Stereoselective Synthesis and Characterization of Indenone Azine-Based Electron-Accepting π-Conjugated Systems.

Indenone azines, in which the exocyclic C=C bond in dibenzopentafulvalene is replaced by an azine moiety (C=N-N=C), have been synthesized as novel electron-accepting π-conjugated scaffolds. Structural modulation at the 7,7'-positions of indenone azines enabled stereoselective syntheses of diastereomers in which the configurations of the two C=N bonds are E,E or Z,Z. X-ray crystallographic analyses revealed that all the indenone azines exhibit high coplanarity in contrast to the twisted frameworks of dibenzopentafulvalene derivatives, resulting in the formation of densely π-stacked structures. Electrochemical measurements and quantum chemical calculations revealed the electron-accepting character of indenone azines comparable to isoindigo dyes. In particular, the intramolecular hydrogen bonds of 7,7'-dihydroxy-substituted derivatives impart enhanced electron-accepting character and significantly red-shifted photoabsorption. This study demonstrates that indenone azines represent a promising candidate as electron-accepting building blocks for optoelectronic materials.

Stereoselective Synthesis and Characterization of Indenone Azine-Based Electron-Accepting π-Conjugated Systems.

Invited for the cover of this issue is the group of Aiko Fukazawa at Kyoto University. The image depicts a N-N component replacing the one originally located between the exocyclic C=C bond of a cross-conjugated dibenzofulvalene. Read the full text of the article at 10.1002/chem.202300181.

Gas Adsorption and Diffusion Behaviors in Interfacial Systems Composed of a Polymer of Intrinsic Microporosity and Amorphous Silica: A Molecular Simulation Study.

We investigate the adsorption and diffusion behaviors of CO2, CH4, and N2 in interfacial systems composed of a polymer of intrinsic microporosity (PIM-1) and amorphous silica using grand canonical Monte Carlo (GCMC) and molecular dynamics (MD) simulations. We build model systems of mixed matrix membranes (MMMs) with PIM-1 chains sandwiched between silica surfaces. Gas adsorption analysis using GCMC simulations shows that gas molecules are preferentially adsorbed in microcavities distributed near silica surfaces, resulting in an increase in the solubility coefficients of CO2, CH4, and N2 compared to bulk PIM-1. In contrast, diffusion coefficients obtained from MD simulations and then calibrated using the dual-mode sorption model show different tendencies depending on gas species: CO2 diffusivity decreases in MMMs compared to PIM-1, whereas CH4 and N2 diffusivities increase. These differences are attributed to competing effects of silica surfaces: the emergence of larger pores as a result of chain packing disruption, which enhances gas diffusion, and a quadrupole-dipole interaction between gas molecules and silica surface hydroxyl groups, which retards gas diffusion. The former has a greater impact on CH4 and N2 diffusivities, whereas the latter has a greater impact on CO2 diffusivity due to the strong quadrupole-dipole interaction between CO2 and surface hydroxyls. These findings add to our understanding of gas adsorption and diffusion behaviors in the vicinity of PIM-1/silica interfaces, which are unobtainable in experimental studies.

Photoinduced binding of malachite green copolymer to parallel G-quadruplex DNA.

Designing ligands that selectively target G-quadruplex DNAs has gained attention due to their possible roles in regulation of gene expression and as anti-cancer agents. In this article, we report irradiation-induced ligand binding to G-quadruplex DNAs which offers a novel approach to targeting specific G-quadruplexes. Photoinduced binding to G-quadruplex DNAs was observed for copolymers of poly(vinyl alcohol) carrying a malachite green moiety (PVAMG). This molecule has an aromatic ring with cationic charge, which after irradiation becomes a binding site for G-quadruplex DNA. PVAMGs acted as neutral polymers with no binding affinity under dark conditions. The photoinduced binding was revealed by fluorescence spectroscopy, NMR spectroscopy, UV melting curve, and DNA polymerase stop assay. PVAMGs showed preference to parallel G-quadruplex structures over mixed parallel/antiparallel structures. PVAMGs were found to be noncytotoxic under both dark and irradiated conditions up to a concentration of 20 µM.

Platelets: Small in Size But Essential in the Regulation of Vascular Homeostasis - Translation From Basic Science to Clinical Medicine.

