Sulfated polysaccharides from edible marine algae have gained significant attention for their remarkable nutritional and therapeutic potential. Caulerpa racemosa (sea grape), a widely consumed edible algae, is a source of bioactive compounds with significant medicinal potential. This study focuses on CRP-2, a sulfated (1 → 4) galactan derived from C. racemosa, and its anti-inflammatory effects in lipopolysaccharide (LPS)-induced CALU-1 cells. CRP-2, at a concentration of 125 μg/mL, reduced the expression of pro-inflammatory cytokines interleukin-1β and tumor necrosis factor-α by 41-49 % in CALU-1 cells. Additionally, it significantly downregulated transforming growth factor-β levels, as indicated by a decrease in mean fluorescence intensity from 20 to 4 units, compared to cells treated with LPS alone. Concurrently, downregulated expressions level of interleukin-33, interferon-α and interleukin-10 in LPS-induced CALU-1 cells were substantially elevated after the treatment with CRP-2 (mean fluorescence intensities 25-35-fold), in a dose-dependent manner. Gene expression analysis by qRT-PCR revealed downregulation of interferon-γ levels by 2 folds (at 125 μg/mL) from an overly expressed levels of 9-fold in LPS-induced cells. Structure-activity and molecular docking analyses emphasized the role of sulfate moieties in its bioactivity. These findings highlight the potential of CRP-2 as a natural anti-inflammatory agent and suggest its promise as a functional food ingredient with notable health benefits.

Astragalus injection, a solution derived from Astragalus mongholicus Bunge, has been used in cancer patients for its immune-boosting properties. The effects of Astragalus injection combined with immune checkpoint inhibitors (ICIs) in cancer remain unsuspected.

Swainsonine (SW), the primary toxic component of locoweed, induces toxic response in grazing livestock. The swainsonine induced toxicity is characterized by symptoms including head tremors, ataxia, and limb paralysis. The mechanisms of the toxicity remained to be investigated. The unfolded protein response (UPR) plays a key role in alleviating endoplasmic reticulum stress (ERS) by reducing protein synthesis and promoting the degradation of misfolded proteins. ERS is closely associated with both the UPR and autophagy activation. However, the involvement of the UPR signaling pathway in SW-induced ERS and autophagy remains unclear. In this study, we demonstrate that SW up-regulates the expression of GRP78, XBP1s, LC3-II/I, and ATG5 in both in vitro and in vivo models, suggesting activation of ERS, UPR, and autophagy. To investigate the molecular mechanisms by which the UPR regulates autophagy under ERS in primary rat renal tubular epithelial cells (RTECs), we observed that inhibiting PERK led to increased levels of p62. Inhibition of ATF6 significantly reduced the up-regulation of LC3-II/I, p62, and ATG5. Furthermore, inhibiting IRE1α significantly decreased the expression of LC3-II/I and p62. These findings suggest that PERK and ATF6 regulate autophagy mainly by modulating the expression of autophagy-related genes, while IRE1α likely regulates these genes through the IRE1α-XBP1 pathway. Additionally, autophagy is directly regulated through the IRE1α-JNK signaling pathway.

