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As the global demographic shifts toward an aging population, understanding the efficiency of protein utilization in older adults becomes crucial. Our study explores the intricate relationship between protein intake and aging, with a focus on precision nutrition for older people. Through a meta-analysis, we confirm a decline in protein-utilization capacity in older individuals and examine the different contributions of plant and animal protein. In experiments involving mice of different ages, older mice exhibited decreases in the biological utilization of four proteins (casein, beef protein, soy protein, and gluten), particularly casein. In subsequent research, casein was studied as a key protein. A decline in gastric digestion function was observed through peptidomics and the examination of pepsin levels using casein. Nevertheless, this decline did not significantly affect the overall protein digestion during the aging process. The combined application of targeted amino acid metabolomics identified abnormal absorption of amino acids as the underlying cause of decreased protein utilization during aging, particularly emphasizing a reduction in branched-chain amino acids (BCAAs) in older mice. Delving deeper into the proteomics of the intestinal protein digestion and absorption pathway, a reduction of over 60% in large neutral amino acid transporter 2 (LAT2) protein expression was observed in both older humans and aged mice. The reduction in LAT2 protein was found to be a key factor influencing the diminished BCAA availability. Overall, our study establishes the significance of amino acid absorption through LAT2 in protein utilization during aging and offers a new theoretical foundation for improving protein utilization in the older adults.

Abstract Anthocidaris crassispina is a very popular edible sea urchin distributed along the coast of the South China Sea. In this study, we performed whole-genome sequencing and generated a chromosome-level assembly of this species. The total length of the genomic contig sequence was 891.02 Mb, and contig N50 was 808.15 kb when Hifiasm was used for assembly. The Hi-C library was constructed and sequenced, yielding approximately 68.61 Gb of data. After Hi-C assembly, approximately 886.72 Mb of sequence was able to be mapped onto 21 chromosomes, accounting for 99.52% of the total genome length. Among the sequences located on the chromosomes, those for which the order and direction could be determined accounted for approximately 826.82 Mb, or 93.24% of the total length. These results provide valuable resources for further study of A. crassispina at the genetic level.

Abstract Developing effective electrocatalysts for the nitrate reduction reaction (NO3RR) is a promising alternative to conventional industrial ammonia (NH3) synthesis. Herein, starting from a flexible laser‐induced graphene (LIG) film with hierarchical and interconnected macroporous architecture, a binder‐free and free‐standing Ru‐modified LIG electrode (Ru‐LIG) is fabricated for electrocatalytic NO3RR via a facile electrodeposition method. The relationship between the laser‐scribing parameters and the NO3RR performance of Ru‐LIG electrodes is studied in‐depth. At −0.59 VRHE, the Ru‐LIG electrode exhibited the optimal and stable NO3RR performance (NH3 yield rate of 655.9 µg cm−2 h−1 with NH3 Faradaic efficiency of up to 93.7%) under a laser defocus setting of +2 mm and an applied laser power of 4.8 W, outperforming most of the reported NO3RR electrodes operated under similar conditions. The optimized laser‐scribing parameters promoted the surface properties of LIG with increased graphitization degree and decreased charge‐transfer resistance, leading to synergistically improved Ru electrodeposition with more exposed NO3RR active sites. This work not only provides a new insight to enhance the electrocatalytic NO3RR performance of LIG‐based electrodes via the coordination with metal electrocatalysts as well as identification of the critical laser‐scribing parameters but also will inspire the rational design of future advanced laser‐induced electrocatalysts for NO3RR.

The objective of this study was to assess the impact of Lactobacillus plantarum, Bacillus subtilis, and a lignocellulolytic enzyme system on the nutritional value, fermentation profiles, rumen digestion, and bacterial community of fresh waxy corn stalk silage. Fresh waxy corn stalks harvested after 90 days of growth were treated with no additives (CON), compound multiple lactobacilli (ML, comprising Lactobacillus plantarum at 1.0 × 105 cfu/g fresh weight and Bacillus subtilis at 1.0 × 105 cfu/g fresh weight), a lignocellulolytic enzyme system (LE, at 500 g/t), and a combination of the lignocellulolytic enzyme system and multiple lactobacilli (MLLE). Throughout the fermentation process, the contents of dry matter (DM) and water-soluble carbohydrates (WSC), as well as the counts of lactic acid bacteria, yeast, and mold, continuously decreased with the extension of the fermentation time. In contrast, the concentrations of acetic acid, propionic acid, and ammonia N progressively increased over time. Notably, the pH initially decreased and then increased as fermentation progressed, while the lactic acid concentration first increased and then decreased with the extension of the fermentation time. After 45 days of fermentation, the DM contents of LE and MLLE were significantly lower than that of CON, while the WSC content of MLLE was significantly higher than that of CON. LE most effectively increased the ED of DM and neutral detergent fiber (NDF). Compared to ML and LE, MLLE showed a greater increase in the “a” value of DM and NDF. The DM content in LE and MLLE was lower than in CON, with a higher WSC content. Both ML and MLLE produced more lactic acid than CON and LE, resulting in a lower pH in these groups. Additionally, the acetic acid content and ammonia N content were lower in ML and MLLE compared to CON and LE. The counts of lactic acid bacteria, yeast, and mold declined over the fermentation period, dropping below detectable levels on the seventh and fifth days, respectively. Relative to CON, the three additives reduced the relative abundance of Weissella and Klebsiella and increased that of Lactobacillus. In conclusion, LE improved the nutritional value and rumen digestion of the silage, ML enhanced its fermentation profiles and bacterial community, and MLLE significantly improved the nutritional value, rumen digestion characteristics, fermentation profiles, and bacterial community of the fresh waxy corn stalk silage.

