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Objective: To analyze the effect and mechanism of low intensity ultrasound on bone marrow suppression induced by docetaxel in rat models. Methods: Forty female rats were randomly divided into ultrasound group and control group (each n=20). Daily intraperitoneal injection of docetaxel (25 mg/kg) was given all rats for consecutive 4 days. Rats in ultrasound group were daily irradiated with low intensity ultrasound for consecutive 7 days after the first injection of docetaxel, while rats in control group received false irradiation at the same time. Routine blood examination and immunoglobulin tests were performed before docetaxel injection on the 4th, 7th and 14th day of irradiation. On the 4th day of irradiation, 4 rats in each group were randomly selected and killed for detection of stem cell factor (SCF), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Histological methods were used to detect the number of nucleated cells in the right femoral bone marrow tissue of rats before injection of docetaxel on the 4th, 7th and 14th day after the first time of irradiation. HE staining and pathological examinations of skin and muscle tissues of rats were performed on the 7th day after the first time of irradiation. Results: In ultrasound group, leukocyte, neutrophil and lymphocyte counts in peripheral blood on the 4th day after the first time of irradiatio, titer of immunoglobulin A (IgA) on the 7th and 14th day after the first time of irradiation, titers of immunoglobulin G (IgG) and immunoglobulin M (IgM) on the 4th and 14th day after the first time of irradiation increased obviously compared with those of control group (all P<0.05). The relative mRNA expression levels of SCF and ICAM-1 in bone marrow of rats in ultrasound group were significantly higher than those in control group, while the relative mRNA expression level of VCAM-1 in bone marrow of rats in ultrasound group was significantly lower than that in control group (all P<0.05). On the 4th and 7th day after the first time of irradiation, the numbers of bone marrow nucleated cells in ultrasound group were significantly higher than those in control group (both P<0.05). There was no obvious damage to the skin and muscle of the irradiated rats in both the two groups. Conclusion: Low intensity ultrasound is feasible in treatment of bone marrow suppression induced by docetaxel in rats, and its mechanism may be improvement of bone marrow hematopoietic microenvironment.
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AIM:To investigate the effect of low-intensity ultrasound combined with microbubble contrast agent on autophagic death of thyroid cancer cells,and to analyze the mechanism of autophagy activation and its effect on cell via-bility. METHODS:Human thyroid cancer cell line TPC1 was treated with low-intensity ultrasound at 20 kHz frequency and 80 mW intensity combined with microbubbles. The cell death and viability were analyzed by Live/Dead assay and CCK-8 assay 60,120 and 240 s after the treatment. The protein levels of microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ),autophagy-related protein 5 (ATG5) and SQSTM1/P62 were determined by Western blot. The number of in-tracellular autophagosomes was measured by the methods of monodansylcadaverine(MDC) staining,green fluorescent pro-tein (GFP)-LC3 transfection and transmission electron microscopy. The level of reactive oxygen species(ROS) was mea-sured and the effect of ROS on autophagy activation was evaluated by N-acetyl-L-cysteine (NAC) treatment. The effect of ATG5 siRNA transfection on autophagy was analyzed for determining the role of autophagic death. RESULTS:Low-intensi-ty ultrasound combined with microbubbles significantly promoted TPC1 cell death and inhibited TPC1 cell viability (P<0.05) in a time-dependent manner. Compared with low-intensity ultrasound group and microbubble group,ultrasound com-bined with microbubbles significantly increased the protein levels of LC3-Ⅱ and ATG5, but inhibited the protein level of P62 (P<0.05). The results of MDC staining,GFP-LC3 transfection and transmission electron microscopy showed that ul-trasound combined with microbubbles significantly increased the number of autophagosomes in the TPC1 cells. Compared with low-intensity ultrasound group and microbubble group, ultrasound combined with microbubbles increased the level of ROS,while NAC significantly reduced the protein level of LC3-Ⅱ (P<0.05). Thansfection with ATG5 siRNA inhibited the autophagy,significantly decreased the percentage of cell death and increased cell viability (P<0.05). CONCLU-SION:Low-intensity ultrasound combined with microbubbles promotes the autophagic cell death by increasing the level of ROS in thyroid cancer cells,leading to death of thyroid cancer cells.
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Sonodynamic therapy in many treatment methods of tumors, is a relatively new approach. Sonodynamic therapy as a treatment method is developed on the basis of photodynamic therapy. Compared to the lightwave, the ability to penetrate mammalian tissue of ultrasound is stronger. So sonodynamic therapy is a more promising therapeutic area. Many scholars have explored the clinical applications of sonodynamic therapy, and progress has been made in many areas. Because the female genitalia is connected the outside of body, it is more conducive that Sonodynamic therapy play a partial role. Sonodynamic therapy has some progress in the application of gynecology, and we hope it would have more progress in the treatment of endometrial disease.
