Self-Assembled Metal-Coordination Nanohelices as Efficient and Robust Chiral Supramolecular Catalysts for Enantioselective Reactions.

The development of chiral nanostructures-based supramolecular catalysts with satisfied enantioselectivity remains a significantly more challenging task. Herein, the synthesis and self-assembly of various amino acid amphiphiles as chiral supramolecular catalysts after metal ion coordination is reported and systematically investigate their enantioselectivity in asymmetric Diels-Alder reactions. In particular, the self-assembly of l/d-phenylglycine-based amphiphiles (l/d-PhgC16) and Cu(II) into chiral supramolecular catalysts in the methanol/water solution mixture is described, which features the interesting M/P nanohelices (diameter ≈8 nm) and mostly well-aligned M/P nanoribbons (NRs). The M/P supramolecular catalysts show both high but inverse enantioselectivity (>90% ee) in Diels-Alder reactions, while their monomeric counterparts display nearly racemic products. Analysis of the catalytic results suggests the outstanding enantioselectivities are closely related to the specific stereochemical microenvironment provided by the arrangement of the amphiphiles in the supramolecular assembly. Based on the experimental evidence of chirality transfer from supramolecular nanohelices to coordinated Cu(II) and substrate aza-chalcone and the molecular dynamics simulations, the enantioselective catalytic mechanisms are proposed. Moreover, the relationships between molecular structures of amino acid amphiphiles (the hydrophilic head group and hydrophobic alkyl chain length) in supramolecular catalysts and enantioselectivity in Diels-Alder reactions are elaborated.

FTO-mediated ZNF687 accelerates tumor growth, metastasis, and angiogenesis in colorectal cancer through the Wnt/ß-catenin pathway.

Colorectal cancer (CRC) is a common and lethal cancer. ZNF687 has been disclosed to take part in diversified cancers' progression by serving as a facilitator. However, the detailed regulatory functions of ZNF687 in the CRC have not been investigated. This work is planned to probe the impacts of ZNF687 on CRC progression. The IHC, RT-qPCR, and western blot assays were used to examine mRNA and protein gene expressions. The cell proliferation measurement was accompanied by a CCK-8 assay. The Transwell assay was performed to evaluate cell invasion and migration. The angiogenesis ability was evaluated by a tube formation experiment. The m6A level was evaluated through MeRIP and m6A dot blot assays. The binding ability between ZNF687 and FTO (fat mass and obesity associated protein) was tested through an RIP assay. The ß-catenin nuclear translocation was assessed through an immunofluorescence assay. The tumor growth was evaluated through an in vivo assay. ZNF687 exhibited higher expression in CRC cells and resulted in a poor prognosis. Additionally, ZNF687 inhibition suppressed CRC cell proliferation, invasion, migration, and angiogenesis. Furthermore, the suppression of ZNF687 retarded the Wnt pathway. Through rescue assays, the reduced cell migration, proliferation, invasion, and angiogenesis mediated by ZNF687 knockdown could be reversed after BML-284 (the activator of the Wnt pathway) treatment. Finally, it was explained that ZNF687 knockdown inhibited in vivo tumor growth. This study manifested that FTO-mediated ZNF687 aggravated tumor growth, metastasis, and angiogenesis of CRC through Wnt/ß-catenin pathway. This finding may provide a hopeful molecular target for CRC treatment.

Ras Inhibitor Lonafarnib Rescues Structural and Functional Impairments of Synapses of Aß1-42 Mice via α7nAChR-Dependent BDNF Upregulation.

