Microbubbles carrying resiquimod enhance HIFU ablation and co-stimulate tumor immune response / 中华超声影像学杂志

Objective:To prepare the resiquimod-loaded lipid microbubbles R848-MBs, evaluate their enhanced ultrasound imaging and high intensity focused ultrasound (HIFU) ablation effects, and explore their ability to improve tumor immune microenvironment synergize with HIFU.Methods:R848-MBs were prepared by the thin film hydration-mechanical shock method; The basic characteristics and safety of R848-MBs were detected, the HIFU controlled-release characteristics were verified in vitro and the drug metabolism and biological distribution were investigated in vivo. The ability of enhancing ultrasound imaging was observed in vitro and in vivo. To investigate the enhanced HIFU ablation effect of R848-MBs, six EMT6 tumor-bearing mice were randomly divided into HIFU group and R848-MBs+ HIFU group, three mice in each group, the changes in contrast average sound intensity before and after ablation in mouse tumor areas and the change of ultrasound image gray value in tumor area were evaluated, the tumor were resected to observe the coagulative necresis by TTC staining and HE staining. Forty-five tumor-bearing mice were randomly divided into control group, Free R848 group, HIFU group, Blank-MBs+ HIFU group and R848-MBs+ HIFU group, nine mice in each group. On the third day after treatment, 3 mice in each group were randomly selected and killed, to evaluate the ability of R848-MBs to improve tumor immune microenvironment synergize with HIFU. The expression level of CRT on the surface of tumor cells were detected by immunofluorescence staining, the proportion of mature DC in lymph nodes, spleen, and CD8 + T cells in spleen were detected by flow cytometry. The treatment effectiveness of each group( n=6) were evaluated by measuring tumor volume, observing and drawing survival curves. Results:The R848-MBs lipid microbubbles with good safety were successfully prepared, with a concentration of 2.58×10 9/ml, as spherical bubbles under optical microscope and laser confocal microscopy, in a particle size of (1.72±0.11)μm, at a surface potential of (-10.16±0.73)mV. The cumulative drug release was up to 83.44% after HIFU (90 W, 3 s) in vitro. The concentration of R848 in plasma decreased rapidly, and the drug concentration in tumor tissue of the R848-MBs+ HIFU group was higher than that of the R848 group 24 hours after treatment ( P<0.01). The ultrasound imaging of R848-MBs was significantly enhanced in contrast mode in vitro and in vivo; R848-MBs can significantly enhance the HIFU ablation effect, the contrast average sound intensity change in the tumor area before and after ablation in the R848MBs+ HIFU group was greater than that in the R848 group ( P<0.05), and the immediate ultrasound grayscale value change in the HIFU+ R848-MBs group was 46.34±3.21, which was significantly greater than that in the HIFU group (10.67±1.53), with statistical significance ( P<0.000 1). Coagulation necrosis was observed in tumor HE staining and TTC staining. The results of treatment efficacy in vivo showed that R848-MBs+ HIFU group had the strongest therapeutic effect, and R848-MBs combined with HIFU treatment could significantly prolong the survival period of mice compared with intravenous injection of free R848 ( P<0.01). Immunofluorescence staining and flow cytometry results showed an increase in the expression level of CRT on the surface of tumor tissue in the R848-MBs combined with HIFU group, and the percentage of mature DC in tumor draining lymph nodes (58.53±1.04)% were significantly higher than those in the HIFU group (37.56±2.13)% ( P<0.001), and the percentage of mature DC in the spleen (70.65±1.91)% were significantly higher than those in the HIFU group (36.46±3.89)% ( P<0.001), the percentage of CD8 + T cells in the spleen (27.46±3.04)% was significantly higher than that in the HIFU group (18.69±0.29)% ( P<0.01). Conclusions:The HIFU controlled-release lipid microbubbles R848-MBs can not only enhance the efficiency of HIFU ablation, but also improve the tumor immune microenvironment.

Effect of ultrasonic microbubbles combined with gentamicin in the treatment of diabetic foot ulcer under low-frequency ultrasound / 中华超声影像学杂志

Objective:To investigate the effect of microbubbles combined with gentamicin on the clearance of bacterial biofilms and the healing of diabetic foot ulcers under low-frequency ultrasound.Methods:From July 2021 to June 2022, 27 patients with chronic diabetic foot ulcers complicated with infection were prospectively selected from the Trauma Center of the Second Affiliated Hospital of Chongqing Medical University. The patients were divided into low-frequency ultrasound + microbubbles + gentamicin ointment group, low-frequency ultrasound + microbubbles group, and gentamicin ointment group by using a random number table, with 9 patients in each group. The three groups were all treated with simple debridement by the same surgeon.Afterward, in the low-frequency ultrasound+ microbubbles+ gentamicin ointment group, the wounds covered by 4% microbubble suspension were firstly irradiated with low-intensity focused ultrasound for 5 min, and then evenly applied with gentamicin ointment. In the low-frequency ultrasound + microbubbles group, the wounds covered by 4% microbubble suspension were irradiated with low-intensity focused ultrasound for 5 min. The gentamicin ointment group was treated with gentamicin ointment evenly. The treatment lasted for 2 weeks, and secretions and tissue specimens were collected during and 2 weeks after the treatment, respectively. The general indexes of wound surface (including ulcer depth score, secretion exudation score, fresh granulation tissue growth score, and total index score), ulcer area, ulcer healing rate, as well as negative rate of secretion culture were compared among the three groups after treatment. Additionally, the structural changes in bacterial biofilms under a scanning electron microscope and colony count under a laser confocal scanning microscope were compared among the three groups after treatment.Results:No significant differences were found in the general datas among the three groups (all P>0.05). After treatment for 2 weeks, the overall general indexes showed statistically and significant differences among the three groups (all P<0.05). Each index score in the low-frequency ultrasound + microbubbles + gentamicin ointment group was lower than that in the low-frequency ultrasound + microbubbles group and the gentamicin ointment group (all P<0.05). There were no significant differences in overall ulcer area among the three groups ( P>0.05). The overall ulcer healing rate presented significant differences among the three groups ( P<0.05). The healing rate in the low-frequency ultrasound + microbubbles + gentamicin ointment group was higher than that in the low-frequency ultrasound + microbubbles group and the gentamicin ointment group (all P<0.05). The overall negative rates of secretion culture among the three groups were significantly different ( P<0.05), the negative rate in the low-frequency ultrasound + microbubbles + gentamicin ointment group was higher than that in the low-frequency ultrasound + microbubbles group and the gentamicin ointment group (all P<0.05). Scanning electron microscopy confirmed bacterial biofilm infection in the three groups before treatment. After treatment for 2 weeks, the biofilm formation in the low-frequency ultrasound + microbubbles + gentamicin ointment group reduced significantly, while the low-frequency ultrasound + microbubbles group and the gentamicin ointment group had little change compared with that before treatment. Significant differences were detected in total colony count among the three groups under the confocal microscope ( P<0.05). The colony count in the low-frequency ultrasound + microbubbles + gentamicin ointment group was lower than that in the low-frequency ultrasound + microbubbles group and the gentamicin ointment group (both P<0.05). Conclusions:Ultrasound microbubbles combined with gentamicin can clear bacterial biofilms and promote the healing of diabetic foot ulcers.

