Spatial cell fate manipulation of human pluripotent stem cells by controlling the microenvironment using photocurable hydrogel.

Human pluripotent stem cells (hPSCs) dynamically respond to their chemical and physical microenvironment, dictating their behavior. However, conventional in vitro studies predominantly employ plastic culture wares, which offer a simplified representation of the in vivo microenvironment. Emerging evidence underscores the pivotal role of mechanical and topological cues in hPSC differentiation and maintenance. In this study, we cultured hPSCs on hydrogel substrates with spatially controlled stiffness. The use of culture substrates that enable precise manipulation of spatial mechanical properties holds promise for better mimicking in vivo conditions and advancing tissue engineering techniques. We designed a photocurable polyethylene glycol-polyvinyl alcohol (PVA-PEG) hydrogel, allowing the spatial control of surface stiffness and geometry at a micrometer scale. This versatile hydrogel can be functionalized with various extracellular matrix proteins. Laminin 511-functionalized PVA-PEG gel effectively supports the growth and differentiation of hPSCs. Moreover, by spatially modulating the stiffness of the patterned gel, we achieved spatially selective cell differentiation, resulting in the generation of intricate patterned structures.

Grazing inhibition in Daphnia and Bosmina by colony formation of Desmodesmus subspicatus triggered by sodium octyl sulfate.

There are growing concerns that several aquatic contaminants can indirectly alter biological interactions by inhibiting the adaptive phenotypic plasticity of organisms, even at nonlethal concentrations. In Scenedesmaceae, a family of green algae, many chemicals interfere with defensive colony formation against grazers (i.e., through induced or limited coloniality). Although several studies have demonstrated that the effects of coloniality can limit the feeding capacity of Daphnia spp., grazing inhibition in other zooplankton species is not well understood. In this study, we examined the influence of sodium octyl sulfate (SOS) on the growth and morphology of Desmodesmus subspicatus and on the feeding rates of three cladoceran species (Daphnia galeata, Bosmina longirostris, and Bosmina fatalis) feeding on SOS-induced colonies under factorial conditions of different food levels and grazer ages. SOS remarkably induced colony formation with no observed effect on growth in D. subspicatus. D. galeata and B. fatalis showed a remarkable reduction in feeding rates when they fed on colonial D. subspicatus, whereas no significant effect of the prey morphotype was found on the feeding rates of B. longirostris. Microscopic observations of algal morphology after being grazed showed that each species can consume colonial prey depending on food level and age. Comparisons of the inhibition ratio of feeding among the three cladocerans revealed that Daphnia was more sensitive to prey coloniality compared with Bosmina. Our findings provide specific insights into the effects of chemically interfered colony formation on population dynamics and community structures.

Anaplastic Lymphoma Kinase Overexpression Is Associated with Aggressive Phenotypic Characteristics of Ovarian High-Grade Serous Carcinoma.

Deregulated full-length anaplastic lymphoma kinase (ALK) overexpression has been found in some primary solid tumors, but little is known about its role in ovarian high-grade serous carcinoma (HGSC). The current study focused on the functional roles of ALK in HGSC. Cytoplasmic ALK immunoreactivity without chromosomal rearrangement and gene mutations was significantly higher in HGSC compared with non-HGSC-type ovarian carcinomas, and was significantly associated with several unfavorable clinicopathologic factors and poor prognosis. HGSC cell lines stably overexpressing ALK exhibited increased cell proliferation, enhanced cancer stem cell features, and accelerated cell mobility, whereas these phenotypes were abrogated in ALK-knockdown cells. Expression of the nervous system-associated gene, ELAVL3, and the corresponding protein (commonly known as HuC) was significantly increased in cells overexpressing ALK. Expression of SRY-box transcription factor (Sox)2 and Sox3 (genes associated with the neural progenitor population) increased in ALK-overexpressing but not ALK-knockdown cells. Furthermore, overexpression of Sox2 or Sox3 enhanced both ALK and ELAVL3 promoter activities, suggesting the existence of ALK/Sox/HuC signaling loops. Finally, ALK overexpression was attributed to increased expression of neuroendocrine markers, including synaptophysin, CD56, and B-cell lymphoma 2, in HGSC tissues. These findings suggest that overexpression of full-length ALK may influence the biological behavior of HGSC through cooperation with ELAVL3 and Sox factors, leading to the establishment and maintenance of the aggressive phenotypic characteristics of HGSC.

