Current Medicinal Chemistry - Online First

Sepia Ink Polysaccharide (SIP) is a well-characterized, marine-derived glycosaminoglycan with demonstrated multifunctional properties; however, its pharmacological mechanisms remain unclear. This study aims to investigate the anti-tumor mechanism of SIP1 from Sepia esculenta ink in the treatment of triple-negative breast cancer (TNBC), with a focus on apoptosis and autophagy.

MDA-MB-231 cells exposed to cisplatin (CP) and SIP1 were assessed for apoptosis and autophagy by evaluating cell morphology, apoptosis and autophagy rates, and the expression of key genes involved in these processes using double staining, flow cytometry, and Western blotting.

The data revealed that SIP1 induced apoptosis in TNBC cells, as demonstrated by an increased apoptosis rate, an elevated expression level of the Caspase-3 protein, a decreased expression of Bcl-2, and an elevated Bax/Bcl-2 ratio. Additionally, SIP1 did not impact autophagy. CP induced both apoptosis and autophagy of breast cancer cells. The combination of SIP1 and CP exhibited synergistic effects, enhancing apoptosis by 2.33-fold compared to SIP1 alone and 1.25-fold compared to CP alone, while simultaneously reducing autophagy levels (0.84-fold compared to CP alone), as verified by the Beclin 1 protein content.

This work discovered that SIP1, a sulfated glycosaminoglycan with a low content of sulfate ester groups derived from Sepia esculenta ink, induced apoptosis by inhibiting autophagy, providing a novel perspective for a deeper understanding of the anti- tumor mechanism of SIP. Currently, the underlying molecular mechanisms by which SIP1 modulates the crosstalk between apoptosis and autophagy in TNBC cells remain unknown and require further investigation.

This study demonstrates that SIP1 is effective in inducing apoptosis and promotes cisplatin-induced apoptosis by repressing cisplatin-induced autophagy in MDA-MB-231 cells.

Exosomal microRNAs (miRNAs) have been identified as pivotal regulators in the progression of diverse oncogenic processes. However, the relationship between exosomal miRNAs and the clinicopathological characteristics of gastric cancer (GC) patients remains a subject of debate. The present study was designed to meticulously assess the link between exosomal miRNAs and GC through a meticulous meta-analysis and rigorous database validation.

The case-control studies about the relationship between exosomal miRNAs and GC were retrieved from CNKI, SinoMed, Embase, Web of Science, the Cochrane Library and PubMed database. The retrieval time was from inception to November, 2023. Two researchers independently screened the literature, extracted the data and evaluated the quality of the included studies. The meta-analysis of the included literature was conducted by the Stata 12.0 software. The database of Kaplan-Meier plotter predicted that the expression of miRNA was correlated with prognostic value in GC patients. The study protocol has been registered in PROSPERO (CRD42023490351).

A total of 24 studies, involving 3490 participants, were included in this analysis. The meta-analysis results indicated that there was no significant decrease in the incidence of clinicopathological parameters associated with exosomal miRNAs in GC patients. However, analysis of the Kaplan-Meier plotter database revealed that high expression levels of hsa-mir-134, hsa-mir-100, hsa-mir-552, hsa-mir-30a, and hsa-mir-23b were associated with poor prognosis in GC patients, with hazard ratios (HRs) of 1.45 (95% confidence interval [CI]: 1.06-1.99, p=0.021), 1.67 (95% CI: 1.23-2.27, p=0.00098), 1.63 (95% CI: 1.11-2.40, p=0.012), 1.56 (95% CI: 1.08-2.26, p=0.017), and 1.52 (95% CI: 1.12-2.06, p=0.0066), respectively.

These findings align with prior studies highlighting the role of specific miRNAs in tumor progression but diverge regarding their diagnostic utility for clinicopathological features. Future research should explore the functional mechanisms of these miRNAs in GC biology and validate their prognostic value in larger, diverse cohorts to inform personalized treatment strategies.

Circadian rhythm genes (CRGs) play a significant role in the pathogenesis of various cancers, yet their impact on bladder cancer (BC) remains to be fully elucidated. EZH2, as a potential oncological biomarker, lacks clear delineation regarding its prognostic significance in BC. Furthermore, the effect of anesthesia on circulating tumor cells (CTCs) in cancer patients is scarcely studied.

In this study, we developed a bioinformatics signature based on CRGs to assess the prognosis of BC patients and investigated the expression of EZH2 in BC and its correlation with patient outcomes through clinical sample analysis. Furthermore, we collected blood samples from BC patients before anesthesia and two hours post-anesthesia, enriched for CTCs, and analyzed the expression of EZH2 to evaluate the impact of anesthesia on the quantity of CTCs and their EZH2 expression status.

Our prognostic model identified EZH2 as a key determinant of BC prognosis, with the high expression of EZH2 significantly associated with poor patient outcomes. Experimental validation revealed a significant increase in the number of EZH2⁺ CTCs after anesthesia in BC patients. These findings suggest that anesthesia may facilitate BC metastasis by increasing the number of EZH2⁺ CTCs.

