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Abnormal fluctuations in the mold can easily lead to slag entrainment during the thin slab continuous casting process. It is difficult to guarantee the uniform growth of the shell solidification of the slab. The quality of the slab is seriously affected. In this study, the data on thin slab mold level fluctuation were collected from a complete casting sequence of low-carbon steel in a Chinese plant. The discrete wavelet transform (DWT) method is used to decompose the mold level fluctuation and different process parameters. The frequency composition of abnormal fluctuations was clarified, and the causes of the abnormal fluctuations were revealed. The results show that the frequency ranges of abnormal fluctuations are concentrated in 0.3125–0.625 Hz and 0.625–1.25 Hz, and the low set mold level is where the maximum amplitude occurs. Within the time-frequency range, changes in parameters such as electromagnetic braking, mold oscillation, casting speed, stopper rod, and tundish weight cannot be completely matched to abnormal fluctuations collected by the eddy current sensor (ECS). It was found that the abnormal fluctuations in the ECS were that its equipment was disturbed through the analysis of the fluctuations collected by the radioactive source detection (RSD). The degree of disturbance is related to the height of the set mold level. The maximum fluctuation difference exceeded 6 mm between different collection methods when the set mold level was low. In this plant, the position of the ECS is to avoid the area of violent fluctuations of the surface molten steel and be moved appropriately towards the submerged entry nozzle (SEN) to reduce disturbance. When the current ECS position is unchanged, the mold level is set at approximately 305–309 mm, and the casting speed is matched to 4.8 m/min, which minimizes the disturbance with the ECS measurement.

Abstract Background Harsh parenting has been recognized as a risk factor for adolescent anxiety; however, the underlying mechanisms of this relationship remain unclear, and it is unknown whether this relationship is influenced by different parental roles and living arrangements. This study aimed to investigate the mediating mechanisms between harsh parenting and adolescent anxiety symptoms using a multiple mediation model and to further compare specific roles of harsh parenting and distinguish between the living arrangements. Methods A total of 3505 adolescents completed this survey, and 3295 adolescents (54.7% girls, Mage = 14.97 years) were included in the study. Participants completed self-assessments measuring harsh parenting, self-efficacy, school connectedness, Internet addiction, sleep problems, and anxiety. They were categorized into three groups based on living arrangements: living with both parents, only with the mother, or only with the father. Results Correlational analyses revealed that both maternal and paternal harsh parenting were associated with increased anxiety symptoms. Structural equation modeling (SEM) mediation analyses and multigroup analyses showed that the independent mediating effects of school connectedness, Internet addiction, and sleep problems, as well as the sequential mediating pathways involving self-efficacy → school connectedness, self-efficacy → Internet addiction, and self-efficacy → sleep problems, vary across the adolescents’ living arrangements in the association between maternal and paternal harsh parenting and adolescent anxiety symptoms. Conclusions This study elucidated the mechanisms linking harsh parenting to adolescent anxiety symptoms and validated the effects of different parental roles and living arrangements. The findings provide important insights for developing targeted interventions to address anxiety symptoms in adolescents exposed to harsh parenting.

Abstract Immune checkpoint inhibitors (ICIs) immunotherapy facilitates new approaches to achieve precision cancer treatment. A growing number of patients with non-small cell lung cancer (NSCLC) have benefited from treatment with neoadjuvant ICIs combined with chemotherapy. However, the mechanisms and associations between the therapeutic efficacy of neoadjuvant pembrolizumab and chemotherapy (NAPC) and macrophage subsets are still unclear. We performed single-cell RNA sequencing (scRNA-seq) and identified a novel FABP4+C1q+ macrophage subtype, which exhibited stronger proinflammatory cytokine production and phagocytic ability. This subtype was found to be more abundant in tumor tissues and lymph nodes of major pathological response (MPR) patients compared to non-MPR patients, and was associated with a good efficacy of NAPC. Multiplex fluorescent immunohistochemical (mIHC) staining was subsequently used to verify our findings. Further mechanistic studies indicated that FABP4 and C1q regulate the expression of proinflammatory cytokines synergistically. In addition, FABP4 and C1q promote fatty acid synthesis, enhance anti-apoptosis ability and phagocytic ability of macrophage via the interaction of AMPK/JAK/STAT axis. This study provides novel insights into the underlying mechanisms and predictive biomarkers of NAPC. Our findings contribute to improving the prognosis of patients with NSCLC by potentially guiding more precise patient selection and treatment strategies. Novelty & Impact Statements We identified a group of macrophages (FABP4+C1q+ macrophages) related to the therapeutic efficacy of neoadjuvant chemoimmunotherapy. FABP4+C1q+ macrophages highly expressed proinflammatory cytokines-related genes and had a strong cytokine production and phagocytic ability. We believe that our study provides a novel insight into the synergistic mechanism of neoadjuvant ICI combined with chemotherapy and may lead to improved clinical outcomes in patients with NSCLC in the future.

Abstract There is a growing interest in the creation of engineered condensates formed via liquid–liquid phase separation (LLPS) to exert precise cellular control in prokaryotes. However, de novo design of cellular condensates to control metabolic flux or protein translation remains a challenge. Here, we present a synthetic condensate platform, generated through the incorporation of artificial, disordered proteins to realize specific functions in Bacillus subtilis. To achieve this, the “stacking blocks” strategy is developed to rationally design a series of LLPS-promoting proteins for programming condensates. Through the targeted recruitment of biomolecules, our investigation demonstrates that cellular condensates effectively sequester biosynthetic pathways. We successfully harness this capability to enhance the biosynthesis of 2’-fucosyllactose by 123.3%. Furthermore, we find that condensates can enhance the translation specificity of tailored enzyme fourfold, and can increase N-acetylmannosamine titer by 75.0%. Collectively, these results lay the foundation for the design of engineered condensates endowed with multifunctional capacities.

Abstract Objective The aim of our study is to find a better way to identify a group of papillary thyroid carcinoma (PTC) with more aggressive behaviors and to provide a prediction model for lymph node metastasis to assist in clinic practice. Methods Targeted sequencing of DNA/RNA was used to detect genetic alterations. Gene expression level was measured by quantitative real-time PCR, western blotting or immunohistochemistry. CCK8, transwell assay and flow cytometry were used to investigate the effects of concomitant gene alterations in PTC. LASSO-logistics regression algorithm was used to construct a nomogram model integrating radiomic features, mutated genes and clinical characteristics. Results 172 high-risk variants and 7 fusion types were detected. The mutation frequencies in BRAF, TERT, RET, ATM and GGT1 were significantly higher in cancer tissues than benign nodules. Gene fusions were detected in 16 samples (2 at the DNA level and 14 at the RNA level). ATM mutation (ATM MUT) was frequently accompanied by BRAF MUT, TERT MUT or gene fusions. ATM MUT alone or ATM co-mutations were significantly positively correlated with lymph node metastasis. Accordingly, ATM knock-down PTC cells bearing BRAF V600E, KRAS G12R or CCDC6-RET had higher proliferative ability and more aggressive potency than cells without ATM knock-down in vitro. Furthermore, combining gene alterations and clinical features significantly improved the predictive efficacy for lymph node metastasis of radiomic features, from 71.5 to 87.0%. Conclusions Targeted sequencing of comprehensive genetic alterations in PTC has high prognostic value. These alterations, in combination with clinical and radiomic features, may aid in predicting invasive PTC with higher accuracy.

PurposeCoinfections or consecutive infections of Mucor and Aspergillus are exceedingly uncommon in children, we report the case to offer the valuable experience for colleagues facing similar situations.Case reportThis report documents the first recorded case of successful treatment for pulmonary mixed infection in a diabetic girl. Initially, the patient underwent treatment based on voriconazole, but the infection continued to deteriorate. Subsequently, bronchoalveolar lavage fluid culture and metagenomic next-generation sequencing (mNGS) were conducted, leading to a clear diagnosis of simultaneous infection by Aspergillus fumigatus and Rhizopus microsporus. Susceptibility testing revealed fungal resistance to voriconazole. Therefore, a combined treatment regimen of AmB liposomes and isavuconazole effectively eradicated the fungal infection.ConclusionThis case underscores the importance of early and precise identification of fungal pathogens, determination of effective antifungal medications, and timely implementation of well-planned therapeutic strategies. Furthermore, we comprehensively reviewed 10 cases of pulmonary mixed infections involving Mucor and Aspergillus, summarizing their characteristics and identifying commonalities.

