P2RX1-Negative neutrophils promote the immunosuppressive microenvironment in Non-Small cell lung cancer by Upregulating PD-L1 expression.

BACKGROUND: The most abundant innate immune cells, neutrophils, contribute significantly to cancer development by stimulating immunosuppression. However, it remains unclear about its function and molecular mechanisms in the immunosuppressive microenvironment of non-small cell lung cancer (NSCLC). METHODS: Blood samples were collected from NSCLC patients and healthy volunteers to detect the expression of P2RX1 and PD-L1 in neutrophils using qRT-PCR, western blot (WB), and flow cytometry. Neutrophils were sorted into P2RX1-positive (P2RX1+)/P2RX1-negative (P2RX1-) groups and co-cultured with CD8+ T cells. Changes in the proliferative and cytotoxic capabilities of CD8+ T cells were then detected using flow cytometry and enzyme-linked immunosorbent assay. The content of granzyme B was determined by enzyme-linked immunosorbent assay. The effects of P2RX1-deficient neutrophils on fatty acids, triglycerides, lipid droplet content and FASN expression were detected using kits, Nile red staining and WB, respectively. RESULTS: This study revealed a deficiency in P2RX1 expression in peripheral blood neutrophils of NSCLC patients, which was negatively correlated with the expression of PD-L1. P2RX1-neutrophils inhibited T cell proliferation and granzyme B expression and promoted T cell exhaustion. Furthermore, in P2RX1-deficient neutrophils, there was a notable increase in the levels of fatty acids, triglycerides, and lipid droplet accumulation, as well as an upregulation of FASN protein expression. Mechanistically, P2RX1-neutrophils upregulated PD-L1 expression by inducing fatty acid metabolism to improve immunosuppression in NSCLC. CONCLUSION: The mechanism by which P2RX1-deficient neutrophils contributed to immunosuppressive effects in NSCLC was clarified by our findings, indicating that P2RX1 could be one potential target for counteracting the immunosuppressive effects of neutrophils.

Phenotyping of FGF12AV52H mutation in mouse implies a complex FGF12 network.

Pathogenic missense mutation of the FGF12 gene is responsible for a variable disease phenotypic spectrum. Disease-specific therapies require precise dissection of the relationship between different mutations and phenotypes. The lack of a proper animal model hinders the investigation of related diseases, such as early-onset epileptic encephalopathy. Here, an FGF12AV52H mouse model was generated using CRISPR/Cas9 technology, which altered the A isoform without affecting the B isoform. The FGF12AV52H mice exhibited seizure susceptibility, while no spontaneous seizures were observed. The increased excitability in dorsal hippocampal CA3 neurons was confirmed by patch-clamp recordings. Furthermore, immunostaining showed that the balance of excitatory/inhibitory neurons in the hippocampus of the FGF12AV52H mice was perturbed. The increases in inhibitory SOM+ neurons and excitatory CaMKII+ neurons were heterogeneous. Moreover, the locomotion, anxiety levels, risk assessment behavior, social behavior, and cognition of the FGF12AV52H mice were investigated by elevated plus maze, open field, three-chamber sociability, and novel object tests, respectively. Cognition deficit, impaired risk assessment, and social behavior with normal social indexes were observed, implying complex consequences of V52H FGF12A in mice. Together, these data suggest that the function of FGF12A in neurons can be immediate or long-term and involves modulation of ion channels and the differentiation and maturation of neurons. The FGF12AV52H mouse model increases the understanding of the function of FGF12A, and it is of great importance for revealing the complex network of the FGF12 gene in physiological and pathological processes.

Conjugated Linoleic Acid Production in Pine Nut Oil: A Lactiplantibacillus plantarum Lp-01 Fermentation Approach.