The findings highlight the prognostic value of CRGs and EZH2 in BC, providing new insights into tumor biology and metastasis. Furthermore, this study suggests anesthesia may influence tumor progression by modulating EZH2 expression in CTCs, underscoring the need for careful anesthetic selection in BC patients.

This study unveils the potential value of CRGs and EZH2 in the prognostic assessment of BC and reports for the first time that anesthesia may influence tumor metastasis by modulating the expression of EZH2 in CTCs. These results offer new biomarkers for the prognosis and treatment of BC and provide novel insights into the role of anesthesia in cancer metastasis.

Neurodevelopmental disorders (NDDs) represent a diverse and heterogeneous group of conditions, including global developmental delay (GDD), autism spectrum disorder (ASD), and neurodevelopmental encephalopathy with epilepsy (NDEE). While these disorders often share phenotypic similarities, their underlying genetic causes can vary widely, making clinical diagnosis challenging.

In this study, we performed whole-genome sequencing (WGS) on a family having an autosomal recessive neurodevelopmental disorder. The proband (II-2) underwent WGS, followed by variant filtering through an in-house bioinformatics pipeline. Sanger sequencing and 3D protein modeling were performed to confirm the pathogenicity of the identified variant.

A novel biallelic missense variant in the NAV3 (c.3430T>C; p.Ser1144Pro) was detected using WGS and Sanger sequencing. Subsequently, 3D protein modeling revealed significant alterations in the secondary structure of NAV3, indicating a potential pathogenic effect.

The identification of a novel biallelic missense variant in NAV3 adds a new layer to our understanding of its potential contribution to autosomal recessive neurodevelopmental disorders. This case expands the mutational landscape of NAV3 and underscores its emerging significance in neurodevelopment.

This study reports a novel NAV3 variant in association with autosomal recessive NDD, contributing to the growing body of evidence supporting the involvement of NAV3 in human neurodevelopment. Functional validation and identification of additional patients will be essential to establish definitive genotype-phenotype correlations and uncover the mechanistic pathways underlying NAV3-associated disorders.

Scar heterogeneity, encompassing normal scar (NS) and pathological scars [hypertrophic scar (HS) and keloids], emerges from the dynamic interplay between systemic immune responses and local tissue microenvironment, highlighting the urgent need for drugs targeting different types of scars through both dimensions.

Data from DECODE and EQTLGen databases were used as exposure variables at the protein and mRNA levels in the blood, and data from GTEx and ScQTLbase as exposure variables at the tissue and single-cell levels. Two sample Mendelian Randomization (MR) studies were conducted at the systemic, local, and single-cell levels. The outcome variables were based on the NS, HS, and keloid cohorts in the authoritative FinnGen database. The results were ascertained using seven MR methods, including inverse-variance weighting (IVW), Wald ratio, weighted median, weighted mode, simple median, MR-Egger, and Summary-data-based Mendelian Randomization (SMR). Single-cell RNA-seq data were leveraged to validate the expression profiles and functions of the drug targets.

NUDT2, ATXN3, OGN, UROS, and TSG101 were significantly associated with keloids, while PARK7 and MZT2A showed a significant correlation with HSs, and CDCP1 was significantly linked to NSs. Among them, RNA and protein expression levels of NUDT2 and PARK7 demonstrated significant positive associations with keloids and HSs, respectively, at the blood, skin, and single-cell levels. Functional analysis revealed that the higher expression of NUDT2 was associated with angiogenesis and the cellular response to hormone stimuli, whereas PARK7 was involved in the organization of collagen fibrils and the extracellular matrix structure. Moreover, single-cell sequencing confirmed the high expression of NUDT2 and PARK7 in keloids and HSs. These findings highlight their potential roles in both systemic and local scar pathogenesis and underscore their promise as therapeutic targets.

This study identifies scar subtype-specific targets, particularly NUDT2 and PARK7, expanding therapeutic candidates for scar management. Multi-ethnic cohort studies are warranted to validate target universality.

Collectively, we have identified eight drug targets, with NUDT2 and PARK7 in particular showing potential therapeutic value for keloids and HSs. Additionally, our results suggest the feasibility of both local and systemic drug administrations.

Atherosclerosis (AS) is prevalent among the elderly population and poses a significant global health burden. However, the precise underlying mechanisms linking aging and mitochondrial dysfunction in AS remain unclear.

Through comprehensive utilization of databases including the Gene Expression Omnibus (GEO), MitoCarta, Molecular Signatures Database (MSigDB), and Human Aging Genomic Resources (HAGR), we employed various bioinformatics methods to explore the possible function of EF-hand domain family member D1 (EFHD1). This included the functional enrichment analysis, immune cell infiltration, and the lncRNA-miRNA-EFHD1 network. The validity of EFHD1 was confirmed using additional datasets and through Receiver Operating Characteristic (ROC) curve evaluation. Lastly, in vitro experiments were conducted using THP-1 cells treated with oxidized low-density lipoprotein (ox-LDL) to validate the expression and function of EFHD1 through Western blot and real-time quantitative PCR analyses. Additionally, in vivo experiments were performed on ApoE^-/- mice exhibiting atherosclerotic phenotypes, utilizing immunofluorescence staining.

