Hunan Academy of Traditional Chinese Medicine

Deoxynivalenol (DON), a Fusarium-derived mycotoxin widely found in grain-based feed, has become a major global environmental contaminant. Reproductive toxicity is one of its most important toxic effects, yet systematic investigations covering both male and female reproductive injury remain limited. This study aimed to establish a combined strategy of network toxicology, molecular docking, molecular dynamics simulation, and single-cell RNA sequencing to evaluate the reproductive toxicity of DON. AKT1, EGFR, PIK3CA, PIK3R1, and SRC were identified as key targets involved in DON-induced reproductive injury. For testicular injury, the prolactin, Ras, HIF-1, and AGE-RAGE signaling pathways were closely associated with DON toxicity. For ovarian injury, the PI3K-Akt, HIF-1, prolactin, insulin, and AGE-RAGE signaling pathways were strongly implicated. Molecular docking demonstrated favorable binding affinities between DON and the hub targets, while molecular dynamics simulation further confirmed the stability of the DON-PIK3CA complex. Single-cell RNA sequencing analysis revealed that these five hub genes were highly expressed in both testicular (SRA667709:SRS3065430) and ovarian (SRA638923:SRS2797100) tissues. These findings deepen current understanding of DON-induced reproductive toxicity, provide new insights into the effects of environmental toxins on reproductive health, and offer a theoretical basis for future studies integrating DON exposure with in vivo validation of core targets and signaling pathways.

Fecal microbiota transplantation (FMT) is an emerging therapy that has received significant attention in recent years, although its origins can be traced back to 4th-century China. In modern medicine, FMT has been incorporated into clinical guidelines for the treatment of recurrent Clostridioides difficile infection. By re-establishing a healthy gut microbiota and regulating the immune system, FMT has potential therapeutic effects on various diseases, such as gastrointestinal diseases, diabetes, tumors, Alzheimer's disease, and liver disease. However, its efficacy varies based on the type of disease and individual differences. The clinical application of FMT is influenced by multiple factors, including fecal matter processing, administration route, dosage, donor screening, and recipient detection. Currently, FMT faces numerous challenges, including the need to verify the stability and durability of its efficacy, standardize donor screening criteria, and optimize fecal processing and administration. Future research is expected to reveal the mechanisms of action of FMT, optimize treatment protocols, and refine its safety, efficacy, and convenience, thereby bringing hope for patients with complex and challenging diseases.

Ophiocordyceps xuefengensis is an important medicinal fungus with considerable pharmaceutical and economic value. However, its industrial and scientific utilization has been severely limited by the lack of an efficient genetic transformation system, largely due to limited genomic information and wild growth. In this study, we established an efficient and stable plasmid transformation system within O. xuefengensis protoplasts mediated by PEG. To overcome low protoplast yield and transformation efficiency, key factors influencing protoplast preparation including enzyme composition and concentration, fungal age, and digestion conditions were systematically optimized. The optimal protocol involved digesting 4-day-old mycelia with a mixture of 1.5% lywallzyme 1 and 1.5% snailase at 34 °C and 130 rpm for 3.5 h, yielding at least 9.42 × 107 CFU/mL protoplasts. Protoplast regeneration was significantly enhanced in PY medium supplemented with 0.6 M mannitol. Under these optimized conditions, a transformation efficiency of 45.5% was achieved, with stable plasmid integration confirmed over four successive generations. Furthermore, the transformation system was successfully applied to functional gene characterization by driving exogenous gene expression using the endogenous gpd1 promoter. This study provides a foundational platform for functional gene analysis and paves the way for further elucidation of growth and development mechanisms and metabolic engineering in O. xuefengensis.