A Sonotrombólise Promove Melhora dos Índices de Motilidade e Perfusão do Ventrículo Esquerdo após o Infarto Agudo do Miocárdio / Sonothrombolysis Promotes Improvement in Left Ventricular Wall Motion and Perfusion Scores after Acute Myocardial Infarction

Resumo Fundamento Demonstrou-se recentemente que a aplicação de ultrassom de alta energia com microbolhas, técnica conhecida como sonotrombólise, causa a dissolução de trombos intravasculares e aumenta a taxa de recanalização angiográfica no infarto agudo do miocárdio com supradesnivelamento do segmento ST (IAM-CSST). Objetivo Avaliar o efeito da sonotrombólise nos índices de motilidade e perfusão miocárdicas em pacientes com IAM-CSST, utilizando a ecocardiografia com perfusão miocárdica em tempo real (EPMTR). Método Uma centena de pacientes com IAM-CSST foram randomizados em dois grupos: Terapia (50 pacientes tratados com sonotrombólise e angioplastia coronária primária) e Controle (50 pacientes tratados com angioplastia coronária primária). Os pacientes realizaram EPMTR para analisar a fração de ejeção do ventrículo esquerdo (FEVE), o índice de escore de motilidade segmentar (IEMS) e o número de segmentos com defeito de perfusão miocárdica, 72 horas após o IAM-CSST e com 6 meses de acompanhamento. Foi considerado significativo p < 0,05. Resultados Pacientes tratados com sonotrombólise apresentaram FEVE mais alta que o grupo Controle em 72 horas (50 ± 10% vs. 44 ± 10%; p = 0,006), e essa melhora foi mantida em seis meses (53 ± 10% vs. 48 ± 12%; p = 0,008). O IEMS foi similar nos grupos Terapia e Controle em 72 horas (1,62 ± 0,39 vs. 1,75 ± 0,40; p = 0,09), mas tornou-se menor no grupo Terapia em 6 meses (1,46 ± 0,36 vs. 1,64 ± 0,44; p = 0,02). O número de segmentos com defeito de perfusão não foi diferente entre os grupos em 72 horas (5,92 ± 3,47 vs. 6,94 ± 3,39; p = 0,15), mas ficou menor no grupo Terapia em 6 meses (4,64 ± 3,31 vs. 6,57 ± 4,29; p = 0,01). Conclusão A sonotrombólise em pacientes com IAM-CSST resulta na melhora dos índices de motilidade e perfusão ventricular ao longo do tempo.

Abstract Background It has recently been demonstrated that the application of high-energy ultrasound and microbubbles, in a technique known as sonothrombolysis, dissolves intravascular thrombi and increases the angiographic recanalization rate in patients with ST-segment-elevation myocardial infarction (STEMI). Objective To evaluate the effects of sonothrombolysis on left ventricular wall motion and myocardial perfusion in patients with STEMI, using real-time myocardial perfusion echocardiography (RTMPE). Methods One hundred patients with STEMI were randomized into the following 2 groups: therapy (50 patients treated with sonothrombolysis and primary coronary angioplasty) and control (50 patients treated with primary coronary angioplasty). The patients underwent RTMPE for analysis of left ventricular ejection fraction (LVEF), wall motion score index (WMSI), and number of segments with myocardial perfusion defects 72 hours after STEMI and at 6 months of follow-up. P < 0.05 was considered statistically significant. Results Patients treated with sonothrombolysis had higher LVEF than the control group at 72 hours (50% ± 10% versus 44% ± 10%; p = 0.006), and this difference was maintained at 6 months of follow-up (53% ± 10% versus 48% ± 12%; p = 0.008). The WMSI was similar in the therapy and control groups at 72 hours (1.62 ± 0.39 versus 1.75 ± 0.40; p = 0.09), but it was lower in the therapy group at 6 months (1.46 ± 0.36 versus 1.64 ± 0.44; p = 0.02). The number of segments with perfusion defects on RTMPE was similar in therapy and control group at 72 hours (5.92 ± 3.47 versus 6.94 ± 3.39; p = 0.15), but it was lower in the therapy group at 6 months (4.64 ± 3.31 versus 6.57 ± 4.29; p = 0.01). Conclusion Sonothrombolysis in patients with STEMI resulted in improved wall motion and ventricular perfusion scores over time.

