Search Results (170)

Sheng Mai San (SMS), a traditional Chinese medicine formula for treating qi and yin deficiency, is widely used in the management of conditions such as cardiovascular diseases and heatstroke. However, its role in mitigating heat stress (HS)-induced liver injury remains underexplored. In this study, a rat model of HS was established under high-temperature and high-humidity conditions, and SMS was administered as an intervention. The pharmacodynamic effects of SMS were comprehensively evaluated through histopathological examination, detection of heat shock protein 70 (HSP70) and heat shock protein 90(HSP90) expression, and analysis of liver function biomarkers (AST, ALT). Meanwhile, oxidative stress indicators were measured using biochemical assay kits (GSH, SOD, CAT, MDA, T-AOC), and transmission electron microscopy was employed to observe mitochondrial ultrastructure, thereby assessing the protective effects of SMS on hepatic oxidative stress and mitochondrial damage induced by HS. In vitro, BRL-3A cells were cultured, subjected to HS, and treated with SMS. Cell viability was assessed using the CCK-8 assay, and changes in mitochondrial reactive oxygen species (ROS) levels, mitochondrial permeability transition pore (MPTP) opening, and mitochondrial membrane potential (MMP) were evaluated using fluorescent probes. The results showed that SMS effectively restored HS-induced histopathological damage in rat liver tissues, reduced serum AST and ALT levels, and downregulated the mRNA expression of HSP70 and HSP90 in liver tissues. Meanwhile, SMS strengthened the hepatic antioxidant system by increasing the levels of GSH, SOD, T-AOC, and CAT, while decreasing MDA content. In vitro experiments confirmed that SMS increased the viability of BRL-3A cells, reduced ROS production, improved MPTP opening/closing regulation, and stabilized MMP. This study provides a clinical reference for its application in treating HS-related conditions in humans and animals. Full article

The purpose of this study was to investigate the effects of different feeding-promoting substances added to high plant protein diets on the growth, antioxidant, serum biochemical parameters, immune, and feeding-related genes of Procambarus clarkii. A total of 450 crayfish (3.94 ± 0.03 g) were selected and randomly divided into six groups, with each group consisting of three replicates and 25 crayfish per replicate. The crayfish were fed a basal diet without attractant (control group) and five experimental diets supplemented with 0.4% betaine (BET), 0.4% trimetlylamine oxide (TMAO), 0.4% squid paste (SQU), 0.4% dimethyl-β-propiothetin (DMPT), and 0.4% taurine (TAU). The feeding trial lasted for 6 weeks. The results showed that compared with the control group, the BET, SQU, DMPT, and TAU groups significantly improved in growth performance, weight gain rate, and specific growth rate of crayfish. Compared with the control group, the BET, MTAO, and SQU groups significantly increased hepatopancreas SOD, CAT, and T-AOC. Histological results showed that compared with the control group, all feeding attractant groups could alleviate hepatopancreas tissue damage. Compared with the control group, the TMAO and SQU groups significantly reduced serum GLU content as well as ACP and AKP activities. The results of gene quantitative analysis showed that, compared with the control, TMAO significantly upregulated the expression of tlr, nf-kb, propo, hsp70, and tgf-β, while TAU significantly increased the expression of hsp70, hsp90 and nf-kb genes. Compared with the control group, the expression levels of tor, 4ebp1, and s6k1 in the TMAO group were significantly increased. Compared with the control group, the expression levels of leptin and npy genes in the DMPT group were significantly increased. In summary, the addition of attractants to high plant protein feed has the effects of promoting growth, enhancing antioxidant capacity, improving digestive enzyme activity, alleviating hepatopancreas injury, improving immunity, and promoting feeding. Full article