Alzheimer's disease (AD) is characterized pathologically by the structural and functional impairments of synapses in the hippocampus, inducing the learning and memory deficiencies. Ras GTPase is closely related to the synaptic function and memory. This study was to investigate the effects of farnesyl transferase inhibitor lonafarnib on the synaptic structure and function in AD male mice and explore the potential mechanism. Our results showed 50 mg/kg lonafarnib (intraperitoneal) rescued the impaired spatial memory and improved the damaged synaptic transmission and plasticity of Aß1-42 mice. In addition, lonafarnib ameliorated the morphology of synaptic dendrites and spines in Aß1-42 mice. Furthermore, lonafarnib enhanced α7nAChR cell surface expression and phosphorylation of downstream Akt and CaMKII in Aß1-42 mice, which were inhibited by α7nAChR antagonist methyl lycaconitine (MLA), and increased the phosphorylation of CREB in a CaMKII- but not ERK-dependent way. Lonafarnib enhanced hippocampal brain-derived neurotrophic factor (BDNF) concentration in Aß1-42 mice, which was sensitive to MLA and KN93 (an inhibitor of CaMKII), but not related to ERK and Akt pathways. H-Ras, but not Rhes, was related to the lonafarnib induced improvement of α7nAChR cell surface expression and BDNF content. Interestingly, lonafarnib induced improvement of synaptic transmission, plasticity and spatial cognition in Aß1-42 mice was abolished by BDNF deprivation with TrkB/Fc chimera protein. Our results indicate that lonafarnib can rescue the structural and functional impairments of synapses in the Aß1-42 mice, which may be related to the improvement of BDNF content through the H-Ras-α7nAChR-dependent CaMKII-CREB pathway, leading to the improvement of spatial cognition.SIGNIFICANCE STATEMENT Alzheimer's disease (AD) is characterized pathologically by the structural and functional impairments of synapses in the hippocampus, inducing the learning and memory deficiencies. However, no effective drugs have not been developed for the treatment of AD synaptic. This study for the first time reported the beneficial effects of Ras inhibitor lonafarnib on the synaptic structure and function in AD mice, providing an alternative way for the treatment of "synaptic disease" in AD patients.

Supramolecular Polyaniline-Metal Ion as Chiral Nanozymes for Enantioselective Catalysis.

Understanding origin of asymmetric information encoded on chiral nanozymes is important in mediating enantioselective catalysis. Herein, the supramolecular chiral nanozymes constructed from P/M-polyaniline (P/M-PANI) nanotwists and metal ions (M2+ , M = Cu, Ni, Co, and Zn) are designed through thioglycolic acid (TA) without chiral molecules to show the regulated catalytic efficiency and enantioselectivity. With combination of chiral environment from supramolecular scaffolds and catalytic center from metal ions, the P-PANI-TA-M2+ as nanozymes show preference to 3,4-dihydroxy-S-phenylalanine (S-DOPA) oxidation while the M-PANI-TA-M2+ show better selectivity to R-DOPA oxidation. Among them, though the Cu2+ doped supramolecular nanotwists show the highest catalytic efficiency, the Co2+ doped ones with moderate catalytic efficiency can exhibit the best enantioselectivity with select factor as high as 2.07. The molecular dynamic (MD) simulation clarifies the mechanism of enantioselective catalysis caused by the differential kinetics with S/R-DOPA enantiomers adsorbed on chiral PANI surface and free in solution. This work systematically studies the synergistic effect between the chiral supramolecular nanostructures assembled by achiral species and metal ions as peroxidase-like catalytic centers to regulate the enantioselectivity, providing deep understanding of the origin of asymmetric catalysis and serving as strong foundation to guide the design of nanozymes with high enantioselectivity.

Phosphatidylinositol 4,5-Bisphosphate Sensing Lipid Raft via Inter-Leaflet Coupling Regulated by Acyl Chain Length of Sphingomyelin.

Phosphatidylinositol 4,5-bisphosphate (PIP2) is an important molecule located at the inner leaflet of cell membrane, where it serves as anchoring sites for a cohort of membrane-associated molecules and as a broad-reaching signaling intermediate. The lipid raft is thought as the major platform recruiting proteins for signal transduction and also known to mediate PIP2 accumulation across the membrane. While the significance of this cross-membrane coupling is increasingly appreciated, it remains unclear whether and how PIP2 senses the dynamic change of the ordered lipid domains over the packed hydrophobic core of the bilayer. Herein, by means of molecular dynamic simulation, we reveal that inner PIP2 molecules can sense the outer lipid domain via inter-leaflet coupling, and the coupling manner is dictated by the acyl chain length of sphingomyelin (SM) partitioned to the lipid domain. Shorter SM promotes membrane domain registration, whereby PIP2 accumulates beneath the domain across the membrane. In contrast, the anti-registration is thermodynamically preferred if the lipid domain has longer SM due to the hydrophobic mismatch between the corresponding acyl chains in SM and PIP2. In this case, PIP2 is expelled by the domain with a higher diffusivity. These results provide molecular insights into the regulatory mechanism of correlation between the outer lipid domain and inner PIP2, both of which are critical components for cell signal transduction.

