Background::Radiation (IR)-induced DNA damage triggers cell cycle arrest and has a suppressive effect on the tumor microenvironment (TME). Wee1, a cell cycle regulator, can eliminate G2/M arrest by phosphorylating cyclin-dependent kinase 1 (CDK1). Meanwhile, programed death-1/programed death ligand-1 (PD-1/PDL-1) blockade is closely related to TME. This study aims to investigate the effects and mechanisms of Wee1 inhibitor AZD1775 and anti-PD-1 antibody (anti-PD-1 Ab) on radiosensitization of hepatoma.Methods::The anti-tumor activity of AZD1775 and IR was determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay on human and mouse hepatoma cells HepG2, Hepa1-6, and H22. The anti-hepatoma mechanism of AZD1775 and IR revealed by flow cytometry and Western blot in vitro. A hepatoma subcutaneous xenograft mice model was constructed on Balb/c mice, which were divided into control group, IR group, AZD1775 group, IR + AZD1775 group, IR + anti-PD-1 Ab group, and the IR + AZD1775 + anti-PD-1 Ab group. Cytotoxic CD8 + T cells in TME were analyzed by flow cytometry. Results::Combining IR with AZD1775 synergistically reduced the viability of hepatoma cells in vitro. AZD1775 exhibited antitumor effects by decreasing CDK1 phosphorylation to reverse the IR-induced G2/M arrest and increasing IR-induced DNA damage. AZD1775 treatment also reduced the proportion of PD-1 +/CD8 + T cells in the spleen of hepatoma subcutaneous xenograft mice. Further studies revealed that AZD1775 and anti-PD-1 Ab could enhance the radiosensitivity of hepatoma by enhancing the levels of interferon γ (IFNγ) + or Ki67 + CD8 T cells and decreasing the levels of CD8 + Tregs cells in the tumor and spleen of the hepatoma mice model, indicating that the improvement of TME was manifested by increasing the cytotoxic factor IFNγ expression, enhancing CD8 + T cells proliferation, and weakening CD8 + T cells depletion. Conclusions::This work suggests that AZD1775 and anti-PD-1 Ab synergistically sensitize hepatoma to radiotherapy by enhancing IR-induced DNA damage and improving cytotoxic CD8 + T cells in TME.

Objective:To investigate the impact of the deep learning reconstruction algorithm TrueFidelity TM for Gemstone Spectral Imaging (TF-GSI) and the adaptive statistical iterative reconstruction algorithm (ASiR-V, hereinafter referred to as ASiR-V) based on phantom and animal models on the image quality of dual-energy CT images. Methods:GE Revolution Apex CT was used to scan the ACR 464 phantom and a mouse model of gastric cancer with lymph node metastasis ( n=16). TF-GSI and ASiR-V were separately used to reconstruct middle and high-grade images (TF-GSI-M, TF-GSI-H, ASiR-V-50%, and ASiR-V-100%) on the phantom and mouse based on virtual monoenergetic images at 70 keV. The task transfer function (TTF) of bone and acrylic, image noise power spectrum (NPS), and detectability index (d′) of the phantom images were evaluated. One-way ANOVA analysis was used to compare the image noise, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) for brain and liver on images of mice. The consistency of the two reconstruction-algorithm images (TF-GSI-H and ASiR-V100%) in the detection of small lesions by two radiologists (A and B) was evaluated using kappa test. Results:In terms of the phantom, the TF-GSI-H group had the best performance in TTF, NPS, and d′. Compared to ASiR-V-100%, the TTF50% of bone and acrylic in the TF-GSI-H group increased by 2.4% and 8.9%, respectively; the NPS peak decreased by 54.1%, compared to ASiR-V-100%; the d′ of bone and acrylic in the TF-GSI-H group relative to ASiR-V-100% increased by 52.7% and 59.5%, respectively. The TF-GSI group had reduced image noise compared to the ASiR-V group, and both SNR and CNR of the two tissues increased, but the differences between the groups were not statistically significant (all P>0.05). The two reconstruction-algorithm images showed good consistency in image evaluation by the two radiologists (A, Kappa=0.875, P<0.001; B, Kappa=0.625, P=0.012). In terms of the detection of micro-metastases in mice, the TF-GSI group outperformed the ASiR-V group (average accuracy: 83.5% vs 71.9%; average sensitivity: 77.8% vs 61.2%; average specificity: 85.7% vs 85.7%). Conclusion:Compared with iterative reconstruction algorithm, the DLIR algorithm showed improved spatial resolution, reduced image noise, and enabled detectability of micro-lesion for images from dual-energy CT.

