Targeted delivery of MerTK protein via cell membrane engineered nanoparticle enhances efferocytosis and attenuates atherosclerosis in diabetic ApoE-/- Mice.

BACKGROUND: Clearance of apoptotic cells by efferocytosis is crucial for prevention of atherosclerosis progress, and impaired efferocytosis contributes to the aggravated atherosclerosis. RESULTS: In this study, we found that diabetic ApoE-/- mice showed aggravated atherosclerosis as hyperglycemia damaged the efferocytosis capacity at least partially due to decreased expression of Mer tyrosine kinase (MerTK) on macrophages. To locally restore MerTK in the macrophages in the plaque, hybrid membrane nanovesicles (HMNVs) were thus developed. Briefly, cell membrane from MerTK overexpressing RAW264.7 cell and transferrin receptor (TfR) overexpressing HEK293T cell were mixed with DOPE polymers to produce nanovesicles designated as HMNVs. HMNVs could fuse with the recipient cell membrane and thus increased MerTK in diabetic macrophages, which in turn restored the efferocytosis capacity. Upon intravenous administration into diabetic ApoE-/- mice, superparamagnetic iron oxide nanoparticles (SMN) decorated HMNVs accumulated at the aorta site significantly under magnetic navigation, where the recipient macrophages cleared the apoptotic cells efficiently and thus decreased the inflammation. CONCLUSIONS:  Our study indicates that MerTK decrease in macrophages contributes to the aggravated atherosclerosis in diabetic ApoE-/- mice and regional restoration of MerTK in macrophages of the plaque via HMNVs could be a promising therapeutic approach.

Supplementation of Clostridium butyricum Alleviates Vascular Inflammation in Diabetic Mice.

Background: Gut microbiota is closely related to the occurrence and development of diabetes and affects the prognosis of diabetic complications, and the underlying mechanisms are only partially understood. We aimed to explore the possible link between the gut microbiota and vascular inflammation of diabetic mice. Methods: The db/db diabetic and wild-type (WT) mice were used in this study. We profiled gut microbiota and examined the and vascular function in both db/db group and WT group. Gut microbiota was analyzed by 16s rRNA sequencing. Vascular function was examined by ultrasonographic hemodynamics and histological staining. Clostridium butyricum (CB) was orally administered to diabetic mice by intragastric gavage every 2 days for 2 consecutive months. Reactive oxygen species (ROS) and expression of nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were detected by fluorescence microscopy. The mRNA expression of inflammatory cytokines was tested by quantitative polymerase chain reaction. Results: Compared with WT mice, CB abundance was significantly decreased in the gut of db/db mice, together with compromised vascular function and activated inflammation in the arterial tissue. Meanwhile, ROS in the vascular tissue of db/db mice was also significantly increased. Oral administration of CB restored the protective microbiota, and protected the vascular function in the db/db mice via activating the Nrf2/HO-1 pathway. Conclusion: This study identified the potential link between decreased CB abundance in gut microbiota and vascular inflammation in diabetes. Therapeutic delivery of CB by gut transplantation alleviates the vascular lesions of diabetes mellitus by activating the Nrf2/HO-1 pathway.

The Combined Effect of Nanobubble-IR783-HPPH-Affibody Complex and Laser on HER2-Positive Breast Cancer.

Introduction: Nanobubble is an innovative ultrasound contrast agent that triggers the development of targeted imaging of HER2-positive breast cancer by combining with HER2 affibody and IR783. HPPH is a second-generation photosensitiser that is effective in treating tumours. Hence, the nanobubble-IR783-HPPH-affibody (NIHA) complex demonstrates considerable potential in the treatment of HER2-positive breast cancer. Methods: We fabricated the NIHA complex via an advanced thin-film hydration method and detected its characteristics such as particle size, morphology, stability, and cytotoxicity. Moreover, the effect of NIHA complex with laser on HER2-positive breast cancer was confirmed via in vitro and in vivo experiments. Results: The NIHA complex was spheroid, stable and exhibited no cytotoxicity; moreover, its particle size was 524.8 ± 53.3 nm (n = 5). In combination with laser treatment, NIHA complex reduced the cell viability and tumour volume, induced apoptosis of HER2-positive breast cancer cells, and prolonged survival of nude mice. Conclusion: The newly prepared NIHA complex with laser treatment has the potential on treating HER2-positive breast cancer.

