A Novel Method for Obtaining Analytical Parameters Based on Double-Flank Measurement.

Double-flank measurement is the most commonly used method for full inspection of mass-produced gears and has high measurement efficiency, but it cannot obtain the analytical parameters and is not helpful enough to evaluate the NVH performance of the gears. Based on the double-flank rolling tester with a new principle, a simulation method for double-flank measurement and a solving method for analytical parameters are proposed. Using the simulation method, the double-flank measurements without random error can be obtained through the collision detection algorithm. The solving method uses the iteration to obtain the minimum rolling length of each position of the tooth surface, then obtains the analytical parameters of the gear. In the experiments, the difference between the profile deviations obtained by the solving method and superimposed in the simulation method is less than 0.03 µm. The experiment results have verified the correctness of the simulation method and the solving method. These methods can greatly improve the value of double-flank measurement.

Self-Calibration Method for Circular Encoders Based on Inertia and a Single Read-Head.

This article proposes a new self-calibration method for circular encoders based on inertia and a single read-head. The velocity curves of the circular encoder are fitted with polynomials and, based on the principle of circle closure and the periodicity of the distribution for angle intervals, the proportionality between the theoretical value and the actual value of each angle interval is obtained. In the experimental system constructed, the feasibility of the proposed method was verified through self-calibration experiments, repeatability experiments, and comparative experiments with the time-measurement dynamic reversal (TDR) method. In addition, this article also proposes an iterative method to improve the self-calibration accuracy. Experimental verification was carried out, and the results show that the new method can effectively compensate for the error of angle measurement in the circular encoder. The peak-to-peak value of the error of angle measurement was reduced from 239.343" to 11.867", and the repeatability of the calibration results of the new method was less than 2.77".

The naturally occurring flavonoid nobiletin reverses methotrexate resistance via inhibition of P-glycoprotein synthesis.

Methotrexate (MTX) is the first-line treatment for rheumatoid arthritis (RA). However, after long-term treatment, some patients develop resistance. P-glycoprotein (P-gp), as an indispensable drug transporter, is essential for mediating this MTX resistance. In addition, nobiletin (NOB), a naturally occurring polymethoxylated flavonoid, has also been shown to reverse P-gp-mediated MTX resistance in RA groups; however, the precise role of NOB in this process is still unclear. Here, we administered MTX and NOB alone or in combination to collagen II-induced arthritic (CIA) mice and evaluated disease severity using the arthritis index, synovial histopathological changes, immunohistochemistry, and P-gp expression. In addition, we used conventional RNA-seq to identify targets and possible pathways through which NOB reverses MTX-induced drug resistance. We found that NOB in combination with MTX could enhance its performance in synovial tissue and decrease P-gp expression in CIA mice compared to MTX treatment alone. In vitro, in MTX-resistant fibroblast-like synoviocytes from CIA cells (CIA-FLS/MTX), we show that NOB treatment downregulated the PI3K/AKT/HIF-1α pathway, thereby reducing the synthesis of the P-gp protein. In addition, NOB significantly inhibited glycolysis and metabolic activity of CIA-FLS/MTX cells, which could reduce the production of ATP and block P-gp, ultimately decreasing the efflux of MTX and maintaining its anti-RA effects. In conclusion, this study shows that NOB overcomes MTX resistance in CIA-FLS/MTX cells through the PI3K/AKT/HIF-1α pathway, simultaneously influencing metabolic processes and inhibiting P-gp-induced drug efflux.

A Normalized Absolute Values Adaptive Evaluation Function of Image Clarity.

The clarity evaluation function plays a vital role in the autofocus technique. The accuracy and efficiency of the image clarity evaluation function directly affects the accuracy of autofocus and the speed of focusing. However, classical clarity function values are sensitive to changes in background brightness and changes in object contour length. This paper proposes a normalized absolute values adaptive (NAVA) evaluation function of image clarity. It can eliminate the influence of changes in background brightness and the length of the measured object contour on the image clarity function value. To verify the effectiveness of the NAVA function, several experiments were conducted under conditions of virtual master gear images and actual captured images. For actual captured images, the variation of the evaluation results of the NAVA function is far less than the corresponding variation of the classic clarity function. Compared with classical clarity evaluation functions, the NAVA function can provide normalized absolute clarity values. The correlations between the NAVA function results of image clarity and both the contour length and background brightness of the tested object are weak. The use of the NAVA function in automatic and manual focusing systems can further improve focusing efficiency.

Study on the Measurability of Gear Analytical Parameters in Double-Flank Measurement.