Abstract The clinical applications of the association of cortical thickness and white matter fiber with freezing of gait (FoG) are limited in patients with Parkinson’s disease (PD). In this retrospective study, using white matter fiber from diffusion-weighted imaging and cortical thickness from structural-weighted imaging of magnetic resonance imaging, we investigated whether a machine learning-based model can help assess the risk of FoG at the individual level in patients with PD. Data from the Parkinson’s Disease Progression Marker Initiative database were used as the discovery cohort, whereas those from the Fujian Medical University Union Hospital Parkinson’s Disease database were used as the external validation cohort. Clinical variables, white matter fiber, and cortical thickness were selected by random forest regression. The selected features were used to train the support vector machine(SVM) learning models. The median area under the receiver operating characteristic curve (AUC) was calculated. Model performance was validated using the external validation cohort. In the discovery cohort, 25 patients with PD were defined as FoG converters (15 men, mean age 62.1 years), whereas 60 were defined as FoG nonconverters (38 men, mean age 58.5 years). In the external validation cohort, 18 patients with PD were defined as FoG converters (8 men, mean age 66.9 years), whereas 37 were defined as FoG nonconverters (21 men, mean age 65.1 years). In the discovery cohort, the model trained with clinical variables, cortical thickness, and white matter fiber exhibited better performance (AUC, 0.67–0.88). More importantly, SVM-radial kernel models trained using random over-sampling examples, incorporating white matter fiber, cortical thickness, and clinical variables exhibited better performance (AUC, 0.88). This model trained using the above mentioned features was successfully validated in an external validation cohort (AUC, 0.91). Furthermore, the following minimal feature sets that were used: fractional anisotropy value and mean diffusivity value for right thalamic radiation, age at baseline, and cortical thickness for left precentral gyrus and right dorsal posterior cingulate gyrus. Therefore, machine learning-based models using white matter fiber and cortical thickness can help predict the risk of FoG conversion at the individual level in patients with PD, with improved performance when combined with clinical variables.

The molecular landscape of microphthalmia transcription factor family translocation renal cell carcinoma tumours remain to be characterised. Here, the authors perform proteogenomic analysis and reveal dysregulation of DNA repair, mTOR signalling and metabolic processes.

Background: The genomic background affects the occurrence and metastasis of cancers, including clear cell renal cell carcinoma (ccRCC). However, reports focusing on the prognostic mutational signature of Chinese ccRCC are lacking. Methods: Overall, 929 patients, including a training cohort with Chinese patients (n = 201), a testing cohort with Caucasian patients (n = 274), and a validation cohort (n = 454) were analyzed for the genomic landscape of ccRCC. Then, machine-learning algorithms were used to identify and evaluate the genomic mutational signature (GMS) in ccRCC. Analyses for prognosis, immune microenvironment, association with independent clinicopathological features, and predictive responses for immune checkpoint therapies (ICTs) were performed. Results: The DNA variation data of 929 patients with ccRCC suggested markedly differential genomic mutational frequency of the most frequent genes, such as VHL, PBRM1, BAP1, SETD2, and KDM5C between the Chinese and Caucasian populations. PBRM1 showed significant co-occurrence with VHL and SETD2. We then successfully identified a seven-gene mutational signature (GMSMut) that included mutations in FBN1, SHPRH, CELSR1, COL6A6, DST, ABCA13, and BAP1. The GMSMut significantly predicted progressive progression (P < 0.0001, HR = 2.81) and poor prognosis (P < 0.0001, HR = 3.89) in the Chinese training cohort. Moreover, ccRCC patients with the GMSMut had poor survival rates in the testing cohort (P = 0.020) and poor outcomes were predicted for those treated with ICTs in the validation cohort (P = 0.036). Interestingly, a favorable clinical response to ICTs, elevated expression of immune checkpoints, and increased abundance of tumor-infiltrated lymphocytes, specifically CD8+ T cells, Tregs, and macrophages, were observed in the GMSMut cluster. Conclusions: This study described the pro-tumorigenic GMSMut cluster that improved the prognostic accuracy in Chinese patients with ccRCC. Our discovery of the novel independent prognostic signature highlights the relationship between tumor phenotype and genomic mutational characteristics of ccRCC.