Background: Pinellia ternata is a major medicinal herb widely utilized in traditional medicine, but is sensitive to high temperature, which often triggers a severe “sprout tumble” phenomenon. Methods: To elucidate the molecular mechanisms of heat tolerance in P. ternata, we screened two contrasting germplasms: the heat-tolerant JBX1 and the heat-sensitive XBX4. In the present study, a combined analysis of physiology, transcriptome, and metabolome was performed on JBX1 and XBX4 under heat stress at 40 °C. Results: JBX1 exhibited significantly greater leaf thickness, higher basal chlorophyll content, more stable antioxidant enzyme activities, and lower oxidative damage than XBX4 under heat stress. Transcriptomically, JBX1 maintained elevated basal expression of genes encoding key enzymes in carbon fixation, amino acid metabolism, and phenylpropanoid biosynthesis, as well as those encoding heat shock transcription factors (HSFs), heat shock proteins (HSPs), and the thermosensor Thermo-With ABA-Response 1 (TWA1). Metabolomically, JBX1 accumulated higher levels of key primary metabolites, antioxidants, and protective phenylpropanoids under both control and heat conditions. Notably, a “polarity reversal” emerged in nitrogen metabolism, where core amino acids accumulated in JBX1 but were depleted in XBX4. Integrated analysis revealed a more coordinated gene–metabolite network in JBX1 involving the phenylpropanoid, ATP-binding cassette (ABC) transporter, and glutathione pathways. Conclusions: Our findings demonstrate that JBX1 possessed stronger basal thermotolerance, which is derived from coordinated establishment of higher constitutive metabolic reserves and efficient dynamic metabolic reprogramming. This study provides insights into the molecular mechanisms of heat stress in P. ternata. Full article

Larix olgensis, a keystone timber species in Northeast China, is increasingly threatened by Neofusicoccum laricinum-induced shoot blight, a devastating disease that compromises forest health and necessitates sustainable management strategies. Here, we demonstrate that the endophytic bacterium Bacillus amyloliquefaciens JL54 elicits multifaceted defense responses in L. olgensis, enhancing resistance to pathogen infection. Greenhouse assays revealed that JL54 pretreatment reduced disease incidence by 12.5% and achieved 43.75% control efficacy while maintaining host vigor. Histochemical analyses identified JL54-induced rapid hydrogen peroxide (H₂O₂) accumulation, extensive lignin deposition, and localized programmed cell death (PCD), indicative of a primed immune response. Transcriptomic analyses uncovered distinct temporal defense patterns: early-stage responses (0 h post-inoculation) were characterized by upregulation of cutin, suberin, and wax biosynthesis pathways, reinforcing physical barriers, whereas late-stage responses (12 h post-inoculation) were dominated by ribosome- and proteostasis-related pathways (e.g., heat shock proteins [HSPs], glutathione S-transferases [GSTs]) to mitigate cellular damage. Biochemical assays corroborated these findings, with JL54 colonization reducing membrane lipid peroxidation (27.2% decrease in malondialdehyde content) and significantly elevating the activity of key defense enzymes, including peroxidase (POD), phenylalanine ammonia-lyase (PAL), and GST. Phytohormone profiling implicated jasmonic acid (JA) as the central mediator of induced systemic resistance (ISR), with JL54-potentiated JA signaling preceding pathogen containment. Collectively, these results demonstrate that JL54 contributes to a coordinated defense strategy in L. olgensis, integrating structural reinforcement (cuticle/lignin), oxidative stress management, and JA-mediated immune priming. These insights advance the understanding of endophyte-conferred resistance in conifers and highlight JL54’s potential as a biocontrol agent for sustainable forestry. Full article

Background/Objectives: The significant negative correlation between protein and oil content in soybean seeds is a long-standing bottleneck for conventional breeding. Its root cause lies in insufficient understanding of related molecular regulatory processes. Methods: We selected the CSSL_R19, a chromosome segment substitution line, to thoroughly investigate the intrinsic effects of the substituted segment on the high seed storage protein (SSP) and low fatty acid (FA) phenotype. Transcriptomic, proteomic, and metabolomic analyses were performed on the recurrent parent and R19. Results: A total of 1821 differentially expressed genes (DEGs), 12 differentially expressed proteins (DEPs), and 10 differentially accumulated metabolites (DEMs) were detected. Subsequently, an integrative examination of the data demonstrated that 28 DEGs, 5 DEPs, and 4 DEMs participated in biological processes such as carbohydrate metabolism, lipid degradation, as well as protein synthesis and transport. Mechanistically, down-regulation of PGM reduces the carbon source supply for FA synthesis; up-regulation of LOX, LACS, ACX, and KAT promotes FA degradation. SRP, SAR1, and HSP70 are involved in the synthesis and transport of SSP. Crucially, qRT-PCR validation performed on all 28 core DEGs showed that their expression trends were highly consistent with the transcriptome data, confirming the reliability of the findings. Conclusions: In conclusion, we propose a potential regulatory network that enhances SSP accumulation and reduces FA content. Altogether, these findings advance our understanding of storage compound accumulation in soybeans and guide future breeding strategies. Full article

