Epidemiological Trends in Cardiovascular Disease Mortality Attributable to Modifiable Risk Factors and Its Association with Sociodemographic Transitions across BRICS-Plus Countries.

BRICS-Plus countries (Brazil, Russia, India, China, South Africa, and 30 other countries) is a group of 35 countries with emerging economies making up more than half of the world's population. We explored epidemiological trends of cardiovascular disease (CVD) mortality attributable to modifiable risk factors and its association with period and birth cohort effects and sociodemographic index (SDI) across BRICS-Plus countries by using joinpoint regression and age-period-cohort modeling from 1990 to 2019. Between 1990 and 2019, the all-ages CVD deaths increased by 85.2% (6.1 million to 11.3 million) across BRICS-Plus countries. The CVD age-standardized mortality rate attributable to dietary risks and smoking significantly decreased across BRICS-Plus countries, with some exceptions. However, four-fifths of BRICS-Plus countries observed a remarkable increasing trend of high body mass-index (BMI)-related CVD deaths, in particular, among younger adults (25-49 years). Early birth cohorts and individuals aged greater than 50 years showed a higher risk of CVD mortality. Both the China-ASEAN FTA and Mercosur regions stand out for their successful sociodemographic transition, with a significant reduction in CVD mortality over the study period. Singapore and Brazil achieved great progress in CVD mortality reduction and the other BRICS-Plus countries should follow their lead in adopting public health policies and initiatives into practice.

An "AND" Molecular Logic Gate as a Super-Enhancers for De Novo Designing Activatable Probe and Its Application in Atherosclerosis Imaging.

Developing activatable fluorescent probes with superlative fluorescence enhancement factor (F/F0 ) to improve the signal-to-noise (S/N) ratio is still an urgent issue. "AND" molecular logic gates are emerging as a useful tool for enhanced probes selectivity and accuracy. Here, an "AND" logic gate is developed as super-enhancers for designing activatable probes with huge F/F0 and S/N ratio. It utilizes lipid-droplets (LDs) as controllable background input and sets the target analyte as variable input. The fluorescence is tremendously quenching due to double locking, thus an extreme F/F0 ratio of target analyte is obtained. Importantly, this probe can transfer to LDs after a response occurs. The target analyte can be directly visualized through the spatial location without a control group. Accordingly, a peroxynitrite (ONOO- ) activatable probe (CNP2-B) is de novo designed. The F/F0 of CNP2-B achieves 2600 after reacting with ONOO- . Furthermore, CNP2-B can transfer from mitochondria to lipid droplets after being activated. The higher selectivity and S/N ratio of CNP2-B are obtained than commercial probe 3'-(p-hydroxyphenyl) fluorescein (HPFin vitro and in vivo. Therefore, the atherosclerotic plaques at mouse models are delineated clearly after administration with in situ CNP2-B probe gel. Such input controllable "AND" logic gate is envisioned to execute more imaging tasks.

Peroxynitrite/Lipid Droplet Sequence-Activated Dual-Lock Fluorescent Probes Enable Precise Intraoperative Imaging of Atherosclerotic Plaques.

The formation of atherosclerotic plaques is the root cause of various cardiovascular diseases (CVDs). Effective CVD interventions thus call for precise identification of the plaques to aid clinical assessment and treatment of such diseases. In this study, we introduced a dual-analyte sequentially activated logic fluorescence reporting system CNN2-B to precisely identify the atherosclerotic plaques in vivo. This probe was achieved by creating a dual-locked fluorescent sensor that permits highly specific and sensitive detection of peroxynitrite and lipid droplets─the two hallmarks of atherosclerosis (AS). The recognition group of the probe removed after reacting with ONOO- and intramolecular charge rearrangement occurred to generate a coumarin derivative structure. This structure had a strong solvent effect; it could recognize lipid droplets (LDs) in cells, thus exhibiting fluorescence without secondary molecular adjustment. The fluorescence was tremendously quenched by double locking; thus, an extreme fluorescence enhancement factor (F/F0) ratio of 365 for CNN2-B was obtained. Importantly, CNN2-B could move from the mitochondria to lipid droplets after being activated. CNN2-B exhibited higher selectivity and signal-to-noise (S/N) ratio than commercial probe hydroxyphenyl fluorescein (HPF). Therefore, atherosclerotic plaques in mouse models were delineated clearly by fluorescence imaging after in situ administration of CNN2-B.

Enzyme Catalysis Biomotor Engineering of Neutrophils for Nanodrug Delivery and Cell-Based Thrombolytic Therapy.

