The Bcl-2/xL inhibitor ABT-263 increases the stability of Mcl-1 mRNA and protein in hepatocellular carcinoma cells

Background Hepatocellular carcinoma (HCC) is one of the major causes of mortality. ABT-263 is a newly synthesized, orally available Bcl-2/xL inhibitor that shows promising efficacy in HCC therapy. ABT-263 inhibits the anti-apoptotic activity of Bcl-2 and Bcl-xL, but not Mcl-1. Previous reports have shown that ABT-263 upregulates Mcl-1 in various cancer cells, which contributes to ABT-263 resistance in cancer therapy. However, the associated mechanisms are not well known. Methods Western blot, RNAi and CCK-8 assays were used to investigate the relationship between Mcl-1 upregulation and ABT-263 sensitivity in HCC cells. Real-time PCR and Western blot were used to detect Mcl-1 mRNA and protein levels. Luciferase reporter assay and RNA synthesis inhibition assay were adopted to analyze the mechanism of Mcl-1 mRNA upregulation. Western blot and the inhibition assays for protein synthesis and proteasome were used to explore the mechanisms of ABT-263-enhanced Mcl-1 protein stability. Trypan blue exclusion assay and flow cytometry were used to examine cell death and apoptosis. Results ABT-263 upregulated Mcl-1 mRNA and protein levels in HCC cells, which contributes to ABT-263 resistance. ABT-263 increased the mRNA level of Mcl-1 in HCC cells by enhancing the mRNA stability without influencing its transcription. Furthermore, ABT-263 increased the protein stability of Mcl-1 through promoting ERK- and JNK-induced phosphorylation of Mcl-1Thr163 and increasing the Akt-mediated inactivation of GSK-3β. Additionally, the inhibitors of ERK, JNK or Akt sensitized ABT-263-induced apoptosis in HCC cells. Conclusions ABT-263 increases Mcl-1 stability at both mRNA and protein levels in HCC cells. Inhibition of ERK, JNK or Akt activity sensitizes ABT-263-induced apoptosis. This study may provide novel insights into the Bcl-2-targeted cancer therapeutics.This study was supported in part by Chongqing Natural Science Foundation (cstc2011BB5030 and 2013jjB10015), the National Natural Science Foundation of China (31201068, 81273226 and 81228005) and the Scientific Funds of Third Military Medical University (2011XHG02 and 2012XZH01)

Effect of Deep Cryogenic Treatment on Microstructure and Properties of Sintered Fe–Co–Cu-Based Diamond Composites

Metal-matrix-impregnated diamond composites are used for fabricating many kinds of diamond tools. In the efforts to satisfy the increasing engineering requirements, researchers have brought more attention to find novel methods of enhancing the performance of impregnated diamond composites. In this study, deep cryogenic treatment was applied to Fe–Co–Cu-based diamond composites to improve their performance. Relative density, hardness, bending strength, and grinding ratio of matrix and diamond composite samples were measured by a series of tests. Meanwhile, the fracture morphologies of all samples after the bending strength test were observed and analyzed by scanning electron microscope. The results showed that the hardness and bending strength of matrix increased slightly after deep cryogenic treatment. The grinding ratio of impregnated diamond composites exhibited a great increase by 32.9% as a result of deep cryogenic treatment. The strengthening mechanism was analyzed in detail. The Fe–Co–Cu-based impregnated composites subjected to deep cryogenic treatment for 1 h exhibited the best overall performance

Coupling analysis of contra-rotating fan interstage pressure pulsation and blade vibration based on wavelet reconstruction.

