摘要：
A novel peptide attenuating skeletal muscle atrophy was prepared, identified, screened from corn and its molecular mechanism was explored using two-step enzymatic hydrolysis, molecular docking, and sarcopenia mice model. The results showed that the DPPH free radical scavenging rate of corn peptides (CPs) was 45.20 % under the optimum preparation conditions. Fifty-one peptide fragments were identified from CPs, among which QQPIVGGA, QYQLPSY, LQQQLL, and LQQQQL presented superior affinity with mTORC1 and FOXO in molecular docking. LQQQLL (0.02 mM) significantly increased the proliferative activity of senescent C2C12 cells by 41.67 % compared with the model group ( P < 0.05), showing the potential to attenuate skeletal muscle atrophy. The sarcopenia mice model results indicated that CPs and LQQQLL significantly improved the content of total superoxide dismutase (T-SOD), skeletal muscle mass index (SMI), and decreased the level of malondialdehyde (MDA), tumor necrosis factor (TNF)-α, muscle atrophy protein Fbox-1 (Atrogin-1), and 8-hydroxydeoxyguanosine (8-OHdG) ( P < 0.05). CPs and LQQQLL also markedly increased the cross-sectional muscle area and the relative content of type II muscle fibers in sarcopenia mice. Additionally, CPs and LQQQLL significantly up-regulated the expression levels of PI3K, AKT and mTOR proteins ( P < 0.05), reduced the proliferation of Proteobacteria , Actinobacteriota , Desulfobacterota, and Staphylococcus and promoted the proliferation of Bacteroidota, and Lactobacillus . In conclusion, CPs and LQQQLL could activate the PI3K/AKT/mTOR signaling pathway, and reduce the proliferation of pathogens and promote the proliferation of intestinal microorganisms, thus improving the attenuation of skeletal muscle.
A novel peptide attenuating skeletal muscle atrophy was prepared, identified, screened from corn and its molecular mechanism was explored using two-step enzymatic hydrolysis, molecular docking, and sarcopenia mice model. The results showed that the DPPH free radical scavenging rate of corn peptides (CPs) was 45.20 % under the optimum preparation conditions. Fifty-one peptide fragments were identified from CPs, among which QQPIVGGA, QYQLPSY, LQQQLL, and LQQQQL presented superior affinity with mTORC1 and FOXO in molecular docking. LQQQLL (0.02 mM) significantly increased the proliferative activity of senescent C2C12 cells by 41.67 % compared with the model group ( P < 0.05), showing the potential to attenuate skeletal muscle atrophy. The sarcopenia mice model results indicated that CPs and LQQQLL significantly improved the content of total superoxide dismutase (T-SOD), skeletal muscle mass index (SMI), and decreased the level of malondialdehyde (MDA), tumor necrosis factor (TNF)-α, muscle atrophy protein Fbox-1 (Atrogin-1), and 8-hydroxydeoxyguanosine (8-OHdG) ( P < 0.05). CPs and LQQQLL also markedly increased the cross-sectional muscle area and the relative content of type II muscle fibers in sarcopenia mice. Additionally, CPs and LQQQLL significantly up-regulated the expression levels of PI3K, AKT and mTOR proteins ( P < 0.05), reduced the proliferation of Proteobacteria , Actinobacteriota , Desulfobacterota, and Staphylococcus and promoted the proliferation of Bacteroidota, and Lactobacillus . In conclusion, CPs and LQQQLL could activate the PI3K/AKT/mTOR signaling pathway, and reduce the proliferation of pathogens and promote the proliferation of intestinal microorganisms, thus improving the attenuation of skeletal muscle.
摘要：
Background: Allergic rhinitis (AR) is a prevalent allergic disorder characterized by a complex pathogenesis. Drawing on traditional Chinese medicine theory and contemporary pharmacological principles, this study developed an inhalation-based herbal formulation, ZHW, to explore a novel non-invasive therapeutic approach. Objective: To investigate the therapeutic effects of ZHW on AR and elucidate its underlying mechanisms and potential targets through an integrated analysis of network pharmacology and proteomics. Materials and Methods: The volatile components of ZHW were analyzed by gas chromatography-mass spectrometry (GC-MS). The mouse model of AR was induced by OVA sensitization. The therapeutic efficacy of ZHW was assessed based on nasal symptom scores, histopathological examination, and inflammatory cytokine levels. Furthermore, the underlying mechanisms and potential targets of ZHW were investigated through integrated network pharmacology and proteomics analyses. Results: GC-MS analysis identified 39 bioactive compounds in ZHW. Inhalation treatment with ZHW demonstrated significant anti-allergic effects in OVA-sensitized mice, as evidenced by (1) reduced sneezing frequency and nasal rubbing behaviors; (2) decreased serum levels of IL-4, histamine, and OVA-specific IgE; (3) attenuated IL-4 concentrations in both nasal lavage fluid and lung tissue; (4) diminished nasal mucosal thickening; and (5) suppression of inflammatory cell infiltration. Integrated network pharmacology and proteomics analyses indicated that ZHW's therapeutic effects were mediated through the modulation of multiple pathways, including the PI3K-Akt signaling pathway, the B cell receptor signaling pathway, oxidative phosphorylation, and the FcεRI signaling pathway. Key molecular targets involved Rac1, MAPK1, and SYK. Molecular docking simulations revealed strong binding affinities between ZHW's primary bioactive constituents (linalool, levomenthol, linoleic acid, Linoelaidic acid, and n-Valeric acid cis-3-hexenyl ester) and these target proteins. Conclusions: The herbal formulation ZHW demonstrates significant efficacy in alleviating allergic rhinitis symptoms through multi-target modulation of key signaling pathways, including PI3K-Akt- and FcεRI-mediated inflammatory responses. These findings substantiate ZHW's therapeutic potential as a novel, non-invasive treatment for AR and provide a strong basis for the development of new AR therapies. Future clinical development will require systematic safety evaluation to ensure optimal therapeutic outcomes.