A deep learning-enabled smartphone platform for rapid and sensitive colorimetric detection of dimethoate pesticide.

A novel deep learning-enabled smartphone platform is developed to assist a colorimetric aptamer biosensor for fast and highly sensitive detection of dimethoate. The colorimetric determination of dimethoate is based on the specific binding of dimethoate and aptamer, which leads to the aggregation of AuNPs in high-concentration NaCl solution, resulting in an obvious color change from red to blue. This color change provides sufficient data for self-learning enabled by a convolutional neural network (CNN) model, which is established to predict dimethoate concentration based on images acquired from a smartphone. To enhance user-friendliness for non-experts, the CNN model is then embedded into a smartphone app, enabling offline detection of dimethoate pesticide in real environments within just 15 min using a pre-configured colorimetric probe. The developed platform exhibits superior performance, achieving a regression coefficient of 0.9992 in the concentration range of 0-10 µM. Moreover, the app's performance is found to be consistent with the ELISA kit. These remarkable findings demonstrate the potential of combining colorimetric biosensors with smartphone-based deep learning methods for the development of portable and affordable tools for pesticide detection.

De novo upper tract urothelial carcinoma after renal transplantation: a single-center experience in China.

BACKGROUND: Long-term prognosis and risk factors of de novo upper tract urothelial carcinoma after renal transplantation were rarely studied. Thus, the aim of this study was to investigate the clinical features, risk factors, and long-term prognosis of de novo upper tract urothelial carcinoma after renal transplantation, especially the impact of aristolochic acid on tumor, using a large sample. METHODS: 106 patients were enrolled in retrospective study. The endpoints included overall survival, cancer-specific survival, bladder or contralateral upper tract recurrence-free survival. Patients were grouped according to aristolochic acid exposure. Survival analysis was performed using Kaplan-Meier curve. Log-rank test was used to compare the difference. Multivariable cox regression was conducted to evaluate the prognostic significance. RESULTS: Median time from transplantation to development of upper tract urothelial carcinoma was 91.5 months. Cancer-specific survival rate at 1, 5, 10 years was 89.2%, 73.2%, 61.6%. Tumor staging (≥ T2), lymph node status (N +) were independent risk factors for cancer-specific death. Contralateral upper tract recurrence-free survival rate at 1, 3, 5 years was 80.4%, 68.5%, 50.9%. Aristolochic acid exposure was independent risk factor for contralateral upper tract recurrence. The patients exposed to aristolochic acid had more multifocal tumors and higher incidence of contralateral upper tract recurrence. CONCLUSION: Both higher tumor staging and positive lymph node status were associated with a worse cancer-specific survival in patients with post-transplant de novo upper tract urothelial carcinoma, which highlighted the importance of early diagnosis. Aristolochic acid was associated with multifocality of tumors and higher incidence of contralateral upper tract recurrence. Thus, prophylactic contralateral resection was suggested for post-transplant upper tract urothelial carcinoma, especially for patients with aristolochic acid exposure.

Protein-enriched extracts from housefly (Musca domestica) maggots alleviates atherosclerosis in apolipoprotein E-deficient mice by promoting bile acid production and consequent cholesterol consumption.

