Evaluating the Effects of Water Balloons on High-Intensity Focused Ultrasound for Treating Uterine Fibroids.

OBJECTIVE: In the treatment of uterine fibroids with ultrasound-guided high-intensity focused ultrasound (HIFU), water balloons are considered to be a valuable aid for improving safety and efficiency. However, the water balloons worsen the pathway for acoustic transmission, causing degraded performance both in ultrasound therapy and in ultrasound imaging. This study was aimed at establishing a protocol to evaluate the effects of the water balloon. METHODS: Simulations and experiments were carefully conducted to quantitatively investigate the effects of water ballons on the efficiency of HIFU energy delivery and on the quality of ultrasound guiding images. More specifically, HIFU-induced temperature increases in the focal region, together with spatial resolution, contrast and signal-to-noise ratio in the ultrasound guiding images, were compared under the conditions with and without the water balloon. RESULTS: Experiment results revealed that the use of water balloons led to decreases in temperature up to 10ºC within the focal region in some specific situations, but the quality of the guiding images was relatively less affected. CONCLUSION: The study provided knowledge on what influence the water balloon could have in ultrasound-guided HIFU treatment; it also established a practical and standardized evaluation scheme for further optimizing the water balloon, for example, its material and internal liquid compositions. This study can potentially help improve the efficiency and safety of treating uterine fibroids with ultrasound-guided HIFU systems.

Effect of sling exercise therapy on surface electromyography and muscle thickness of superficial cervical muscle groups in female patients with chronic neck pain.

BACKGROUND: The persistence of symptoms in patients with chronic neck pain is considered to be associated with variation in the neck muscle structure and associated neuromuscular control. Sling exercise therapy (SET) has been demonstrated to relieve the symptoms of chronic neck pain, whereas it is controversial whether this benefit is correlated to altered neck muscle structure and associated neuromuscular control in the patients. OBJECTIVE: To investigate the effect of SET on cervical muscle structure (thickness) and associated neuromuscular control in patients with chronic neck pain. METHODS: Twenty-five patients with chronic neck pain were randomly assigned to the SET group (n= 12) or the control group (n= 13). The SET group received the SET intervention for 4 weeks, while the control group maintained normal activities of daily living. At baseline and after 4 weeks of intervention, Visual analogue scale and neck disability index were measured in both groups, and changes in the thickness of the superficial cervical muscles were assessed using musculoskeletal ultrasound. Surface electromyography (EMG) was adapted to assess the neuromuscular control of the neck while the participant was performing the cranio-cervical flexion test. RESULTS: At 4 weeks, the SET group had a significant reduction of RMS in both UT and SCM of EMG compared to the control group (p< 0.05). Regarding ultrasound, the SET group had significantly lower muscle thickness compared to the control group in both the rest position and the MVIC position (p< 0.05). There were no within-group differences in the control group (p> 0.05), while the SET group showed significant reductions in both RMS and muscle thickness (p< 0.05). CONCLUSION: 4-week SET was effective in reducing pain and dysfunction in patients with chronic neck pain, which may be related to improved neck muscle thickness and neuromuscular control of the neck.

Analysis of correlation between heart failure in the early stage of acute myocardial infarction and serum pregnancy associated plasma protein-A, prealbumin, C-reactive protein, and brain natriuretic peptide levels.

BACKGROUND: The study sought to analyze the predictive value of the early measurement of pregnancy associated plasma protein-A (PAPP-A), prealbumin (PAB), C-reactive protein (CRP), brain natriuretic peptide (BNP), and other indicators of heart failure (HF) after acute myocardial infarction (AMI). METHODS: A total of 200 AMI patients admitted to Wuyi People's Hospital from November 1, 2019 to October 31, 2020 were continuously enrolled as the research objects and divided into a HF group (Killip class II or above, n=94) and HF-free group (Killip class I or below, n=106) according to the Killip Classification for Heart Function. RESULTS: The age, creatine kinase-myocardial band (CK-MB), PAPP-A, CRP, suppression of tumorigenicity 2 (ST2), BNP, aldosterone (ALD), and left ventricular end-diastolic diameter (LVEDD) of the HF group were all significantly higher than those of the HF-free group, while the PAB and left ventricular ejection fraction (LVEF) were significantly lower (P<0.05). The PAPP-A, CRP, and BNP of HF patients increased as the Killip class increased, while the PAB decreased progressively (P<0.05). After including the statistically significant single factors in a multivariate logistic regression analysis, it was found that PAPP-A, PAB, CRP, and BNP were the independent influencing factors causing HF in the early stage of AMI; and the diagnostic efficacy of HF in the early stage of AMI (from high to low) was combined test, PAPP-A, PAB, BNP and CRP. CONCLUSIONS: Serum PAPP-A, PAB, CRP and BNP levels are the independent influencing factors of HF after AMI, but comprehensive tests of clinical indicators, including PAPP-A, PAB, CRP, and BNP, are more accurate at predicting and evaluating HF and can be used to guide clinical decisions.