Codon usage bias is important for regulating protein translation efficiency and accuracy. The HSP90 gene, a pivotal gene in plants, maintains homeostasis in plant protein stress responses and organelle immune defense functions. We systematically examine codon usage preferences in six forage grass species and the regulatory mechanisms of the HSP90 gene in governing codon preference. A set of metrics is evaluated, including effective codon number (ENC), codon adaptation index, and relative synonymous codon usage. Neutral evolutionary trajectories reveal usage preferences for six plant codons, with natural selection serving as the primary driving factor. The correlation between the ENC–GC3 curve (ENC relative to third-position GC content in synonymous codons) and codon bias index reveals these genes to exhibit moderate codon bias. The phenomenon of evolutionary constraints is exemplified by a propensity for C/G-terminating codons, concomitant with a suppression of NUA/NCG codons (NUA is an abbreviation for UA dinucleotide, and NCG is an abbreviation for CG dinucleotide). Phylogenomic reconstruction reveals a conserved diversification pathway, positioning P. giganteum A. Rich. at the basal node of the evolutionary framework. This study identified through systematic assessment that natural selection is the primary evolutionary force driving the biased use of codons in grass HSP90 genes. This finding provides actionable insights for enhancing abiotic stress tolerance in forage germplasm through precise codon engineering. Full article

Primula vulgaris possesses considerable edible, medicinal, and ornamental value. It is widely applied in food and pharmaceutical development and, as an early-spring flowering plant, is used in landscaping. However, its range of applications and scope are significantly limited due to its inability to withstand high temperatures. This study aimed to investigate the heat tolerance of P. vulgaris under natural high temperatures during summer, identify the most heat-resistant varieties, and determine the optimal conditions for summer outdoor cultivation. Eight P. vulgaris varieties were selected and placed under forest shade with three different shading rates during the summer high-temperature period. Additionally, the heat damage index and the following six physiological indicators were measured: malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, peroxidase (POD) activity, soluble sugar content, soluble protein content, and relative conductivity. Furthermore, a correlation analysis of the physiological indicators was conducted, and a heat tolerance evaluation was performed using the membership function method. Simultaneously, qRT-PCR was employed to analyze the expression patterns of three heat stress-related genes (PvHSP70, PvNCED6, and PvHSF24) across the different cultivars and experimental sites. Under heat stress conditions, leaf area was found to be positively and highly significantly correlated with stomatal density (p < 0.01). The heat damage index, MDA content, and relative conductivity increased significantly with prolonged stress, and they showed highly significant positive correlations. SOD activity, soluble sugar content, and soluble protein content increased to resist heat damage, while POD activity exhibited no consistent trend. Highly significant positive correlations were observed among protective enzyme activities and osmotic regulatory substances. After a comprehensive evaluation, the eight varieties were ranked according to heat tolerance as follows: “Early Punas Yellow” > “Danova Red” > “Middle Punas Rose Red” > “Middle Punas Blue” > “Middle Punas Red” > “Danova Rose White” > “Middle Punas Crimson” > “Middle Punas Scarlet”. Conclusions: “Early Punas Yellow”, “Danova Red”, and “Middle Punas Rose Red” demonstrated strong heat tolerance. In addition, the expression of PvHSP70 and PvHSF24 was significantly upregulated in heat-tolerant cultivars, while that of PvNCED6 showed a sustained increasing trend with rising temperatures. The results of a three-way ANOVA suggested that P. vulgaris exhibited different regulatory patterns among various traits under natural high-temperature stress. Morphological and integrative damage-related indicators, including leaf area, stomatal density, and the heat damage index, all presented significant “site × time” interaction effects. Meanwhile, some physiological regulatory indicators displayed more complex and inconsistent response patterns. These findings further confirm that a dense forest understory grassland is an ideal environment for the summer outdoor cultivation of P. vulgaris. Full article