AHNAK-modified microbubbles for the intracranial delivery of triptolide: In-vitro and in-vivo investigations.

Autophagic dysfunction related cascade events might induce the accumulation of α-synuclein (αSyn) in Parkinson's disease (PD). Triptolide (T10) has been reported as a potential autophagy inducer but limited by hepatotoxicity, low solubility and rapid metabolism. In this study, a novel AHNAK-targeted microbubbles integrated with T10 (T10-AHNAK-MBs) was developed to alleviate motor deficit in rAAV2/5-wild type and A53T mutant αSyn transfected PD mouse model. AHNAK facilitated the accumulation of microbubbles (MBs) near the cerebral vessel wall. Furthermore, bubble cavitation caused by focused-ultrasound (FUS) exposure could simultaneously induce drug release and blood-brain-barrier opening in the area of interest. The results of western blotting, thioflavin S staining, immunofluorescence, ELISA and behavior test demonstrated that T10-AHNAK-MBs with FUS exposure (T10-AHNAK-MBs-FUS) could significantly delivery more T10 into substantia nigra, promote clearance of various forms of αSyn, reduce tyrosine hydroxylase positive neuron loss, restore dopamine secretion, and eventually alleviate motor deficits, along with largely reduced adverse effects. The analyses of autophagic markers suggested that autophagy lysosome pathway (ALP) might dominate the T10-induced αSyn degeneration, including the oligomers and pre-formed fibrils. Thus, T10-AHNAK-MBs-FUS constitutes a promising strategy against the motor deficits in PD by promoting clearance of pathogenic αSyn aggregates via inducing ALP.

Early Assessment of Atherosclerotic Lesions and Vulnerable Plaques in vivo by Targeting Apoptotic Macrophages with AV Nanobubbles.

Background: The early detection of atherosclerotic lesions is particularly important for risk prediction of acute cardiovascular events. Macrophages apoptosis was significantly associated with the degree of AS lesions and especially contributed to plaque vulnerability. In this research, we mainly sought to explore the feasibility of a home-made AV-nanobubbles (NBAV) for visualization of apoptotic macrophages and assessment of atherosclerosis (AS) lesions by contrast-enhanced ultrasound (CEUS) imaging. Methods: NBAV were prepared by "Optimized Thin-Film Hydration" and "Biotin-Avidin-Biotin" methods. Then, the characterization and echogenicity of NBAV were measured and analyzed in vitro. The targeting ability of NBAV to ox-LDL-induced apoptotic macrophages was observed by laser scanning confocal microscope. The ApoE-/- mice mode fed with high fat diet were observed by high-frequency ultrasound, microanatomy and oil red O staining. CEUS imaging in vivo was performed on AS plaques with NBAV and NBCtrl injection through the tail vein in turn in ApoE-/- mice. After CEUS imaging, the plaques were confirmed and analyzed by histopathological and immunological assessment. Results: The prepared NBAV had a nano-scale size distribution with a low PDI and a negative zeta potential. Moreover, NBAV showed an excellent stability and exhibited a significantly echogenic signal than saline in vitro. In addition, we found that NBAV could target apoptotic macrophages induced by ox-LDL. Compared with NBCtrl, CEUS imaging of NBAV showed strong and sustained echo enhancement in plaque area of aortic arch in vivo. Further research showed that NBAV sensitive plaques presented more significant pathological changes with several vulnerable plaque features and abundant TUNEL-positive area. Conclusion: NBAV displayed a sensitive indicator to evaluate apoptotic macrophages, indicating a promising CEUS molecular probe for AS lesions and vulnerable plaques identification.

Colon specific delivery of miR-155 inhibitor alleviates estrogen deficiency related phenotype via microbiota remodeling.