Conjugated linoleic acid (CLA) is a class of bioactive fatty acids that exhibit various physiological activities such as anti-cancer, anti-atherosclerosis, and lipid-lowering. It is an essential fatty acid that cannot be synthesized by the human body and must be derived from dietary sources. The natural sources of CLA are limited, predominantly relying on chemical and enzymatic syntheses methods. Microbial biosynthesis represents an environmentally benign approach for CLA production. Pine nut oil, containing 40-60% linoleic acid, serves as a promising substrate for CLA enrichment. In the present study, we developed a novel method for the production of CLA from pine nut oil using Lactiplantibacillus plantarum (L. plantarum) Lp-01, which harbors a linoleic acid isomerase. The optimal fermentation parameters for CLA production were determined using a combination of single-factor and response surface methodologies: an inoculum size of 2%, a fermentation temperature of 36 °C, a fermentation time of 20 h, and a pine nut oil concentration of 11%. Under these optimized conditions, the resultant CLA yield was 33.47 µg/mL. Gas chromatography analysis revealed that the fermentation process yielded a mixture of c9, t11CLA and t10, c12 CLA isomers, representing 4.91% and 4.86% of the total fatty acid content, respectively.

Beta-blocker drives the conjugative transfer of multidrug resistance genes in pure and complex biological systems.

Drug resistance poses a high risk to human health. Extensive use of non-antibiotic drugs contributes to antibiotic resistance genes (ARGs) transfer. However, how they affect the spread of broad-host plasmids in complex biological systems remains unknown. This study investigated the effect of metoprolol on the transfer frequency and host range of ARGs in both intrageneric and intergeneric pure culture systems, as well as in anammox microbiome. The results showed that environmental concentrations of metoprolol significantly promoted the intrageneric and intergeneric conjugative transfer. Initially, metoprolol induced excessive oxidative stress, resulting in high cell membrane permeability and bacterial SOS response. Meanwhile, more pili formation increased the adhesion and contact between bacteria, and the abundance of conjugation-related genes also increased significantly. Activation of the electron transport chain provided more ATP for this energy-consuming process. The underlying mechanism was further verified in the complex anammox conjugative system. Metoprolol induced the enrichment of ARGs and mobile genetic elements. The enhanced bacterial interaction and energy generation facilitated the high conjugative transfer frequency of ARGs. In addition, plasmid-borne ARGs tended to transfer to opportunistic pathogens. This work raises public concerns about the health and ecological risks of non-antibiotic drugs.

One-pot Golden Gate Assembly of an avian infectious bronchitis virus reverse genetics system.

Avian infectious bronchitis is an acute respiratory disease of poultry of particular concern for global food security. Investigation of infectious bronchitis virus (IBV), the causative agent of avian infectious bronchitis, via reverse genetics enables deeper understanding of virus biology and a rapid response to emerging variants. Classic methods of reverse genetics for IBV can be time consuming, rely on recombination for the introduction of mutations, and, depending on the system, can be subject to genome instability and unreliable success rates. In this study, we have applied data-optimized Golden Gate Assembly design to create a rapidly executable, flexible, and faithful reverse genetics system for IBV. The IBV genome was divided into 12 fragments at high-fidelity fusion site breakpoints. All fragments were synthetically produced and propagated in E. coli plasmids, amenable to standard molecular biology techniques for DNA manipulation. The assembly can be carried out in a single reaction, with the products used directly in subsequent viral rescue steps. We demonstrate the use of this system for generation of point mutants and gene replacements. This Golden Gate Assembly-based reverse genetics system will enable rapid response to emerging variants of IBV, particularly important to vaccine development for controlling spread within poultry populations.

Heterostructured Mn-Sn Bimetallic Sulfide Nanocubes Confined in N, S-co-Doped Carbon Framework as High-Performance Anodes for Sodium-Ion Batteries.

Benefiting from its high theoretical capacity, tin disulfide (SnS2) draws abundant interest and attention for its promising practical prospect for sodium-ion batteries (SIBs). However, the huge volumetric variation in sodiation/desodiation reactions usually results in the fast decay of rate and cycling properties, which seriously obstructs its future applicable foregrounds. Herein, heterostructured Mn-Sn bimetallic sulfide nanocubes confined in N and S-codoped carbon (MSS@NSC) were rationally designed via a facile coprecipitation followed by a sulfurization strategy. When used as anodes for SIBs, the heterojunctions at the interfaces effectively accelerate the Na+ diffusion rate to promote the sodium-storage reaction kinetics. The N and S-codoped carbon provides a rapid conductive framework for the fast charge transport during the sodium-storage process. Moreover, the beneficial confinement effect of the NSC layer effectively guarantees a superb cycle property for the MSS@NSC anode. The favorable synergistic effects between the highly conductive framework of the NSC and MSS heterostructure endow the MSS@NSC anode with satisfactory electrochemical Na-storage properties.