Totally seven genes associated with aging and mitochondrial function (ALDH3A2, UCP1, BCL2, EFHD1, AHCYL1, HTRA2, and ALDH9A1) were discovered in AS, with EFHD1 identified as the principal hub gene. Immune infiltration analysis indicated that EFHD1 was negatively associated with myeloid suppressor cells (MDSC), activated B cells, and natural killer cells. An evident decline in EFHD1 was noted in unstable or advanced plaques compared to stable or early plaques, accompanied by significant area under the ROC curve (AUC) values of 0.917 (GSE100927) and 0.933 (GSE41571). Moreover, we recorded a reduction in EFHD1 expression in AS tissues and macrophages treated with ox-LDL. Following the silencing of EFHD1, TNF-α and IL-1β decreased, while ALODA, PKM2, MMP-9, JAK2, and STAT3 levels were upregulated. Furthermore, levels of ATP and reactive oxygen species (ROS) were diminished, while calcium ions and mitochondria levels remained unchanged.

To date, the common pathogenic genes associated with aging and mitochondrial dysfunction in atherosclerotic disease have been scarcely investigated. Using bioinformatics approaches, we identified seven hub genes (ALDH3A2, UCP1, BCL2, EFHD1, AHCYL1, HTRA2, and ALDH9A1) related to mitochondrial function and aging. Among these, EFHD1 was determined as the final hub gene. As a calcium sensor, EFHD1 plays a pivotal role in regulating mitochondrial metabolism and has been implicated in the prognosis of various tumors. Our findings demonstrated that EFHD1 knockdown decreased the levels of pro-inflammatory cytokines, such as IL-1β and TNF-α, increased JAK2 and STAT3 protein levels, and elevated MMP-9 levels, all of which may contribute to the vulnerability and progression of atherosclerotic plaques.

Our research revealed a reduction in EFHD1 expression within atherosclerotic tissues, suggesting its potential role in inflammation and mitochondrial energy metabolism as a key regulator of the calcium signaling pathway. This discovery offers possible advancements in the early diagnosis and treatment strategies for AS.

The objective of this study is to obtain inhibitors against mTOR targets with virtual screening, dynamic simulation and bioactivity assessment. This pursuit aims to obtain a rapid and accurate method for the discovery of new mTOR inhibitors.

Firstly, the researchers obtained nearly 9000 compounds by using ROC-guided machine learning from a library of over 200000 compounds. Secondly, virtual screening was used to evaluate the affinity of 45 compounds. Further analysis was performed to identify 6 hit compounds. Simultaneously, MTT antitumor activity evaluation and kinase inhibition assays are conducted for the active compounds to discern the most promising candidates. Furthermore, AO staining and JC-1 assays are performed for the selected compounds. Simultaneously, MTT antitumor activity evaluation and kinase inhibition assays are conducted for the active compounds to discern the most promising candidates. Furthermore, AO staining, JC-1 and hemolytic toxicity evaluation assays are performed for the selected compounds.

The kinase assay demonstrates that these 6 compounds display greater sensitivity to mTOR than to PI3K. Among them, compounds AJ-292/12941271 and AG-205/12550019 show better activity against mTOR target than PI3K, with an IC50 of 2.55 and 4.48 μM, respectively. Additionally, the anti-proliferative activity of the six hit compounds was also considered. Compound AJ-292/12941271 shows the best anticancer activity against A549 cell lines with an IC50 value of 4.3 μM. Further analysis reveals that compound AJ-292/12941271 induces apoptosis in the A549 cell line in a concentration-dependent or time-dependent manner. Hemolytic toxicity evaluation suggests that the compound AJ-292/12941271 is safe for further in vivo study.

This research proposes that the fused method of ROC-based machine learning, virtual screening, and bioactivity evaluation could be used to discover novel mTOR inhibitors quickly and precisely.

Regulatory T cells (Tregs) play an important role in the tumor microenvironment (TME). Currently, there have been no studies of Treg-related genes (TRGs) in lung adenocarcinoma (LUAD).

We integrated the Cancer Genome Atlas (TCGA) dataset with the Gene Expression Omnibus (GEO) dataset and divided the TCGA-GEO dataset patient samples into different cohorts by unsupervised clustering analysis based on the expression of TRGs in LUAD. By analyzing the TME characteristics of different cohorts, we assessed immune cell infiltration and function. In addition, we constructed Cox risk proportional regression models based on TRGs to predict patient prognosis.

The results of unsupervised cluster analysis classified the TCGA-GEO dataset as “immune desert”, “immune evasion” and “immune inflammation”. Moreover, there was a significant survival differential among the three cohorts (p-value < 0.05). Based on the expression of 61 TRGs in LUAD, we screened TFRC, CTLA4, IL1R2, NPTN NPTN and METTL7A to construct a Cox risk proportional regression model to divide the TCGA-GEO dataset into a training cohort and a test cohort. Survival was significantly worse in the high-risk group than in the low-risk group in both the training and test cohorts (p-value < 0.05). Finally, the nomogram scoring system constructed by integrating the model risk scores with clinical parameters can well predict the 1, 3 and 5 year survival of patients.