Halophytes face continuous challenges from environmental salt conditions throughout their lifecycle, and adaptive strategies are essential for their growth and survival. Hordeum brevisubulatum, a halophytic forage grass, relies on efficient K+ uptake for salt tolerance, as demonstrated by the inducible expression of HbHAK2 (high affinity K+ transporter 2). Using genome walking, we isolated a unique 744-bp HbHAK2 promoter, distinct from the upstream sequences of HvHAK2 in barley. Transgenic Arabidopsis plants carrying the GUS gene, driven by the HbHAK2 promoter, exhibited higher activity in tissues and cells exhibiting polar growth, including root hairs, lateral root initiation sites, trichomes, and shoot apical meristems (SAM), particularly under various stresses compared to control conditions. In both Arabidopsis and Brachypodium distachyon, the expression of HbHAK2, driven by its promoter (proHbHAK2) improved K+ uptake, promoted a root architecture characterized by increased lateral branching, and enhanced salt tolerance by augmenting the root system (including lateral roots and root-hair development and growth) under salt stress. Therefore, variation in the HbHAK2 promoter represents a specific strategy for halophytic H. brevisubulatum to cope with salt stress, which holds potential for improving salt tolerance in crop breeding programs.

Cognitive dysfunction is a core symptom common in psychiatric disorders including depression that is primarily managed by antidepressants lacking efficacy in improving cognition. In this study, we report a novel dual serotonin transporter and voltage-gated potassium Kv7/KCNQ/M-channel inhibitor D01 (a 2-methyl-3-aryloxy-3-heteroarylpropylamines derivative) that exhibits both anti-depression effects and improvements in cognition. D01 inhibits serotonin transporters (Ki = 30.1 ± 6.9 nmol/L) and M channels (IC50 = 10.1 ± 2.4 μmol/L). D01 also reduces the immobility duration in the mouse FST and TST assays in a dose-dependent manner without a stimulatory effect on locomotion. Intragastric administrations of D01 (20 and 40 mg/kg) can significantly shorten the immobility time in a mouse model of chronic restraint stress (CRS)-induced depression-like behavior. Additionally, D01 dose-dependently improves the cognitive deficit induced by CRS in Morris water maze test and increases the exploration time with novel objects in normal or scopolamine-induced cognitive deficits in mice, but not fluoxetine. Furthermore, D01 reverses the long-term potentiation (LTP) inhibition induced by scopolamine. Taken together, our findings demonstrate that D01, a dual-target serotonin reuptake and M channel inhibitor, is highly effective in the treatment-resistant depression and cognitive deficits, thus holding potential for development as therapy of depression with cognitive deficits.

In the contemporary era, there is a heightened awareness of the significance of a spiritual life, and travel is a popular choice for many. The Internet is driving rapid diversification in people's travel choices. At the same time, the traditional tourism industry's Tourism Management (TM) model, which has been in place for decades, is no longer able to satisfy people's diversified choices for tourism. This is due to a number of factors, including the trust crisis caused by asymmetric information, the outdated nature of the TM model, and the insecure personal information that is shared between intermediaries. This paper discusses the application of sustainable blockchain technology in tourism management and sets up a tourism management system based on blockchain and the Internet of Things. It takes 20 scenic spots as examples to study the application of the TM model. This paper evaluates the TM model from four key aspects: tourist satisfaction, tourism infrastructure completeness, tourism consumption level and tourism service content richness. It compares the results with those of the traditional TM system. The experimental results are clear: tourist satisfaction at scenic spots 5, 10 and 15 in the sustainable blockchain TM mode is 68 %, 87 % and 71 % respectively, which is higher than that in the traditional TM mode. In the context of sustainable blockchain, the TM mode is the optimal approach for serving tourists and enhancing their travel experience. This study also assists Chinese tourism business operators in recognizing the potential impact of blockchain technology on the development of the tourism industry. They can then formulate strategies at an early stage to cope with the information technology changes in the era of the digital economy.

Abstract Objective Immunotherapy, specifically immune checkpoint inhibitors (ICIs), has revolutionized cancer treatment. However, it can also cause immune‐related adverse events (irAEs). This study aimed to develop a clinically practical animal model of irAEs using BALB/c mice. Methods Subcutaneous tumors of mouse breast cancer 4T1 cells were generated in inbred BALB/c mice. The mice were treated with programmed death‐1 (PD‐1) and cytotoxic t‐lymphocyte antigen 4 (CTLA‐4) inhibitors once every 3 days for five consecutive administration cycles. Changes in tumor volume and body weight were recorded. Lung computed tomography (CT) scans were conducted. The liver, lungs, heart, and colon tissues of the mice were stained with hematoxylin–eosin (H&E) staining to observe inflammatory infiltration and were scored. Serum samples were collected, and enzyme‐linked immunosorbent assay (ELISA) was used to detect the levels of ferritin, glutamic‐pyruvic transaminase (ALT), tumor necrosis factor‐α (TNF‐α), interferon‐gamma (IFN‐γ), and interleukin‐6 (IL‐6). Mouse liver and lung cell suspensions were prepared, and changes in macrophages, T cells, myeloid‐derived suppressor cells (MDSCs), and regulatory (Treg) cells were detected by flow cytometry. Results Mice treated with PD‐1 and CTLA‐4 inhibitors showed significant reductions in tumor volume and body weight. The tissue inflammatory scores in the experimental group were significantly higher than those in the control group. Lung CT scans of mice in the experimental group showed obvious inflammatory spots. Serum levels of ferritin, IL‐6, TNF‐α, IFN‐γ, and ALT were significantly elevated in the experimental group. Flow cytometry analysis revealed a substantial increase in CD3+T cells, Treg cells, and macrophages in the liver and lung tissues of mice in the experimental group compared with the control group, and the change trend of MDSCs was opposite. Conclusions The irAE‐related animal model was successfully established in BALB/c mice using a combination of PD‐1 and CTLA‐4 inhibitors through multiple administrations with clinical translational value and practical. This model offers valuable insights into irAE mechanisms for further investigation.

Abstract Biosynthesis is the application of enzymes in microbial cell factories and has emerged as a promising alternative to chemical synthesis. However, natural enzymes with limited catalytic performance often need to be engineered to meet specific needs through a time‐consuming trial‐and‐error process. This study presents a quantum mechanics (QM)‐incorporated design–build–test–learn (DBTL) framework to rationally design phosphatase BT4131, an enzyme with an ambiguous substrate spectrum involved in N‐acetylglucosamine (GlcNAc) biosynthesis. First, mutant M1 (L129Q) is designed using force field‐based methods, resulting in a 1.4‐fold increase in substrate preference (kcat/Km) toward GlcNAc‐6‐phosphate (GlcNAc6P). QM calculations indicate that the shift in substrate preference is caused by a 13.59 kcal mol−1 reduction in activation energy. Furthermore, an iterative computer‐aided design is conducted to stabilize the transition state. As a result, mutant M4 (I49Q/L129Q/G172L) with a 9.5‐fold increase in kcat‐GlcNAc6P/Km‐GlcNAc6P and a 59% decrease in kcat‐Glc6P/Km‐Glc6P is highly desirable compared to the wild type in the GlcNAc‐producing chassis. The GlcNAc titer increases to 217.3 g L−1 with a yield of 0.597 g (g glucose)−1 in a 50‐L bioreactor, representing the highest reported level. Collectively, this DBTL framework provides an easy yet fascinating approach to the rational design of enzymes for industrially viable biocatalysts.

Abstract Background High-performance computing plays a pivotal role in computer-aided drug design, a field that holds significant promise in pharmaceutical research. The prediction of drug–target affinity (DTA) is a crucial stage in this process, potentially accelerating drug development through rapid and extensive preliminary compound screening, while also minimizing resource utilization and costs. Recently, the incorporation of deep learning into DTA prediction and the enhancement of its accuracy have emerged as key areas of interest in the research community. Drugs and targets can be characterized through various methods, including structure-based, sequence-based, and graph-based representations. Despite the progress in structure and sequence-based techniques, they tend to provide limited feature information. Conversely, graph-based approaches have risen to prominence, attracting considerable attention for their comprehensive data representation capabilities. Recent studies have focused on constructing protein and drug molecular graphs using sequences and SMILES, subsequently deriving representations through graph neural networks. However, these graph-based approaches are limited by the use of a fixed adjacent matrix of protein and drug molecular graphs for graph convolution. This limitation restricts the learning of comprehensive feature representations from intricate compound and protein structures, consequently impeding the full potential of graph-based feature representation in DTA prediction. This, in turn, significantly impacts the models’ generalization capabilities in the complex realm of drug discovery. Results To tackle these challenges, we introduce GLCN-DTA, a model specifically designed for proficiency in DTA tasks. GLCN-DTA innovatively integrates a graph learning module into the existing graph architecture. This module is designed to learn a soft adjacent matrix, which effectively and efficiently refines the contextual structure of protein and drug molecular graphs. This advancement allows for learning richer structural information from protein and drug molecular graphs via graph convolution, specifically tailored for DTA tasks, compared to the conventional fixed adjacent matrix approach. A series of experiments have been conducted to validate the efficacy of the proposed GLCN-DTA method across diverse scenarios. The results demonstrate that GLCN-DTA possesses advantages in terms of robustness and high accuracy. Conclusions The proposed GLCN-DTA model enhances DTA prediction performance by introducing a novel framework that synergizes graph learning operations with graph convolution operations, thereby achieving richer representations. GLCN-DTA does not distinguish between different protein classifications, including structurally ordered and intrinsically disordered proteins, focusing instead on improving feature representation. Therefore, its applicability scope may be more effective in scenarios involving structurally ordered proteins, while potentially being limited in contexts with intrinsically disordered proteins.

