Molecular Subtypes Defined by Cuproptosis-Associated Genes, Prognostic Model Development, and Tumor Immune Microenvironment Characterization in Adrenocortical Carcinoma.

Introduction: This study aims to explore the role of cuproptosis-related genes in ACC, utilizing data from TCGA and GEO repositories, and to develop a predictive model for patient stratification. Methods: A cohort of 123 ACC patients with survival data was analyzed. RNA-seq data of 17 CRGs were examined, and univariate Cox regression identified prognostic CRGs. A cuproptosis-related network was constructed to show interactions between CRGs. Consensus clustering classified ACC into three subtypes, with transcriptional and survival differences assessed by PCA and survival analysis. Gene set variation analysis (GSVA) and ssGSEA evaluated functional and immune infiltration characteristics across subtypes. Differentially expressed genes (DEGs) were identified, and gene clusters were established. A risk score (CRG_score) was generated using LASSO and multivariate Cox regression, validated across datasets. Tumor microenvironment, stem cell index, mutation status, drug sensitivity, and hormone synthesis were examined in relation to the CRG_score. Protein expression of key genes was validated, and functional studies on ASF1B and NDRG4 were performed. Results: Three ACC subtypes were identified with distinct survival outcomes. Subtype B showed the worst prognosis, while subtype C had the best. We identified 214 DEGs linked to cell proliferation and classified patients into three gene clusters, confirming their prognostic value. The CRG_score predicted patient outcomes, with high-risk patients demonstrating worse survival and possible resistance to immunotherapy. Drug sensitivity analysis suggested higher responsiveness to doxorubicin and etoposide in high-risk patients. Conclusion: This study suggests the potential prognostic value of CRGs in ACC. The CRG_score model provides a robust tool for risk stratification, with implications for treatment strategies.

Disulfidptosis-related subtype and prognostic signature in prostate cancer.

BACKGROUND: Disulfidptosis refers to cell death caused by the accumulation and bonding of disulfide in the cytoskeleton protein of SLC7A11-high level cells under glucose deprivation. However, the role of disulfidptosis-related genes (DRGs) in prostate cancer (PCa) classification and regulation of the tumor microenvironment remains unclear. METHODS: Firstly, we analyzed the expression and mutation landscape of DRGs in PCa. We observed the expression levels of SLC7A11 in PCa cells through in vitro experiments and assessed the inhibitory effect of the glucose transporter inhibitor BAY-876 on SLC7A11-high cells using CCK-8 assay. Subsequently, we performed unsupervised clustering of the PCa population and analyzed the differentially expressed genes (DEGs) between clusters. Using machine learning techniques to select a minimal gene set and developed disulfidoptosis-related risk signatures for PCa. We analyzed the tumor immune microenvironment and the sensitivity to immunotherapy in different risk groups. Finally, we validated the accuracy of the prognostic signatures genes using single-cell sequencing, qPCR, and western blot. RESULTS: Although SLC7A11 can increase the migration ability of tumor cells, BAY-876 effectively suppressed the viability of prostate cancer cells, particularly those with high SLC7A11 expression. Based on the DRGs, PCa patients were categorized into two clusters (A and B). The risk label, consisting of a minimal gene set derived from DEGs, included four genes. The expression levels of these genes in PCa were initially validated through in vitro experiments, and the accuracy of the risk label was confirmed in an external dataset. Cluster-B exhibited higher expression levels of DRG, representing lower risk, better prognosis, higher immune cell infiltration, and greater sensitivity to immune checkpoint blockade, whereas Cluster A showed the opposite results. These findings suggest that DRGs may serve as targets for PCa classification and treatment. Additionally, we constructed a nomogram that incorporates DRGs and clinical pathological features, providing clinicians with a quantitative method to assess the prognosis of PCa patients. CONCLUSION: This study analyzed the potential connection between disulfidptosis and PCa, and established a prognostic model related to disulfidptosis, which holds promise as a valuable tool for the management and treatment of PCa patients.

A Cascade Activation Probe with Double-Enhanced Near-Infrared Imaging for Monitoring Peroxynitrite Fluctuations in Vivo.

