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Rechargeable magnesium metal batteries (RMBs) have shown promising prospects in sustainable energy storage due to the high crustal abundance, safety, and potentially large specific capacity of magnesium. However, their development is constrained by the lack of effective cathode materials that can achieve high capacity and stable magnesium storage at a practically reasonable rate. Herein, we construct a three-dimensional (3D) iron(III)-dihydroxy-benzoquinone (Fe2(DHBQ)3) metal-organic framework (MOF) material with dual redox centers of Fe3+ cations and DHBQ2- anions for reversible storage of Mg2+ in RMBs. Spectroscopic analysis and density functional theory (DFT) calculations reveal the redox chemistry of both Fe3+ ions and carbonyls from DHBQ ligands during electrochemical processes. Benefiting from the rational structure, the Fe2(DHBQ)3â¥Mg cells exhibit a high reversible capacity of 395.3 mAh/g, large energy density of 463.5 Wh/kg, and high power density of 2456.0 W/kg. Moreover, the high electronic conductivity (8.35 × 10-5 S/cm) and favorable diffusion path of Mg2+ in Fe2(DHBQ)3 endow the cells with exceptional cycling stability and rate capability with a long life of 5000 cycles at 2000 mA/g. The dual redox-active MOF demonstrates a category of advanced cathode materials for high-performance RMBs.
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Structural topology and symmetry of a two-dimensional (2D) network play pivotal roles in defining its electrical properties and functionalities. Here, a binary buckled honeycomb lattice with C3v symmetry, which naturally hosts topological Dirac fermions and out-of-plane polarity, is proposed. It is successfully achieved in a group IV-V compound, namely monolayer SiP epitaxially grown on Ag(111) surface. Combining first-principles calculations with angle-resolved photoemission spectroscopy, the degeneration of the Dirac nodal lines to points due to the broken horizonal mirror symmetry is elucidated. More interesting, the SiP monolayer manifests metallic nature, which is mutually exclusive with polarity in conventional materials. It is further found that the out-of-plane polarity is strongly suppressed by the metallic substrate. This study not only represents a breakthrough of realizing intrinsic polarity in 2D metallic material via ingenious design but also provides a comprehensive understanding of the intricate interplay of many exotic low-dimensional quantum phenomena.
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Polymer electrolytes play a crucial role in advancing rechargeable magnesium batteries (RMBs) owing to their exceptional characteristics, including high flexibility, superior interface compatibility, broad electrochemical stability window, and enhanced safety features. Despite these advantages, research in this domain remains nascent, plagued by single preparation approaches and challenges associated with the compatibility between polymer electrolytes and Mg metal anode. In this study, we present a novel synthesis strategy to fabricate a glycerol α,α'-diallyl ether-3,6-dioxa-1,8-octanedithiol-based composite gel polymer electrolyte supported by glass fiber substrate (GDT@GF CGPE) through anion modification and thiol-ene click chemistry polymerization. The developed route exhibits novelty and high efficiency, leading to the production of GDT@GF CGPEs featuring exceptional mechanical properties, heightened ionic conductivity, elevated Mg2+ transference number, and commendable compatibility with Mg anode. The assembled modified Mo6S8||GDT@GF||Mg cells exhibit outstanding performance across a wide temperature range and address critical safety concerns, showcasing the potential for applications under extreme conditions. Our innovative preparation strategy offers a promising avenue for the advancement of polymer electrolytes in high-performance rechargeable magnesium batteries, while also opens up possibilities for future large-scale applications and the development of flexible electronic devices.
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Colloidal nanoparticles offer unique photoelectric properties, making them promising for functional applications. Multiparticle systems exhibit synergistic effects on the functional properties of their individual components. However, precisely controlled assembly of multiparticles to form patterned building blocks for solid-state devices remains challenging. Here, we demonstrate a versatile multiparticle synergistic electrophoretic deposition (EPD) strategy to achieve controlled assembly, high-efficiency, and high-resolution patterns. Through elaborate surface design and charge regulation of nanoparticles, we achieve precise control over the particle distribution (gradient or homogeneous structure) in multiparticle films using the EPD technique. The multiparticle system integrates silicon oxide and titanium oxide nanoparticles, synergistically enhancing the emission efficiency of quantum dots to a high level in the field. Furthermore, we demonstrate the superiority of our strategy to integrate multiparticle into large-area full-color display panels with a high resolution over 1000 pixels per inch. The results suggest great potential for developing multiparticle systems and expanding diverse functional applications.