Colony Formation in Three Species of the Family Scenedesmaceae (Desmodesmus subspicatus, Scenedesmus Acutus, Tetradesmus Dimorphus) Exposed to Sodium Dodecyl Sulfate and its Interference with Grazing of DAPHNIA Galeata.

By mimicking the info-chemicals emitted by grazers, the common anionic surfactant sodium dodecyl sulfate (SDS) can induce colony formation in the green algal genus Scenedesmus at environmentally relevant concentrations. The morphometric effects can hinder the feeding efficiency of grazers, reducing energy flow along the pelagic food chain from Scenedesmus to consumers. Despite this potential ecological risk, few studies exist on whether the SDS-triggered induction of colonies is common in other species of the family Scenedesmaceae. Here, we investigated the effects of SDS on the growth and morphology of three species of Scenedesmaceae (Desmodesmus subspicatus, Scenedesmus acutus, and Tetradesmus dimorphus) and on the clearance rates of Daphnia galeata grazing on the SDS-induced colonies. SDS triggered colony formation in all algal species at concentrations nonlethal to them (0.1-10 mg L-1) in 72 h; however, the induction levels of colony formation were generally lower than for those in the Daphnia culture medium. We also found that the SDS-induced colonial algae reduced D. galeata clearance rates. Our results highlight the potential effect of SDS on the Daphnia-Scenedesmaceae system by triggering the morphological response of Scenedesmaceae at concentrations below those that exert toxicity. Such disruptive effects of pollutants on predator-prey interactions should be considered within the framework of ecological risk assessments.

S100A4/Nonmuscle Myosin IIA/p53 Axis Contributes to Aggressive Features in Ovarian High-Grade Serous Carcinoma.

S100A4 is a small calcium-binding protein that exerts its biological functions by interacting with nonmuscle myosin IIA (NMIIA) and p53. Although S100A4 promotes metastasis in several tumors, little is known about its involvement in the progression of ovarian high-grade serous carcinomas (HGSCs). Herein, we focused on functional roles of the S100A4/NMIIA/p53 axis in these tumors. In HGSC cell lines harboring mutant p53, knockdown (KD) of S100A4 reduced the expression of several epithelial-mesenchymal transition/cancer stem cell markers and the ALDH1high population, consistent with an inhibition of stemness features. S100A4-KD also increased apoptosis, decreased cell proliferation, and accelerated cell mobility. This was accompanied by increased Snail expression, which, in turn, was likely due to loss of p53 function. In contrast, specific inhibition of NMIIA by blebbistatin induced phenotypes that-with the exception of cell proliferation and mobility-were opposite to those observed in S100A4-KD cells. In clinical samples, cytoplasmic and/or nuclear interactions between S100A4, NMIIA, and mutant p53 were observed. In addition, high expression of S100A4, but not NMIIA or p53, was a significant and independent unfavorable prognostic factor in HGSC patients. These findings suggest that, via its interaction with NMIIA and p53, overexpressed S100A4 may induce epithelial-mesenchymal transition/cancer stem cell properties in HGSC and elicit several other tumor-associated phenotypes.

Tumor growth suppression by the combination of nanobubbles and ultrasound.

We previously developed novel liposomal nanobubbles (Bubble liposomes [BL]) that oscillate and collapse in an ultrasound field, generating heat and shock waves. We aimed to investigate the feasibility of cancer therapy using the combination of BL and ultrasound. In addition, we investigated the anti-tumor mechanism of this cancer therapy. Colon-26 cells were inoculated into the flank of BALB/c mice to induce tumors. After 8 days, BL or saline was intratumorally injected, followed by transdermal ultrasound exposure of tumor tissue (1 MHz, 0-4 W/cm2 , 2 min). The anti-tumor effects were evaluated by histology (necrosis) and tumor growth. In vivo cell depletion assays were performed to identify the immune cells responsible for anti-tumor effects. Tumor temperatures were significantly higher when treated with BL + ultrasound than ultrasound alone. Intratumoral BL caused extensive tissue necrosis at 3-4 W/cm2 of ultrasound exposure. In addition, BL + ultrasound significantly suppressed tumor growth at 2-4 W/cm2 . In vivo depletion of CD8+ T cells (not NK or CD4+ T cells) completely blocked the effect of BL + ultrasound on tumor growth. These data suggest that CD8+ T cells play a critical role in tumor growth suppression. Finally, we concluded that BL + ultrasound, which can prime the anti-tumor cellular immune system, may be an effective hyperthermia strategy for cancer treatment.

