Prostate cancer (PCa) remains a significant global health challenge, necessitating innovative approaches for improved detection and treatment. Conventional methods often face limitations in specificity, sensitivity, and targeted drug delivery, leading to suboptimal outcomes and increased toxicity. Nanotechnology offers a promising avenue to overcome these hurdles, with nanoparticles (NPs) emerging as versatile tools for both diagnosing and treating PCa. This review explores recent advancements in nanoparticle technologies for PCa, focusing on their dual roles in early detection and targeted therapy. We highlight how the unique physicochemical properties of NPs enable enhanced drug loading, controlled release, and active targeting via specific ligands, thereby minimizing systemic toxicity and maximizing therapeutic efficacy. Furthermore, we discuss emerging applications of NPs as contrast agents for advanced imaging and as delivery vehicles for chemotherapeutics, gene therapies, and photothermal agents. Crucially, rather than merely describing preclinical progress, this review critically analyzes the significant bottlenecks in the translation of these nanoparticle-based theranostics from the laboratory to the clinical setting. By evaluating the challenges in scalable manufacturing, regulatory pathways, and long-term safety, we provide a comprehensive perspective on the steps required to realize the full clinical potential of nanomedicine in prostate cancer management.
The CRISPR-Cas9 system has revolutionized genome editing, but generating multiple gene knockouts (multi-KOs) remains challenging. This study presents an improved method for efficient generation of multi-KOs using sgRNA DNA strands and the CRISPR/Cas9 system. The method was tested on genes involved in the apoptosis pathway of HEK293T cells. The crRNA for each target gene was designed from Exon 1, near the starting codon ATG, such that a small deletion or insertion in this region would lead to a premature STOP codon and functional knockout. The T7 endonuclease I assay confirmed the high editing efficiency of sgRNAs in both single- and multiple-gene knockouts. Deep sequencing analysis revealed frameshift mutations and premature stop codons, indicating successful loss-of-function mutations. Using the sgRNA DNA strands provided a more uniform and efficient editing profile compared to other methods. This improved protocol offers a simpler, faster, and cost-effective approach for generating multigene knockout clones, with implications for functional genomics, therapeutic development, and synthetic biology.
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial inflammation and joint destruction. Oxidative stress has been implicated in its pathogenesis. This in vitro study aimed to investigate the effects of liensinine (LIE) on the apoptosis of fibroblast-like synoviocytes (FLSs) and its underlying mechanisms. RA-FLSs were isolated from patients and cultured in low-glucose Dulbecco's Modified Eagle Medium (DMEM). Following LIE treatment, cell viability was assessed using the cell-counting kit-8 assay, and lactate dehydrogenase (LDH) release was measured to evaluate cytotoxicity. Apoptosis was assessed by 4 ',6-diamidino-2-phenylindole (DAPI) staining and flow cytometry. Oxidative stress was evaluated by reactive oxygen species (ROS) production and mitochondrial membrane potential using 5,5 ',6,6 '-tetrachloro-1,1 ',3,3 '-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) staining. Caspase-3/7 activity was measured using the Caspase-Glo (R) 3/7 luminescent assay (Promega) following the manufacturer's instructions. RNA sequencing and network pharmacology analysis were performed to identify differentially expressed genes and signaling pathways affected by LIE. Western blotting was used to analyze key proteins in the mitogen-activated protein kinase (MAPK) and apoptosis pathways. Molecular docking and dynamics simulations evaluated LIE binding to P38 and Caspase9. LIE significantly reduced cell viability and increased LDH release in RA-FLSs in a dose-dependent manner. LIE induced apoptosis and elevated oxidative stress, evidenced by increased ROS and altered mitochondrial membrane potential. RNA sequencing and network pharmacology revealed significant changes in genes related to MAPK and apoptosis signaling. Western blot confirmed activation of p38 MAPK and caspase-3/9, suggesting LIE-induced apoptosis correlates with oxidative stress and MAPK pathway activation. LIE exhibited strong binding affinities to P38 and Caspase9, with favorable stability in dynamics simulations. These in vitro findings suggest that LIE has therapeutic potential for RA by targeting MAPK and apoptosis pathways in RA-FLSs, warranting further in vivo validation.
Mastitis caused by Staphylococcus aureus is a common inflammatory disease in mammals. However, the role of carnitine palmitoyltransferase 1A (CPT1A) in this process remains poorly understood. In this study, we examined the expression and function of CPT1A using both in vitro and in vivo models. HC11 cells were transfected with a CPT1A-overexpressing plasmid and then infected with S. aureus, with or without the NOD-like receptor family pyrin domain containing 3 (NLRP3) agonist BMS-986299. In vivo, we established a mouse mastitis model by injecting S. aureus into the mammary duct and achieved local CPT1A overexpression via AAV9 delivery. The results showed that S. aureus infection significantly downregulated CPT1A expression in mammary tissue and HC11 cells. CPT1A overexpression reversed the infection-induced decrease in cell viability, restored mitochondrial membrane potential, increased antioxidant enzyme activities, reduced malondialdehyde levels, and prevented the downregulation of Occludin and Claudin-3. Mechanistically, CPT1A overexpression was associated with enhanced NLRP3 succinylation and accelerated degradation, thereby inhibiting NLRP3 expression. Administration of the NLRP3 agonist BMS-986299 partially reversed the protective effects of CPT1A overexpression both in vitro and in vivo. Collectively, these findings demonstrate that CPT1A counteracts S. aureus-induced mastitis by promoting NLRP3 succinylation, which accelerates NLRP3 degradation, thereby alleviating mitochondrial oxidative stress and preserving blood-milk barrier integrity. Targeting the CPT1A-NLRP3 axis may represent a potential therapeutic strategy for S. aureus-associated mastitis.
Head and neck squamous cell carcinoma (HNSCC) presents a significant global health challenge, characterized by late-stage diagnosis and high rates of locoregional recurrence. Traditional tissue biopsy, while considered the gold standard, is invasive and often fails to capture the spatial and temporal heterogeneity of the tumor. Liquid biopsy has emerged as a transformative, noninvasive paradigm for molecular diagnostics, leveraging circulating biomarkers found in blood and saliva. This systematic review critically evaluates current evidence regarding the diagnostic accuracy, prognostic value, and molecular validity of circulating tumor cells, circulating tumor DNA (ctDNA), and the transcriptomic landscape of coding and noncoding RNAs (miRNAs, lncRNAs, and circRNAs) in HNSCC. Our review highlights the specificity of ctDNA for monitoring minimal residual disease, particularly in human papillomavirus- and Epstein-Barr virus-driven malignancies, where viral kinetics serve as sensitive biomarkers for recurrence. Furthermore, we explore the utility of "Saliva-Exosomics," suggesting that salivary exosomes may offer high sensitivity for early-stage oral cavity lesions due to anatomical proximity. The review also synthesizes data on novel RNA species, such as tumor-educated platelets and circular RNAs, which act as stable systemic biosensors. Despite promising results, significant heterogeneity in isolation protocols and detection platforms (e.g., next-generation sequencing vs. digital PCR) remains a barrier to widespread clinical implementation. We conclude that integrating multianalyte liquid biopsies into clinical practice holds the potential to personalize therapeutic strategies, improve survival outcomes, and redefine the standard of care in head and neck oncology.