Abstract Clear cell renal cell carcinoma (ccRCC) is one of the most common malignant genitourinary cancers with high recurrence risk worldwide. Recently, multi-omics data facilitate obtaining a molecular landscape of tumor development, and were implemented to affect pathogenesis, phenotype, and prognosis of ccRCC. In this study, after screening for differential expressed microRNAs based on multiply datasets, we tested expression levels and prognostic value of miR-187-3p in ccRCC samples, and transfected miR-187-3p mimics or negative controls into ccRCC cells. Up-regulation of miR-187-3p restrains proliferation, migration and promotes apoptosis ability in human ccRCC A498 and 786O cells. In addition, Luciferase reporter assay revealed that miR-187-3p directly targets LRFN1-3’-UTR and negatively modulates LRFN1 expression. LRFN1 rescues proliferation and invasion capacities after miR-187-3p mimic transfection in vitro and in subcutaneous xenograft models. We further performed deep-sequencing technology and bioinformatics analyses to evaluate the biological functions and potential clinical implications of LRFN1 expression in ccRCC. Interestingly, LRFN1 could serve as an independent and potential biomarker for prognosis in over 1000 patients with ccRCC from multiply independent cohorts. Besides, the up-regulated LRFN1 expression prominently promoted intra-tumoral heterogeneity and immune-infiltrating microenvironment, represented by elevated M2 macrophage infiltration, CD8+ T cells activity and PD-L1 expression. In conclusion, this study revealed the tumor-specific and immunological role of miR-187-3p/LRFN1 axis in the progression and reshaping of tumor immune microenvironment of ccRCC.

Abstract Catalytic reactions are surface‐sensitive processes. Fabrication of homogeneous metastable metals can be used to promote phase‐dependent catalytic performance; however, this has been a challenging task. Herein, homogeneous metastable hexagonal close‐packed (hcp) Ir is epitaxially grown onto metastable phase hcp Ni, as demonstrated using spherical aberration electron microscopy. The as‐fabricated metastable hcp Ir exhibits high intrinsic activity for the alkaline hydrogen evolution reaction (HER). In particular, metastable hcp Ir delivers a low overpotential of 17 mV at 10 mA cm−2 and presents a high specific activity of 8.55 mA cm−2 and a high turnover frequency of 38.26 s−1 at −0.07 V versus the reversible hydrogen electrode. Owing to its epitaxially grown structure, metastable hcp Ir is highly stable. Theoretical calculations reveal that metastable hcp Ir promotes H2O adsorption and fast H2O dissociation, which contributes to its remarkable HER activity. Findings can elucidate the crystal phase‐controlled synthesis of advanced noble metal nanomaterials for the fundamental catalytic applications.

BackgroundIncreasing evidence indicates that L-dopa decarboxylase (DDC), which mediates aberrant amino acid metabolism, is significantly associated with tumor progression. However, the impacts of DDC are not elucidated clearly in clear cell renal cell carcinoma (ccRCC). This study aimed to evaluate DDC prognostic value and potential mechanisms for ccRCC patients.MethodsTranscriptomic and proteomic expressions of and clinical data including 532 patients with ccRCC (The Cancer Genome Atlas RNA-seq data), 226 ccRCC samples (Gene Expression Omnibus), 101 ccRCC patients from the E-MTAB-1980 cohort, and 232 patients with ccRCC with proteogenomic data (Fudan University Shanghai Cancer Center) were downloaded and analyzed to investigate the prognostic implications of DDC expression. Cox regression analyses were implemented to explore the effect of DDC expression on the prognosis of pan-cancer. The "limma" package identified the differentially expressed genes (DEGs) between high DDC subgroups and low DDC groups. Functional enrichments were performed based DEGs between DDC subgroups. The differences of immune cell infiltrations and immune checkpoint genes between DDC subgroups were analyzed to identify potential influence on immune microenvironment.ResultsWe found significantly decreased DDC expression in ccRCC tissues compared with normal tissues from multiple independent cohorts based on multi-omics data. We also found that DDC expression was correlated with tumor grades and stages.The following findings revealed that lower DDC expression levels significantly correlated with shorter overall survival (P

Background: Zein is the major protein in corn and the main byproduct of the starch industry and, therefore, crucial for global agricultural commodities. Its unique fibre-forming properties, which underline its potential as a biodegradable material, have drawn intensified interest from the public, scientists, and industries because of growing concern about environmental risks. As zein electrospun nanofibre has been studied in different research fields, the recent advances in zein electrospun nanofibres in the therapeutic field need re-examination. Scope and approach: This review provides an overview of various methods of modifications (e.g., crosslinking, deamidation and glycosylation on the fibre-forming properties of zein and different electrospun setups (e.g., emulsion and multi-layer electrospinning) in the last decade and reports current applications in the therapeutic field such as controlled release, wound healing, tissue engineering and biosensor. It also discussed the mechanisms of modifications such as electrostatic interactions, hydrophobic interactions, and van der Waals forces. Key findings and conclusions: The biocompatibility, fibre-forming and encapsulation properties of zein make it a potential replacement for traditional synthetic materials. Since specific properties of zein-based materials are acquirable via different modifications and processing, developing an insight into the relationship between functions and microstructure and conformations of the modified zein-based nanofibres along with different electrospun processes could contribute to high-quality zein-based fibrous products. It also discussed the potential limitations and challenges of zein-based nanofibres.