Inhibition of miR-218-5p reduces myocardial ischemia-reperfusion injury in a Sprague-Dawley rat model by reducing oxidative stress and inflammation through MEF2C/NF-κB pathway.

Following myocardial ischemia, myocardial reperfusion injury causes oxidative stress (OS) and inflammation, leading to myocardial cell apoptosis and necrosis. Recently, emerging studies have shown that microRNAs (miRNAs) contribute to the pathophysiology associated with myocardial ischemia-reperfusion (I/R). In this study, we conducted both in-vitro and in-vivo experiments to explore the role of miR-218-5p in ischemia-reperfusion (I/R)- or oxygen and glucose deprivation/reperfusion (OGD/R)-mediated cardiomyocyte injury. A total 44 Sprague-Dawley (SD) rats were used, and randomly divided into four groups, control group (n = 11), miR-218-5p-in group (n = 11), I/R group (n = 11), I/R + miR-218-5p-in group (n = 11). Our data showed that miR-218-5p was overexpressed in H9C2 cardiomyocytes under OGD/R treatment. miR-218-5p inhibition reduced the lactate dehydrogenase (LDH) activity and the levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD), as well as the expression of tumor necrosis factor alpha (TNF-α), interleukin (IL-1ß), and IL-6. Oppositely, miR-218-5p overexpression aggravated OGD/R-mediated damage on H9C2 cells, whereas nuclear factor kappa B (NF-κB) pathway inhibition or myocyte enhancer factor 2C (MEF2C) upregulation reversed miR-218-5p mimics-mediated effects. Bioinformatics analysis predicted that miR-218-5p targeted and dampened its expression, which was testified by the dual-luciferase reporter assay and RNA pull-down assay. In vivo, inhibiting miR-218-5p declined LDH activities and ROS, MDA and SOD levels in rat myocardial tissues under I/R injury, alleviated myocardial fibrosis and inflammatory reactions, and reduced myocardial infarction area. Overall, inhibition of miR-218-5p choked oxidative stress and inflammation in myocardial I/R injury via targeting MEF2C/NF-κB axis, thus relieving the disease progression.

Development and optimization of doxorubicin loaded poly(lactic-co-glycolic acid) nanobubbles for drug delivery into HeLa cells.

Microbubbles (MBs, usually 2-8 microm) as ultrasound contrast agent and drug carrier are promising for ultrasonic imaging and drug delivery. However, MBs posed some limitations due to their large diameters. In the current study, we developed a nanoscale bubbles (nanobubbles, NBs) by encapsulating the doxorubicin (DOX) into poly(lactic-co-glycolic acid) (PLGA) shells (denoted as DOX-PLGA NBs) for drug delivery into cancer cells. The size, morphology, particle stability, drug encapsulation efficiency, and drug payload were determined. The results showed that the DOX-PLGA NBs were uniform (270 +/- 3 nm) and spherical with a smooth surface, and were well dispersed and stable in water. The encapsulation efficiency and payload of DOX increased with its initial loading concentrations. The release behavior of DOX from the DOX-PLGA NBs exhibited a biphasic pattern characterized by an initial burst release followed by a slower and continuous release at both pH 7.4 and pH 4.4, and also presented in a pH-triggered releasing profile. The qualitative analysis of cellular internalization into HeLa cells by inverted fluorescence microscope showed that the cellular uptake of DOX-PLGA NBs was both concentration- and time-dependent. Moreover, the cell viability was also investigated using CCK-8 assay. It was found that DOX-PLGA NBs showed greater HeLa cell growth inhibition effect in vitro compared with free DOX. It was concluded that the DOX-PLGA NBs were biocompatible and appropriate for anti-cancer drug delivery, and were potentially promising as a new therapeutic system for cancer treatment.