Compelling data have indicated menopause-associated increase in cardiovascular disease in women, while the underlying mechanisms remain largely unknown. It is established that changes of intestinal microbiota affect cardiovascular function in the context of metabolic syndrome. We here aimed to explore the possible link between host intestinal function, microbiota, and cardiac function in the ovariectomy (OVX) mouse model. Mice were ovariectomized to induce estrogen-related metabolic syndrome and cardiovascular defect. Microbiota was analyzed by 16s rRNA sequencing. miRNA and mRNA candidates expression were tested by qPCR. Cardiac function was examined by echocardiography. Colon specific delivery of miRNA candidates was achieved by oral gavage of Eudragit S100 functionalized microspheres. In comparison with the sham-operated group, OVX mice showed compromised cardiac function, together with activated inflammation in the visceral adipose tissue and heart. Lactobacillus abundance was significantly decreased in the gut of OVX mice. Meanwhile, miR-155 was mostly upregulated in the intestinal epithelium and thus the feces over other candidates, which in turn decreased Lactobacillus abundance in the intestine when endocytosed. Oral delivery of miR-155 antagonist restored the protective microbiota and thus protected the cardiac function in the OVX mice. This study has established a possible regulatory axis of intestinal miRNAs-microbiota-estrogen deficiency related phenotype in the OVX model. Colon specific delivery of therapeutic miRNAs would possibly restore the microbiota toward protective phenotype in the context of metabolic syndrome.

Clinical Value of TCCD for Evaluating the Prognosis of Patients with Severe Traumatic Brain Injury After Large Decompressive Craniectomy: A Retrospective Study.

INTRODUCTION: It is challenging to assess the prognosis of patients with severe traumatic brain injury (sTBI) after large decompressive craniectomy (DC). The aim of this study was to evaluate the clinical value of transcranial color-coded duplex sonography (TCCD) for assessing the prognosis of sTBI patients 6 months after large DC. METHODS: This was a retrospective observational study that consecutively enrolled 84 patients with sTBI who were followed up for prognosis until 6 months after large DC. The primary endpoint was the Glasgow Outcome Score (GOS). According to the GOS, patients were divided into an unfavorable prognosis group (GOS 1-3, n = 47) and a favorable prognosis group (GOS 4-5, n = 37). RESULTS: Significant between-group differences were found in age and hemodynamic parameters (systolic peak blood flow velocity, end-diastolic blood flow velocity, mean blood flow velocity, pulsatility index and resistance index) of the middle cerebral artery detected by TCCD (P < 0.05 for all). Subsequently, ridge regression was used to build a prognostic model for patients with large DC. Based on the cerebral hemodynamic parameters measured by TCCD and age, the mean (± standard deviation) area under the curve of the prognostic model in patients with sTBI after large DC was 0.76 ± 0.22. The sensitivity and specificity were 82.08% and 74.17%, respectively. CONCLUSIONS: The cerebral hemodynamic parameters detected by TCCD, combined with age, may be used to predict the outcomes of patients with sTBI at 6 months after large DC. As a noninvasive method, TCCD has the potential to assess the prognosis of these patients. TRIAL REGISTRATION: ChiCTR: ChiCTR1800019758. Registered 27 November 2018-retrospectively registered ( http://www.chictr.org.cn/index.aspx ).

Preliminary study of carotid variables under ultrasound analysis as predictors for the risk of coronary arterial atherosclerosis.