Utilizing neutrophils (NEs) to target and deliver nanodrugs to inflammation sites has received considerable attention. NEs are involved in the formation and development of thrombosis by transforming into neutrophil extracellular traps (NETs); this indicates that NEs may be a natural thrombolytic drug delivery carrier. However, NEs lack an effective power system to overcome blood flow resistance and enhance targeting efficiency. Herein, we report the application of a urease catalysis micromotor powered NEs nanodrug delivery system to promote thrombolysis and suppress rethrombosis. The urease micromotor powered Janus NEs (UM-NEs) were prepared by immobilizing the enzyme asymmetrically onto the surface of natural NEs and then loading urokinase (UK) coupled silver (Ag) nanoparticles (Ag-UK) to obtain the UM-NEs (Ag-UK) system. Urease catalytic endogenous urea is used to generate thrust by producing ammonia and carbon dioxide, which propels NEs actively targeting the thrombus. The UM-NEs (Ag-UK) can be activated by enriched inflammatory cytokines to release NETs at the thrombosis site, resulting in a concomitant release of Ag-UK. Ag-UK induces thrombolysis to restore vascular recanalization. This urease micromotor-driven NEs drug delivery system can significantly reduce the hemorrhagic side effects, promote thrombolysis, and inhibit rethrombosis with high bioavailability and biosafety, which can be used for the treatment of thrombotic diseases.

Lipid-Activatable Fluorescent Probe for Intraoperative Imaging of Atherosclerotic Plaque Using In Situ Patch.

The surgical removal of lesions is among the most common and effective treatments for atherosclerosis. It is often the only curative treatment option, and the ability to visualize the full extent of atherosclerotic plaque during the operation has major implications for the therapeutic outcome. Fluorescence imaging is a promising approach for the inspection of atherosclerotic plaques during surgery. However, there is no systematic strategy for intraoperative fluorescent imaging in atherosclerosis. In this study, the in situ attachment of a lipid-activatable fluorescent probe (CN-N2)-soaked patch to the outer arterial surface is reported for rapid and precise localization of the atherosclerotic plaque in ApoE-deficient mouse during surgery. Stable imaging of the plaque is conducted within 5 min via rapid recognition of abnormally accumulated lipid droplets (LDs) in foam cells. Furthermore, the plaque/normal ratio (P/N) is significantly enhanced to facilitate surgical delineation of carotid atherosclerotic plaques. Visible fluorescence bioimaging using lipid-activatable probes can accurately delineate plaque sizes down to diameters of <0.5 mm, and the images can be swiftly captured within the stable plaque imaging time window. These findings on intraoperative fluorescent imaging of plaques via the in situ attachment of the CN-N2 patch hold promise for effective clinical applications.

Lipid Droplet-Specific Probe for Rapidly Locating Atherosclerotic Plaques and Intraoperative Imaging via In Situ Spraying.

The ability to visualize the full extent of atherosclerotic plaques during surgery has major implications for therapeutic outcomes. Fluorescence imaging is a promising approach for atherosclerotic plaque inspection during surgery. However, a specific strategy for the intraoperative fluorescence imaging of atherosclerosis has not been established. This study presents an in situ spraying aerosol of a lipid droplet-specific probe to rapidly and precisely locate atherosclerotic plaques during surgery. Stable imaging of the plaque was achieved within 5 min by nebulizing the aqueous solution of the lipid droplet-specific probe (CN-PD) into 3 µm droplets and rapidly permeating it in situ. The visible fluorescence bioimaging of CN-PD can accurately delineate the plaque margins and size even with a diameter ≤0.5 mm, which are capable of being swiftly captured during the stable plaque imaging window (>2 h). This strategy combines the consideration of a specific probe design and an efficient in situ delivery, which results in weak interference from the background signals. Therefore, the plaque-to-normal tissue ratio (P/N) is sufficient to facilitate the surgical delineation of carotid atherosclerotic plaques. The originality of the intraoperative fluorescence imaging of the plaques via in situ delivery of the lipid droplet-specific probe holds promise for effective clinical application.

Controlled "off-on" fluorescent probe for the specific detection of hyperhomocysteinemia.

Hyperhomocysteinemia is an established risk factor for atherosclerosis and vascular disease. Therefore, designing a hyperhomocysteinemia specific probe is of great significance for the early warning of cardiovascular diseases. However, developing probes that can efficiently and specifically recognize homocysteine (Hcy) remains a tremendous challenge. Therefore, we designed an Hcy-specific fluorescent probe (HSFP) with excellent selectivity and anti-interference capability. Interestingly, this probe can automatically "off-on" in water solution, but the fluorescence of HSFP remains "off" when Hcy is present in the solution. The spectroscopic data demonstrated that the fluorescence of HSFP attenuated 13.8 folds toward Hcy in water without interference from other biothiols and amino acids. Furthermore, HSFP can sensitively reflect the change of Hcy content in cells. Therefore, HSFP was further applied to detect hyperhomocysteinemia in vivo with high efficiency. In summary, we have developed an Hcy-specific fluorescent probe to efficiently detect Hcy in vivo and in vitro, which may contribute to basic or clinical research.