Notch-1 signaling promotes the malignant features of human breast cancer through NF-κB activation.

The aberrant activation of Notch-1 signaling pathway has been proven to be associated with the development and progression of cancers. However, the specific roles and the underlying mechanisms of Notch-1 signaling pathway on the malignant behaviors of breast cancer are poorly understood. In this study, using multiple cellular and molecular approaches, we demonstrated that activation of Notch-1 signaling pathway promoted the malignant behaviors of MDA-MB-231 cells such as increased cell proliferation, colony formation, adhesion, migration, and invasion, and inhibited apoptosis; whereas deactivation of this signaling pathway led to the reversal of the aforementioned malignant cellular behaviors. Furthermore, we found that activation of Notch-1 signaling pathway triggered the activation of NF-κB signaling pathway and up-regulated the expression of NF-κB target genes including MMP-2/-9, VEGF, Survivin, Bcl-xL, and Cyclin D1. These results suggest that Notch-1 signaling pathway play important roles in promoting the malignant phenotype of breast cancer, which may be mediated partly through the activation of NF-κB signaling pathway. Our results further suggest that targeting Notch-1 signaling pathway may become a newer approach to halt the progression of breast cancer.

Roles for GP IIb/IIIa and αvß3 integrins in MDA-MB-231 cell invasion and shear flow-induced cancer cell mechanotransduction.

Adhesion of cancer cell to endothelial cells and the subsequent trans-endothelial migration are key steps in hematogenous metastasis. However, the molecular mechanisms of cancer cell/endothelial cell interaction under hemodynamic shear flow and how shear flow-induced cancer cell mechanotransduction are yet to be fully defined. In this study, we identified that the integrins of both platelet glycoprotein IIb/IIIa (GP IIb/IIIa) and αvß3 were crucial for hematogenous metastasis of human breast carcinoma MDA-MB-231 cells. The cell migration and invasion were studied by using Millicell cell culture insert system. The numbers of invaded MDA-MB-231 cells significantly increased by thrombin-activated platelets and reduced by eptifibatide, a platelet inhibitor. Meanwhile, RGDWE peptides, a specific inhibitor of αvß3 integrin, also inhibited MDA-MB-231 cell invasion. We further used a parallel-plate flow chamber to investigate MDA-MB-231 cell adhesion under flow conditions. Alike in static condition, the adhesion capability of MDA-MB-231 cells to endothelial monolayer was also significantly affected by GP IIb/IIIa and αvß3 integrins. The expression of matrix metalloproteinase-2 (MMP-2), MMP-9 and αvß3 integrin in MDA-MB-231 cells were up-regulated after low shear stress exposure (1.84 dynes/cm(2), 2 h). Moreover, we also demonstrated that low shear stress induced a sustained activation of p85 (a regulatory subunit of PI3K) and Akt. Pre-treating MDA-MB-231 cells with the specific PI3K inhibitor of LY294002 abolished the shear stress induced-Akt activation, and the expression of MMP-2, MMP-9, vascular endothelial growth factor (VEGF) and αvß3 integrin were also down-regulated. Immunofluorescence assay showed that low shear stress also induced αvß3 integrin clustering and nuclear factor-κB (NF-κB) activation. Interestingly, shear stress-induced activation of Akt and NF-κB was attenuated by LM609, a specific antibody of αvß3 integrin. It suggests that αvß3 integrin might be as a mechanosensor to trigger both PI3K/Akt and NF-κB signaling pathways. Taken together, these results establish that GP IIb/IIIa and αvß3 integrins are essential mediators, and provide insight into how shear stress-induced αvß3 integrin activation and the downstream pathways for contribution to MDA-MB-231 cell adhesion, migration and invasion.

VCAM-1-targeted core/shell nanoparticles for selective adhesion and delivery to endothelial cells with lipopolysaccharide-induced inflammation under shear flow and cellular magnetic resonance imaging in vitro.