Double-flank measurement is the most commonly used full inspection method on the shop floor. However, the double-flank measurement method cannot measure analytical parameters such as pitch deviations and profile deviations, and this limitation is a pain point in the field of gear measurement. This paper studies the measurability of the analytical parameters of gears based on the results of double-flank measurement, proposes the definition of measurable area, and gives the relationship between the size of the measurable area and the number of teeth and the pressure angle and the gear error. Digital simulation methods were used to conduct measurement experiments on gear analytical parameters. In the experiments, the measurability of the analytical parameters of gears with various typical profile deviations in the double-flank measurement process was verified and analyzed. The test results show that not all profile deviations are unmeasurable in the process of double-flank measurement, but there exists a profile region in which the analytical parameters of the gear can be measured accurately. The size of the measurable area of the profile is mainly determined by the number of teeth and pressure angle of the gear, while the pitch deviations are always measurable under normal conditions.

Effect of charge on protein preferred orientation at the air-water interface in cryo-electron microscopy.

The air-water interface (AWI) tends to adsorb proteins and frequently causes preferred orientation problems in cryo-electron microscopy (cryo-EM). Here, we examined cryo-EM data from protein samples frozen with different detergents and found that both anionic and cationic detergents promoted binding of proteins to the AWI. By contrast, some of the nonionic and zwitterionic detergents tended to prevent proteins from attaching to the AWI. The protein orientation distributions with different anionic detergents were similar and resembled that obtained without detergent. By contrast, cationic detergents gave distinct orientation distributions. Our results indicate that proteins adsorb to charged interface and the negative charge of the AWI plays an important role in adsorbing proteins in the conventional cryo-EM sample preparation. According to these findings, a new method was developed by adding anionic detergent at a concentration between 0.002% and 0.005%. Using this method, the protein particles exhibited a more evenly distributed orientations and still adsorbed to the AWI enabling them embedding in a thin layer of ice with high concentration, which will benefit the cryo-EM structural determination.

Hierarchical drug release designed Au @PDA-PEG-MTX NPs for targeted delivery to breast cancer with combined photothermal-chemotherapy.

Breast cancer (BC) is the most frequently diagnosed cancer with a low survival rate and one of the major causes of cancer-related death. Methotrexate (MTX) is an anti-tumor drug used in the treatment of BC. Poor dispersion in water and toxic side effects limit its clinical application. Gold nanoparticles (AuNPs), owing to their specific structures and unique biological and physiochemical properties, have emerged as potential vehicles for tumor targeting, bioimaging and cancer therapy. An innovative nano drug-loading system (Au @PDA-PEG-MTX NPs) was prepared for targeted treatment of BC. Au @PDA-PEG-MTX NPs under near infra-red region (NIR) irradiation showed effective photothermal therapy against MDA-MB-231 human BC cells growth in vitro by inducing apoptosis through triggering reactive oxygen species (ROS) overproduction and generating excessive heat. In vivo studies revealed deep penetration ability of Au @PDA-PEG-MTX NPs under NIR irradiation to find application in cancer-targeted fluorescence imaging, and exhibited effective photothermal therapy against BC xenograft growth by inducing apoptosis. Histopathological analysis, cellular uptake, cytotoxicity assay, and apoptosis experiments indicated that Au @PDA-PEG-MTX NPs possessed a good therapeutic effect with high biocompatibility and fewer side effects. This Au NPs drug-loading system achieved specific targeting of MTX to BC cells by surface functionalisation, fluorescence imaging under laser irradiation, combined photothermal-chemotherapy, and pH- and NIR- triggered hierarchical drug release.

Recent advances of nanotechnology-based tumor vessel-targeting strategies.

Tumor vessels can provide oxygen and nutrition for solid tumor tissue, create abnormal tumor microenvironment (TME), and play a vital role in the development, immune escape, metastasis and drug resistance of tumor. Tumor vessel-targeting therapy has become an important and promising direction in anti-tumor therapy, with the development of five anti-tumor therapeutic strategies, including vascular disruption, anti-angiogenesis, vascular blockade, vascular normalization and breaking immunosuppressive TME. However, the insufficient drug accumulation and severe side effects of vessel-targeting drugs limit their development in clinical application. Nanotechnology offers an excellent platform with flexible modified surface that can precisely deliver diverse cargoes, optimize efficacy, reduce side effects, and realize the combined therapy. Various nanomedicines (NMs) have been developed to target abnormal tumor vessels and specific TME to achieve more efficient vessel-targeting therapy. The article reviews tumor vascular abnormalities and the resulting abnormal microenvironment, the application of NMs in the tumor vessel-targeting strategies, and how NMs can improve these strategies and achieve multi-strategies combination to maximize anti-tumor effects.