In conclusion, based on our analysis of the TRGs of LUAD patients, we can classify the patient TME into different immune statuses, which provides insights into adopting appropriate treatment regimens for different patients.

In 2022, the World Health Organisation (WHO) announced new cases of the developing Monkeypox Virus (MPXV), a zoonotic orthopoxvirus viral infection that mimics smallpox signs. Despite the ongoing infection, no proper medication is available to completely overcome this infection.

The study aims to construct a multi-epitope vaccine targeting Monkeypox Virus (MPXV) membrane glycoprotein to provoke robust immune responses.

To construct a potential immuno-dominant epitope vaccine to combat MPXV.

The target sequence, sourced from the UAE-to-India travel case, was analyzed to identify potential B-cell and T-cell epitopes (MHC-I and MHC-II). Immunodominant epitopes were selected and fused with β-defensin-I and PADRE to increase immunogenicity. The vaccine was modeled, docked with TLR3, and subjected to a 500 ns molecular dynamics simulation for stability analysis. Immune responses and bacterial expression were also evaluated.

The vaccine, comprising 230 amino acids, demonstrated antigenicity (0.6620), non-allergenicity, and broad population coverage. Selected epitopes included 3 B-cells, 4 MHC-I, and 2 MHC-II, ensuring a potent immunodominant profile. Docking with TLR3 revealed a binding affinity of -17.2 kcal/mol, while simulations confirmed their stability. Cloning (pET28a (+)) and immune response analyses showed a strong immunogenic profile, including elevated IgG1, IgM, and antigen levels, supported by a Codon Adaptation Index (CAI) of 1.0.

The proposed multi-epitope vaccine shows promise against MPXV. However, further in vivo and in vitro investigations are essential to confirm its immune efficacy.

Endoplasmic reticulum stress (ER stress) plays a crucial role in influencing the malignant behaviors of various tumors. Targeting the expression or degradation of transcription factors (TFs) offers a promising avenue for cancer treatment. However, a detailed understanding of how ER stress affects TF function and their interactions remains limited. This study aims to develop a prognostic model and identify TFs associated with ER stress in pancreatic ductal adenocarcinoma (PDAC).

We obtained gene expression profiles and corresponding clinical data from The Cancer Genome Atlas (TCGA). To develop a prognostic signature, we performed several analyses, including unsupervised clustering, enrichment analysis, immune infiltration assessment, as well as univariate, LASSO, and multivariate Cox regression analyses. Four transcription factors—STAT1, IRF6, NRF1, and RXRA—were incorporated into a risk model, which was subsequently validated using the GSE dataset. Additionally, we examined IRF6 through quantitative PCR, western blotting, flow cytometry, and immunohistochemistry in vitro using pancreatic cancer cell lines and a tissue microarray.

The high-risk group identified by the model exhibited significant associations with immune cell infiltration and poorer survival outcomes, though there was no significant correlation with tumor purity (p = 0.19). Furthermore, IRF6 downregulation in vitro was found to inhibit pancreatic cancer cell proliferation and promote apoptosis. IRF6 depletion also increased the expression of key molecules involved in ER stress at both the transcriptional and translational levels. Immunohistochemical analysis revealed marked differences in IRF6 expression between tumor and adjacent non-tumor tissues (59.29±29.88 vs. 95.22±40.80, p<0.001).

This study provides evidence that the constructed risk model can effectively predict prognosis in PDAC patients. Transcription factors related to ER stress, such as IRF6, show promise as both prognostic biomarkers and potential therapeutic targets for PDAC.

Ciprofol is a novel sedative-anesthetic that functions similarly to propofol. This study aimed to evaluate the efficacy and safety of ciprofol for the induction and maintenance of general anesthesia in patients undergoing thoracoscopic surgery.

A total of 120 patients undergoing thoracoscopic surgery for pulmonary nodules under general anesthesia were randomly assigned to the ciprofol group or the propofol group. Patients in the ciprofol group received an initial dose of 0.4 mg.kg^-1 of ciprofol for anesthesia induction, followed by an infusion rate ranging from 0.4 mg.kg^-1.h^-1 to 2.4 mg.kg^-1.h^-1 for maintenance. In the propofol group, patients were administered an initial dose of 2.0 mg.kg^-1 of propofol for induction, with a maintenance infusion rate ranging from 4.0 mg.kg^-1.h^-1 to 12 mg.kg^-1h^-1. The primary outcome measured was the success rate of sedation. Secondary outcomes included the time to successful induction of anesthesia, changes in hemodynamics and bispectral index (BIS) within 10 minutes after the initial administration of the study medication, time to respiratory recovery and full alertness, and the incidence of adverse events.

