To investigate the mechanism of the ethyl acetate extract of Tumor Vine (PT-EAE) in combination with cisplatin (DDP) for the treatment of non-small cell lung cancer (NSCLC), using network pharmacology prediction and validation through cellular experiments. The chemical components of PT-EAE were identified using Ultra-Performance Liquid Chromatography-Quadrupole-Orbitrap Mass Spectrometry (UPLC-Q-Exactive-Orbitrap-MS). Network pharmacology was employed to predict the potential active components of PT-EAE and their molecular targets affecting NSCLC cell functions. A “PT-EAE-active components-targets-disease” interaction network was constructed. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on the potential targets to identify core targets and signaling pathways closely related to the active components of PT-EAE. Subsequently, in A549 cells, the effects of PT-EAE alone, DDP alone, and their combination (PT-EAE DDP) were evaluated. Cell Counting Kit-8 (CCK-8) assay, wound healing assay, transwell assay, and flow cytometry were used to detect cell proliferation, migration, invasion, and apoptosis, respectively. Finally, western blotting and quantitative reverse transcription polymerase chain reaction (RT-qPCR) were utilized to detect the expression levels and phosphorylation status of the predicted core molecular proteins following treatment with PT-EAE and DDP. PT-EAE inhibited the proliferation, migration, and invasion of A549 cells, while promoting apoptosis. Compared to the DDP monotherapy group, the PT-EAE DDP combination group reduced the IC50 of DDP for inhibiting A549 cell proliferation. Network pharmacology analysis identified 12 active components in PT-EAE, which were mapped to 124 NSCLC-related targets and multiple key signaling pathways. Among these, EGFR, Src, and PI3K were identified as potential core nodes. At the molecular level, PT-EAE downregulated the protein expression levels of EGFR and Src in A549 cells, as well as the phosphorylation levels of downstream molecules in the Src signaling pathway—PI3K, Akt, and mTOR. It also suppressed the mRNA expression of EGFR, Src, PI3K, Akt, and mTOR. The ethyl acetate extract of Tumor Vine, when combined with cisplatin, shows promising anti-tumor activity through multi-component, multi-target, and multi-pathway actions, with the PI3K/Akt/mTOR signaling pathway likely representing one of its therapeutic routes.
Milk proteins represent the principal protein constituents of mammalian milk and play indispensable biological roles in the growth, development, and immune maturation of neonates. Modulating the expression of genes involved in milk protein synthesis holds considerable promise for enhancing both the efficiency of milk protein production and the bioavailability of dairy-derived nutrients, thereby offering significant translational potential in neonatal nutrition and the functional optimization of bioactive milk peptides. This study conducts a comprehensive bibliometric analysis of the scientific literature on milk protein gene expression regulation, drawing upon publications indexed in the Web of Science Core Collection from 2006 to 2025. Utilizing CiteSpace and VOSviewer, we systematically map research hotspots, evolutionary trends, and emerging frontiers in the field. Concurrently, we provide an integrative review of the molecular mechanisms governing milk protein gene expression, including key transcriptional regulators and associated signaling pathways, as well as recent advances in the optimization and innovative applications of milk protein-based products and pharmaceuticals.