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Objective To explore whether the low-intensity pulsed ultrasound (LIPU) could induce apoptosis on SMMC-7721 cells and to explore the underlying mechanism.Methods The SMMC-7721 cells were randomly divided into 4 groups:a blank control group,which was subject to sham exposure to ultrasound,and 3 ultrasound intervention groups exposed to ultrasound at intensities of 0.5,1.3 and 2.0 W/cm2,respectively.Then they were incubated for 6 h.The cell apoptosis,necrosis and changes of cell cycles were measured using the flow cytometry.The transmission electron microscope (TEM) was used to observe microstructural changes in the cells.The agarose gel electrophoresis (AGE) was used to examine the DNA fragmentation,and Western-blotting was employed to assess the protein expression of caspase-3.Results The average cell apoptosis rate of the 3 intervention groups were 4.66％,8.99％ and 32.41％,respectively.The percentage of cells in G2 phase increased significantly and those in G1 phase decreased significantly in the 3 intervention groups compared to the blank control group at the same time points.In the intervention groups,significant cell apoptosis was observed under TEM,and DNA ladders was seen in AGE,with DNA fragments appearing obviously when cells were incubated for 6 h and 9 h after ultrasound exposure.In intervention groups subject to 1.3 and 2.0 W/cm2 ultrasound exposure,the protein expression of caspase-3 was significantly higher than that of the control group.Conclusion LIPU can inhibit the proliferation and induce apoptosis of SMMC-7721 cells with a dose-dependent feature.The possible mechanism underlying the LIPU-induced cell apoptosis might be related to the activation of the mitochondria pathway,and especially the caspase-3 protein.
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Background and purpose:Low intensity ultrasound (LIUS) can kill cancer cells and promote their apoptosis. However, it is still unknown how it affects the migration and invasion of tumor cells. This study aimed to explore the effect of LIUS on human hepatocellular line MHCC97H in migration and metastasis and the possible mechanismin vitro.Methods:According to the intensity of ultrasonic irradiation, 4 experimental groups were established: control group (0 W/cm2), 0.5 W/cm2, 1.0 W/cm2 and 1.5 W/cm2group. The migration and invasion ability of hepatocellular cells was detected by scratch assay and Transwell migration and invasion assay after the irradiation of LIUS. The changes of cytoskeleton after irradiation were observed by microscope and F-actin green lfuorescence staining. The expressions of MMP-2 and MMP-9 were examined by real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot.Results:Low intensity ultrasound (≤1.5 W/cm2) promoted the migration and invasion of hepatocellular line MHCC97H. Scratch assay and Transwell assay showed much more cells under irradiation migrated through membrane than untreated. It was found that morphology of liver cancer cells changed after LIUS irradiation using optical microscope and lfuorescence microscope. The results of RTFQ-PCR and Western blot showed upregulation of MMP-2 expression by LIUS in MHCC97H and high expression of MMP-9 mRNA. Conclusion:Low intensity ultrasound may promote the migration and invasion of MHCC97H through changing cytoskeleton and upregulating protein expression of MMP-2.
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CONTEXTUALIZAÇÃO: A planimetria é um método utilizado para avaliar a evolução da cicatrização de feridas. A planimetria computacional é um método ainda em experimentação, mas cujas vantagens têm sido demonstradas em várias investigações. OBJETIVOS: Avaliar os efeitos do ultra-som pulsado de baixa intensidade sobre a cicatrização de lesão cutânea produzida na região dorsal de ratos, por meio da planimetria computacional. MATERIAIS E MÉTODOS: Utilizou-se 60 ratos machos Wistar (peso médio de 300g) divididos em dois grupos com 30 animais cada, de acordo com o tratamento: 1) irradiação simulada (controle); 2) irradiação efetiva (Freqüência fundamental de 1,5MHz, freqüência de repetição de pulsos de 1KHz, largura de pulso de 200µs, intensidade de 30mW/cm² SATA, dez minutos de aplicação em dias alternados). Cada grupo foi subdividido em três grupos, de acordo com o período de irradiação ultra-sônica, de três, sete e 14 dias, respectivamente, e a cicatrização foi avaliada por meio da planimetria, um decalque da lesão sendo obtido em papel especial, digitalizado e medido ao computador por meio de um programa gráfico. Análise estatística pelo método não-paramétrico de Mann-Whitney. RESULTADOS: Houve aumento significante (p<0,05) da área cicatrizada no grupo 2 (141,88±18,50mm²) em relação ao grupo 1 (117,38±15,14mm²), no 14º dia. Não houve diferenças significantes entre os grupos nos demais períodos. CONCLUSÕES: O ultra-som pulsado de baixa intensidade estimula a cicatrização cutânea por segunda intenção em condições experimentais. A planimetria computacional mostrou-se um recurso de baixo custo, fácil manuseio e de aplicabilidade clínica.
