Tumor stromal remodeling is an obstacle for immune checkpoint inhibitors (ICI). A stroma modifying small interfering RNA (siRNA) to carbohydrate sulfotransferase 15 (CHST15) was recently shown to enhance tumor-infiltrating T cells, yet its impact on antitumor response of ICI remains unexplored. In mouse pancreatic cancer KPC and Pan02 subcutaneous syngeneic tumor models, mice were divided into 4 groups for treatment; (1) control, (2) CHST15 siRNA monotherapy, (3) anti-programmed death receptor 1 (PD-1) monotherapy, and (4) combination therapy with CHST15 siRNA and anti-PD-1 antibody. Mice were sacrificed after 2 week-treatments and anti-tumor effects were evaluated by immunohistochemistry for KPC and flow cytometry for Pan02 model, respectively. In the KPC model, combination treatment with intratumoral CHST15 siRNA (0.9-1.0 mg/kg) and systemic anti-PD-1 antibody (5 mg/kg) synergistically and robustly suppressed tumor growth with a significant increase of tumor-infiltrating CD4+ and CD8+ T cells compared to anti-PD-1 monotherapy. In the Pan02 model, combination treatment with CHST15 siRNA and anti-PD-1 showed anti-tumor effect with significant increases in % necrosis area of the tumor, and tumor-infiltrating T cells compared to the control. Notably, the combination therapy dramatically diminishes Ly6C+Ly6G+ granulocytic myeloid-derived suppressor cells (MDSCs) compared to anti-PD-1 monotherapy. The present study demonstrated the robust synergy between systemic anti-PD-1 antibody and a single stroma modifying agent. Combination usage of intratumoral CHST15 siRNA would provide a novel therapeutic option to trigger the remarkable effect of ICI on this most hard-to-treat solid tumor.

Lablab purpureus, a seasonal delicacy in southern India, has been employed in traditional medicinal practices to treat various ailments due to its rich phytochemical content. In this study, the crude methanolic extract of L. purpureus seeds exhibited potent antioxidant, anti-inflammatory, and cytotoxic activities. Antioxidant activity was demonstrated using the DPPH, hydroxyl radical scavenging, ferric oxide reducing and ABTS radical reducing assays, while anti-inflammatory activity was demonstrated using the RBC membrane stabilization, and protein denaturation assays. LPM exhibited significant in vitro cytotoxicity towards MCF-7 and A549 cell lines, causing apoptosis and cell cycle arrests at the S and G2/M phases, respectively, as evaluated using cytometric assays. In addition, the extract exhibited minimal cytotoxicity towards normal HEK-293 cell lines. GC-MS analysis identified the presence of potential compounds in the extract. Commonly overexpressed genes-CDK1, CCNA2, CCNB1, CDC20, and BUB1B-in both the breast and lung cancers were identified from reported microarray datasets. The GC-MS-identified compounds were subjected to molecular docking against the identified proteins using Maestro, Schrodinger (v14.1). Molecular dynamics simulations were performed in an orthorhombic box, with the TIP3P solvent model using Desmond tool of Schrodinger (v14.1) to study the stability of the complexes. Ligands N-Acetyl-D-glucosamine, 4-O-β-D-galactopyranosyl-β-D-Glucopyranose and 2-(acetylamino)-2-deoxy-α-D-Galactopyranose were observed to form stable complexes, in silico with CCNB1, CDC20, and CCNA2 proteins, respectively. Thus Indicating the presence of potent anti-breast and anti-lung cancer agents in Lablab purpureus.

