Search Results (5,843)

Oxidative stress is one of the few defined causes of male infertility affecting at least one third of patients attending infertility clinics. Human spermatozoa are vulnerable to this form of attack because their stripped-down architecture means that they possess limited antioxidant protection and little capacity for biochemical repair. They also compound their vulnerability by being active generators of reactive oxygen species (ROS) and possessing multiple substrates for oxidative damage. The major sources of ROS in these cells are their mitochondria, an L-amino acid oxidase (IL4I1) and a calcium-dependent NADPH oxidase (NOX5). Spermatozoa tolerate the risks associated with ROS generation because their biology is heavily dependent on redox regulation. ROS are important mediators of sperm capacitation, stimulating the generation of cAMP and prostaglandins, inhibiting protein phosphatases and encouraging removal of cholesterol from the plasma membrane. Furthermore, during fertilization, the ability of ROS to activate metalloproteinases facilitates penetration of the zona pellucida and sperm–oocyte fusion. While ROS are physiologically important for sperm function, the over-production of these metabolites can impair sperm function. Antioxidants have therefore assumed some importance as a possible therapy for the infertile male. However, before this potential can be realized, we need to optimize the composition and dose of reagents used in such formulations and develop improved methods of diagnosing oxidative stress within the patient population. Full article

Objectives: Pilose antler protein extract (PAE) was investigated for its therapeutic efficacy against ovariectomy (OVX)-induced osteoporosis and its underlying molecular mechanism. Methods: An OVX rat model was established to evaluate the effects of PAE on bone microarchitecture, histopathological changes, and bone metabolism-related parameters. Bone structure was assessed using Micro-CT and histological analysis, and biochemical and bone turnover markers were quantified. In vitro, Mouse calvarial pre-osteoblast subclone 14 (MC3T3-E1) Subclone 14 cells were used to examine the effects of PAE on cell viability, proliferation, osteogenic differentiation, and osteogenesis-related protein expression. Results: High-dose PAE markedly improved trabecular bone microarchitecture in OVX rats, as reflected by increased bone surface area and bone volume fraction and reduced trabecular separation. PAE significantly enhanced bone calcium content and elevated serum Bone Morphogenetic Protein 2 (BMP-2) and Procollagen Type I N-Terminal Propeptide (PINP) levels, while decreasing serum Alkaline Phosphatase (ALP) activity and C-Terminal Telopeptide of Type I Collagen (CTX-I) levels, indicating a shift toward bone formation. Mechanistically, PAE activated the Wnt-3a/β-Catenin signaling pathway in bone tissue and MC3T3-E1 cells, as evidenced by increased expression of Wnt-3a and β-Catenin proteins. In vitro experiments further demonstrated that PAE promoted MC3T3-E1 cell proliferation and upregulated osteogenic markers, i.e., Runt-related transcription factor 2 (RUNX2) and Osteocalcin (OCN). Conclusions: Collectively, these findings suggest that PAE exerts pronounced anti-osteoporotic effects and is associated with activation of the Wnt/β-Catenin signaling pathway. Full article

This study evaluated whether three-dimensional alginate–gelatin hydrogels (AGHs) crosslinked with calcium chloride (CaCl₂) enhance the osteo-odontogenic differentiation of odontoblast-like cells in vitro. Two seeding configurations were compared: inter-hydrogel (INT) surface seeding and intra-hydrogel (INTR) encapsulation. Here, the MDPC-23 cells were cultured in AGHs crosslinked with 70 or 100 mM CaCl₂ and assessed for proliferation, cytoskeletal morphology, alkaline phosphatase (ALPase) activity, osteo-odontogenic gene expression, and mineralized nodule formation. After 7 days, cell proliferation was significantly greater in the alginate–gelatin hydrogel (AGH) groups than in the control group. Cells in the intra alginate–gelatin hydrogel 100 (INTR-AGH100) remained predominantly rounded, whereas those in the inter alginate–gelatin hydrogel 100 (INT-AGH100) formed irregular clusters on the hydrogel surface. ALPase activity was highest in INTR-AGH100 at the early stage of culture. Both INT-AGH100 and INTR-AGH100 showed significantly increased expression of DSPP, DMP-1, BSP, OCN, OPN, and Runx-2, together with enhanced mineralized nodule formation. Although no significant differences were detected between the two seeding strategies in all assays, distinct morphological patterns were observed, and the INTR configuration showed relatively greater early differentiation-related activity. These findings suggest that 100 mM CaCl₂-crosslinked AGHs provide a favorable three-dimensional microenvironment under the present experimental conditions and represent a promising in vitro scaffold platform to support future studies of scaffold-guided dentin regeneration. Full article