摘要：
Lotus root is a widely popular aquatic vegetable with edible and medicinal values. Here we report the structure and lipid-lowering activity of two lotus root polysaccharides LRW (lotus root polysaccharide by water extraction) and LRA (lotus root polysaccharide by alkali extraction), that were extracted by aqueous and alkaline solution respectively. The results showed that the yield of polysaccharide from lotus root could be significantly improved by alkali extraction. Basic composition and structural characterization showed that the total sugar contents of LRW and LRA were 96.83 % and 73.66 %, and the molecular weights were 2.464 × 10 5 Da and 1.727 × 10 5 Da, respectively. LRW and LRA had the similar structure that the main backbone consisted of →4)-α-D-Glcp-(1→ with branches at C-6 site. Both LRW and LRA could scavenge DPPH and hydroxyl radicals effectively, and have strong adsorption capacity to cholate salts in a concentration-dependent manner. In HepG2 cells, LRW and LRA inhibited the accumulation of lipid droplets induced by oleic acid, and increased the activity of T-SOD and CAT, meanwhile, reduced the level of MDA, TC and TG, showing good lipid-lowering activity. In comparison, the lipid-lowering effect of LRA was better than that of LRW. In addition, gene sequencing and RT-PCR showed that AMPK, ACC, PPARα and CPT-1 were essential for LRA to exert a lipid-lowering effect. This study provides a theoretical basis for the extraction and lipid-lowering application of lotus root polysaccharides.
Lotus root is a widely popular aquatic vegetable with edible and medicinal values. Here we report the structure and lipid-lowering activity of two lotus root polysaccharides LRW (lotus root polysaccharide by water extraction) and LRA (lotus root polysaccharide by alkali extraction), that were extracted by aqueous and alkaline solution respectively. The results showed that the yield of polysaccharide from lotus root could be significantly improved by alkali extraction. Basic composition and structural characterization showed that the total sugar contents of LRW and LRA were 96.83 % and 73.66 %, and the molecular weights were 2.464 × 10 5 Da and 1.727 × 10 5 Da, respectively. LRW and LRA had the similar structure that the main backbone consisted of →4)-α-D-Glcp-(1→ with branches at C-6 site. Both LRW and LRA could scavenge DPPH and hydroxyl radicals effectively, and have strong adsorption capacity to cholate salts in a concentration-dependent manner. In HepG2 cells, LRW and LRA inhibited the accumulation of lipid droplets induced by oleic acid, and increased the activity of T-SOD and CAT, meanwhile, reduced the level of MDA, TC and TG, showing good lipid-lowering activity. In comparison, the lipid-lowering effect of LRA was better than that of LRW. In addition, gene sequencing and RT-PCR showed that AMPK, ACC, PPARα and CPT-1 were essential for LRA to exert a lipid-lowering effect. This study provides a theoretical basis for the extraction and lipid-lowering application of lotus root polysaccharides.
作者机构：
[Deng, Ying; Wang, Haoqian; Xia, Shutao; Zhang, Tian] Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, People's Republic of China;[Wu, Yiling; Xia, Shutao; Zhang, Tian] Peng Cheng Laboratory, Shenzhen, China;[Wang, Wentao; Zhao, Zhe] Zhengzhou KingMed Center for Clinical Laboratory, Zhengzhou, Henan, People's Republic of China;[Liao, Weifang] College of life science and technology, Wuhan Polytechnic University, Wuhan, People's Republic of China. leesalwf89@126.com;[Liao, Weijie] Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, People's Republic of China. liaoweijie@szu.edu.cn
关键词：
Machine learning models;Metatranscriptomic sequencing;Patients in severe condition;Prompt diagnosis;Pulmonary infections
摘要：
The prompt diagnosis of pulmonary infections with unknown etiology in patients in severe condition remains a challenge due to the lack of rapid and effective diagnostic methods. While metatranscriptomic sequencing offers a powerful approach, its clinical utility is often limited by issues of timeliness. In this study, we conducted metatranscriptomic sequencing on bronchoalveolar lavage fluid (BALF) collected from critically ill, severely ill, and ICU patients. Based on microbial detection results, patients were classified into four types: negative, bacterial infection, viral infection, and fungal infection. To identify host gene expression signatures associated with infection, we screened characteristic genes from human metatranscriptomic data by comparing 70% of patients with confirmed infections vs. non-infections. Leveraging these characteristic genes, we constructed classification sub-models employing 13 types of machine learning algorithms, and we further integrated these sub-models into stacking-based ensemble models with Lasso regression, resulting in diagnostic models that required only a small set of gene expression inputs. The average performance of five-fold cross-validation demonstrated high diagnostic accuracy: distinguishing infection from non-infection (AUC = 0.984), bacterial infection from non-bacterial infection (AUC = 0.98), and viral infection from non- viral infection (AUC = 0.98). Test cohorts' results demonstrated the method's high diagnostic accuracy consistency with metatranscriptomic sequencing in discerning patient infection status (AUC = 0.865) and the type of infection (viral: AUC = 0.934, bacterial: AUC = 0.871). Our study presented a rapid and inexpensive adjunctive diagnostic strategy that achieves diagnostic accuracy comparable to metatranscriptomic sequencing, enabling timely identification of both infection status and type in pulmonary infections.