Aminoselenation and Dehydroaromatization of Cyclohexanones with Anilines and Diselenides.

A three-component cascade reaction involving cyclohexanones, anilines, and diaryl diselenides under metal-free conditions is reported. The ortho-selenation of cyclohexanones with diaryl diselenides, followed by sequential dehydroaromatization with anilines, enables the preparation of a variety of o-selanyl anilines in moderate to excellent yields. This innovative transformation is notable for its excellent tolerance of functional groups and is suitable for the late-stage modification of complex pharmaceuticals.

Two-Phase Electrosynthesis of Dihydroxycoumestans: Discovery of a New Scaffold for Topoisomerase I Poison.

Coumestan represents a biologically relevant structural motif distributed in a number of natural products, and the rapid construction of related derivatives as well as the characterization of targets would accelerate lead compound discovery in medicinal chemistry. In this work, a general and scalable approach to 8,9-dihydroxycoumestans via two-electrode constant current electrolysis was developed. The application of a two-phase (aqueous/organic) system plays a crucial role for success, protecting the sensitive o-benzoquinone intermediates from over-oxidation. Based on the structurally diverse products, a primary SAR study on coumestan scaffold was completed, and compound 3 r exhibited potent antiproliferative activities and a robust topoisomerase I (Top1) inhibitory activity. Further mechanism studies demonstrates that compound 3 r was a novel Top1 poison, which might open an avenue for the development of Top1-targeted antitumor agent.

Pure-high-even-order dispersion bound solitons complexes in ultra-fast fiber lasers.

Temporal solitons have been the focus of much research due to their fascinating physical properties. These solitons can form bound states, which are fundamentally crucial modes in fiber laser and present striking analogies with their matter molecules counterparts, which means they have potential applications in large-capacity transmission and all-optical information storage. Although traditionally, second-order dispersion has been the dominant dispersion for conventional solitons, recent experimental and theoretical research has shown that pure-high-even-order dispersion (PHEOD) solitons with energy-width scaling can arise from the interaction of arbitrary negative-even-order dispersion and Kerr nonlinearity. Despite these advancements, research on the bound states of PHEOD solitons is currently non-existent. In this study, we obtained PHEOD bound solitons in a fiber laser using an intra-cavity spectral pulse shaper for high-order dispersion management. Specifically, we experimentally demonstrate the existence of PHEOD solitons and PHEOD bound solitons with pure-quartic, -sextic, -octic, and -decic dispersion. Numerical simulations corroborate these experimental observations. Furthermore, vibrating phase PHEOD bound soliton pairs, sliding phase PHEOD bound soliton pairs, and hybrid phase PHEOD bound tri-soliton are discovered and characterized. These results broaden the fundamental understanding of solitons and show the universality of multi-soliton patterns.

Elemental partitioning-mediated crystalline-to-amorphous phase transformation under quasi-static deformation.

The transformation induced plasticity phenomenon occurs when one phase transforms to another one during plastic deformation, which is usually diffusionless. Here we present elemental partitioning-mediated crystalline-to-amorphous phase transformation during quasi-static plastic deformation, in an alloy in form of a Cr-Ni-Co (crystalline)/Zr-Ti-Nb-Hf-Ni-Co (amorphous) nanolaminated composite, where the constitute elements of the two phases have large negative mixing enthalpy. Upon plastic deformation, atomic intermixing occurs between adjacent amorphous and crystalline phases due to extensive rearrangement of atoms at the interfaces. The large negative mixing enthalpy among the constituent elements promotes amorphous phase transformation of the original crystalline phase, which shows different composition and short-range-order structure compared with the other amorphous phase. The reduced size of the crystalline phase shortens mean-free-path of dislocations, facilitating strain hardening. The enthalpy-guided alloy design based on crystalline-to-amorphous phase transformation opens up an avenue for the development of crystal-glass composite alloys with ultrahigh strength and large plasticity.