Phosphorus (P) limitation is prevalent in terrestrial ecosystems. Plants can improve soil P availability through the exudation of organic acids and symbiotic interactions with microorganisms. However, associations between different plant functional groups and phosphorus cycling in P limited karst ecosystems remain poorly understood. To investigate this, the exudation rates of oxalic, citric and acetic acids from fine roots, the contents of carbon, nitrogen, and P in leaves and fine roots, and the contents of oxalic, citric and acetic acids, total P, available P (AP), and microbial biomass P in rhizosphere soils were measured across different plant functional groups in a karst ecosystem in southwestern China. Additionally, the activities of acid and alkaline phosphatases were also analyzed, as well as the relative abundance, community structure, diversity, and co-occurrence network patterns of arbuscular mycorrhizal fungi (AMF) and alkaline phosphatase-encoding (phoD) gene-harboring bacteria. The results showed that both the exudation rates and the contents of organic acids and AP were highest in the tree group, followed by the shrub and grass groups. The AP content of the legume group was significantly higher than that of the non-legume group. The exudation rates of oxalic acid were significantly greater than those of citric and acetic acids. AMF diversities were highest in the shrub and legume groups. The diversities of phoD-harboring bacteria decreased from the tree group to the shrub group and then to the grass group, yet there were no significant differences between the legume and non-legume groups. The communities of both AMF and phoD-harboring bacteria exhibited significant differences among these plant functional groups. The prevalent genera of phoD-harboring bacteria across all groups were Pseudomonas and Halomonas, with Halomonas being particularly prevalent in the legume group. The AMF community was dominated by Glomus, which attained its highest relative abundance in the tree and legume groups. Furthermore, the increased exudation rate and content of oxalic acid were associated with higher relative abundances of Glomus in AMF and Pseudomonas and Bacillus among phoD-harboring bacteria. Structural Equation Model (SEM) analysis demonstrated that plant-exuded organic acids, especially oxalic acid, were positively associated with P availability indirectly through their linkages with the diversity and abundance of AMF and phoD-harboring bacteria. The crucial role of oxalic acid was particularly prominent in the tree and legume groups. Our findings suggest that screening AMF and phoD-harboring bacteria with highly efficient P transformation activity and inoculating them into the rhizosphere of plants with high oxalic acid exudation could help improve plant resilience to P limitation and support sustainable restoration in karst ecosystems. Full article

Purpose: Black soils in Northeast China are declining in fertility under intensive fertilization, motivating strategies that integrate crop residue return with microbial inoculation. We conducted a field experiment to test whether maize straw return combined with compound microbial inoculants improves soil properties, bacterial communities, and soybean performance. Methods: A field experiment compared four treatments: fertilization alone (F), fertilization + inoculants (CF), fertilization and straw (SF), and fertilization and straw with inoculants (CSF). Soil physicochemical properties, enzyme activities, 16S rDNA-based bacterial communities, and soybean agronomic yield were measured across growth stages. Results: CSF produced the highest soybean performance, and increased yield by 3.91–5.46% compared with F. CSF increased soil pH, moisture, and nutrient availability (notably available P and K) and enhanced sucrase, urease, catalase, and acid phosphatase activities compared with other treatments. Bacterial communities were dominated by Acidobacteriota and Proteobacteria. CSF increased bacterial abundance and shifted community composition, and pH and available P were key factors associated with community variation. Conclusions: Co-applying maize straw and compound microbial inoculants enhances soybean yield while improving soil biochemical functioning and reshaping bacterial communities in black soil. Full article