The sedation success rate was 100% in both groups. In this study, statistical analyses revealed no significant differences in the time to eyelash reflex disappearance (p=0.599), induction success time (p=0.431), the moment when the BIS value first fell below 60 (p=0.538), the time to respiratory recovery (p=0.505), or the interval until full wakefulness (p=0.837). Notably, within the first 10 minutes following the initial administration of the study medication, the reduction in blood pressure was significantly more pronounced in the propofol group (p<0.05). Additionally, the mean BIS value was significantly higher in the propofol group (p<0.01). The required dosage of sedative medication was significantly lower in the ciprofol group (p<0.001). Compared to the propofol group, the ciprofol group exhibited a significant reduction in the incidence of adverse events during intubation (p=0.01), a marked decrease in injection pain (p=0.001), and a significant decrease in the incidence of intraoperative hypotension (p<0.05).

Ciprofol exhibits comparable efficacy and safety profiles for both the induction and maintenance of general anesthesia in patients undergoing thoracoscopic surgery. Furthermore, it has been associated with a reduced dosage requirement, significantly lower mean BIS values, and a notable decrease in the incidence of injection pain and intraoperative hypotension.

(ChiCTR2400086976).

The pathogenesis of Delayed Encephalopathy After Acute Carbon Monoxide Poisoning (DEACMP) remains mysterious, and specific predictive markers are lacking. This study aimed to elucidate the molecular underpinnings and identify predictive biomarkers of DEACMP through multi-omics and single-nucleusRNA sequencing (snRNA-seq).

Clinical data and blood samples were collected from 105 participants. Untargeted metabolomics sequencing was employed to profile serum metabolites across these participants. Additionally, individuals from the Healthy Controls (HCs), Acute Carbon Monoxide Poisoning patients (ACOP), Non-Delayed Encephalopathy After ACOP (DEACMP-N), and DEACMP groups (n=3 each) were randomly selected for transcriptome sequencing to identify potential predictive targets and pivotal signaling pathways associated with DEACMP. Furthermore, Severe DEACMP and Control rat models were established. Three rats from the Control, DEACMP, and DEACMP + Dexamethasone + Selenomethionine groups were selected for snRNA-seq. Immunofluorescence multiplexing and qRT-PCR (quantitative Reverse Transcription Polymerase Chain Reaction) were then performed to validate the identified predictive targets.

Analysis of clinical data from 105 participants highlights the pivotal role of inflammation in influencing the prognosis of carbon monoxide poisoning. Metabolomics analysis identified 19 metabolites that significantly differed between the DEACMP-N and DEACMP groups. Transcriptomics analysis of 12 participants indicated that DEACMP is primarily associated with six signaling pathways, including lysosome and tuberculosis. Considering that microglia are central nervous system immune effectors, the snRNA-seq analysis revealed altered gene expression and signaling pathways in microglia during DEACMP, with KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis highlighting neutrophil extracellular trap formation, lysosome, and tuberculosis as the predominant pathways. Differential gene analysis from transcriptome and snRNA-seq identified 28 genes differentially expressed in DEACMP. The STRING (Search Tool for the Retrieval of Interacting Genes/Proteins) database, immune multiplexing, and qRT-PCR confirmed the pivotal role of the Ifngr1/Stat1/Ctss axis in DEACMP.

This research identifies the Ifngr1/Stat1/Ctss axis as a key inflammatory mechanism in the pathogenesis of DEACMP, thereby clarifying previous uncertainties regarding the sequelae of carbon monoxide poisoning. The intersection of lysosomal and tuberculosis pathways, as revealed through metabolomic, transcriptomic, and single-nucleus RNA sequencing analyses—especially within microglia—offers novel mechanistic insights that could inform therapeutic interventions. While the integration of multiple omics methodologies enhances the robustness of these findings, their biological relevance to the pathogenesis of DEACMP requires rigorous validation through independent cohort verification approaches.

This study provides a comprehensive overview of serum metabolite expression, differential gene expression, and signaling pathways in DEACMP, offering a theoretical foundation for understanding the pathogenesis of DEACMP.

Human oncostatin M (OSM) is a pleiotropic cytokine that regulates inflammatory and immune responses by binding to the heterodimer receptor complex OSM receptor beta (OSMRβ) and glycoprotein 130 (gp130). The distinct signaling pathways triggered by OSM are involved in multiple chronic inflammatory conditions, such as inflammatory bowel disease (IBD), rheumatoid arthritis (RA), and cancers, making the OSM-bound receptor complex a significant therapeutic target. Currently, no 3D structure of human OSM recognition complex is available, and thus, the molecular mechanisms underlying OSM signaling remain poorly understood.

In this study, for the first time, we proposed a full-length structural model of the human OSM-OSMRβ-gp130, generated using AlphaFold2 protein structure prediction and all-atom molecular dynamics (MD) simulation (~ 1.12 million atoms with explicit solvent), enabling investigation of the geometric and dynamic profiles of OSM-OSMRβ-gp130 structure at atomic-level.

Analysis of the simulation trajectory demonstrated that the structural rearrangements of the heterodimer receptors (i.e., OSMRβ and gp130) initiated by OSM binding mediated the signal transduction from the extracellular to the intracellular domains. In the representative conformation identified through clustering analysis, two main allosteric pathways contributed were found to mediate signal transduction from the allosteric region of OSM to the active sites of OSMRβ and gp130. Finally, two druggable binding sites located on OSM and gp130 were detected by dynamically monitoring pocket flexibility throughout the simulation. A comprehensive analysis of the OSM-OSMRβ-gp130 model was carried out with respect to OSM signaling.