BACKGROUND: Carotid atherosclerosis by ultrasound scanning can be considered as an ideal window to reflect systemic artery atherosclerosis, which has aroused wide concern for predicting the severity of coronary artery atherosclerosis clinically. Ultrasound radio frequency (RF) data technology has enabled us to evaluate the carotid structure and elastic function precisely, for predicting the severity of coronary artery atherosclerosis. METHODS: Patients with suspected coronary artery disease (CAD) underwent coronary angiography and were assigned to four groups according to whether atherosclerotic plaque was found or not and it caused stenosis. Carotid artery intima-media thickness (IMT) and arterial stiffness were investigated by quality intima-media thickness (QIMT) and quality arterial stiffness (QAS) techniques during ultrasound scanning. Univariable and multivariable modeling were used to investigate correlations of carotid parameters to coronary artery atherosclerosis. Receive operating characteristic (ROC) curves were used to evaluate diagnostic performance of these ultrasound variables. RESULTS: Carotid IMT and stiffness variables pulse wave velocity (PWV), α, ß and compliance coefficient (CC) were statistically different between every two-group's comparisons. IMT correlated with stiffness variables significantly with r = 0.70, 0.77, 0.63, and -0.39, respectively. All variables correlated with the severity of coronary atherosclerosis with the odd ratio (OR) of 1.73, 1.67, 1.19, 1.23, and 0.56 accordingly as IMT, PWV, α, ß and CC were concerned. The AUC of IMT, PWV, α, ß and CC were 0.9257, 0.8910, 0.8016, 0.9383, 0.8581 with correctly classified rate of 88.16%, 83.77%, 78.07%, 86.84%, and 81.58%, respectively. CONCLUSIONS: Carotid artery IMT and stiffness variable PWV, α, ß and CC presented favorable predicting and differentiating values for patients with coronary atherosclerosis of different severity.

The severity of portal hypertension by a non-invasive assessment: acoustic structure quantification analysis of liver parenchyma.

BACKGROUND: Acoustic structure quantification (ASQ) has been applied to evaluate liver histologic changes by analyzing the speckle pattern seen on B-mode ultrasound. We aimed to assess the severity of portal hypertension (PHT) through hepatic ultrasonography. METHODS: Sixty patients diagnosed with PHT and underwent surgical treatment with portosystemic shunts were enrolled. Portal pressure (PP) was measured intraoperatively. Patients were divided into subgroups according to the severity of gastroesophageal varices and Child-Pugh class. Three difference ratio (Cm2) values on ASQ histogram mode were analyzed for their relationships with PP, degree of gastroesophageal varices and Child-Pugh liver function. Thirty healthy volunteers matched with the patients for gender and age were enrolled as controls. Comparisons among groups and correlation of the parameters with PP were analyzed. Area under the receive operating characteristic curve was used to evaluate the predicting value of ASQ parameters. RESULTS: In the patients, the ASQ parameters peak Cm2 (Cm2max), mean Cm2 (Cm2mean) and the highest occurred Cm2 value of the obtained red curve (RmaxCm2) were all greatly increased (P < 0.0001, P < 0.0001, P = 0.027). Multiple comparisons indicated that, regardless of Child-Pugh class and degree of gastroesophageal varices, the patients had significantly increased Cm2max and Cm2mean compared with the controls (all P < 0.0001). No differences among subgroups were observed. Cm2max was significantly statistically correlated with PP (r = 0.3505, P < 0.01), degree of varices (r = 0.4998, P < 0.0001). Youden's index for Cm2max with a cut-off value of 140.3 for predicting the presence of PHT, gastroesophageal varices and liver function equal to or worse than Child-Pugh class B were 0.8, 0.91 and 0.84, respectively. CONCLUSIONS: ASQ analysis of ultrasonographic images may have a role in the evaluation of the severity of PHT by detecting liver histologic changes in the speckle pattern caused by cirrhosis.

Risk Factors for Anthracycline-Induced Cardiotoxicity.

Background: Several cardiovascular risk factors have been suggested to be associated with anthracycline-induced cardiotoxicity, but their quantitative effects have not reached a consensus. Methods: We searched PubMed, EMBASE, and Cochrane Library databases for manuscripts published from inception to February 2021, which reported the results of cardiotoxicity due to anthracycline chemotherapy without trastuzumab. Cardiotoxicity defined by any reduction of left ventricular eject fraction (LVEF) to below 50% or a >10% reduction from baseline was defined as the primary endpoint. Odd ratios (OR) with 95% confidence intervals (CI) were calculated using a random-effects model meta-analysis. Results: A total of 7,488 patients receiving anthracycline chemotherapy without trastuzumab were included, who had at least one risk factor at baseline. Hypertension (OR: 1.99; 95% CI: 1.43-2.76), diabetes mellitus (OR: 1.74; 95% CI: 1.11-2.74), and obesity (OR: 1.72; 95% CI: 1.13-2.61) were associated with increased risk of cardiotoxicity. In addition, the relative reduction of global longitudinal strain (GLS) from baseline after anthracycline treatment could significantly improve the detection ability of cardiotoxicity (28.5%, 95% CI: 22.1-35.8% vs. 16.4%, 95% CI: 13.4-19.9%) compared with LVEF. The early detection rate of anthracycline-induced cardiotoxicity (3 months after chemotherapy) by GLS was 30.2% (95% CI: 24.9-36.1%), which is similar with the overall result of GLS. Conclusions: Hypertension, diabetes mellitus, and obesity are associated with increased risk of anthracycline-induced cardiotoxicity, which indicates that corresponding protective strategies should be used during and after anthracycline treatment. The findings of higher detection rate and better early detection ability for cardiotoxicity than LVEF added new proofs for the advantages of GLS in detection of AIC.