BackgroundRenal cancer is one of the most lethal cancers because of its atypical symptoms and metastatic potential. The metabolism of amino acids and their derivatives is essential for cancer cell survival and proliferation. Thus, the construction of the amino acid metabolism-related risk signature might enhance the accuracy of the prognostic model and shed light on the treatments of renal cancers.MethodsRNA expression and clinical data were downloaded from Santa Cruz (UCSC) Xena, GEO, and ArrayExpress databases. The “DESeq2” package identified the differentially expressed genes. Univariate COX analysis selected prognostic genes related to the metabolism of amino acids. Patients were divided into two clusters using the “ConsensusClusterPlus” package, and the CIBERSORT, ESTIMATE methods were explored to assess the immune infiltrations. The LASSO regression analysis constructed a risk model which was evaluated the prediction accuracy in two independent cohorts. The genomic alterations and drug sensitivity of 18-LASSO-genes were assessed. The differentially expressed genes between two clusters were used to perform functional enrichment analysis and weighted gene co-expression network analysis (WGCNA). Furthermore, external validation of TMEM72 expression was conducted in the FUSCC cohort containing 33 ccRCC patients.ResultsThe amino acid metabolism-related genes had significant correlations with prognosis. The patients in Cluster A demonstrated better survival, lower Treg cell proportion, higher ESTIMATE scores, and higher cuproptosis-related gene expressions. Amino acid metabolism-related genes with prognostic values were used to construct a risk model and patients in the low risk group were associated with improved outcomes. The Area Under Curve of the risk model was 0.801, 0.777, and 0.767 at the first, second, and third year respectively. The external validation cohort confirmed the stable prognostic value of the risk model. WGCNA identified four gene modules correlated with immune cell infiltrations and cuproptosis. We found that TMEM72 was downregulated in tumors by using TCGA, GEO datasets (p

In order to solve the increasingly serious environmental problems caused by plastic-based packaging, carrageenan-based films are drawing much attentions in food packaging applications, due to low cost, biodegradability, compatibility, and film-forming property. The purpose of this article is to present a comprehensive review of recent developments in carrageenan-based films, including fabrication strategies, physical and chemical properties and novel food packaging applications. Carrageenan can be extracted from red algae mainly by hydrolysis, ultrasonic-assisted and microwave-assisted extraction, and the combination of multiple extraction methods will be future trends in carrageenan extraction methods. Carrageenan can form homogeneous film-forming solutions and fabricate films mainly by direct coating, solvent casting and electrospinning, and mechanism of film formation was discussed in detail. Due to the inherent limitations of the pure carrageenan film, physical and chemical properties of carrageenan films were enhanced by incorporation with other compounds. Therefore, carrageenan-based films can be widely used for extending the shelf life of food and monitoring the food freshness by inhibiting microbial growth, reducing moisture loss and the respiration, etc. This article will provide useful guidelines for further research on carrageenan-based films.

Sarcomatoid differentiation is a highly aggressive pathological characteristic of renal cell carcinoma (RCC) and is characterized by susceptibility to progression and extremely poor prognosis. In this study, we included all genomic alteration events that led to a loss of protein function of MTAP and CDKN2A, and enrolled 5,307 RCC patients with genomic sequencing data from Western and Chinese cohorts. Notably, MTAP/CDKN2AMUT occurred in the Chinese population ~2 times more frequently than in the Western cohort and showed significant co-mutation trends. We found significantly higher proportions of sarcomatoid-positive patients with MTAPMUT or CDKN2AMUT compared with MTAP/CDKN2A wild-type (WT) patients (P