Currently, atherosclerosis control is important to prevent future heart attacks or strokes. Protein-enriched extract (PE) from housefly maggots (Musca domestica) can inhibit the development of atherosclerosis partially through its antioxidant effects. Whether PE exerts other anti-atherosclerosis functions remains unclear. Here, PE was found to simultaneously promote cholesterol metabolism effects in apolipoprotein E knockout (ApoE-/- ) mice. Bile acid synthesis plays a key role in regulating cholesterol homeostasis in atherosclerosis. Whether PE alleviates atherosclerosis by promoting bile acid production and consequent cholesterol consumption was further explored. First, 8-week-old male ApoE-/- mice were recruited and fed on a cholesterol-enriched diet. After 8 weeks, these mice were divided into three groups and received gavage administration of PE, simvastatin, and saline for another 8 weeks. Atherosclerosis severity was then assessed. Real-time quantitative polymerase chain reaction and western blot were employed to determine the expression of hepatic ATP-binding cassette transporter A1 (ABCA1), liver X receptor α (LXRα), and peroxisome proliferator-activated receptor-γ (PPAR-γ). Serum levels of high-density lipoprotein-cholesterol (HDL), low-density lipoprotein-cholesterol (LDL), and total cholesterol (TC) were determined by enzyme-linked immunoassay. Results revealed that PE reversed the formation of atherosclerotic lesion; increased the expression of PPAR-γ, LXRα, and ABCA1; increased the amount of bile flow and total bile acid; reduced the serum level of LDL and TC; and increased the level of HDL. In conclusion, enhancement on bile acid production and consequent cholesterol consumption may partially contribute to the anti-atherosclerotic effects of PE. The reversal of PPARγ-LXRα-ABCA1 signaling pathway may be involved in this process.

Mechanics of the Formation, Interaction, and Evolution of Membrane Tubular Structures.

Membrane nanotubes, also known as membrane tethers, play important functional roles in many cellular processes, such as trafficking and signaling. Although considerable progresses have been made in understanding the physics regulating the mechanical behaviors of individual membrane nanotubes, relatively little is known about the formation of multiple membrane nanotubes due to the rapid occurring process involving strong cooperative effects and complex configurational transitions. By exerting a pair of external extraction upon two separate membrane regions, here, we combine molecular dynamics simulations and theoretical analysis to investigate how the membrane nanotube formation and pulling behaviors are regulated by the separation between the pulling forces and how the membrane protrusions interact with each other. As the force separation increases, different membrane configurations are observed, including an individual tubular protrusion, a relatively less deformed protrusion with two nanotubes on its top forming a V shape, a Y-shaped configuration through nanotube coalescence via a zipper-like mechanism, and two weakly interacting tubular protrusions. The energy profile as a function of the separation is determined. Moreover, the directional flow of lipid molecules accompanying the membrane shape transition is analyzed. Our results provide new, to our knowledge, insights at a molecular level into the interaction between membrane protrusions and help in understanding the formation and evolution of intra- and intercellular membrane tubular networks involved in numerous cell activities.

Directional and Rotational Motions of Nanoparticles on Plasma Membranes as Local Probes of Surface Tension Propagation.

Mechanical heterogeneity is ubiquitous in plasma membranes and of essential importance to cellular functioning. As a feedback of mechanical stimuli, local surface tension can be readily changed and immediately propagated through the membrane, influencing structures and dynamics of both inclusions and membrane-associated proteins. Using the nonequilibrium coarse-grained membrane simulation, here we investigate the inter-related processes of tension propagation, lipid diffusion, and transport of nanoparticles (NPs) adhering on the membrane of constant tension gradient, mimicking that of migrating cells or cells under prolonged stimulation. Our results demonstrate that the lipid bilayer membrane can by itself propagate surface tension in defined rates and pathways to reach a dynamic equilibrium state where surface tension is linearly distributed along the gradient maintained by the directional flow-like motion of lipids. Such lipid flow exerts shearing forces to transport adhesive NPs toward the region of a larger surface tension. Under certain conditions, the shearing force can generate nonzero torques driving the rotational motion of NPs, with the direction of the NP rotation determined by the NP-membrane interaction state as functions of both NP property and local membrane surface tension. Such features endow NPs with promising applications ranging from biosensing to targeted drug delivery.

A multi-channel localized surface plasmon resonance system for absorptiometric determination of abscisic acid by using gold nanoparticles functionalized with a polyadenine-tailed aptamer.