Monitoring peroxynitrite (ONOO-) fluctuations is particularly important for assessing pathological progression and oxidative damage due to their crucial role in maintaining the redox balance of organisms. However, due to the lack of efficient tools for differentially monitoring ONOO- fluctuations at different concentration ranges in vivo, the precise detection of endogenous ONOO- fluctuations under pathological conditions in living systems remains challenging. Herein, we rationally designed a double-enhanced emission cascade activatable near-infrared (NIR) fluorescent probe (B-TCF) for the measurement of ONOO-, which consists of a borate ester response group and a malononitrile hemicyanine fluorophore. Especially, after sequential oxidative hydrolysis of the borate ester group and xanthene skeleton, B-TCF exhibited a sequentially double-enhanced NIR emission response at 776 and 625 nm for different ONOO- concentration ranges. Moreover, B-TCF revealed excellent and promising performance for ONOO- in terms of high selectivity, sensitivity, and reaction rate (k = 28.2 M-1 s-1). Motivated by the two-step emission signal enhancement and large wavelength shift in the NIR region, B-TCF enabled discriminative imaging of ONOO- with the low and high concentrations in living cells. Importantly, B-TCF was successfully applied for assessing the pathological progression of isoniazid and acetaminophen-induced liver damage in vivo by detecting the endogenous different ONOO- levels. Overall, this study not only demonstrates the first double-enhanced emission cascade activatable NIR fluorescent probe for measuring the dynamic variation of ONOO- in related diseases but also shows great potential as an effective molecular tool for evaluating the various stages of drug-induced liver damage.

Enhancement of the flavor and functional characteristics of cod protein isolate using an enzyme-microbe system.

Cod protein isolate (CPI), a by-product of the cod processing industry, represents a novel source of high value-added products. However, off-flavors in cod protein such as bitterness and fishy odor reduce its acceptability to consumers. Here, CPI was first debittered using aminopeptidase from Streptomyces canus (ScAPase) and then deodorized through probiotic fermentation. This is the first reported demonstration of complete removal of the bitterness of CPI using ScAPase. Subsequently, Syn3 and Syn4, as aromatic CPI (ACPI), were prepared from debittered CPI (DCPI) via fermentation with Lactobacillus acidophilus and Bifidobacterium longum, respectively. These products, DCPI and ACPI, were characterized by the absence of bitterness and fishy odor, along with a strong aromatic scent and high overall acceptability. Additionally, these products exhibited improved physicochemical properties, including enhanced oil-holding capacity, emulsifying activity, and resistance to digestion, compared to untreated CPI. However, significant differences were observed in their radical scavenging activities. The highest scavenging activity was detected in Syn3 against DPPH˙ (63.5%) and ˙OH (79.2%), in DCPI against O2- (32.0%), and in post-digestion Syn4 against ABTS˙+ (95.2%). Furthermore, after digestion treatment, these products significantly promoted the proliferation of probiotics. Notably post-digestion Syn4 showed the most substantial proliferation effect on Lactobacillus reuteri, Lactobacillus rhamnosus, and Bifidobacterium breve compared to other post-digestion samples. These results indicate that the treated CPI has the potential for applications in health food products.

The BnaBPs gene regulates flowering time and leaf angle in Brassica napus.

The flowering time and plant architecture of Brassica napus were significantly associated with yield. In this study, we found that the BREVIPEDICELLUS/KNAT1(BP) gene regulated the flowering time and plant architecture of B. napus. However, the precise regulatory mechanism remains unclear. We cloned two homologous BP genes, BnaBPA03 and BnaBPC03, from B. napus Xiaoyun. The protein sequence analysis showed two proteins containing conserved domains KNOX I, KNOX II, ELK, and HOX of the KONX protein family. The CRISPR/Cas9 knockout lines exhibited early budding and flowering time, coupled with floral organ abscission earlier and a larger leaf angle. On the contrary, overexpression plants displayed a phenotype that was the inverse of these characteristics. Furthermore, we observed upregulation of gibberellin and ethylene biosynthesis genes, as well as floral integrator genes in knocked-out plants. The results revealed that BnaBPs play a role in flowering time, floral organ abscission, and leaf angle as well as germination processes mediated. Additionally, BnaBPs exerted an impact on the biosynthesis pathways of ethylene and GA.