Aiming at the problem of system controller performance failure caused by improperly setting the value of each weighting coefficient of the model predictive control (MPC), a fractional-order MPC strategy with Takagi–Sugeno fuzzy optimization (T–SFO MPC) is proposed for a vehicle active suspension system. Firstly, the fractional-order model predictive control framework for active suspension systems is designed based on a 1/4 vehicle model. Then, we analyze the influence of different weighting coefficients on the suspension performance and introduce the Takagi–Sugeno fuzzy optimization theory to adaptively adjust the weighting coefficients of the fractional-order MPC controller. Finally, the system responses of the T–SFO MPC, traditional MPC, linear quadratic regulator (LQR), and passive suspension control are numerically analyzed under various road conditions. Simulation results show that suspension response with the T–SFO MPC is significantly improved compared with passive suspension control, traditional MPC control, and LQR control, and the weight coefficients of the T–SFO MPC can be adaptively adjusted according to the dynamic changes of suspension response. Compared with passive suspension, the root mean square (RMS) value of the vertical acceleration of the T–SFO MPC under various roads decreased by a maximum of 37.97%, and the RMS value of suspension dynamic deflection and tire dynamic load decreased by a maximum of 32.94% and 37.8%, respectively. These results validate that the proposed control method can achieve coordinated optimization of vehicle comfort and handling stability.

The enhancement of the electromagnetic interference shielding efficiency (EMI SE) for conductive polymer composites (CPCs) has garnered increasing attention. The shielding performance is influenced by conductivity, which is dependent on the establishment of effective conductive pathways. In this review, Schelkunoff’s theory on outlining the mechanism of electromagnetic interference shielding was briefly described. Based on the mechanism, factors that influenced the electrical percolation threshold of CPCs were presented and three main kinds of efficient methods were discussed for establishing conductive pathways. Furthermore, examples were explored that highlighted the critical importance of such conductive pathways in attaining optimal shielding performance. Finally, we outlined the prospects for the future direction for advancing CPCs towards a balance of enhanced EMI SE and cost–performance.

ObjectiveThe objective of this study is to elucidate the influence of MCU on the clinical pathological features of GC patients, to investigate the function and mechanism of the mitochondrial calcium uptake transporter MCU in the initiation and progression of GC, and to explore its impact on the metabolic pathways and biosynthesis of mitochondria. The ultimate goal is to identify novel targets and strategies for the clinical management of GC patients.MethodsTumor and adjacent tissue specimens were obtained from 205 patients with gastric cancer, and immunohistochemical tests were performed to assess the expression of MCU and its correlation with clinical pathological characteristics and prognosis. Data from TCGA, GTEx and GEO databases were retrieved for gastric cancer patients, and bioinformatics analysis was utilized to investigate the association between MCU expression and clinical pathological features. Furthermore, we conducted an in-depth analysis of the role of MCU in GC patients. We investigated the correlation between MCU expression in GC and its impact on mitochondrial function, metabolism, biosynthesis, and immune cells. Additionally, we studied the proteins or molecules that interact with MCUResultsOur research revealed high expression of MCU in the GC tissues. This high expression was associated with poorer T and N staging, and indicated a worse disease-free survival period. MCU expression was positively correlated with mitochondrial function, mitochondrial metabolism, nucleotide, amino acid, and fatty acid synthesis metabolism, and negatively correlated with nicotinate and nicotinamide metabolism. Furthermore, the MCU also regulates the function of the mitochondrial oxidative respiratory chain. The MCU influences the immune cells of GC patients and regulates ROS generation, cell proliferation, apoptosis, and resistance to platinum-based drugs in gastric cancer cells.ConclusionHigh expression of MCU in GC indicates poorer clinical outcomes. The expression of the MCU are affected through impacts the function of mitochondria, energy metabolism, and cellular biosynthesis in gastric cancer cells, thereby influencing the growth and metastasis of gastric cancer cells. Therefore, the mitochondrial changes regulated by MCU could be a new focus for research and treatment of GC.

BackgroundImmune checkpoint therapy, involving the programmed cell death 1 (PD-1) monoclonal antibody, has revolutionized the treatment of cancer. Tertiary lymphatic structure (TLS) serves as an immune indicator to predict the efficacy of PD-1 antibody therapy. However, there is no clear result whether the distribution, quantity, and maturity of TLS can be effective indicators for predicting the clinical efficacy of anti-PD1 immunotherapy in patients with colorectal cancer (CRC).MethodsFifty-seven patients who underwent surgical resection and thirty-nine patients who received anti-PD-1 immunotherapy were enrolled in this retrospective study. Immunohistochemical staining and multiple fluorescence immunohistochemistry were used to evaluate the mismatch repair (MMR) subtypes and TLS distribution, quantity, and maturity, respectively.ResultsA comprehensive patient score system was built based on TLS quantity and maturity. We found that the proportion of patients with score >1 was much higher in the deficient mismatch repair(dMMR) group than in the proficient mismatch repair(pMMR) group, and this difference was mainly due to intratumoral TLS. Patient score, based on the TLS evaluation of whole tumor, peritumor, or intratumor, was used to evaluate the efficacy of anti-PD1 immunotherapy. Based only on the intratumor TLS evaluation, the proportion of patients with a score >1 was higher in the response (PR + CR) group than in the non-response (PD) group. Multivariate analysis revealed that patient scores were positively correlated with the clinical efficacy of immunotherapy. Further analysis of immune-related progression-free survival was performed in patients with CRC who received anti-PD-1 immunotherapy. Patients with score >1 based on the intratumor TLS evaluation had significantly better survival.ConclusionsThese results suggest that the patient score based on intratumor TLS evaluation may be a good immune predictive indicator for PD-1 antibody therapy in patients with CRC.

Summary: Although the impact of sleep loss on social behaviors has been widely observed in recent years, the mechanisms underpinning these impacts remain unclear. In this study, we explored the detrimental effects of sleep deprivation on reciprocity behavior as well as its underlying psychological and neuroimaging mechanisms by combining sleep manipulation, an interpersonal interactive game, computational modeling and neuroimaging. Our results suggested that after sleep deprivation, individuals showed reduced reciprocity behavior, mainly due to their reduced weights on communal concern when making social decisions. At neural level, we demonstrated that sleep deprivation’s effects were observed in the precuneus (hyperactivity) and temporoparietal junction, dorsal lateral prefrontal cortex (DLPFC) (both hypoactivity), and reduced reciprocity was also accounted for by increased precuneus-thalamus connectivity and DLPFC-thalamus connectivity. Our findings contributed to the understanding of the psychological and neuroimaging bases underlying the deleterious impact of sleep deprivation on social behaviors.

This study aims to provide a high-value and environmentally friendly method for the application of coal-based solid waste. Modified fly ash/polyurethane (MFA/PU) and modified coal gangue powder/polyurethane (MCG/PU) composites were prepared by adding different contents of MFA and MCG (10%, 20%, 30%, 40%). At the filler content of 30%, the compressive strengths of MFA/PU and MCG/PU are 84.1 MPa and 46.3 MPa, respectively, likely due to an improvement in interface compatibility, as indicated by scanning electron microscopy (SEM). The MFA/PU and MCG/PU composites present their highest limiting oxygen index (LOI) values of 29% and 23.5%, respectively, when their filler content is 30%. MFA has advantages in improving the LOIs of composites. Cone calorimetry (CCT) and SEM demonstrate that the two composites exhibit similar condensed-phase flame-retardant behaviors during combustion, which releases CO2 in advance and accelerates the formation of a dense barrier layer. Compared with the MFA/PU composites, the MCG/PU composites could produce a more stable and dense barrier structure. Water quality tests show that heavy metals do not leak from FA and CG embedded in PU. This work provided a new strategy for the safe and high-value recycling of coal-based solid waste.

