Leukemia-secreted extracellular vesicles (EVs) carry biologically active cargo that promotes cancer-supportive mechanisms, including aberrant proliferative signaling, immune escape, and drug resistance. However, how antineoplastic drugs affect EV secretion and cargo sorting remains underexplored.

Elevated plasma homocysteine (Hcy) levels lead to hyperhomocysteinemia, a condition associated with various neurological disorders affecting multiple brain regions, including the hippocampus. In this study, we investigated the effects of exposing cultured rat hippocampal neurons to Hcy concentrations corresponding to mild, moderate, and severe hyperhomocysteinemia. A short 24-hour exposure had minimal effects, whereas prolonged exposure up to 14 days moderately enhanced hippocampal excitability without altering the gene expression of voltage-dependent calcium, sodium, or potassium channels or intracellular calcium levels. These findings suggest that Hcy-induced changes in neuronal excitability may contribute to neuropathologies associated with hyperhomocysteinemia.

Despite exhaustive research efforts, integrated genetic and epigenetic mechanisms contributing to tobacco-induced initiation and progression of lung cancers have yet to be fully elucidated. In particular, limited information is available regarding dysregulation of noncoding RNAs during pulmonary carcinogenesis.

The biofilm-mediated persistence of Pseudomonas aeruginosa in the food and biomedical sectors is currently a global concern. In light of this challenge, this study investigated a preventive approach against P. aeruginosa biofilm formation using Flavourzyme, a food-grade peptidase, considering its antibiofilm potential. The results revealed that a co-culture comprising 300 µL/mL (1 × minimum inhibitory concentration [MIC]) of Flavourzyme could kill P. aeruginosa. On the MBEC™ biofilm-forming device, 0.125 × MIC of Flavourzyme inhibited > 4.5 log CFU/peg of biofilm. Cell motilities and the biosynthesis of quorum sensing (QS) molecules such as N-acyl-homoserine lactones (AHLs), including C4-HSL, decreased significantly at 0.06 × MIC of Flavourzyme and became undetectable at 0.125 × MIC. Interestingly, while 0.03 × MIC of Flavourzyme elicited diverse expressions of QS and virulence-regulating genes, ≥ 0.06 × MIC of Flavourzyme remarkably suppressed the relative genomic expressions. This study proposes Flavourzyme as a potent antibiofilm agent against P. aeruginosa biofilms, recommending specific concentrations for effective use in food preservation.

Qiangguyin (QGY) is a traditional Chinese medicine prescription for postmenopausal osteoporosis. We modified the formula based on QGY and named it new QGY (N-QGY), and studied the therapeutic mechanism of N-QGY on osteoporosis.

The survival and adaptation of plants to adverse environmental conditions is crucial and is facilitated by receptor-like kinases, which act as cell surface receptors for a variety of signals. In this study, we identified a gene, GmHSL1b, encoding a receptor-like protein kinase that is responsive to abscisic acid (ABA) hormonal signals and is involved in the plant's response to drought and salt stresses. Subcellular localization assays have demonstrated that the GmHSL1b protein is located in the plasma membrane. Overexpression of the GmHSL1b gene in soybean enhanced root growth and development, as well as the plant's tolerance to salt stress, while the gmhsl1b mutant revealed increased sensitivity to salt stress. Comparative transcriptome analysis showed that some genes associated with various biological processes, such as mitogen-activated protein kinase (MAPK) cascade signaling, plant hormone signaling, cell wall remodeling, calcium signaling, and defense response mechanisms are differentially expressed in GmHSL1b overexpressing roots. Our research indicated that GmHSL1b can regulate the expression level of the candidate aquaporin GmPIP2-1, thereby affecting cell water content and the accumulation of reactive oxygen species (ROS) under salt stress. These findings indicate that the GmHSL1b participates in regulating root development and enhancing the tolerance to salt stress, thus offering insights for boosting crop adaptability to environmental stresses.