Platelets are small blood cells that adhere to the site of vessel injury where von Willebrand factor (VWF) is expressed. Platelets bind to VWF through interaction with a membrane protein, glycoprotein (GP) Ibα. Next, the accumulated platelets are activated to change their morphological and biochemical characteristics. Various vasoactive substances, such as immune-regulatory CD40 ligand, are released locally from activated platelet cells to maintain homeostasis of the vascular system. Major roles played by platelets in the regulation of hemostasis and thrombus formation include local activation of the coagulation cascade. Translocation of negatively charged phospholipids to the surface of activated platelets helps in the formation of prothrombinase complex, which efficiently produces thrombin. Thrombin produces fibrin around the activated platelets and further activates the platelets through thrombin receptor stimulation. Of the various platelet-stimulating receptors and activation signals, cyclo-oxygenase-1, P2Y12 adenosine 5'-diphosphate receptor, and thrombin receptor (protease activated receptor)-1 blockers are used clinically as antiplatelet agents. In the future, precise understanding of the quantitative contribution of platelet function in hemostasis and pathological thrombus formation should lead to the development of effective antithrombotic agents without increasing the risk of serious bleeding complications.

Numerical Study of Cerebroarterial Hemodynamic Changes Following Carotid Artery Operation: A Comparison Between Multiscale Modeling and Stand-Alone Three-Dimensional Modeling.

Free outflow boundary conditions have been widely adopted in hemodynamic model studies, they, however, intrinsically lack the ability to account for the regulatory mechanisms of systemic hemodynamics and hence carry a risk of producing incorrect results when applied to vascular segments with multiple outlets. In the present study, we developed a multiscale model capable of incorporating global cardiovascular properties into the simulation of blood flows in local vascular segments. The multiscale model was constructed by coupling a three-dimensional (3D) model of local arterial segments with a zero-one-dimensional (0-1-D) model of the cardiovascular system. Numerical validation based on an idealized model demonstrated the ability of the multiscale model to preserve reasonable pressure/flow wave transmission among different models. The multiscale model was further calibrated with clinical data to simulate cerebroarterial hemodynamics in a patient undergoing carotid artery operation. The results showed pronounced hemodynamic changes in the cerebral circulation following the operation. Additional numerical experiments revealed that a stand-alone 3D model with free outflow conditions failed to reproduce the results obtained by the multiscale model. These results demonstrated the potential advantage of multiscale modeling over single-scale modeling in patient-specific hemodynamic studies. Due to the fact that the present study was limited to a single patient, studies on more patients would be required to further confirm the findings.

Molecular beam study of the scattering behavior of water molecules from a graphite surface.

Gas flow in nanospaces is greatly affected by the scattering behavior of gas molecules on solid surfaces, resulting in unique mass transport properties. In this paper, the molecular beam scattering experiment of water molecules on a graphite surface was conducted to understand their scattering dynamics in an incident energy range that corresponds to their thermal velocity distribution at room temperature (35-130 meV). Because of the large adsorption energy (∼100 meV), the scattering behavior is quite sensitive to the incident energy even within this narrow energy range. For relatively large incident energies, the direct-inelastic and trapping-desorption channels have comparable contributions to the scattering process on the surface at 300 K. In contrast, when the incident energy decreases well below the adsorption energy on the surface, the trapping-desorption channel becomes dominant, changing the scattering pattern from directional to diffusive scattering. As a result, the tangential momentum accommodation coefficient (TMAC), which significantly impacts the mass transport in nanospaces, largely depends on the incident energy. A decrease in the incident energy from 130 to 35 meV doubles the TMAC (0.42 to 0.86). In addition to the incident energy, the TMAC shows a strong dependence on the surface temperature. With increasing the surface temperature from 300 to 500 K, the scattering becomes more directional because of the increasing contribution of the direct-inelastic channel, which reduces the TMAC for the incident beam energy of 35 meV to 0.48.

Microbubble behavior in an ultrasound field for high intensity focused ultrasound therapy enhancement.