Preparation method and characteristics of biomimetic microbubbles / 中华超声影像学杂志

Objective:To evaluate the feasibility and applicability of using phospholipid-hybridization method for preparing biomimetic microbubbles (Bio-MBs) ultrasound contrast agents.Methods:Leukocyte biomimetic microbubbles (MB leu), platelet biomimetic microbubbles (MB pla) and erythrocyte biomimetic microbubbles (MB ery) were prepared by multiple steps: film-hydration, phospholipid-hybridization, mechanical oscillation. The size and zeta potential of Bio-MBs were measured by dynamic light scattering. A laser scanning confocal microscopy experiment was performed to confirm the presence of membrane proteins on the shell of Bio-MBs. The fluorescence of FITC-labeled typical membrane protein was evaluated using a flow cytometer. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to characterize the membrane protein. Biosafety of Bio-MBs was evaluated by CCK-8 counting kit, blood and major organs. The contrast enhancement effect and stability were observed in vitro and in vivo. An in vivo fluorescence imaging system was performed to evaluate the distribution of Bio-MBs. The application value of biomimetic microbubbles was measured by ultrasound molecular imaging by using ischemia-reperfusion rat models and acute hepatitis rat models. Results:Bio-MBs with spherical shape distributed homogenously, without obvious aggregation. The membrane proteins were successfully integrated into the shell of Bio-MBs.The diameter of three Bio-MBs was similar to that of control microbubbles (MB con) ( P>0.05), three Bio-MBs had a lower zeta potential than MB con ( P<0.05). The Bio-MBs had an appreciable performance in vitro and in vivo biosafety. The Bio-MBs retained the main proteins inherited from cell membrane. Contrast enhanced ultrasound imaging in vitro and in vivo showed that the Bio-MBs had a stable imaging ability.MB leu and MB pla have good targeted imaging effect in two disease models. Conclusions:A series of Bio-MBs ultrasound contrast agents, which have high stability, biosafety and targeted imaging efficiency, were successfully prepared by using phospholipid-hybridization method. This fabrication method for obtaining Bio-MBs can be applied to different clinical scenarios with different cell types in the future.

Experimental study of dual-targeted microbubbles in assessing the expression levels of pro-angiogenic markers during the growth of renal carcinoma / 中华超声影像学杂志

Objective:To explore the capability of vascular endothelial growth factor receptor 2 (VEGFR2)/integrinα vβ 3 dual-targeted microbubbles in assessing the expression level of pro-angiogenic factors during renal cell carcinoma (RCC) growth. Methods:VEGFR2/integrinα vβ 3 dual-targeted microbubbles were prepared by using biotin-avidin linkage method. Twenty subcutaneous RCC xenografts in nude mice were established by subcutaneously injecting 786-O cells and then divided into 2 groups randomly. The targeted contrast-enhanced ultrasound (t-CEUS) examination was performed for all 10 mice in the first group when xenograft tumors were metered from 5 to 10 mm and >10 to 20 mm respectively. And the quantitative parameters of RCC on t-CEUS were longitudinally evaluated during tumor growth. The second group were divided into two subgroups according to xenograft tumors′ diameter, which was 5 to 10 mm and >10 to 20 mm respectively, and underwent t-CEUS examination. Quantitative analysis was performed for all t-CEUS images to obtain the targeted quantitative parameters, which including peak intensity (PI), area under the time-intensity curve (AUC), the differential tissue enhancement (dTE, presenting the difference in PI before (P 1) and after (P 2) the process of Flash). All xenograft tumors in the second group were harvested for immunohistochemical staining to observe the expression of VEGFR2, integrinα vβ 3 and CD31, and their differences in RCC with different tumor sizes. And the correlations between quantitative parameters and VEGFR2, integrinα vβ 3 and CD31 were analyzed. Results:The longitudinal comparison showed that there were statistically significant differences between AUC and dTE of RCC with different tumor sizes (all P<0.001). The larger the tumor size, the smaller the parameters were. According to the horizontal comparison, the expression levels of VEGFR2 and integrinα vβ 3 in larger RCCs were higher than those of RCCs with smaller size (both P<0.05), but there was no significant difference in CD31 expression between the two subgroups ( P=0.754). Both the targeted quantitative parameters (AUC anddTE ) and pro-angiogenic factors (VEGFR2 and integrinα vβ 3) were negatively correlated with tumor size ( rs=-0.83, -0.81, -0.70, -0.88; all P<0.05). Further more, there were good positive correlations between AUC and VEGFR2, integrinαvβ ( rs=0.76, 0.72; all P<0.05). There were good positive correlations between dTE and VEGFR2, integrinα vβ 3 ( rs=0.81, 0.70; all P<0.05). Additionally, the parameter PI was positively correlated with the expression of CD31 ( rs=0.70, P=0.025). Conclusions:The t-CEUS, mediated by VEGFR/integrinα vβ 3 dual-targeted microbubbles, allows noninvasive assessment of the expression levels of VEGFR2 and integrinα vβ 3 in RCCs, which decrease gradually with the increase of tumor size.

