H-22细胞声孔效应的应力阈值

理论和实验研究了超声空化场中的H-22型肝癌细胞产生可逆声孔效应的剪应力阈值.本文用1.37 MHz的聚焦声场,当超顺磁性纳米氧化铁在细胞悬液中的终浓度为410 μg/mL,换能器负载电功率为2 W,超声辐照60 s,细胞存活率90%以上时,有45.9±13.5%的细胞显示普鲁士蓝染阳性,暗示超声作用下,这些细胞表面曾出现可逆性微孔而使磁性微粒由此进入细胞内.利用无界自由空间微泡运动方程的球对称稳态解对实验条件下细胞膜表面的切变应力进行数值估算,结果表明,使H-22细胞产生可逆性声孔效应的微流剪应力阈值为697 Pa. 关键词： 声孔效应 磁性标记 微流 剪应力     

超声激活血卟啉对B16小鼠黑色素瘤细胞杀伤作用的研究初报

采用MTT法检测,经超声、超声 血卟啉分别作用后细胞线粒体内琥珀酸脱氢酶的活性,研究超声激活血卟啉对体外培养的B16小鼠黑色素瘤细胞的杀伤作用。结果表明:1.6MHz超声对B16小鼠黑色素瘤细胞辐照5min可表现出明显的杀伤效应,而超声协同被激活的血卟啉对B16细胞的杀伤作用比单纯超声的杀伤效应更加明显,且有显著性差异。     

超声作用可以提高公猪的生殖能力与质量

本实验研究了0.88和2.64 MHz高频超声对公猪精液质量的影响。采用的超声声强为0.05W/cm~2,辐照时间15、30s和1、2 min。超声波形分连续波和宽度为2、4、10ms的脉冲波。经超声处理过精子的活动性和绝对存活率分别增加到23％。最佳辐照时间30s而脉冲波宽度2ms。实验揭示了超声对猪的生理、生化特性和产仔的短期与长期影响。采用0.88MHz的超声辐照猪的生物活性点(BAP)和睾丸,精     

硼离子同位素位移的计算

应用Grasp92 程序包,计算了11,10B2+的22P3/2 →22S1/2 跃迁和22P1/2 →22S1/2 跃迁的同位素位移.22P3/2 →22S1/2 迁的同位素位移为29828 MHz,22P3/2 →22S1/2 跃迁的同位素位移为29840 MHz.这些理论结果与最新的实验结果是相符合的.     

空化水中HO2自由基的测定

采用频率为1．8MHz,声强为1－5W／cm2的超声波引发水中的空化效应,通过采用吡啶溶液作为HO2自由基捕获剂,测出了实验条件下空化水中HO2自由基的浓度水平为10－5M。     

双通道电光调Q射频激励波导CO2激光器研究

报道了双通道可调谐电光调Q射频激励波导CO2激光器,其中一通道是光栅选支电调Q的脉冲激光输出,调Q脉冲重复频率1Hz-10kHz可调,,脉冲峰值功率为150W,脉冲宽度为180ns,另一通道是光栅选支连续激光输出,可用压电陶瓷调节激光频率,同时分析了激光外差频率调谐范围,实验上获得最大脉冲激光外差频率调谐范围为150MHz。     

铝制焊接容器超声导波成像检测

针对中厚板铝制焊接容器的安全评价需要,论文研究了超声导波成像检测技术。通过对铝制焊接容器中不同模态超声导波的频散曲线及波结构分析,选择激励频率为2 MHz的S1模态作为中厚板铝制焊接容器健康监测的导波模态,并研制了适合铝制容器检测的2 MHz/S1模态压电晶片换能器,建立了超声导波成像系统。实验结果表明,该方法仅需经过有限次的扫查,就可以得到容器全壁厚范围内健康信息的超声导波图像,从而清晰地识别容器内的缺陷。该研究为中厚板铝制焊接容器的健康监测提供可行的技术方案。     

超声光栅实验中声致折射率变化的研究

为了研究声光效应中声致折射率变化情况,以超声光栅实验为基础,用CCD结合光强分析软件,分析了衍射光的强度分布;通过数值计算衍射光相对强度,比较得到了在超声场频率为10.27 MHz时,介质中的声光相位延迟Δ=1.2855 rad,声致折射率变化量Δn=3.014×10-6;进而讨论了电声换能器的机电耦合效率与超声频率的关系,给出当超声频率为10.33 MHz时,耦合效率最高,介质中折射率变化量最大;结果表明,在超声光栅实验中,引入这些测量内容,不仅能丰富实验内容,更能加深学生理解声光效应,扩展学生的知识面.     

