Vulvovaginal candidiasis (VVC) is a prevalent fungal infection affecting the female reproductive tract. Although conventional therapeutic approaches for VVC are relatively well-established, they still exhibit certain limitations. Pulsatilla decoction, a classic traditional Chinese medicine formula, has demonstrated significant clinical efficacy in the treatment of VVC. However, its precise mechanism of action remains incompletely elucidated. Objective To clarify the therapeutic mechanism of the n-butanol extract of Pulsatilla decoction (BEPD) on VVC through network pharmacology and animal experiments. Methods A mouse model of VVC was established and the therapeutic effect of BEPD on VVC was evaluated. Network pharmacology was employed to screen the potential signaling pathways of BEPD on VVC. Western blotting, immunofluorescence, immunohistochemistry, and real-time fluorescence quantitative PCR were employed to measure the changes in autophagy, apoptosis, and related pathway proteins in the vaginal mucosa of mice. Results Network pharmacology analysis identified PIK3R1 and AKT1 as key targets of Pulsatilla decoction in exerting antifungal activity against VVC. KEGG pathway enrichment analysis indicated that Pulsatilla decoction exerted its therapeutic effects on VVC by regulating the PI3K-Akt signaling pathway. Animal experiments confirmed that compared with the VVC model group, the BEPD treatment down-regulated the expression of PI3K, p-Akt, and p-mTOR, significantly up-regulated the expression of autophagy-related proteins LC3B and ATG5, significantly inhibited the expression of apoptosis-related proteins Bax and Cleaved-Caspase-3, and significantly promoted the expression of anti-apoptosis-related protein Bcl-2. Conclusion BEPD may promote autophagy and inhibit apoptosis of vaginal epithelial cells by inhibiting the PI3K-Akt-mTOR signaling pathway, thereby restoring the homeostasis of the vaginal mucosal epithelial barrier and alleviating VVC.

Objective To construct a recombinant Escherichia coli strain for the expression of the bacteriophage-derived lytic enzyme Lys162, an efficient and broad-spectrum recombinant enzyme, thus providing a technological foundation for developing novel antimicrobial agents. Methods On the basis of the whole-genome sequencing data of bacteriophage pEC.M2929.1AR.1, the protein structure was predicted via bioinformatics tools, and molecular docking analysis was performed to evaluate the substrate-binding affinity. The expression vector pET28a(+)-Lys162 and the engineered E. coli BL21(DE3) expression system were constructed. Lys162 was further assessed for its environmental stability, in vitro antibacterial activity, and lytic spectrum. Results Structural analysis predicted that Lys162 was an N-acetylmuramidase-type lytic enzyme containing a conserved catalytic domain. Molecular docking confirmed its high-affinity binding to peptidoglycan. The enzyme was expressed in a soluble form in E. coli BL21(DE3) and purified to reach a concentration of 1.89 mg/mL. In vitro assays demonstrated that Lys162 at 125 μg/mL exhibited significant lytic activity against E. coli M2929.1AR, along with potent lytic effects against multiple pathogenic bacteria including Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter spp. The enzyme retained stable activity within a pH range of 4.0-11.0 and at temperatures between 4 ℃ and 60 ℃. Conclusion Lys162 transcends the host specificity of its parental phage, demonstrating broad-spectrum antimicrobial activity and considerable environmental adaptability. Its synergistic effect with EDTA suggests a practical strategy for performance optimization. These results establish a foundation for developing novel enzymatic antimicrobials to address challenges associated with bacterial antibiotic resistance.