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The complex interrelationships among thermoelectric parameters mean that a priori design of high-performing materials is difficult. However, band engineering can allow the power factor to be optimized through enhancement of the Seebeck coefficient. Herein, using layered Sb2Si2Te6 and Sc2Si2Te6 as model systems, we comprehensively investigate and compare their thermoelectric properties by employing density functional theory combined with semiclassical Boltzmann transport theory. Our simulations reveal that Sb2Si2Te6 exhibits superior electrical conductivity compared to Sc2Si2Te6 due to lower scattering rates and more pronounced band dispersion. Remarkably, despite Sb2Si2Te6 exhibiting a lower lattice thermal conductivity and superior electrical conductivity, Sc2Si2Te6 is predicted to achieve an extraordinary dimensionless figure of merit (ZT) of 3.51 at 1000 K, which significantly surpasses the predicted maximum ZT of 2.76 for Sb2Si2Te6 at 900 K. We find the origin of this behavior to be a combined increase in band (valley) degeneracy and anisotropy upon switching the conduction band orbital character from Sb p to Sc d, yielding a significantly improved Seebeck coefficient. This work suggests that enhancing band degeneracy and anisotropy (complexity) through compositional variation is an effective strategy for improving the thermoelectric performance of layered materials.
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BACKGROUND: The therapeutic impact of the Wenyang Huoxue (WYXH) formula on coronary atherosclerotic heart disease (CHD) is well established, yet the precise mechanisms are currently not fully understood. This study provides preliminary insights into the potential mechanisms underlying the therapeutic effects of the formula on CHD by utilizing network pharmacology and molecular docking technology. MATERIALS AND METHODS: The primary active constituents and their corresponding action targets for the formula were retrieved from the TCMSP database. Utilizing Cytoscape 3.9.1 software, a network linking the components of the formula to their respective targets was constructed. Information was collected from Genecards, OMIM, TTD, and DrugBank databases to identify targets related to CHD. The common targets shared by the formula and CHD were then imported into the STRING database to create a protein-protein interaction (PPI) network. Following this, enrichment analyses were performed on the shared targets using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Finally, molecular docking was conducted on the primary active compounds and the core targets. RESULTS: The network encompassing the components and targets of the formula comprises a total of 311 nodes and 895 edges. Compounds exhibiting higher degree centrality consist of quercetin, ß-sitosterol, and kaempferol. In the PPI network, proteins with elevated degree centrality are protein kinase B (AKT1), epidermal growth factor receptor (EGFR), and mitogen-activated protein kinase 3 (MAPK3). The results of GO and KEGG enrichment analyses reveal that the biological processes associated with the efficacy of the formula in treating CHD primarily involve positive regulation of gene expression, hypoxia response, and lipopolysaccharide response, among others. The signaling pathways primarily involved include phosphatidylinositol 3-kinase and protein kinase B (PI3K-AKT), MAPK3, tumor necrosis factor (TNF), and so on. Molecular docking results demonstrate a strong affinity between quercetin, ß-sitosterol, and kaempferol with AKT1, EGFR, and MAPK3. CONCLUSION: We showed for the first time that AKT1, EGFR, and MAPK3 are potential targets influenced by the WYHX formula in CHD treatment. The therapeutic effects could possibly involve signaling pathways such as the PI3K-AKT, MAPK, TNF, and AGE-RAGE pathways.
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Maladie des artères coronaires , Médicaments issus de plantes chinoises , Simulation de docking moléculaire , Pharmacologie des réseaux , Cartes d'interactions protéiques , Médicaments issus de plantes chinoises/pharmacologie , Médicaments issus de plantes chinoises/composition chimique , Médicaments issus de plantes chinoises/usage thérapeutique , Humains , Maladie des artères coronaires/traitement médicamenteux , Sitostérol/pharmacologie , Transduction du signal/effets des médicaments et des substances chimiques , Protéines proto-oncogènes c-akt/métabolismeRÉSUMÉ
[This corrects the article DOI: 10.3389/fnut.2022.1024722.].