The complexation of protein and polysaccharide has shown considerable potential for the encapsulation of functional food components. In this work, propylene glycol alginate (PGA) molecules with different molecular weights (100, 500, and 2,000 kDa) were prepared through H2O2 oxidation, which were further combined with β-lactoglobulin nanoparticles (β-lgNPs) to form PGA-β-lgNPs complexes for the delivery of curcumin (Cur). Results showed that the depolymerization of PGA molecule was resulted from the breakage of glycosidic bonds in the main chain, and the depolymerization rate of PGA molecule depended on the reaction time, temperature, solution pH and H2O2 concentration. As the increasing molecular weight of PGA, the particle size, zeta-potential and turbidity of the complexes were obviously increased. The formation of PGA/β-lgNPs complexes was mainly driven by non-covalent interaction, including electrostatic gravitational interaction, hydrogen bonding and hydrophobic effect. Interestingly, the difference in the molecular weight of PGA also led to significantly differences in the micro-morphology of the complexes, as PGA with a high molecular weight (2,000 kDa) generated the formation of a “fruit-tree” shaped structure, whereas PGA with relatively low molecular weight (100 and 500 kDa) led to spherical particles with a “core-shell” structure. In addition, the incorporation of PGA molecules into β-lgNPs dispersion also contributed to the improvement in the encapsulation efficiency of Cur as well as physicochemical stability of β-lgNPs, and PGA with a higher molecular weight was confirmed with a better effect. Findings in the current work may help to further understand the effect of molecular weight of polysaccharide on the physical and structural properties as well as effectiveness as delivery systems of polysaccharide-protein complexes, providing for the possibility for the design and development of more efficient carriers for bioactive compounds in food system.

Bladder cancer is one of the most common genitourinary malignant cancers worldwide. Cell death processes, including apoptosis, ferroptosis, and necrosis, provide novel clinical and immunological insights promoting the management of precision medicine. Therefore, this study aimed to evaluate the transcriptomic profile of signatures in cell death pathways with significant prognostic implications in patients with bladder cancer from multiple independent cohorts (n = 1999). First, genes involved in apoptosis (n = 19), ferroptosis (n = 31), and necrosis (n = 6) were analyzed to evaluate the prognostic implications in bladder cancer. Significant genes were included to establish the cell-death index (CDI) of 36 genes that distinguished patients according to high and low risks. Survival analysis using the Kaplan-Meier curves clustered patients based on overall survival (18.8 vs. 96.7 months; hazard model [HR] = 3.12, P

To understand the unprecedented difference between 6-endo and 5-exo selectivity in hypervalent iodine (III) promoted fluorocyclization of unsaturated carboxylic acids or alcohols by difluoroiodotoluene, density functional theory (DFT) studies have been performed to systematically compare both the previous proposed “fluorination first and cyclization later” mechanism and the alternative “cyclization first and fluorination later” mechanism. Our results revealed that the selectivity is mechanism-dependent. The unsaturated alcohol prefers the fluorination first and the 6-endo-tet cyclization later pathway, leading to the experimentally observed 6-endo ether product. In contrast, the unsaturated carboxylic acid plausibly undergoes the 5-exo-trig cyclization first and the fluorination later to the experimentally observed 5-exo lactone product. The pKa property of the functional group of the substrate is found to play a key role in determining the reaction mechanism. The provided insights into the mechanism-dependent selectivity should help advance the development of fluorocyclization reactions with hypervalent iodine reagents.

The emulsion gels have attracted extensive interests due to their unique physical characters, remarkable stability, and control release properties of flavor and functional components compared to emulsions in liquid. In the current work, whey protein isolate (WPI)/κ-carrageenan (κ-CG) composite emulsion gels were fabricated based on the ultra-high-pressure (UHP) technology, in replacement of the traditional thermal, acid, or enzyme processing. Uniform composite emulsion gels could be fabricated by UHP above 400 MPa with minimum WPI and κ-CG concentrations of 8.0 and 1.0 wt%, respectively. The formation of UHP-induced emulsion gels is mostly attributed to the hydrophobic interaction and hydrogen bonding. The emulsion gels with different textures, rheology properties, and microstructures could be fabricated through adjusting the formulations (WPI concentration, κ-CG concentration, and oil phase fraction) as well as processing under different conditions (pressure and time). Afterward, curcumin-loaded emulsion gels were fabricated and subjected to an in vitro simulated gastrointestinal digestion in order to investigate the gastrointestinal fate of curcumin. In vitro simulated digestion results demonstrated that the UHP treatment significantly retarded the release of curcumin but had little impact on the bioaccessibility of curcumin. The results in this work provide useful information for the construction of emulsion gels through a non-thermal process, which showed great potential for the delivery of heat-sensitive bioactive components.