Objective The immunoinflammatory response induced by spinal cord injury is a key factor hindering the recovery of neurological functions. Recent studies have shown that gut microbiota dysbiosis can participate in the immune regulation of the central nervous system through the gut-spinal cord axis. This study aims to explore whether curcumin can exert its protective effect on spinal cord injury by reshaping the gut microbiota and thereby regulating the local Treg/Th17 balance in the spinal cord. Methods Female Sprague-Dawley rats weighing 200‒220 g were randomly assigned into the sham operation group, spinal cord injury group, curcumin group, fecal microbiota transplantation group, fecal microbiota transplantation+ curcumin group, and fecal microbiota transplantation+curcumin+GPR inhibitor group. Neurological function recovery was evaluated based on the Basso-Beattie-Bresnahan motor function score and gait analysis. Histopathological changes in the injured area were observed via hematoxylin-eosin staining, Nissl staining, and Luxol Fast Blue staining. RT-qPCR, ELISA, and Western blotting were employed to quantify the expression levels of key transcription factor forkhead box protein 3 (FOXP3) for Treg cells, anti-inflammatory cytokines interleukin (IL)-10 and transforming growth factor (TGF)-β1, as well as key transcription factor retinoic acid receptor-related orphan receptor gamma t (RORγt) for Th17 cells and pro-inflammatory cytokines IL-17 and IL-6 in the spinal cord of each group. Results Compared with the spinal cord injury group and fecal microbiota transplantation group, the curcumin group and fecal microbiota transplantation+ curcumin group showed the most significant improvement in neurological function, specifically manifested by significant increases in BBB motor function scores and gait coordination, along with a marked reduction in the scope of spinal cord injury. At the molecular level, the two groups showed significantly upregulated gene and protein levels of FOXP3, IL-10, and TGF-β1 and significantly inhibited expression of RORγt, IL-17A, and IL-6 in the spinal cord tissue. This suggests that after curcumin intervention in the gut microbiota, the immune balance shifted toward a Treg-dominated anti-inflammatory state. Notably, the aforementioned beneficial effects of curcumin-modified gut microbiota were reversed after combined use of the GPR inhibitor. Conclusion This study indicates that curcumin can act on the gut microbiota to promote the recovery of motor function after spinal cord injury. Curcumin may exert the effect by activating the GPR signaling pathway, thereby upregulating Treg viability, inhibiting Th17 differentiation, and ultimately correcting the Treg/Th17 imbalance. This provides new experimental evidence and application value for using curcumin and its modified gut microbiota as an adjuvant therapeutic strategy for spinal cord injury.

Nitrogen deposition is a major driver shaping the structures and functions of forest ecosystems worldwide. When nitrogen inputs exceed ecosystem critical loads (CLs), significant changes in the diversity and abundance of understory herbaceous plants can occur. This study aims to systematically compile and integrate critical load data for understory herbaceous plants in response to nitrogen deposition across three mycorrhizal types: arbuscular mycorrhiza (AM), ectomycorrhiza (ECM), and mixed arbuscular-ectomycorrhizal forests (AM+ECM), in forests. By establishing a dedicated and standardized database, this work facilitates comparisons of herbaceous plant responses to nitrogen inputs among different mycorrhizal types in forests and provides a scientific basis for assessing the impacts of nitrogen deposition on forest microbe-plant systems. On the basis of the published literature and the global nitrogen deposition critical load database developed by Wilkins et al., relevant data were systematically collected, screened, and standardized to construct the Database of Critical Loads of Nitrogen Deposition for Understory Herbaceous Plants across Different Mycorrhizal Types in Forests. All critical load values were consistently derived via the threshold indicator taxa analysis (TITAN) method. A rigorous quality control workflow was applied, including cross-validation of mycorrhizal types, outlier detection and treatment, and data standardization. The database contains 3 592 standardized records. The core data table includes the following fields: Latin name of herbaceous plant species, forest alliance, mycorrhizal types (AM, ECM, or AM+ECM), species-level critical load values (zenv.cp) estimated by TITAN with corresponding bootstrap uncertainty intervals (5th, 10th, 50th, 90th, and 95th percentiles), response direction (increase or decrease in abundance), purity and reliability metrics, community-level change points (CCP), and associated environmental metadata. The database covers the three major mycorrhizal types as well as graminoid and non-graminoid herbaceous functional groups in forests. This database represents the first large-scale, standardized database explicitly focusing on the relationships among mycorrhizal types, understory herbaceous plants, and nitrogen deposition critical loads in forests. Its standardized structure, transparent metadata, and stringent quality control procedures ensure its reliability for future research and applications, including nitrogen deposition risk assessment, comparative analyses of mycorrhizal functions, ecological model parameterization, and the formulation of biodiversity conservation strategies.

