Current Medicinal Chemistry - Online First

Centromere protein M (CENPM), a member of the CENP family, is correlated with several malignancies, but its role in colon adenocarcinoma (COAD) is unclear. This study aims to explore the expression, prognostic significance, and biological role of CENPM in COAD.

The association of CENPM with the occurrence and progression of COAD was thoroughly analyzed via several bioinformatics databases. Furthermore, the correlation between CENPM expression and clinicopathological features and prognostic value was validated via immunohistochemistry (IHC) of tissue microarrays (TMAs) from 80 patients.

CENPM mRNA expression was significantly elevated in COAD samples compared with healthy tissues. As COAD progressed, CENPM expression decreased, and patients with lower CENPM transcript levels had a worse prognosis. IHC results further confirmed the overexpression of CENPM in COAD patients, identifying this gene as an independent prognostic factor. Additionally, high CENPM expression was linked to methylation in COAD patients, and the primary function of CENPM and its neighboring genes was determined to be cell cycle regulation. Immunological analysis demonstrated that CENPM expression was positively correlated with activated CD8+ T cells, CD4+ T cells, and dendritic cells (DCs) but negatively correlated with regulatory T cells (Tregs). CENPM expression was positively correlated with that of the immune checkpoint genes LAG3, CD244, LGALS9, PDCD1 (PD1), and PVRL2 but negatively correlated with the expression of BTLA, CSF1R, KDR, IL10RB, PDCD1LG2, and TGFBR1.

These findings collectively highlight a multifaceted role of CENPM in COAD, linking its overexpression to improved patient outcomes through mechanisms involving cell cycle control and immunomodulation. Its significant correlation with key immune infiltrates and checkpoint markers implies potential utility as a novel predictor for immunotherapy responsiveness.

CENPM is an independent prognostic factor for COAD, with its overexpression associated with improved survival. It regulates the cell cycle and tumor microenvironment, making it a promising potential predictive biomarker for immune therapy response.

This study aims to elucidate the role of Phosphoribosyl Transferase Domain Containing 1 (PRTFDC1) in Lung Adenocarcinoma (LUAD) through bioinformatics analysis and experimental validation, exploring its potential as a biomarker for prognosis and treatment response.

We analyzed PRTFDC1 gene expression patterns in 539 LUAD and 59 normal lung tissue samples from The Cancer Genome Atlas (TCGA). Using bioinformatics tools, we examined the correlation between PRTFDC1 expression and clinical characteristics, immune infiltration, Tumor Mutation Burden (TMB), and drug responsiveness. Experimental validation was conducted in LUAD cell lines (A549 and HCC-78) through the overexpression of PRTFDC1, followed by cell proliferation and cell cycle assays.

PRTFDC1 expression was significantly elevated in LUAD compared to normal tissues, correlating with poorer Progression-Free Survival (PFS) and Disease-Specific Survival (DSS). PRTFDC1 was associated with immune cell infiltration, TMB, and mRNA stemness index (mRNAsi). Overexpression of PRTFDC1 in LUAD cell lines promoted cell proliferation and cell cycle progression, mediated by Threonine Tyrosine Kinase (TTK).

The findings suggest that PRTFDC1 may serve as an independent prognostic marker for LUAD, influencing tumor progression and immune response. The correlation with TTK indicates a potential mechanism for PRTFDC1's impact on cell proliferation. However, further research is needed to validate these findings in larger cohorts and explore the underlying molecular mechanisms.

PRTFDC1 is a promising biomarker for LUAD prognosis and treatment response, with potential implications for targeted therapies and personalized medicine.

Diabetic nephropathy (DN) is the main cause of renal failure due to its complexity and difficulty in prevention. The purpose of our study is to screen potential biomarkers of DN at the single-cell level and reveal its new molecular pathogenesis by single-cell RNA sequencing (scRNA-seq).

In this study, scRNA-seq was performed on kidney tissue of control and DN mice. Through multiple analyses of the data, biomarkers in DN that contribute to early diagnosis were screened, and the complex pathogenesis associated with ferroptosis was revealed and verified by experiments at the animal and cellular levels.

Through customized analysis of scRNA-seq results, we found for the first time increased intercellular communication between mesangial epithelial cells and transitional epithelial cells in the pathological state of DN. In addition, two sets of differential protein interaction analysis networks showed that Eno1, Hspa8, FLT1, Hspa1a, and Gsta2 could be used as predictive biomarkers of DN. Finally, the promoting effects of ferroptosis, heat shock protein and their interactions in the development of DN are discussed. In particular, the regulation of GPX4 by members of the heat shock family, Dnaja1 and Hspa1a, promotes lipid peroxidation (the classic phenotype of ferroptosis).