α­Fetoprotein­positive hepatoid adenocarcinoma of the stomach and a new classification: A case report.

α-Fetoprotein (AFP)-producing gastric carcinoma (AFPGC) is a rare subtype of gastric cancer (GC) with controversial classification methods. Hepatoid adenocarcinoma of the stomach (HAS) is another rare subtype of GC. Its definition intersects with that of AFPGC, but it is much rarer. The present report describes the case of an elderly patient with GC and AFPGC and HAS features in a serum test and pathology, respectively, and proposes a new classification of GC subtypes based on histological and AFP-producing features. A 75-year-old woman presented with a history of polydipsia and polyuria for over a decade and dizziness for 1 day. Serum AFP levels gradually elevated from 183.70 to 397.70 ng/ml in 1 month after the patient's first clinic visit. Subsequent pathological findings from endoscopic biopsy samples confirmed a hepatoid focus with positive immunohistochemical staining for AFP. The patient underwent a laparoscopic-assisted radical total gastrectomy and esophagojejunal Roux-en-Y anastomosis, and the serum AFP levels decreased to the normal range after the surgery. The present case indicates the diagnostic value of both the serum AFP level and pathological examinations in the diagnosis of AFPGC and HAS, and also highlights the contemporary circumstances of the vague classification based on different criteria for these two subtypes. Furthermore, the present report proposes a new classification considering both histological and AFP-producing features (using both serum biomarkers and immunohistochemistry tests) to cover all cases encompassed by AFPGC and HAS under all definition methods. This new method would give more precise diagnoses and add value to the subsequent treatment decision-making.

Decoding the role of HIF-1α in immunoregulation in Litopenaeus vannamei under hypoxic stress.

Hypoxia poses a significant challenge to aquatic organisms, especially Litopenaeus vannamei (L. vannamei), which play a vital role in the global aquaculture industry. Hypoxia-inducible factor 1α (HIF-1α) is a pivotal regulator of the organism's adaptation to hypoxic conditions. To understand of how HIF-1α affects the immunity of L. vannamei under hypoxic conditions, we conducted a thorough study involving various approaches. These included observing tissue morphology, analyzing the expression of immune-related genes, assessing the activities of immune-related enzymes, and exploring immune-related pathways. Our study revealed that RNA interference (RNAi)-mediated knockdown of HIF-1α markedly reduced HIF-1α expression in the gill (75-95 %), whereas the reduction ranged from 2 to 43 % in the hepatopancreas. Knockdown of HIF-1α resulted in increased damage to both gill and hepatopancreatic tissues in hypoxic conditions. Additionally, immune-related genes, including Astakine (AST), Hemocyanin (HC), and Ferritin (FT), as well as immune-related enzymes such as Acid Phosphatase (ACP), Alkaline Phosphatase (AKP), and Phenoloxidase (PO), exhibited intricate regulatory patterns in response to hypoxia stress following the knockdown of HIF-1α. Transcriptome analysis revealed that HIF-1α knockdown significantly impacts multiple signaling pathways, including the JAK-STAT signaling pathway, Th17 cell differentiation pathways, PI3K-Akt signaling pathway, ErbB signaling pathway, MAPK signaling pathway, chemokine signaling pathway, ribosomal pathways, apoptosis, lysosomes and arachidonic acid metabolism. These alterations disrupt the organism's immune balance and interfere with normal metabolic processes, potentially leading to various immune-related diseases. We speculate that the weakened immune response resulting from HIF-1 inhibition is due to the reduced metabolic capacity, and the existence of a direct regulatory relationship between them requires further exploration. This study greatly advances our understanding of the vital role that HIF-1α plays in regulating immune responses in shrimp under hypoxic conditions, thereby deepening our comprehension of this critical biological mechanism.

Calcium regulates the physiological and molecular responses of Morus alba roots to cadmium stress.

