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1.
目的 探讨高机械指数超声辐照微泡对结肠癌细胞骨架的影响.方法 体外培养结肠癌细胞株(Lovo细胞),分为对照组、微泡+超声辐照组、单纯超声辐照组和单纯微泡组.使用超声造影剂SonoVue,探头频率1.5 MHz,机械指数1.7进行超声间歇辐照,激光共聚焦显微镜观察结肠癌Lovo细胞微丝、微管变化.结果 对照组微管染色表现为细胞致密的丝状网络结构,向细胞边缘呈放射性延伸;微泡+超声组微管表达较对照组减弱、稀疏,网络样结构主要沿细胞长轴排列.对照组细胞微丝染色表现为细胞致密网络状细丝样结构,向四周伸出许多细短的毛刺状突起,有明显的拉丝状感觉和方向性;微泡+超声组Lovo细胞胞质中部的网络状细丝明显减少,荧光暗淡,细短的毛刺状突起明显减少.单纯超声组与单纯超声微泡组微丝、微管均与对照组无明显差别.结论 高机械指数超声造影增强其空化效应能改变微丝、微管的组装和分布,对肿瘤细胞的侵袭、转移有一定的抑制作用.  相似文献   

2.
目的 探讨超声联合微泡对不同细胞周期血管平滑肌细胞(VSMCs)增殖和凋亡的影响.方法 组织贴块法培养大鼠胸主动脉VSMCs,采用血清饥饿法和胸苷双阻断法使细胞同步化, 并用流式细胞仪分析同步化结果.以频率1 MHz、声强0.3 W/cm2的连续波超声联合脂质微泡辐照不同细胞周期的VSMCs 120 s,分别用噻唑蓝(MTT)比色法、流式细胞仪AnnexinV/PI染色、免疫细胞化学技术检测VSMCs的增殖、凋亡和增殖细胞核抗原(PCNA)表达.结果 成功获得同步化的G0/G1期和S期VSMCs,其同步化率分别为89.53%和66.87%.超声联合微泡可显著抑制S期细胞的增殖并下调PCNA蛋白表达,对G0/G1期细胞的增殖和PCNA表达无明显影响;G0/G1期和S期细胞的凋亡率均较辐照前增高,且对于S期细胞超声联合微泡诱导凋亡的作用更为显著.结论 超声联合微泡对处于增殖状态的VSMCs具有显著的抑制增殖和诱导凋亡的作用.  相似文献   

3.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

4.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

5.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

6.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

7.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

8.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

9.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

10.
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.  相似文献   

11.
目的:通过观察粉防己碱对人结肠癌细胞株HT-29增殖与凋亡的影响,初步探讨粉防己碱对结肠癌的体外抗肿瘤效应。方法:采用MTT比色法观察粉防己碱对HT-29细胞的增殖抑制效应,利用细胞DNA琼脂糖凝胶电泳及细胞凋亡荧光染色法检测粉防己碱诱导肿瘤细胞凋亡的作用,以免疫细胞化学法检测粉防己碱对Bcl-2和BAX表达水平的影响。结果:MTT比色法显示粉防己碱对人结肠癌细胞株HT-29增殖有抑制作用,其抑制效应具有剂量依赖的特点,细胞凋亡荧光染色法、琼脂糖凝胶电泳表明粉防己碱可诱导HT-29细胞凋亡,免疫细胞化学法显示粉防己碱上调BAX基因表达,下调Bcl-2基因表达。结论:粉防己碱对人结肠癌细胞株HT-29增殖的抑制效应具有剂量依赖性,并可诱导细胞凋亡,其抗肿瘤效应可能与凋亡相关基因表达的调控有关。  相似文献   