A Novel Methodology for Semi-automatic Measurement of Arterial Stiffness by Doppler Ultrasound: Clinical Feasibility and Reproducibility.

The conventional manual approach to measurement of aortic pulse wave velocity (PWV) by Doppler ultrasonography is time consuming and operator dependent. Here we report a new semi-automated methodology for more efficient and objective measurement of aortic PWV and results of tests of its clinical feasibility and reproducibility. Carotid-femoral pulse wave velocity (cfPWV) was measured in 50 patients with suspected coronary artery disease (aged 59.2 ± 10.0 y, 36 males) by three independent observers, including two experienced sonographers and one cardiologist without ultrasonographic experience. The cfPWV measured by the semi-automatic method (cfPWVA) was compared with reference values obtained by averaging measurements by two experienced sonographers using the conventional standard manual method (cfPWVM). Measurements of cfPWVA were feasible in all 50 patients and exhibited excellent agreement with averaged cfPWVM from the two experienced sonographers, with an intraclass correlation coefficient (ICC) of 0.915 (95% confidence interval: 0.876-0.942). The inexperienced observer-measured cfPWVA did not differ from the cfPWVM measured by the two experienced sonographers (8.04 ± 1.29 vs. 8.14 ± 1.32 m/s, p > 0.05), with a high consistency by ICC of 0.877 (0.793-0.928). Bland-Altman plots further illustrated the good agreement between the two methods and good intra- and inter-observer reproducibility. Time consumption for cfPWV measurement using the new method was significantly less than that for the manual method (122 ± 35 s vs. 455 ± 105 s, p < 0.0001), saving about 73% of the time. This new semi-automatic methodology for aortic PWV measurement not only has an accuracy similar to that of the conventional standard manual method but is also highly feasible and time saving. It may provide a reliable, simple and reproducible approach to arterial stiffness evaluation in clinical settings.

Preparation of multifunctional nanobubbles and their application in bimodal imaging and targeted combination therapy of early pancreatic cancer.

Pancreatic cancer will gradually become the second leading cause of cancer death due to its poor suitability for surgical treatment, frequent recurrence and metastasis, and insensitivity to radiotherapy and chemotherapy. Strategies for precise early detection and effective targeted treatment of pancreatic cancer are urgently needed. Because of its unique advantages, molecular targeted contrast-enhanced ultrasound imaging (CEUI) has generated new opportunities to overcome this challenge. The aim of this study was to explore multifunctional nanobubbles named IR780-NBs-DTX as novel ultrasound contrast agents (UCAs) for dual-mode targeted imaging and photothermal ablation combined with chemotherapy for pancreatic cancer. An optimized "film hydration method" was used to prepare IR780-NBs-DTX in this research. The characteristics and ability of the new UCAs were detected via in vitro, in vivo and ex vivo experiments. The initial dose of 0.15 mg IR-780 iodide/1.0 mg DTX was considered to be the best formula for IR780-NBs-DTX, and the concentration of 6 ×106 bubbles/mL was best for CEUI. The excellent characteristics of IR780-NBs-DTX, including a uniform nanoscale particle size (349.8± 159.1 nm, n= 3), good performance in dual-mode imaging, high stability and reliable biocompatibility, were also proven. In the in vitro cell experiments, IR780-NBs-DTX targeted more pancreatic cancer cells than the control treatments, and the targeting rate was approximately 95.6± 1.7%. Under irradiation with an 808 nm laser, most cells died. Furthermore, the in vivo study demonstrated that IR780-NBs-DTX could precisely detect pancreatic cancer through near infrared fluorescence (NIRF) imaging and CEUI, and the tumor almost disappeared at 18 days after combined treatment. In ex vivo experiments, immunohistochemistry (IHC) and immunofluorescence (IF) showed that the expression of HSP70 increased and that of PCNA decreased, and many apoptotic tumor cells were observed by TUNEL staining in the IR780-NBs-DTX group. The newly prepared IR780-NBs-DTX are novel nanosized UCAs with high efficiency for dual-mode molecular targeted imaging and combined therapy, and they may have future potential applications in the precise detection and effective targeted therapy of small and metastatic lesions in the early stage of pancreatic cancer.