Background: The tumor microenvironment affects the occurrence and development of cancers, including clear cell renal cell carcinoma (ccRCC). However, how the immune contexture interacts with the cancer phenotype remains unclear.Methods: We identified and evaluated immunophenotyping clusters in ccRCC using machine-learning algorithms. Analyses for functional enrichment, DNA variation, immune cell distribution, association with independent clinicopathological features, and predictive responses for immune checkpoint therapies were performed and validated.Results: Three immunophenotyping clusters with gradual levels of immune infiltration were identified. The intermediate and high immune infiltration clusters (Clusters B and C) were associated with a worse prognosis for ccRCC patients. Tumors in the immune-hot Clusters B and C showed pro-tumorigenic immune infiltration, and these patients showed significantly worse survival compared with patients in the immune-cold Cluster A in the training and testing cohorts (n = 422). In addition to distinct immune cell infiltrations of immunophenotyping, we detected significant differences in DNA variation among clusters, suggesting a high degree of genetic heterogeneity. Furthermore, expressions of multiple immune checkpoint molecules were significantly increased. Clusters B and C predicted favorable outcomes in 64 ccRCC patients receiving immune checkpoint therapies from the FUSCC cohort. In 360 ccRCC patients from the FUSCC validation cohort, Clusters B and C significantly predicted worse prognosis compared with Cluster A. After immunophenotyping of ccRCC was confirmed, significantly increased tertiary lymphatic structures, aggressive phenotype, elevated glycolysis and PD-L1 expression, higher abundance of CD8+ T cells, and TCRn cell infiltration were found in the immune-hot Clusters B and C.Conclusion: This study described immunophenotyping clusters that improved the prognostic accuracy of the immune contexture in the ccRCC microenvironment. Our discovery of the novel independent prognostic indicators in ccRCC highlights the relationship between tumor phenotype and immune microenvironment.

The tumor microenvironment (TME) affects the biologic malignancy of clear cell renal cell carcinoma (ccRCC). The influence of the 5-methylcytosine (m5C) epigenetic modification on the TME is unknown. We comprehensively assessed m5C modification patterns of 860 ccRCC samples (training, testing, and real-world validation cohorts) based on 17 m5C regulators and systematically integrated the modification patterns with TME cell-infiltrating characterizations. Our results identified distinct m5C modification clusters with gradual levels of immune cell infiltration. The distinct m5C modification patterns differ in clinicopathological features, genetic heterogeneity, patient prognosis, and treatment responses of ccRCC. An elevated m5C score, characterized by malignant biologic processes of tumor cells and suppression of immunity response, implies an immune-desert TME phenotype and is associated with dismal prognosis of ccRCC. Activation of exhausted T cells and effective immune infiltration were observed in the low m5C score cluster, reflecting a noninflamed and immune-excluded TME phenotype with favorable survival and better responses to immunotherapy. Together, these findings provide insights into the regulation mechanisms of DNA m5C methylation modification patterns on the tumor immune microenvironment. Comprehensive assessment of tumor m5C modification patterns may enhance our understanding of TME cell-infiltrating characterizations and help establish precision immunotherapy strategies for individual ccRCC patients.

Interferon-gamma (IFN-γ) has a complex role in modulating the tumor microenvironment (TME) during renal cell carcinoma (RCC) development. To define the role of IFN-γ response genes in RCC progression, we characterized the differential gene expression, prognostic implications, and DNA variation profiles of selected IFN-γ response signatures, which exhibited a significant hazard ratio for the overall survival (OS) and progression-free survival (PFS) of papillary, chromophobia, and clear cell RCC (ccRCC) patients (n = 944). Prognostic nomograms were constructed to predict the outcomes for ccRCC patients, highlighting the prognostic implications of RANBP2-type and C3HC4-type zinc finger containing 1 (RBCK1). Interestingly, large-scale pan-cancer samples (n = 12,521) and three single-cell RNA datasets revealed that RBCK1 showed markedly differential expression between cancer and normal tissues and significantly correlated with tumor-infiltrating immune cells, tumor purity, and immune checkpoint molecules, such as PD-L1, CTLA-4, LAG-3, and TIGIT in pan-cancer samples. Notably, the TIDE score was significantly higher in the RBCK1high group compared with the RBCK1low group in both ccRCC and RCC cohorts. Besides, immunohistochemistry staining showed significantly elevated RBCK1 expression in tumors (n = 50) compared with kidney samples (n = 40) from a real-world cohort, Fudan University Shanghai Cancer Center (FUSCC, Shanghai). After RBCK1 expression was confirmed in ccRCC, we found a significantly decreased number of infiltrating CD4+ T cells, CD4+ FOXP3+ Treg cells, M1 macrophages, and CD56bight/dim NK cells in the immune-cold RBCK1high group. In addition to the distinct heterogeneous immune microenvironment, the increased expression of RBCK1 predicted a prominently worse prognosis than the RBCK1low group for 232 ccRCC patients in the FUSCC proteomic cohort. Furthermore, after transfected with siRNA in human ccRCC cells, extraordinarily decreased cell proliferation, migration capacities, and prominently elevated apoptosis tumor cell proportion were found in the siRNA groups compared with the negative control group. In conclusion, this study identified IFN-γ response clusters, which might be used to improve the prognostic accuracy of immune contexture in the ccRCC microenvironment. Immune-cold RBCK1high patients have pro-tumorigenic immune infiltration and significantly worse outcomes than RBCK1low patients based on results from multi-omics to real-world data. Our discovery of novel independent prognostic indicators for RCC highlights the association between tumor alterations and immune phenotype.