Heavy metal cadmium (Cd) is toxic to organisms. Mulberry (Morus alba L.) is a fast-growing perennial that is also an economical Cd phytoremediation material with large biomass. However, the molecular mechanisms underlying its Cd tolerance remain unclear. Here, we reveal the physiological and molecular mechanisms underlying Cd toxicity under varying calcium (Ca) treatments. First, under low-Ca treatment (0.1 mM Ca), mulberry growth was severely inhibited and the root surface structure was damaged by Cd stress. Second, electrophysiological data demonstrated that 0.1 mM Ca induced an increased Cd2+ influx, leading to its accumulation in the entire root and root cell walls. Third, high-Ca treatment (10 mM Ca) largely alleviated growth inhibition, activated antioxidant enzymes, increased Ca content, decreased Cd2+ flux, and inhibited Cd uptake by roots. Finally, 0.1 mM Ca resulted in the activation of metal transporters and the disruption of Ca signaling-related gene expression, which facilitated Cd accumulation in the roots, aggravating oxidative stress. These adverse effects were reversed by treatment with 10 mM Ca. This study preliminarily revealed the mechanism by which varying Ca levels regulate Cd uptake and accumulation in mulberry roots, provided an insight into the interrelationships between Ca and Cd in the ecological and economic tree mulberry and offered a theoretical basis for Ca application in managing Cd pollution.

Prevalence and risk factors of symptomatic venous thromboembolism in distal femur fractures.

INTRODUCTION: There is a paucity of published research on symptomatic venous thromboembolism (sVTE) after distal femur fractures (DFFs). This study aimed to explore the prevalence and risk factors of sVTE in DFFs. METHODS: We identified a total of 131 patients who underwent DFF surgeries without routine pharmacological thromboprophylaxis between October 2007 and November 2016. Cases of sVTE included symptomatic pulmonary embolism (sPE) and symptomatic deep vein thrombosis (sDVT). Patients with sVTE were compared to those without, and differences in demographics and fracture-related characteristics were explored. Multivariate logistic regression was used to eliminate confounding factors. RESULTS: Of the 131 patients, 20 (15.3%) had sVTE, of whom 16 (12.2%) had sDVT and six (4.6%) had sPE (two patients had both sPE and sDVT). Notably, 17 (85.0%) sVTE patients were aged ≥60 years, while only 62 (55.9%) non-sVTE patients were aged ≥60 years (P = 0.014). Fourteen (82.4%) patients with sVTE had body mass index (BMI) ≥25 kg/m2, while 49 (53.3%) patients without sVTE had BMI ≥25 kg/m2 (P = 0.032). Multivariate logistic regression demonstrated that age ≥60 years (adjusted odds ratio [OR] 5.05; P = 0.040) and BMI ≥25 kg/m2 (adjusted OR 3.92; P = 0.045) were independently associated with a higher risk of sVTE after DFF. CONCLUSION: The prevalence of sVTE in DFFs is high at 15.3%. Advanced age (≥60 years) and being overweight (BMI ≥25 kg/m2) were two independent risk factors for sVTE in DFFs. The use of routine pharmacological thromboprophylaxis should be considered as a preventative measure.

Changes in activity and microbial community composition of Anammox-HAP granules during long-term preservation under different conditions.

Preservation of anammox granular sludge is important for anammox technology applications. Although previous studies have explored preservation methods, their long-term effects on microbial communities and functional genes remain underexplored. This study investigated the long-term preservation of anammox-hydroxyapatite (HAP) granules with storage durations of up to six years and examined the effects of different preservation methods. Results show that 4°C with 5 mM molybdate not only prevented the blackening of granules but also maintained a lower decay rate of specific anammox activity, preserving >50% after 6 months and 10% after 1 year. Functional gene analysis revealed an increase in sulfur-reducing genes such as dcyD and NADPH, particularly in samples without molybdate. These changes may result in hydrogen sulfide production, which contributes to sludge blackening. This study provides critical insights into optimizing the preservation conditions for anammox-HAP granules, facilitating rapid reactor start-up, and offering a viable solution for long-term storage.

Cystitis glandularis: MR imaging characteristics in 27 patients.