作者：
Jie Wu;Ying Sun*;Xueyu Jiang;Kaidi Peng;Hongxun Wang;...
期刊：
LWT,2025年:118391 ISSN：0023-6438
通讯作者：
Ying Sun
作者机构：
[Jie Wu; Ying Sun; Xueyu Jiang; Kaidi Peng; Yang Yi] College of Food Science and Engineering/Hubei Key Laboratory for Processing and Transformation of Agricultural Products, Wuhan Polytechnic University, Wuhan 430023, China;[Hongxun Wang] College of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, China
通讯机构：
[Ying Sun] C;College of Food Science and Engineering/Hubei Key Laboratory for Processing and Transformation of Agricultural Products, Wuhan Polytechnic University, Wuhan 430023, China
摘要：
In this study, ultrafine grinding whole lotus root powder (LR) was combined with whey protein (WP) to prepare Pickering emulsions. The effects of LR/WP ratio, solid content and oil-water ratio on the stability of the emulsion were investigated, and the underlying mechanisms were explored. Results indicated that ultrafine grinding modification and WP combination significantly enhanced the surface hydrophobicity of LR. When the ratio of LR/WP ranged from 5:1 to 15:1, the solid content was ≥ 6 wt%, and the oil-water ratio spanned from 2:1 to 4:1, long-term stable emulsions could be formed with small particle sizes and high absolute values of zeta potential. Meanwhile, the combination of LR and WP enabled the preparation of various stable emulsions with different oil contents (conventional emulsions and high internal-phase emulsions), showing high adaptability. The exploration of the stability mechanism demonstrated that LR/WP complex uniformly covered the surface of the droplets, forming a dense interface layer, and the emulsion system presented an elastic-dominant gel-like network structure, thereby ensuring the stability of the emulsion system. This study elucidated the influence of the interfacial properties and stability of LR/WP Pickering emulsions, providing theoretical basis and technical support for the development of plant-based whole component emulsions.
In this study, ultrafine grinding whole lotus root powder (LR) was combined with whey protein (WP) to prepare Pickering emulsions. The effects of LR/WP ratio, solid content and oil-water ratio on the stability of the emulsion were investigated, and the underlying mechanisms were explored. Results indicated that ultrafine grinding modification and WP combination significantly enhanced the surface hydrophobicity of LR. When the ratio of LR/WP ranged from 5:1 to 15:1, the solid content was ≥ 6 wt%, and the oil-water ratio spanned from 2:1 to 4:1, long-term stable emulsions could be formed with small particle sizes and high absolute values of zeta potential. Meanwhile, the combination of LR and WP enabled the preparation of various stable emulsions with different oil contents (conventional emulsions and high internal-phase emulsions), showing high adaptability. The exploration of the stability mechanism demonstrated that LR/WP complex uniformly covered the surface of the droplets, forming a dense interface layer, and the emulsion system presented an elastic-dominant gel-like network structure, thereby ensuring the stability of the emulsion system. This study elucidated the influence of the interfacial properties and stability of LR/WP Pickering emulsions, providing theoretical basis and technical support for the development of plant-based whole component emulsions.
摘要：
Ferritin heavy chain 1 (FTH1) is a key iron-storage protein that regulates iron availability, supports immune defense, and prevents iron-induced toxicity. During Mycobacterium tuberculosis (Mtb) infection, macrophages enhance FTH1 expression to sequestrate iron and limit Mtb growth. However, Mtb can exploit the host ferritinophagy pathway to degrade FTH1 and release iron, thereby promoting its survival. Although FTH1 plays an essential role in host-pathogen interaction during Mtb infection, its regulation remains unclear. Previous studies suggest that post-transcriptional mechanism, particularly alternative polyadenylation (APA), are critical in immune responses. We propose that APA, which determines the length of a transcript's 3'UTR, may regulate FTH1 expression during Mtb infection. Our study demonstrates that Mtb induces APA of FTH1 in macrophages, favoring the production of longer isoforms that enhance protein synthesis. Mechanistically, Mtb disrupts the interaction between NUDT21 and CPSF6, impairing NUDT21's ability to bind UGUA motifs in the FTH1 3'UTR, a key step in polyadenylation site selection. Silencing NUDT21 reduces macrophage bactericidal activity against Mtb, highlighting its role in immune defense. These findings reveal a novel Mtb-driven mechanism that enhances FTH1 expression via the NUDT21-mediated APA pathway in macrophages, suggesting that Mtb manipulates this process to promote its survival. This study provides new insights into tuberculosis pathogenesis and points to potential avenues for therapeutic exploration.
通讯机构：
[Yang, JK ; Han, ZG] W;Wuhan Polytech Univ, Coll Life Sci & Technol, Wuhan 430023, Peoples R China.