光动力作用可致人肝癌细胞HepG2凋亡的实验研究

为探讨光动力作用（PDT）对体外培养的人肝癌细胞HepG2的杀伤效应，应用体外培养的人肝癌细胞HepG2，采用血卟啉衍生物（HPD）为光敏剂，用He-Ne激光器（波长632．8nm，能量密度120J／cm^2）进行激光照射，以系列浓度经不同剂量的光照后用MTT法测定PDT对肝癌细胞的相对抑制率，流式细胞仪检测其凋亡率．发现随光敏剂浓度的升高和照光剂量的增加，光动力作用对细胞的相对抑制率逐渐增大，在低光动力剂量下明显上升，随后上升渐趋缓慢达平台期．取在特定的光动力剂量下，肝癌细胞出现明显的凋亡现象，与对照组有明显的差异．实验结果表明，光动力作用下对体外培养的人肝癌细胞具有明确的杀伤效应，其机理可能与细胞凋亡有关．     

氯原子光诱导能级移动和增宽

应用半经典的微扰论方法计算出了氯原子光诱导能级移动和增宽，其大小与光强成正比。光频率为41018cm^-1时，氯原子初态3p^52P3/2^0和3p^52P1/2^0在非共振情况下，光移动分别为67．6和26．9MHz/Wμm^-2。当激光频率V接近3P^52P3／2^0→4s^2P3/2的跃迁频率时，氯原子基态3p^52P3/2^0的|MJ,MI>=|-1/2,3/2>→|1/2,3/2>超精细ESR谱线发生近共振频率光移动，大小量级为0．1MHz/Wμm^-2，能级增宽一般远小于光移动。     

The role of p53 in the response of tumor cells to sonodynamic therapy in vitro

p53 plays a pivotal role in apoptosis. In addition, p53 is currently extensively investigated as a promising strategy for highly specific anticancer therapy in chemotherapeutics and photodynamic therapy. However, the role of p53 in the response of tumor cells to sonodynamic therapy treatment is still unclear. In this study, we aim to investigate the activation of p53 in sonodynamic therapy. Three murine tumor models with distinct aggressiveness (S180, H-22 and EAC) were treated with 1.75 MHz continuous ultrasound at an acoustic intensity (I_SATA) of 1.4 W for 3 min in the presence of 20 μg/ml hematoporphyrin. The DNA fragment and nuclear damage were observed by TUNEL and single cell gel electrophoresis. Western blotting and RT-PCR were used to analyze the expression of p53, PUMA, Bax and Fas. Then we checked the translocation of p53 by confocal microscopy. DNA sequencing was used to determine the status of p53 gene in three tumor cell lines. Our results indicated that the level of p53 protein and mRNA increased significantly, and p53 activated the expression of its downstream pro-apoptosis gene PUMA, Bax and Fas in the S180 and H-22 cells. Meanwhile, p53 protein translocated onto mitochondria. In the EAC cells, expression and translocation of p53 was not found; the level of PUMA, Bax and Fas remained unaltered. The S180 cells showed most serious DNA fragment and nuclear damage with 77.43% TDNA; H-22 cells in the middle with 58.85% TDNA; whereas EAC cells appeared less nuclear material lost with just 15.82% TDNA. The results of DNA sequencing showed that the sequences of exons 5-8 of the p53 gene of S180, H-22 and EAC cells were the same with the sequences of wild-type p53 provided by NCBI. These results primarily demonstrated that: (1) p53 was activated to promote SDT-induced apoptosis through extrinsic and intrinsic signaling pathways in the S180 and H-22 cells; (2) cellular responses of different cells to SDT were distinct, the aggressive S180 cells were much more sensitive than H-22, whereas EAC cells were relatively less sensitive. The discrepancy among the cell lines may be due to different activation time of p53 protein.     

Comparisons among sensitivities of different tumor cells to focused ultrasound in vitro

This study is to test the sensitivities of different tumor cells to ultrasound irradiation at the frequency of 2.2 MHz for 60 s duration, and investigate the potential mechanism underlying different sensitivities. Three murine tumor models with distinct aggressiveness (S180, H-22 and EAC) were exposed to ultrasound to evaluate their sonodynamic efficiencies, and several biological parameters such as cell membrane permeability, lipid peroxidation (LPO), ultra-structure observation, intracellular reactive oxygen species (ROS) and mitochondria membrane potential (MMP) were analyzed after exposures. The results showed that cellular responses of different cells were distinct, of interest to note, the aggressive S180 cells were much more sensitive than others, whereas EAC cells were relatively more resistant to ultrasound irradiation. The direct comparisons among different types of cells indicate that the sono-sensitization seems to depend on the physiological and chemical properties of tumor cells. Perhaps sections of cell membrane became destabilized following the initial radical attack and LPO reaction, which caused S180 cells more susceptible to mechanical stresses during sonolysis. This study provides important implications for cancer therapy.     