The mammalian circadian timing system is organized in a hierarchy, with the master clock residing in the suprachiasmatic nucleus (SCN) of the hypothalamus and subsidiary peripheral clocks in peripheral tissues. Because of the diversity of peripheral tissues and cell-types in the body, the existence of autonomous clock and identification of its potential entrainment signals need to be empirically defined on a cell type-by-cell type basis. In this study, we characterized the basic circadian clock properties of the adrenal zona glomerulosa cells, or ZG cells. Using isolated adrenal explants from Per2Luc mice, dissociated ZG cells from Per2-dluc rats, and a related human adrenocortical cell line H295R, we showed that ZG cells possess genetically-encoded, self-sustained and cell-autonomous circadian clock. As to the potential entrainment signals, angiotensin II (Ang II) caused phase-dependent phase-shifts of adrenal ZG cells in cultured slices. Ang II treatment also drove initiation (or reset) of circadian clock gene expression in H295R cells with associated immediate up-regulation of PER1 and E4BP4 mRNA expression. We found that the type I Ang II receptor blocker CV11974, one of the most widely used clinical drugs for hypertensive diseases, caused attenuation of the phase resetting of H295R cells. Our in vitro data provide a basis to understand and argue for the adrenal gland ZG cells as a component of autonomous and entrainable peripheral clocks.

Enterotoxigenic Escherichia coli (ETEC) is responsible for piglet diarrhea and causes substantial economic loss in the pig industry. Along with the restriction of antibiotics, natural compounds targeting bacterial virulence factors are supposed to be efficacious and attractive alternatives for controlling ETEC infection. This study aimed to investigate the influence of dihydromyricetin (DMY), a natural flavonoid compound, on the expression of virulence factors of ETEC and intestinal inflammatory injury.

Estrogen receptor (ER)-positive breast cancer is the most common subtype, accounting for approximately 80% of cases, with endocrine therapy as the standard postoperative treatment. However, despite risk-reducing therapies, the risk of recurrence remains substantial. Studies, including the POETIC trial, have demonstrated that low Ki67 levels following short-term neoadjuvant endocrine therapy (sNAET) are associated with a favorable prognosis. The objective of this study is to identify genes associated with the suppression of cell cycle progression by sNAET in postmenopausal patients with ER-positive/human epidermal growth factor receptor 2-negative breast cancer.

To explore the regulatory effect of c-Jun N-terminal kinase (JNK) inhibitor (SP600125) on forkhead box protein L2 (FOXL2) gene in human ovarian granulosa cell tumor cells (KGN cells). The main pathogenic gene FOXL2 of ovarian cancer was screened by bioinformatics method. KGN cells were randomly divided into control group and experimental group. Different concentrations of SP600125 (0.1, 1, 5, 10, 50 µM) were added to the experimental group, and an equal volume of dimethyl sulfoxide (DMSO) was added to the control group. The cells were incubated for 48 h. Cell RNA was extracted and reverse transcribed into cDNA. The mRNA expression level of FOXL2 was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Proteins were extracted, and the expression level of FOXL2 protein was detected by Western blot. The proliferation ability of KGN cells treated with SP600125 was detected by MTT assay. Cell scratch assay was used to detect its migration ability. Different concentrations of JNK inhibitor reduced the expression of FOXL2 in ovarian granulosa cells KGN, and 1 µM had the best inhibitory effect. JNK inhibitor reduces the expression of FOXL2 in ovarian granulosa cell tumor KGN.

Melanoma is among the 10 most prevalent malignant tumors, posing a significant threat to human health. A detailed understanding of the molecular mechanisms driving its progression is crucial for advancing treatment strategies and outcomes. Based on bioinformatic analysis and experimental validation, this study identified mitochondrial carrier homolog 2 (MTCH2) as a key regulator of melanoma proliferation. Mechanistically, MTCH2 enhanced the expression and nuclear translocation of nuclear factor (erythroid-derived-2)-like 2 (NRF2), which up-regulated ribonucleotide reductase subunit M1 (RRM1) expression, thereby promoting melanoma cell proliferation. Targeting RRM1 in combination with dacarbazine significantly inhibited tumor growth in nude mouse xenograft models. These findings elucidate a mechanistic link between MTCH2 and the NRF2-RRM1 axis in melanoma proliferation and highlight potential therapeutic targets for intervention.

Fluoxetine was reported to restore critical period-like neural plasticity via alleviating perineuronal nets (PNNs). This study aimed to investigate the effect of fluoxetine on auditory processing and PNNs in auditory cortex and hippocampus.