关键词：
glycoside hydrolase family 10;low-temperature enzymes;molecular dynamics simulation;xylanases
摘要：
This study investigates the biochemical properties of two xylanases, ZgXyn10A and CaXyn10B, which are members of the glycoside hydrolase family 10 (GH10) and originate from the marine Bacteroidetes species Zobellia galactanivorans and Cellulophaga algicola, respectively. Utilizing an auto-induction expression system in Escherichia coli, high-purity recombinant forms of these enzymes were successfully produced. Biochemical assays revealed that ZgXyn10A and CaXyn10B exhibit optimal activities at 40 degrees C and 30 degrees C, respectively, and demonstrate a high sensitivity to temperature fluctuations. Unlike conventional low-temperature enzymes, these xylanases retain only a fraction of their maximal activity at lower temperatures. To gain deeper insights into the structural and functional properties of these marine xylanases, two thermostable GH10 xylanases, TmxB and CoXyn10A, which share comparable amino acid sequence identity with ZgXyn10A and CaXyn10B, were selected for structural comparison. All four marine xylanases share a nearly similar three-dimensional structural topology. Molecular dynamics simulation indicated a striking difference in structural fluctuations between the low-temperature and thermostable xylanases, as evidenced by the distinct root mean square deviation values. Moreover, root mean square fluctuation analysis specifically identified the beta 3-alpha 3 and beta 7-alpha 7 loop regions within the substrate-binding cleft as crucial determinants of the temperature characteristics of these GH10 xylanases. Our findings establish loop dynamics as a key evolutionary driver in the thermal adaptation of GH10 xylanases and propose a loop engineering strategy for the development of industrial biocatalysts with tailored temperature responses, particularly for lignocellulosic biomass processing under moderate thermal conditions.
通讯机构：
[Guo, DJ ] W;Wuhan Polytech Univ, Coll Food Sci & Engn, Wuhan 430040, Peoples R China.
关键词：
carbonyls;fatty acids;sous vide;TBARS
摘要：
The laws and interrelationships of lipid and protein oxidation in duck meat during sous vide cooking (SVC) are still unclear. In this study, the oxidation mechanisms of the muscle of duck legs during SVC were investigated from the aspects of fatty acid (FA) profile, antioxidant system activity, enzyme activity, and iron element. Lipid oxidation predominantly occurred within the initial 0-2 h of the SVC process, during which the Thiobarbituric acid reactive substances (TBARS) value escalated to 10-fold of its initial level. Protein oxidation was mainly observed between 0 and 4 h, with the carbonyl content increasing by 23.3%. The FAs profile underwent changes primarily within the initial 0-2 h, and then their proportions continued to evolve in a manner that was favorable for oxidative stability. Following 4 h of SVC, the scavenging rates of <middle dot>OH and DPPH<middle dot> free radicals decreased to the lowest, decreasing by 92.9% and 45.6%. As cooking time increased, lipoxygenase activity increased by more than six times, while Total superoxide dismutase (T-SOD) and GSH-Px activity decreased by 31.7% and 75.2%, respectively. The heme iron content was decreased by 26.1% after 8 h of SV treatment. These data can provide a theoretical foundation for optimizing parameters of the SVC process and improving the oxidative stability of SVC meat products.
摘要：
Background: Programmed cell death-related genes (PCDRGs) have been reported to play an important role in diagnosis, treatment and immunity regarding cancer, but their prognostic value and therapeutic potential in acute myeloid leukemia (AML) patients still need to be fully explored. Methods: Cox regression analysis and Least Absolute Shrinkage and Selection Operator (LASSO) analysis were used to identify PCDRGs significantly associated with the prognosis of AML patients. Furthermore, a prognostic risk model for AML patients was constructed based on the selected PCDRGs, and their immune microenvironment and biological pathways were analyzed. Cell experiments ultimately confirmed the potential role of PCDRGs in AML. Results: The results yielded four PCDRGs that were used to develop a prognostic risk model, and the prognostic significance of this model was confirmed using an independent external AML patient cohort. This prognostic risk model provides an independent prognostic risk factor for AML patients. This prognostic feature is related to immune cell infiltration in AML patients. The inhibition of solute carrier family 39 member 14 (SLC39A14) expression enhanced apoptosis and inhibited cell cycle progression in AML cells. Conclusions: This study integrates bioinformatics analysis and cellular experiments to reveal potential gene therapy targets and prognostic gene markers in AML.
摘要：
Previous studies have demonstrated corilagin’s inhibitory effects on the growth of various cancer cells. Given the limited research on corilagin’s impact on ovarian cancer, a particularly deadly gynecological malignancy, this study aimed to investigate corilagin’s influence on A2780 ovarian cancer cell apoptosis and its underlying mechanisms. The goal was to evaluate corilagin’s potential as a therapeutic agent for ovarian cancer. The results of the CCK-8 assay showed that corilagin inhibited the proliferation of A2780 ovarian cancer cells while exhibiting lower toxicity to normal ovarian surface epithelial cells (IOSE-80). We found that corilagin significantly altered the A2780 cell cycle, decreasing the proportion of cells in the G0/G1 and G2/M phases and inducing cell cycle arrest in the S phase. At low concentrations, corilagin induced apoptosis in A2780 cells, accompanied by a decline in mitochondrial membrane potential and calcium influx. Transcriptome sequencing analysis identified differentially expressed apoptosis-related genes in corilagin-treated A2780 cells, primarily within the PI3K-AKT pathway. Furthermore, qPCR and Western blot results confirmed the upregulation of p53 and Bax genes and the downregulation of BCL-2. Corilagin also increased the expression of apoptotic factors caspase-9, caspase-3, PUMA, and cytochrome C, indicating its ability to induce apoptosis. Overall, corilagin effectively inhibited A2780 cell proliferation, induced cell cycle arrest, and triggered apoptosis. Its anti-tumor effect in vitro suggests its potential as a therapeutic agent for ovarian cancer A2780, especially through the PI3K/p53 pathway.