High-quality, high-throughput cryo-electron microscopy data collection via beam tilt and astigmatism-free beam-image shift.

The throughput of cryo-electron microscopy (cryo-EM) can be improved by employing a procedure that collects beam-image shift data. However, this procedure inadvertently induces a beam tilt, thus decreasing the resolution of the reconstruction. Here, we report an automatic calibration procedure for correcting the beam tilt and a large unexpected astigmatism in the beam-image shift data collection. In this procedure, the changes of the beam tilt and the astigmatism against the beam shift are measured and calibrated. The beam tilt and the astigmatism are corrected by changing the setting of the microscope using predicted values from the calibration. Using our corrected beam-image shift data collection, we found that the resolution remained identical as long as the distance of the beam shift was below 10 µm. The image throughput increases by ∼80%, with image quality improving by reducing the residual stage drift, thus benefiting the high resolution cryo-EM structure determination. Such a calibration procedure takes about 3 h and can be applied to different microscopes.

Research on Path Planning Model Based on Short-Term Traffic Flow Prediction in Intelligent Transportation System.

Vehicle driving path planning is an important information service in intelligent transportation systems. As an important basis for path planning optimization, the travel time prediction method has attracted much attention. However, traffic flow has features of high nonlinearity, time-varying, and uncertainty, which makes it hard for prediction method with single feature to meet the accuracy demand of intelligent transportation system in big data environment. In this paper, the historical vehicle Global Positioning System (GPS) information data is used to establish the traffic prediction model. Firstly, the Clustering in QUEst (CLIQUE)-based clustering algorithm V-CLIQUE is proposed to analyze the historical vehicle GPS data. Secondly, an artificial neural network (ANN)-based prediction model is proposed. Finally, the ANN-based weighted shortest path algorithm, A-Dijkstra, is proposed. We used mean absolute percentage error (MAPE) to evaluate the predictive model and compare it with the predicted results of Average and support regression vector (SRV). Experiments show that the improved ANN path planning model we proposed can accurately predict real-time traffic status at the given location. It has less relative error and saves time for users' travel while saving social resources.

Virus structures revealed by advanced cryoelectron microscopy methods.

Before the resolution revolution, cryoelectron microscopy (cryo-EM) single-particle analysis (SPA) already achieved resolutions beyond 4 Å for certain icosahedral viruses, enabling ab initio atomic model building of these viruses. As the only samples that achieved such high resolution at that time, cryo-EM method development was closely intertwined with the improvement of reconstructions of symmetrical viruses. Viral morphology exhibits significant diversity, ranging from small to large, uniform to non-uniform, and from containing single symmetry to multiple symmetries. Furthermore, viruses undergo conformational changes during their life cycle. Several methods, such as asymmetric reconstruction, Ewald sphere correction, cryoelectron tomography (cryo-ET), and sub-tomogram averaging (STA), have been developed and applied to determine virus structures in vivo and in vitro. This review outlines current advanced cryo-EM methods for high-resolution structure determination of viruses and summarizes accomplishments obtained with these approaches. Moreover, persisting challenges in comprehending virus structures are discussed and we propose potential solutions.

The reduction of FIB damage on cryo-lamella by lowering energy of ion beam revealed by a quantitative analysis.

Focused ion beam (FIB) is widely used for thinning frozen cells to produce lamellae for cryo-electron microscopy imaging and for protein structures study in vivo. However, FIB damages the lamellae and a quantitative experimental analysis of the damage is lacking. We used a 30-keV gallium FIB to prepare lamellae of a highly concentrated icosahedral virus sample. The viruses were grouped according to their distance from the surface of lamellae and reconstructed. Damage to the approximately 20-nm-thick outermost lamella surface was similar to that from exposure to 16 e-/Å2 in a 300-kV cryo-electron microscope at high-resolution range. The damage was negligible at a depth beyond 50 nm, which was reduced to 30 nm if 8-keV Ga+ was used during polishing. We designed extra steps in the reconstruction refinement to maximize undamaged signals and increase the resolution. The results demonstrated that low-energy beam polishing was essential for high-quality thinner lamellae.

Near-atomic architecture of Singapore grouper iridovirus and implications for giant virus assembly.