The aim of this study was to verify the hypothesis that kisspeptin, a peptide encoded by the kiss1 gene, regulates steroidogenesis and cell proliferation in ovarian granulosa cells (GCs) from Tan sheep through modulation of let-7b and ITGB7 (integrin subunit beta 7). First, primary ovarian GCs were transfected with let-7b mimics and inhibitors. Next, HEK293T cells were cultured to validate the targeting relationship between let-7b and ITGB7, followed by the overexpression and knockdown of ITGB7 in GCs. Finally, GCs were treated with the PI3K-AKT/ERK signaling pathway inhibitor and 500 nM kisspeptin after transfection with ITGB7. EdU assays, flow cytometry, quantitative PCR (qPCR) and Western blotting were then used to detect cell proliferation, cell cycle and apoptosis as well as related gene and protein expression. The results showed that let-7b significantly inhibited progesterone secretion and cell proliferation while promoting apoptosis in GCs by targeting ITGB7. Notably, overexpression of ITGB7 led to a marked upregulation of p-ERK/ERK, p-PI3K/PI3K, and p-Akt/Akt. Furthermore, co-treatment with kisspeptin and ITGB7 significantly enhanced progesterone secretion and cell proliferation while reducing apoptosis in ovarian GCs. These results provide novel insights into the mechanism by which kisspeptin downregulates let-7b and upregulates ITGB7, thereby promoting steroidogenesis and cell proliferation while inhibiting apoptosis via the ERK/PI3K-Akt signaling pathway in Tan sheep. This study provides new insights into the molecular mechanisms by which kisspeptin regulates the function of ovarian GCs, and may lay the foundation for the future development of new kisspeptin-mediated reproductive regulation techniques in Tan sheep.

Neuroblastoma (NB), the common extracranial solid tumor in children, is associated with a poor prognosis, particularly in high-risk patients. MYCN amplification stands as the most prominent molecular hallmark within this high-risk subgroup. However, MYCN protein is considered "undruggable" due to its lack of a conventional enzymatic binding pocket and its predominant nuclear localization, which precludes targeting by standard small-molecule inhibitors or antibody-based therapeutics. Consequently, current therapeutic strategies have achieved limited efficacy against MYCN-driven NB. Notably, MYCN not only orchestrates diverse metabolic reprogramming pathways in tumors but also exerts a pivotal influence on cellular differentiation. To overcome this therapeutic barrier, we seek to elucidate the contribution of purine metabolism to stemness maintenance in MYCN-amplified NBs and to discover novel small-molecule inhibitors capable of inducing differentiation in high-risk NBs.

Chemotherapeutic resistance is a major obstacle to chemotherapeutic failure. Cancer cell resistance involves several mechanisms, including epithelial-to-mesenchymal transition (EMT), signaling pathway bypass, drug efflux activation, and impairment of drug entry. P-glycoproteins (P-gp) are an efflux transporter that pumps chemotherapeutic drugs out of cancer cells, resulting in chemotherapeutic resistance. Several types of long noncoding RNA (lncRNAs) have been identified in resistant cancer cells, including ODRUL, MALAT1, and ANRIL. The high expression level of ODRUL is related to the induction of ATP-binding cassette (ABC) gene expression, resulting in the emergence of doxorubicin resistance in osteosarcoma. lncRNAs are observed to be regulators of drug transporters in cancer cells such as MALAT1 and ANRIL. Targeting P-gp expression using natural products is a new strategy to overcome cancer cell resistance and improve the sensitivity of resistant cells toward chemotherapies. This review validates the inhibitory effects of natural products on P-gp expression and activity using in silico molecular docking. In silico analysis showed that Delphinidin and Asparagoside-f are the most significant natural product inhibitors of p-glycoprotein-1. These inhibitors can reverse multi-drug resistance and induce the sensitivity of resistant cancer cells toward chemotherapy based on in silico molecular docking. It is important to validate that pre-elementary docking can be confirmed using in vitro and in vivo experimental data.