Abstract The saltwater hard clam Mercenaria mercenaria (M. mercenaria) as a representative of low-value shellfish, enhancing its flavor quality, is the key to enter the high-end market. Nevertheless, there has not been reported research on the flavor quality of M. mercenaria. This study compared the flavor quality of selective and non-selective saltwater hard clams of M. mercenaria by using various indicators: proximate component, free amino acids, nucleotides, and metabolomic analysis. The results indicated that selective breeding contributed to the significant improvement contents of crude protein, flavor-associated free amino acids (glutamic acid, aspartic acid, proline, etc.), and nucleotides (AMP) (P 1 and P

High strength aluminum alloy sheets with excellent formability are necessary for the successful fabrication of complex-shaped components in order to prevent cracking during the forming process. To address this issue, this study investigated the formability of annealed (O) and water-quenched (WQ) Al–Cu–Li alloys at both room and cryogenic temperatures. Our results reveal a significant increase in the forming limit curve, with a 40% enhancement observed as the forming temperature decreased from 298 K to 113 K. Additionally, a 30% increase in formability was observed when transitioning from the O temper to the WQ temper. Thickness measurements reveal that WQ pre-treatment and cryogenic forming promoted continuous work hardening, thus improving deformation homogeneity. The mechanism responsible for the higher work hardening ability was elucidated by examining the microstructure evolution in different regions of the components. The results show that coarse particles in the O tempered alloys accelerated cross slip, resulting in the formation of recovered microstructures (i.e., dislocation cells and Cube texture) while simultaneously hindering dislocation movement and inducing local stress concentration. Conversely, for the WQ tempered alloy deformed at cryogenic temperature, the dissolution of particles and decreased stacking fault energy at low temperature suppressed cross slip and resulted in well-arranged dislocations within the slip system. Consequently, the combination of WQ pre-treatment and cryogenic forming demonstrates enhanced formability in high strength aluminum alloys, offering a novel method for manufacturing complex-shaped components.

BackgroundThe infiltration and activation of immune cells in the tumor microenvironment (TIME) affect the prognosis of patients with cancer. Tertiary lymphoid structure (TLS) formation favors tumour- infiltrating-lymphocyte (TIL) recruitment and is regarded as an important indicator of good prognosis associated with immunotherapy in patients with tumors. Chemotherapy is currently one of the most commonly used clinical treatment methods. However, there have been no clear report to explore the effects of different types of chemotherapy on TLS formation in the TIME. This study examined the effects of immunogenic cell death (ICD)-inducing chemotherapeutics on immune cells, high-endothelial venules (HEV), and TLSs in mouse melanomas.MethodsDoxorubicin (an ICD inducer), gemcitabine (non-ICD inducer), and a combination of the two drugs was delivered intra-peritoneally to B16F1-loaded C57BL/6 mice. The infiltration of immune cells into tumor tissues was evaluated using flow cytometry. HEV and TLS formation was assessed using immunohistochemistry and multiple fluorescent immunohistochemical staining.ResultsDoxorubicin alone, gemcitabine alone, and the two-drug combination all slowed tumor growth, with the combined treatment demonstrating a more pronounced effect. Compared with the control group, the doxorubicin group showed a higher infiltration of CD8+ T cells and tissue-resident memory T cells (TRM) and an increase in the secretion of interferon-γ, granzyme B, and perforin in CD8+ T subsets and activation of B cells and dendritic cells. Doxorubicin alone and in combination with gemcitabine decreased regulatory T cells in the TIME. Moreover, doxorubicin treatment promoted the formation of HEV and TLS. Doxorubicin treatment also upregulated the expression of programmed cell death protein (PD)-1 in CD8+ T cells and programmed cell death protein ligand (PD-L)1 in tumor cells.ConclusionsThese results indicate that doxorubicin with an ICD reaction promotes TLS formation and increases PD-1/PD-L1 expression in tumor tissues. The results demonstrate the development of a therapeutic avenue using combined immune checkpoint therapy.

This work was devised to discuss the effect of AIM2 on the immunosuppression of LUAD tumors, as well as its molecular mechanism. An allograft mouse model was built. Mouse macrophages were isolated and collected. The infiltration level of Mø and expression of M1 Mø, M2 Mø markers, and PD-L1 were assayed by IHC and flow cytometry. Expression levels of M1 Mø and M2 Mø marker genes and PD-L1 were detected by qPCR. The expression of proteins linked with JAK/STAT3 was tested by western blot. CD8+T cells and NK cells were activated in vitro and co-cultured with mouse macrophages, and their cytotoxicity was detected by LDH method. The proportion of CD206+PD-L1+ cells and the activation and proliferation of CD8+T cells were assayed by flow cytometry. Multicolor immunofluorescence was utilized to assay the co-localization of proteins. AIM2 demonstrated a high expression in LUAD, exhibiting a conspicuous positive correlation with the expression of the M2 Mø markers as well as PD-L1. Expression of M1 markers was upregulated after knockdown of AIM2, while M2 markers expression and PD-L1 were downregulated, and the colocalization of proteins linked with PD-L1 and M2 Mø was decreased. The infiltration and cytotoxicity of CD8+T cells and NK cells increased after silencing AIM2. After the knockdown of AIM2, which was enriched in the JAK/STAT3 pathway, the phosphorylation levels of JAK1, JAK2, and STAT3 were reduced, the immune infiltration level of CD8+T cells increased, and the co-localization level of PD-L1 and PD-1 dropped. The activity and proliferation level of CD8+T cells were increased with the reduced PD-1 expression. AIM2 fosters M2 Mø polarization and PD-L1 expression via the JAK/STAT3 pathway. Moreover, AIM2 promotes the immune escape of LUAD via the PD-1/PD-L1 axis. Our work may blaze a trail for the clinical treatment of LUAD.

Abstract Background Furofuran lignans, the main insecticidal ingredient in Phryma leptostachya, exhibit excellent controlling efficacy against a variety of pests. During the biosynthesis of furofuran lignans, Dirigent proteins (DIRs) are thought to be dominant in the stereoselective coupling of coniferyl alcohol to form ( ±)-pinoresinol. There are DIR family members in almost every vascular plant, but members of DIRs in P. leptostachya are unknown. To identify the PlDIR genes and elucidate their functions in lignan biosynthesis, this study performed transcriptome-wide analysis and characterized the catalytic activity of the PlDIR1 protein. Results Fifteen full-length unique PlDIR genes were identified in P. leptostachya. A phylogenetic analysis of the PlDIRs classified them into four subfamilies (DIR-a, DIR-b/d, DIR-e, and DIR-g), and 12 conserved motifs were found among them. In tissue-specific expression analysis, except for PlDIR7, which displayed the highest transcript abundance in seeds, the other PlDIRs showed preferential expression in roots, leaves, and stems. Furthermore, the treatments with signaling molecules demonstrated that PlDIRs could be significantly induced by methyl jasmonate (MeJA), salicylic acid (SA), and ethylene (ETH), both in the roots and leaves of P. leptostachya. In examining the tertiary structure of the protein and the critical amino acids, it was found that PlDIR1, one of the DIR-a subfamily members, might be involved in the region- and stereo-selectivity of the phenoxy radical. Accordingly, LC–MS/MS analysis demonstrated the catalytic activity of recombinant PlDIR1 protein from Escherichia coli to direct coniferyl alcohol coupling into ( +)-pinoresinol. The active sites and hydrogen bonds of the interaction between PlDIR1 and bis-quinone methide (bisQM), the intermediate in ( +)-pinoresinol formation, were analyzed by molecular docking. As a result, 18 active sites and 4 hydrogen bonds (Asp-42, Ala-113, Leu-138, Arg-143) were discovered in the PlDIR1-bisQM complex. Moreover, correlation analysis indicated that the expression profile of PlDIR1 was closely connected with lignan accumulations after SA treatment. Conclusions The results of this study will provide useful clues for uncovering P. leptostachya's lignan biosynthesis pathway as well as facilitate further studies on the DIR family.