Study of cell killing effect on S180 by ultrasound activating protoporphyrin IX

The present study was initiated to investigate the potential biological mechanism of cell killing effect on isolate sarcoma 180 (S180) cells induced by ultrasound activating protoporphyrin IX (PPIX). S180 cells were exposed to ultrasound for 30 s duration, at a frequency of 2.2 MHz and an acoustic power of 3 W/cm² in the presence of 120 μM PPIX. The viability of cells was evaluated using trypan blue staining. The generation of oxygen free radicals in cell suspensions was detected immediately after treatment using a reactive oxygen detection kit. A copper reagent colorimetry method was used to measure the level of FFAs released into cell suspensions by the process of cell damage induced by ultrasound and PPIX treatment. Oxidative stress was assessed by measuring the activities of key antioxidant enzymes (i.e., SOD, CAT, GSH-PX) in S180 tumor cells. Treatment with ultrasound and PPIX together increased the cell damage rate to 50.91%, while treatment with ultrasound alone gave a cell damage rate to 24.24%, and PPIX alone kept this rate unchanged. Colorimetry and enzymatic chemical methods showed that the level of FFAs in cell suspension increased significantly after the treatment, while the activity of all the above enzymes decreased in tumor cells at different levels, and were associated with the generation of oxygen free radicals in cell suspension after treatment. The results indicate that oxygen free radicals may play an important role in improving the membrane lipid peroxidation, degrading membrane phospholipids to release FFAs, and decreasing the activities of the key antioxidant enzymes in cells. This biological mechanism might be involved in mediating the effects on S180 cells and resulting in the cell damage seen with SDT.     

Experimental study on killing tumor cells by activation of hematoporphyrin derivatives by bi-frequency focal ultrasound in vitro

Tumor cells K562 were killed by the hematoporphyrin derivatives (HpD) activated by bi-frequency focal ultrasound in which a choice of irradiating parameter of ultrasound was tested by experiment. The effect of killing tumor cells was investigated by MTT method and compared with the contrast group. The results showed that the bi-frequency ultrasound exhibited an improved effect of killing tumor cells than single frequency ultrasound. It was 2-3 times higher than the single frequency ultrasound for the killing effect of tumor cells. After the irradiation of ultrasound, the cell-killing effect of hatching 16 h is better than that of hatching 4 h in the hatching tank. This might be due to delay of cell apoptosis.     

In vitro activation of mitochondria-caspase signaling pathway in sonodynamic therapy-induced apoptosis in sarcoma 180 cells

Sonodynamic therapy (SDT) has been shown to mediate apoptosis in many experimental systems, but the detailed mechanism of this process is unclear. In this study, we aim to investigate the potential participation of the mitochondria-caspase signaling pathway in the SDT-induced apoptosis in isolated sarcoma 180 (S180) cells. The cell suspension was treated with 1.75 MHz continuous ultrasound (US) at an acoustic intensity (I_SATA) of 1.4 W for 3 min in the absence or presence of 20 μg/ml hematoporphyrin (Hp). At different times after the SDT-treatment, the apoptotic cells were identified under a scanning electron microscope, and the apoptosis index (AI) was determined by flow cytometry. In addition, the mitochondrial membrane potential, permeabilization of the inner mitochondrial membrane, and translocation of apoptosis-related proteins were assessed by confocal microscopy. Simultaneously, the activation of some special apoptosis-associated proteins [caspase-9, caspase-3, polypeptide poly (ADP-ribose) polymerase (PARP), and Bax] was evaluated by western blotting. Our results indicate that the ultrasonically activated Hp can cause obvious cell apoptosis (AI, 57.66%) at 3 h after treatment, and this effect can be significantly reduced by caspase-9 inhibitor (AI, 20.76%) and the oxygen scavenger NaN₃ (20.11%). However, the apoptosis induced by ultrasound alone was relatively lower (28.33%) and was not reduced by NaN₃. Further, SDT caused an 82.1% reduction in the mitochondrial membrane potential and a 70.7% reduction in the permeabilization of the inner mitochondrial membrane immediately after treatment, and these two effects were obviously prevented by NaN₃. In comparison with the control cells, the SDT-treated cells showed obvious cytochrome-c and Bax translocations, caspase activation, Bax expression, and PARP cleavage at 1 h after SDT-treatment. However, in the cells treated with ultrasound alone, these phenomena partially and weakly occurred 3 h after exposure. These results primarily showed that the mitochondria-caspase signaling pathway in S180 cells was activated in the US- and SDT-induced apoptosis. Moreover, Hp significantly accelerates the process of apoptosis and enhances the cytotoxic effect of ultrasonic treatment. Singlet oxygen may be responsible for the mitochondrial damage and the activation of the apoptotic signaling pathway.     