Effective inhibition of miR-330/SHIP1/NF-κB signaling pathway via miR-330 sponge repolarizes microglia differentiation.

Neuroinflammation mediated by microglia has been identified as vital pathogenesis in Parkinson's disease (PD). This study aimed to investigate the role and potential regulatory mechanism of microRNA-330 in the lipopolysaccharide (LPS)-induced chronic neuroinflammatory model. Primary microglia chronic inflammation model and PD animal model were established by LPS treatment. Bulged microRNA-330 sponges containing six microRNA binding sites were constructed and delivered by plasmid or recombinant adeno-associated virus (rAAV2)/5-green fluorescent protein (GFP) vector. The expression levels of microRNA-330 were assessed by a quantitative real-time polymerase chain reaction. Primary microglia polarization was determined by flow cytometry; meanwhile, dopamine and pro-(anti-)inflammatory cytokines were measured by enzyme-linked immunosorbent assay. Expression levels of GFAP, lba1, inducible nitric oxide synthase (iNOS), Arg1, SHIP1, cytoplasmic, and nuclear factor-κB (NF-κB) were analyzed by Western blot. The behavioral deficit was determined by the rotarod test. The expression of microRNA-330 increased in the first 4 days and reached a plateau subsequently after LPS treatment. The sponges-mediated repression effect on M1 polarization was gradually enhanced with time. Treatment of miR-330 sponges increased the SHIP1 and Arg1 expression, and decreased the translocation of NF-κB and iNOS expression, suggesting the repression of inflammation. In the LPS-induced PD mice, administration of rAAV-sponge-GFP suppressed activation of microglia, downregulated proinflammatory cytokines, resumed the secretion of dopamine, rescued the dopaminergic neurons, and alleviated motor dysfunction. Our results demonstrated that microRNA-330 sponges could sustainably suppress LPS-induced polarization of microglia both in vivo and in vitro probably by negatively regulating NF-κB activity via target SHIP1 in microglia, which might be a promising neuroprotective strategy in neurological diseases, such as PD.

Substantia Nigra Hyperechogenicity Reflects the Progression of Dopaminergic Neurodegeneration in 6-OHDA Rat Model of Parkinson's Disease.

Parkinson's disease (PD) is the second most common neurodegenerative disease, and there is still no effective way to stop its progress. Therefore, early detection is crucial for the prevention and the treatment of Parkinson's disease. The current diagnosis of Parkinson's disease, however, mainly depends on the symptoms, so it is necessary to establish a reliable imaging modality for PD diagnosis and its progression monitoring. Other studies and our previous ones demonstrated that substantia nigra hyperechogenicity (SNH) was detected by transcranial sonography (TCS) in the ventral midbrain of PD patients, and SNH is regarded as a characteristic marker of PD. The present study aimed to explore whether SNH could serve as a reliable imaging modality to monitor the progression of dopaminergic neurodegeneration of PD. The results revealed that the size of SNH was positively related with the degree of dopaminergic neuron death in PD animal models. Furthermore, we revealed that microglia activation contributed to the SNH formation in substantia nigra (SN) in PD models. Taken together, this study suggests that SNH through TCS is a promising imaging modality to monitor the progression of dopaminergic neurodegeneration of PD.