PURPOSE: To explore the diagnostic characteristics of cystitis glandularis (CG) using magnetic resonance imaging (MRI). MATERIALS AND METHODS: A retrospective study was conducted on pathologically confirmed patients who underwent bladder MRI examination between January 2019 and November 2023. Image analysis was jointly conducted, with emphasis on lesion location, morphology, size, signal intensity, and pattern of enhancement, by two genitourinary radiologists with 22 and 15 years of experience, respectively. RESULTS: A total of 27 patients with 27 lesions were included (median age 47 years, 24 males). The lesions were mostly located in the bladder trigone area (18/27). The lesions could be categorized as focal thickening (17/27), nodular (8/27), and diffuse thickening of the entire bladder (2/27) in morphological terms. On T2-weighted imaging (T2WI), 15 of 17 focal thickening lesions appeared as a slightly hyperintense thickened inner layer, with a higher signal in the center of the thickened inner layer, resembling a sandwich sign, and 6 of 8 nodular lesions were slightly hyperintense. On T1-weighted imaging (T1WI), 19 patients showed slight hypointensity. The lesions on DWI showed mainly high (5/27) and slightly high signal (21/27), with an average mean apparent diffusion coefficient (mADC) value of 2.171 ± 0.052 × 10-3mm2/s. Among the 23 patients who underwent dynamic contrast-enhanced (DCE) scanning, 18 lesions showed mild enhancement in the arterial phase (average 1.7 times comparing to unenhanced phase), and the degree of enhancement gradually increased in the venous and delayed phases (average 2.2 and 2.3 times compared to the unenhanced phase, respectively), showing a progressive enhancement pattern. CONCLUSION: On MRI, the majority of CG manifest as focal thickening or nodules in the bladder trigone area, showing slight hyperintensity on T2WI, slight hypointensity on T1WI, and a progressive enhancement pattern, without significant restriction on DWI. Focal thickening lesions may exhibit a special sandwich sign.

Cimicifuga heracleifolia kom. Attenuates ulcerative colitis through the PI3K/AKT/NF-κB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Cimicifuga heracleifolia Kom. (C. heracleifolia) has demonstrated efficacy in treating gastrointestinal disorders, including splenasthenic diarrhea. Ulcerative colitis (UC), a chronic inflammatory bowel disease, shares similarities with splenasthenic diarrhea. However, the pharmacological effects of C. heracleifolia on UC and the underlying mechanisms remain unexplored. AIM OF THE STUDY: The present study investigates the therapeutic potential and mechanisms of C. heracleifolia in UC. METHODS: Initially, network pharmacology analysis, encompassing ingredient screening, target prediction, protein-protein interaction (PPI) network analysis, and enrichment analysis, was employed to predict the mechanisms of C. heracleifolia. The findings were further validated using transcriptomics and functional assays in a dextran sulfate sodium (DSS)-induced UC model. Additionally, bioactive compounds were identified through surface plasmon resonance (SPR) analysis, molecular docking, and cell-based assays. RESULTS: A total of 52 ingredients of C. heracleifolia were screened, and 32 key targets were identified within a PPI network comprising 285 potential therapeutic targets. Enrichment analysis indicated that the anti-UC effects of C. heracleifolia are mediated through immune response modulation and the inhibition of inflammatory signaling pathways. In vivo experiments showed that C. heracleifolia mitigated histological damage in the colon, reduced the expression of phosphorylated Akt1, nuclear factor-kappa B (NF-κB) p65, and inhibitor of Kappa B kinase α/ß (IKKα/ß), suppressed the content of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), and enhanced the expression of tight junction proteins. Moreover, cimigenoside, caffeic acid, and methyl caffeate were identified as the bioactive constituents responsible for the UC treatment effects of C. heracleifolia. CONCLUSIONS: In summary, this study is the first to demonstrate that C. heracleifolia exerts therapeutic effects on UC by enhancing the intestinal mucosal barrier and inhibiting the phosphatidylinositol 3-kinase (PI3K)/AKT/NF-κB signaling pathway. These findings offer valuable insights into the clinical application of C. heracleifolia for UC management.