Short- and long-term performance of risk calculation tools for mortality in patients with acute coronary syndrome.

Background: The mortality rate of acute coronary syndrome (ACS) remains high. Therefore, patients with ACS should undergo early risk stratification, for which various risk calculation tools are available. However, it remains uncertain whether the predictive performance varies over time between risk calculation tools for different target periods. This study aimed to compare the predictive performance of risk calculation tools in estimating short- and long-term mortality risks in patients with ACS, while considering different observation periods using time-dependent receiver operating characteristic (ROC) analysis. Methods: This study included 404 consecutive patients with ACS who underwent coronary angiography at our hospital from March 2017 to January 2021. The ACTION and GRACE scores for short-term risk stratification purposes and CRUSADE scores for long-term risk stratification purposes were calculated for all participants. The participants were followed up for 36 months to assess mortality. Using time-dependent ROC analysis, we evaluated the area under the curve (AUC) of the ACTION, CRUSADE, and GRACE scores at 1, 6, 12, 24, and 36 months. Results: Sixty-six patients died during the observation periods. The AUCs at 1, 6, 12, 24, and 36 months of the ACTION score were 0.942, 0.925, 0.889, 0.856, and 0.832; those of the CRUSADE score were 0.881, 0.883, 0.862, 0.876, and 0.862; and those of the GRACE score 0.949, 0.928, 0.888, 0.875, and 0.860, respectively. Conclusions: The ACTION and GRACE scores were excellent risk stratification tools for mortality in the short term. The prognostic performance of each risk score was almost similar in the long term, but the CRUSADE score might be a superior risk stratification tool in the longer term than 3 years.

Involvement of trigeminal axons in nose-to-brain delivery of glucagon-like peptide-2 derivative.

In our previous study, we created a glucagon-like peptide-2 (GLP-2) derivative with the functional sequence PAS-CPP to achieve efficient uptake by the respiratory epithelium and trigeminal nerve. By using octaarginine for cell penetrating peptides (CPP) and FFLIPKG, a reverse sequence of a part of the cathepsin D sequence for the penetration accelerating sequence (PAS), we found that the derivative was taken up by the cells through macropinocytosis and efficiently escaped from the endosomes and exited the cells. Moreover, it showed drug effects by intranasal (in.) administration at the same dose as intracerebroventricular (icv.) administration, which is direct drug administration into the brain. The purpose of this study was to elucidate the cause of the drug effect of in. administered PAS-CPP-GLP-2 at the same dose as that by icv. administration. The present results suggested that although icv. administered PAS-CPP-GLP-2 entered the cerebrospinal fluid, it barely penetrated the perivascular space of the brain, and therefore, only a small amount of the administered dose may have reached the site of action in the brain. In contrast, it was qualitatively suggested that in. administered PAS-CPP-GLP-2 migrates from the trigeminal nerve to the central nervous system via the principal sensory trigeminal nucleus and then through the trigeminal lemniscus. The present results show that nose-to-brain delivery by trigeminal axons, which is assumed to be a transcellular pathway, may be possible. As the drug can be delivered into the nerve, it is expected to be applied not only as a central delivery route but also for the treatment of neurological diseases.

The effect of positive autobiographical memory retrieval on decision-making under risk: A computational model-based analysis.

Psychiatric disorders such as depressive and anxiety disorders are associated with altered decision-making under risk. Recent advances in neuroeconomics and computational psychiatry have further discomposed risk-based decision-making into distinct cognitive computational constructs and showed that there may be disorder-specific alterations in these constructs. As a result, it has been suggested these cognitive computational constructs may serve as useful behavioral biomarkers for these disorders. However, to date, little is known about what psychological or behavioral interventions can help to reverse and manage the altered cognitive computational constructs underlying risk-based decision-making. In the present study, we set out to investigate whether recalling positive autobiographical memories may affect risk-based decision-making in healthy volunteers using a description-based task. Specifically, based on theories of behavioral economics, we dissected risk preference into two cognitive computational constructs, utility sensitivity and probability weighting. We found that compared to recalling neutral memories, retrieving positive autobiographical memories increased utility sensitivity (Cohen's d = 0.447), indicating reduced risk aversion. Meanwhile, we also tested the influence of memory retrieval on probability weighting, the effect, however, was unreliable and requires further in-depth investigation. Of clinical relevance, the change in risk aversion after recalling positive memories was in the opposite direction compared to those reported in psychiatric disorders. These results argue for the potential therapeutic effect of positive autobiographical memory retrieval for the amendment of altered risk-based decision-making in psychiatric disorders.