Clarificação de águas de lavagem de filtros em estações de tratamento de água por coagulação, floculação e flotação por ar dissolvido / Clarification of spent filter backwash water in water treatment plants by coagulation, flocculation and dissolved air flotation

RESUMO Este artigo apresenta estudos de coagulação, floculação e flotação por ar dissolvido que foram conduzidos com o objetivo de promover a clarificação de águas de lavagens de filtros (ALF) em uma estação de tratamento de água (ETA) com base em experimentos de bancada e piloto. A efetividade da coagulação-floculação foi analisada pela flotabilidade dos agregados obtidos sob diferentes níveis de pH e dosagem de cloreto de polialumínio, enquanto ensaios de flotação foram investigados com base na eficiência de saturação em bancada e na taxa de recirculação. Os resultados indicaram que o pH e a concentração do coagulante representaram condições críticas na etapa de coagulação. A variação do pH da ALF condicionada com 8 mg.L-1 de Al+3 destacou a formação de espécies hidrolisáveis e a elevada atividade das cargas dos coloides indicou a ocorrência de mecanismos diferentes da neutralização de cargas na coagulação. A máxima clarificação da água ocorreu com dosagem de 12 mg.L-1 de Al+3, razão de recirculação de 10%, tempo de floculação de 15 min e pH 6,5. Em configuração otimizada, a técnica alcançou remoção de 88% de turbidez e 86% de sólidos totais, o que pode resultar em mais de 11 quilogramas de sólidos removidos do sistema de tratamento em cada ciclo de lavagem em comparação com a sedimentação, implicando aumento de 33% da carreira de filtração e, consequentemente, redução de custos e melhor desempenho da estação.

ABSTRACT This article presents studies on coagulation, flocculation and dissolved air flotation that were conducted with the aim of promoting the clarification of spent filter backwash water (SFBW) in a Water Treatment Plant (WTP) based on bench and pilot experiments. The effectiveness of coagulation-flocculation was analyzed according to the buoyancy of the aggregates obtained from different levels of pH and dosage of polyaluminium chloride, whereas flotation tests were investigated based on bench saturation efficiency and recirculation rate. According to the results, the pH and the concentration of the coagulant represented critical conditions in the coagulation. The pH variation of the SFBW conditioned with 8 mg.L-1 of Al + 3 highlighted the formation of hydrolyzable species, and the high activity of the colloid indicated the occurrence of different mechanisms for neutralization in the coagulation. The maximum clarification of the SFBW occurred with a dosage of 12 mg.L-1 of Al+3, recirculation rate of 10%, flocculation time of 15 min, and pH 6.5. In an optimized configuration, the technique achieved removal of 88% turbidity and 86% total solids, which can result in more than 11 kilograms of solids removed from the WTP in each backwashing cycle compared with sedimentation. This would imply an increase of 33% in the backwashing interval and, consequently, in the reduction of costs and in the better performance of the WTP.

Use of contrast-enhanced ultrasound in pediatrics / Utilização da ultrassonografia com contraste de microbolhas na pediatria

Abstract Although contrast-enhanced ultrasound has been shown to provide considerable benefits, particularly in pediatric patients, it is still used relatively rarely in Brazil. It has proven to be a safe technique, and adverse effects are rare. In this review, we address the technique and main applications of contrast-enhanced ultrasound in the pediatric population, including the evaluation of focal liver lesions, abdominal trauma, kidney grafts, liver grafts, bowel loops, and vesicoureteral reflux. It is important for pediatric radiologists to be acquainted with this promising tool, understanding its applications and limitations.

Resumo A ultrassonografia com contraste de microbolhas é um método ainda pouco utilizado no Brasil, mas que tem grandes vantagens, ainda mais na faixa pediátrica, como ausência de radiação e dispensa de sedação/anestesia. O contraste de microbolhas tem se mostrado bastante seguro e com raríssimas reações adversas graves. Apresentaremos nesta revisão a técnica para o uso desse contraste na ultrassonografia, bem como as principais aplicações na faixa pediátrica, como avaliação das lesões hepáticas focais, do trauma abdominal, do enxerto hepático e renal nos transplantes, das alças intestinais e da pesquisa de refluxo vesicoureteral. É importante que radiologistas que exerçam funções em centros pediátricos conheçam essa ferramenta promissora.

Applicability of B-mode ultrasonography, ARFI elastography and contrast-enhanced ultrasound in the evaluation of chronic kidney disease in dogs / Aplicabilidade da ultrassonografia modo-B, elastografia ARFI e ultrassonografia contrastada na avaliação da doença renal crônica em cães

The objective of this study was to verify the applicability of B-mode ultrasonography, ARFI elastography and CEUS in the diagnosis of chronic kidney disease and its Stages in dogs. 24 healthy dogs and 28 with CKD were included. In B-mode, the echogenicity, echotexture and cortico-medullary ratio of the kidneys were verified. By elastography, the shear-wave velocity of the cortical (SWVcort) and medullary (SWVmed) regions were determined and tissue deformity was evaluated. Wash-in, wash-out and peak enhancement (TPic) of the contrast in the renal parenchyma were calculated and homogeneity, presence of filling gaps and distinction of filling phases were evaluated by CEUS. Changes in echogenicity, echotexture and cortico-medullary ratio were observed only in sick patients. There was an increase in SWVcort in CKD, with a cutoff point >2.91m/s. Healthy kidneys were non-deformable and 25% had changes in gray scales. There was an increase in wash-in and TPic, changes in filling characteristics, filling failures and difficulty in distinguishing between the Stages in CEUS in CKD. It was found that dogs with CKD 2, 3 and 4 had greater SWVcort and wash-in values than CKD 1. Elastographic and CEUS changes were observed in dogs with CKD, demonstrating the applicability of ultrasonographic techniques in their diagnosis.(AU)