12.
目的 评价超声辐照载血管内皮细胞生长因子抑制剂(vascular endothelial growth factor inhibitor,VEGFI)脂质微泡对结肠癌的治疗效果.方法 将稳定表达绿色荧光蛋白的结肠癌细胞系接种于Balb/c裸鼠皮下,建立结肠癌移植瘤模型.将48只皮下种植结肠癌移植瘤的Balb/c裸鼠随机分为4组:A组(12只),空白对照组,不接受超声造影及辐照;B组(12只),接受裸脂质微泡超声造影及假照;C组(12只),裸脂质微泡超声造影及超声辐照;D组(12只),载VFGFI脂质微泡及超声辐照.辐照前和3次辐照后第1d、第3d、第7d、第11d、第16d分别对各组裸鼠测量体质量和荧光摄片,测量肿瘤大小,计算肿瘤体积,绘制肿瘤体积生长曲线及裸鼠体质量生长曲线.16d后处死裸鼠,切除并分离肿瘤组织,测量各组肿瘤质量.结果 辐照前各组裸鼠体质量和肿瘤体积差异无统计学意义(P>0.05);随访期间,A组与B组肿瘤体积持续增大,两组间比较差异无统计学意义(P>0.05);辐照后第7d,C组及D组肿瘤体积均小于A组,差异有统计学意义(P<0.01),C组与D组之间比较差异无统计学意义(P>0.05);第7d后C组肿瘤体积变化不明显,到第16d肿瘤体积仍然小于A组,差异有统计学意义(P<0.01),而D组肿瘤体积进一步缩小,与C组比较差异有统计学意义(P<0.05).切除肿瘤组织,D组肿瘤质量低于C组(P<0.05),C组低于A组(P<0.05),而A、B组之间肿瘤质量差异无统计学意义(P>0.05).观察期间各组裸鼠体质量差异无统计学意义(P>0.05).结论 载VEGFI微泡联合超声辐照可以增强对结肠癌的治疗效果.  相似文献   

13.
目的 评价高机械指数实时灰阶谐波超声造影诊断肾动脉狭窄的临床价值.方法 怀疑肾动脉狭窄者21例,包括3例肾移植术后患者,行常规彩色多普勒超声和谐波造影检查.使用SonoVue造影剂,机械指数设置在1.0左右.超声检查结果与X线血管造影、CT血管成像、磁共振血管成像检查结果相对照.结果 常规超声诊断肾动脉狭窄的敏感性、特异性、阳性预测值、阴性预测值及准确性分别为85.7%,57.1%,80.0%,66.7%及76.2%;结合谐波造影可分别提高到100%,66.7%,88.2%,100%及90.5%.结论 高机械指数谐波超声造影能够明显提高肾动脉与周围组织的回声对比,直观显示肾动脉的流道变化,有助于对肾动脉狭窄的诊断.  相似文献   

14.
目的 应用超声造影评价超声介导的载血管内皮细胞生长因子抑制剂(vascular endothelial growth factor inhibitor,VEGFI)微泡对实验裸鼠结肠癌的治疗效果.方法 18只皮下种植结肠癌肿瘤的Balb/c裸鼠模型随机分为三组:A组(6只)为对照组,接受裸脂质微泡超声造影检查及假照;B组(6只),裸脂质微泡超声造影检查及超声辐照;C组(6只),载VEGFI脂质微泡及超声辐照.辐照前、辐照后1d、辐照后1周分别进行实时超声造影检查,存储图像进行声学定量脱机分析,获取峰值强度(PI)、局部血容量(RBV)、局部血流量(RBF)等造影参数并加以比较.结果 辐照前三组PI、RBV、RBF差异无统计学意义(P>0.05);A组假照后1 dPI、RBV、RBF高于假照前,但差异无统计学意义(P>0.05),1周后PI、RBV、RBF明显高于假照前,差异有统计学意义(P<0.05);B组辐照后1 dPI、RBF、RBV较辐照前减小,差异有统计学意义(P<0.05),而1周后PI、RBF、RBV又上升,与辐照前比较差异无统计学意义(P>0.05);C组辐照后1d、辐照后1周PI、RBV、RBF均较辐照前减小,差异有统计学意义(P<0.05).结论 载VEGFI微泡联合超声辐照可以增强对实验裸鼠结肠癌的治疗效果.  相似文献   