Soybean seed quality is defined by an inverse relationship between oil and protein content. Understanding the spatiotemporal regulation of this trade-off is crucial for breeding. This study aims to dissect the transcriptomic networks governing carbon and nitrogen partitioning during seed development. Here, transcriptomic and co-expression network analyses were performed on cotyledon and seedcoat tissues of high-protein (HP) and low-protein (LP) soybean cultivars across three seed developmental stages. We identified 4910 HP/LP-specific differentially expressed genes (DEGs), with striking transcriptional alterations in the early developmental stage. Notably, some important DEGs were enriched in carbon/lipid metabolism, protein folding, and hormone/circadian signaling pathways, among which key gene families (e.g., OLEs, SWEETs, HSPs), core regulators (e.g., LACS, L1L, ABF1), and QTL-localized candidate genes (e.g., FA9) were characterized. Mechanistically, C/VIF1-mediated post-translational inhibition of CWINV1 may restrict carbon flux to oil synthesis in HP seeds; upstream circadian/hormone signaling and L1L-sHSPs jointly promote protein deposition, uncoupling the oil–protein trade-off and enabling HP trait formation. In contrast, LP cultivars upregulated SWEETs, OLEs, and LTPs to facilitate high carbon flux into lipid biosynthesis and storage. These findings provide valuable genetic targets for precision breeding programs aimed at optimizing resource allocation. Full article

The escalating consumption of red meat is a potent environmental risk factor for inflammatory bowel disease (IBD), which is characterized by compromised expression of the xenobiotic transporter P-glycoprotein (MDR1/ABCB1). While gut microbiota metabolize dietary tryptophan into diverse indole derivatives that function as aryl hydrocarbon receptor (AhR) ligands, their differential regulation of MDR1 remains an unresolved AhR paradox. Here, we investigated the mechanisms by which two distinct metabolites, indole-3-acetic acid (IAA) and skatole, regulate MDR1 expression in human colonic epithelial Caco-2 cells. We observed that IAA selectively enhances MDR1 protein stability via an AhR-dependent pathway without inducing de novo transcription, suggesting a mechanism we term enhanced proteostasis mediated by the AhR-Hsp90 complex. Conversely, skatole, a toxic dysbiotic metabolite linked to red meat intake, triggered a time-dependent depletion of MDR1 and potently abrogated the protective efficacy of IAA. Our findings are consistent with a model in which skatole acts as a putative structural disruptor, potentially destabilizing the chaperone complex essential for MDR1 integrity. This destruction is facilitated by a key bacterial enzyme, indoleacetate decarboxylase (IAD), which is a pH-dependent metabolic switch in the gut. The modern Western diet, characterized by high protein and low fiber content, elevates colonic pH, thereby activating IAD to convert protective IAA into toxic skatole. These findings provide a molecular framework for the red meat–microbiome–barrier failure axis and highlight the restoration of the IAA/skatole balance through dietary intervention as a promising therapeutic strategy. Full article

Soil salinization poses a major constraint to global agriculture. Apocynum venetum, a salt-tolerant halophyte, provides an effective model for investigating salt-adaptive strategies; however, the temporal dynamics of its tolerance-associated genes and metabolites remain unclear. In this study, integrated transcriptomics, metabolomics (UHPLC-MS), physiological assays, and weighted gene co-expression network analysis (WGCNA) were conducted to characterize early (7-day) and late (18-day) responses to 200 mM NaCl stress. NaCl stress significantly reduced chlorophyll content while increasing Na⁺ accumulation, MDA levels, antioxidant enzyme activities (SOD and CAT), and total flavonoid content. Early responses (NaCl7) were marked by accumulation of ferulic acid, rhamnetin, and 3,4-dihydrocoumarin, with activation of plant hormone (ABA, auxin, zeatin) and MAPK signaling pathways. Late responses (NaCl18) exhibited increased accumulation of scopoletin, formononetin, and caffeyl-alcohol, with enrichment of phenylpropanoid biosynthesis, glutathione metabolism, and photosynthesis-related pathways. WGCNA identified early-response hub genes, including AOC, MAPKKK17/18, CYP98A, and CCoAOMT, coordinating stress signaling and antioxidant metabolism. Late stress responses involved genes like CPK, GST, CYCD3, and ARF, modulating calcium signaling and ROS detoxification. Genes shared across phases included CYP90C1, HD-ZIP, HSP20, and PP2C, regulating protein stabilization and stress signaling. These findings reveal a two-phase salt tolerance strategy in A. venetum, integrating early signaling and late metabolic adaptation. Full article