Objective::This study aimed to evaluate the major adverse cardiovascular and cerebrovascular events (MACCEs) and overall safety profile associated with iodixanol in Chinese patients undergoing percutaneous coronary intervention (PCI).Methods::Patients at 30 centers in China registered in the OpenClinic v3.6 database from October 30, 2013, to October 7, 2015, were included in the study. The primary endpoint was in-hospital MACCEs including target lesion revascularization (TLR), stroke, stent thrombosis, cardiac death, and PCI-related myocardial infarction (MI) within 72 h post-PCI. Secondary endpoints were MACCEs from 72 h to 30 d post-PCI and other safety events within 30 d post-PCI.Results::A total of 3,042 patients were enrolled. The incidence of MACCEs within 72 h post-PCI was 2.33% ( n = 71), including cardiac death (0.03%, n = 1) and PCI-related MI (2.30%, n = 70). The incidence of MACCEs from 72 h to 30 d post-PCI was 0.16% ( n = 5), including cardiac death (0.10%, n = 3), PCI-related MI (0.03%, n = 1), and TLR for stent thrombosis (0.03%, n = 1). The incidence of composite angiographic or procedural complications was 2.86% ( n = 87); 233 (7.86%) patients had results suggesting contrast-induced acute kidney injury. Conclusions::These findings indicate that the use of iodixanol in Chinese patients undergoing PCI is associated with a low incidence of MACCEs, confirming its safety in this population.

Objective::This study aimed to evaluate the major adverse cardiovascular and cerebrovascular events (MACCEs) and overall safety profile associated with iodixanol in Chinese patients undergoing percutaneous coronary intervention (PCI).Methods::Patients at 30 centers in China registered in the OpenClinic v3.6 database from October 30, 2013, to October 7, 2015, were included in the study. The primary endpoint was in-hospital MACCEs including target lesion revascularization (TLR), stroke, stent thrombosis, cardiac death, and PCI-related myocardial infarction (MI) within 72 h post-PCI. Secondary endpoints were MACCEs from 72 h to 30 d post-PCI and other safety events within 30 d post-PCI.Results::A total of 3,042 patients were enrolled. The incidence of MACCEs within 72 h post-PCI was 2.33% ( n = 71), including cardiac death (0.03%, n = 1) and PCI-related MI (2.30%, n = 70). The incidence of MACCEs from 72 h to 30 d post-PCI was 0.16% ( n = 5), including cardiac death (0.10%, n = 3), PCI-related MI (0.03%, n = 1), and TLR for stent thrombosis (0.03%, n = 1). The incidence of composite angiographic or procedural complications was 2.86% ( n = 87); 233 (7.86%) patients had results suggesting contrast-induced acute kidney injury. Conclusions::These findings indicate that the use of iodixanol in Chinese patients undergoing PCI is associated with a low incidence of MACCEs, confirming its safety in this population.

Extracellular traps released by neutrophils (neutrophil extracellular traps, NETs) are a double-edged sword, and understanding the mechanism of NET formation is of great significance for disease treatment. However, the short lifespan, the large individual differences, and the inability to perform gene editing render it difficult to decipher NET formation using neutrophils. It is necessary to find a model cell to replace neutrophils to study the mechanism of NET formation. In this study, we used different concentrations (0, 0.1, 1, and 10 μmol/L) of all-trans retinoic acid (ATRA) to differentiate HL-60 cells for different days (1, 3, 5, and 7 days). By detecting the cell viability and nuclear morphology of cells, we confirmed that HL-60 cells were differentiated to neutrophil-like cells (dHL-60) after treated with ATRA for at least 5 days. Using immunofluorescence staining to detect the formation of NETs, we demonstrated that dHL-60 cells differentiated for 5 days with 1 μmol/L ATRA could generate NETs comparable to those produced by neutrophils upon phorbol 12-myristate 13-acetate (PMA) stimulation, without histone H3 citrullination. Furthermore, the formation of NETs by dHL-60 cells were NADPH-dependent and PAD4-independent, consistent with neutrophils. Taken together, these observations suggest that dHL-60 cells differentiated with 1 μmol/L ATRA for 5 days can be used as a model cell for neutrophils to study the mechanism of NET formation.
Humans ; Extracellular Traps ; Tetradecanoylphorbol Acetate/pharmacology* ; Neutrophils ; HL-60 Cells ; Tretinoin/pharmacology*