Substantial removal of four pesticide residues in three fruits with ozone microbubbles.

Developing a straightforward method to remove pesticide residues from fruits is essential for food safety. In this study, ozone microbubble treatment was performed on three fruits (strawberry, cherry, and apricot) to remove four pesticide residues (emamectin benzoate, azoxystrobin, boscalid, and difenoconazole) while comparing removal efficiency. The concentration of hydroxyl radicals in different washing orientations was homogeneous at a concentration ranging between 8.9 and 10.2 µmol·L-1. Under long washing time (18 min), strawberry, cherry, and apricot obtained higher removal rates of 51 %∼65 %, 51 %∼59 % and 24 %∼70 %, respectively. Moreover, scanning electron microscopy (SEM) and contact angle (CA) revealed that apricot has better hydrophobicity, leading to a higher pesticide removal of 45 âˆ¼ 84 % with less water and more vigorous washing. Notably, vitamin C content in fruits remain largely unchanged following ozone microbubble treatment. This study demonstrated the effectiveness of ozone microbubble treatment as pollution-free method for enhancing food safety by removing pesticide residues on fruits.

Orthokeratology combined with spectacles in moderate to high myopia adolescents.

PURPOSE: Wearing ortho-k lenses overnight may not fully correct their daytime refractory errors of adolescents with moderate to high myopia. There are three common ways to deal with the daytime residual refractive error (RRE): 1) wearing spectacles to correct the RRE; 2) wear ortho-k lenses during the daytime instead of overnight.; 3) not correcting the residual refractive error. According to previous laboratory studies, myopic peripheral refraction is associated with better myopic control. This study had two aims:1) to compare relative peripheral refractive error (RPRE) among these ways after one-month stabilization; 2) to assess the axial length changes over 2 years of ortho-k lens overnight wear combined with spectacle glasses. METHODS: This was a prospective, non-controlled, non-randomized, observational study in which a total of 27 subjects (20 females, 7 males, mean age 12.48 ± 2.23Y) with spherical equivalent refractive error from -5.00 to -8.25D were enrolled. All participants in the study wore orthokeratology (ortho-k) lenses overnight for a minimum of one month. Subsequently, their peripheral refractive error (PRE) was assessed using an open-field autorefractor. During the assessment, the participants underwent three conditions in a random order in a same morning: 1) unaided eye after orthokeratology (referred to as the Unaided-eye condition), 2) wearing glasses to correct any remaining refractive errors after orthokeratology (referred to as the Spec-RE condition), and 3) wearing ortho-k lenses during the daytime (referred to as the Continuous OK wear condition). After testing, all subjects were instructed to wear ortho-k lenses overnight and glasses during the daytime to correct their RRE for the next 2 years, during which time the progression of their axial length was followed up. RESULTS: 1) RPRE in either Unaided-eye or Spec-RE condition subjects were significantly more myopic than those in the Continuous OK wear condition. 2) No difference in RPRE was seen between Unaided-eye and Spec-RE conditions. 3) Axial length growth was 0.05 ± 0.20 mm and 0.17 ± 0.32 mm (mean ± standard deviation) at 1-year and 2-year follow-ups after the initial visit, respectively, which were comparable to mild myopia patients after orthokeratology. 4) After orthokeratology, axial length change had negative correlation with the initial age (p = 0.001, r = -0.616) and residual diopter (p = 0.022). CONCLUSIONS: For myopes above refraction < -5.00D, wearing Ortho-k lenses overnight and glasses to correct the RRE in the daytime is recommended to ensure good visual quality and have more myopic RPRE for potential myopia control.

A juvenile mouse model of anti-N-methyl-D-aspartate receptor encephalitis by active immunization.