Cyrtorhinus lividipennis, a key natural enemy of the brown planthopper, Nilaparvata lugens, has been observed to tolerate short-term high-temperature exposure; however, the physiological and molecular mechanisms underlying this heat tolerance remain unclear, which may hinder its effective conservation and utilization. Here, we combined physiological and biochemical assays with transcriptome sequencing to elucidate the physiological and molecular mechanisms of heat tolerance in C. lividipennis following 1 h exposure to three temperatures: 26 °C (control), 33 °C (moderate heat stress), and 40 °C (severe heat stress). At 40 °C, sorbitol, trehalose, lipid, and glycogen contents increased significantly, whereas glycerol levels declined. Transcriptomic profiling revealed temperature-dependent DEGs enriched in starch and sucrose metabolism, galactose metabolism, glycerolipid metabolism, oxidative phosphorylation, and protein folding, sorting, and degradation, with pronounced temperature-dependent upregulation of heat shock protein (HSP) gene families. Together, these results demonstrate that C. lividipennis coordinates its heat stress response through soluble polyol accumulation, which is known to act as a compatible osmolytes that help stabilize proteins and membranes and mitigate thermal damage, energy metabolic reprogramming, and HSP-mediated proteostasis, thereby providing a theoretical basis for its conservation and utilization in sustainable paddy agroecosystems. Full article

Objectives: Contrast-induced acute kidney injury (CIAKI) is a major cause of hospital-acquired renal injury, and strategies for its treatment are currently lacking. This study aimed to investigate the amelioration effect and mechanism of purpurin, a natural antioxidant, against CIAKI via an integrated analysis of network pharmacology, bioinformatics, molecular docking, and animal experiments. Methods: Network pharmacology approaches were used to predict key targets of purpurin against CIAKI. The differential expression of these key targets was further investigated using bioinformatics analysis and molecular binding with purpurin by molecular docking. A CIAKI model was established in SD rats via iohexol administration, and they were treated with 2.5 mg/kg or 5 mg/kg purpurin. Related physiological and pathological indexes were detected to explore the intervention mechanism. Results: Key gene targets were screened from protein–protein interaction networks, of which Pik3c2a, Esr1, Aktip, HSP90AA1, Bcl2, Caspase3, and SRC in the CIAKI group of GSE189881 were significantly differentially expressed compared to the control group. Molecular docking results show that PI3K, ESR1, HSP90, CASP3, AKTI, and SRC had the highest level of connectivity with purpurin. In vivo experiments demonstrated that the Scr and BUN increased in CIAKI rats, the pathological morphology of renal tissue deteriorated, the levels of TNF-α, IL-1β, and IL-6 increased, the contents of MOD and NO in oxidative stress increased, and the activity of SOD and GSH-PX decreased. After administration of purpurin, the above indexes improved in a dose-dependent manner (<0.05). Western blotting showed that purpurin inhibited the Beclin1/Bcl-2/caspase-3 apoptotic cascade and induced the P62/LC3 autophagy pathway. Conclusions: This study provides experimental evidence supporting purpurin as a potential therapeutic agent for CIAKI and further explores its antioxidant mechanisms. Full article

Cadmium (Cd) is a typical pollutant with strong toxicity even at low concentrations. In the marine environment, Cd is a problem of magnitude and ecological significance due to its high toxicity and accumulation in living organisms. The clam Meretrix meretrix is a useful bioindicator species for evaluating heavy-metal stress. This study investigated the extent of recovery from Cd²⁺-induced oxidative and immune impairments in M. meretrix gills achieved by short-term depuration. Clams were exposed to 3 mg/L Cd²⁺ for six days or three days followed by three days of depuration, and the Cd contents, morphological structure, osmoregulation, oxidative stress, and immune responses in the gills were evaluated. The results showed that gill Cd contents increased with exposure, reaching 9.857 ± 0.074 mg·kg⁻¹ on day 3 but decreased slightly to 8.294 ± 0.056 mg·kg⁻¹ after depuration, while reaching 18.665 ± 0.040 mg·kg⁻¹ on day 6 after continuous exposure. Histological lesions, including lamellar fusion, hemolymphatic sinus dilation, and ciliary degeneration, partially recovered after depuration. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels decreased significantly, while DNA-protein crosslinking rate (DPC) and protein carbonyl (PCO) showed minor reductions. Total antioxidant capacity (T-AOC) and the activities of Ca²⁺/Mg²⁺-ATPase (CMA), cytochrome c oxidase (COX), succinate dehydrogenase (SDH), and lactate dehydrogenase (LDH) increased by over 10% during depuration, though these changes were not statistically significant. Lysozyme (LZM) activity and MT transcript levels increased progressively with Cd exposure, indicating their suitability as biomarkers of Cd stress. Acid and alkaline phosphatase (ACP, AKP) activities and Hsp70 and Nrf2 mRNA transcripts exhibited inverted U-shaped response consistent with hormetic response. ACP and AKP activity levels rose by more than 20% after depuration, suggesting partial restoration of immune capacity. Overall, Cd exposure induced oxidative damage, metabolic disruption, and immune suppression in M. meretrix gills, yet short-term depuration allowed partial recovery. These findings enhance understanding of Cd toxicity and reversibility in marine bivalves and reinforce the usage of biochemical and molecular markers for monitoring Cd contamination and assessing depuration efficiency in aquaculture environments. Full article