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OBJECTIVE: This paper was aimed at unveiling the effect of low-molecular-weight heparin calcium (LMWH) combined with magnesium sulfate and labetalol on coagulation, vascular endothelial function, and pregnancy outcome in early-onset severe preeclampsia (EOSP). METHODS: Pregnant women with EOSP were divided into the control group and the study group, each with 62 cases. Patients in the control group were treated with labetalol and magnesium sulfate, and those in the study group were treated with LMWH in combination with the control grou Blood pressure (systolic blood pressure [SBP] and diastolic blood pressure [DBP]), 24-h urine protein, coagulation indices [D-dimer (D-D), plasma fibrinogen (Fg), prothrombin time (PT), activated partial thromboplastin time (APTT), and prothrombin time (TT)], endothelial function [endothelin (ET-1) and nitric oxide (NO)], oxidative stress indices [oxidized low-density lipoproteins (ox-LDL), lipid peroxidation (LPO), superoxide dismutase (SOD), and malondialdehyde (MDA)], pregnancy outcome, and adverse effects occurred in the two groups were compared. RESULTS: After treatment, lower SBP, DBP, and 24-h urine protein levels; lower Fg and D-D levels; higher PT, APPT, and TT levels; higher NO levels; lower ET-1 levels; lower ox-LDL, MDA, and LPO levels; higher SOD levels; and lower incidence of adverse pregnancy and adverse reactions were noted in the study group in contrast to the control group. CONCLUSION: EOSP patients given with LMWH combined with magnesium sulfate and labetalol can effectively reduce the patient's blood pressure and urinary protein level; improve coagulation function, oxidative stress, and vascular endothelial function indices; reduce the adverse pregnancy outcomes; and improve the safety of treatment.
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Metabolic remodeling is a strategy for tumor survival under stress. However, the molecular mechanisms during the metabolic remodeling of colorectal cancer (CRC) remain unclear. Melanocyte proliferating gene 1 (MYG1) is a 3'-5' RNA exonuclease and plays a key role in mitochondrial functions. Here, we uncover that MYG1 expression is upregulated in CRC progression and highly expressed MYG1 promotes glycolysis and CRC progression independent of its exonuclease activity. Mechanistically, nuclear MYG1 recruits HSP90/GSK3ß complex to promote PKM2 phosphorylation, increasing its stability. PKM2 transcriptionally activates MYC and promotes MYC-medicated glycolysis. Conversely, c-Myc also transcriptionally upregulates MYG1, driving the progression of CRC. Meanwhile, mitochondrial MYG1 on the one hand inhibits oxidative phosphorylation (OXPHOS), and on the other hand blocks the release of Cyt c from mitochondria and inhibits cell apoptosis. Clinically, patients with KRAS mutation show high expression of MYG1, indicating a high level of glycolysis and a poor prognosis. Targeting MYG1 may disturb metabolic balance of CRC and serve as a potential target for the diagnosis and treatment of CRC.
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Tumeurs colorectales , Glycolyse , Mitochondries , Phosphorylation oxydative , Animaux , Femelle , Humains , Mâle , Souris , Apoptose/génétique , Protéines de transport/métabolisme , Protéines de transport/génétique , Lignée cellulaire tumorale , Noyau de la cellule/métabolisme , Tumeurs colorectales/métabolisme , Tumeurs colorectales/génétique , Tumeurs colorectales/anatomopathologie , Régulation de l'expression des gènes tumoraux , Glycogen synthase kinase 3 beta/métabolisme , Glycogen synthase kinase 3 beta/génétique , Protéines du choc thermique HSP90/métabolisme , Protéines du choc thermique HSP90/génétique , Protéines membranaires/métabolisme , Protéines membranaires/génétique , Souris nude , Mitochondries/métabolisme , Protéines proto-oncogènes c-myc/métabolisme , Protéines proto-oncogènes c-myc/génétique , Protéines proto-oncogènes p21(ras)/métabolisme , Protéines proto-oncogènes p21(ras)/génétique , , Hormones thyroïdiennes/métabolisme , Hormones thyroïdiennes/génétiqueRÉSUMÉ
Patients with chronic pelvic inflammation (CPI) experience irregular menstrual, ectopic pregnancy, and infertility. Yiyi Baijiang Decoction attenuates CPI in patients with uncovered mechanisms. CPI therapeutic targets intersected with those of Yiyi Baijiang Decoction, followed by importing into STRING to obtain protein-target interaction. "Drug-component-disease-target" interaction was constructed by Cytoscape. mRNA and protein levels were detected by real-time quantitative PCR (RT-qPCR) and western blot. Yiyi Baijiang Decoction contained 199 active ingredients. There were 1071 drug targets for Yiyi Baijiang Decoction and 1622 therapeutic targets for CPI. The GO functional enrichment analysis revealed 3445 biological processes, and the KEGG pathway enrichment analysis screened 67 signal pathways. Decreased ALB, increased protein kinase B (AKT1), interleukin (IL)-6, vascular endothelial growth factor A (VEGFA), and phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K/AKT)-extracellular-regulated protein kinases (ERK)1/2 activation in CPI mice were abolished by Yiyi Baijiang Decoction. Yiyi Baijiang Decoction attenuates CPI by inactivating PI3K/AKT and ERK1/2 and regulating ALB, VEGFA, AKT1, and IL-6.