Abstract The combination of PD-1 blockade with neoadjuvant chemotherapy (NAC) has achieved unprecedented clinical success in non-small cell lung cancer (NSCLC) compared to NAC alone, but the underlying mechanisms by which PD-1 blockade augments the effects of chemotherapy remain incompletely elucidated. Single-cell RNA sequencing was performed on CD45+ immune cells isolated from surgically resected fresh tumors of seven NSCLC patients receiving NAC or neoadjuvant pembrolizumab and chemotherapy (NAPC). Multiplex fluorescent immunohistochemistry was performed on FFPE tissues before and after NAC or NAPC from 65 resectable NSCLC patients, and results were validated with GEO dataset. NAC resulted in an increase only of CD20+ B cells, whereas NAPC increased the infiltration of CD20+ B cells, CD4+ T cells, CD4+CD127+ T cells, CD8+ T cells, CD8+CD127+ and CD8+KLRG1+ T cells. Synergistic increase in B and T cells promotes favorable therapeutic response after NAPC. Spatial distribution analysis discovered that CD8+ T cells and their CD127+ and KLRG1+ subsets were in closer proximity to CD4+ T/CD20+ B cells in NAPC versus NAC. GEO dataset validated that B-cell, CD4, memory, and effector CD8 signatures correlated with therapeutic responses and clinical outcomes. The addition of PD-1 blockade to NAC promoted anti-tumor immunity through T and B cells recruitment in the tumor microenvironment and induced tumor-infiltrating CD8+ T cells skewed toward CD127+ and KLRG1+ phenotypes, which may be assisted by CD4+ T cells and B cells. Our comprehensive study identified key immune cell subsets exerting anti-tumor responses during PD-1 blockade therapy and that may be therapeutically targeted to improve upon existing immunotherapies for NSCLC.

Starch digestion rate and location in the gastrointestinal tract (GIT) are critical for human health. This review aims to present a comprehensive summary on our current understanding of physiological, biochemical, anatomical and geometrical factors of human digestive system that are related to in vivo starch digestibility. It is shown that all digestive compartments including mouth, stomach, small intestine, and large intestine play critical roles in regulating the overall starch digestion process. A proper investigation of starch digestion pattern should thus be based on the consideration of all these compartments. Main biological factors are summarized as oral mastication and salivation, gastric emptying and motility, small intestinal enzymes and motility, large intestinal resistant starch (RS)-microbiota interactions and gut-brain feedback control, as well as glucose adsorption and hormonal feedback control. However, connections among these biological factors in determining starch digestive behaviors remain elusive. This is due to the inherent complexity of human GIT anatomy, motility and biochemical conditions, as well as ethical, financial and technical issues in conducting clinical studies. Much technological and scientific efforts from both clinical studies and in vitro simulation models are required for a better understanding of in vivo starch digestion behaviors.

Waste tire rubber powder-modified asphalt (RMA) has been widely used in road construction, which was traditionally limited by the poor compatibility of RMA, affecting pavement performance. By synthesizing epoxy soybean oil with amide groups (ESO/TETA) and grafting it onto desulfurized rubber powder (DCR) through microwave irradiation, a surface-functionalized rubber powder (MDCR) was produced successfully. The effects of the physical properties, storage stability, thermal stability, and rheological behavior of the modified asphalt were studied. The results show that the MDCR with a polar surface improved the compatibility and adhesive interactions between the modified crumb rubber and the asphalt. The MDCR content could reach 50%, and the phase separation could meet the requirements of 2.2 °C, which has application conditions in engineering for stable storage. Additionally, the inclusion of MDCR in the asphalt formulations significantly mitigated the temperature sensitivity of the modified asphalt. Importantly, when the MDCR constituted from 20% to 50% of the asphalt, there was a noted reduction in the phase angle at temperatures above 70 °C, indicating a significant improvement in the elastic efficiency. The MDCR also led to substantial enhancements in the resistance of the asphalt to high-temperature and high-stress rutting, addressing the crucial limitations in the consumption ability of waste tire rubber powder and improving the overall performance of RMA in pavement applications.

Background: Oncogenic PIK3CA mutations (PIK3CAmut) frequently occur in a higher proportion in luminal breast cancer (LBC), especially in refractory advanced cases, and are associated with changes in tumour cellular metabolism. Nevertheless, its effect on the progression of the immune microenvironment (TIME) within tumours and vital molecular events remains veiled. Methods: Multiplex immunohistochemistry (mIHC) and single‐cell mass cytometry (CyTOF) was used to describe the landscape of TIME in PIK3CAmut LBC. The PIK3CA mutant cell lines were established using CRISPER/Cas9 system. The gene expression levels, protein secretion and activity of signaling pathways were measured by real‐time RT‐PCR, ELISA, immunofluorescence staining or western blotting. GSEA analysis, transwell chemotaxis assay, live cell imaging, flow cytometry metabolite analysis targeting arachidonic acid, Dual‐luciferase reporter assay, and Chromatin immunoprecipitation assay were used to investigate the underlying function and mechanism of the PI3K/5‐LOX/LTB4 axis. Results: PIK3CAmut LBC cells can induce an immunosuppressive TIME by recruiting myeloid‐derived suppressor cells (MDSCs) and excluding cytotoxic T cells via the arachidonic acid (AA) metabolism pathway. Mechanistically, PIK3CAmut activates the transcription of 5‐lipoxygenase (5‐LOX) in a STAT3‐dependent manner, which in turn directly results in high LTB4 production, binding to BLT2 on MDSCs and promoting their infiltration. Since a suppressive TIME is a critical barrier for the success of cancer immunotherapy, the strategies that can convert “cold” tumours into “hot” tumours were compared. Targeted therapy against the PI3K/5‐LOX/LTB4 axis synergizing with immune checkpoint blockade (ICB) therapy achieved dramatic shrinkage in vivo. Conclusions: The results emphasize that PIK3CAmut can induce immune evasion by recruiting MDSCs through the 5‐LOX‐dependent AA pathway, and combination targeted therapy with ICB may provide a promising treatment option for refractory advanced LBC patients.

The Ag@MgO nanoparticles (NPs), consisting of silver NPs with a size range of 6–15 nm well-dispersed on the surface of MgO nanoparticles, were synthesized using a simple and efficient in situ synthesis method. These NPs exhibited superior antibacterial activity compared to bare MgO NPs. Subsequently, they were incorporated into poly(butylene succinate-co-terephthalate) (PBST) matrix to prepare antibacterial food packaging films. The effects of Ag@MgO NPs morphology on the compatibility, mechanical, water vapor barrier, UV blocking and antibacterial properties of PBST matrix were evaluated. Unmodified Ag@MgO NPs caused agglomeration and poor mechanical properties. However, when the Ag@MgO NPs were surface modified and interacted with the PBST matrix, the dispersion of nanoparticles and crystallinity improved, resulting in enhanced tensile strength and barrier properties of the composite films. Furthermore, the well-formed of Ag@MgO NPs in the films played a positive role in controlling the growth of microorganisms. Upon incorporation of the modified Ag@MgO NPs, the PBST nanocomposite films exhibited a 23.5% increase in the tensile strength and a 63% decrease in the water vapor permeability compared to PBST films. Moreover, the UV blocking performances were enhanced, and high antibacterial rates of 99.8% and 99.7% were achieved against E. coli and S. aureus, respectively. All the results indicate that the PBST incorporating modified Ag@MgO NPs have excellent potential for use in food packaging applications.

Objectives: Immune checkpoint (ICP) expression in tumor cells could directly or indirectly affect the results of immunotherapy. ICP ligands on tumor cells usually bind their immune cell receptors to inhibit the activity, resulting in tumor immune escape. Thus, the purpose of this study was to ascertain the impact of various chemotherapeutic drugs on ICP expression in non-small cell lung cancer (NSCLC) cell lines with different pathological subtypes to provide a basis for the development of a superior regimen of chemotherapy combined with ICP blockade. Methods: Several first-line chemotherapy agents (cisplatin, carboplatin, paclitaxel, gemcitabine, vinorelbine, and pemetrexed) were selected to treat different NSCLC cell lines (squamous carcinoma H1703, adenocarcinoma A549, and large cell cancer H460) for 72 hours, and then the changes in ICP expression in the tumor cells were observed through flow cytometry. Results: Cisplatin, carboplatin, and paclitaxel upregulated the expressions of programmed cell death ligand 1 (PD-L1) and programmed cell death ligand 2 (PD-L2) in A549 and H460 cell lines. Meanwhile, vinorelbine and pemetrexed upregulated PD-L1 and PD-L2 in H1703, A549, and H460 cell lines. Paclitaxel, gemcitabine, vinorelbine, and pemetrexed significantly upregulated the expressions of both galectin-9 and high-mobility group box protein 1 (HMGB1) in the A549 cell line. Cisplatin and paclitaxel significantly upregulated the expressions of major histocompatibility complex-II (MHC-II), galectin-3, α-synuclein, and fibrinogen-like protein 1 (FGL1) in A549 and H460 cell lines. In addition, cisplatin and vinorelbine significantly upregulated the expressions of both CD155 and CD112 in the H460 cell line. Vinorelbine upregulated MHC-I in all three cell lines. Conclusion: Chemotherapy agents have different effects on the expression of ICP ligands in tumor cells with different pathological types, and this may affect the efficacy of combined immunotherapy. These results provide a theoretical basis for further selection and optimization of the combination of chemotherapy and immunotherapy.