摘要：
The aprBP gene from Bacillus patagoniensis DB-5, encoding a 378-amino-acid alkaline protease, was cloned and expressed in Escherichia coli. The amino acid sequence of APrBP showed 62.8-84.4% identity with the S8 peptidase subtilisin family alkaline proteases reported in the literature. Recombinant APrBP was purified using Ni-NTA affinity chromatography with 45.61% recovery and a homogeneous band was detected at approximately 38 kDa on the SDS-PAGE gel. The optimum temperature of APrBP was 60°C. The presence of 2 mM Ca(2+) significantly enhanced the optimal temperature and thermostability. The enzyme demonstrated optimum activity at pH 12 and maintained high stability at pH 8.0-11.0. Protease activity was stimulated by Mn(2+), Ca(2+), Mg(2+), Ni(2+), TritonX-100, Tween-20 and Tween-80, while completely inactivated by PMSF, EDTA and Cu(2+). The APrBP exhibited good tolerance to oxidizing and reducing agents. Notably, the protease exhibited remarkable stability in 50% (v/v) concentrations of several organic solvents, such as methanol, acetone, glycerol, dimethyl sulfoxide, n-hexane, and ethyl acetate. The APrBP efficiently hydrolyzed natural proteins, demonstrating the highest catalytic efficiency for casein, excellent hydrolysis activity for bovine serum albumin, hemoglobin, and keratin, and favorable hydrolysis ability for whey proteins. Moreover, molecular docking results revealed stable interactions between APrBP and casein, hemoglobin, whey proteins and keratin. This study indicated that APrBP has some useful properties and explored its potential as a bio-additive detergent as well as in utilizing feather waste and whey protein.
摘要：
Objectives The persistence and infectivity of respiratory viruses in cadavers remain poorly characterized, posing significant biosafety risks for forensic and healthcare professionals. This study systematically evaluates the post-mortem stability and transmission potential of SARS-CoV-2, influenza A virus (IAV), and respiratory syncytial virus (RSV) under varying environmental conditions, providing critical insights into viral kinetics.
The persistence and infectivity of respiratory viruses in cadavers remain poorly characterized, posing significant biosafety risks for forensic and healthcare professionals. This study systematically evaluates the post-mortem stability and transmission potential of SARS-CoV-2, influenza A virus (IAV), and respiratory syncytial virus (RSV) under varying environmental conditions, providing critical insights into viral kinetics.
Methods To assess the post-mortem stability of SARS-CoV-2, tissue samples were collected from infected cadavers at 4℃, room temperature (RT, 20-22℃), and 37℃ over a predetermined timeframe. Viral kinetics were analyzed using quantitative assays, while histopathology and immunohistochemistry characterized tissue-specific distribution. Additionally, comparative analyses were conducted both in vitro and in cadaveric tissues to characterize the survival dynamics of IAV and RSV under identical conditions.
To assess the post-mortem stability of SARS-CoV-2, tissue samples were collected from infected cadavers at 4℃, room temperature (RT, 20-22℃), and 37℃ over a predetermined timeframe. Viral kinetics were analyzed using quantitative assays, while histopathology and immunohistochemistry characterized tissue-specific distribution. Additionally, comparative analyses were conducted both in vitro and in cadaveric tissues to characterize the survival dynamics of IAV and RSV under identical conditions.
Results SARS-CoV-2 exhibited prolonged post-mortem infectivity, persisting for up to 5 days at RT and 37℃ and over 7 days at 4℃, with the highest risk of transmission occurring within the first 72 hours at RT and 24 hours at 37℃. In contrast, RSV remained viable for 1-2 days, while IAV persisted for only a few hours post-mortem. Viral decay rates were temperature-dependent and varied across tissues, demonstrating distinct post-mortem survival kinetics.
SARS-CoV-2 exhibited prolonged post-mortem infectivity, persisting for up to 5 days at RT and 37℃ and over 7 days at 4℃, with the highest risk of transmission occurring within the first 72 hours at RT and 24 hours at 37℃. In contrast, RSV remained viable for 1-2 days, while IAV persisted for only a few hours post-mortem. Viral decay rates were temperature-dependent and varied across tissues, demonstrating distinct post-mortem survival kinetics.
Conclusions This study presents the first comprehensive analysis of viral persistence in cadavers, revealing prolonged SARS-CoV-2 stability compared to IAV and RSV. These findings underscore the need for enhanced post-mortem biosafety protocols to mitigate occupational exposure risks in forensic and clinical settings. By elucidating viral decay dynamics across environmental conditions, this research establishes a critical foundation for infection control strategies, informing biosafety policies for emerging respiratory pathogens.
This study presents the first comprehensive analysis of viral persistence in cadavers, revealing prolonged SARS-CoV-2 stability compared to IAV and RSV. These findings underscore the need for enhanced post-mortem biosafety protocols to mitigate occupational exposure risks in forensic and clinical settings. By elucidating viral decay dynamics across environmental conditions, this research establishes a critical foundation for infection control strategies, informing biosafety policies for emerging respiratory pathogens.
Data availability All data are available in the main text or the supplementary materials .
All data are available in the main text or the supplementary materials .
摘要：
BACKGROUND: Liquid-liquid phase separation has been reported to be closely related to the development and occurrence of tumors, but its role in osteosarcoma remains unexplored. METHODS: For the first time, based on liquid-liquid phase separation related genes (LLPSRGs), a prognostic model was constructed through multivariate Cox regression. The model was validated on an independent dataset. The patients were grouped based on the median risk score. Simultaneously conduct immune infiltration analysis, enrichment analysis and drug sensitivity evaluation. RESULTS: We have identified for the first time 3 LLPSRGs associated with osteosarcoma prognosis. A prognostic model was constructed based on LLPSRGs and validated in an independent dataset. The signature is an independent risk factor for osteosarcoma patients, the signature was associated with the tumor microenvironment. CONCLUSIONS: we conducted a comprehensive analysis of LLPSRGs in osteosarcoma patients for the first time and identified 3 LLPSRGs that may be associated with the prognosis of osteosarcoma patients.