Multifunctional nanomaterials with unique magnetic and luminescent properties have broad potential in biological applications. Because of the overexpression of vascular cell adhesion molecule-1 (VCAM-1) receptors in inflammatory endothelial cells as compared with normal endothelial cells, an anti-VCAM-1 monoclonal antibody can be used as a targeting ligand. Herein we describe the development of multifunctional core-shell Fe(3)O(4)@SiO2 nanoparticles with the ability to target inflammatory endothelial cells via VCAM-1, magnetism, and fluorescence imaging, with efficient magnetic resonance imaging contrast characteristics. Superparamagnetic iron oxide and fluorescein isothiocyanate (FITC) were loaded successfully inside the nanoparticle core and the silica shell, respectively, creating VCAM-1-targeted Fe(3)O(4)@SiO2(FITC) nanoparticles that were characterized by scanning electron microscopy, transmission electron microscopy, fluorescence spectrometry, zeta potential assay, and fluorescence microscopy. The VCAM-1-targeted Fe(3)O(4)@SiO2(FITC) nanoparticles typically had a diameter of 355 ± 37 nm, showed superparamagnetic behavior at room temperature, and cumulative and targeted adhesion to an inflammatory subline of human umbilical vein endothelial cells (HUVEC-CS) activated by lipopolysaccharide. Further, our data show that adhesion of VCAM-1-targeted Fe(3)O(4)@SiO2(FITC) nanoparticles to inflammatory HUVEC-CS depended on both shear stress and duration of exposure to stress. Analysis of internalization into HUVEC-CS showed that the efficiency of delivery of VCAM-1-targeted Fe(3)O(4)@SiO2(FITC) nanoparticles was also significantly greater than that of nontargeted Fe(3)O(4)@SiO2(FITC)-NH2 nanoparticles. Magnetic resonance images showed that the superparamagnetic iron oxide cores of the VCAM-1-targeted Fe(3)O(4)@SiO2(FITC) nanoparticles could also act as a contrast agent for magnetic resonance imaging. Taken together, the cumulative adhesion and uptake potential of these VCAM-1-targeted Fe(3)O(4)@SiO2(FITC) nanoparticles targeted to inflammatory endothelial cells could be used in the transfer of therapeutic drugs/genes into these cells or for diagnosis of vascular disease at the molecular and cellular levels in the future.

Polymeric microbubbles for ultrasonic molecular imaging and targeted therapeutics.

Gas-filled microbubbles ultrasound agent have received wide attention, not only because they can improve ultrasound signals, but also they can be used as drug/gene carriers. Among all types of microbubbles fabricated by different membrane materials and core gases, polymer-shell microbubbles are highly promising. Polymeric microbubbles are more stable than other soft shell microbubbles in vivo. Under destructive ultrasound, polymer-stabilized microbubbles disintegrate and emit a strong non-linear signal, which enables ultrasound imaging with superior sensitivity. Except for ultrasound imaging, polymeric microbubbles could also be applied as drug/gene-delivery system. The thick polymeric shells allow loading a large amount of drugs. Meanwhile, site-specific targeting and controlled drug release in the area of interest can be realized through chemical and physical modification. In this review, we highlight some of the recent examples on polymeric microbubbles and their applications in ultrasound molecular imaging and drug delivery.

The roles of platelet GPIIb/IIIa and alphavbeta3 integrins during HeLa cells adhesion, migration, and invasion to monolayer endothelium under static and dynamic shear flow.

During their passage through the circulatory system, tumor cells undergo extensive interactions with various host cells including endothelial cells and platelets. Mechanisms mediating tumor cell adhesion, migration, and metastasis to vessel wall under flow condition are largely unknown. The aim of this study was to investigate the potential roles of GPIIb/IIIa and alphavbeta3 integrins underlying the HeLa-endothelium interaction in static and dynamic flow conditions. HeLa cell migration and invasion were studied by using Millicell cell culture insert system. The numbers of transmigrated or invaded HeLa cells significantly increased by thrombin-activated platelets and reduced by eptifibatide, a platelet inhibitor. Meanwhile, RGDWE peptides, a specific inhibitor of alphavbeta3 integrin, also inhibited HeLa cell transmigration. Interestingly, the presence of endothelial cells had significant effect on HeLa cell migration regardless of static or cocultured flow condition. The adhesion capability of HeLa cells to endothelial monolayer was also significantly affected by GPIIb/IIIa and alphavbeta3 integrins. The arrested HeLa cells increased nearly 5-fold in the presence of thrombin-activated platelets at shear stress condition (1.84 dyn/cm(2) exposure for 1 hour) than the control (static). Our findings showed that GPIIb/IIIa and alphavbeta3 integrins are important mediators in the pathology of cervical cancer and provide a molecular basis for the future therapy, and the efficient antitumor benefit should target multiple receptors on tumor cells and platelets.