The p53 signaling pathway plays a critical role in regulating the cell cycle, apoptosis, and senescence, making it a key target in cancer research. The aim of this study was to investigate the effects of an ethanol extract from the stem of Ziziphus nummularia on the proliferation and expression of genes involved in the p53 pathway in MCF-7 breast cancer cells. To achieve this, real-time quantitative PCR (RT-qPCR) was used to evaluate the mRNA expression of downstream genes linked to cell cycle and senescence, including CycE or CCNEl, RBLl, and E2F1. Molecular docking simulations using Molegro Virtual Docker (MVD) were also performed to assess the potential inhibitory activity of metabolite compounds from Z. nummularia stem against p53-regulating kinase (TP53RK). The results showed that the IC50 value of Z. nummularia stem ethanol extract against MCF-7 cells was 38.27 ± 0.72 μg/mL. The results also revealed a reduction in the expression of downstream genes linked to cell senescence and the cell cycle: CycE or CCNE1 (p = 0.011), RBL1 (p = 0.008), and E2F1 (p = 0.005), which was observed through RT-qPCR analysis of mRNA expression. This fact indicated that the inhibitory effects on proliferation by the ethanol extract of Z. nummularia stem might occur via pathways associated with cell senescence and cell cycle arrest. Molecular docking results of metabolite compounds from Z. nummularia stem suggested that squalene (Rerank score -112.70 kJ/mol), and nummularine B (Rerank score -110.68 kJ/mol) had potential as TP53RK inhibitors. These Rerank scores were smaller compared to the Rerank score of adenyl-imidodiphosphate (AMP-PNP), which was the native ligand of TP53RK, as confirmed by molecular dynamics analysis. These in silico results were confirmed by the decrease in p21 (CDKN1A) mRNA expression. In conclusion, the anti-proliferative effects of the ethanol extract from Z. nummularia stem on breast cancer cells occurred by affecting cell cycle-related genes and inhibiting apoptosis protection mediated by overexpression of p21 (CDKN1A) through p53 activity.

Bacterial flagellin (flg22) induces rapid and permanent stomatal closure. However, its local and systemic as well as tissue- and cell-specific effects are less understood. Our results show that flg22 induced local and systemic stomatal closure in intact tomato plants, which was regulated by reactive oxygen- and nitrogen species, and also affected the photosynthetic activity of guard cells but not of mesophyll cells. Interestingly, rapid and extensive local expression of Ethylene response factor 1 was observed after exposure to flg22, whereas the relative transcript levels of Defensin increased only after six hours, especially in systemic leaves. Following local and systemic ethylene emission already after one and six hours, jasmonic acid levels increased in the local leaves after six hours of flg22 treatment. Using immunohistochemical methods, significant defensin accumulation was found in the epidermis and stomata of flg22-treated leaves and above them. Immunogold labelling revealed significant levels of defensins in the cell wall of the mesophyll parenchyma and guard cells. Furthermore, single cell qRT-PCR confirmed that guard cells are able to synthesise defensins. It can be concluded that guard cells are not only involved in the first line of plant defense by regulating stomatal pore size, but can also defend themselves and the plant by producing and accumulating antimicrobial defensins where phytopathogens can penetrate.

Plant viral diseases pose a significant threat to agricultural production, and the availability of effective drugs against viral diseases remains limited. In this study, we discovered that betulinic acid (BA), a pentacyclic triterpenoid derived from plants, delays infection by turnip mosaic virus (TuMV) in Nicotiana benthamiana. Transcriptomic analysis revealed that BA treatment specifically induced the expression of N. benthamiana phytosulfokine 3 (NbPSK3), a plant pentapeptide hormone with diverse functions, while TuMV infection suppressed its expression. Further study demonstrated that NbPSK3 positively regulates antiviral defence against TuMV infection. Disruption of PSK signalling by targeting the membrane-bound PSK receptors (PSKRs) promoted viral infection. Additionally, exogenous sulphonated PSK (active form) treatment significantly delayed infection by TuMV in N. benthamiana compared to unmodified PSK peptides (dPSK, inactive form) or control treatments, while silencing the receptor NbPSKR1 abolished the ability of PSK to inhibit TuMV infection. Moreover, the inhibition of TuMV infection by BA is dependent on the PSK-PSKR signalling pathway. Overall, these findings not only underscore the potential of BA as a promising and environmentally friendly agent for modulating plant viral diseases but also emphasise the role of the PSK signalling pathway in promoting at least partial resistance to TuMV, which might have interest for crop breeding.