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Flavonoids are widely distributed in nature and have a variety of beneficial biological effects, including antioxidant, anti-inflammatory, and anti-obesity effects. All of these are related to gut microbiota, and flavonoids also serve as a bridge between the host and gut microbiota. Flavonoids are commonly used to modify the composition of the gut microbiota by promoting or inhibiting specific microbial species within the gut, as well as modifying their metabolites. In turn, the gut microbiota extensively metabolizes flavonoids. Hence, this reciprocal relationship between flavonoids and the gut microbiota may play a crucial role in maintaining the balance and functionality of the metabolism system. In this review, we mainly highlighted the biological effects of antioxidant, anti-inflammatory and antiobesity, and discussed the interaction between flavonoids, gut microbiota and lipid metabolism, and elaborated the potential mechanisms on host lipid metabolism.
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The advent of 2D materials has ushered in the exploration of their synthesis, characterization and application. While plenty of 2D materials have been synthesized on various metallic substrates, interfacial interaction significantly affects their intrinsic electronic properties. Additionally, the complex transfer process presents further challenges. In this context, experimental efforts are devoted to the direct growth on technologically important semiconductor/insulator substrates. This review aims to uncover the effects of substrate on the growth of 2D materials. The focus is on non-metallic substrate used for epitaxial growth and how this highlights the necessity for phase engineering and advanced characterization at atomic scale. Special attention is paid to monoelemental 2D structures with topological properties. The conclusion is drawn through a discussion of the requirements for integrating 2D materials with current semiconductor-based technology and the unique properties of heterostructures based on 2D materials. Overall, this review describes how 2D materials can be fabricated directly on non-metallic substrates and the exploration of growth mechanism at atomic scale.
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A new peptide-based fluorescent probe named DMDH with easy-to-synthesize, excellent stability, good water solubility and large Stokes shift (225 nm) was synthesized for highly selective sequential detections of copper ions (Cu2+) and glyphosate (Glyp). DMDH demonstrated great detection performance towards Cu2+via strong fluorescence quenching, and forming non-fluorescence DMDH-Cu2+ ensemble. As a new promising cascade probe, the fluorescence of DMDH-Cu2+ ensemble was significantly recovered based on displacement approach after glyphosate was added. Interestingly, the limit of detections (LODs) for Cu2+ and glyphosate were 40.6 nM and 10.6 nM, respectively, which were far lower than those recommended by the WHO guidelines for drinking water. More importantly, DMDH was utilized to evaluate Cu2+ and glyphosate content in three real water samples, demonstrating that its effectiveness in water quality monitoring. Additionally, it is worth noting that DMDH was also applied to analyze Cu2+ and glyphosate in living cells in view of significant cells permeability and low cytotoxicity. Moreover, DMDH soaked in filter paper was used to create qualitative test strips and visually identify Cu2+ and glyphosate through significant color changes. Furthermore, smartphone RGB color recognition provided a new method for semi-quantitative testing of Cu2+ and glyphosate in the absence of expensive instruments.