Objective With the widespread use and promotion of plastic film mulching on the Qinghai-Xizang Plateau, a series of issues caused by its application have also emerged. Given the fragile eco-environment of the plateau, it is necessary to investigate the effects of different types of plastic film mulching on the soil microbial community structures in farmland ecosystems. Methods Three treatments—pre-planting soil (ZQ), soil covered with conventional polyethylene mulch (CMPs), and soil covered with biodegradable mulch (BMPs)—were established. Soil physicochemical properties were measured, and high-throughput sequencing of the 16S rRNA gene and ITS region was employed to analyze microbial diversity, community structure, and their associations with environmental factors, on the basis of which the impacts of mulch types on soil microorganisms were evaluated. Results Differences in soil physicochemical factors were observed among different treatments (P<0.05). There were no significant differences in alpha diversity indices for both bacteria and fungi among the treatments, indicating that short-term plastic film mulching did not significantly alter the richness and diversity of microbial communities. The dominant bacterial phyla were Pseudomonadota, Actinomycetota, Acidobacteriota, and Chloroflexota, with most dominant genera being unclassified. The dominant fungal phyla were Ascomycota, Basidiomycota, Mortierellomycota, with dominant genera including Mortierella and Solicoccozyma. Network analysis revealed that the main drivers of bacterial and fungal community structures were pH and microplastic (MP) content, respectively. This result reflected functional differences of fungi and bacteria. Fungi, as primary decomposers, were more sensitive to MP pollution, whereas bacterial community structure was more closely related to soil pH. Functional prediction showed that, in bacteria, only the metabolism pathway within the KEGG level 1 showed a positive correlation with the mulching treatment, and no significant differences in COG functions were observed between treatments. In fungi, saprotrophic functions predominated, and their relative abundance changed significantly among treatments. Conclusion Short-term plastic mulching does not significantly affect microbial alpha diversity, but alters the community structure. Compared with conventional PE mulch, biodegradable mulch shows greater potential in enhancing soil nitrogen and organic carbon pools. However, it leads to more severe short-term MP accumulation, accompanied by the risk of pathogenic fungal enrichment. Therefore, its long-term ecological effects require further assessment.

Objective To prepare reference material (RM) for positive serum specific for genotype Ⅱ African swine fever virus (ASFV) for serological detection, quality control, and proficiency testing (PT). Methods The anti-serum collected from specific pathogen-free (SPF) swine immunized with inactivated genotype Ⅱ ASFV was used as raw material for the preparation of RM. Indirect enzyme-linked immunosorbent assay (iELISA) was employed to evaluate the purity, specificity, homogeneity, and stability of RM. In addition, RM was characterized by nine laboratories and applied in clinical trials by three laboratories. Results A total of five hundred bottles of RM for positive serum specific for genotype Ⅱ ASFV strain HLJ/18 were successfully prepared. The results indicated that the RM we prepared was pure, homogenous, and free of exogenous virus contamination, showing good specificity, homogeneity, and stability. The RM was stable for at least 18 months when it was stored at -20 ℃ and for at least 7 days at 4 ℃, 25 ℃, and 37 ℃. The characterization by the nine laboratories showed that the RM was positive for antibodies against genotype Ⅱ ASFV. Conclusion The positive serum specific for genotype Ⅱ ASFV strain HLJ/18 has successfully been prepared, providing critical material for ASF detection and diagnosis.

Porcine enteric coronaviruses (PECs) include porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine deltacoronavirus (PDCoV). Infections with PECs can cause severe diarrhea in pigs, particularly newborn piglets, resulting in high mortality rates and posing a serious threat and economic loss to the global swine industry. Such infections induce oxidative stress to activate various transcription factors and alter their transcriptional pathways, thereby affecting cellular metabolism and the viral life cycle. This leads to cellular dysfunction and further promotes viral replication, forming a vicious cycle. The oxidative stress associated with PECs is considered one of the potential common pathogenic mechanisms. This review summarizes the information about the oxidative stress induced by infections with PECs and emphasizes that antioxidant strategies represent one of the effective approaches to counteract such infections.