A new holistic perspective to assess the ecological risk of microplastics: A case study in Baiyangdian Basin, China.

By integrating probabilistic ecological risk assessment with the overall risk index method, which considers the multidimensional characteristics of the microplastome, the ecological risks of microplastic pollution were assessed more comprehensively. This study took the Baiyangdian Basin as an example to address the limitations of current risk assessment methods that rely on concentration data or the individual risk of microplastics. Using an exponential regression model, the acute and chronic ecological risk thresholds for the overall risk index method were determined to be 0.43 and 0.30, respectively. The acute and chronic ecological risks of the microplastome occupied 61 % and 79 % of the Baiyangdian Wetland and 0 % and 14 % of the Fu River, while the Xiaoyi River did not exhibit risk during the rainy season. Results indicated that intense human activities, poor hydrodynamics, low settling velocity and high levels of environmental chemical pollutants jointly contributed to the high risk of the microplastome in water bodies. Compared with the probabilistic ecological risk assessment method (risk characterization ratio), there was a significant difference in the area of acute and chronic ecological risks caused by the microplastome in the Baiyangdian Basin when using the overall risk index method. This proved that considering only concentration cannot truly reflect the toxicity of microplastics to aquatic organisms.

Multiomic single cell sequencing identifies stemlike nature of mixed phenotype acute leukemia.

Despite recent work linking mixed phenotype acute leukemia (MPAL) to certain genetic lesions, specific driver mutations remain undefined for a significant proportion of patients and no genetic subtype is predictive of clinical outcomes. Moreover, therapeutic strategy for MPAL remains unclear, and prognosis is overall poor. We performed multiomic single cell profiling of 14 newly diagnosed adult MPAL patients to characterize the inter- and intra-tumoral transcriptional, immunophenotypic, and genetic landscapes of MPAL. We show that neither genetic profile nor transcriptome reliably correlate with specific MPAL immunophenotypes. Despite this, we find that MPAL blasts express a shared stem cell-like transcriptional profile indicative of high differentiation potential. Patients with the highest differentiation potential demonstrate inferior survival in our dataset. A gene set score, MPAL95, derived from genes highly enriched in the most stem-like MPAL cells, is applicable to bulk RNA sequencing data and is predictive of survival in an independent patient cohort, suggesting a potential strategy for clinical risk stratification.

Genome-Wide Identification of Pentatricopeptide Repeat (PPR) Gene Family and Multi-Omics Analysis Provide New Insights Into the Albinism Mechanism of Kandelia obovata Propagule Leaves.

Pentatricopeptide repeat (PPR) gene family constitutes one of the largest gene families in plants, which mainly participate in RNA editing and RNA splicing of organellar RNAs, thereby affecting the organellar development. Recently, some evidence elucidated the important roles of PPR proteins in the albino process of plant leaves. However, the functions of PPR genes in the woody mangrove species have not been investigated. In this study, using a typical true mangrove Kandelia obovata, we systematically identified 298 PPR genes and characterized their general features and physicochemical properties, including evolutionary relationships, the subcellular localization, PPR motif type, the number of introns and PPR motifs, and isoelectric point, and so forth. Furthermore, we combined genome-wide association studies (GWAS) and transcriptome analysis to identify the genetic architecture and potential PPR genes associated with propagule leaves colour variations of K. obovata. As a result, we prioritized 16 PPR genes related to the albino phenotype using different strategies, including differentially expressed genes analysis and genetic diversity analysis. Further analysis discovered two genes of interest, namely Maker00002998 (PLS-type) and Maker00003187 (P-type), which were differentially expressed genes and causal genes detected by GWAS analysis. Moreover, we successfully predicted downstream target chloroplast genes (rps14, rpoC1 and rpoC2) bound by Maker00002998 PPR proteins. The experimental verification of RNA editing sites of rps14, rpoC1, and rpoC2 in our previous study and the verification of interaction between Maker00002998 and rps14 transcript using in vitro RNA pull-down assays revealed that Maker00002998 PPR protein might be involved in the post-transcriptional process of chloroplast genes. Our result provides new insights into the roles of PPR genes in the albinism mechanism of K. obovata propagule leaves.