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Cuivre , Colorants fluorescents , Glycine , , Peptides , Ordiphone , Spectrométrie de fluorescence , Cuivre/analyse , Cuivre/composition chimique , Glycine/analogues et dérivés , Glycine/analyse , Glycine/composition chimique , Colorants fluorescents/composition chimique , Colorants fluorescents/synthèse chimique , Humains , Spectrométrie de fluorescence/méthodes , Peptides/composition chimique , Limite de détection , Bandelettes réactives/analyse , Polluants chimiques de l'eau/analyse , Cellules HeLa , Eau de boisson/analyseRÉSUMÉ
Mercury ions (Hg2+) and sulfur ions (S2-), have caused serious harm to the ecological environment and human health as two kinds of highly toxic pollutants widely used. Therefore, the visual quantitative determination of Hg2+ and S2- is of great significance in the field of environmental monitoring and medical therapy. In this study, a novel fluorescent "on-off-on" peptide-based probe DNC was designed and synthesized using dipeptide (Asn-Cys-NH2) as the raw material via solid phase peptide synthesis (SPPS) technology with Fmoc chemistry. DNC displayed high selectivity in the recognition of Hg2+, and formed non-fluorescence complex (DNC-Hg2+) through 2:1 binding mode. Notably, DNC-Hg2+ complex generated in situ was used as relay response probe for highly selective sequential detection of S2- through reversible formation-separation. DNC achieved highly sensitive detection of Hg2+ and S2- with the detection limits (LODs) of 8.4 nM and 5.5 nM, respectively. Meanwhile, DNC demonstrated feasibility for Hg2+ and S2- detections in two water samples, and the considerable recovery rate was obtained. More importantly, DNC showed excellent water solubility and low toxicity, and was successfully used for consecutive discerning Hg2+ and S2- in test strips, living cells and zebrafish larvae. As an effective visual analysis method in the field, smartphone RGB Color Picker APP realized semi-quantitative detections of Hg2+ and S2- without the need for complicated device.
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Colorants fluorescents , Mercure , Peptides , Danio zébré , Mercure/analyse , Animaux , Colorants fluorescents/composition chimique , Colorants fluorescents/synthèse chimique , Humains , Peptides/composition chimique , Peptides/analyse , Spectrométrie de fluorescence , Limite de détection , Soufre/composition chimique , Soufre/analyse , Polluants chimiques de l'eau/analyse , Imagerie optique , Cellules HeLa , Ions/analyseRÉSUMÉ
The excessive emission of copper ions (Cu2+) and the abuse of glyphosate (Glyp) have caused serious harm to the ecological environment and human health, so it is important to develop a fast and convenient method for the analysis of Cu2+ and glyphosate to ensure environmental and food safety. Herein, a dual-signals peptide-based probe (FASRH) with fluorescent and colorimetric was prepared using 5-carboxyl fluorescein modified tetrapeptide (Ala-Ser-Arg-His-NH2). FASRH was successfully used to recognize Cu2+ as a fluorescence "on-off" probe, forming the FASRH-Cu2+ complex with non-fluorescence. As a new promising cascade probe, FASRH-Cu2+ complex probe has high selectivity (only Glyp), good sensitivity (50.2 nM), good anti-interference ability and wide pH range (7.0-11.0) for the detection of glyphosate by ligand replacement method. In addition, the recognizable color changed markedly under 365 nm UV light and natural light. Notably, FASRH not only achieved accurate monitoring of Cu2+ and glyphosate in two real water samples, but also successfully applied to detect Cu2+ and glyphosate in live Hacat cells based on low cytotoxicity. Moreover, it is worth noting that FASRH-impregnated test strips exhibited significant fluorescence and colorimetric color changes for Cu2+ and glyphosate via naked eye. Furthermore, smartphone-assisted FASRH was used for the portable detection of Cu2+ and glyphosate based on the advantages of simplicity, low cost and fast response.