Background and objective Small cell lung cancer (SCLC) with high c-Myc expression is prone to relapse and metastasis, leading to extremely low survival rate. Cyclin-dependent kinases 4 and 6 (CDK4/6) inhibitor Abemaciclib plays a key role in the treatment of tumors, but the effects and mechanisms on SCLC remain unclear. This study was to analyze the effect and molecular mechanism of Abemaciclib in inhibiting proliferation, migration and invasion of SCLC with high c-Myc expression, with a view to expanding a new direction for reducing the recurrence and metastasis. Methods Proteins interacting with CDK4/6 were predicted using the STRING database. The expressions of CDK4/6 and c-Myc in 31 cases of SCLC cancer tissues and paired adjacent normal tissues were analyzed by immunohistochemistry. The effects of Abemaciclib on the proliferation, invasion and migration of SCLC were detected by CCK-8, colony formation assay, Transwell and migration assay. Western blot was used to detect the expressions of CDK4/6 and related transcription factors. Flow cytometry was used to analyze the effects of Abemaciclib on the cell cycle and checkpoint of SCLC. Results The expression of CDK4/6 was associated with c-Myc by STRING protein interaction network. c-Myc can directly modalize achaete-scute complex homolog 1 (ASCL1), neuronal differentiation 1 (NEUROD1) and Yes-associated protein 1 (YAP1). Moreover, CDK4 and c-Myc regulate the expression of programmed cell death ligand 1 (PD-L1). Immunohistochemistry showed that the expressions of CDK4/6 and c-Myc in cancer tissues were higher than those in adjacent tissues(P

Abstract Background Subclinical anxiety, depressive and somatic symptoms appear closely related. However, it remains unclear whether somatic symptoms mediate the association between subclinical anxiety and depressive symptoms and what the underlying neuroimaging mechanisms are for the mediating effect. Methods Data of healthy participants (n = 466) and participants in remission of major depressive disorder (n = 53) were obtained from the Human Connectome Project. The Achenbach Adult Self-Report was adopted to assess anxiety, depressive and somatic symptoms. All participants completed four runs of resting-state functional magnetic resonance imaging. Mediation analyses were utilized to explore the interactions among these symptoms and their neuroimaging mechanisms. Results Somatic symptoms partially mediated the association between subclinical anxiety and depressive symptoms in healthy participants (anxiety→somatic→depression: effect: 0.2785, Boot 95% CI: 0.0958–0.3729; depression→somatic→anxiety: effect: 0.0753, Boot 95% CI: 0.0232–0.1314) and participants in remission of MDD (anxiety→somatic→depression: effect: 0.2948, Boot 95% CI: 0.0357–0.7382; depression→somatic→anxiety: effect: 0.0984, Boot 95% CI: 0.0007–0.2438). Resting-state functional connectivity (FC) between the right medial superior frontal gyrus and the left thalamus and somatic symptoms as chain mediators partially mediated the effect of subclinical depressive symptoms on subclinical anxiety symptoms in healthy participants (effect: 0.0020, Boot 95% CI: 0.0003–0.0043). The mean strength of common FCs of subclinical depressive and somatic symptoms, somatic symptoms, and the mean strength of common FCs of subclinical anxiety and somatic symptoms as chain mediators partially mediated the effect of subclinical depressive symptoms on subclinical anxiety symptoms in remission of MDD (effect: 0.0437, Boot 95% CI: 0.0024–0.1190). These common FCs mainly involved the insula, precentral gyri, postcentral gyri and cingulate gyri. Furthermore, FC between the triangular part of the left inferior frontal gyrus and the left postcentral gyrus was positively associated with subclinical anxiety, depressive and somatic symptoms in remission of MDD (FDR-corrected p

The aim of this study was to investigate the improvement effect of mussel polysaccharide α-D-glucan (MP-A) on non-alcoholic fatty liver disease (NAFLD) in the high-fat diet (HFD) induced apolipoprotein E knockout (ApoE−/−) mice. 20 normal C57BL/6J mice were randomly divided into two groups: Normal group, normal+MP-A group, and 20 ApoE−/− mice were randomly divided into two groups: Model group, model+MP-A group, 10 mice in each group. ApoE−/− mice were fed with high fat diet to induce the NAFLD model. After MP-A were administered, serum and liver triglyceride (TG) and total cholesterol (TC) levels were detected, and the pathological morphology of liver tissue were evaluated by histopathology. The mRNA levels and protein levels of the receptors related to lipid metabolism and cholesterol metabolism were also measured simultaneously. Results showed that compared with the model group, the mice in the model+MP-A group showed significant decreases in the weight growth rate (P

The graphene-modified composite poly(lactic acid) (PLA)/poly(butylene adipate-co-terephthalate) (PBAT) monofilaments were prepared by melt spinning. The effects of graphene (GR) contents on the microstructural, thermal, dynamic mechanical, and mechanical properties and the degradation behavior of the graphene-modified composite PLA/PBAT monofilaments were studied. Scanning electron microscopy (SEM) confirmed the improved compatibility of the PLA particles in the PBAT matrix, while the transmission electron microscopy (TEM) demonstrated the size and morphology of the GR distribution in the polymer. As the GR content increased, dynamic mechanical analysis (DMA) clarified a decrease of the difference of glass transition temperature (Tg) between PLA and PBAT. In addition, the mechanical properties of the composite monofilaments were significantly enhanced at a GR content of 0.2 wt%. Weight loss rate of the composite monofilaments under soil and sea degradation conditions were obviously related to the GR content.

BackgroundLower respiratory tract infections are the leading cause of morbidity and mortality in children worldwide. Timely and accurate pathogen detection is crucial for proper clinical diagnosis and therapeutic strategies. The low detection efficiency of conventional methods and low specificity using respiratory samples seriously hindered the accurate detection of pathogens.MethodsIn this study, we retrospectively enrolled 1,032 children to evaluate the performance of metagenomics next-generation sequencing (mNGS) using bronchoalveolar lavage fluid (BALF) sample and protected bronchial brushing (BB) sample in diagnosing pneumonia in children. In addition, conventional tests (CTs) were also performed.ResultsThe specificity of BB mNGS [67.3% (95% CI 58.6%–75.9%)] was significantly higher than that of BALF mNGS [38.5% (95% CI 12.0%–64.9%)]. The total coincidence rate of BB mNGS [77.6% (95% CI 74.8%–80.5%)] was slightly higher than that of BALF mNGS [76.5% (95% CI 68.8%–84.1%)] and CTs [38.5% (95% CI 35.2%–41.9%)]. During the epidemics of Mycoplasma pneumoniae, the detection rate of M. pneumoniae in the >6-year group (81.8%) was higher than that in the 3–6-year (78.9%) and 6-year, and 3–6-year groups, respectively. mNGS detection should be performed at the duration of 5–7 days after the start of continuous anti-microbial therapy or at the duration of 6–9 days from onset to mNGS test.ConclusionsThis is the first report to evaluate the performance of BB mNGS in diagnosing pulmonary infections in children on a large scale. Based on our findings, extensive application of BB mNGS could be expected.

Chinese steamed bread (CSB), which is widely consumed in East Asia, usually undergoes storage before consumption, but it is unclear how different storage temperatures affect CSB starch retrogradation and digestion properties, which are important for consumers. CSB was stored for 2 days at 25 °C, 4 °C, −18 °C, 4 °C/25 °C temperature cycling (i.e., 24 h at 4 °C, followed by 24 h at 25 °C) and −18 °C/ 25 °C temperature cycling. The results revealed for the first time that more orderly starch double helices are formed when CSB was stored at 4 °C or 4 °C/25 °C. Storage under −18 °C produced lower amounts of, but more heterogenous, starch double helices, with fewer B-type, but more V-type, crystallites. Compared to other storage temperatures, more long-range intermolecular interactions formed between the starch and protein at 4 °C or 4 °C/25 °C. CSB samples showed the slowest starch digestibility when stored at 4 °C. The impact of storage temperature on the starch retrogradation properties and digestibility of CSB also depended on the wheat variety, attributed to differences in the starch molecular structure. These results have significance and practical applications to help the CSB food industry to control starch retrogradation and digestibility. For example, CSB could be stored at 4 °C for 2 days in order to reduce its starch digestibility.