A Bioresponsive Near-Infrared Fluorescent Probe for Facile and Persistent Live-Cell Tracking.

Molecular imaging significantly transforms the field of biomedical science and facilitates the visualization, characterization, and quantification of biologic processes. However, it is still challenging to monitor cell localization in vivo, which is essential to the study of tumor metastasis and in the development of cell-based therapies. While most conventional small-molecule fluorescent probes cannot afford durable cell labeling, transfection of cells with fluorescent proteins is limited by their fixed fluorescence, poor tissue penetration, and interference of autofluorescence background. Here, a bioresponsive near-infrared fluorescent probe is reported as facile and reliable tool for real-time cell tracking in vivo. The design of this probe relies on a new phenomenon observed upon fluorobenzene-conjugated fluorescent dyes, which can form complexes with cytosolic glutathione and actively translocates to lysosomes, exhibiting enhanced and stable cell labeling. Fluorobenzene-coupled hemicyanine, a near-infrared fluorophore manifests to efficiently staining tumor cells without affecting their invasive property and enables persistent monitoring of cell migration in metastatic tumor murine models at high resolution for one week. The method of fluorobenzene functionalization also provides a simple and universal "add-on" strategy to render ordinary fluorescent probes suitable for long-term live-cell tracking, for which currently there is a deficit of suitable molecular tools.

A new lysosome-targetable fluorescent probe for detection of endogenous hydrogen polysulfides in living cells and inflamed mouse model.

Hydrogen polysulfides (H2Sn, n > 1) belong to sulfane sulfur in the reactive sulfur species (RSS) family and play significant roles in maintaining biological homeostasis in organisms. The detection of H2Sn in living systems is essential, but further understanding of its "intact" function in living cells has been limited, owing to the lack of appropriate analytical tools. In this work, a new fluorescent probe PP-PS was designed for the detection of endogenous H2Sn. The probe has a large Stokes shift (178 nm), low detection limit (1 nM), and short response time (1 minute). Besides, PP-PS was successfully applied in the imaging of endogenous H2Sn in lysosomes of living cancer cells, xenograft mouse tumor tissues, and LPS-induced mouse inflammation tissues. These results revealed that the probe PP-PS could act as a new fluorescence imaging tool to study the function of intracellular hydropolysulfides.

A family of push-pull bio-probes for tracking lipid droplets in living cells with the detection of heterogeneity and polarity.

Lipid droplets (LDs) are multi-functional organelles with the storage of lipid and participating in a variety of physiological processes, including membrane transport and signal transduction. The dysfunction of LDs has been reported to be associated with multiple diseases such as obesity, diabetes, atherosclerosis, and cancer in research. Herein, we designed and synthesized a family of push-pull bio-probes (LDP1-LDP4) based on thiophene or 3,4-ethylenedioxythiophene, which is also called EDOT. LDP1-LDP4 showed positive solvatochromic effect from toluene to ethanol and the maximum fluorescence wavelength redshifted to 165 nm. It was found that the four probes showed significant increase in fluorescence intensity from PBS to oil. LDP1-LDP4 displayed excellent biocompatibility and good optical properties and had substantially facilitated to track LDs with detecting heterogeneity. LDP4 was also used to expose the difference in the polarity of LDs and cytoplasm.

Application of Nitroimidazole-Carbobane-Modified Phenylalanine Derivatives as Dual-Target Boron Carriers in Boron Neutron Capture Therapy.

Boron neutron capture therapy (BNCT) has received extensive attention as noninvasive cell-level oncotherapy for treating solid cancer tumors. However, boron-containing drugs such as l-boronophenylalanine (BPA) and sodium borocaptate have low boron content and/or poor tumor-targeting ability, limiting their application. In this study, we designed and synthesized a series of nontoxic, dual-target boron carriers (B139, B142, and B151) with the ability to accumulate specifically in tumor cells. We found that the B139 uptake into hypoxic tumor regions was high, with a 70-fold boron content compared to BPA. In addition, in vivo observation showed that B139 can be trapped in tumor cells for a prolonged period and maintains an effective therapeutic concentration, with a peak boron concentration of 50.7 µg/g and a high tumor: blood boron ratio of >3, achieving ideal BNCT conditions. Cytotoxicity evaluation in mice further proved that B139 is safe and reliable. Therefore, B139 has great potential for BNCT application as a dual-target, safe, and efficient boron carrier.