BACKGROUND: Planimetry is a method used to evaluate the progression of skin wound healing. Computerized planimetry is still an experimental method, but its advantages have been demonstrated in several investigations. OBJECTIVE: To evaluate the effects of low-intensity pulsed ultrasound on the healing of a skin lesion produced on the dorsal region of rats, by means of computerized planimetry. METHODS: Sixty male Wistar rats of mean weight 300g were used. They were divided into two groups according to the treatment applied: 1) simulated irradiation (control); 2) effective irradiation (fundamental frequency 1.5MHz, pulse repetition frequency 1KHz, pulse width 200µs, SATA intensity 30mW/cm² and application for ten minutes on alternate days). Each group was divided into three subgroups according to the length of time for which ultrasound irradiation was applied of three, seven and 14 days, respectively, and healing was evaluated by means of planimetry; a tracing of the wound was obtained on special paper and this was digitized and measured by means of a graphing software. Statistical analysis was performed using the Mann-Whitney non-parametric method. RESULTS: The healed area was significantly greater (p<0.05) in group 2 (141.88±18.50mm²) than in group 1 (117.38±15.14mm²) on the 14th day. There were no significant differences between the subgroups for the other experimental periods. CONCLUSIONS: Low-intensity pulsed ultrasound irradiation stimulated secondary skin healing under these experimental conditions. Computerized planimetry was shown to be a low cost method that was easy to use and present clinical applicability.
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@#Nonunion of longbones is a significant consequence in treating fractures, which is not easy to treat. Thes reviewed the basic knowledge and main progress regarding the cause, pathogenesis, pathophysiology of nonunion, and the methods that may be applied to the treatment of nonunion especially those non-operative one.
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In order to evaluate the efficacy of low intensity ultrasound and tissue engineering technique to repair segmental bone defects, the rabbit models of 1.5-cm long rabbit radial segmental osteoperiosteum defects were established and randomly divided into 2 groups. All defects were implanted with the composite of calcium phosphate cement and bone mesenchymal stem cells, and additionally those in experimental group were subjected to low intensity ultrasound exposure, while those in control group to sham exposure. The animals were killed on the postoperative week 4, 8 and 12 respectively, and specimens were harvested. By using radiography and the methods of biomechanics, histomorphology and bone density detection, new bone formation and material degradation were observed. The results showed that with the prolongation of time after operation, serum alkaline phosphatase (AKP) levels in both groups were gradually increased, especially in experimental group,reached the peak at 6th week (experimental group: 1.26 mmol/L; control group: 0.58 mmol/L), suggesting the new bone formation in both two group, but the amount of new bone formation was greater and bone repairing capacity stronger in experimental group than in control group. On the 4th week in experimental group, chondrocytes differentiated into woven bone, and on the 12th week, remodeling of new lamellar bone and absorption of the composite material were observed. The mechanical strength of composite material and new born density in experimental group were significantly higher than in control group, indicating that low intensity ultrasound could not only effectively increase the formation of new bone, but also accelerate the calcification of new bone. It was concluded that low intensity ultrasound could evidently accelerate the healing of bone defects repaired by bone tissue engineering.
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Objective:To study the influence of low intensity ultrasound on the contraction of uterine smooth muscle in rats in vivo. Methods:The uteri in rats in vivo were irradiated by low intensity ultrasound(0.8MHz,2W/cm2) for 10 min.The contraction frequency and amplitude were recorded with BL-410 biological experimental system. Results:It could be seen that the contraction frequency、amplitude、tension and contraction activity were significantly increased during ultrasonic irradiation(P
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PURPOSE: We investigated the effects of low intensity ultrasound on the co-culture of human osteoblastic cells with endothelial cells and analysed cell proliferation and growth factors that might be involved in bone formation. METHODS: Cell culture system was established with human osteoblastic cells (SaOS-2) and primary isolated human umbilical vein endothelial cells (HUVEC). Low intensity ultrasound (1 MHz) treatment was administered for 20 minutes per day to each well for 4 consecutive days and its effects were determined by analysing cell proliferation activity, alkaline phosphatse and amount of growth factors such as basic fibroblast growth factor, transforming growth factor beta and transforming growth factor beta in the conditioned medium. RESULTS: Low intensity ultrasound treatment increased cell proliferation activity, level of the basic fibroblast growth factor and transforming growth factor beta in the co-culture system. The levels of alkaline phosphatase and transforming growth factor beta in the conditioned medium were not changed. We could not observe the change of cell proliferation or amount of various growth factors in a culture system of SaOS-2 or HUVEC. CONCLUSION: The low intensity ultrasound treatment can increase osteogenesis by certain interaction between the two cells and increase in bFGF and TGF beta participate in the process.