Introduction: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a common autoimmune encephalitis, and it is associated with psychosis, dyskinesia, and seizures. Anti-NMDAR encephalitis (NMDARE) in juveniles and adults presents different clinical charactreistics. However, the pathogenesis of juvenile anti-NMDAR encephalitis remains unclear, partly because of a lack of suitable animal models. Methods: We developed a model of juvenile anti-NMDAR encephalitis using active immunization with an amino terminal domain peptide from the GluN1 subunit (GluN1356 - 385) against NMDARs in 3-week-old female C57BL/6J mice. Results: Immunofluorescence staining suggested that autoantibody levels in the hippocampus increased, and HEK-293T cells staining identified the target of the autoantibodies as GluN1, suggesting that GluN1-specific immunoglobulin G was successfully induced. Behavior assessment showed that the mice suffered significant cognition impairment and sociability reduction, which is similar to what is observed in patients affected by anti-NMDAR encephalitis. The mice also exhibited impaired long-term potentiation in hippocampal CA1. Pilocarpine-induced epilepsy was more severe and had a longer duration, while no spontaneous seizures were observed. Conclusion: The juvenile mouse model for anti-NMDAR encephalitis is of great importance to investigate the pathological mechanism and therapeutic strategies for the disease, and could accelerate the study of autoimmune encephalitis.

Divergent Trideuteromethylthiolation and Aminotrideuteromethylthiolation of Alkenes with N-Fluorobenzenesulfonimide and CD3SSO3Na.

A metal-free oxidative trideuteromethylthiolation of alkenes with CD3SSO3Na using NFSI as an oxidant has been developed. The aminotrideuteromethylthiolation of a three-component reaction could be easily achieved in the presence of a cobalt catalyst. The divergent trideuteromethylthiolation of alkenes, readily available trideuteromethylthiolation reagent, and versatile synthetic strategy allow for late-stage modification of drug molecules.

Novel in-house knowledge-based automated planning system for lung cancer treated with intensity-modulated radiotherapy.

PURPOSE: The goal of this study was to propose a knowledge-based planning system which could automatically design plans for lung cancer patients treated with intensity-modulated radiotherapy (IMRT). METHODS AND MATERIALS: From May 2018 to June 2020, 612 IMRT treatment plans of lung cancer patients were retrospectively selected to construct a planning database. Knowledge-based planning (KBP) architecture named αDiar was proposed in this study. It consisted of two parts separated by a firewall. One was the in-hospital workstation, and the other was the search engine in the cloud. Based on our previous study, A­Net in the in-hospital workstation was used to generate predicted virtual dose images. A search engine including a three-dimensional convolutional neural network (3D CNN) was constructed to derive the feature vectors of dose images. By comparing the similarity of the features between virtual dose images and the clinical dose images in the database, the most similar feature was found. The optimization parameters (OPs) of the treatment plan corresponding to the most similar feature were assigned to the new plan, and the design of a new treatment plan was automatically completed. After αDiar was developed, we performed two studies. The first retrospective study was conducted to validate whether this architecture was qualified for clinical practice and involved 96 patients. The second comparative study was performed to investigate whether αDiar could assist dosimetrists in improving the quality of planning for the patients. Two dosimetrists were involved and designed plans for only one trial with and without αDiar; 26 patients were involved in this study. RESULTS: The first study showed that about 54% (52/96) of the automatically generated plans would achieve the dosimetric constraints of the Radiation Therapy Oncology Group (RTOG) and about 93% (89/96) of the automatically generated plans would achieve the dosimetric constraints of the National Comprehensive Cancer Network (NCCN). The second study showed that the quality of treatment planning designed by junior dosimetrists was improved with the help of αDiar. CONCLUSIONS: Our results showed that αDiar was an effective tool to improve planning quality. Over half of the patients' plans could be designed automatically. For the remaining patients, although the automatically designed plans did not fully meet the clinical requirements, their quality was also better than that of manual plans.

A Novel Al-Cu Composite with Ultra-High Strength at 350 °C via Dual-Phase Particle Reinforced Submicron-Structure.