Edible Grass (EG) is a hybrid vegetable variety valued for its high biomass and protein content, garnering significant interest in recent years for its potential in food, feed, and health product applications. However, in subtropical climates, intense light and high temperatures severely affect the growth and development of Edible Grass (EG), leading to substantial reductions in yield and quality. This study was conducted in the subtropical humid monsoon climate zone of Changsha, Hunan, China, comparing two growth conditions: natural light (CK) and shading treatment (ST). High light-aggravated heat damage under CK significantly reduced EG yield and quality (p < 0.05), with severe cases leading to plant death. and could even lead to plant death in severe cases. Specifically, maximum air and leaf temperatures under CK reached 38.85 °C and 38.14 °C, respectively, well exceeding the plant’s optimal growth range. Shading treatment (ST) effectively alleviated this damage, significantly increasing the net photosynthetic rate, stomatal conductance, and intercellular CO₂ concentration, while decreasing leaf temperature and transpiration rate (p < 0.001). The analysis of physiological and biochemical indicators indicates that after ST, the activities of SOD, CAT, and POD in the leaves decreased, while the contents of MDA and H₂O₂ were significantly lower compared to the CK group (p < 0.001). The transcriptome sequencing results indicate that a total of 8004 DEGs were identified under shading treatment (ST) relative to natural light (CK), with 3197 genes upregulated and 4807 genes downregulated. Significantly enriched Gene Ontology (GO) terms include ‘cell membrane’, ‘extracellular region’, and ‘protein kinase activity’, while significantly enriched KEGG metabolic pathways include ‘plant hormone signal transduction’, ‘photosynthesis–antenna proteins’, and ‘glutathione metabolism’. Compared to CK, the expression of genes associated with oxidative stress (e.g., CAT1, OXR1, APX, GPX) was significantly downregulated in ST, indicating a relief from light-aggravated heat stress. This transcriptional reprogramming was corroborated by metabolomic data, which showed reduced accumulation of key flavonoid compounds, aligning with the downregulation of their biosynthetic genes as well as genes encoding heat shock proteins (e.g., Hsp40, Hsp70, Hsp90). It indicated that plants switch from a ‘ROS stress–high energy defense’ mode to a ‘low oxidative pressure–resource-saving’ mode. Collectively, ST significantly alleviated the physiological damage of forage grasses under heat stress by modulating the processing of endoplasmic reticulum heat stress proteins, plant hormones, and related genes and metabolic pathways, thereby improving photosynthetic efficiency and yield. The findings provide a theoretical basis for optimizing the cultivation management of EG, particularly in subtropical regions, where shade treatment serves as an effective agronomic strategy to significantly enhance the stress resistance and yield of EG. Full article

Exosomes are small extracellular vesicles found in body fluids. They contain proteins and nucleic acids that reflect the condition of the parent cell. Because of this, they may be useful in colorectal cancer (CRC) diagnostics. In this study we compared three plasma isolation methods: ultracentrifugation (UC), size exclusion chromatography (SEC), and a commercial Exo-spin kit. The material obtained with each method was checked for protein content, purity, and vesicle integrity using protein measurements, Western blots, and scanning electron microscopy. From these results SEC was chosen for further use. Exosomes isolated with this method were tested by an enzyme-linked immunosorbent assay (ELISA) for four CRC-related proteins: HSP70, CK19, CA125, and TAG72. Patient samples showed higher levels of HSP70, CK19, and CA125 than controls. TAG72 levels did not differ. In addition, men had more HSP70 than women. These findings show that SEC can be applied for exosome isolation from plasma and that specific proteins detected in exosomes, including HSP70, CK19, and CA125, may serve as promising markers for non-invasive colorectal cancer diagnosis and monitoring. Full article