Pathogenic RNA splicing variants have emerged as promising therapeutic targets due to their role in disease while preserving coding sequences. In this study, we developed RECTAS-2.0, a small molecule designed to correct RNA mis-splicing caused by the GLA c.639+919G>A mutation, which leads to the inclusion of a 57-nucleotide poison exon, resulting in later-onset Fabry disease, particularly prevalent in East Asia. RECTAS-2.0 restored normal GLA mRNA splicing and α-galactosidase activity in patient-derived B-lymphoblastoid cell lines and induced pluripotent stem cell-derived cardiomyocytes. Furthermore, oral administration of RECTAS-2.0 effectively corrected splicing in a transgenic mouse model, demonstrating its substantial splice-switching activity and safety for clinical application. RECTAS-2.0 demonstrated potential applicability to other genetic disorders that involve similar exon competition. These findings underscore the therapeutic potential of RECTAS-2.0 for Fabry disease and highlight its broader implications for RNA splicing-targeted therapies in genetic disorders.

Gastric cancer is a highly heterogeneous and aggressive disease with limited treatment options, necessitating innovative therapeutic strategies. Drug repurposing, a cost-effective approach, offers opportunities to identify new applications for existing medications. This study systematically investigated the apoptotic effects of serotonergic drugs on MKN-45 gastric cancer cells, providing a novel perspective on serotonin signaling in cancer therapy.

Proteolysis Targeting Chimeras (PROTACs) represent promising therapeutic modalities for degrading disease-causing proteins. However, the development of effective PROTACs has been limited by the availability of suitable E3 ligase ligands. In this study, we demonstrate for the first time that SPOP, an unexplored E3 ligase, can be recruited to degrade target proteins of interest. We developed a bridged PROTAC strategy and successfully discovered a proof-of-concept PROTAC degrader 9 (MS479), which recruits the E3 ligase SPOP by directly binding its substrate GLP as a bridge protein. This approach facilitates the polyubiquitination and subsequent degradation of BRD4/3/2 by the 26S proteasome. 9 effectively reduced the protein level of BRD4 short isoform in a time-, concentration-, GLP-, SPOP-, and ubiquitin-proteasome system (UPS)-dependent manner. Additionally, 9 effectively inhibited the proliferation of colorectal cancer (CRC) cells. Overall, our study expands the limited repertoire of the E3 ligases that can be harnessed for targeted protein degradation.

Lidocaine is a commonly used local anesthetic that blocks sodium channels in nociceptor neurons, preventing the transmission of pain signals to the brain. Lidocaine can be administered to reduce discomfort during tissue biopsies. Biopsy tissue may then be used to study the transcriptome under the assumption that the genomic activity of lidocaine-treated tissue accurately reflects that of untreated tissue. This study investigated how intramuscular lidocaine injection influenced skeletal muscle gene expression in sheep, aiming to understand how transcriptomic changes could affect data interpretation. Approximately 10 min before euthanasia, the left biceps brachii muscle from each of 6 wether lambs (48.7 ± 0.8 kg) was injected (IM; 20G hypodermic needle) at a depth of 3 cm with 2 mL of 2% lidocaine (20 mg/mL); the right biceps brachii was untreated. At necropsy, muscle samples were collected from the injection sites and contralateral limbs and flash-frozen. In an additional set of lambs, lidocaine-treated and untreated samples were collected from the biceps brachii of 4 lambs, and the vastus intermedius of 4 other lambs. RNA was isolated and mRNA sequenced to a targeted depth of 20 million reads per sample. Sequences were mapped and quantified; matched-pair analysis was performed in EdgeR. No genes were consistently differentially expressed due to treatment in both muscle types, perhaps reflecting their distinct physiological roles. Lidocaine did influence the transcriptome with anti-inflammatory effects evident in both muscle types, including the downregulation of immune-associated transcription factors and other genes. Lidocaine's influence varied on other broad categories of genes, including those associated with muscle contractility, tissue repair, and structural integrity, which could affect the interpretation of transcriptome data in studies of muscle growth and development. Pathway analysis revealed that lidocaine impacted signaling mechanisms for cellular connectivity and structure. This study demonstrates that intramuscular administration of lidocaine results in the alteration of tissue's gene expression profiles, highlighting the importance of considering lidocaine's influence in transcriptome analyses. Thus, the use of complementary physiological measures to validate transcriptomic findings is recommended to ensure observed gene expression changes are accurately attributed to experimental conditions rather than the effects of lidocaine.

Vascular calcification contributes to morbidity and mortality in aging and is accelerated in diabetes and in chronic kidney disease. Matrix Gla Protein is a potent inhibitor of vascular calcification, which is activated by the vitamin K-dependent gamma-glutamyl carboxylase (GGCX). However, through a currently unidentified mechanism, the activity of GGCX is reduced in experimental uremia, thereby contributing to the promotion of vascular calcifications. In this study, we aim to identify the cause of these functional alterations and to stimulate the enzyme activity by potential GGCX binding compounds as a new avenue of therapy.

Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic kidney disease worldwide. The one small molecule drug available to patients, tolvaptan, is associated with off-target side effects and high discontinuation rates, necessitating the development of new therapeutic strategies. Previous work has shown that the epigenome is altered in ADPKD; however, the identification and targeting of dysregulated epigenetic modulators has yet to be explored for human ADPKD therapy. Using cells derived from cysts of ADPKD patients, we tested the gene expression of several epigenetic modulators. We found Brd4 and BMi1 are upregulated and observed that their inhibition using small molecule drugs, AZD-5153 and PTC-209, significantly slowed the proliferation of ADPKD patient cells. To enhance the delivery of AZD-5153 and PTC-209 to renal cells, we loaded the drugs into kidney-targeting micelles (KM) and assessed their therapeutic effects in vitro. Combining AZD-5153 and PTC-209 in KMs had a synergistic effect on reducing the proliferation in ADPKD patient cells and in a 3D PKD cyst model. These findings were also consistent in murine in vitro models using Pkd1 null renal proximal tubule cells. In summary, we demonstrate Brd4 and BMi1 as novel targets in ADPKD and targeting the epigenome using kidney nanomedicine as a novel therapeutic strategy in ADPKD.

The functional role of estrogen receptor activation factor (E-RAF), a 66 kDa protein in mediating the biological actions of estrogen and progesterone in goat uterus, has been highlighted in this work. Here we report that E-RAF stimulates the expression of splicing factor gene, arginine/serine rich in goat uterine nuclei (GenBank Accession No. MT473439). cDNA subtractive hybridization method using nuclear run-on transcription assay was used to identify the differentially expressed genes. The upregulation of the splicing factor gene was demonstrated further using a PCR-based nuclear run-on transcription system. Confocal microscopic analysis has confirmed the endoplasmic reticulum as the primary site of intracellular location of E-RAF. The effect of estrogen and progesterone on the intracellular distribution of E-RAF and the molecular mechanisms that underlie the transport of E-RAF to nucleus in the presence of progesterone has been studied. Our results offer a new perspective in steroid hormone action in uterus.

Despite the established efficacy of hydroxyurea (HU) in increasing fetal hemoglobin (Hb F) levels in patients with intermedia beta-thalassemia (β-thal) and sickle cell anemia, the precise molecular mechanisms underlying these effects remain largely elusive. Understanding these mechanisms is paramount for identifying alternative therapeutic approaches to increase Hb F production while minimizing adverse effects. In this study, we employed weighted gene co-expression network analysis (WGCNA) to investigate the molecular underpinnings of γ-globin switching within GSE90878 dataset. Leveraging this information, we aimed to predict the transcriptome network and elucidate the mechanism of action of HU and Metformin (Met) on this network comprehensively. Through bioinformatic analysis, we identified IGF2BP1 and GCNT2 as key regulators of the γ-globin switching mechanism. To experimentally validate these findings, we utilized the K562 cell line as an erythroid model. Cells were treated with HU (50, 100, and 150 µM) and Met (50, 100, and 150 µM) for 24, 48, and 72 h. The expression levels of the GCNT2, γ-globin, IGF2BP1, miR-199a/b-5p, miR-451-5p and miR-144-3p were quantified using real-time polymerase chain reaction (qPCR). Our results revealed that treatment with HU (150 µM), Met (100 µM), and combination of HU-Met (150/100 µM) significantly increased IGF2BP1 expression by 6.2, 5.3, and 7.1-fold, respectively, after 24 h treatment. Furthermore, treatment with HU (50 µM), Met (50 µM) and HU/Met (50/50 µM) for 24 h led to a 3.3, 1.2, and 5-fold decrease in GCNT2 gene expression, respectively. Notably, the highest levels of γ-globin expression and Hb F production were observed with HU (100 µM), Met (50 µM), and HU/Met (100/50 µM). This study provides compelling evidence that HU and Met significantly enhance γ-globin expression and Hb F production in the K562 cell line. Our findings suggest that these drugs exert their effects by modulating the expression of IGF2BP1 and GCNT2, thus offering valuable insights into potential therapeutic strategies for disorders characterized by low Hb F levels.