The enhancement of heating due to inertial cavitation has been focused to reduce the long treatment time of conventional high-intensity focused ultrasound (HIFU) therapy. The influences of the physical properties of surrounding tissues, initial void fraction, and spatial distribution of bubbles on microbubble-enhanced HIFU are examined. A bubble dynamics equation based on the Keller-Miksis equation is employed in consideration of the elasticity of surrounding tissue. The mixture phase and bubbles are coupled by the Euler-Lagrange method to take into account the interaction between ultrasound and bubbles. As a result, the temperature around the target increases with the initial void fraction. But at the high void fraction of 10(-5), ultrasound is too attenuated to heat the target, and the heating region moves to the transducer side. On the other hand, both the viscosity and shear elasticity of the surrounding media reduce the attenuation of ultrasound propagation through the bubbly mixture. Numerical results show that localized heating is induced with increasing viscosity or shear elasticity, though it depends on the pressure amplitudes. In addition, it was numerically confirmed that the localization of the microbubble distribution is important to obtain efficient localized heating.

Deep learning for hetero-homo conversion in channel-domain for phase aberration correction in ultrasound imaging.

Echo imaging in ultrasound computed tomography (USCT) using the synthetic aperture technique is performed with the assumption that the speed of sound is constant in the system. However, tissue heterogeneity causes a mismatch between the predicted arrival time and the actual arrival time of the echo signal, which will result in phase aberration, leading to the quality degradation of the reconstructed B-mode image. The conventional correction methods that use the correlation of each different channel require the presence of strong point scatterers and involve the problem of local solutions due to excessive correction. In this study, we propose a novel approach to correcting the signal distortion due to sound speed heterogeneity using a deep neural network (DNN). The DNN was trained to convert the distorted radio frequency (RF) inputs for the heterogeneous medium to the distortion-free RF outputs for the homogeneous medium. The network with U-net architecture using ResNet-34 as a backbone was trained using the hetero-homo corresponding channel-domain RF data generated via numerical simulations. The trained network performed phase aberration correction in the channel-domain RF, with the B-mode images reconstructed with the corrected RF demonstrating a higher contrast and an improved resolution compared with uncorrected cases. It was also demonstrated that the DNN model is robust to both varied reflection intensities and varied sound speed heterogeneities. The successful results demonstrated that the proposed DNN-based method is effective for phase aberration correction in US imaging.

Flattened 1D fragments of fullerene C60 that exhibit robustness toward multi-electron reduction.

Fullerenes are compelling molecular materials owing to their exceptional robustness toward multi-electron reduction. Although scientists have attempted to address this feature by synthesizing various fragment molecules, the origin of this electron affinity remains unclear. Several structural factors have been suggested, including high symmetry, pyramidalized carbon atoms, and five-membered ring substructures. To elucidate the role of the five-membered ring substructures without the influence of high symmetry and pyramidalized carbon atoms, we herein report the synthesis and electron-accepting properties of oligo(biindenylidene)s, a flattened one-dimensional fragment of fullerene C60. Electrochemical studies corroborated that oligo(biindenylidene)s can accept electrons up to equal to the number of five-membered rings in their main chains. Moreover, ultraviolet/visible/near-infrared absorption spectroscopy revealed that oligo(biindenylidene)s exhibit enhanced absorption covering the entire visible region relative to C60. These results highlight the significance of the pentagonal substructure for attaining stability toward multi-electron reduction and provide a strategy for the molecular design of electron-accepting π-conjugated hydrocarbons even without electron-withdrawing groups.

Ultrasound-mediated gene transfection in vitro: effect of ultrasonic parameters on efficiency and cell viability.

Ultrasound (US)-mediated gene transfection in the presence of microbubbles is a recently developed and promising non-viral gene delivery method. Optimising the parameters used in ultrasonic transfection is urgently required in order to realise higher transfection efficiencies in clinical settings. This study examined the effect of ultrasound exposure parameters on plasmid DNA transfection in mouse embryonic fibroblast cell lines using perfluorobutane bubbles. Variations in US intensity (0-11 W/cm2), pulse repetition frequency (PRF, 50-50,000 Hz), duty ratio (10 to 50%), exposure time (0-120 s) and microbubble volume concentration (0 to 10%) were tested, and the microbubble volume concentration was also monitored during exposure. Through the experiments, the mechanism of how variations in parameters influence US-mediated gene transfection was discussed, which can provide a basis for future applications of ultrasound mediated transfection.

Localized motion imaging for monitoring HIFU therapy: Comparison of modulating frequencies and utilization of square modulating wave.