Novel biomimetic dual-mode nanodroplets as ultrasound contrast agents with potential ability of precise detection and photothermal ablation of tumors.

PURPOSE: Molecule-targeted ultrasound imaging has attracted extensive attention for precise diagnosis and targeted therapy of tumors. The aim of this research is to prepare novel biomimetic dual-mode nanoscale ultrasound contrast agents (UCAs), which can not only evade the immune clearance of reticuloendothelial system, but also have the potential ability of precise detection and photothermal ablation of tumors. METHODS: In this study, for the first time, the novel biomimetic UCAs were prepared by encapsulating liquid perfluorohexanes with red blood cell membranes carrying IR-780 iodide and named IR780-RBCM@NDs. The characteristics of that were verified through the particle size analyzer, scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. The stability of IR780-RBCM@NDs at 37 °C was explored. The abilities of immune escape, dual-mode imaging and photothermal effect for IR780-RBCM@NDs were verified via in vitro experiments. RESULTS: The novel prepared nanodroplets have good characteristics such as mean diameter, zeta potential, and relatively stability. Importantly, the integrin-associated protein expressed on the surface of RBCMs was detected on IR780-RBCM@NDs. Then, compared with control groups, IR780-RBCM@NDs performed excellent immune escape function away from macrophages in vitro. Furthermore, the IR-780 iodide was observed on the new nanodroplets and that was able to perform the dual-mode imaging with near-infrared fluorescence imaging and contrast-enhanced ultrasound imaging after the phase change. Finally, the effective photothermal ablation ability of IR780-RBCM@NDs was verified in tumor cells. CONCLUSION: The newly prepared biomimetic IR780-RBCM@NDs provided novel ideas for evading immune clearance, performing precise diagnosis and photothermal ablation of tumor cells.

Brown adipose tissue-derived exosomes mitigate the metabolic syndrome in high fat diet mice.

The ever-increasing incidence of obesity and related disorders impose serious challenges on public health worldwide. Brown adipose tissue (BAT) has strong capacity for promoting energy expenditure and has shown great potential in treating obesity. Exosomes are nanovesicles that share the characteristics of their donor cells. Whether BAT derived exosomes (BAT-Exos) might exert similar metabolic benefits on obesity is worthy of investigation. Methods: Obese mice were established by high-fat-diet (HFD) feeding and were treated with Seum-Exos or BAT-Exos isolated from young healthy mice. Blood glucose, glucose tolerance and blood lipids were tested in mice with indicated treatments. Histology examinations were performed on adipose tissue, liver and heart by HE staining and/or Oil Red O staining. Echocardiography was performed to evaluate cardiac function of mice. In vivo distribution of exosomes was analyzed by fluorescence labeling and imaging and in vitro effects of exosomes were evaluated by cell metabolism analysis. Protein contents of BAT-Exos were analyzed by mass spectrometry. Results: The results showed that BAT-Exos reduced the body weight, lowered blood glucose and alleviated lipid accumulation in HFD mice independently of food intake. Echocardiography revealed that the abnormal cardiac functions of HFD mice were significantly restored after treatment with BAT-Exos. Cell metabolism analysis showed that treatment with BAT-Exos significantly promoted oxygen consumption in recipient cells. Protein profiling of exosomes demonstrated that BAT-Exos were rich in mitochondria components and involved in catalytic processes. Conclusions: Collectively, our study showed that BAT-Exos significantly mitigated the metabolic syndrome in HFD mice. Detailed elucidation of the reactive molecules and mechanism of action would provide new insights in combating obesity and related disorders.

A new strategy for accurate targeted diagnosis and treatment of cutaneous malignant melanoma: dual-mode phase-change lipid nanodroplets as ultrasound contrast agents.