Singapore grouper iridovirus (SGIV), one of the nucleocytoviricota viruses (NCVs), is a highly pathogenic iridovirid. SGIV infection results in massive economic losses to the aquaculture industry and significantly threatens global biodiversity. In recent years, high morbidity and mortality in aquatic animals have been caused by iridovirid infections worldwide. Effective control and prevention strategies are urgently needed. Here, we present a near-atomic architecture of the SGIV capsid and identify eight types of capsid proteins. The viral inner membrane-integrated anchor protein colocalizes with the endoplasmic reticulum (ER), supporting the hypothesis that the biogenesis of the inner membrane is associated with the ER. Additionally, immunofluorescence assays indicate minor capsid proteins (mCPs) could form various building blocks with major capsid proteins (MCPs) before the formation of a viral factory (VF). These results expand our understanding of the capsid assembly of NCVs and provide more targets for vaccine and drug design to fight iridovirid infections.

Passivating Lead Halide Perovskites Using Pyridinium Salts with Superhalogen Atoms.

Passivating lead halide perovskites using pyridinium salts has attracted enormous attention, but the excellent surface passivation of the halide perovskites has not been achieved by using only a pyridinium salt until now. Herein, passivating the (001) planes of the cubic CsPbI3, CH3NH3PbI3, and NH2CHNH2PbI3 perovskites using the pyridinium salts of C5NH6X (X = Cl, Br, I, PF6, ClO4, or BF4) is systematically studied by high-throughput first-principle calculations and ab initio molecular dynamics simulations. The results show that the excellent surface passivation of the three perovskites is achieved by the pyridinium salt of C5NH6BF4 (i.e., shallow level, negative formation energy, unchanged band-edge construction, and stable dynamics property are obtained for the three passivated perovskites), which strongly imply that their devices can show excellent performances, such as long-term stability, low ion migration, and high efficiency. However, the C5NH6ClO4 and C5NH6PF6 pyridinium salts are only profitable for passivating the (001) PbI2 plane of the three perovskites, and other C5NH6X pyridinium salts have adverse effects.

An electron counting algorithm improves imaging of proteins with low-acceleration-voltage cryo-electron microscope.

Relative to the 300-kV accelerating field, electrons accelerated under lower voltages are potentially scattered more strongly. Lowering the accelerate voltage has been suggested to enhance the signal-to-noise ratio (SNR) of cryo-electron microscopy (cryo-EM) images of small-molecular-weight proteins (<100 kD). However, the detection efficient of current Direct Detection Devices (DDDs) and temporal coherence of cryo-EM decrease at lower voltage, leading to loss of SNR. Here, we present an electron counting algorithm to improve the detection of low-energy electrons. The counting algorithm increased the SNR of 120-kV and 200-kV cryo-EM image from a Falcon III camera by 8%, 20% at half the Nyquist frequency and 21%, 80% at Nyquist frequency, respectively, resulting in a considerable improvement in resolution of 3D reconstructions. Our results indicate that with further improved temporal coherence and a dedicated designed camera, a 120-kV cryo-electron microscope has potential to match the 300-kV microscope at imaging small proteins.

Hyaluronic Acid Hydrogels Hybridized With Au-Triptolide Nanoparticles for Intraarticular Targeted Multi-Therapy of Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory disease, characterized by synovial inflammation in multiple joints. Triptolide (TP) is a disease-modifying anti-rheumatic drug (DMARD) highly effective in patients with RA and has anti-inflammatory properties. However, its clinical application has been limited owing to practical disadvantages. In the present study, hyaluronic acid (HA) hydrogel-loaded RGD-attached gold nanoparticles (AuNPs) containing TP were synthesized to alleviate the toxicity and increase therapeutic specificity. The hydrogels can be applied for targeted photothermal-chemo treatment and in vivo imaging of RA. Hydrogel systems with tyramine-modified HA (TA-HA) conjugates have been applied to artificial tissue models as surrogates of cartilage to investigate drug transport and release properties. After degradation of HA chains, heat was locally generated at the inflammation region site due to near-infrared resonance (NIR) irradiation of AuNPs, and TP was released from nanoparticles, delivering heat and drug to the inflamed joints simultaneously. RA can be penetrated with NIR light. Intraarticular administration of the hydrogels containing low dosage of TP with NIR irradiation improved the inflamed conditions in mice with collagen-induced arthritis (CIA). Additionally, in vitro experiments were applied to deeply verify the antirheumatic mechanisms of TP-PLGA-Au@RGD/HA hydrogels. TP-PLGA-Au@RGD/HA hydrogel treatment significantly reduced the migratory and invasive capacities of RA fibroblast-like synoviocytes (RA-FLS) in vitro, through the decrease of phosphorylation of mTOR and its substrates, p70S6K1, thus inhibiting the mTOR pathway.

Low-cooling-rate freezing in biomolecular cryo-electron microscopy for recovery of initial frames.