The general stress response (GSR) is a global regulatory strategy developed by bacteria to adapt to diverse environmental stresses by coordinating a suite of physiological and metabolic changes, thereby enabling survival in fluctuating conditions. The alternative sigma factor RpoS (σ^S) serves as a central GSR regulator in bacteria and is crucial for bacterial responses to various stress conditions. Such regulators in bacteria are conserved, while polymorphic variations in rpoS are prevalent across numerous natural isolates and acclimated strains. This polymorphism reflects the adaptive trade-off mechanism formed by bacteria during the evolutionary process, positioning RpoS as a key model for investigating fitness trade-offs in bacteria. This review summarizes the functions and polymorphisms of RpoS and explores the potential environmental drivers underlying its polymorphism.

Objective To investigate the function of ring finger protein 31 (RNF31) in the replication of foot-and-mouth disease virus (FMDV) and to provide a theoretical basis for the research on the molecular mechanism by which the host protein RNF31 regulates FMDV replication. Methods CRISPR/Cas9 gene editing was employed to design two sgRNA sequences in the exon segment of RNF31, and recombinant plasmids were constructed by ligation with the pX459-puro vector. The recombinant plasmids pX459-RNF31-sgRNA were transfected into PK-15 cells, followed by screening under the action of puromycin to obtain the cell lines with RNF31 gene knockout. The effect of RNF31 gene knockout on FMDV replication was detected by Western blotting, RT-qPCR, and TCID₅₀ methods. Results Compared with wild-type cells, the knockout of RNF31 significantly increased the protein level, mRNA level, and virus titer of FMDV. Conclusion We successfully construct the cell lines with RNF31 gene knockout and prove that RNF31 plays a key role in the replication of FMDV. This result provides data support for further research on the mechanism by which RNF31 inhibits FMDV replication.

The gut microbiota plays a crucial role in promoting food digestion in animals. However, the impact of cross-species microbiota transplantation from donors with different dietary habits on the host food digestion capacity remains unclear. Objective To investigate the role of cross-species microbiota transplantation in regulating the digestive system adaptability, metabolic functions, reproduction, stress responses, and gut microbiota structure of the host. Methods We utilized New Zealand white rabbits (Oryctolagus cuniculus), a herbivorous species, and C57BL/6J mice, an omnivorous species, as donors and recipients of gut microbiota, respectively. The mice were allocated into three groups: a control group on a normal diet (Con), a group on a high-fiber diet (TS), and a group on a high-fiber diet supplemented with rabbit fecal microbiota transplantation (OC). This study was designed to evaluate various physiological and biochemical parameters, including body weight, food intake, absolute and relative organ weights (both wet weight and organ-to-body weight ratio), morphometric indices (length and diameter) of the small intestine, sperm concentration, and serum corticosterone level, in mice. Additionally, we performed 16S rRNA gene sequencing targeting the V3-V4 hypervariable region to characterize the composition of fecal microbiota. Results A high-fiber diet significantly increased the food intake, small intestine length, and serum corticosterone level, while significantly reducing the body weight, liver and spleen wet weights, liver/body weight ratio, spleen/body weight ratio, and sperm concentration in mice. Moreover, it increased the alpha diversity of the gut microbiota, decreased the Bacillota-to-Bacteroidota ratio, and reduced the relative abundance of probiotics (such as Ligilactobacillus). Transplantation of the gut microbiota from rabbits increased the wet weight of the epididymis and the epididymis/body weight ratio, while significantly reducing the liver/body weight ratio and the serum corticosterone level in recipient mice. Furthermore, a high-fiber diet significantly increased the relative abundance of the fiber-degrading bacterial family (Oscillospiraceae) and the gut health-associated bacterial genus (Colidextribacter). After the transplantation of rabbit gut microbiota into mice, the relative abundance of Oscillospiraceae and Colidextribacter in mice increased significantly. Conclusion The high-fiber diet has adverse effects on omnivores. Although the microbiota transplantation from herbivores does not significantly improve the host ability to digest fiber, it changes the gut microbiota structure of omnivores, playing a positive role in improving their digestion, reproduction, metabolism, and stress responses. Future research needs to further determine the optimal levels of dietary fiber for omnivores and the dosage of microbiota transplantation from herbivores, as well as their synergistic effects and underlying mechanisms in improving animal health. This study provides a reference for exploring the role of gut microbiota in animal adaptation to dietary changes in natural environments and lays a foundation for future research on improving the utilization of high-fiber foods by omnivorous domestic animals.