Recent studies have shown that heat shock protein 90 alpha family class A member 1 (HSP90AA1) interacts with various tumor-associated proteins, regulates their biological activity and stability, and plays an important role in various tumors. However, the role of HSP90AA1 in clear cell renal cell carcinoma (ccRCC) remains unclear. In the study, GEO and TCGA-KIRC databases were used to analyze the expression pattern and clinical significance of HSP90AA1 in ccRCC; immunohistochemistry and Western blot were used to validate HSP90AA1 expression in ccRCC tissues and cell lines; colony formation assays, EdU and TUNEL methods, cell migration and invasion experiments, and a mouse renal orthotopic xenograft tumor model were used to detect the effects of HSP90AA1 overexpression on the biological function of ccRCC; Co-IP and RNA-seq experiments were utilized to explore the downstream regulatory mechanism of HSP90AA1. Our results showed that HSP90AA1 expression was significantly downregulated in ccRCC, and its reduced expression was associated with tumor metastasis. HSP90AA1 overexpression markedly inhibited the proliferation and metastasis ability of ccRCC cells. HSP90AA1 bound to F-box only protein 7 (FBXO7) and accelerated its protein expression. FBXO7 was expressed at low level in ccRCC, and its decreased expression was closely related to unfavorable pathological features of tumors and poor patient prognosis. FBXO7 overexpression promoted cell adhesion molecule 1 (CADM1) expression and suppressed the PI3K-AKT signaling pathway. Knocking down FBXO7 expression on the basis of HSP90AA1 overexpression significantly reversed the cell phenotype inhibition caused by HSP90AA1 overexpression, downregulated CADM1 expression, and activated the PI3K-AKT signaling pathway. In summary, HSP90AA1 exhibited a low expression pattern in ccRCC, and HSP90AA1 overexpression promoted CADM1 expression and inhibited the PI3K-AKT pathway, thereby suppressing the proliferation and metastasis of ccRCC.

To investigate the influence of lactoferrin (LTF) on the anoikis-resistance of nasopharyngeal carcinoma (NPC) cells and explore its relationship with the protein kinase B (AKT) signaling pathway. Anoikis-resistant HNE-1 and HONE-1 NPC cell lines were established. The proliferation and survival of cells were detected by Cell Counting Kit-8 (CCK8), while cell cycle and apoptosis were measured by flow cytometry. The transwell assay was used to evaluate invasion and metastasis abilities. The expression of matrix metallopeptidase-9 (MMP-9), vascular endothelial growth factor-A (VEGF-A), E-cadherin and Vimentin were assessed by Western blot. LTF plasmids and LTF shRNA plasmids were transfected into HNE-1 and HONE-1 cells, respectively, and the expression of E-cadherin, Vimentin, AKT and tropomyosin receptor kinase B (TrkB) proteins was detected by Western blot to clarify the role of LTF in anoikis-resistant NPC cells. Anoikis-resistant HNE-1 and HONE-1 NPC cell lines were successfully established. Compared to parental cells, these anoikis-resistant cells exhibited enhanced survival, reduced apoptosis, and significantly increased invasive ability. They also demonstrated elevated expression of VEGF-A, MMP-9, and Vimentin, alongside decreased E-cadherin, indicating epithelial-mesenchymal transition (EMT). Furthermore, the expression of AKT and TrkB was significantly upregulated in anoikis-resistant cells. Critically, LTF overexpression reversed this aggressive phenotype: it suppressed cell survival and invasion, induced G0/G1 cell cycle arrest, promoted apoptosis, and downregulated the expression of AKT and TrkB. Conversely, LTF knockdown produced opposing effects. Our study revealed that LTF inhibits anoikis-resistance and metastasis of NPC cells via the AKT signaling pathway.

Sepsis and atrial fibrillation are complex, life-threatening medical conditions affecting approximately 49 million individuals globally, characterized by exceptionally high mortality rates. Lipid metabolism abnormalities play a critical role in the pathogenesis of these diseases, yet their underlying molecular mechanisms remain incompletely understood. This innovative study systematically investigates the shared molecular mechanisms of lipid metabolism abnormalities in sepsis and atrial fibrillation using advanced bioinformatics and machine learning methodologies. Methods: We retrieved two independent research cohorts from the Gene Expression Omnibus database: sepsis-related datasets and atrial fibrillation-related datasets. A multi-dimensional analytical approach was employed, including differential expression analysis, Weighted Gene Co-expression Network Analysis, machine learning models, immune cell infiltration analysis, and gene set enrichment analysis to comprehensively elucidate the molecular mechanisms of lipid metabolism abnormalities in these diseases. Comprehensive analysis identified 13 key candidate genes, with CD81, CKAP4, and DPEP2 emerging as core characteristic genes. Functional enrichment analysis revealed these genes primarily participate in mitochondrial function regulation, complement-coagulation cascade, and cell adhesion molecular pathways. Machine learning models demonstrated exceptional diagnostic performance, with area under the curve values of 0.957 for sepsis and 1.000 for atrial fibrillation datasets. Immune cell infiltration analysis unveiled the critical roles of neutrophils and monocytes in disease progression, and revealed the profound impact of lipid metabolism abnormalities on immune regulation. We discovered that lipid metabolism abnormalities significantly modulate disease progression by influencing mitochondrial function, inflammatory responses, and cell adhesion pathways. Future research necessitates further clinical validation and functional experiments to explore personalized therapeutic strategies based on lipid reprogramming.