15.
目的采用正交实验设计法,优化低频(20kHz)低功率超声联合微泡诱导人雄激素非依赖型前列腺癌DUl45细胞早期凋亡的条件。方法以超声功率、微泡/细胞悬液体积比及超声辐照时间为正交优化的3个因素,每个因素设定3个水平,分别为超声功率60、80、100mW,微泡/细胞悬液体积比10%、20%、30%,超声辐照时间30、60、90S,根据三因素三水平设计正交实验表,得到9个实验组,按相应条件采用连续波辐照,辐照后细胞继续培养24h,采用流式细胞术检测细胞早期凋亡。应用正交优化得到的最优组合条件超声辐照实验组DUl45细胞,对照组细胞不予超声辐照,采用流式细胞术、透射电镜检测、观察实验组及对照组细胞早期凋亡。结果3个因素对细胞早期凋亡的影响程度为:超声功率〉微泡/细胞悬液体积比〉超声辐照时间。各个因素不同水平对细胞早期凋亡的影响程度分别为:超声功率:80mW〉60mW〉100mW,微泡/细胞悬液体积比:20%〉30%〉10%,超声辐照时间:60S〉90S〉30S。最优组合条件下,实验组细胞早期凋亡率为10.41%,对照组细胞早期凋亡率为O.94%。透射电镜下可见实验组细胞较对照组细胞的体积变小变圆,空泡增多,可见明显的凋亡小体形成。结论低频超声联合微泡诱导人雄激素非依赖型前列腺癌DUl45细胞早期凋亡的最优组合条件为:超声功率80mW,微泡/细胞悬液体积比20%,超声辐照时间60S。在此条件下实验组早期细胞凋亡率与对照组相比有明显差异。  相似文献   

16.
目的 总结不同临床分期宫颈癌的超声造影表现.方法 应用反向脉冲谐波超声造影技术,联合时间-强度曲线(TIC),分析48例经病理证实的不同临床分期宫颈癌患者的超声造影资料,总结宫颈癌超声造影增强方式.结果 宫颈癌超声造影表现为增强早期病变内造影剂灌注明显早于子宫肌层,呈均匀或不均匀增强;增强晚期造影剂消退早于子宫肌层,与正常宫体形成明显的界限,可清晰显示病变轮廓及对周围组织的浸润程度.结论 超声造影能较准确地判断宫颈癌分期及肿瘤浸润范围,在一定程度上弥补常规超声在宫颈癌诊断和分期中的不足.  相似文献   

17.
目的研究羟甲戊二酸单酰辅酶A(HMG-CoA)还原酶抑制剂——氟伐他汀在体外对人膀胱癌T24细胞凋亡、增殖和迁移及侵袭能力的影响,并探讨相关分子机制。方法 MTT法检测氟伐他汀对T24细胞增殖的影响;流式细胞术(PI和Annexin V双染法)检测细胞凋亡率的变化;划痕愈合和Transwell试验检测氟伐他汀对肿瘤细胞迁移和侵袭能力的影响;Western blot检测Bax、Cleaved-caspase-3等蛋白的表达情况。结果氟伐他汀在体外能显著抑制T24细胞的增殖,成明显浓度和时间依赖性;可以显著诱导肿瘤细胞凋亡;能显著抑制T24细胞的迁移和侵袭能力。结论氟伐他汀能显著诱导T24细胞发生凋亡,抑制肿瘤细胞的增殖、迁移和侵袭能力。  相似文献   

18.
目的探讨实时超声造影(CEUS)增强模式对胆囊癌的鉴别诊断价值。方法 20例胆囊癌和37例胆囊良性病变患者行常规超声及CEUS检查,重点观察病灶的造影增强模式。结果胆囊癌CEUS表现为病灶与胆囊壁同时显影,肝动脉期以增强明显为主,肝门静脉期以低增强为主。在CEUS增强早期呈高增强或等增强并在造影剂注射后35 s内变低增强者在胆囊癌中占95.0%(19/20);良性病变中占16.2%(6/37)(P=0.000)。病变表现为不均匀增强者在胆囊癌中占80.0%(16/20),良性病变中占23.3%(7/30)(P=0.000)。病变处胆囊壁完整性破坏者在胆囊癌中占85.0%(17/20),良性病变中占0.0%(0/37)(P=0.000)。超声检查诊断胆囊良恶性病灶的准确性92.98%、敏感性95.00%和特异性91.83%。结论 CEUS可提高超声诊断胆囊癌的准确率,能显著改善对胆囊疾病良恶性鉴别诊断能力,值得临床应用推广。  相似文献   

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