Correction: A tumor-targeting probe based on a mitophagy process for live imaging.

Correction for 'A tumor-targeting probe based on a mitophagy process for live imaging' by Lijuan Gui et al., Chem. Commun., 2018, 54, 9675-9678.

Near-infrared off-on fluorescence probe activated by NTR for in vivo hypoxia imaging.

Selective and efficient detection and imaging of nitroreductase (NTR) overexpressed in hypoxic tissues is of great importance for better understanding its biological functions. The effective optical probes equipped for NTR detection in vivo is still lacking, so we developed an innovative near infrared (NIR) fluorescent on-off probe (Cy-NO2) composed of fluorescence reporting unit (an aminocyaine dye) and recognition unit (p-nitrobenzylcarbamate group). The response and mechanism of the NTR activated reduction of Cy-NO2 were evaluated in vitro through kinetic optical studies, mass spectra analysis and docking calculations. The results demonstrated that Cy-NO2 could rapidly recognize NTR with high selectivity and sensitivity. The efficient NTR detection ability of Cy7-NO2 was further validated by fluorescence imaging of hypoxic cells (A549, PC-12 and HUVEC cell). Furthermore, the in vivo hypoxia imaging by Cy-NO2 were evaluated on tumor-bearing mouse, cerebral ischemia (CIS) and deep vein thrombosis models. The results indicated a rapid and distinct enhancement of its NIR fluorescence highly appropriate for in vivomonitoring of NTR in all of the three animal models, which revealed aspiring clinical value of this NIR fluorescent hypoxia probe.

GSH-Activated Light-Up Near-Infrared Fluorescent Probe with High Affinity to αvß3 Integrin for Precise Early Tumor Identification.

The development of tumor-associated, stimuli-driven, turn-on near-infrared (NIR) fluorophores requires urgent attention because of their potential in selective and precise tumor diagnosis. Herein, we describe a NIR fluorescent probe (CyA-cRGD) comprised of a fluorescence reporting unit (a cyanine dye) linked with a GSH-responsive unit (nitroazo aryl ether group) and a tumor-targeting unit (cRGD). The NIR fluorescence of CyA-cRGD with sensitive and selective response to GSH can act as a direct off-on signal reporter for GSH monitoring. Notably, CyA-cRGD possesses improved biocompatibility compared with CyA, which is highly desirable for in vivo fluorescence tracking of cancer. Confocal fluorescence imaging confirmed the tumor-targeting capability and GSH detection ability of CyA-cRGD in tumor cells, normal cells, and coincubated tumor /normal cells and in the three-dimensional multicellular tumor spheroid. Furthermore, it was validated that CyA-cRGD could detect tumor precisely in GSH and integrin αvß 3 high-expressed tumor-bearing mouse models. Importantly, it was confirmed that CyA-cRGD possessed high efficiency for early-stage tumor imaging in mouse models with tumor cells implanted within 72 h. This method provided significant advances toward more in-depth understanding and exploration of tumor imaging, which may potentially be applied for clinical early tumor diagnosis.

A tumor-targeting probe based on a mitophagy process for live imaging.

A glucosamine modified near-infrared cyanine dye CyT sensitive to pH was synthesized. Due to the different pH values of mitochondria and autolysosomes, the probe can simultaneously investigate mitochondria and autolysosomes in living cells. Moreover, due to the introduction of glucosamine groups, this fluorescent probe can be applied for tumor targeted imaging.

Inhibition of microRNA-299-5p sensitizes glioblastoma cells to temozolomide via the MAPK/ERK signaling pathway.

Glioblastomas (GBMs) are a lethal class of brain cancer, with a median survival <15 months in spite of therapeutic advances. The poor prognosis of GBM is largely attributed to acquired chemotherapy resistance, and new strategies are urgently needed to target resistant glioma cells. Here we report a role for miR-299-5p in GBM. The level of miR-299-5p expression was detected in glioma specimens and cell lines by qRT-PCR. Luciferase reporter assays and Western blots were performed to verify GOLPH3 as a direct target of miR-299-5p. In vitro cell proliferation, invasion, cell cycle distribution, and apoptosis were assessed to determine whether or not miR-299-5p knockdown sensitized GBM cells to temozolomide (TMZ). We demonstrated that miR-299-5p levels were up-regulated in the GBM groups compared with the normal control group. The highest expression of miR-129-5p occurred in the highest GBM stage. miR-299-5p knockdown significantly inhibited the MAPK/extracellular signal-regulated kinase (ERK) signaling pathway. We also showed that miR-299-5p knockdown enhanced sensitivity of GBM cells to TMZ both in vitro and in vivo by inhibiting cell proliferation and invasion and promoting apoptosis. In addition, we demonstrated that GOLPH3 is a novel functional target of miR-299-5p GOLPH3 regulates the MAPK/ERK axis under miR-299-5p regulation. In conclusion, we identified a link between miR-299-5p expression and the GOLPH3/MAPK/ERK axis, thus illustrating a novel role for miR-299-5p in GBM.