O objetivo deste estudo foi verificar a aplicabilidade da ultrassonografia modo-B, elastografia ARFI e CEUS no diagnóstico da doença renal crônica e seus estágios em cães. Foram incluídos 24 cães saudáveis e 28 com DRC. Pelo modo-B, verificou-se ecogenicidade, ecotextura e relação córtico-medular dos rins. Pela elastografia, foram determinadas as velocidades de cisalhamento das regiões cortical (SWVcort) e medular (SWVmed) e foi avaliada a deformidade tecidual. Calculou-se os temos de wash-in, wash-out e pico de intensidade (TPic) do parênquima renal e avaliou-se homogeneidade, presença de falhas e distinção das fases de preenchimento por contraste por meio de CEUS. Alterações em ecogenicidade, ecotextura e relação córtico-medular foram observadas somente em pacientes doentes. Houve aumento da SWVcort na DRC, com ponto de corte >2.91m/s. Rins saudáveis foram não-deformáveis e 25% apresentou alterações nas escalas de cinza. Houve aumento do wash-in e TPic, alterações nas características de preenchimento, falhas e dificuldade para distinguir as fases de preenchimento do contraste nos rins com DRC. Verificou-se que cães com DRC graus 2, 3 e 4 tiveram valores de SWVcort e wash-in maiores que cães com DRC grau 1. Foram obsrervadas alterações elastográfica e de CEUS em cães com DRC, demonstrando a aplicabilidade destas técnicas ultrassonográficas no seu diagnóstico.(AU)

Applicability of B-mode ultrasonography, ARFI elastography and contrast-enhanced ultrasound in the evaluation of chronic kidney disease in dogs

ABSTRACT: The objective of this study was to verify the applicability of B-mode ultrasonography, ARFI elastography and CEUS in the diagnosis of chronic kidney disease and its Stages in dogs. 24 healthy dogs and 28 with CKD were included. In B-mode, the echogenicity, echotexture and cortico-medullary ratio of the kidneys were verified. By elastography, the shear-wave velocity of the cortical (SWVcort) and medullary (SWVmed) regions were determined and tissue deformity was evaluated. Wash-in, wash-out and peak enhancement (TPic) of the contrast in the renal parenchyma were calculated and homogeneity, presence of filling gaps and distinction of filling phases were evaluated by CEUS. Changes in echogenicity, echotexture and cortico-medullary ratio were observed only in sick patients. There was an increase in SWVcort in CKD, with a cutoff point >2.91m/s. Healthy kidneys were non-deformable and 25% had changes in gray scales. There was an increase in wash-in and TPic, changes in filling characteristics, filling failures and difficulty in distinguishing between the Stages in CEUS in CKD. It was found that dogs with CKD 2, 3 and 4 had greater SWVcort and wash-in values than CKD 1. Elastographic and CEUS changes were observed in dogs with CKD, demonstrating the applicability of ultrasonographic techniques in their diagnosis.

RESUMO: O objetivo deste estudo foi verificar a aplicabilidade da ultrassonografia modo-B, elastografia ARFI e CEUS no diagnóstico da doença renal crônica e seus estágios em cães. Foram incluídos 24 cães saudáveis e 28 com DRC. Pelo modo-B, verificou-se ecogenicidade, ecotextura e relação córtico-medular dos rins. Pela elastografia, foram determinadas as velocidades de cisalhamento das regiões cortical (SWVcort) e medular (SWVmed) e foi avaliada a deformidade tecidual. Calculou-se os temos de wash-in, wash-out e pico de intensidade (TPic) do parênquima renal e avaliou-se homogeneidade, presença de falhas e distinção das fases de preenchimento por contraste por meio de CEUS. Alterações em ecogenicidade, ecotextura e relação córtico-medular foram observadas somente em pacientes doentes. Houve aumento da SWVcort na DRC, com ponto de corte >2.91m/s. Rins saudáveis foram não-deformáveis e 25% apresentou alterações nas escalas de cinza. Houve aumento do wash-in e TPic, alterações nas características de preenchimento, falhas e dificuldade para distinguir as fases de preenchimento do contraste nos rins com DRC. Verificou-se que cães com DRC graus 2, 3 e 4 tiveram valores de SWVcort e wash-in maiores que cães com DRC grau 1. Foram obsrervadas alterações elastográfica e de CEUS em cães com DRC, demonstrando a aplicabilidade destas técnicas ultrassonográficas no seu diagnóstico.

Experiência inicial com ultrassom Doppler com contraste por microbolhas em adição ao ultrassom Doppler convencional para seguimento de correção endovascular de aneurisma de aorta abdominal / Initial experience with Duplex scan combined with contrast-enhanced ultrasound for follow-up of endovascular abdominal aortic aneurysm repair

Resumo Contexto O ultrassom contrastado por microbolhas (CMUS) é uma modalidade diagnóstica de acurácia bem demonstrada por estudos internacionais para seguimento de reparo endovascular do aneurisma de aorta abdominal (EVAR). Não existem, no entanto, estudos nacionais focados nesse método de seguimento. Objetivos O objetivo deste estudo foi relatar a experiência inicial com CMUS em um hospital terciário, traçando uma comparação dos achados do CMUS com o ultrassom Doppler convencional (USGD), com o intuito de verificar se a adição de contraste ao protocolo padrão de controle ultrassonográfico incorreu alteração nos achados. Métodos Entre 2015 e 2017, 21 pacientes em seguimento de EVAR foram submetidos ao USGD seguido de CMUS. Foram avaliados os achados de exame referentes à identificação de complicações, bem como à capacidade de identificação da origem da endofuga. Resultados Entre os 21 casos avaliados, 10 complicações foram evidenciadas no total: sete pacientes apresentaram endofuga (33,3%); dois pacientes apresentaram estenose em ramo de endoprótese (9,52%); e um paciente apresentou dissecção em artéria ilíaca externa (4,76%). Em 21 pacientes avaliados, o uso combinado dos métodos identificou 10 casos de complicações pós-EVAR. Em seis dos sete casos de endofugas (85,71%), o uso dos métodos combinados foi capaz de identificar a origem. O USGD isolado falhou na identificação da endofuga em dois casos (28,5%), identificando achados duvidosos em outros dois casos (28,5%), que obtiveram definição diagnóstica após associação do CMUS. Conclusões O CMUS é uma técnica de fácil execução, a qual adiciona subsídios ao seguimento de EVAR infrarrenal.