Thermal stability determines a material's ability to maintain its performance at desired service temperatures. This is especially important for aluminum (Al) alloys, which are widely used in the commercial sector. Herein, an ultra-strong and heat-resistant Al-Cu composite is fabricated with a structure of nano-AlN and submicron-Al2 O3 particles uniformly distributed in the matrix. At 350 °C, the (8.2AlN+1Al2 O3 )p /Al-0.9Cu composite achieves a high strength of 187 MPa along with a 4.6% ductility under tension. The high strength and good ductility benefit from strong pinning effect on dislocation motion and grain boundary sliding by uniform dispersion of nano-AlN particles, in conjunction with the precipitation of Guinier-Preston (GP) zones, enhancing strain hardening capacity during plastic deformation. This work can expand the selection of Al-Cu composites for potential applications at service temperatures as high as ≈350 °C.

Both epilepsy and anti-seizure medications affect bone metabolism in children with self-limited epilepsy with centrotemporal spikes.

OBJECTIVE: Bone metabolism can be influenced by a range of factors. We selected children with self-limited epilepsy with centrotemporal spikes (SeLECTS) and lifestyles similar to those of healthy children to control for the confounding factors that may influence bone metabolism. We aimed to identify the specific effects of epilepsy and/or anti-seizure medications (ASMs) on bone metabolism. METHODS: Patients with SeLECTS were divided into an untreated group and a monotherapy group, and the third group was a healthy control group. We determined the levels of various biochemical markers of bone metabolism, including procollagen type I nitrogenous propeptide (PINP), alkaline phosphatase (ALP), osteocalcin (OC), collagen type I cross-linked C-telopeptide (CTX), calcium, magnesium, phosphorus, parathyroid hormone (PTH), and vitamin D3 (VD3 ). RESULTS: A total of 1487 patients (from 19 centers) were diagnosed with SeLECTS; 1032 were analyzed, including 117 patients who did not receive any ASMs (untreated group), 643 patients who received only one ASM (monotherapy group), and 272 children in the healthy control group. Except for VD3 , other bone metabolism of the three groups were different (p < .001). Bone metabolism was significantly lower in the untreated group than the healthy control group (p < .05). There were significant differences between the monotherapy and healthy control group in the level of many markers. However, when comparing the monotherapy and untreated groups, the results were different; oxcarbazepine, levetiracetam, and topiramate had no significant effect on bone metabolism. Phosphorus and magnesium were significantly lower in the valproic acid group than the untreated group (adjusted p < .05, Cliff's delta .282-.768). CTX was significantly higher in the lamotrigine group than in the untreated group (adjusted p = .012, Cliff's delta = .316). SIGNIFICANCE: Epilepsy can affect many aspects of bone metabolism. After controlling epilepsy and other confounders that affect bone metabolism, we found that the effects of ASMs on bone metabolism differed. Oxcarbazepine, levetiracetam, and topiramate did not affect bone metabolism, and lamotrigine corrected some of the abnormal markers of bone metabolism in patients with epilepsy.

Alkyltin fluorides as alkylating reagent in aminoalkylation of maleimides.

Alkyltin fluoride is a frequently used electrophilic stannylation reagent via the cleavage of the Sn-F bond in traditional organic synthetic chemistry. Herein, we report the unprecedented copper-catalyzed aminoalkylation of maleimides using alkyltin fluoride as alkylating reagent through cleavage of the C-Sn bond via a radical pathway. Excellent functional group tolerance, use of O2 as green oxidant and late-stage modification of some drug intermediates are the standout features of the current toolbox. Mechanistic studies reveal that alkyltin fluorides are capable of producing alkyl radicals in a Cu/O2 catalytic system.

Palladium-Catalyzed N-Alkenylation of N-Aryl Phosphoramidates with Alkenes.

Versatile and concise Pd-catalyzed oxidative N-alkenylation of N-aryl phosphoramidates with alkenes is described in this study, a reaction that is of great significance but surprisingly unexploited. The transformation proceeds under mild reaction conditions, using O2 as a green oxidant and TBAB as an effective additive. An efficient catalytic system allows a variety of drug-related substrates to participate in these transformations, which is of great interest in the drug discovery and development of phosphoramidates.