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Colorimétrie , Cuivre , Colorants fluorescents , Glycine , , Spectrométrie de fluorescence , Glycine/analogues et dérivés , Glycine/analyse , Cuivre/analyse , Humains , Colorimétrie/méthodes , Colorants fluorescents/composition chimique , Lignée cellulaire , Polluants chimiques de l'eau/analyse , Peptides/composition chimiqueRÉSUMÉ
In this work, a new ratiometric fluorescent probe DKA was synthesized based on the double sides of lysine backbone conjugated with alanine and dansyl groups. DKA exhibited fluorescence ratiometric response for Hg2+ with high sensitivity (13.4 nM), specific selectivity (only Hg2+), strong anti-interference ability (no interference), fast recognition (within 60 s) and wide pH range (5-10). The stoichiometry of binding of DKA and Hg2+ was determined to be 1:1 via Job's plot, ESI-HRMS and 1HNMR titration analysis. Subsequently, the in situ formation of DKA-Hg2+ complex was used for highly selective detection of S2- as a novel fluorescence "on-off" probe, and the lowest detection limit for S2- was 12.9 nM. In addition, DKA possessed excellent cells permeation and low toxicity, and fluorescence imaging of Hg2+ and S2- was performed in living Hacat cells. Most importantly, the digital imaging using a smartphone color recognition APP indicated that DKA could semi-quantitatively and visually detected Hg2+ and S2- without expensive equipment.
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Colorants fluorescents , Mercure , Ordiphone , Spectrométrie de fluorescence , Mercure/analyse , Colorants fluorescents/composition chimique , Colorants fluorescents/synthèse chimique , Humains , Peptides/composition chimique , Peptides/synthèse chimique , Limite de détection , Lignée cellulaire , Imagerie optique , Concentration en ions d'hydrogèneRÉSUMÉ
Age structure is one of the crucial factors in characterizing the heterogeneous epidemic transmission. Vaccination is regarded as an effective control measure for prevention and control epidemics. Due to the shortage of vaccine capacity during the outbreak of epidemics, how to design vaccination policy has become an urgent issue in suppressing the disease transmission. In this paper, we make an effort to propose an age-structured SVEIHR model with the disease-caused death to take account of dynamics of age-related vaccination policy for better understanding disease spread and control. We present an explicit expression of the basic reproduction number R 0 , which determines whether or not the disease persists, and then establish the existence and stability of endemic equilibria under certain conditions. Numerical simulations are illustrated to show that the age-related vaccination policy has a tremendous influence on curbing the disease transmission. Especially, vaccination of people over 65 is better than for people aged 21-65 in terms of rapid eradication of the disease in Italy.
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Épidémies , Vaccination , Humains , Épidémies de maladies/prévention et contrôle , Taux de reproduction de base , Épidémies/prévention et contrôle , ItalieRÉSUMÉ
Gastric cancer is one of the cancers with increasing incidence and ranks fourth globally among the most frequent causes of cancer-related mortality. Early gastric cancer is often asymptomatic or presents with atypical symptoms, and the majority of patients present with advanced disease upon diagnosis. Brain metastases are present in approximately 1% of gastric cancer patients at the time of diagnosis, which significantly contributed to the overall mortality of the disease worldwide. Conventional therapies for patients with brain metastases remain limited and the median overall survival of patients is only 8 months in advanced cases. Recent studies have improved our understanding of the molecular mechanisms underlying gastric cancer brain metastases, and immunotherapy has become an important treatment option in combination with radiotherapy, chemotherapy, targeted therapy and surgery. This review aims to provide insight into the cellular processes involved in gastric cancer brain metastases, discuss diagnostic approaches, evaluate the integration of immune checkpoint inhibitors into treatment and prognosis, and explore the predictive value of biomarkers in immunotherapy.
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Infertility can be stressful for infertile couples. This study aims to examine the intra-dyadic associations between stigma, communication patterns, and infertility-related stress in couples undergoing artificial insemination by donor semen (AID). This cross-sectional study was conducted from January to April 2021. Two hundred and three couples undergoing AID were recruited from a reproductive centre in China. All of the couples completed a two-item stigma questionnaire, Communication Pattern Questionnaire, and Fertility Problem Inventory. The actor-partner interdependence mediation analysis was performed using AMOS 23.0. The analysis demonstrated significant actor-actor effects for couples undergoing AID. More specifically, higher levels of stigma among wives and husbands were associated with more negative communication patterns, thereby increasing their own infertility-related stress. Simultaneously, there was a significant partner-actor effect among infertile wives, demonstrating that the husband's stigma can affect his wife's infertility-related stress by influencing her communication patterns. Couples undergoing AID experience increased infertility-related stress when they have high levels of stigma and negative communication patterns, and husbands' stigma is correlated to wives' communication patterns. Therefore, dyadic interventions aiming to improving stigma and enhancing positive communication may be conducive to reducing infertility-related stress.