We identified disruption of iron homeostasis and activation of the ferroptosis pathway, alongside differential expression of oxidative stress-related genes, including PGAM2. Heat shock proteins (e.g., Hspa1a, Dnaja1) were found to interact with ferroptosis markers (e.g., GPX4), suggesting a chaperone-mediated protective mechanism under diabetic stress. Analogous to the Flory–Huggins solution theory, HSPs may enhance misfolded protein compatibility in the cytosol, reducing aggregation. This study provides insight into HSP-regulated ferroptosis in DN, though further validation is required for clinical translation.

In conclusion, we comprehensively analyzed the relevant biomarkers and pathogenesis of DN at single-cell resolution, providing new strategies for therapeutic targets of the disease.

The ITIH family, crucial for extracellular matrix stability and cancer progression, is underexplored in multi-omic profiles and immune microenvironments; this study analyzes their roles across cancers and ITIH1’s function in gastric cancer to reveal diagnostic, prognostic, and therapeutic potential.

We analyzed RNA-seq, protein expr ession, and clinical data from 33 cancer types and 24 non-cancerous conditions using TCGA, GTEx, GEO, CPTAC, and IMvigor210 datasets. Methods included differential expression analysis, ROC curve assessment for diagnostic potential, Cox regression and Kaplan-Meier survival analyses for prognostic value, GSEA for pathway enrichment, and molecular docking for ITIH1-targeted small molecule screening. Immune microenvironment interactions, tumor mutational burden (TMB), microsatellite instability (MSI), and immunotherapy response were evaluated. in vitro experiments validated ITIH1’s role in gastric cancer using qRT-PCR, Western blotting, siRNA knockdown, and functional assays.

ITIH family genes exhibited differential expression across cancers and non-cancerous conditions, with ITIH1, ITIH4, and ITIH5 showing high diagnostic potential (AUC > 0.90 in multiple cancers). ITIH1 was a risk factor for poor survival in gastric cancer (p < 0.05). Lower ITIH scores correlated with improved survival in patients receiving immune checkpoint inhibitors (p < 0.05). ITIH genes showed strong correlations with immune checkpoints (PD-1, CTLA-4), TMB, and MSI. Molecular docking identified six small molecules, including Entinostat, with high binding affinity for ITIH1 (-8.4 kcal/mol). ITIH1 knockdown in gastric cancer cell lines (HGC-27, AGS) significantly reduced proliferation, migration, and invasion (p < 0.01).

This study underscores the ITIH family's critical role as diagnostic and prognostic biomarkers across various cancers and non-cancerous conditions, with ITIH1's therapeutic potential in gastric cancer highlighted through its impact on tumor progression, though limitations include discrepancies in some ITIH gene expressions between in vitro and in vivo settings, necessitating further validation.

Our findings highlight the ITIH family's potential as diagnostic biomarkers, prognostic indicators, and therapeutic targets, particularly in gastric cancer. The identification of ITIH1 inhibitors and their association with immune checkpoints, TMB, and MSI paves the way for improved diagnostics, targeted therapies, and immunotherapy predictions, enhancing patient outcomes across diseases.

Cervical cancer, primarily driven by high-risk human papillomavirus (HPV) infection, remains a global health challenge due to limited therapeutic efficacy and adverse effects of conventional treatments. Jinbai Heat-Clearing Prescription (JBHCP), a Traditional Chinese Medicine (TCM), exhibits potential against HPV-associated cervical cancer, yet its molecular mechanisms are unclear. This study aimed to elucidate JBHCP’s multitarget regulatory mechanisms in HPV-induced cervical carcinogenesis.

Network pharmacology, UHPLC-Q-TOF-MS-based metabolomics, and microarray data analysis were integrated to identify the bioactive components and therapeutic targets of JBHCP. Molecular docking and 60 ns Molecular Dynamics (MD) simulations were used to assess the interactions between key compounds (JBHCP673, JBHCP727) and cyclin-dependent kinases (CDK1/CDK2). Gene Ontology (GO), KEGG pathway enrichment, and Protein-Protein Interaction (PPI) network analyses were performed to explore biological functions and signaling pathways.

UHPLC-Q-TOF-MS identified 816 compounds in JBHCP, with 86 meeting drug-likeness criteria. Network analysis revealed 215 shared targets between JBHCP and HPV-induced cervical cancer, including CDK1 and CDK2 as core regulators. Enrichment analysis highlighted JBHCP’s involvement in cell cycle regulation, PI3K/AKT, and STAT3 signaling pathways. Molecular docking demonstrated strong binding affinities of JBHCP727 with CDK1 (-7.36 kcal/mol) and CDK2 (-6.13 kcal/mol). MD simulations confirmed stable binding of JBHCP727 to CDK1/2, while JBHCP673 exhibited instability. ADMET predictions supported JBHCP727’s drug-like properties.