Protected cultivation of pepper in southern China’s red soil region often leads to soil degradation and continuous cropping obstacles. To investigate whether biochar can alleviate these problems by regulating the soil microenvironment, pot and incubation experiments were conducted from 2021 to 2023 with biochar application rates of 0~10% (w/w). The results showed that appropriate biochar application significantly improved pepper yield and soil quality. Under the 6% biochar treatment, pepper yield and dry matter accumulation increased by 89.05% and 36.79%, respectively, compared to the control. Soil bacterial and fungal abundances increased by 346.61% and 107.37%, and their OTU numbers rose by 64.13% and 35.15%, respectively. Biochar application also elevated soil pH, organic matter, available potassium, and total nitrogen contents, improved aggregate stability, and enhanced the activities of urease, catalase, sucrase, and acid phosphatase. Furthermore, biochar altered the rhizosphere microbial community structure and increased bacterial diversity. These findings demonstrate that biochar can promote pepper growth by improving soil physicochemical properties, enzyme activities, and microbial community structure, providing a viable strategy for mitigating continuous cropping obstacles in protected cultivation. Full article

Ubiquitin-specific protease 13 (USP13) is a deubiquitinating enzyme that stabilizes phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a well-established tumor suppressor involved in PI3K/AKT signaling. This study aimed to evaluate the relationship between USP13 immunohistochemical staining intensity and clinicopathological factors associated with prostate cancer progression. USP13 staining was scored as grade 0 (negative), 1 (weak), 2 (moderate), or 3 (strong) in 242 prostate cancer tissues and 22 adjacent non-neoplastic control tissues. Higher USP13 grades were exhibited by adjacent non-neoplastic tissues than prostate carcinoma. In comparison, lower USP13 grades were observed in 88.6% of the neoplastic regions (p < 0.001). No differences in PSA level, Gleason’s score, disease stage, involvement of either the seminal vesicle or lymph nodes, surgical margin positivity, biochemical or clinical recurrence rates, or overall survival statistics were found. Cox proportional hazards modeling showed no significant association between USP13 expression and biochemical recurrence-free survival or overall survival. Kaplan–Meier analysis demonstrated no statistically significant differences in survival outcomes according to USP13 expression, although a descriptive trend was observed. USP13 immunohistochemical staining distinguished malignant prostate tissue from adjacent non-neoplastic tissue in tissue microarrays. However, USP13 expression was not independently associated with pathological aggressiveness or survival outcomes in this cohort. Full article

Soil health is critical for the sustainability of tropical plantation ecosystems, However, the ecological factors driving productivity gradients remain inadequately understood. This study investigated rubber plantations on Hainan Island with varying yield levels to assess soil health and its underlying ecological mechanisms using a minimum data set (MDS) approach. Twenty-seven soil physical, chemical, and biological indicators were analyzed at two depths (0–20 cm and 20–40 cm). Principal component analysis identified seven key indicators for the MDS: soil organic matter (OM), alkaline-hydrolyzable nitrogen (AN), cation exchange capacity (CEC), dissolved organic carbon (DOC), microbial biomass phosphorus (MBP), acid phosphatase activity (ACP), and microbial diversity (Shannon-Wiener index, SHDI). The soil health indices derived from the MDS showed strong correlations with those generated from the total data set (TDS) (p < 0.001), confirming the reliability of the MDS framework. Overall, soil health levels were rated low to moderate with no significant differences across low-yield plantations (≤900 kg·ha⁻¹), medium-yield plantations (900–1200 kg·ha⁻¹), and high-yield plantations (≥1200 kg·ha⁻¹)., suggesting a decoupling of soil health and rubber productivity under uniform management practices. Random forest analysis identified microbial-driven phosphorus cycling, particularly MBP and ACP, as the primary determinant of soil health across soil layers, with DOC and SHDI also contributing significantly. These findings highlight the critical role of microbial-mediated nutrient cycling in maintaining soil health in rubber plantations and suggest that current management practices prioritize short-term yields over long-term soil ecological stability. Enhancing microbial activity and increasing organic matter inputs may be essential for improving soil health and ensuring the sustainability of rubber production in tropical agroecosystems. Full article