High-intensity focused ultrasound (HIFU) has been successfully used as a minimally invasive cancer therapy method. For monitoring the therapy, the amplitude-modulated (AM) localized motion imaging (LMI) method had been proposed. This paper compares the performance of AM-LMI while using different sine modulating wave frequencies and proposes the utilization of square modulating waves to gain the advantages of both high and low modulating frequencies. A single element therapy transducer with a 2 MHz central frequency was driven by sine modulating waves with different frequencies (approximate 34, 67, 102, 168, and 201 Hz) and by square modulating waves with two frequencies (34 and 67 Hz). An imaging probe with a 5 MHz central frequency and a 20 MHz sampling frequency was mounted in the center hole of the therapy transducer to acquire pulse-echo data, which were used to estimate the tissue oscillation amplitude induced by the acoustic radiation force of the HIFU beam. The decrease ratio of the oscillation amount was then utilized to estimate the coagulated lesion length during the therapy. The comparison of modulating frequencies demonstrated that a higher frequency could bring higher sensitivity to small lesions, while a lower frequency not only gives greater noise robustness but also promotes the ability to estimate lengths of larger lesions. The utilization of a square modulating wave demonstrated its utility to produce tissue oscillation with multiple frequencies and gain the advantages of both high and low modulating frequencies.

Numerical Study on a Focus-Control Method Using Breast Model With Intentionally Assigned High-Absorbing Layer Near Skin for High-Intensity Focused Ultrasound Treatment.

To prevent undesirable skin burns that occur in high-intensity focused ultrasound (HIFU) treatment, we numerically study focus-control methods, such as phase compensation (PC) and amplitude adaptation (AA). We intentionally assign a high-absorbing layer (HAL) near the part of the skin, where heat generation and tissue ablation are observed, because of high energy loss in the interface between water and breast skin. Results show that PC improves the effectiveness of focusing by enhancing the focal peak and reducing the focal deviation; however, PC does not suppress skin burn. AA and PC eliminate skin burns only if appropriate amplitude weights are applied. A preliminary discussion on three algorithms for obtaining amplitude weights is conducted as follows; First, we switched off transducer channels using distance-to-HAL. This algorithm eliminates skin burns while causing other undesirable burns by preserving 100% input energy. Second, we use cross-correlated amplitude weights. It eliminates skin burn after properly limiting large-amplitude weights while producing focal necrosis in a smaller and slower manner. Third, we introduced root-mean-square (rms) level of back-propagated wave (BPW) into cross-correlated amplitude weights. This new algorithm produces focal ablation in 20 s without causing any skin burn. Although longer irradiation time brings back skin burn, the result is satisfying since short irradiation time is needed in HIFU treatment to avoid exceeding the physical endurance of human patients. Moreover, this work indicates that focus-control associated with an acoustic peak is insufficient. The effects of the high attenuation area are significant and should be captured.

Important Regulatory Roles of Erythrocytes in Platelet Adhesion to the von Willebrand Factor on the Wall under Blood Flow Conditions.

The role of erythrocytes in platelet adhesion to von Willebrand factor (VWF) on the vessel wall through their membrane glycoprotein (GP)Ibα under blood flow conditions has not yet been elucidated. Blood specimens containing fluorescent-labeled platelets and native, biochemically fixed, or artificial erythrocytes at various hematocrits were perfused on the surface of VWF immobilized on the wall at a shear rate of 1,500 s-1. The rates of platelet adhesion were measured under each condition. The computer simulation of platelet adhesion to the VWF on the wall at the same shear rate was conducted by solving the governing equations with a finite-difference method on a K computer. The rates of platelet adhesion were calculated at various hematocrit conditions in the computational domain of 100 µm (x-axis) × 400 µm (y-axis) × 100 µm (z-axis). Biological experiments demonstrated a positive correlation between the rates of platelet adhesion and hematocrit values in native, fixed, and artificial erythrocytes. (r = 0.992, 0.934, and 0.825 respectively, p < 0.05 for all). The computer simulation results supported the hematocrit-dependent increase in platelet adhesion rates on VWF (94.3/second at 10%, 185.2/second at 20%, and 327.9/second at 30%). These results suggest that erythrocytes play an important role in platelet adhesion to VWF. The augmented z-axis fluctuation of flowing platelets caused by the physical presence of erythrocytes is speculated to be the cause of the hematocrit-dependent increase in platelet adhesion.

Automatic Aortic Valve Cusps Segmentation from CT Images Based on the Cascading Multiple Deep Neural Networks.