Abstract The combination of immune checkpoint inhibitors (ICIs) with chemotherapy (chemoimmunotherapy) in the neoadjuvant setting have achieved favorable clinical benefits in non-small cell lung cancer (NSCLC), but the mechanism of clinical responses remain unclear. We provide a rich resource of 186,477 individual immune cells from 48 samples of four treatment-naive and eight neoadjuvant chemoimmunotherapy treated IIIA NSCLC patients (responders versus non-responders) by single-cell RNA-seq and TCR-seq. We observed the synergistic increase of B cells and CD4+ T cells were associated with a positive therapeutic response of neoadjuvant chemoimmunotherapy. B cell IgG subclasses IgG1 and IgG3 played a critical role in anti-tumor immune response in tumor lesions, and this process was driven by increased IL-21 secreted by infiltrated T follicular helper (Tfh) cells after neoadjuvant chemoimmunotherapy. Furthermore, we uncovered several critical events for positive clinical outcomes, including the diminished activated TNFRSF4+ regulatory T cells (Tregs), increased LAMP3+ dendritic cells (DCs), and the expansion of intratumoral CD4+ T clones and peripheral C3-Cytotoxic CD8+ T clones. A validation cohort of 26 treatment-naive and 30 neoadjuvant chemoimmunotherapy treated IIIA/ IIIB NSCLC patients verified these findings. In total, our comprehensive study of the single-cell profile of immune cells provides insights into mechanisms underlying anti-PD-1-based therapies and identified potential predictive factors and therapeutic targets for improving the efficiency of neoadjuvant chemoimmunotherapy in NSCLC.

Background and goalNoninvasive prediction of isocitrate dehydrogenase (IDH) mutation status in glioma guides surgical strategies and individualized management. We explored the capability on preoperatively identifying IDH status of combining a convolutional neural network (CNN) and a novel imaging modality, ultra-high field 7.0 Tesla (T) chemical exchange saturation transfer (CEST) imaging.MethodWe enrolled 84 glioma patients of different tumor grades in this retrospective study. Amide proton transfer CEST and structural Magnetic Resonance (MR) imaging at 7T were performed preoperatively, and the tumor regions are manually segmented, leading to the “annotation” maps that offers the location and shape information of the tumors. The tumor region slices in CEST and T1 images were further cropped out as samples and combined with the annotation maps, which were inputted to a 2D CNN model for generating IDH predictions. Further comparison analysis to radiomics-based prediction methods was performed to demonstrate the crucial role of CNN for predicting IDH based on CEST and T1 images.ResultsA fivefold cross-validation was performed on the 84 patients and 4090 slices. We observed a model based on only CEST achieved accuracy of 74.01% ± 1.15%, and the area under the curve (AUC) of 0.8022 ± 0.0147. When using T1 image only, the prediction performances dropped to accuracy of 72.52% ± 1.12% and AUC of 0.7904 ± 0.0214, which indicates no superiority of CEST over T1. However, when we combined CEST and T1 together with the annotation maps, the performances of the CNN model were further boosted to accuracy of 82.94% ± 1.23% and AUC of 0.8868 ± 0.0055, suggesting the importance of a joint analysis of CEST and T1. Finally, using the same inputs, the CNN-based predictions achieved significantly improved performances above those from radiomics-based predictions (logistic regression and support vector machine) by 10% to 20% in all metrics.Conclusion7T CEST and structural MRI jointly offer improved sensitivity and specificity of preoperative non-invasive imaging for the diagnosis of IDH mutation status. As the first study of CNN model on imaging acquired at ultra-high field MR, our results could demonstrate the potential of combining ultra-high-field CEST and CNN for facilitating decision-making in clinical practice. However, due to the limited cases and B1 inhomogeneities, the accuracy of this model will be improved in our further study.

Anastomotic leakage is a serious complication following gastrointestinal surgery and one of the leading causes of patient mortality. Despite the significant clinical and economic burden, there are currently no reliable treatment options to improve the healing of intestinal anastomosis and subsequently prevent anastomotic leakage. Recently, the development of regenerative medicine has shown promise for improving anastomotic healing. Recent studies have illustrated that stem cell-derived secretome can enhance tissue regeneration without the safety and ethical limitations of stem cell transplantation. Herein, we developed a fibrin glue topical delivery system loaded with mesenchymal stem cells (MSCs)-derived secretome for controlled delivery of bioactive factors, and evaluated its application potential in improving the healing of intestinal anastomosis. Under in vitro conditions, the MSCs secretome significantly promoted cell proliferation viability in a dose-dependent manner and resulted in the controlled release of growth factors via fibrin glue delivery. We established a rat surgical anastomotic model and experimentally found that MSCs secretome-loaded fibrin glue enhanced anastomotic bursting pressure, increased granulation tissue formation and collagen deposition, and significantly promoted anastomotic healing. Mechanistically, fibrin glue accelerated cell proliferation, angiogenesis, and macrophage M2 polarization at the surgical anastomotic site by releasing bioactive factors in the secretome, and it also alleviated the inflammatory response and cell apoptosis at the anastomotic site. Our results demonstrated for the first time that MSCs-derived secretome could promote the healing of intestinal anastomosis. Considering the accessibility and safety of the cell-free secretome, we believed that secretome-loaded fibrin glue would be a cell-free therapy to accelerate the healing of intestinal anastomosis with great potential for clinical translation.

Herein, Liu and colleagues show that CD4+ T cells in cytokine-induced killer (CIK) cells are required to enhance the clinical efficacy of CIK therapy. In addition, CD4+ T cells help via IL-17A production is critical to restoring the function of CD8+/CD3+CD56+ T cells and reversing the exhaustion of PD-1+TIM-3+ T cells in CIK therapy.

Effect of inorganic fillers, cement powder, Na-bentonite, flay ash, on curing kinetics of bisphenol-A epoxy resin and polyamide (BAE/PA) system was investigated under iso-thermal curing condition using the differential scanning calorimeter (DSC). Meanwhile, their thixotropic properties were measured using a rheometer at 25 °C. The relation between curing kinetics and thixotropic properties was discussed. It is found that when BAE/PA system is cured under 50–100 °C, the overall reaction order (m + n) decreases with increasing temperature from 2.63 to 1.83. The whole curing reaction process can be described by the Kamal model at temperatures above 90 °C. When cured below 80 °C, the curing process changed from chemical-controlled reaction into diffusion-controlled reaction at the critical curing degree (αc). αc decreases with the decrease of curing temperature. When filled by different kinds of fillers, curing kinetics at 90 °C and thixotropic properties of the systems at 25 °C were studied. It is found that compared with fly ash and cement powder, the system with Na-bentonite has the largest thixotropic hysteresis hoop area and changes into diffusion control reaction at lower curing degree. Decreasing particle size and increasing mass fraction of Na-bentonite increased the thixotropic properties of the filled system and consequently improved the overall reaction order. It means that high surface activity and thickening effect of Na-bentonite make the epoxy-amine system shows a stronger autocatalytic effect.

Abstract Background Early clinical trials indicate that patients with anaplastic lymphoma kinase (ALK)‐driven non‐small cell lung cancer (NSCLC) have a lower response rate to programmed cell death protein 1 (PD‐1) antibody therapy. However, the specific mechanism underlying this remains unclear. To further explore the characteristics of the tumor microenvironment and determine the potential mechanism of immunotherapy resistance in patients with ALK, we selected another important immune‐related molecule, major histocompatibility complex class I (HLA‐I), as the focus of our study. Methods We collected the biopsy samples of 140 patients with NSCLC. The number of CD8+ T cells and HLA‐I/programmed cell death 1 ligand 1 (PD‐L1) expression were determined by immunohistochemistry. Disease‐free survival (DFS) and overall survival (OS) were analyzed using the Kaplan–Meier method, and their relationship with patient clinical characteristics analyzed using Cox proportional hazards regression. In addition, we treated ALK‐positive lung cancer cells with ALK inhibitors in vitro to observe changes of HLA‐I. Results ALK positivity was associated with low membrane PD‐L1 and HLA‐I expression. However, these two indicators were not associated with the prognosis of patients with stage I–IIIa NSCLC. Inhibition of ALK could upregulate HLA‐I membrane expression to a certain extent. Conclusion Patients with ALK fusion showed downregulation of PD‐L1 and HLA‐I expression on the tumor cell membrane. Inhibition of ALK and its downstream signaling pathway can reverse it. These results suggest that the appropriate combination therapy should be considered for patients with ALK fusion and using targeted therapy at the proper time may increase patient benefits.