In recent years, the flow characteristics research of the interstage region in counter-rotating axial fans in terms of fluid dynamics has attracted much attention. Especially, the dynamic relationship between interstage pressure pulsation and blade vibration in counter-rotating axial fans has not yet been clarified. This paper performs the signal processing method of wavelet decomposition and reconstruction in time-frequency analysis process. Under different flow conditions, weak-coupling numerical simulation program is employed to analyze the fluid-structure coupling interaction between interstage pressure pulsations and blade vibrations in counter-rotating axial fans. The results indicate that the fluid-structure coupling interaction field in the interstage of counter-rotating axial fans mainly excites the first-order vibration of the second-stage blade. At the same time, the consistency between the pulsation frequency and the vibration frequency of the airflow reflects the good coupling property. Two stage blades cut the airflow to cause field changes and airflow pulsation, and then, airflow pulsation causes blades deformation and produces vibrations of the same frequency at the blade. The deformation of the blades, in turn, causes the flow field changes. Rotating stall, vortex movement and breakdown produced low-frequency airflow pulsation and vortex vibration of the blade

Photoelectrochemical sensor based on Bi2(Te1-xSex)3 for the sensitive detection of the tetracycline hydrochloride

Tetracycline hydrochloride (TC-HCl) as an antibiotic is widely used in our daily life, but it has posed potential threats to the ecological environment for flowing into the natural environment. To address this issue, The photoelectrochemical (PEC) sensor is first developed for the detection of TC-HCl by comparing and analyzing ternary Bi2(Te1-xSex)3 nanomaterials prepared through different ratios of hydrothermal methods. Specifically, the Bi2(Te1-xSex)3/Indium tin oxide (ITO) electrode can interact with TC-HCl rapidly and selectively which results in a significant reduction of the photocurrent signal. By regulating the ratio of x, it is found that Bi2Te2.85Se0.15/ITO have the best stability and the fastest response time in the detection of TC-HCl. The response/recovery speed of the sensor to TC-HCl is fast to 0.23 s, a low detection limit of 0.08 pmol/l (S/N = 3) and a linear detection fit of up to R2 = 0.992. Moreover, the sensor has good stability and specificity for TC-HCL detection, providing a new perspective for the development of PEC sensors

Low-dose imaging denoising with one pair of noisy images

Low-dose imaging techniques have many important applications in diverse fields, from biological engineering to materials science. Samples can be protected from phototoxicity or radiation-induced damage using low-dose illumination. However, imaging under a low-dose condition is dominated by Poisson noise and additive Gaussian noise, which seriously affects the imaging quality, such as signal-to-noise ratio, contrast, and resolution. In this work, we demonstrate a low-dose imaging denoising method that incorporates the noise statistical model into a deep neural network. One pair of noisy images is used instead of clear target labels and the parameters of the network are optimized by the noise statistical model. The proposed method is evaluated using simulation data of the optical microscope, and scanning transmission electron microscope under different low-dose illumination conditions. In order to capture two noisy measurements of the same information in a dynamic process, we built an optical microscope that is capable of capturing a pair of images with independent and identically distributed noises in one shot. A biological dynamic process under low-dose condition imaging is performed and reconstructed with the proposed method. We experimentally demonstrate that the proposed method is effective on an optical microscope, fluorescence microscope, and scanning transmission electron microscope, and show that the reconstructed images are improved in terms of signal-to-noise ratio and spatial resolution. We believe that the proposed method could be applied to a wide range of low-dose imaging systems from biological to material science.Published versionThis work was funded by National Key Research and Development Program of China (2021YFB3602604); National Natural Science Foundation of China (61975205,62075221, 62131011); Fusion Foundation of Research and Education of CAS; University of Chinese Academy of Sciences; Fundamental Research Funds for the Central Universities; Funded Project of Hebei Province Innovation Capability Improvement Plan, China (20540302D)

Corrosion Evaluation of Pure Mg Coated by Fluorination in 0.1 M Fluoride Electrolyte