Background::Ubiquitination plays an essential role in many biological processes, including viral infection, and can be reversed by deubiquitinating enzymes (DUBs). Although some studies discovered that DUBs inhibit or enhance viral infection by various mechanisms, there is lack of information on the role of DUBs in virus regulation, which needs to be further investigated.Methods::Immunoblotting, real-time polymerase chain reaction, in vivo/ in vitro deubiquitination, protein immunoprecipitation, immunofluorescence, and co-localization biological techniques were employed to examine the effect of ubiquitin-specific protease 3 (USP3) on APOBEC3G (A3G) stability and human immunodeficiency virus (HIV) replication. To analyse the relationship between USP3 and HIV disease progression, we recruited 20 HIV-infected patients to detect the levels of USP3 and A3G in peripheral blood and analysed their correlation with CD4 + T-cell counts. Correlation was estimated by Pearson correlation coefficients (for parametric data). Results::The results demonstrated that USP3 specifically inhibits HIV-1 replication in an A3G-dependent manner. Further investigation found that USP3 stabilized 90% to 95% of A3G expression by deubiquitinating Vif-mediated polyubiquitination and blocking its degradation in an enzyme-dependent manner. It also enhances the A3G messenger RNA (mRNA) level by binding to A3G mRNA and stabilizing it in an enzyme-independent manner. Moreover, USP3 expression was positively correlated with A3G expression ( r= 0.5110) and CD4 + T-cell counts ( r= 0.5083) in HIV-1-infected patients. Conclusions::USP3 restricts HIV-1 viral infections by increasing the expression of the antiviral factor A3G. Therefore, USP3 may be an important target for drug development and serve as a novel therapeutic strategy against viral infections.

Neutrophil extracellular traps (NETs) play an important role in the formation of immunothrombosis. However, how vascular endothelial cells mediate the formation of NETs has not been fully understood. We stimulated neutrophils firmly attached on the endothelial cell surface intercellular adhesion molecule-1 (ICAM-1) with lipopolysaccharide (LPS) or phorbol-12-myristate-13-acetate (PMA) for 4 h, then labeled NETs-DNA with Sytox green dye and the formation of NETs was observed by fluorescent microscopy. The area and fluorescence intensity of NETs-DNA were analyzed to quantify the formation of NETs. The results showed that both PMA and LPS were able to induce firmly adhered neutrophils on ICAM-1 to produce NETs. NETs induced by PMA were independent of neither β2 integrin lymphocyte function-associated antigen-1 (LFA-1) nor macrophage antigen complex-1 (Mac-1). In contrast, LPS-stimulated NETs were mediated by Mac-1 integrin, but not by LFA-1. After inhibition of actin filaments or Talin-1, the formation of NETs irrespective of the stimulus was significantly reduced. This study reveals the mechanism of the direct interaction between neutrophils and endothelial cells to produce NETs under inflammatory conditions, providing a new theoretical basis for the treatment of related diseases and the development of new drugs.
Cytoskeletal Proteins ; Endothelial Cells ; Extracellular Traps ; Integrins ; Intercellular Adhesion Molecule-1 ; Lipopolysaccharides/pharmacology* ; Macrophages ; Neutrophils