Clostridium perfringens and Clostridium botulinum are anaerobic spore-forming bacteria that produce potent toxins and are major causative agents of severe toxicoinfections in ruminants. Although vaccination remains the primary preventive strategy, current commercial vaccines are based on crude toxoid preparations, which present several limitations, including labor-intensive manufacturing processes, high biological risk, and batch-to-batch variability. This study aimed to evaluate the immunogenicity of a pentavalent recombinant bacterin vaccine formulation containing antigens derived from C. perfringens (CPA, CPB, and ETX) and C. botulinum (BoNT/C and BoNT/D) in cattle, sheep, goats, and buffaloes. Individual and chimeric recombinant antigens were expressed in Escherichia coli and incorporated into a vaccine formulation containing three inactivated bacterial components (bacterins) adsorbed onto aluminum hydroxide. A total of 92 ruminants (23 cattle, 23 buffaloes, 23 sheep, and 23 goats) were included in the study and allocated to receive the recombinant vaccine, a commercial vaccine, or saline as a negative control. Neutralizing antibody titers were determined by serum neutralization assays. The recombinant vaccine was well tolerated across all species and induced neutralizing antibody titers of 10 IU/mL against CPB, 5 IU/mL against BoNT/C, and 4 IU/mL against BoNT/D. The response to ETX was species-dependent, with cattle and sheep reaching antibody titers of 4 IU/mL, whereas buffaloes and goats reached 2 IU/mL. Neither the recombinant bacterin nor the commercial vaccine elicited detectable neutralizing antibody responses against CPA, highlighting the need for further strategies to improve the immunogenicity of this antigen. Nevertheless, the overall findings support the potential of the pentavalent recombinant vaccine to confer protective immunogenicity against multiple clostridial toxins. Incorporating recombinant nontoxic antigenic domains into a bacterin-based platform represents a safe, scalable, and antigen-sparing alternative to conventional toxoid vaccines, with promising implications for improving the prevention and control of clostridial diseases in ruminant livestock.
Bulbophyllum is a large and highly diverse genus of the family Orchidaceae. Investigations in the Northeast Indian region have revealed an abundance of various Bulbophyllum species. However, the taxonomy and phylogeny of these North-eastern species still remain unresolved. This study is the first comprehensive phylogenetic assessment of 20 species of Bulbophyllum from Northeast India, revealing new insights into their relationships. The complete nuclear ITS region (nrITS) (ITS1, 5.8S, ITS2) and the non-coding psbA-trnH intergenic spacer of plastid DNA, together with the morphometric characters were used to resolve the phylogeny of Bulbophyllum species of Northeast India. Individual as well as combined datasets were analysed to evaluate marker performance and overall resolution. The nrITS dataset showed better resolving ability as compared to plastid DNA marker. Phylogenetic trees generated from the combined analysis of morphological and molecular dataset provided the most robust phylogenetic framework, particularly at the sectional level. The results support the inclusion of B. reptans in the section Reptantia, rather than in Racemosae. The three species viz. B. odoratissimum, B. trichocephalum and B. cauliflorum were found to have a closer relationship suggesting that they belong to the same section. The section Desmosanthes needs further investigation to re-define its boundaries. Overall, this integrative approach improves the resolution of phylogenetic relationships within Bulbophyllum and provides a refined taxonomic framework. Furthermore, taxonomic key for the 20 species has also been developed to facilitate accurate identification, based on reliable diagnostic traits.
Osteoporosis (OP) is a metabolic condition primarily affecting older and postmenopausal women. It is characterized by decreased bone mineral density (BMD), reduced strength, and deteriorated bone microarchitecture. This drastically increases the risk of debilitating fragility fractures and chronic discomfort, severely harming quality of life. Current anti-OP medications can slow bone resorption but often fail to target bone tissue directly, resulting in significant systemic toxicity. A promising solution is nanodrug delivery designed to specifically target bones. This approach minimizes adverse effects by precisely transporting therapeutics to the skeletal site. Hydroxyapatite (HAP) is an ideal material for this purpose due to its close resemblance to human bone mineral. HAP-based nanosystems advanced treatment strategy by enhancing bone targeting, improving drug loading capacity, and optimizing release kinetics and biocompatibility. Furthermore, HAP composite scaffolds hold great promise for bone regeneration. Incorporating bioactive molecules and drugs into these composites facilitates the repair of bone defects. This local delivery method provides the necessary biological activity to directly enhance the bone regeneration process. This review explores recent advancements in HAP-based nanosystems for OP, focusing on their therapeutic mechanisms, interactions with bone tissue, clinical applications, underscoring their superior efficacy, and promising future. However, key challenges remain in translating these systems to clinical practice, including large-scale manufacturing, long-term safety evaluation, and the need for well-designed clinical studies to validate their therapeutic potential.