Accurate morphological information on aortic valve cusps is critical in treatment planning. Image segmentation is necessary to acquire this information, but manual segmentation is tedious and time consuming. In this paper, we propose a fully automatic aortic valve cusps segmentation method from CT images by combining two deep neural networks, spatial configuration-Net for detecting anatomical landmarks and U-Net for segmentation of aortic valve components. A total of 258 CT volumes of end systolic and end diastolic phases, which include cases with and without severe calcifications, were collected and manually annotated for each aortic valve component. The collected CT volumes were split 6:2:2 for the training, validation and test steps, and our method was evaluated by five-fold cross validation. The segmentation was successful for all CT volumes with 69.26 s as mean processing time. For the segmentation results of the aortic root, the right-coronary cusp, the left-coronary cusp and the non-coronary cusp, mean Dice Coefficient were 0.95, 0.70, 0.69, and 0.67, respectively. There were strong correlations between measurement values automatically calculated based on the annotations and those based on the segmentation results. The results suggest that our method can be used to automatically obtain measurement values for aortic valve morphology.

A computational model study of the influence of the anatomy of the circle of willis on cerebral hyperperfusion following carotid artery surgery.

BACKGROUND: Cerebral hyperperfusion syndrome develops in a small subset of patients following carotid artery surgery (CAS) performed to treat severe carotid artery stenosis. This syndrome has been found to have a close correlation with cerebral hyperperfusion occurring after CAS. The purpose of this study is to investigate whether and how the anatomy of the Circle of Willis (CoW) of the cerebral circulation influences post-CAS cerebral hyperperfusion. METHODS: A computational model of the cerebral circulation coupled with the global cardiovascular system has been developed to investigate hemodynamic events associated with CAS. Nine topological structures of the CoW were investigated in combination with various distribution patterns of stenosis in the feeding arteries of the cerebral circulation. RESULTS: The occurrence of post-CAS cerebral hyperperfusion was predicted for the CoW structures that have poor collateral pathways between the stenosed cerebral feeding arteries and the remaining normal feeding arteries. The risk and the localization of post-CAS hyperperfusion were determined jointly by the anatomy of the CoW and the distribution pattern of stenosis in the cerebral feeding arteries. The presence of basilar artery stenosis or contralateral ICA stenosis increased the risk of post-CAS hyperperfusion and enlarged the cerebral region affected by hyperperfusion. For a certain CoW structure, the diameters of the cerebral communicating arteries and the severity of carotid artery stenosis both had a significant influence on the computed post-CAS cerebral hyperperfusion rates. Moreover, post-CAS cerebral hyperperfusion was predicted to be accompanied with an excessively high capillary transmural pressure. CONCLUSIONS: This study demonstrated the importance of considering the anatomy of the CoW in assessing the risk of post-CAS cerebral hyperperfusion. Particularly, since the anatomy of the CoW and the distribution pattern of stenosis in the cerebral feeding arteries jointly determine the risk and localization of post-CAS cerebral hyperperfusion, a patient-specific hemodynamic analysis aimed to help physicians identify patients at high risk of cerebral hyperperfusion should account for the combined effect of the anatomy of cerebral arteries and cerebral feeding artery stenoses on cerebral hemodynamics.

Biomechanical Gain in Joint Excursion from the Curvature of the Achilles Tendon: Role of the Geometrical Arrangement of Inflection Point, Center of Rotation, and Calcaneus.

The dorsal movement of the Achilles tendon during ankle rotation is restricted by anatomical obstructions. Previously, we demonstrated that the anatomical obstruction provides a gain (gainAT) in the proximal displacement of the calcaneus compared to the change in the Achilles tendon length. Here, we empirically validate and extend our previous modeling study by investigating the effects of a broad range of obstruction locations on gainAT. The largest gainAT could be achieved when the obstruction was located on the most ventral and distal sides within the physiological range of the Achilles tendon, irrespective of the ankle position.

Numerical study on sector-vortex phased irradiation method using annular array transducer in High-Intensity Focused Ultrasound treatment.