Triple-negative breast cancer (TNBC) is a highly heterogeneous disease with limited treatment options and high relapse rates due to chemoresistance and the presence of cancer stem cells (CSCs). This study explores the molecular profile and invasive properties of two TNBC cell lines, MDA-MB-231 (Basal-Like 1; BL1 subtype) and HCC1806 (BL2 subtype), as well as their chemotherapy-resistant derivatives (doxorubicin and paclitaxel). Both cell lines exhibited CD44+ and CD24-/low profiles with significant differences in epithelial-mesenchymal transition (EMT) markers. Chemoresistant variants exhibited significant changes in CSC markers, EMT genes, and ALDH activity, particularly the upregulation of CD133, suggesting its role in chemoresistance. Analysis of embryonic pathways revealed a prominent role of Sonic Hedgehog signaling, particularly in the BL2 subtype. Resistant models also exhibited increased Notch receptor expression. This study also examined novel polyamine compounds with an amino-pyridine structure. These compounds showed significant cytotoxicity against both sensitive and resistant TNBC cells, enhancing the efficacy of standard chemotherapeutics (paclitaxel and doxorubicin). Additionally, they reduced stem-like properties and self-renewal capacity of CSCs. This comprehensive characterization of TNBC cell lines and their chemoresistant variants underscores the molecular heterogeneity of TNBC and highlights potential therapeutic targets and strategies to enhance treatment efficacy and overcome resistance.

Cadmium, a pervasive environmental pollutant, exerts detrimental effects on various tissues and cells, particularly targeting the reproductive system, thereby posing significant risks to both animal food safety and human health. Despite its widespread impact, research on substances capable of mitigating cadmium-induced reproductive toxicity remains scarce, especially concerning female reproductive health. Metformin, a widely used oral antihyperglycemic drug, has demonstrated a range of beneficial effects, including anti-aging and antioxidant properties. This study aims to investigate the potential and underlying mechanisms of metformin in alleviating cadmium-induced reproductive toxicity in females. Over a period of 35 consecutive days, mice were exposed to cadmium-contaminated water (32 mg/l) and orally administered 10 mg metformin dissolved in 0.2 ml normal saline. Our findings reveal that metformin effectively mitigates cadmium-induced disruptions in the estrous cycle, follicular development, and oocyte meiotic maturation. Specifically, metformin enhances ATP production in oocytes by boosting mitochondrial mass and biosynthesis, thereby counteracting cadmium-induced oxidative stress and spindle morphology defects during meiosis. Additionally, metformin restores the DNA repair capacity of oocytes, alleviating cadmium-induced DNA damage. This restorative effect is partially mediated by metformin's ability to improve key epigenetic modifications, such as histone acetylation, histone methylation, and DNA methylation in oocytes. These results underscore metformin's potential as a protective or therapeutic agent against cadmium reproductive toxicity, primarily by maintaining cellular homeostasis to bolster oocyte resilience against cadmium toxicity and preserving normal epigenetic modifications to ensure oocyte quality.

The functional characterization of promoter regions of CBDAS and PT genes of cannabinoids biosynthesis suggests that multiple factors including tissue-specific, phytohormones, and stress-related signals modulate their activity. Cannabis sativa L. has tremendous potential as a future crop for producing clinically important cannabinoid metabolites. While the cannabinoid biosynthetic pathway is largely known, the mechanistic details about its regulation are less understood. Decrypting the environmental and developmental factors regulating cannabinoid biosynthesis pathway may prove beneficial in pathway engineering and molecular breeding programs. Functional characterization of the promoter regions of key cannabinoid biosynthesis genes can provide useful insights into their transcriptional regulation. This study, therefore, is focused to uncover the role of different phytohormones and abiotic factors in influencing the activity of CsCBDAS and CsPT1 promoters through the development of promoter-GUS fusion expressing transgenic lines of Nicotiana tabacum. Spatial analysis across different tissues revealed that CsCBDAS and CsPT1 promoters drive a high level of GUS staining in leaf and flowers of the transgenic lines. A strong GUS staining was detected in the glandular trichomes of both tobacco transgenic lines. The results showed that out of the five hormones, three (IAA, GA3, and SA) and four (IAA, GA3, SA, and ABA) caused significant activation of CsCBDAS and CsPT1 promoters, respectively. While the light, heat, cold, salt, and wound stress induced promoter activity of both CsCBDAS and CsPT1, the drought stress was found to induce the activity of CsCBDAS promoter only. Validation of the expression patterns of these genes under different conditions in C. sativa through qRT-PCR suggested that phytohormones and abiotic factors may influence the cannabinoid biosynthesis in C. sativa by modulating their promoter activity.