When biological samples are first exposed to electrons in cryo-electron microcopy (cryo-EM), proteins exhibit a rapid 'burst' phase of beam-induced motion that cannot be corrected with software. This lowers the quality of the initial frames, which are the least damaged by the electrons. Hence, they are commonly excluded or down-weighted during data processing, reducing the undamaged signal and the resolution in the reconstruction. By decreasing the cooling rate during sample preparation, either with a cooling-rate gradient or by increasing the freezing temperature, we show that the quality of the initial frames for various protein and virus samples can be recovered. Incorporation of the initial frames in the reconstruction increases the resolution by an amount equivalent to using ~60% more data. Moreover, these frames preserve the high-quality cryo-EM densities of radiation-sensitive residues, which is often damaged or very weak in canonical three-dimensional reconstruction. The improved freezing conditions can be easily achieved using existing devices and enhance the overall quality of cryo-EM structures.

miR-18a-5p Targets FBP1 to Promote Proliferation, Migration, and Invasion of Liver Cancer Cells and Inhibit Cell Apoptosis.

Liver cancer is one of the most aggressive malignant tumors. It is significant to understand the molecular mechanism of liver cancer cells to develop new treatment plans. Studies have identified that FBP1 serves as a cancer inhibitor gene. To research the effect mechanism of FBP1 in liver cancer cells, bioinformatics analysis was performed to study its expression in liver cancer tissue. Survival analysis was also performed. Moreover, starBase database was applied to predict upstream regulatory genes of FBP1. Dual-luciferase assay was performed to testify their targeted relationship. The mRNA and protein expression levels of FBP1 in liver cancer cells were detected by qRT-PCR and western blot, respectively. Cell viability was analyzed by CCK-8 assay. The migratory and invasive abilities of cells were analyzed by Transwell assay. The apoptosis of liver cancer cells was detected by flow cytometry. The results showed that the expression of FBP1 was downregulated in liver cancer tissue and cells. FBP1 low expression was correlated with the poor prognosis of patients. miR-18a-5p could inhibit FBP1 expression. Overexpression of FBP1 could inhibit the progression of liver cancer cells and promote cell apoptosis. Overexpressing miR-18a-5p could promote the progression of liver cancer cells and inhibit cell apoptosis. However, overexpressing FBP1 simultaneously could reverse the effect. miR-18a-5p and FBP1 are expected to be candidates for liver cancer treatment.

Screening Hub Genes of Hepatocellular Carcinoma Based on Public Databases.

Tumor recurrence and metastasis often occur in HCC patients after surgery, and the prognosis is not optimistic. Hence, searching effective biomarkers for prognosis of is of great importance. Firstly, HCC-related data was acquired from the TCGA and GEO databases. Based on GEO data, 256 differentially expressed genes (DEGs) were obtained firstly. Subsequently, to clarify function of DEGs, clusterProfiler package was used to conduct functional enrichment analyses on DEGs. Protein-protein interaction (PPI) network analysis screened 20 key genes. The key genes were filtered via GEPIA database, by which 11 hub genes (F9, CYP3A4, ASPM, AURKA, CDC20, CDCA5, NCAP, PRC1, PTTG1, TOP2A, and KIFC1) were screened out. Then, univariate Cox analysis was applied to construct a prognostic model, followed by a prediction performance validation. With the risk score calculated by the model and common clinical features, univariate and multivariate analyses were carried out to assess whether the prognostic model could be used independently for prognostic prediction. In conclusion, the current study screened HCC prognostic gene signature based on public databases.

Nab-paclitaxel promotes the cancer-immunity cycle as a potential immunomodulator.

Paclitaxel is a widely used anti-tumor chemotherapeutic drug. Solvent-based paclitaxel causes bone marrow suppression, allergic reactions, neurotoxicity and systemic toxicity, which are associated with non-specific cytotoxicity and side effects of fat-soluble solvents. Studies have explored various new nano-drug strategies of paclitaxel, including nanoparticle albumin-bound paclitaxel (nab-paclitaxel) to improve the water solubility and safety of paclitaxel. Nab-paclitaxel is a targeted solvent-free formulation that inhibits microtubule depolymerization to anticancer. It is easily taken up by tumor and immune cells owing to the nano-scaled size and superior biocompatibility. The internalized nab-paclitaxel exhibits significant immunostimulatory activities to promote cancer-immunity cycle. The aim of this study was to explore the synergistic effect of nab-paclitaxel in tumor antigen presentation, T cell activation, reversing the immunosuppressive pattern of tumor microenvironment (TME), and the synergistic effect with cytotoxic lymphocytes (CTLs) in clearance of tumor cells. The effects of nab-paclitaxel on modulation of cancer-immunity cycle, provides potential avenues for combined therapeutic rationale to improve efficacy of immunotherapy.