Sector-vortex phased irradiation from annular array transducer was numerically studied with breast model constructed from MRI data of real patient. Phase compensation (PC) based on time reversal pre-computation was applied in order to handle phase delay caused by heterogeneity of breast tissues, and results showed great effectiveness on single-focus case, insignificant effectiveness on multi-focus cases with 4 and 8 phase-sectors, but ineffectiveness on multi-focus case with 12 phase-sectors, where enormous undesired outer ablation occurred. For single-focus case, phase compensation not only produced real focus very close to targeted site (0.1 mm deviation), but also decreased thermal peak ratio (outer/focal) largely by 30%. However, phase compensation did not increase total ablated size. For multi-focus cases with 4 and 8 phase-sectors, deformed focal shapes by tissue heterogeneity were restored by phase compensation, but the 4-phase-sector case had higher thermal peak ratio and smaller ablation than 8-phase-sector case for strong cancelling effect between phase-sector borders. Ineffectiveness of phase compensation on multi-focus case with 12 phase-sectors had three considerable reasons. 1st, inequality of piezo-element number between sectors; 2nd, heterogeneous attenuation of breast model; 3rd, insufficient number of piezo-elements per sector; where the 2nd reason originated from breast model, and other two reasons were related to array transducer. This research gave several preliminary indications. 1st, ineffectiveness of phase compensation occurs on case with large phase-sector number when using annular array transducer; 2nd, with same input energy and same irradiation time, sector-vortex phased irradiation creates smaller focal ablation, but withstands longer than single-focus irradiation free of outer ablation; 3rd, phase-difference π between neighboring phase-sectors is disadvantageous because of energy loss; 4th, phase compensation is effective on single-focus for improving pinpoint ablation but not for increasing total ablated size.

A penalized spline fitting method to optimize geometric parameters of arterial centerlines extracted from medical images.

In order to grasp the spatial and temporal evolution of vascular geometry, three-dimensional (3D) arterial bending structure and geometrical changes of arteries and stent grafts (SG) must be quantified using geometrical parameters such as curvature and torsion along the vasculature centerlines extracted from medical images. Here, we develop a robust method for constructing smooth centerlines based on a spline fitting method (SFM) such that the optimized geometric parameters of curvature and torsion can be obtained independently of digitization noise in the images. Conventional SFM consists of the 3rd degree spline basis function and 2nd derivative penalty term. In contrast, the present SFM uses the 5th degree spline basis function and 3rd and 4th derivative penalty terms, the coefficients of which are derived by the Akaike information criterion. The results show that the developed SFM can reduce the errors of curvature and torsion compared to conventional SFM. We then apply the present SFM to the centerline of the SG in an abdominal aortic aneurysm (AAA), and those of bilateral internal carotid arteries (ICA) in 6 cases: 3 cases with aneurysms and 3 cases without any aneurysm. The SG centerlines were obtained from temporal medical images at three scan times. The strong peak of the curvature could be clearly observed in the distal area of the SG, the inversion of the torsion at 0 months in the middle area of SG disappeared over time, and the torsions around the SG bifurcation at the three time periods were inverted. The curvature-torsion graphs along the ICA centerlines superimposing five aneurysmal positions were useful for investigating the relationship between arterial bending structure and aneurysmal positions. Both ICAs had curvature peak values higher than 0.4 within the ICA syphons. The ICA torsion graphs indicated that left and right ICA tended to be a right- and left-handed helix, respectively. In the left ICA syphon, the biggest aneurysm could be observed downstream of the salient torsion inversion. All aneurysms for 3 cases were positioned at the downstream of the inverted torsion.

Characterization of Brain-Targeted Drug Delivery Enhanced by a Combination of Lipid-Based Microbubbles and Non-Focused Ultrasound.

The combination of focused ultrasound (FUS) and microbubbles, an ultrasound (US) contrast agent, has attracted much attention for its ability to open the blood brain barrier (BBB) and deliver drugs to the brain parenchyma. FUS can concentrate US energy in a restricted space, whereas non-focused US can affect a wide area of tissue. Non-focused US is also promising for drug delivery to the brain and other tissues. We have previously developed lipid-based microbubbles (LBs), and demonstrated that non-focused US and LBs have potential for drug delivery to tumor tissues. In this study, to achieve efficient and safe brain-targeted drug delivery, we evaluated the characteristics of BBB opening using non-focused US and LBs. Our results indicated that LBs could induce BBB opening with non-focused US. US frequency and intensity affected the efficiency of BBB opening and brain damage, and showed that the dose of LBs was also related to the efficiency of BBB opening. Furthermore, the combination of non-focused US and LBs could deliver macromolecules at 2000 kDa to the brain, and the induction of BBB opening was found to be reversible. These results suggest that the combination of non-focused US and LBs has potential as a brain-targeted drug delivery system.