Myricetin is a type of natural flavonol known for its anticancer activity. However, the molecular mechanism of myricetin in anti-hepatocellular carcinoma (HCC) is not well defined. Previous studies indicated that downregulation of membrane-associated RING-CH finger protein 1 (MARCH1) contributed to the treatment of a variety of cancers. Whether the anticancer property of myricetin is associated with MARCH1 expression remains to be investigated. This research explored the anti-HCC mechanism of myricetin. Our results indicate that myricetin induces autophagy and arrests cell cycle at the G2/M phase to suppress the proliferation of HCC cells by downregulating MARCH1. Myricetin reduces MARCH1 protein in Hep3B and HepG2 cells. Interestingly, myricetin upregulates the MARCH1 mRNA level in Hep3B cells but downregulates it in HepG2 cells. The knockdown of MARCH1 by siRNAs (small interfering RNAs) decreases the phosphorylated p38 MAPK (p-p38 MAPK) and Stat3 (p-Stat3), and inhibits HCC cell viability. Moreover, myricetin inhibits p38 MAPK and Stat3 signaling pathways by downregulating MARCH1 to repress HCC growth both in vitro and in vivo. Bafilomycin A1 (BafA1), an autophagy inhibitor, has synergetic effect with myricetin to inhibit HCC growth. Taken together, our results reveal that myricetin inhibits the proliferation of HCC cells by inhibiting MARCH1-regulated p38 MAPK and Stat3 signaling pathways. This research provides a new molecular mechanism for myricetin in anti-HCC and suggests that targeting MARCH1 could be a novel treatment strategy in developing anticancer therapeutics.

BackgroundThis study aims to establish an N6-methyladenosine (m6A) RNA methylation regulators-mediated methylation model and explore its role in predicting prognostic accuracy of immune contexture and characterizations of clear cell renal cell carcinoma (ccRCC).MethodsThe m6A modification subclasses (m6AMS) were identified by unsupervised cluster analysis and three clusters were determined by consensus clustering algorithm in a discovering cohort. Testing and real-world validation cohorts were used to identify predictive responses for immune checkpoint therapies (ICTs) of m6AMS.ResultsPrognostic implications landscape of m6A regulators in cancers and its differential expression levels in ccRCC patients were identified. Based on discovering cohort, ccRCC were automatically divided into three m6AMS, and cluster 3 showed significant worse survival than cluster 1/2. Importantly, it was found that the immune checkpoint molecules expression was significantly elevated in cluster 3. Besides, m6A scoreLow group (cluster 1&2) have significantly elevated TIDE score compared with m6A scoreHigh group (cluster 3). There was conspicuous tertiary lymphoid tissue, aggressive phenotype, elevated glycolysis, expression of PD-L1, abundance of CD8+ T cells, CD4+ FOXP3+ Treg cells and TCRn immune cells infiltration in the high m6A score group. Interestingly, there are significantly increased patients with clinical benefit in m6A scoreHigh group in 368 patients receiving ICTs from testing IMvigor210 (n = 292) and validation FUSCC (n = 55) cohorts.ConclusionOur discovery highlights the relationship between tumor epigenetic heterogeneity and immune contexture. Immune-rejection cluster 3 has pro-tumorigenic immune infiltration, and shows significant clinical benefits for ccRCC patients receiving ICTs, enabling patient selection for future clinical treatment.

Promotion of apoptosis and suppression of proliferation in tumor cells are popular strategies for developing anticancer drugs. Sinomenine (SIN), a plant-derived alkaloid, displays antitumor activity. However, the mechanism of action of SIN against hepatocellular carcinoma (HCC) is unclear. Herein, several molecular technologies, such as Western Blotting, qRT-PCR, flow cytometry, and gene knockdown were applied to explore the role and mechanism of action of SIN in the treatment of HCC. It was found that SIN arrests HCC cell cycle at G0/G1 phase, induces apoptosis, and suppresses proliferation of HCC cells via down-regulating the expression of membrane-associated RING-CH finger protein 1 (MARCH1). Moreover, SIN induces cell death and growth inhibition through AMPK/STAT3 signaling pathway. MARCH1 expression was silenced by siRNA to explore its involvement in the regulation of AMPK/STAT3 signaling pathway. Silencing MARCH1 caused down-regulation of phosphorylation of AMPK, STAT3 and decreased cell viability and function. Our results suggested that SIN inhibits proliferation and promotes apoptosis of HCC cells by MARCH1-mediated AMPK/STAT3 signaling pathway. This study provides new support for SIN as a clinical anticancer drug and illustrates that targeting MARCH1 could be a novel treatment strategy in developing anticancer therapeutics.