JBHCP exerts anticancer effects by targeting CDK1/2, disrupting cell cycle progression, and modulating oncogenic pathways (PI3K/AKT, STAT3). The stability of JBHCP727-CDK complexes suggests its role in inhibiting HPV-driven proliferation. Multi-component synergy enables JBHCP to act on diverse pathways, aligning with TCM’s “multitarget” paradigm.

This study provides the first systematic evidence of JBHCP’s multi-pathway mechanism against HPV-associated cervical cancer, emphasizing CDK1/2 inhibition as a key therapeutic strategy. JBHCP727 emerges as a promising lead compound. Further in vivo and clinical validation is warranted to translate these findings into clinical applications.

Postoperative hemorrhoidectomy wounds are prone to inflammation and microbial infection due to their anatomical location, necessitating effective therapeutic strategies. CHLoramphenicol (CHL) is a broad-spectrum antibiotic with potential anti-inflammatory properties via Toll-like receptor 4 (TLR4) inhibition. This clinico-computational cohort study investigates CHL’s dual therapeutic mechanism in postoperative hemorrhoid management, combining clinical outcomes with molecular modeling to elucidate its anti-inflammatory and antimicrobial effects.

A prospective, controlled cohort study was conducted with 155 patients (55 CHL, 39 reference treatment [PR], 61 control) undergoing hemorrhoidectomy. CHL ointment (≤120 mg/day) was applied topically until granulation tissue appeared. Clinical outcomes, including edema resolution, granulation tissue formation, and pain scores, were assessed using ImageJ for wound area analysis and the visual analog scale (VAS) for pain. Molecular docking and dynamics simulations were performed using AutoDock and AMBER 22 to evaluate CHL’s binding affinity to TLR4 compared to the reference inhibitor TAK-242. Statistical analyses included ANOVA, Mann-Whitney U tests, and post hoc power calculations.

CHL significantly accelerated wound healing, with 53.2% of patients achieving complete edema resolution by day 3 (vs. 43.6% by day 4) and faster granulation tissue formation (3.58 ± 0.60 days vs. 7.08 ± 1.20 days in control, p<0.0001). Pain scores were significantly reduced in the CHL group. Molecularly, CHL exhibited superior TLR4 binding (ΔGtot = -25.97 kcal/mol vs. -20.69 kcal/mol for TAK-242), with stable complex formation and persistent interactions at Ile-135 (buried surface area: 350 Ų). Healing times were 13.5–19.8 days faster in the CHL group (mean 41 days vs. 54.5–60.8 days in control).

CHL demonstrates dual therapeutic potential in postoperative hemorrhoid management by inhibiting TLR4-mediated inflammation and microbial infection. Its superior binding affinity and clinical efficacy suggest it as a promising multifunctional agent. Further in vitro and long-term studies are needed to validate these findings and explore broader applications in surgical wound care.

Benzoxazole is a privileged scaffold with diverse biological activities, and its hybridization with a 1,2,3-triazole ring can improve affinity and efficacy. This study aimed to synthesize novel 1,2,3-triazole derivatives of 2-aminobenzoxazole and 2-mercaptobenzoxazole, and to evaluate their antiproliferative activity, predicted pharmacokinetic properties, and molecular interactions with kinase targets.

1,2,3-triazole derivatives of 2-aminobenzoxazole 3−15 and 2-mercaptobenzoxazole 18−32 were synthesized via cyclization, propargylation, and copper-catalyzed click reaction. Antiproliferative activity was evaluated against human cancer cell lines: LN-229, Capan-1, HCT-116, NCI-H460, DND-41, HL-60, K-562, and Z-138. The ADME properties of 1,2,3-triazole-benzoxazole hybrids were evaluated using the SwissADME tool. The most active compounds were assessed for Human Gastrointestinal Absorption (HGA) and Blood-Brain Barrier (BBB) permeability using the Egan model. Molecular docking was performed on serine/threonine kinase TAO2 and tyrosine kinase c-Src.

A series of novel 1,2,3-triazole derivatives of 2-amino 3−15 and 2-mercaptobenzoxazole 18−32 were synthesized via click chemistry. Coumarin-containing compounds 3 and 29 showed the most pronounced antiproliferative activity across all tested cell lines. Both demonstrated high predicted HGA and low likelihood of crossing the BBB. Compound 3 exhibited the highest binding affinity for TAO2, while compound 29 showed strong interaction with c-Src.

The results highlight the favorable influence of coumarin substitution on antiproliferative activity, with computational ADME and docking data supporting the observed in vitro efficacy.

This study outlines a viable method for the synthesis of novel 1,2,3-triazole derivatives of 2-aminobenzoxazole and 2-mercaptobenzoxazole. Compounds 3 and 29 demonstrate promising antiproliferative activity and pharmacokinetic potential, supporting their further development as anticancer candidates.

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.