The findings of this study not only enhance the mechanistic understanding of OSM binding to the heteromeric OSMRβ/gp130 but also identify druggable binding sites for structure-based design of small molecules to inhibit the intracellular signal transduction.

Staphylococcus aureus infections have become a significant public health issue due to increasing the resistance against known antibiotics, especially by Methicillin-Resistant Staphylococcus aureus (MRSA). Fluoroquinolones are broad-spectrum class of antibiotics mostly utilized in treating various bacterial infections and those caused by S. aureus. Reported data indicated that mutations of Ser84 to Leu, Ser85 to Pro and Glu88 to Lys in DNA gyrase A enzyme are the major cause of fluoroquinolone resistance against S. aureus. Therefore, the development of a novel targeted molecule with potential activity against mutant S. aureus is essential. The antibacterial activity of quinoline-clubbed hydrazone derivatives against S. aureus is noteworthy. However, the mechanism of action of quinoline hydrazone derivatives has not been reported by inhibiting these common mutations of DNA gyrase A.

In this concern, some quinoline hydrazone derivatives as antibacterial agents reported by several research groups have been further studied as mutated S. aureus DNA gyrase A (Pdb id: 8bp2) inhibitors using in-silico techniques viz., molecular docking, MD simulation, DFT analysis, and ADMET predictions.

Among the studied compounds, 42, 43, 48 and 49 were found to be the most active and showed the highest docking score (-7.71 to -9.29 kcalmol^-1) by interaction with mutant (Leu84 and Pro85) S. aureus DNA gyrase A. Further, MD simulation results indicated that these compounds exhibited good stability with the targeted macromolecule under dynamic conditions. The most active compound 49 (ʌE = 0.159 eV) attributed to its lower HOMO-LUMO gap, which was an indicator of a potential inhibitor of fluoroquinolone- resistant S. aureus DNA gyrase A enzyme. ADMET prediction study emphasized that both compounds showed a significant safety profile.

The future perspective emphasized that compounds 42, 43, 48 and 49 could be developed as novel inhibitors against fluoroquinolone-resistant DNA gyrase A enzyme on the completion of drug discovery approaches.

Nasopharyngeal carcinoma (NPC) is an aggressive malignancy with a poor prognosis. Ubiquitination is a complex post translational modification involved in cancer progression. However, ubiquitination related genes (URGs) in immunotherapy of NPC remains largely unexplored.

Differentially expressed URGs were screened based on the single-cell RNA sequencing (scRNA-seq) dataset and a risk model of NPC was constructed and evaluated for prognostic significance. The oncogenic role of RNF149 in NPC was investigated through in vitro and in vivo experiments, including tumor cells, NPC-like organoids, and tumor-bearing mice.

scRNA-seq data showed that URGs scores were higher in cancer cells than in normal epithelial cells. We identified 216 differentially expressed URGs between cancer and normal epithelial cells, but only 33 differentially expressed URGs associated with prognosis. Based on 33 URGs, TCGA-HNSC samples were classified into two distinct subtypes with significant differences in the tumor immune microenvironment, immunotherapy effect, and survival-prognostic genes. Using LASSO algorithm, 13 URGs were selected to construct a risk model, which demonstrated high predictive performance. The expression profiles of these 13 URGs were analyzed in TCGA-HNSC tumor and adjacent non-cancerous samples, and six URGs (BSPRY, OTUB1, PJA1, RNF149, RNF181, USP10) exhibited consistent expression trends. Moreover, quantitative real- time PCR revealed that RNF149 was up-regulated expression in NPC cells compared to the NP69 cells. RNF149 knockdown significantly impeded the proliferative, migratory, and invasive capabilities and exaggerated apoptosis of NPC cells. RNF149 knockdown cells exhibited a reduced capacity to form NPC organoids in a 3D culture system. shRNA-RNF149 diminished subcutaneous tumorigenic capacity of HK-1 cells compared to the control group.

The URGs-based prognostic risk model offers a robust tool for predicting immunotherapy efficacy in NPC and RNF149 promotes NPC progression.

A URGs-related prognostic risk model capable of predicting clinical outcomes in NPC patients and RNF149 promotes NPC progression. Our findings are expected to provide new strategies to improve outcomes for NPC patients.

Ultra-rare diseases (URDs) are defined based on point prevalence and are classified as conditions affecting fewer than 1 in 50,000 individuals, and they are more likely to exist among communities with higher consanguinity rates requiring evidence-based data.

In this multi-center study, we used next-generation sequencing to identify 30 pediatric patients with URDs. Along with the demographic information about their parents, clinical, laboratory, and radiological data was also obtained. Multinomial regression was carried out to assess statistical differences and determine associations using the Quality of Life of Childhood Epilepsy (QOLCE)-55 scale.

There were 19 male (63.33%) and 11 (36.67%) female patients. Their current age range was 2-15 years (mean=8.83 years). The majority were diagnosed with sodium channelopathy (64.51%). The average Quality of Life (QoL) score of all participants was 51.43 ± 9.01 (reference range 0-100) with quartiles Q1=40, Q2=43.5, and Q3=56.