Background/Objectives: It is challenging to develop efficient vaccines against intracellular pathogens such as viruses, and since viral infections are one of the main challenges for farmed salmon, a novel vaccine strategy is needed. mRNA vaccines are optimized and approved for humans, but for fish, the mRNA technology is new, and optimization is required to ensure efficient protein expression. We made an mRNA tailored to salmon and studied the effect of modified nucleosides and the length of the poly(A) tail on protein expression from in vitro-transcribed mRNA in CHSE-214 cells, using enhanced green fluorescent protein (EGFP) as a reporter. Methods: Different lengths of the poly(A) tail were tested, and various modified nucleotides were incorporated in the mRNA during in vitro transcription, including pseudouridine (Ψ), N1-methylpseudouridine (m1Ψ), N6-methyladenosine (m6A), 5-methyluridine (m5U), and 5-methylcytidine (m5C). Protein expression was observed in fluorescence microscopy and quantified using flow cytometry. Results: mRNA containing Ψ resulted in the strongest EGFP expression 1–3 days post-transfection (dpt), while EGFP expression from m5C mRNA was high throughout the experiment (<10 dpt). m5U-containing mRNA had low EGFP expression until 6 dpt, but reached the level of m5C mRNA at 10 dpt. The m5U mRNA, however, expressed EGFP at much higher intensity than all the other mRNAs at all time points. Poly(A) tails with lengths of 40, 100, and >100 were tested, and the one with >100 adenines showed the highest expression. The effects of phosphatase treatment and purification of the mRNA were also investigated. Furthermore, EGFP expression was observed in yolk-sac salmon larvae following micro-injection. Conclusions: Our study provides an important basis for the development of efficient mRNA-based vaccines in the future. Full article

Ledum palustre (L. palustre) is widely used in drug development because of its antibacterial and analgesic effects. However, wild L. palustre is often affected by wildfires, resulting in unstable yields. Forest fires represent a major disturbance in northern forest ecosystems and profoundly affect shrub vegetation and its associated rhizosphere microbial communities. In this study, we investigated a fire chronosequence (1991, 2004, 2012, 2017, and 2020) to systematically examine the morphological traits of L. palustre, rhizosphere soil physicochemical properties, and microbial community characteristics and to identify the key drivers underlying these patterns. The results revealed that postfire recovery time significantly influenced the morphological traits of L. palustre. The biomass, branch number, basal diameter, and plant height of the shrubs at the 1991 burned site increased by 270.49%, 36.11%, 79.32%, and 191.36%, respectively (p < 0.05). From unburned soils, 29 bacterial and 29 fungal isolates were obtained, with Bacillus sp. and Oidiodendron sp. being the dominant culturable bacterial and fungal taxa, respectively. With increasing postfire recovery time, soil moisture, total nitrogen, ammonium, nitrate, soil organic carbon, acid phosphatase (AP) and N-acetyl-β-D-glucosaminidase (NAG) activity significantly decreased. Early fire disturbance markedly altered soil microbial abundance and community composition, leading to an overall decrease in bacterial α diversity. The bacterial community structure at the 2020 burn site and the fungal community structure at the 2012 burn site significantly differed. Mantel tests revealed significant positive correlations between branch number and basal diameter (p < 0.01) and significant negative correlations between plant height and stem density (p < 0.001). Soil carbon and hydrolysable nitrogen were significantly positively correlated with AP and NAG activities (p < 0.001). Moreover, soil physicochemical properties significantly shaped soil microbial community structures, with bacterial communities in early postfire sites driven by total carbon and nitrogen (p < 0.05), whereas fungal communities in the 2012 burned site were influenced primarily by β-N-acetylglucosaminidase (BG) activity (p < 0.05). Fire disturbance drives successional changes in the rhizosphere microbial community structure and function by altering the soil nutrient status and enzyme activity, which in turn influences the morphological traits of L. palustre. This study provides a theoretical basis for improving the yield of L. palustre by exploring the variation in rhizosphere microorganisms. Full article