Abscisic acid (ABA) regulates key plant processes, including seed germination, dormancy, and abiotic stress responses. While its physiological role in germination is well-documented, the molecular mechanisms are still poorly understood. To address this, we analyzed transcriptomic and metabolomic changes in ABA-treated germinating barley (Hordeum vulgare) embryos. To map ABA-responsive gene expression across embryonic tissues, we employed the Visium Spatial Transcriptomics (10× Genomics). This approach, which remains technically challenging to be applied in plant tissues, enabled the precise localization of gene expression across six embryo regions, offering insights into tissue-specific expression patterns that cannot be resolved by traditional RNA-seq.

In European seabass (Dicentrarchus labrax), dietary tryptophan (TRP) surplus has a notable modulatory effect on the hypothalamic-pituitary-interrenal axis under chronic stress and acute inflammation, affecting cortisol levels and neuroendocrine- and immune-related gene expression. A transcriptomic approach (RNA-seq) was applied to head-kidney samples of fish submitted to confinement stress and/or acute inflammation to uncover the biological mechanisms behind these effects. Undisturbed seabass fed dietary TRP supplementation showed an up-regulation of various innate immune functions, contrasting previous studies which indicated mainly a TRP regulatory role. Upon bacterial injection, TRP-fed fish showed a transcriptomic profile similar to their counterparts fed on control diet, indicating TRP's inability to modulate immune mechanisms under bacterial challenge. Under confinement stress, TRP-fed fish exhibited a molecular profile similar to unstressed control fish, highlighting TRP's role in mitigating stress. However, combining dietary TRP supplementation with confinement stress and immune stimulation by bacterial inoculation resulted in a unique molecular profile. Stressed fish fed TRP did not show the restorative effect of immune stimulation on carbohydrate metabolism and showed downregulated genes related to glycolysis and glycogenolysis. Additionally, transcription upregulation in these fish after bacterial injection included terms related to serine and steroid metabolism (carboxyl ester lipase 2), indicating tryptophan-induced changes in lipid mobilization in the head-kidney, potentially affecting cortisol synthesis and other hormones.

TNBC remains the most aggressive and therapy-resistant type of breast cancer, for which efficient targeted therapies have not been developed yet. Here, we identified TRPML3 (ML3) as a potential therapeutic target in TNBC. Our data showed that ML3 is significantly upregulated in TNBC cells compared with nontumorigenic control cells. ML3 knockdown (KD) impairs TNBC cell proliferation by inducing cell cycle arrest and caspase-dependent apoptosis. ML3 KD also inhibits TNBC cell migration and invasion. Mechanistically, ML3 KD reduces lysosomal number and enhances lysosomal acidification, which in turn activates mTORC1, thereby inhibiting autophagy initiation and flux. This disruption negatively impacts mitochondrial function, as evidenced by reduced ATP production, increased ROS and NO production, and mitochondrial fragmentation. Importantly, ML3 KD enhances TNBC cell sensitivity to doxorubicin and paclitaxel. The finding suggests that targeting ML3 disrupts lysosomal and mitochondrial homeostasis and enhance chemosensitivity, presenting ML3 as a potential therapeutic vulnerability in TNBC enhancing chemosensitivity.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide, with metastasis being one of the primary characteristics. Elemene, a natural product derived from Curcuma wenyujin, has been approved for treating advanced and metastasis HCC patients. However, the molecular mechanism of elemene in HCC is still unclear.