Abstract Background Microbubble contrast enhanced ultrasound (CEUS) is an accurate diagnostic method for follow-up after endovascular abdominal aortic aneurysm repair (EVAR) that has been well-established in international studies. However, there are no Brazilian studies that focus on this follow-up method. Objectives The objective of this study was to report initial experience with CEUS at a tertiary hospital, comparing the findings of CEUS with those of conventional Doppler ultrasound (DUS), with the aim of determining whether addition of contrast to the standard ultrasonographic control protocol resulted in different findings. Methods From 2015 to 2017, 21 patients in follow-up after EVAR underwent DUS followed by CEUS. The findings of these examinations were analyzed in terms of identification of complications and their capacity to identify the origin of endoleaks. Results There was evidence of complications in 10 of the 21 cases examined: seven patients exhibited endoleaks (33.3%); two patients exhibited stenosis of a branch of the endograft (9.52%); and one patient exhibited a dissection involving the external iliac artery (4.76%). In the 21 patients assessed, combined use of both methods identified 10 cases of post-EVAR complications. In six of the seven cases of endoleaks (85.71%), use of the methods in combination was capable of identifying the origin of endoleakage. DUS alone failed to identify endoleaks in two cases (28.5%) and identified doubtful findings in another two cases (28.5%), in which diagnostic definition was achieved after employing CEUS. Conclusions CEUS is a technique that is easy to perform and provides additional support for follow-up of infrarenal EVAR.

Research progress of focused ultrasound combined with microbubbles to open blood-brain barrier for brain-targeted delivery / 药学学报

Blood-brain barrier (BBB) is the most challengeable obstacle for brain-targeted drug delivery. The combination of focused ultrasound (FUS) and microbubbles provides a new way for brain-targeted drug delivery across BBB based on the cavitation effect. This review briefly described the recent research of FUS combined with microbubbles to enhance the BBB permeability for brain-targeted drug delivery. The contents included the FUS mechanism, the types of the commercial microbubbles, shell materials, inner gas and preparation techniques of microbubbles, the opening mechanism of FUS with microbubbles, and the safety consideration. FUS combined with microbubbles may be the effective strategy to improve the BBB permeability for brain-targeted delivery, which could provide references for the clinical applications.

Targeted contrast enhanced ultrasound for in vivo quantitative evaluation on angiogenesis in transplanted ovarian of rats / 中国医学影像技术

Objective: To observe the value of targeted contrast enhanced ultrasound (CEUS) using anti-Mullerian canal hormone (AMH) targeted nanobubbles (AMH-NB) for in vivo quantitative evaluation on ovarian neovascular density after ovarian auto-transplantation in SD rats. Methods: The nanobubbles carrying anti-AMH antibody were prepared, and their basic physical properties were examined. Then ovarian auto-transplantation rat models were established. The targeted (AMH-NB), non-targeted (N-NB) contrast agents and SonoVue were administered on the 7th day after transplantation to obtain peak intensity (PI) and time to peak (TTP). The microvascular density (MVD) was measured using immunohistochemistry, and the correlation of PI, TTP and MVD were analyzed. Results: The particle size of AMH-NB uniformly distributed, ranged (622.67±33.65)nm, and the concentration of AMH-NB was (2.90±0.26)×108/ml. PI of ovarian angiography with AMH-NB was (7.93±0.65)dB and TTP was (42.53±1.74)s, with N-NB was (6.14±0.44)dB and (54.35±1.73)s, with Sonovue was (4.15±0.83)dB and (28.71±1.18)s, respectively (all P<0.05).Immunohistochemical analysis showed that the microvascular density was (61.20±6.84)/HP, histological analysis indicated that AMH-NB were able to penetrate blood vessels to the interstitial space and combine with AMH. PI, TTP of AMH-NB were highly both correlated with MVD (r=0.84, r=-0.84, both P<0.05). Conclusion: AMH-NB can be used to qualitatively and quantitatively evaluate the angiogenesis in transplanted ovarian of rats in vivo with CEUS.

Ultrasound application in brain disease treatment:research progress / 国际药学研究杂志

Because of the blood-brain barrier (BBB)and the lack of significant enhancement method for the drug penetration into and retention in brain, effective methods and drugs for the treatment of brain diseases are extremely limited. Ultrasound can effec tively, safely and reversibly open the BBB and thus is an ideal method for the targeted, non-invasive drug entry into the intracranial tis sue, which has been expected to become a new means for treating brain diseases in the near future. Following a brief introduction of some biological characteristics of brain, this review elaborates the mechanisms of the ultrasound combining microbubbles(USMB)for brain targeted drug delivery, including the opening of the tight junction, stimulation of endocytosis, inhibition of multidrug resistance protein expression, enhancement of drug concentration in target area, extension of the residence time, and adjustment of the intersti tial pressure in target tissue. Then, the application of ultrasound in the treatment of some diseases including the Alzheimer's disease (AD), Huntington's disease(HD), Parkinson's disease(PD), and brain tumors are summarized, and the existing problems in the ul trasound therapy are also discussed, in order to provide a valuable reference for further improvement in the treatment of brain diseases.