Objective: Trained immunity of natural killer (NK) cells has shown great potential in the treatment of cancers by eliciting enhanced effector responses to restimulation by cytokines or cancer cells for long time periods after preactivation. However, the human NK cells responsible for the generation and maintenance of trained immunity are largely unknown. We hypothesized that heterogeneous human NK cells would respond differentially to stimulation with a combination of IL-12, IL-15, and IL-18, and that an NK cell subset might exist that is mainly responsible for the induction of trained immunity. On the basis of our hypothesis, we aimed to identify the subset from which cytokine-trained human NK cells originate and to explore possible regulatory targets for drug intervention. Methods: Flow cytometry assays were performed to analyze the functions of cytokine-trained NK cells and examine cell division and protein expression in NK cell subsets. Single-cell RNA sequencing (scRNA-seq) plus TotalSeq™ technology was used to track the heterogeneity of NK cells during the induction of trained immunity. Results: Traditional developmental markers for peripheral NK cells were unable to identify the precursors of human NK cells with trained immunity. Therefore, we used scRNA-seq plus TotalSeq™ technology to track the heterogeneity of NK cells during the induction of trained immunity and identified a unique cluster of CD57−NKG2A+EZH2+IFNG+MKI67+IL12R+IL15R+IL18R+ NK cells. Enrichment and pseudotime trajectory analyses suggested that this cluster of NK cells contained the precursor of trained NK cells. We then used flow cytometry to further investigate the role of EZH2 in trained NK precursors and found that CD57−NKG2A+EZH2+ NK cells had faster cell cycles and an enhanced trained phenotype, and EZH2 inhibition significantly impaired the induction of trained immunity in NK cells. These results suggested that EZH2 is a unique epigenetic marker of precursors of human NK cells with trained immunity. Conclusions: Our work revealed human NK heterogeneity in the induction of trained immunity, identified the precursor subset for trained NK cells, and demonstrated the critical role of EZH2 in the induction of trained immunity in human NK cells.

Objective: Patients with non–small cell lung cancer (NSCLC) respond differently to cytokine-induced killer cell (CIK) treatment. Therefore, potential prognostic markers to identify patients who would benefit from CIK treatment must be elucidated. The current research aimed at identifying predictive prognostic markers for efficient CIK treatment of patients with NSCLC. Methods: Patients histologically diagnosed with NSCLC were enrolled from the Tianjin Medical University Cancer Institute and Hospital. We performed whole-exome sequencing (WES) on the tumor tissues and paired adjacent benign tissues collected from 50 patients with NSCLC, and RNA-seq on tumor tissues of 17 patients with NSCLC before CIK immunotherapy treatment. Multivariate Cox proportional hazard regression analysis was used to analyze the association between clinical parameters and prognostic relevance. WES and RNA-seq data between lung squamous cell carcinoma (SCC) and adenocarcinoma (Aden) were analyzed and compared. Results: The pathology subtype of lung cancer was the most significantly relevant clinical parameter associated with DFS, as analyzed by multivariate Cox proportional hazard regression (P = 0.031). The patients with lung SCC showed better CIK treatment efficacy and extended DFS after CIK treatment. Relatively low expression of HLA class II genes and checkpoint molecules, and less immunosuppressive immune cell infiltration were identified in the patients with lung SCC. Conclusions: Coordinated suppression of the expression of HLA class II genes and checkpoint molecules, as well as less immune suppressive cell infiltration together contributed to the better CIK treatment efficacy in lung SCC than lung Aden.

The present study examined deep gray matter perfusion in neonates with congenital heart disease (CHD) with arterial spin labeling magnetic resonance imaging preoperatively. We found that neonates with cyanotic CHD showed lower right thalamus compared with controls and lower right basal ganglia perfusion compared with acyanotic CHD. When the CHD group was assessed as a whole, it showed slightly decreased left thalamus perfusion compared with controls. The results suggest that cardiac physiology plays a crucial part in changes in regional cerebral perfusion. Perfusion with arterial spin labeling may be a useful marker of high risk for impaired cerebral blood flow auto-regulation and cerebral hypoperfusion in neonates with CHD.

To improve heat dissipation capability and enhance mechanical properties, a series of silica aerogel (SA) and modified glass fiber (GF)-filled SBR composites were prepared. It was found that the addition of SA successfully reduced the thermal conductivity of SBR by 35%, owing to the heat shield of the nanoscale porous structure of SA. Moreover, the addition of modified glass fiber (MGF) yielded a significant increase in the tensile and tear strength of SBR/SA composite rubber of 37% and 15%, respectively. This enhancement was more pronounced than the improvement observed with unmodified GF, and was attributed to the improved dispersion of fillers and crosslinking density of the SBR matrix. Rheological analysis revealed that the addition of SA and MGF weakened the ω dependence. This was due to the partial relaxation of immobilized rubber chains and limited relaxation of rubber chains adsorbed on the MGF. Furthermore, the strain amplification effect of MGF was stronger than that of GF, leading to a more pronounced reinforcing effect.

The accurate and effective detection of SF6 decomposition components inside a gas-insulated switchgear (GIS) is crucial for equipment fault diagnosis and condition assessment. The current method for detecting SF6 decomposition components involves gas extraction at the GIS inlet, which only provides limited information on the decomposition component content. Therefore, there is a need to explore more effective ways to obtain internal gas component information within GIS. In this study, we propose a graphene-coated microfiber gas detection method for SO2. We establish a physical simulation model of the microfiber and analyze the sensing mechanism of the microfiber diameter and cladding refractive index changes in its evanescent field. A graphene-coated microfiber gas sensor was prepared using a drop-coating method, and a fiber loop ring-down (FLRD) gas detection system was constructed for the experimental studies on SO2 gas detection. The results demonstrated that the graphene-coated microfiber exhibits an excellent gas-sensitive response to SO2 and achieves trace-level detection at room temperature. The concentration range of 0 to 200 ppm showed good linearity, with a maximum detection error of 4.76% and a sensitivity of 1.24 ns/ppm for SO2. This study introduces an all-fiber method for detecting SF6 decomposition components, offering a new approach for online monitoring of SF6 decomposition components in GIS equipment using built-in fiber-optic sensors.

Cephalometric analysis is a standard diagnostic tool in orthodontics and craniofacial surgery. Today, as conventional 2D cephalometry is limited and susceptible to analysis bias, a more reliable and user-friendly three-dimensional system that includes hard tissue, soft tissue, and airways is demanded in clinical practice. We launched our study to develop such a system based on CT data and landmarks. This study aims to determine whether the data labeled through our process is highly qualified and whether the soft tissue and airway data derived from CT scans are reliable. We enrolled 15 patients (seven males, eight females, 26.47 ± 3.44 years old) diagnosed with either non-syndromic dento–maxillofacial deformities or OSDB in this study to evaluate the intra- and inter-examiner reliability of our system. A total of 126 landmarks were adopted and divided into five sets by region: 28 cranial points, 25 mandibular points, 20 teeth points, 48 soft tissue points, and 6 airway points. All the landmarks were labeled by two experienced clinical practitioners, either of whom had labeled all the data twice at least one month apart. Furthermore, 78 parameters of three sets were calculated in this study: 42 skeletal parameters (23 angular and 19 linear), 27 soft tissue parameters (9 angular and 18 linear), and 9 upper airway parameters (2 linear, 4 areal, and 3 voluminal). Intraclass correlation coefficient (ICC) was used to evaluate the inter-examiner and intra-examiner reliability of landmark coordinate values and measurement parameters. The overwhelming majority of the landmarks showed excellent intra- and inter-examiner reliability. For skeletal parameters, angular parameters indicated better reliability, while linear parameters performed better for soft tissue parameters. The intra- and inter-examiner ICCs of airway parameters referred to excellent reliability. In summary, the data labeled through our process are qualified, and the soft tissue and airway data derived from CT scans are reliable. Landmarks that are not commonly used in clinical practice may require additional attention while labeling as they are prone to poor reliability. Measurement parameters with values close to 0 tend to have low reliability. We believe this three-dimensional cephalometric system would reach clinical application.