Continuous greenhouse watermelon cultivation is widely constrained by declining soil function, impaired nutrient cycling, and increasing soil-borne disease pressure. Developing biologically driven strategies to restore soil–crop coupling is therefore critical for sustainable protected horticulture. Here, we conducted a two-year field experiment (2024–2025) using a randomized block design with three treatments (CK, ST, and STE), three replicates per treatment, and a plot area of 22.5 m² to evaluate how straw application alone and in combination with earthworms regulate soil processes and crop performance in a continuous greenhouse watermelon system. Compared with CK and ST, earthworm–straw co-application (STE) exerted stronger effects, particularly during the mid-to-late growth stages. In 2024, STE increased soil organic matter by 25.34% and 30.28% relative to CK at the fruiting and harvest stages, respectively; in 2025, the corresponding increases were 25.22% and 27.62%. STE also significantly increased total nitrogen at nearly all growth stages, with the maximum increase reaching 67.23% relative to CK at harvest. In 2025, total phosphorus under STE was significantly higher than under CK and ST across all growth stages, with increases of 75.82% and 79.63%, respectively, at the fruiting stage. Neutral phosphatase activity was markedly enhanced, increasing by 292.24% at the fruiting stage in 2025. These improvements were accompanied by higher plot yield and lower wilt disease incidence, with yield increasing by 34.00% in 2024 and 21.29% in 2025 relative to CK, while disease incidence decreased by 41.46% and 56.06%, respectively. Integrative Mantel tests showed that total nitrogen was the factor most strongly associated with watermelon yield, with the correlation coefficient increasing from r = 0.490 (p = 0.001) in 2024 to r = 0.662 (p = 0.001) in 2025. Co-occurrence network analysis further revealed a strong positive correlation between yield and total nitrogen (r = 0.848 in 2024; r = 0.673 in 2025) and a negative correlation between disease incidence and total nitrogen (r = −0.661 in 2024; r = −0.822 in 2025), indicating progressively strengthened soil–plant functional coupling over time. Our findings demonstrate that earthworm–straw co-application strengthened soil nutrient transformation capacity and enhanced soil suppressiveness against wilt disease, thereby providing an effective ecology-based strategy for alleviating continuous-cropping constraints in greenhouse watermelon systems. Full article