We propose that URDs complicated by epilepsy can significantly impair the QoL of patients and their families.

The cyclin-dependent kinases (CDKs) play a crucial role in the normal progression of these stages. In tumor cells, CDKs are often highly expressed, leading to uncontrolled cell proliferation. Inhibiting the activity of CDKs in tumor cells can inhibit their growth and proliferation, thereby achieving anti-tumor effects. In recent years, many CDKs inhibitors have been developed, but due to side effects and drug resistance issues, only a few CDKs inhibitors have been approved by the FDA.

Publications on CDK-based dual-target inhibitors were reviewed using SciFinder and PubMed, excluding reviews, patents, and studies with irrelevant content.

The study outlines advancements in CDK-based dual-target inhibitors as antitumor agents, offering insights to support the development and application of more effective cancer therapies.

Dual-targeted anti-tumor drugs may have better therapeutic effects than single-targeted drugs, which may address drug resistance issues and overcome drug interactions and pharmacokinetic issues associated with combination therapy. As an important direction in cancer treatment, dual target inhibitors have broad development prospects. By continuing to explore and improve dual target therapies, it has potential to overcome many limitations of single target therapy and provide more effective and lasting treatment outcomes for cancer patients.

Glioblastoma is the most common and aggressive brain tumor, with low survival rates and high recurrence rates. Therefore, it is crucial to understand the precise molecular mechanisms involved in the oncogenesis of glioblastoma.

To investigate the regulatory mechanisms of long non-coding RNA (lncRNA)-microRNA (miRNA)-messenger RNA (miRNA) network related to glioblastoma, in the present study, a comprehensive analysis of the genomic landscape between glioblastoma and normal brain tissues from the Gene Expression Omnibus (GEO) dataset was first conducted to identify differentially expressed genes (DEGs) in glioblastoma. Following a series of analyses, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, protein-protein interaction (PPI), and key model analyses. In addition, we used the L1000CDS2 database bioinformatic tool to identify candidates for therapy based on glioblastoma specific genetic profile.

In our results, 100 key genes, 50 upregulated and 50 downregulated, were ultimately identified. The results of KEGG pathway enrichment gene analysis showed that the five regulatory pathways. Furthermore, 3 small molecule signatures (trichostatin A, TG-101348, and vorinostat) were recommended as the top-ranked candidate therapeutic agents. Nevertheless, the constructed miRNA-mRNA network revealed a convergence on 40 miRNAs. We found that dysregulation of lncRNAs such as KCNQ1OT1 and RP11-13N13.5 could sequester several miRNAs such as hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-106a-5p, etc., and promote the development and progression of glioblastoma.

Our study identified key genes and related lncRNA-miRNA-mRNA network that contribute to the oncogenesis of glioblastoma.

Hepatocellular carcinoma (HCC) has a poor prognosis due to late diagnosis and rapid progression, highlighting the need for a deeper understanding of its pathogenesis. Lysine crotonylation (Kcr), a unique post-translational modification, plays a crucial role in epigenetic regulation. However, the role of crotonylation-related genes (CRGs) in HCC remains poorly understood, necessitating an investigation of their prognostic and therapeutic relevance.

Transcriptomic and clinical data were obtained from TCGA and GEO databases. A CRG-based risk score was developed using Cox and LASSO regression analyses. To enhance survival prediction, a nomogram incorporating the risk score was constructed. Immune cell infiltration and drug sensitivity were assessed using CIBERSORT and 'OncoPredict.' Single-cell sequencing was employed to examine CRG expression within the HCC tumor microenvironment.

An 8-gene risk score model (HDAC2, ACADS, HDAC1, ENO1, PPARG, ACADL, ACSL6, and AGPAT5) was established, effectively stratifying patients into high- and low-risk groups in the training set. Cox regression and Kaplan-Meier analyses validated its prognostic value in the test set. The nomogram demonstrated enhanced prognostic accuracy for survival prediction. Differences in immune cell infiltration and immune checkpoint expression between risk groups highlighted the association between CRGs and the tumor immune microenvironment. Single-cell sequencing revealed that CRGs were highly expressed in key immune cells within the HCC microenvironment. Additionally, drug sensitivity analysis suggested that specific targeted therapies may be more effective in HCC patients.

Crotonylation-related gene signature demonstrates strong prognostic value in hepatocellular carcinoma (HCC), effectively stratifying patients into high- and low-risk groups and recapitulating known oncogenic roles of HDAC1/2, ENO1, PPARG, AGPAT5 and the protective functions of ACADS, ACADL, and ACSL6. It was found that crotonylation not only influences tumor cell metabolism and epigenetic regulation but also shapes the immune microenvironment, highlighted by distinct checkpoint expression, differential immune cell infiltration, and drug sensitivity profiles, which position our model as a promising tool for personalized therapeutic decision-making. However, clinical translation will require standardized, reproducible assays for crotonylation measurement and rigorous validation across diverse HCC etiologies (e.g., viral vs. non-viral), along with mechanistic and longitudinal studies to dissect causality versus correlation, assess off- target effects of crotonylation modulators, and confirm functional impacts on immune modulation before routine diagnostic or therapeutic use.