The Effect of Brief Stair-Climbing on Divergent and Convergent Thinking.

Recent studies show that even a brief bout of aerobic exercise may enhance creative thinking. However, few studies have investigated the effect of exercise conducted in natural settings. Here, in a crossover randomized controlled trial, we investigated the effect of a common daily activity, stair-climbing, on creative thinking. As experimental intervention, subjects were asked to walk downstairs from the fourth to the first floor and back at their usual pace. As control intervention, they walked the same path but using the elevator instead. Compared to using the elevator, stair-climbing enhanced subsequent divergent but not convergent thinking in that it increased originality on the Alternate Use Test (d = 0.486). Subjects on average generated 61% more original uses after stair-climbing. This is the first study to investigate the effect of stair-climbing on creative thinking. Our findings suggest that stair-climbing may be a useful strategy for enhancing divergent thinking in everyday life.

[Development of ultrasonic cancer therapy using ultrasound sensitive liposome].

Ultrasound (US) has been utilized as a useful tool for diagnosis and therapy. US mediated drug and gene delivery is paid to attention as a non-invasive system. The combination of US and microbubbles generated microjet stream by inducing disruption of bubbles and resulted in enhancing permeability of cell membrane. This phenomenon has been utilized as driving force for drug and gene delivery. Recently, we developed ultrasound sensitive liposome [Bubble liposome (BL)] containing perfluoropropane gas. US combined with BL could effectively transfer gene in vivo compared to conventional cationic liposomes. Using this method, we succeeded to obtain a therapeutic effect in cancer gene therapy with Interleukin-12 corded plasmid DNA. Therefore, it is expected that US combined with BL might be a useful non-viral vector system. From this result, the fusion of liposomal and ultrasound technologies would be important for establishment of advanced cancer therapy.

[Prediction of Wettability and Adhesion of Lotion to Skin Based on the OWRK Method].

In this study, we propose a new technique for evaluating wetting and adhesion of lotions to skin using surface tension measurements, contact angle measurements and calculations based on the Owens-Wendt-Rabel-Kaelble (OWRK) method. Three prescription lotions (Napageln® Lotion 3%, Sumilu® Lotion 3% and Felbinac Lotion 3% ï½¢Rakool｣） and two over-the-counter lotions (Feitas® Lotion and Salomethyl® FB Lotion α) were used. Based on the dispersive and polar components of the surface free energy of Yucatan micro pig (YMP) skin, isograms of contact angle (wetting envelope) and adhesion work of the YMP skin surface were constructed. Plotting the surface tension and its polar component of lotions on this isogram revealed that it is possible to predict the wettability and adhesion of lotions to YMP skin. Such diagrams can be easily constructed even using the surface free energy of other types of skin, such as that of humans and hairless mice. This evaluation method may be applicable to other external use medicines.

[Development of site specific gene delivery system with sonoporation].

In gene therapy, it is important to develop an effective and safe gene delivery system. Especially, from the viewpoint of reducing side effects, gene delivery into a specific site is essential. We previously, developed liposomal bubbles (Bubble liposomes) containing perfluoropropane. Bubble liposomes were useful as ultrasound enhanced gene delivery tools in vitro and in vivo. In this review, we introduced the characteristics of Bubble liposomes as ultrasound imaging agents and ultrasound enhanced gene delivery tools. Bubble liposomes worked as ultrasound imaging agents in cardiosonography. In addition, their combination with ultrasound exposure was able to deliver plasmid DNA in the femoral artery. The gene expression was only observed at the site of ultrasound exposure. Moreover, the gene delivery by Bubble liposomes and ultrasound exposure was more efficient than that by conventional lipofection method using Lipofectamine 2000. Therefore, it was suggested that Bubble liposomes might be a new class of tools for site specific gene delivery.