Methionine-Decorated Near Infrared Fluorescent Probe for Prolonged Tumor Imaging.

Methionine (Met) is one of the essential amino acids of which the transport system L is overexpressed in various tumor cells. In this study, a near-infrared fluorescent dye (IR-780) and methionine were conjugated through a piperazin-polyamines linker to form Cy-Met. The successful synthesis of Cy-Met was validated by optical characterization, NMR, and MS spectra. The absorption peak of Cy-Met was at 680 nm, and the fluorescence peak was at 790 nm. The cytotoxicity assay and cell imaging studies indicated that Cy-Met had good biocompatibility and specific affinity to tumor cells. The dynamic distribution and clearance investigations showed that Cy-Met was eliminated by the liver-intestine pathway. Notably, Cy-Met displayed tumor-targeting ability in U87, H22, and EAC tumor-bearing mice with an evident long circulation time. The results implied that Cy-Met could act as a promising fluorescence probe specialized for long-term tumor monitoring.

Biocompatible tumor-targeting nanocomposites based on CuS for tumor imaging and photothermal therapy.

Active targeting of tumor receptors is a significant approach for cancer diagnosis. Additionally, development of photothermal agents for photothermal therapy (PTT) has attracted great interest in the field of nanomedicine. In the present study, copper sulfide (CuS) nanoparticles capped with bovine serum albumin (BSA), named CuS@BSA, was synthesized by a convenient method. Then, the near-infrared (NIR) fluorescence probe MBA and the tumor-targeting ligand cyclic RGD were further conjugated on the surface of CuS@BSA, and the obtained nanocomposite was named CuS@BSA-MBA-cRGD. The morphology, optical properties, biotoxicity, tumor-targeting capability and in vitro and in vivo tumor inhibition effect were all characterized comprehensively. This nanocomplex demonstrated enhanced photothermal effects and positive tumor targeting. Thus, the nanocomposite CuS@BSA-MBA-cRGD can used as a promising tumor-targeting PTT agent for simultaneous cancer imaging and photothermal treatment.

The saponin D39 blocks dissociation of non-muscular myosin heavy chain IIA from TNF receptor 2, suppressing tissue factor expression and venous thrombosis.

BACKGROUND AND PURPOSE: Non-muscular myosin heavy chain IIA (NMMHC IIA) plays a key role in tissue factor expression and venous thrombosis. Natural products might inhibit thrombosis through effects on NMMHC IIA. Here, we have shown that a natural saponin, D39, from Liriope muscari exerted anti-thrombotic activity in vivo, by targeting NMMHC IIA. EXPERIMENTAL APPROACH: Expression and activity of tissue factor in endothelial cells were analysed in vitro by Western blot and simplified chromogenic assays. Interactions between D39 and NMMHC IIA were assessed by serial affinity chromatography and molecular docking analysis. D39-dependent interactions between NMMHC IIA and TNF receptor 2 (TNFR2) were measured by immunofluorescence, co-immunoprecipitation and proximity ligation assays. Anti-thrombotic activity of D39 in vivo was evaluated with a model of inferior vena cava ligation injury in mice. KEY RESULTS: D39 inhibited tissue factor expression and procoagulant activities in HUVECs and decreased thrombus weight in inferior vena cava-ligated mice dose-dependently. Serial affinity chromatography and molecular docking analysis suggested that D39 bound to NMMHC IIA. In HEK293T cells, D39 inhibited tissue factor expression evoked by NMMHC IIA overexpression. This effect was blocked by NMMHC IIA knockdown in HUVECs. D39 inhibited dissociation of NMMHC IIA from TNFR2, which subsequently modulated the Akt/GSK3ß-NF-κB signalling pathways. CONCLUSIONS AND IMPLICATIONS: D39 inhibited tissue factor expression and thrombus formation by modulating the Akt/GSK3ß and NF-κB signalling pathways through NMMHC IIA. We identified a new natural product that targeted NMMHC IIA, as a potential treatment for thrombotic disorders and other vasculopathies.