Soil nutrient loss and infertility in rocky desertification areas severely constrain ecological restoration. Exploring the impacts of external field remediation technologies on soil quality in these regions may offer novel strategies for soil enhancement and ecosystem recovery. This study conducted a three-month experiment to investigate the impact of continuous electric (ET, 20 V) and magnetic (MT, 200 mT) field treatments on soil nutrients, enzyme activities, and bacterial communities in simulated moderate and severe rocky desertification soils. Results showed that although an overall declining trends in total contents of key soil nutrients (Total nitrogen, total phosphorus, and total potassium), both electric and magnetic field treatments effectively mitigated the decreases of total nitrogen and potassium content (with the exception of total phosphorus) in rocky desertification soils, while improving their available contents compared to the control (CK). Electric field application significantly reduced the pH of moderate and severe rocky desertification soils through electrolysis, shifting the soil from alkaline (pH 7.69 and 7.73, respectively) to slightly acidic (pH 6.71 and 6.37, respectively); Both electric and magnetic field treatments enhanced urease and sucrase activities in moderately and severely rocky desertified soils. Compared to the CK, the increases were 21.92%, 4.46%, 5.70%, and 66.43% in moderately rocky desertified soil, and 10.06%, 42.15%, 20.66%, and 0.93% in severely rocky desertified soil, respectively. Their effects on phosphatase and catalase activities varied with the degree of rocky desertification. However, in severely rocky desertified soil, both treatments significantly increased phosphatase and catalase activities by 19.55%, 24.63%, 61.07%, and 38.05% compared to the CK, respectively. Furthermore, both electric and magnetic treatments significantly reduced bacterial α-diversity (chao1, ACE, Shannon, Simpson, and Pielou J indices) but optimized community structure by enriching dominant phyla with specific ecological functions, such as Pseudomonadota (7.63–41.10%), Bacteroidota (13.52–69.29%), and Verrucomicrobiota (38.26–104.81%). Functional prediction revealed that the abundances of dominant pathways (such as chemoheterotrophy, aerobic chemoheterotrophy, and nitrogen fixation) was enhanced following both treatments. Mantel analysis further indicated strengthened correlations among soil nutrients, enzyme activities, and bacterial communities, particularly under magnetic field treatment. These findings demonstrate that electric and magnetic field applications effectively facilitate nutrient cycling, stimulate enzyme activities, and optimize microbial community structure, thereby improving soil ecological functions and overall quality in rocky desertification regions, highlighting their potential for ecological restoration in karst areas. Full article

(1) Different classes of antidepressant drugs have been shown to activate lysophosphatidic acid (LPA) receptors, but their effects on the receptor signaling stimulated by LPA have not been fully investigated. In the present study, we examined the effect of the tricyclic antidepressant amitriptyline on the LPA-induced activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and Rho signaling in C6 glioma cells and cultured rat astrocytes. (2) LPA receptor signaling was investigated by using Western blot and microscopic immunofluorescence assays. Rho activation was determined by a pull-down assay. (3) Amitriptyline potentiated the LPA-induced activation of ERK1/2 signaling, as indicated by the more than additive increases in the phosphorylation/activation of key components of this pathway including fibroblast growth factor 1 receptor, MEK1/2, ERK1/2, Elk-1, and cyclic AMP response element binding protein (CREB). Amitriptyline also enhanced the expression of brain-derived neurotrophic factor (BDNF) elicited by LPA. In contrast, the antidepressant failed to mimic the LPA-induced activation of Rho and Rho-dependent responses, such as the reversal of astrocyte stellation, accumulation of stress fibers, and the phosphorylation of focal adhesion kinase and myosin target subunit of myosin phosphatase isoform 1. Moreover, when combined with LPA, amitriptyline curtailed Rho activation and the Rho-mediated cellular responses. (4) These results demonstrate that in astroglial cells, amitriptyline exerts a balanced action on LPA-activated receptors by enhancing the neuroprotective ERK1/2-CREB-BDNF signaling and dampening the potentially detrimental Rho–ROCK pathway, and suggest that this unique property may contribute to the antidepressant activity of the drug. Full article

Quantitative detection of alkaline phosphatase (ALP) activity is crucial in clinical diagnosis and bioanalysis. Herein, we have developed a highly sensitive electrochemiluminescence (ECL) biosensor that employs a biomimetic zirconia interface as its core sensing platform. The interface was constructed by immobilizing o-phosphorylethanolamine (PEA) onto zirconium oxide nanofilms (ZrO₂NFs), forming a surface rich in Zr-O-P bonds. This design mimics phosphate recognition and enzyme-triggered dephosphorylation processes, where ALP catalyzes the hydrolysis of these bonds, triggering a direct switch in the ECL signal from Ru(bpy)₃²⁺-loaded gold nanocage (Ru-AuNCs) emitters. This sensor achieves a wide linear range of 0.100–100 U/L and a low detection limit down to 0.0899 U/L. Its practical utility was validated through the accurate detection of ALP in fetal bovine serum samples, confirming high recovery and reliability. This strategy highlights the potential of biomimetic zirconia interfaces in developing robust biosensors for early disease diagnosis. Full article