In the ongoing research on the application of biodegradable materials, surface treatment of is considered to be a relatively effective solution to the excessive degradation rates of Mg alloys. In this study, to further optimize the proven effective surface coatings of fluoride, a low-voltage preparation fluorination method was used to achieve coating effectiveness under safer conditions. Optical observation, scanning electron microscopy (SEM), X-ray diffraction (XRD), energy-dispersive spectroscopy (EDS), and potential dynamic polarization (PDP) experiments were used for the analysis and evaluation. The coating characteristics of the MgF2 coatings treated in the 10–90 V voltage range, including the structure, chemical conformation, and electrochemical corrosion assessment, were fully defined. The anodic fluoridation results showed that a pore structure of 1–14 μm thickness was formed on the Mg alloy substrate, and the coating was composed of Mg fluoride. The results of immersion corrosion and electrochemical corrosion experiments showed that compared with pure Mg, anodic fluorinated samples below 40 V exhibited better corrosion resistance, the prepared MgF2 coating was more uniform, and the surface mostly exhibited point corrosion. When the voltage reached or exceeded 60 V, the prepared coating exhibited poor corrosion resistance, fracture, and protrusions. After corrosion, it mostly exhibited surface corrosion. The results indicate that idealized coatings can be obtained at relatively low and safe voltage ranges. This finding may enable more economical, environmentally friendly, and safe preparation of coatings

In Vivo Corrosion Behavior of Biodegradable Magnesium Alloy by MAF Treatment

Coating treatment plays an irreplaceable role in propelling the clinical application of magnesium alloys. This experiment was designed in order to observe the anticorrosion behavior of magnesium fluoride coating in rats. The MgF2 layer was prepared on the surface of AZ31 magnesium alloy in saturated NH4HF2 solution by microarc fluorination (MAF) at 190 V. The cross-sectional SEM, EDS, and XRD analysis indicated that the alloy surface was covered with MgF2. Meanwhile, SEM observation was used to compare the magnesium alloy samples before and after treatment, and it was found that the samples after coating were flatter and smoother. Two sets of experiments were carried out with the subjects, 6-week-old male rats. So that the untreated AZ31 samples and the microarc fluorinated AZ31 samples could be buried under the muscle layer individually. The volume changes and surface morphology of the corroded samples were monitored dynamically using micro-CT over a 16-week period in vivo. Comparison of results between the two sets of samples presented that the corrosion of the microarc fluoridated samples was much slower than that of the untreated ones. The MAF coating was shown to be effective in controlling the corrosion rate and progression of the magnesium alloy

AKT-mediated phosphorylation of ATG4B impairs mitochondrial activity and enhances the Warburg effect in hepatocellular carcinoma cells

<p>Phosphorylation is a major type of post-translational modification, which can influence the cellular physiological function. ATG4B, a key macroautophagy/autophagy-related protein, has a potential effect on the survival of tumor cells. However, the role of ATG4B phosphorylation in cancers is still unknown. In this study, we identified a novel phosphorylation site at Ser34 of ATG4B induced by AKT in HCC cells. The phosphorylation of ATG4B at Ser34 had little effect on autophagic flux, but promoted the Warburg effect including the increase of L-lactate production and glucose consumption, and the decrease of oxygen consumption in HCC cells. The Ser34 phosphorylation of ATG4B also contributed to the impairment of mitochondrial activity including the inhibition of F₁Fo-ATP synthase activity and the elevation of mitochondrial ROS in HCC cells. Moreover, the phosphorylation of ATG4B at Ser34 enhanced its mitochondrial location and the subsequent colocalization with F₁Fo-ATP synthase in HCC cells. Furthermore, recombinant human ATG4B protein suppressed the activity of F₁Fo-ATP synthase in MgATP submitochondrial particles from patient-derived HCC tissues in vitro. In brief, our results demonstrate for the first time that the phosphorylation of ATG4B at Ser34 participates in the metabolic reprogramming of HCC cells via repressing mitochondrial function, which possibly results from the Ser34 phosphorylation-induced mitochondrial enrichment of ATG4B and the subsequent inhibition of F₁Fo-ATP synthase activity. Our findings reveal a noncanonical working pattern of ATG4B under pathological conditions, which may provide a scientific basis for developing novel strategies for HCC treatment by targeting ATG4B and its Ser34 phosphorylation.</p