Bladder cancer (BLCA) is the most common malignant tumor in the urinary system, with a significantly higher incidence in men than in women, severely impacting quality of life. The STING1 gene (stimulator of interferon genes 1) plays a critical role in innate immunity by recognizing abnormal DNA and activating immune signaling pathways, promoting the expression of type I interferons and pro-inflammatory cytokines, thereby enhancing anti-tumor immune responses. Liquiritigenin (LQG), a flavonoid compound extracted from licorice, exhibits anti-inflammatory, antioxidant, and anti-cancer properties, capable of inhibiting tumor cell proliferation and invasion while regulating autophagy. This study aims to evaluate the role of LQG in regulating the STING1 gene and its anti-cancer mechanisms in bladder cancer.

Quercetin is a natural product with multiple activities, which possesses a promising antitumor effect on malignancies. The involvement of proline 4-hydroxylase II (P4HA2) in collagen synthesis is crucial in the growth of tumor cells. Apoptosis is a programmed cell death requisite for the stability of the intracellular environment. However, the relationship between quercetin and cell apoptosis, as well as the impact of P4HA2 in this connection, has not yet been specified in hepatocellular carcinoma(HCC).

This research aims to elucidate the mechanistic role of artesunate (ART) in enhancing the sensitivity of renal cell carcinoma (RCC) to sunitinib. To establish sunitinib-resistant RCC cell lines (786-O R and Caki-1 R), cells were treated with different concentrations of sunitinib and ART. The viability of the cells was measured through the cell counting kit-8 (CCK-8) assay. Tripartite motif-containing 24 (TRIM24) and paired box 6 (PAX6) expression were suppressed with lentiviral vectors, quantified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot (WB) analysis. TRIM24-PAX6 interaction was examined through co-immunoprecipitation (Co-IP) and deubiquitination assays. Additional assays included colony formation, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, WB detection of phosphorylated histone H2AX (γ-H2AX) for DNA damage, epithelial-mesenchymal transition (EMT) marker analysis, sphere formation, and stemness marker assessments. In vivo drug resistance was tested using a mouse subcutaneous xenograft model. ART enhanced sunitinib sensitivity in resistant RCC cells, reducing colony formation, inducing apoptosis, elevating γ-H2AX, and upregulating TRIM24. ART enhances sunitinib sensitivity in RCC cells by upregulating TRIM24 expression, which facilitates the ubiquitination of PAX6. This process leads to the suppression of EMT and stem cell-like transitions in RCC cells.

Bisphenol A (BPA) is a widely used industrial chemical with potential endocrine-disrupting effects on metabolic processes. This study investigates the impact of BPA on hepatic function and transcriptional regulation in mouse livers and AML12 cells. Male mice were exposed to low (5 g/kg) and high (50 g/kg) doses of BPA for six weeks. Transcriptomic analysis was performed on liver tissues, and histological examinations were conducted. AML12 cells were treated with varying BPA concentrations, and PPARG transcriptional activity was assessed using a luciferase reporter assay. Additionally, molecular docking, molecular dynamics (MD) simulations, drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), MM-PBSA calculations, and multi-species protein structure comparative analysis were employed to analyse the interaction between BPA and PPARG. Transcriptomic analysis revealed a decrease in differentially expressed genes with higher BPA doses, with low-dose exposure significantly downregulating hepatic Cpt1a mRNA levels. Histological examination indicated lipid vacuole formation at high doses without collagen deposition. BPA consistently inhibited PPARG activity in both MCF7 cells and mouse livers. BPA exposure disrupts hepatic lipid metabolism and PPARG activity, highlighting its role as an endocrine disruptor. Further research is needed to elucidate the long-term effects of BPA on liver health.