摘要：
Background Proteins which are important biomarkers for disease diagnosis, prognosis and treatment are often present at very low concentrations in biological samples. Measuring these low-level proteins is important yet challenging. Conventional enzyme-linked immunoassays (ELISAs) usually lack the sensitive required. While ultrasensitive detection methods, such as digital ELISA have recently significantly advanced in sensitivity, they usually face high-cost of using paramagnetic beads or require complicated substrate fabrication, limiting their accessibility for large-scale screening or application in resource-limited settings. Therefore, there is a pressing need to develop cost-effective detection methods with ultrahigh sensitivity to accurately detecting low-level protein biomarkers.
Proteins which are important biomarkers for disease diagnosis, prognosis and treatment are often present at very low concentrations in biological samples. Measuring these low-level proteins is important yet challenging. Conventional enzyme-linked immunoassays (ELISAs) usually lack the sensitive required. While ultrasensitive detection methods, such as digital ELISA have recently significantly advanced in sensitivity, they usually face high-cost of using paramagnetic beads or require complicated substrate fabrication, limiting their accessibility for large-scale screening or application in resource-limited settings. Therefore, there is a pressing need to develop cost-effective detection methods with ultrahigh sensitivity to accurately detecting low-level protein biomarkers.
Results In the study, we developed a cost-effective and easily prepared PDMS microwell array platform for bead-free digital enzyme-linked immunoassay (ELISA) to achieve ultrasensitive protein detection. Both the immunocomplexes formation and separation process, which are two key steps in digital ELISA occur within the PDMS microwells, eliminating the need for paramagnetic beads and complex bead-separation systems. This approach significantly reduces detection costs compared to the widely used single-molecule arrays (SiMoAs)-based digital ELISA, which relies on large quantities of paramagnetic beads for immunocomplexes immobilization and intricate microwell array chips for immunocomplexes separation. Here, the PDMS microwell array chip was modified to generate large-scale femtoliter droplet arrays for single-molecule reactions, with minimized droplet evaporation and no cross-contamination. The PDMS microwell array platform was further successfully applied to single-molecule enzyme detection and cytokine analysis, achieving detection limits of 1.57 fM for streptavidin-β-galactosidase (SβG) and 12.3 fg/ml for human interleukin-6.
In the study, we developed a cost-effective and easily prepared PDMS microwell array platform for bead-free digital enzyme-linked immunoassay (ELISA) to achieve ultrasensitive protein detection. Both the immunocomplexes formation and separation process, which are two key steps in digital ELISA occur within the PDMS microwells, eliminating the need for paramagnetic beads and complex bead-separation systems. This approach significantly reduces detection costs compared to the widely used single-molecule arrays (SiMoAs)-based digital ELISA, which relies on large quantities of paramagnetic beads for immunocomplexes immobilization and intricate microwell array chips for immunocomplexes separation. Here, the PDMS microwell array chip was modified to generate large-scale femtoliter droplet arrays for single-molecule reactions, with minimized droplet evaporation and no cross-contamination. The PDMS microwell array platform was further successfully applied to single-molecule enzyme detection and cytokine analysis, achieving detection limits of 1.57 fM for streptavidin-β-galactosidase (SβG) and 12.3 fg/ml for human interleukin-6.
Significance A low-cost PDMS microwell array platform for digital ELISA was developed. The fabrication of PDMS microwell array chip was straightforward by soft lithography, enabling mass production with small batch-to-batch variation. With its cost-effective fabrication process and high detection sensitivity, the PDMS microwell array would be a promising platform for ultrasensitive protein detection, especially in the area of early disease screening.
A low-cost PDMS microwell array platform for digital ELISA was developed. The fabrication of PDMS microwell array chip was straightforward by soft lithography, enabling mass production with small batch-to-batch variation. With its cost-effective fabrication process and high detection sensitivity, the PDMS microwell array would be a promising platform for ultrasensitive protein detection, especially in the area of early disease screening.
通讯机构：
[Min, T ] W;Wuhan Polytech Univ, Coll Food Sci & Engn, Wuhan 430023, Peoples R China.
关键词：
Salicylic Acid;Fresh-cut Chinese water chestnuts;Yellowing;Browning;Phenylpropane metabolic;Transcriptome analysis
摘要：
Fresh-cut Chinese water chestnuts (CWCs) are susceptible to yellowing and browning, which degrades their overall quality and palatability. Therefore, there is an urgent need to develop effective methods and techniques to maintain their color and freshness. This study aimed to analyze the mechanisms underlying the effects of salicylic acid (SA) treatment on the quality deterioration of fresh-cut CWCs during storage. Soaking fresh-cut CWCs in 3.5 mmol of SA for 40 min and subsequently storing them delayed the yellowing of fresh-cut CWCs. This treatment also mitigated weight loss and loss of soluble solids. In addition, SA treatment enhanced the accumulation of total phenolics by promoting the activity of phenylalanine ammonia-lyase (PAL) and concomitantly suppressing the activities of browning-associated enzymes (PPO and POD). Meanwhile, the application of SA increased the activity of antioxidant enzymes such as SOD and CAT while limiting the accumulation of reactive oxygen species (ROS). Moreover, SA treatment delayed the yellowing and browning of fresh-cut CWCs by down-regulating the expression of CwCHS1 , CwCHS2 , CwCHS3 , CwCHI1 , CwCHI2 , CwMYC2 , CwERF5-like , and CwERF069-like and by limiting the synthesis of flavonoids. Finally, SA treatment upregulated the expression of the ERF transcription factors CwAP2/ERF , CwERF1 , CwERF4A , and CwERF4B and concurrently promoted ethylene biosynthesis, which, in turn, accelerated the senescence and microbial growth of fresh-cut CWCs.