A multi-channel localized surface plasmon resonance system is described for absorptiometric determination of abscisic acid (ABA). The system is making use of gold nanoparticles and consists of a broadband light source, a multi-channel alignment device, and a fiber spectrometer. The method is based on the specific interaction between an ABA-binding aptamer and ABA. This induces the growth of gold nanoparticles (AuNPs) functionalized with a polyadenine-tailed aptamer that act as optical probes. Different concentrations of ABA give rise to varied morphologies of grown AuNPs. This causes a change of absorption spectra which is recorded by the system. ABA can be quantified by measurement of the peak wavelength shifts of grown AuNPs. Under optimized conditions, this method shows a linear relationship in the 1 nM to 10 µM ABA concentration range. The detection limit is 0.51 nM. The sensitivity of the ABA assay is strongly improved compared to the method based on salt-induced AuNP aggregation. This is attributed to the use of a poly-A-tailed aptamer and the catalytic ability of AuNPs. In the actual application, the ABA concentration of ABA in fresh leaves of rice is measured with the maximum relative error of 8.03% in comparison with the ELISA method. Graphical abstractSchematic representation of an absorptiometric approach for determination of abscisic acid based on the growth of polyA-tailed aptamer-AuNPs probes and a multi-channel localized surface plasmon resonance system.

Greedy Mechanism Based Particle Swarm Optimization for Path Planning Problem of an Unmanned Surface Vehicle.

Recently, issues of climate change, environment abnormality, individual requirements, and national defense have caused extensive attention to the commercial, scientific, and military development of unmanned surface vehicles (USVs). In order to design high-quality routes for a multi-sensor integrated USV, this work improves the conventional particle swarm optimization algorithm by introducing the greedy mechanism and the 2-opt operation, based on a combination strategy. First, a greedy black box is established for particle initialization, overcoming the randomness of the conventional method and excluding a great number of infeasible solutions. Then the greedy selection strategy and 2-opt operation are adopted together for local searches, to maintain population diversity and eliminate path crossovers. In addition, Monte-Carlo simulations of eight instances are conducted to compare the improved algorithm with other existing algorithms. The computation results indicate that the improved algorithm has the superior performance, with the shortest route and satisfactory robustness, although a fraction of computing efficiency becomes sacrificed. Moreover, the effectiveness and reliability of the improved method is also verified by its multi-sensor-based application to a USV model in real marine environments.

Application of Improved Particle Swarm Optimization for Navigation of Unmanned Surface Vehicles.

Multi-sensor fusion for unmanned surface vehicles (USVs) is an important issue for autonomous navigation of USVs. In this paper, an improved particle swarm optimization (PSO) is proposed for real-time autonomous navigation of a USV in real maritime environment. To overcome the conventional PSO's inherent shortcomings, such as easy occurrence of premature convergence and human experience-determined parameters, and to enhance the precision and algorithm robustness of the solution, this work proposes three optimization strategies: linearly descending inertia weight, adaptively controlled acceleration coefficients, and random grouping inversion. Their respective or combinational effects on the effectiveness of path planning are investigated by Monte Carlo simulations for five TSPLIB instances and application tests for the navigation of a self-developed unmanned surface vehicle on the basis of multi-sensor data. Comparative results show that the adaptively controlled acceleration coefficients play a substantial role in reducing the path length and the linearly descending inertia weight help improve the algorithm robustness. Meanwhile, the random grouping inversion optimizes the capacity of local search and maintains the population diversity by stochastically dividing the single swarm into several subgroups. Moreover, the PSO combined with all three strategies shows the best performance with the shortest trajectory and the superior robustness, although retaining solution precision and avoiding being trapped in local optima require more time consumption. The experimental results of our USV demonstrate the effectiveness and efficiency of the proposed method for real-time navigation based on multi-sensor fusion.

Stabilizing Effect of Inherent Knots on Proteins Revealed by Molecular Dynamics Simulations.

A growing number of proteins have been identified as knotted in their native structures, with such entangled topological features being expected to play stabilizing roles maintaining both the global fold and the nature of proteins. However, the molecular mechanism underlying the stabilizing effect is ambiguous. Here, we combine unbiased and mechanical atomistic molecular dynamics simulations to investigate how a protein is stabilized by an inherent knot by directly comparing chemical, thermal, and mechanical denaturing properties of two proteins having the same sequence and secondary structures but differing in the presence or absence of an inherent knot. One protein is YbeA from Escherichia coli, containing a deep trefoil knot within the sequence, and the other is the modified protein with the knot of YbeA being removed. Under certain chemical denaturing conditions, the unknotted protein fully unfolds whereas the knotted protein does not, suggesting a higher intrinsic stability for the protein having a knot. Both proteins unfold under enhanced thermal fluctuations but at different rates and with distinct pathways. Opening the hydrophobic core via separation between two α-helices is identified as a crucial step initiating the protein unfolding, which, however, is restrained for the knotted protein by topological and geometrical frustrations. Energy barriers for denaturing the protein are reduced by removing the knot, as evidenced by mechanical unfolding simulations. Finally, yet importantly, no obvious change in size or location of the knot was observed during denaturing processes, indicating that YbeA may remain knotted for a relatively long time during and after denaturation.