Self-Assembled PEGylated Nanocubes Based on Hydrophobic Camptothecin and Doxorubicin for Combinational Therapy of Colorectal Cancer.

Nanoassemblies based on drug conjugates with high drug loading efficiency and stability have been regarded as promising candidates for the next generation of drug formulations. However, they are mostly amphiphilic. Here, a dual-hydrophobic drug conjugate-based nanoassembly has been created for enhanced synergistic antiproliferation against colorectal cancer cells. Camptothecin (CPT) and doxorubicin (DOX) were chosen as the hydrophobic drugs and covalently linked with a disulfide bond (-ss-). The synthesized CPT-ss-DOX can self-assemble into nanocubes (NCs) in an aqueous solution with the assistance of a small amount of polyethylene glycol (PEG), named PEGylated CPT-ss-DOX NCs. The PEGylated CPT-ss-DOX NCs were approximately 111.8 nm, possessing a crystal structure and a very low critical aggregation concentration (8.36 µg·mL-1). The self-assembly mechanism was studied using molecular docking and molecular dynamic simulation methods. The NCs demonstrated excellent storage stability and improved water solubility of CPT and DOX. These NCs could be taken up by cancer cells and gradually release the drugs. In addition, they had higher toxicity to cancer cells than a mixture of CPT and DOX, while they displayed reduced toxicity to normal cells. Due to assembly and PEG modification, the NCs improved drug retention time and enhanced accumulation at the tumor site. More importantly, they significantly inhibited colorectal tumor growth (58.37%) in vivo, superior to the CPT+DOX mix (42.63%). Moreover, the NCs reduced the cardiac toxicity of free drugs. Therefore, the prepared PEGylated CPT-ss-DOX NCs hold great potential for clinical transformation and provide a novel method for the self-delivery of hydrophobic molecules in cancer therapy.

Liquid-liquid phase separation-related genes associated with prognosis, tumor microenvironment characteristics, and tumor cell features in bladder cancer.

OBJECTIVE: This study aimed to explore the Liquid-liquid phase separation (LLPS)-related genes associated with the prognosis of bladder cancer (BCa) and assess the potential application of LLPS-related prognostic signature for predicting prognosis in BCa patients. METHODS: Clinical information and transcriptome data of BCa patients were extracted from the Cancer Genome Atlas-BLCA (TCGA-BLCA) database and the GSE13507 database. Furthermore, 108 BCa patients who received treatment at our institution were subjected to a retrospective analysis. The least absolute shrinkage and selection operator (LASSO) analysis was performed to develop an LLPS-related prognostic signature for BCa. The CCK8, wound healing and Transwell assays were performed. RESULTS: Based on 62 differentially expressed LLPS-related genes (DELRGs), three DELRGs were screened by LASSO analysis including kallikrein-related peptidase 5 (KLK5), monoacylglycerol O-acyltransferase 2 (MOGAT2) and S100 calcium-binding protein A7 (S100A7). Based on three DELRGs, a novel LLPS-related prognostic signature was constructed for individualized prognosis assessment. Kaplan-Meier curve analyses showed that LLPS-related prognostic signature was significantly correlated with overall survival (OS) of BCa. ROC analyses demonstrated the LLPS-related prognostic signature performed well in predicting the prognosis of BCa patients in the training group (the area under the curve (AUC) = 0.733), which was externally verified in the validation cohort 1 (AUC = 0.794) and validation cohort 2 (AUC = 0.766). Further experiments demonstrated that inhibiting KLK5 could affect the proliferation, migration, and invasion of BCa cells. CONCLUSIONS: In this study, a novel LLPS-related prognostic signature was successfully developed and validated, demonstrating strong performance in predicting the prognosis of BCa patients.

The influence of COVID-19 on short-term mortality in acute ischemic stroke: A systematic review and meta-analysis.