Fresh-cut Chinese water chestnuts (CWCs) are susceptible to yellowing and browning, which degrades their overall quality and palatability. Therefore, there is an urgent need to develop effective methods and techniques to maintain their color and freshness. This study aimed to analyze the mechanisms underlying the effects of salicylic acid (SA) treatment on the quality deterioration of fresh-cut CWCs during storage. Soaking fresh-cut CWCs in 3.5 mmol of SA for 40 min and subsequently storing them delayed the yellowing of fresh-cut CWCs. This treatment also mitigated weight loss and loss of soluble solids. In addition, SA treatment enhanced the accumulation of total phenolics by promoting the activity of phenylalanine ammonia-lyase (PAL) and concomitantly suppressing the activities of browning-associated enzymes (PPO and POD). Meanwhile, the application of SA increased the activity of antioxidant enzymes such as SOD and CAT while limiting the accumulation of reactive oxygen species (ROS). Moreover, SA treatment delayed the yellowing and browning of fresh-cut CWCs by down-regulating the expression of CwCHS1 , CwCHS2 , CwCHS3 , CwCHI1 , CwCHI2 , CwMYC2 , CwERF5-like , and CwERF069-like and by limiting the synthesis of flavonoids. Finally, SA treatment upregulated the expression of the ERF transcription factors CwAP2/ERF , CwERF1 , CwERF4A , and CwERF4B and concurrently promoted ethylene biosynthesis, which, in turn, accelerated the senescence and microbial growth of fresh-cut CWCs.
摘要：
BACKGROUND: Diseases are often caused by multiple factors, angiogenesis-related genes (ARGs) have been shown to be associated with cancer, however, their role in colon cancer had not been fully explored. This study investigated potential biomarkers based on ARGs to improve prognosis and treatment effect in colon cancer. METHODS: ARGs associated with colon cancer prognosis were identified using Cox regression analysis and LASSO analysis. Furthermore, a prognostic model was constructed in colon cancer based on the 3 ARGs, and its biological function were analyzed. We evaluated the differences in tumor immune microenvironment based on prognostic signature. Finally, cell experiments confirmed the function of genes in colon cancer. RESULTS: The prognostic value of ARGs in colon cancer patients has been comprehensively analyzed for the first time and identified 3 ARGs with prognostic values. A prognosis risk model was constructed based on 3 ARGs and its prognostic value was validated on an independent external colon cancer dataset. In colon cancer patients, this prognostic feature was an independent risk factor and was significantly correlated with clinical feature information of colon cancer patients. This feature was also related to the immune microenvironment of colon cancer. Cell experiments showed that high expression of TNF Receptor Superfamily Member 1B (TNFRSF1B) significantly promoted apoptosis and inhibited proliferation of colon cancer cells. Therefore, TNFRSF1B may become an important regulatory factor in the progression of colon cancer by participating in intracellular functional regulation. CONCLUSIONS: This study constructed a prognostic risk model based on three ARGs and for the first time discovered that TNFRSF1B may become an important regulatory factor in cancer progression by participating in intracellular functional regulation.
关键词：
water caltrop;origins;nutritional quality;biological activity;antioxidant capacity
摘要：
This study analyzed eight origins of water caltrop: HH; XG; XT; ST; JY; CZ; YN; JX. The evaluations focused on visual appearance, nutritional quality, and bioactive substances. Additionally, the shells and pulp of the JY were examined for the same parameters. The results demonstrated that the JY exhibited the highest total phenolic content (121.74 mg GAE/100 g) and total flavonoid content (196.67 mg GAE/100 g). The XG demonstrated the highest water content (87.35%) and soluble protein content (15.36 mg/g). JX exhibits the highest total phenolic and flavonoid content, as well as the strongest DPPH radical scavenging rate in the fruit pulp, indicating its superior biological activity and antioxidant capacity compared to water caltrop from other regions. In addition, JX has the highest soluble solids and sugar content in fruit pulp, indicating a sweeter taste. The YN exhibited the highest pulp starch and lowest water content. Principal component analysis revealed that the pulp of the ST and the shell of the JY scored the highest. These findings provide valuable insights for evaluating and processing the nutritional quality of water caltrop from different sources and provide a theoretical basis for consumers to choose water caltrop according to their needs.
作者机构：
[Yang, Jiangke; Wang, Shiqing; Zheng, Yanli; Deng, Yuhui; Hu, Ping; Lei, Lei] Wuhan Polytech Univ, Coll Life Sci & Technol, Wuhan 430023, Peoples R China.;[Chan, Zhuhua; Hu, Ping] Marine Biol Resources Dev & Utilizat Engn Technol, TIO, MNR, Xiamen 361005, Fujian, Peoples R China.;[Peng, Wenfang; Peng, WF; Xue, Qingxin; Li, Jiaxin] Hubei Univ, Hubei Engn Res Ctr Bioenzyme Catalysis, Environm Microbial Technol Ctr Hubei Prov, Sch Life Sci,State Key Lab Biocatalysis & Enzyme E, Wuhan 430062, Peoples R China.