Role of Lipid Coating in the Transport of Nanodroplets across the Pulmonary Surfactant Layer Revealed by Molecular Dynamics Simulations.

Hydrophilic drugs can be delivered into lungs via nebulization for both local and systemic therapies. Once inhaled, ultrafine nanodroplets preferentially deposit in the alveolar region, where they first interact with the pulmonary surfactant (PS) layer, with nature of the interaction determining both efficiency of the pulmonary drug delivery and extent of the PS perturbation. Here, we demonstrate by molecular dynamics simulations the transport of nanodroplets across the PS layer being improved by lipid coating. In the absence of lipids, bare nanodroplets deposit at the PS layer to release drugs that can be directly translocated across the PS layer. The translocation is quicker under higher surface tensions but at the cost of opening pores that disrupt the ultrastructure of the PS layer. When the PS layer is compressed to lower surface tensions, the nanodroplet prompts collapse of the PS layer to induce severe PS perturbation. By coating the nanodroplet with lipids, the disturbance of the nanodroplet on the PS layer can be reduced. Moreover, the lipid-coated nanodroplet can be readily wrapped by the PS layer to form vesicular structures, which are expected to fuse with the cell membrane to release drugs into secondary organs. Properties of drug bioavailability, controlled drug release, and enzymatic tolerance in real systems could be improved by lipid coating on nanodroplets. Our results provide useful guidelines for the molecular design of nanodroplets as carriers for the pulmonary drug delivery.

Combined thermodynamic and kinetic analysis of GroEL interacting with CXCR4 transmembrane peptides.

GroEL along with ATP and its co-chaperonin GroES has been demonstrated to significantly enhance the folding of newly translated G-protein-coupled receptors (GPCRs). This work extends the previous studies to explore the guest capture and release processes in GroEL-assisted folding of GPCRs, by the reduced approach of employing CXCR4 transmembrane peptides as model substrates. Each of the CXCR4-derived peptides exhibited high affinity for GroEL with a binding stoichiometry near seven. It is found that the peptides interact with the paired α helices in the apical domain of the chaperonin which are similar with the binding sites of SBP (strongly binding peptide: SWMTTPWGFLHP). Complementary binding study with a single-ring version of GroEL indicates that each of the two chaperonin rings is competent for accommodating all the seven CXCR4 peptides bound to GroEL under saturation condition. Meanwhile, the binding kinetics of CXCR4 peptides with GroEL was also examined; ATP alone, or in combination of GroES evidently promoted the release of the peptide substrates from the chaperonin. The results obtained would be beneficial to understand the thermodynamic and kinetic nature of GroEL-GPCRs interaction which is the central molecular event in the assisted folding process.

Membrane nanotube pearling restricted by confined polymers.

Increasing evidence showed that membrane nanotubes readily undergo pearling in response to external stimuli, while long tubular membrane structures have been observed connecting cells and functioning as channels for intercellular transport, raising a fundamental question of how the stability of membrane nanotubes is maintained in the cellular environment. Here, combining dissipative particle dynamics simulations, free energy calculations, and a force analysis, we propose and demonstrate that nanotube pearling can be restricted by confined polymers, which can be DNA and protein chains transported through the nanotubes, or actin filaments participating in tube formation and elongation. Thermodynamically, nanotube pearling releases the membrane surface energy, but costs bending energies of both the membrane and the confined polymers. Following the mechanism, the pearling of nanotubes confining longer and stiffer polymers is more difficult as it costs larger polymer bending energies. In dynamics, nanotube pearling occurs by repelling polymers from the region of nanotube shrinking to that of swelling. Shorter polymers can be readily repelled owing to the unbalanced force exerted by the shrinking tube region, whereas longer polymers tend to be trapped at the shrinking region to retard the nanotube pearling. Besides the low surface tension maintained by lipid reservoirs kept in living cells, our results supplement the explanation for the stability of membrane nanotubes, and open up a new avenue to manipulate the shape deformation of tubular membrane structures for study of many biological processes.