Assessment of neovascularization during bone healing using contrast-enhanced ultrasonography in a canine tibial osteotomy model: a preliminary study

Impact of diagnostic ultrasound with different mechanical index enhanced by microbubbles on chemotherapy of nude mice bearing pancreatic tumor / 中国介入影像与治疗学

Objective: To investigate the impact of diagnostic ultrasound (DUS) with different mechanical index (MI) combined with microbubbles on tumor blood flow and chemotherapeutic drug concentration in nude mice bearing human pancreatic cancer. Methods: Pancreatic tumor models were established through injecting tumor cell suspension into bilateral hind limbs in 33 nude mice,which were divided into three groups (group A, B and C) randomly. Contrast-enhanced ultrasound (CEUS), chemotherapeutic agents injection and exposure/sham exposure with DUS were performed on all models in the three groups. For each model, one of the hind limb was chosen as therapeutic side (exposure), and the other hind limb as control side (sham exposure). CEUS was performed before and after ultrasonic exposure/sham to obtain the time-intensity curve (TIC), peak intensity (PI) and AUC. Doxorubicin was injected through the tail vein before ultrasonic exposure/sham exposure. And microbubbles were continuously injected during ultrasonic exposure/sham exposure. MI of therapeutic side for ultrasonic exposure in group A, B and C was 0.3, 0.7 and 1.1, respectively. After ultrasonic exposure/sham exposure, Doxorubicin concentration in vitro tumor tissue was detected, and then histological analysis was done. In addition, the range of peak negative pressure correspond with different MI were detected before ultrasonic exposure/sham exposure. Results: Concentration of Doxorubicin of therapeutic side in group A was obviously higher than that of control side ([1.45±0.53]μg/g vs [1.07±0.46]μg/g; t=-5.163, P=0.001). In group B and C, there was no statistically significant difference of Doxorubicin concentration between therapeutic side and control side in the drug concentration (Z=-0.297, -0.357, P=0.766, 0.721). Quantitative analysis of CEUS showed that both PI and AUC increased obviously after ultrasonic exposure/sham exposure compared with those before exposure/sham exposure (all P<0.05), except for PI of control side in group B and AUC of control side in group B and C. Neither hemorrhage nor swelling was observed in tumor tissue of both sides in any group. The range of peak negative pressure correspond with MI of 0.3 (group A), 0.7 (group B) and 1.1 (group C) was 0.81-0.83 MPa, 0.96-1.32 MPa and 2.29-2.53 MPa, respectively. Conclusion: DUS (MI=0.3) combined with microbubbles can enhance blood perfusion and improve the chemotherapeutic drug concentration of tumor tissues in nude mice bearing human pancreatic cancer.

Research progress of microbubble ultrasound contrast agent for treatment of uterine fibroids with high intensity focused ultrasound / 中国介入影像与治疗学

As a non-invasive thermal ablation technique, high intensity focused ultrasound (HIFU) has been widely used in treatment of uterine fibroids with affirmed efficacy. Microbubble contrast agent has important clinical value in the visualization of lesions, the evaluation of efficacy after HIFU ablation and the mechanism of synergy. The advancements of microbubble contrast agent in treatment of uterine fibroids with HIFU were reviewed in this article.

Ultrasound-mediated sPD-1 and miR-206 gene nanoscale microbubbles synergistic inhibition of H22 hepatoma subcutaneous xenografts in mice / 中国医学影像技术

Objective: To investigate the effect of ultrasound-mediated soluble programmed cell death 1 receptor (sPD-1) and miR-206 loaded nanoscale microbubbles on H22 hepatoma subcutaneous xenografts in mice. Methods: sPD-1 and miR-206 loaded nanoscale microbubbles were prepared. The mice models of H22 hepatoma xenografts were established and randomly divided into model group, microbubble control group, miR-206 microbubble group, sPD-1 microbubble group and combined group (miR-206 and sPD-1 microbubbles). Mice in each group (each n=8) were treated with normal saline or corresponding nanoscale microbubbles every 2 days by injection via tail vein, and then irradiated by ultrasound once after every injection. Tumor tissues were obtained after being treated 5 times. Tumor volume and quality were measured, the volume and quality inhibitory rates were calculated. HE staining was used to observe pathological changes of the tumors. The expressions of Bcl-2, Bax proteins were detected by immunohistochemistry. The expressions of Bcl-2, Bax, c-met, interferon-γ (IFN-γ) and programmed cell death 1 receptor ligand (PD-L1) mRNA were detected with RT-PCR. Quantitative real-time fluorescence PCR was used to detect the expression of miR-206.Results: The nanoscale bubbles were spherical and distributed uniformly. Compared with model group, tumor volume and quality decreased in other groups, and the volume and quality inhibitory rates increased (all P<0.05), especially in combined group (all P<0.05). Compared with model group, the Bax protein and mRNA expressions both increased, whereas the Bcl-2 protein and mRNA expressions decreased in other groups, especially in combined group (all P<0.01). There were significant differences of Bax, Bcl-2, c-met, PD-L1, IFN-γ and miR-206 mRNA in tumor tissues among each group (all P<0.01). Conclusion: Ultrasound-mediated sPD-1 combine miR-206 loaded nanoscale microbubbles can synergistically inhibit H22 hepatoma xenografts in mice.