OBJECTIVE: To evaluate the differences in short-term mortality risk between acute ischemic stroke (AIS) patients with and without SARS-CoV-2 infection. METHODS: PubMed, EMBASE, Scopus, and Cochrane Databases were systematically searched from December 1, 2019 to May 20, 2022 using the keywords coronavirus disease 2019 (COVID-19), COVID-19, SARS-CoV-2, and ischemic stroke. A random-effects model was estimated, and subgroup analysis and meta-regressions were performed. The quality of eligible studies was assessed using the Newcastle-Ottawa Scale. RESULTS: A total of 26 eligible studies with 307,800 patients were included in this meta-analysis. The overall results show that in-hospital and 90-day mortality was 3.31-fold higher in AIS with SARS-CoV-2 patients compared with those without SARS-CoV-2. When matched for age and National Institutes of Health Stroke Scale score at admission, the risk ratio of in-hospital mortality from AIS among patients with SARS-CoV-2 versus without decreased to 2.83. Reperfusion therapy and endovascular thrombectomy may further reduce the risk of death in patients to some extent but do not increase the incidence of symptomatic intracerebral hemorrhage. Meta-regression showed that in-hospital mortality decreased with increasing National Institutes of Health Stroke Scale score in AIS with SARS-CoV-2 compared to those without SARS-CoV-2 and that the difference in mortality risk between the 2 was independent of age and sex. CONCLUSIONS: The results of this study suggest that AIS patients with SARS-CoV-2 have higher short-term mortality compared to AIS patients without SARS-CoV-2, and reperfusion and endovascular thrombectomy therapy may reduce the risk of short-term mortality to some extent. The differences in in-hospital mortality risk were similar across ages and sexes. Focused attention is therefore needed on AIS patients with SARS-CoV-2 to control mortality.

Crystal structure of the Rib domain of the cell-wall-anchored surface protein from Limosilactobacillus reuteri.

The immunoglobulin (Ig)-like domain is found in a broad range of proteins with diverse functional roles. While an essential ß-sandwich fold is maintained, considerable structural variations exist and are critical for functional diversity. The Rib-domain family, primarily found as tandem-repeat modules in the surface proteins of Gram-positive bacteria, represents another significant structural variant of the Ig-like fold. However, limited structural and functional exploration of this family has been conducted, which significantly restricts the understanding of its evolution and significance within the Ig superclass. In this work, a high-resolution crystal structure of a Rib domain derived from the probiotic bacterium Limosilactobacillus reuteri is presented. This protein, while sharing significant structural similarity with homologous domains from other bacteria, exhibits a significantly increased thermal resistance. The potential structural features contributing to this stability are discussed. Moreover, the presence of two copper-binding sites, with one positioned on the interface, suggests potential functional roles that warrant further investigation.

Chronobiological perspectives: Association between meal timing and sleep quality.

BACKGROUND: Meal timing has been associated with metabolism and cardiovascular diseases; however, the relationship between meal timing and sleep quality remains inconclusive. OBJECTIVE: This study aims to investigate the relationship between meal timing and sleep quality from a chronobiological perspective. METHODS: This study utilized data from the NHANES for the years 2005-2008, including a cohort of 7,023 participants after applying exclusion criteria. Sleep quality was assessed using the Pittsburgh Sleep Quality Index (PSQI). Meal timing was analyzed based on two 24-hour dietary recalls from each individual, considering the timing of the initial and final meals, meal duration, and frequency of meal occasions. Multiple linear regression models and hierarchical analyses were employed to examine the relationship between meal timing and PSQI scores, adjusting for various demographic and habitat covariates. RESULTS: Statistical analysis revealed a positive correlation between delayed meal timings, increased meal occasions, and elevated PSQI scores, indicating that later meal timing are intricately linked with diminished sleep quality. Both later meal timings and more frequent meal occasions were significantly associated with poorer sleep quality. Compared to the first tertile, the ß (95%CI) values of the third tertile were 0.545 (0.226, 0.864) for first meal timing, 0.586 (0.277, 0.896) for midpoint meal timing, 0.385 (0.090, 0.680) for last meal timing, and 0.332 (0.021, 0.642) for meal occasions in the adjusted models. CONCLUSION: These findings suggest that late initial, midpoint, and final meal timing, as well as more frequent meal occasions, are chrono-nutrition patterns associated with poor sleep quality.