Spatiotemporal variability in a copepod community associated with fluctuations in salinity and trophic state in an artificial brackish reservoir at Saemangeum, South Korea.

Saemangeum Reservoir in South Korea is an estuarine system enclosed by a dyke construction, where seawater inflow and retained water outflow are managed by the opening/closing of sluice gates installed in the southern part of the dyke. An exchange of the reservoir water can cause spatiotemporal fluctuations in the salinity and trophic state, which are major drivers determining variation in the composition of biological communities in estuarine systems. Here, we investigated the seasonal and spatial variability in the copepod community and environmental conditions (water temperature, salinity, transparency, chlorophyll a concentration, total nitrogen, total phosphorus and Carlson's trophic state index) based on seasonally conducted field monitoring in the Saemangeum Reservoir from July 2013 to January 2018. In addition to the role of temperature, salinity and chlorophyll a concentration in structuring the copepod community and diversity, the biological indices of copepods with respect to salinity range and trophic state, were evaluated. The spatiotemporal variability in the salinity and trophic state variables showed contrasting patterns, and chlorophyll a concentration was negatively affected by salinity, indicating that the reservoir water was being highly exchanged with opening of the sluice gates. The mean trophic state index values, however, were constant in the eutrophic state (50-70). Dominant copepods were Acartia (A. hudsonica, A. sinjiensis, Acartia spp.) and Oithona (O. davisae and Oithona spp.), which are common species in eutrophic neritic water. Variation in the copepod community was mainly associated with the seasonal succession of the dominant species rather than a spatial gradient (from around the estuary to the sluice gates); however, site-specific differences in frequencies of several non-dominant species could be detected around the estuary (Sinocalanus tenellus) and the sluice gates (Centropages spp., Tigriopus spp. and Labidocera rotunda). The copepod diversity increased with species-richness from around the estuary to the sluice gates, which could result from variation in the site-specific location of non-dominant species. The frequency of particular species was also able to discriminate in terms of the salinity range (oligohaline: A. pacifica, S. tenellus and A. sinjiensis; mesohaline: Pseudodiaptomus inopinus; and polyhaline: C. abdominalis and Centropages spp.) and the trophic state (mesotrophic: C. abdominalis, Calanus sinicus and Centropages spp.; and hypereutrophic: S. tenellus, P. inopinus and Sinocalanus spp.). The findings from this study not only identify the factors determining spatiotemporal variation in the copepod community in the Saemangeum Reservoir, but also expand the applicability of copepods as biological indicators of conditions associated with salinity range and trophic state in other enclosed estuarine systems.

Efficient delivery of signal-responsive gene carriers for disease-specific gene expression via bubble liposomes and sonoporation.

Sonoporation is a promising method to intracellularly deliver synthetic gene carriers that have lower endocytotic uptake than viral carriers. Here, we applied sonoporation to deliver genes via polyethylene glycol (PEG)-grafted polymeric carriers that specifically respond to hyperactivated protein kinase A (PKA). PEG-grafted polymeric carrier/DNA polyplexes were not efficiently delivered into cells via the endocytotic pathway because of the hydrophilic PEG layer surrounding the polyplexes. However, the delivery of polyplexes into cells was significantly increased by sonoporation. The delivered polyplexes exhibited PKA-responsive transgene expression in PKA-overexpressing cells, but not in cells with low PKA activation. These results show that the sonoporation-mediated delivery of PEG-modified PKA-responsive polyplexes is a promising approach for safely applying gene therapy to abnormal cells with hyperactivated PKA.

Comparison of taxon-based and trophi-based response patterns of rotifer community to water quality: applicability of the rotifer functional group as an indicator of water quality.

Rotifer community is often used as a taxon-based bioindicator for water quality. However, studies of the planktonic community from the viewpoint of functional groups in freshwater ecosystems have been limited, particularly for rotifers. Because rotifers have various trophi types determining their feeding strategies, thereby representing an ecological niche, their functional feeding groups can act as biological and ecological indicators in lakes and reservoirs where planktonic communities are dominant. We analyzed the patterns of spatial distribution of the rotifer community in various reservoirs and then its relationship with water quality through redundancy and regression analyses. Compared with taxon-based composition, the response of trophi-based composition appears simplistic and showed clearer tendency in relation with water-quality variables. Each trophi responded differently by the degree of eutrophication indicating that each trophi group is possibly affected by environments such as the combinations of water-quality variables in different ways.

