模型。结果实验用新西兰兔30只，成功建立急性肺动脉高压模型25只，另5只发生急性右心衰竭死亡，模型成功率83％；超声检测栓塞后即刻出现三尖瓣反流，估测肺动脉收缩压与右心导管测定值高度相关（r=0．765，P=0．002）。结论采用超声引导下建立肺动脉血栓栓塞所致兔急性肺动脉高压模型的方法简单易行，成功率高，为急性肺动脉高压的基础和临床研究提供了一个良好的动物模型平台。     

碱性成纤维细胞生长因子和部分脱蛋白骨对兔股骨头缺损再血管化的影响

背景股骨头缺血坏死的治疗是骨科领域的一大难题.然而,近来的研究表明碱性成纤维细胞生长因子是一有效的血管生成因子,推测其在股骨头缺血坏死的治疗方面可能具有一定潜能.目的模拟临床上治疗股骨头坏死的开窗法的手术过程制备兔股骨头缺损,并植入碱性成纤维细胞生长因子和部分脱蛋白骨,以探讨碱性成纤维细胞生长因子对股骨头缺损的再血管化作用.设计随机对照实验.单位昆明医学院动物实验中心.材料实验于2002-07/2003-07在昆明医学院动物实验中心完成.成年雄性健康新西兰白兔27只,体质量2.2～2.8 kg.随机分为碱性成纤维细胞生长因子/部分脱蛋白骨复合骨组,单纯部分脱蛋白骨组,空白对照组,每组9只.方法①制备碱性成纤维细胞生长因子/部分脱蛋白骨复合骨含碱性成纤维细胞生长因子10 ng/mm3载体.②股骨头缺损模型的建立及修复取兔27只,在股骨头颈交界处开窗,建立骨缺损模型.复合骨组植入碱性成纤维细胞生长因子/部分脱蛋白骨;部分脱蛋白骨组植入单纯部分脱蛋白骨;空白对照组不植入任何植入物.实验动物分别于术后2,4,8周各时相点麻醉状态下进行墨汁灌注并处死,取材.主要观察指标①兔股骨头标本组织学检查及血管计数.②微血管面积图像分析.结果27只兔均进入结果分析.①兔股骨头标本组织学检查及血管计数术后8周,复合骨组移植材料被骨组织所取代,骨髓腔形成,其中有丰富的髓内血管存在;脱蛋白骨组移植材料亦被骨样组织包裹,且大部分部分吸收;空白对照组股骨头的缺损区被纤维组织填充,在缺损区周边的结缔组织中可见新生骨样组织和散在软骨细胞岛,血管数量较少.2周微血管计数复合骨组均显著高于部分脱蛋白骨组和空白对照组[(31.833±7.914)比(22.917±2.079)和(11.250±4.220)(血管数量/视野),P＜0.01,P＜0.05];4周和8周微血管计数复合骨组和脱蛋白骨组均显著高于空白对照组.②微血管面积图像分析20μm厚的透明标本光镜下观察见术后2,4,8周,复合骨组修复区血管丰富并吻合成网,部分脱蛋白骨组血管丰富吻合成网,空白组血管散在.结论碱性成纤维细胞生长因子具有促进股骨头缺损的再血管化作用,其在治疗股骨头坏死方面具备一定的潜能和优势.     

组织多普勒成像技术评价2型糖尿病患者股动脉远端血流阻断下股动脉壁弹性的价值

目的探讨组织多普勒成像技术评估2型糖尿病（T2DM）患者股动脉远端血流阻断下股动脉壁弹性的价值。 方法2012年12月至2014年2月绍兴第二医院收治T2DM患者64例,其中34例股动脉内膜中层厚度（IMT）<1.0 mm,30例股动脉IMT≥1.0 mm,同时选取同期30名健康体检者作为健康对照组。高频超声观察股动脉结构并测量股动脉IMT,应用脉冲多普勒（PW）技术测量股动脉血流频谱。使用血压机阻断股动脉远端血流后,应用PW及组织速度成像（TVI）技术测得股动脉后壁组织运动曲线,测量第一、二峰持续时间。采用t检验分别比较T2DM股动脉IMT<1.0 mm组患者、T2DM股动脉IMT≥1.0 mm组患者与健康对照组受试者IMT差异;采用秩和检验分别比较3组受试者股动脉远端血流阻断下股动脉后壁组织运动曲线第一、二峰持续时间差异。 结果健康对照组受试者、T2DM股动脉IMT<1.0 mm组患者、T2DM股动脉IMT≥1.0 mm组患者IMT分别为（0.80±0.09）、（0.82±0.08）、（1.26±0.19）mm。T2DM股动脉IMT<1.0 mm组患者与健康对照组受试者IMT差异无统计学意义（t=1.67,P=0.10）,T2DM股动脉IMT≥1.0 mm组患者IMT大于健康对照组受试者,且差异有统计学意义（t=16.88,P=0.00）。3组受试者股动脉远端血流阻断前股动脉血流频谱均为一高二低三相波型,股动脉远端血流阻断后股动脉血流频谱也均为二高二低四相波型。股动脉远端血流阻断下,健康对照组受试者、T2DM股动脉IMT<1.0 mm组患者、T2DM股动脉IMT≥1.0 mm组患者股动脉后壁组织运动曲线第一峰持续时间分别为（220.77±59.07）、（250.96±42.62）、（246.55±52.41）ms,3组间差异无统计学意义（u=3.258,P=0.196）;第二峰持续时间分别为（209.18±42.99）、（272.27±39.98）、（344.71±43.96）ms,3组间差异有统计学意义（u=74.150,P=0.000）。 结论股动脉后壁组织运动曲线第二峰持续时间可作为T2DM患者股动脉壁IMT改变前评价股动脉弹性的一个有效指标。