Regorafenib delays the proliferation of hepatocellular carcinoma by inducing autophagy.

The aim of the present study was to investigate the effects of regorafenib on hepatocellular carcinoma autophagy, thereby supressing the malignancy of HCC. First, HepG2 and Hep3B cell autophagy was investigated using GFP-LC3 transfection after the treatment of regorafenib. Then, the activation of Akt/mTOR signaling was analyzed using western blot. Our data showed that liver cancer cell autophagy was significantly induced by 20 µM regorafenib using GFP-LC3 transfection. Meanwhile, regorafenib-induced cell death could largely be abolished by 3-MA or CQ treatment, suggesting that regorafenib-induced HepG2 cell death was partially dependent on autophagy. Moreover, the activation of Akt/mTOR signaling was inhibited by regorafenib pre-incubation. MTT assay showed the combination use of regorafenib and CDDP led to a stronger growth inhibitory effect on HepG2 and Hep3B cells. In summary, regorafenib may acts an adjunctive therapy for liver cancer patients via modulating autophagy-dependent cell death even when apoptosis resistance is induced in cancer cells.

Interaction pathways between soft lipid nanodiscs and plasma membranes: A molecular modeling study.

Lipid nanodisc, a model membrane platform originally synthesized for study of membrane proteins, has recently been used as the carrier to deliver amphiphilic drugs into target tumor cells. However, the central question of how cells interact with such emerging nanomaterials remains unclear and deserves our research for both improving the delivery efficiency and reducing the side effect. In this work, a binary lipid nanodisc is designed as the minimum model to investigate its interactions with plasma membranes by using the dissipative particle dynamics method. Three typical interaction pathways, including the membrane attachment with lipid domain exchange of nanodiscs, the partial membrane wrapping with nanodisc vesiculation, and the receptor-mediated endocytosis, are discovered. For the first pathway, the boundary normal lipids acting as ligands diffuse along the nanodisc rim to gather at the membrane interface, repelling the central bola lipids to reach a stable membrane attachment. If bola lipids are positioned at the periphery and act as ligands, they diffuse to form a large aggregate being wrapped by the membrane, leaving the normal lipids exposed on the membrane exterior by assembling into a vesicle. Finally, by setting both central normal lipids and boundary bola lipids as ligands, the receptor-mediated endocytosis occurs via both deformation and self-rotation of the nanodiscs. All above pathways for soft lipid nanodiscs are quite different from those for rigid nanoparticles, which may provide useful guidelines for design of soft lipid nanodiscs in widespread biomedical applications.

Lipid extraction mediates aggregation of carbon nanospheres in pulmonary surfactant monolayers.

Increasing evidence indicates that carbon nanoparticles (CNPs), which mainly originate from incomplete combustion of fossil fuels, have an adverse impact on the respiratory system. Recent in vivo experiments have shown that the pulmonary toxicity of CNPs is attributed to their aggregation in pulmonary surfactant monolayers (PSMs) while the underlying mechanism of aggregation remains unclear. Here, by performing coarse grained molecular dynamics simulations, we demonstrate for the first time that the aggregation of carbon nanospheres (CNSs) in PSMs is in fact size-dependent and mediated by lipid extractions. Upon CNS deposition, neighbouring lipid molecules are extracted from PSMs to cover CNSs from the top side. The extracted lipids induce clustering of CNSs to maximize the CNS-lipid interaction, by forming inverse micelles to wrap the aggregated CNSs cooperatively. The formed CNS clusters perturb the molecule structure of the PSM and thus affect its biofunction on respiration. Our simulations show that during the expiration process, CNSs form clusters that perturb the mechanical properties of the PSM in a manner depending on the CNS size. With deep inspiration, a high concentration of large CNSs may induce PSM rupture and thus have a potential impact on its biophysical properties.