Nonviral gene delivery systems by the combination of bubble liposomes and ultrasound.

The combination of therapeutic ultrasound (US) and nano/microbubbles is an important system for establishing a novel and noninvasive gene delivery system. Genes are delivered more efficiently using this system compared with a conventional nonviral vector system such as the lipofection method, resulting in higher gene expression. This higher efficiency is due to the gene being delivered into the cytosol and bypassing the endocytosis pathway. Many in vivo studies have demonstrated US-mediated gene delivery with nano/microbubbles, and several gene therapy feasibility studies for various diseases have been reported. In addition, nano/microbubbles can deliver genes site specifically by the control of US exposure site. In the present review, we summarize the gene delivery systems by the combination of nano/microbubbles and US, describe their properties, and assess applications and challenges of US theranostics.

Development of fluorous lipid-based nanobubbles for efficiently containing perfluoropropane.

Nano-/microbubbles are expected not only to function as ultrasound contrast agents but also as ultrasound-triggered enhancers in gene and drug delivery. Notably, nanobubbles have the ability to pass through tumor vasculature and achieve passive tumor targeting. Thus, nanobubbles would be an attractive tool for use as ultrasound-mediated cancer theranostics. However, the amounts of gas carried by nanobubbles are generally lower than those carried by microbubbles because nanobubbles have inherently smaller volumes. In order to reduce the injection volume and to increase echogenicity, it is important to develop nanobubbles with higher gas content. In this study, we prepared 5 kinds of fluoro-lipids and used these reagents as surfactants to generate "Bubble liposomes", that is, liposomes that encapsulate nanobubbles such that the lipids serve as stabilizers between the fluorous gas and water phases. Bubble liposome containing 1-stearoyl-2-(18,18-difluoro)stearoyl-sn-glycero-3-phosphocholine carried 2-fold higher amounts of C3F8 compared to unmodified Bubble liposome. The modified Bubble liposome also exhibited increased echogenicity by ultrasonography. These results demonstrated that the inclusion of fluoro-lipid is a promising tool for generating nanobubbles with increased efficiency of fluorous gas carrier.

Systemic delivery of miR-126 by miRNA-loaded Bubble liposomes for the treatment of hindlimb ischemia.

Currently, micro RNA (miRNA) is considered an attractive target for therapeutic intervention. A significant obstacle to the miRNA-based treatments is the efficient delivery of miRNA to the target tissue. We have developed polyethylene glycol-modified liposomes (Bubble liposomes (BLs)) that entrap ultrasound (US) contrast gas and can serve as both plasmid DNA (pDNA) or small interfering RNA (siRNA) carriers and US contrast agents. In this study, we investigated the usability of miRNA-loaded BLs (mi-BLs) using a hindlimb ischemia model and miR-126. It has been reported that miR-126 promotes angiogenesis via the inhibition of negative regulators of VEGF signaling. We demonstrated that mi-BLs could be detected using diagnostic US and that mi-BLs with therapeutic US could deliver miR-126 to an ischemic hindlimb, leading to the induction of angiogenic factors and the improvement of blood flow. These results suggest that combining mi-BLs with US may be useful for US imaging and miRNA delivery.

[Novel dianostics and therapeutics with ultrasound technologies and nanotechnologies].

Ultrasound is a good tool for theranostics due to have multi-potency both of diagnostics with sonography and therapeutics with high intensity focused ultrasound (HIFU). In addition, microbubbles and nanobubbles are utilized as not only contrast imaging agent but also enhancer of drug and gene delivery by combination of ultrasound. Recently, we developed novel liposomal nanobubbles (Bubble liposomes) which were containing perfluoropropane. Bubble liposomes induced jet stream by low intensity ultrasound exposure and resulted in enhancing permeability of cell membrane. This phenomenon has been utilized as driving force for drug and gene delivery. On the other hand, the combination of Bubble liposomes and high intensity ultrasound induces strong jet stream and increase temperature. This condition can directly damage to tumor cells, we are applying this for cancer therapy. Therefore, their combination has potency for various cancer therapies such as gene therapy, immunotherapy and hyperthermia. In this review, we discuss about cancer therapy by the combination of Bubble liposomes and ultrasound.