Globally, many terrestrial ecosystems are experiencing a rapid loss of biodiversity. Continued improvements in our understanding of interrelationships between plant diversity and soil microbes are critical to address the concern over the consequences of the decline in biodiversity on ecosystem functioning and services. By removing forbs, or grasses, or, to an extreme scenario, both forbs and grasses in a steppe vegetation in Inner Mongolia, we studied how plant functional group (PFG) loss affects soil microbial community composition using phospholipid fatty acid analysis (PLFA) and litter decomposition using a litter-bag method. PFG loss significantly decreased above- and below-ground plant biomass, soil microbial biomass carbon (SMBC) and nitrogen (SMBN), but had no effect on the ratio of SMBC to SMBN. Although the ratio of fungal to bacterial PLFAs remained unaffected, PFG loss significantly reduced the amount of bacterial, fungal, and total PLFAs. PFG loss decreased litter monthly mass loss and decay constant, and such decrease was significant when both forbs and grasses were removed. Our results provide robust evidence that PFG loss in grassland ecosystem can lead to a rapid response of soil microbial activity which may affect litter decomposition and soil nutrient cycling, suggesting that the assessment of plant–microbe interactions in soils is an integral component of ecosystem response to biodiversity loss.

Hybrid oysters often show heterosis in growth rate, weight, survival and adaptability to extremes of salinity. Oysters have also been used as model organisms to study the evolution of host-defense system. To gain comprehensive knowledge about various physiological processes in hybrid oysters under low salinity stress, we performed transcriptomic analysis of gill tissue of Crassostrea sikamea ♀ × Crassostrea angulata♂ hybrid using the deep-sequencing platform Illumina HiSeq. We exploited the high-throughput technique to delineate differentially expressed genes (DEGs) in oysters maintained in hypotonic conditions. A total of 199,391 high quality unigenes, with average length of 644 bp, were generated. Of these 35 and 31 genes showed up- and down-regulation, respectively. Functional categorization and pathway analysis of these DEGs revealed enrichment for immune mechanism, apoptosis, energy metabolism and osmoregulation under low salinity stress. The expression patterns of 41 DEGs in hybrids and their parental species were further analyzed by quantitative real-time PCR (qRT-PCR). This study will serve as a platform for subsequent gene expression analysis regarding environmental stress. Our findings will also provide valuable information about gene expression to better understand the immune mechanism, apoptosis, energy metabolism and osmoregulation in hybrid oysters under low salinity stress.

Although semi-arid and arid regions account for about 40% of terrestrial surface of the Earth and contain approximately 10% of the global soil organic carbon stock, our understanding of soil organic carbon dynamics in these regions is limited.A field experiment was conducted to compare soil organic carbon dynamics between a perennial grass community dominated by Cleistogenes squarrosa and an adjacent shrub community co-dominated by Reaumuria soongorica and Haloxylon ammodendron, two typical plant life forms in arid ecosystems of saline-alkaline arid regions in northwestern China during the growing season 2010. We found that both fine root biomass and necromass in two life forms varied greatly during the growing season. Annual fine root production in the perennial grasses was 45.6% significantly higher than in the shrubs, and fine root turnover rates were 2.52 and 2.17 yr(-1) for the perennial grasses and the shrubs, respectively. Floor mass was significantly higher in the perennial grasses than in the shrubs due to the decomposition rate of leaf litter in the perennial grasses was 61.8% lower than in the shrubs even though no significance was detected in litterfall production. Soil microbial biomass and activity demonstrated a strong seasonal variation with larger values in May and September and minimum values in the dry month of July. Observed higher soil organic carbon stocks in the perennial grasses (1.32 Kg C m(-2)) than in the shrubs (1.12 Kg C m(-2)) might be attributed to both greater inputs of poor quality litter that is relatively resistant to decay and the lower ability of microorganism to decompose these organic matter.Our results suggest that the perennial grasses might accumulate more soil organic carbon with time than the shrubs because of larger amounts of inputs from litter and slower return of carbon through decomposition.

Abstract Background This study aims to validate a hypothesized model of the relationships between optimism, resilience, coping, parenting stress, perceived social support, and anxiety, as well as their influencing factors and consequences among caregivers of children diagnosed with cleft lip and/or palate (CL/P). Methods A hypothesized model was proposed based on diathesis-stress theoretical model and existing literature. A convenience sample of two CL/P treatment centers in China was collected between April 2019 and July 2020. Various well-validated instruments were used to collect 248 valid data points. Descriptive statistics and correlations were computed to assess model fitness and path analysis was performed to investigate direct and indirect effects. Results The results revealed that the average level of anxiety among caregivers of children suffering from CL/P was 3.00 (0.00,6.00). The fitness of the modified path model was evaluated using various measures, including χ2/df = 1.806, GFI = 0.989, RFI = 0.916, IFI = 0.990, CFI = 0.989, TLI = 0.960, RMSEA = 0.0570. Parenting stress had the greatest direct associated with anxiety, followed by perceived social support. Moreover, indirect relations to anxiety were observed for optimism, resilience, and coping, which were mediated through perceived social support and parenting stress. Conclusion In summary, this work suggests that both parenting stress and perceived social support in caregivers of children suffering from CL/P are associated with anxiety. Therefore, developing effective intervention approaches targeting key factors might potentially instrumental to alleviate anxiety.