This study identifies a prognostic CRG signature for HCC and provides novel insights into personalized treatment strategies.

The incidence of psychiatric disorders, such as post-traumatic stress disorder (PTSD), anxiety, and depression, has been steadily increasing, while current treatment approaches remain limited in efficacy. As a result, there is an urgent need to explore more effective therapeutic interventions. In recent years, MDMA (3,4-methylenedioxymethamphetamine)-assisted therapy (MDMA-AT) has emerged as a promising and innovative approach, demonstrating favorable clinical potential in the treatment of these disorders. Although preliminary studies have confirmed its therapeutic efficacy, a comprehensive and systematic analysis of the research trends and current limitations of MDMA-AT remains lacking.

This study employed a bibliometric approach to systematically retrieve and analyze research literature published between 1994 and 2023 on the application of MDMA in the treatment of PTSD, anxiety, and depression. Relevant data were obtained from three prominent databases: Web of Science Core Collection, PubMed, and Scopus. VOSviewer and Microsoft Excel were used to perform visual and quantitative analyses, focusing on publication trends, research hotspots, prolific authors, leading institutions, and international collaboration networks.

The findings indicated a substantial increase in MDMA-related research over the past decade. The United States has led the field in publication output, with the Multidisciplinary Association for Psychedelic Studies (MAPS) identified as the most productive institution. Key figures, such as Rick Doblin, have demonstrated high influence and centrality within the global research network. The research focus has gradually shifted from investigations of the neurotoxic properties of MDMA to explorations of its therapeutic mechanisms, safety profiles, and clinical applications.

This study provides a comprehensive synthesis of the past thirty years of research on MDMA-AT in the treatment of PTSD, anxiety, and depression, identifying major research trajectories and critical challenges in the field. While current findings highlight the therapeutic promise of MDMA and its translational potential, further research is needed to improve trial design, enhance sample diversity, and evaluate long-term effects in order to support the standardization and evidence-based integration of MDMA-assisted therapy into clinical practice.

Fructose, like other sugars and sugar metabolites, is capable of glycating protein. Insulin's fructosylation leads to the generation of Advanced Glycation End Products (AGEs). Reducing sugars reaction with proteins to form Schiff’s bases, which are characterized by the presence of an imine (C=N) bond. The Schiff bases then undergo irreversible rearrangements, followed by the production of much more stable compounds called Amadori products. These Amadori products can further undergo oxidation, dehydration, cyclization, and condensation to form highly toxic advanced glycation end-products (AGEs). These processes are accompanied by oxidative stress, secondary structural perturbations, and altered morphology, progressing toward amyloidogenesis. Metformin, a biguanide, is the most common drug used to treat type 2 Diabetes Mellitus (T2DM).

The aim of this study was to evaluate the protective effect of metformin against fructosylation-induced cross-β structures and amyloid aggregations of human insulin.

UV-absorbance and fluorescence spectroscopy, determination of carbonyl content, free lysine and arginine residues, determination of fructosamine content, SDS-PAGE, circular dichroism (CD) spectroscopy, dynamic light scattering, and scanning and transmission electron microscopy.

Physicochemical studies in the presence or absence of metformin revealed a concentration-dependent structural restoration of fructosylated insulin. Results from the thioflavin-T fluorescence assay suggested that metformin limited the transition of insulin from native to fibrillar state, which was validated by scanning and transmission electron microscopy. Metformin lowered the ThT fluorescence intensity in a concentration-dependent manner. The ThT-specific fluorescence intensity was reduced to 114 and 112.5%. The fluorescence intensity at 2.5 mM metformin was close to native insulin. Electron microscopy revealed that insulin fructosylated by 25 mM fructose in the presence of 2.5 mM metformin suppressed the formation of fibrillar structures. Dynamic light scattering data revealed the potential of metformin to conserve and reinstate the increased hydrodynamic radii (Rh) of fructosylated insulin close to the native conformer. The Rh values of native, fructosylated insulin and insulin incubated with fructose and metformin were found to be 2.65 ± 0.28, 307.6 ± 24.19 nm, and 110.1 ± 4.08 nm, respectively. This study also identified metformin as an antioxidant by protecting critical amino acid residues of the insulin domain.

The study reports the protective effects of metformin on insulin structure, conformation, and function. The findings suggest a potential role for metformin in improving the risk profile associated with insulin resistance due to altered structure or the accumulation of protein aggregates. Interaction studies between insulin and metformin presented here are due to the chemical effect; hence, further in-depth studies are required to identify the molecular mechanism of insulin sensitivity and changes in cellular processes and pathways.

The results suggest that metformin safeguards against fructosylation-induced structural, conformational, morphological, and amyloidogenic aggregating tendencies of insulin. Protein aggregation has been linked to several neurological and metabolic diseases. Hence, metformin may be crucial in preserving the biological activity of insulin by maintaining and protecting its structural integrity and minimizing the associated comorbidities. The study may further be extended to identify the role of metformin in controlling the gradual insulin resistance in T2DM at the molecular level.