BACKGROUND: Currently, effective detection and treatment of cutaneous malignant melanoma (CMM) still face severe challenges. Ultrasound molecular imaging as a noninvasive and easy-to-operate method is expected to bring improvements for tumor detection. PURPOSE: The aim of this research is to prepare novel phase-change ultrasound contrast agents, Nds-IR780, which can perform not only dual-mode molecule-targeted imaging but also targeted photothermal therapy for CMM. METHODS: A double emulsion process was used to prepare the Nds-IR780. Then, the entrapment rate and drug loading of IR-780 iodide in Nds-IR780 were detected by high-performance liquid chromatography. The biocompatibility of Nds-IR780 was evaluated by a CCK-8 assay and the characteristics and stability of that were verified through the particle size analyzer, laser scanning confocal microscopy (LSCM) and transmission electron microscopy (TEM). The abilities of dual-mode molecule-targeted imaging and targeted photothermal therapy for Nds-IR780 were confirmed via the in vitro and in vivo experiments. RESULTS: Nds-IR780 had good size distribution, polydispersity index, stability and biosafety. The in vitro and in vivo experiments confirmed that Nds-IR780 were capable of targeting CMM cells with high affinity (22.4±3.2%) and facilitating dual-mode imaging to detect the primary lesion and sentinel lymph nodes (SLNs) of CMM. Furthermore, the photothermal ablation of CMM mediated by Nds-IR780 was very effective in vivo. CONCLUSION: The newly prepared Nds-IR780 were observed to be effective targeted theranostic probe for the precise detection and targeted treatment of CMM.

Intraoperative contrast-enhanced ultrasound for cerebral glioma resection and the relationship between microvascular perfusion and microvessel density.

We analyzed the relationship between quantitative CEUS parameters and microvessel density (MVD) of different pathologic grades of cerebral gliomas. ICEUS was performed in 49 patients with cerebral gliomas. The enhancement characteristics of cerebral gliomas were observed before and after tumor resection. The number of microvessels was counted by immunostaining with anti-CD34. Differences in these quantitative parameters in cerebral gliomas were compared and subjected to a correlation analysis with MVD. The assessment of iCEUS parameters and tumor MVD showed that cerebral gliomas of different pathological grades had different characteristics. The time-to-peak (Tmax) was significantly shorter, the peak intensity (PI) and MVD were significantly higher in high-grade cerebral gliomas than in low-grade cerebral gliomas (p < 0.05). According to the immunostaining, PI was positively (r = 0.637) correlated with MVD and Tmax was negatively (r = -0.845) correlated with MVD. ICEUS could provid dynamic and continuous real-time imaging and quantitative data analysis of different pathological grades of cerebral gliomas, the quantitiative CEUS parameters were closely related to the MVD, and be helpful in understanding the cerebral gliomas grade and refining surgical strategy.

Stiffening of aorta is more preferentially associated with rheumatoid arthritis than peripheral arteries.

The objective of this study is to investigate the relative impact of rheumatoid arthritis (RA) and other factors on arterial stiffness of different regions assessed by regional pulse wave velocity (PWV). Seventy-two patients with RA and 55 strictly matched healthy controls were included. Doppler ultrasound was used to measure the PWV of heart-carotid (hcPWV), heart-femoral (hfPWV), brachial-radial (brPWV), femoral-ankle (faPWV) and carotid-femoral segments (cfPWV) in all subjects. The reproducibility of regional PWV measurement was evaluated in 30 random RA patients. In RA patients, the hfPWV and cfPWV were significantly higher than that in controls (P = 0.0006, P = 0.0001, respectively), and the hcPWV, brPWV and faPWV only showed an increase trend without significance. The mean increase magnitude of hfPWV (17.5%) and cfPWV (18.5%) were greater than brPWV (7.2%) and faPWV (1.7%) in RA patients. The association between RA and both hfPWV, cfPWV remained significant after adjustments for other confounders (P < 0.001). However, the association between RA and brPWV (P = 0.199), faPWV (P = 0.599) was not significant. In addition, age and systolic blood pressure were also significant independent factors associated with hfPWV and cfPWV. The reproducibility analysis showed that hfPWV and cfPWV measurements had lower coefficient of variation than others. The stiffness of different arterial regions is not equally affected by RA. The stiffening of aorta is more preferentially associated with RA than that of the peripheral arteries in extremities. The discrepant stiffening between aorta and peripheral arteries may provide a new insight into the pathogenesis of cardiovascular and microvascular dysfunction frequently occurred in RA.