通讯机构：
[Yang, JK ] W;[Peng, WF ] H;[Chan, ZH ] M;Wuhan Polytech Univ, Coll Life Sci & Technol, Wuhan 430023, Peoples R China.;Marine Biol Resources Dev & Utilizat Engn Technol, TIO, MNR, Xiamen 361005, Fujian, Peoples R China.
关键词：
CRISPR-Cas9;High-yield and industrial-scale;Kluyveromyces lactis;Precision strain engineering;Recombinant calf chymosin;Secretory production
摘要：
As an important industrial enzyme, chymosin has been widely used in cheese manufacturing. Fermentation with Kluyveromyces lactis has allowed recombinant chymosin production to fit the growing global demand for cheese consumption; yet improvements can be made to allow for stable and larger-scale production. In this work, various c hymosin p roducing (CP) strains were constructed via targeted chromosomal integration of various copies of a p rochymosin e xpression c assette (PEC) using a CRISPR-Cas9 platform optimized for K. lactis. It enabled the demonstration that chymosin yields could be increased along with gradual chromosomal accumulation of PEC inserts within up to 3 copies. Finally, an optimal CP3i strain was constructed, and with which high yields of recombinant chymosin were attained, reaching ca. 1,200 SU/mL in shake-flask fermentation and ca. 28,000 SU/mL in batch-mode bioreaction, respectively. The activity of the product in milk-curding was observed. These findings provide direction to apply K. lactis-based platforms in the subsequent industrial-scale production of recombinant chymosin.
As an important industrial enzyme, chymosin has been widely used in cheese manufacturing. Fermentation with Kluyveromyces lactis has allowed recombinant chymosin production to fit the growing global demand for cheese consumption; yet improvements can be made to allow for stable and larger-scale production. In this work, various c hymosin p roducing (CP) strains were constructed via targeted chromosomal integration of various copies of a p rochymosin e xpression c assette (PEC) using a CRISPR-Cas9 platform optimized for K. lactis. It enabled the demonstration that chymosin yields could be increased along with gradual chromosomal accumulation of PEC inserts within up to 3 copies. Finally, an optimal CP3i strain was constructed, and with which high yields of recombinant chymosin were attained, reaching ca. 1,200 SU/mL in shake-flask fermentation and ca. 28,000 SU/mL in batch-mode bioreaction, respectively. The activity of the product in milk-curding was observed. These findings provide direction to apply K. lactis-based platforms in the subsequent industrial-scale production of recombinant chymosin.
摘要：
A critical challenge in advancing gene therapy lies in developing safe and efficient gene delivery vectors. While polyethyleneimine (PEI) demonstrates superior DNA condensation capacity as a non-viral vector, its clinical application is hindered by inherent cytotoxicity. To address these challenges, this study presents a novel gene delivery system utilizing natural polyphenol resveratrol (Res) as a cross-linker through its hydroxyl groups capable of interacting with the amino groups in PEI by chemical modification. Three resveratrol-modified PEI (Res-P600, Res-P1800 and Res-P10k) polymers were synthesized and structurally validated by FTIR and 1 H NMR. The gene delivery efficacy of Res-PEI was rigorously examined through in vitro assessments, focusing on DNA condensation capacity, DNA protection ability and cytotoxicity. Concurrently, the particle size, zeta potential, cell uptake efficiency and transfection efficiency of the Res-PEI/DNA polyplexes were quantitatively characterized. Experimental data revealed that Res-PEI/DNA polyplexes effectively shielded genetic material against enzymatic degradation while demonstrating rapid cellular uptake in 293T cells. Cytotoxicity evaluations confirmed Res-PEI's improved biosafety profile relative to conventional PEI polymers. Notably, Res-P10k demonstrated superior transfection efficiency (78.96% at w/w=1) in 293T cells which was closed to lipo6000. Data from HeLa cells align with our original findings in 293T cells, demonstrating broader applicability. Collectively, these findings position Res-PEI polyplexes as a biocompatible and effective alternative for next-generation gene delivery systems.
A critical challenge in advancing gene therapy lies in developing safe and efficient gene delivery vectors. While polyethyleneimine (PEI) demonstrates superior DNA condensation capacity as a non-viral vector, its clinical application is hindered by inherent cytotoxicity. To address these challenges, this study presents a novel gene delivery system utilizing natural polyphenol resveratrol (Res) as a cross-linker through its hydroxyl groups capable of interacting with the amino groups in PEI by chemical modification. Three resveratrol-modified PEI (Res-P600, Res-P1800 and Res-P10k) polymers were synthesized and structurally validated by FTIR and 1 H NMR. The gene delivery efficacy of Res-PEI was rigorously examined through in vitro assessments, focusing on DNA condensation capacity, DNA protection ability and cytotoxicity. Concurrently, the particle size, zeta potential, cell uptake efficiency and transfection efficiency of the Res-PEI/DNA polyplexes were quantitatively characterized. Experimental data revealed that Res-PEI/DNA polyplexes effectively shielded genetic material against enzymatic degradation while demonstrating rapid cellular uptake in 293T cells. Cytotoxicity evaluations confirmed Res-PEI's improved biosafety profile relative to conventional PEI polymers. Notably, Res-P10k demonstrated superior transfection efficiency (78.96% at w/w=1) in 293T cells which was closed to lipo6000. Data from HeLa cells align with our original findings in 293T cells, demonstrating broader applicability. Collectively, these findings position Res-PEI polyplexes as a biocompatible and effective alternative for next-generation gene delivery systems.