Ultrasound-assisted photochemical oxidation of unsymmetrically substituted 1,4-dihydropyridines

The combination of ultrasound and photochemistry has been used for the oxidation of unsymmetrical 1,4-dihydropyridines to the pyridine derivatives. An ultrasonic probe of 24 kHz frequency and a Hg-lamp of 100 W have been used for this study. The effect of parameters such as ultrasonic intensity, the presence of oxygen and argon atmospheres and also the separate usage of one of these irradiation sources have been studied. Whereas sonication of these compounds alone did not result in the oxidation of them, the use of ultrasound increases the rate of photo-oxidation. The presence of oxygen decreases or increases the rate of reaction, depending on the type of excited state of 1,4-dihydropyridines involved in the reaction.     

Sonodynamically induced antitumor effect of hematoporphyrin on Hepatoma 22

The ultrasonically induced cytotoxic effects of hematoporphyrin (Hp) on Hepatoma 22 (H22) cells in vitro and vivo were investigated. Tumor cells were suspended in saline and exposed to ultrasound at 1.43 MHz for up to 60s in the presence and absence of Hp. The viability of cells was evaluated by trypan blue exclusion test. The ultra-structure changes of H22 cells induced by ultrasonic irradiation were evaluated by scanning electron microscope (SEM) and transmission electron microscope (TEM). Lipid peroxidation in cell was estimated by the thiobarbicturic acid (TBA) method. Our experiments indicated that the ultrasonic intensity of 2 W/cm(2), the Hp concentration of 100 microg/ml and the ultrasound exposure time of 60s were the best conditions for sonodynamic treatment in vitro. The tumor volume and weight after the combination of Hp with ultrasound were remarkably inhibited. SEM and TEM observation found the cell ultra-structure was significantly damaged, and lipid peroxidation level remarkably increased after sonodynamic treatment. This study suggested the ultra-structural changes may play a key role in cell destruction induced by sonodynamic treatment and the biological mechanism might be involved in mediating the killing effect on H22 cells in our experiment mode.     

Synergistic bactericidal effects and mechanisms of low intensity ultrasound and antibiotics against bacteria: a review

Low intensity ultrasonic therapy is always an important research area of ultrasonic medicine. This review concentrates on low intensity ultrasound enhancing bactericidal action of antibiotics against bacteria in vitro and in vivo, including planktonic bacteria, bacterial biofilms, Chlamydia, and bacteria in implants. These literatures show that low intensity ultrasound alone is not effective in killing bacteria, while the combination of low intensity ultrasound and antibiotics is promising. Low intensity ultrasound facilitating antibiotic treatment is still in its infancy, and still requires a great deal of research in order to develop the technology on medical treatment scale.     

Instant magnetic labeling of tumor cells by ultrasound in vitro

Magnetic labeling of living cells creates opportunities for numerous biomedical applications. Here we describe an instantly cell magnetic labeling method based on ultrasound. We present a detailed study on the ultrasound performance of a simple and efficient labeling protocol for H-22 cells in vitro. High frequency focus ultrasound was investigated as an alternative method to achieve instant cell labeling with the magnetic particles without the need for adjunct agents or initiating cell cultures. Mean diameter of 168 nm dextran-T40 coated superparamagnetic iron oxide (SPIO) nanoparticles were prepared by means of classical coprecipitation in solution in our laboratory. H-22 tumor cells suspended in phosphate-buffered saline (PBS, pH=7.2) were exposed to ultrasound at 1.37 MHz for up to 120 s in the presence of SPIOs. The cellular uptake of iron oxide nanoparticles was detected by prussion blue staining. The viability of cells was determined by a trypan blue exclusion test. At 2 W power and 60 s ultrasound exposure in presence of 410 μg/ml SPIOs, H-22 cell labeling efficiency reached 69.4±6.3% and the labeled cells exhibited an iron content of 10.38±2.43 pg per cell. Furthermore, 95.2±3.2% cells remained viable. The results indicated that the ultrasound protocol could be potentially applied to label cells with large-sized magnetic particles. We also calculated the shear stress at the 2 W power and 1.37 MHz used in experiments. The results showed that the shear stress threshold for ultrasonically induced H-22 cell reparable sonoporation was 697 Pa. These findings provide a quantitative guidance in designing ultrasound protocols for cell labeling.