Effects of ultrasound combined with microbubbles on intracellular Ca 2+ homeostasis in carboplatin-treated microenvironment of A549 cell line / 中华超声影像学杂志

Objective To investigate the effects of ultrasound combined with microbubbles on intracellular Ca2+ homeostasis in carboplatin ( CBP )-treated A549 cell and its possible mechanisms of inhibiting A549 cell line activity . Methods According to whether SonoVue was used or not ,and the different dose of CBP ,the groups A-F were arranged as the ultrasound(US) group(group A) ,the ultrasound combined with microbubbles ( USMB) group( group B) ,the low dose CBP ( 100 μg/ml) + US group( group C) ,the low dose CBP+USMB group( group D) ,the high dose CBP ( 200 μg/ml)+ US group ( group E) and the high dose CBP+USMB group( group F) .A549 cells were bathed and washed by a calcium-free buffer , loaded with Ca2+ indicator fluo-4 AM . Real-time images were acquired using laser confocal microscopy .The fluorescence intensity of intracellular calcium ion concentration ([Ca2+ ]i) in individual living cell was observed and the calcium overload was analyzed . Results After ultrasound irradiation ,the normalized fluorescence intensity of [Ca2+ ]i increased rapidly ,then returned to a new homeostasis (selected cells in groups A ,B ,E ,F) or experienced a second calcium oscillation ( some cells in group C and D) . All the selected cells in group B and some cells in group C and D exhibited superimposed oscillations . The calcium overloading time in group D was longer than those of any other groups . Four cells in group A experienced delayed calcium oscillations . Compared with group A ,the selected cells in other groups exhibited a larger amplitude of calcium oscillation( all P < 0 .05) and the selected cells in group B and D exhibited calcium oscillation for a longer period of time( all P <0 .05) . Conclusions In the calcium-free buffer ,US ,USMB , CBP+ US ,CBP + USMB are direct stimuli of calcium overload in A 549 cells . SonoVue ,CBP ,CBP +SonoVue are all synergistic stimuli of calcium overload in A 549 cells irradiated by ultrasound .US ,USMB and CBP may synergistically induce calcium release from intracellular store sites in A 549 cells . Calcium overload is a possible mechanism of ultrasound combined with microbubbles in assisting CBP chemotherapy .

Experimental study of IL-8 monoclonal antibody combined with ultrasound targeted microbubble destruction on myocardial ischemia/reperfusion injury in rabbits / 中华超声影像学杂志

Objective To explore the application value of IL-8 monoclonal antibody microbubble combined with ultrsound targeted microbubble destruction ( UTMD) on alleviating myocardial ischemia reperfusion/injury ( MIRI) in rabbits .Methods Forty-two rabbits were randomly divided into closed chest group ( n =7) ,open chest control group ( n = 7) and ischemia-reperfusion ( I/R) group ( n = 28) .I/R group were randomly divided into 30 min reperfusion group( n =7) ,60 min reperfusion group( n =7) ,120 min reperfusion group ( n = 7 ) and 180 min reperfusion group ( n = 7 ) .All rabbits were examined by electrocardiogram , echocardiography and HE staining after MIRI . Targeted myocardial contrast echocardiography ( MCE) was performed and ELISA was used to detect IL-8 content in rabbit myocardium before and after UTMD . Results Electrocardiogram and wall motion returned to normal at 60 min after reperfusion .Targeted MCE showed that with the prolongation of reperfusion after I/R ,the video intensity of myocardium in reperfusion area increased gradually , reaching its peak at 120 min and 180 min after reperfusion .After UTMD ,the video intensity decreased ,and the change rate of video intensity in 30 min reperfusion group was higher than those in other reperfusion groups(all P<0 .05) .The content of IL-8 and its neutralization rate in the ELISA results were consistent with the video intensity and rate of change of targeted MCE .HE staining and scanning electron microscopy showed that myocardial injury was found in I/R group .With the prolongation of reperfusion time ,the degree of myocardial injury was gradually aggravated ,and the injury was alleviated after irradiation .Conclusions IL-8 monoclonal antibody combined with UTMD has the advantages of non-invasive and highly effective in alleviating MIRI .It provides a new way to treat MIRI .

H2 O2/PFP-loaded acoustic phase-change lipid nanoparticles enhancing thrombolysis in artificial vessel for simulation of coronary circulation:an in-vitro study by low intensity focused ultrasound irradiation / 中华超声影像学杂志

Objective To prepare a novel H2 O2-loaded and phase-change nanoparticles ,combined with low intensity focused ultrasound (LIFU) to dissolve intravascular thrombosis for the evaluation of its value in the treatment of coronary artery thrombolysis in the patients with acute myocardial infarction ( AMI) . Methods H2 O2-loaded and phase-change nanoparticles were prepared by membrane hydration method . 50 ml arterial blood was collected from rabbit carotid artery and made into 400-800 mg thrombus blocks . They were divided into four groups randomly :poly butylene succinate buffer ( PBS) solution as blank control (group A) ,SonoVue microbubbles group(group B) ,PFP phase-change nanoparticles group ( group C ) , H2 O2-loaded and phase-change nanoparticles group ( group D ) . Each group of thrombus samples were placed into the artificial blood vessels of the extracorporeal thrombolysis simulation cycle device and combined with LIFU irradiation . The weights and pathological presentations were compared before and after thrombolysis . The volume of O2 in the vascular system was inspected using Dissolved Oxygen Detector ( DOD) after LIFU exposure . Results The average size of H2 O2-loaded nano-droplets was ( 456 .7 ± 31 .2) nm . Fluorescence microscopy showed that the structure of nanoparticles was stable and uniform in size . The weight loss and rates of thrombus in the four groups after thrombolysis were:group A (52.2±11.5)mg,(9.7±3.5)％ ;groupB(110.0±21.9)mg,(19.4±2.4)％ ;groupC(239.6±46.3)mg, (39 .7 ± 6 .3)％ and group D (309 .8 ± 44 .8)mg ,(54 .2 ± 6 .6)％ ,respectively . The weight loss and rates of thrombus in group C and D were higher than those in group A and B ( all P < 0 .01) ,but there was no significant difference between group C and D ( P >0 .05) . The DOD showed that along with the increase of H2O2 concentration ,O2 release was rising . Conclusions H2O2/PFP nanoparticles with LIFU performs efficient thrombolysis and provides O2 to improve hypoxia ,which provides a potential novel treatment method for AMI patients with coronary embolism .