PURPOSE: To investigate the effects of hyperbaric oxygen (HBO) pretreatment on cognitive decline and neuronal damage in an Alzheimer’s disease (AD) rat model. MATERIALS AND METHODS: Rats were divided into three groups: normal saline (NS), AD, and HBO+AD. In the AD group, amyloid β peptide (Aβ)₁₋₄₀ was injected into the hippocampal CA1 region of the brain. NS rats received NS injection. In the HBO+AD group, rats received 5 days of daily HBO therapy following Aβ₁₋₄₀ injection. Learning and memory capabilities were examined using the Morris water maze task. Neuronal damage and astrocyte activation were evaluated by hematoxylin-eosin staining and immunohistochemistry, respectively. Dendritic spine density was determined by Golgi-Cox staining. Tumor necrosis factor-α, interleukin-1β, and interleukin-10 production was assessed by enzyme-linked immunosorbent assay. Neuron apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling. Protein expression was examined by western blotting. RESULTS: Learning and memory dysfunction was ameliorated in the HBO+AD group, as shown by significantly lower swimming distances and escape latency, compared to the AD group. Lower rates of neuronal damage, astrocyte activation, dendritic spine loss, and hippocampal neuron apoptosis were seen in the HBO+AD than in the AD group. A lower rate of hippocampal p38 mitogen-activated protein kinase (MAPK) phosphorylation was observed in the HBO+AD than in the AD group. CONCLUSION: HBO pretreatment improves cognition and reduces hippocampal damage via p38 MAPK in AD rats.
Alzheimer Disease/*therapy ; Amyloid beta-Peptides/*administration & dosage ; Animals ; Apoptosis ; *Cognition/drug effects ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Hippocampus/*enzymology ; *Hyperbaric Oxygenation ; In Situ Nick-End Labeling ; Interleukin-10/biosynthesis ; Interleukin-1beta/biosynthesis ; Learning/drug effects ; Male ; Memory/drug effects ; Neurons ; Peptide Fragments/*administration & dosage ; Rats ; Rats, Sprague-Dawley ; Sodium Chloride/administration & dosage ; Tumor Necrosis Factor-alpha/biosynthesis ; p38 Mitogen-Activated Protein Kinases/*metabolism

Objective To explore the neurasthenia situation of the population attending the postgraduate exam in medical class and its influencing factors .Methods The random cluster sampling method was adopted .The influencing factors were per‐formed the χ2 test and Logistic regression analysis .Results According to the inclusion criteria and exclusion criteria ,36 positive ca‐ses were screened out ,accounting for 8 .89% of participants ,which was higher than the value of the general population ;the statisti‐cally significant differences were noted in the factors of the confidence of attending the postgraduate exam ,stress ,adjusting ability , interpersonal situation ,reading duration ,sleep ,nutrition and dietary (P<0 .05) .Conclusion Attending the postgraduate exam such highly intensitive mental activity is the important reason causing neurasthenia among the population attending the postgraduate ex‐am ,which mainly display in the aspects of stress size ,bearing ability facing stress and regulation .

Objective To investigate the effect of intrathecal nerve growth factor (NGF) on lidocaineinduced neurotoxicity to the spinal cord in rats.Methods Thirty healthy male Sprague-Dawley rats,aged 8-10 weeks,weighing 250-300 g,were randomly divided into 5 groups (n =6 each) using a random number table:control group (group C),sham operation group (group S),lidocaine-induced neurotoxicity group (group L),normal saline group (group NS) and NGF group (group NGF).A catheter was inserted at L4.5 interspace into the epidural space in S,L,NS,and NGF groups.One day after surgery,20％ lidocaine 20 μl was injected intrathecally.At 24 h after lidocaine injection,normal saline 20 μl and NGF 10 μg (20 μd) were injected intrathecally in NS and NGF groups,respectively,once a day for 7 consecutive days.The tail flick latency (TFL) to a thermal nociceptive stimulus was measured on 1,3,5 and 7 days after lidocaine injection.The animals were sacrificed after the behavioral test was completed at 7 days after lidocaine injection,and the lumbar segments of the spinal cord were removed to detect the neuronal apoptosis (using flow cytometry) and caspase-3 mRNA expression (by RT-PCR).Results Compared with group C,the TFL was significantly prolonged at 1-7 days after lidocaine injection in L and NS groups and at 1-5 days after lidocaine injection in group NGF,the apoptosis rate was increased in L,NS and NGF groups,and caspase-3 mRNA expression was up-regulated in L and NS groups,and no significant change was found in the parameters mentioned above in group S.Compared with group L,the TFL was significantly shortened at 5 and 7 days after lidocaine injection,the apoptosis rate was decreased,and caspase3 mRNA expression was down-regulated in NGF group,and no significant change was found in the parameters mentioned above in group NS.Conclusion Intrathecal NGF can reduce lidocaine-induced neurotoxicity to the spinal cord in rats and inhibition of caspase-3 mRNA expression is involved in the mechanism.