This study employs density functional theory (DFT) and molecular dynamics (MD) simulations to investigate silicon carbide (SiC) nanocrystals as carriers for the anticancer drug Belzutifan. Among tested functional groups (-H, -OH, -NH₂, -COOH), carboxyl-functionalized SiC (SiC-COOH) exhibits superior drug loading capacity with an adsorption energy of -1.03 eV, representing a 25% improvement over conventional carbon-based carriers. The SiC-COOH system demonstrates exceptional stability with a formation energy of -5.42 eV/atom and a remarkably high dipole moment of 142.1 D, facilitating enhanced solubility. Electronic structure analysis reveals significant charge transfers (-0.25e) from Belzutifan to the nanocrystal, accompanied by a substantial reduction in the HOMO-LUMO gap from 5.427 eV (pristine SiC) to 3.41 eV (Belzutifan@SiC-COOH complex), indicating improved electronic coupling. MD simulations confirm the complex's stability under physiological conditions, maintaining structural integrity with root-mean-square deviation (RMSD) values below 2.5 Å throughout the 5 ps simulation. Characteristic shifts in optical spectra (200-600 nm range) and IR vibrational modes (15-40 cm⁻¹) provide clear spectroscopic signatures of successful drug adsorption. The combination of strong binding (-1.03 eV adsorption energy), maintained biocompatibility, and tunable electronic properties positions functionalized SiC nanocrystals as a promising platform for targeted Belzutifan delivery, with potential applications in photo-triggered release systems.

The green synthesis of multifunctional Ag/ZnO nanodots using Artemisia austroyunnanensis leaf extract was explored for their antibacterial, antioxidant, and anticancer properties. Characterization techniques confirmed the successful synthesis of nanodots with a nanoscale size (9.02 ± 1.58 nm), high crystallinity, and distinct ZnO and Ag phases. The synthesized nanodots exhibited significant antibacterial activity against Gram-positive and Gram-negative bacteria, with the largest zone of inhibition observed for Salmonella typhimurium (21 ± 1 mm at 300 µg/mL). Antioxidant assays demonstrated robust free radical scavenging activity, particularly in the ABTS assay (IC50 = 31.85 ± 0.75 µg/mL), attributed to the synergistic effects of ZnO, Ag, and bioactive plant compounds. Cytotoxicity against SHSY5Y neuroblastoma cells revealed dose-dependent activity, with an IC50 value of 193.23 µg/mL, underscoring their potential as anticancer agents. Molecular docking studies highlighted strong binding affinities of Artemisia phytochemicals, particularly artemisinin (- 9.1 kcal/mol), with the p53 tumor suppressor protein, validating their therapeutic potential. These findings demonstrate the efficacy of biogenic Ag/ZnO nanodots as eco-friendly, multifunctional therapeutic agents for bacterial infections and neuroblastoma treatment, offering an innovative approach in sustainable nanomedicine.

In the field of cancer therapy, the diversity and heterogeneity of cancer genomes in clinical patients complicate and challenge the effective use of non-targeted drugs, as these drugs often fail to address specific genetic events. Recent advancements in large-scale in vitro drug screening assays have generated extensive drug testing and genomic data, providing valuable resources to explore the relationship between genomic features and drug responses. In this study, we developed a deep neural network model, DrugS (Drug Response prediction Utilizing Genomic features Screening), utilizing gene expression and drug testing data from human-derived cancer cell lines to predict cellular responses to drugs. Leveraging gene expression and mutation data, we elucidated potential molecular mechanisms underlying SN-38 resistance. Additionally, we used DrugS to evaluate the effects of drugs on cancer cell proliferation in patient-derived xenograft models. In in vitro combination drug experiments, DrugS revealed that CDK inhibitors, mTOR inhibitors, and apoptosis inhibitors effectively reverse Ibrutinib resistance, providing new therapeutic strategies to overcome drug resistance. Furthermore, we assessed the applicability of the DrugS model in drug screening and patient prognosis evaluation using drug information and gene expression data from The Cancer Genome Atlas. In summary, our study offers a novel approach for drug response prediction and mechanism research in cancer therapy from a genomic perspective and demonstrates the potential applications of the DrugS model in personalized therapy and resistance mechanism elucidation.

Bladder cancer (BC) remains a major therapeutic challenge, particularly in patients with acquired resistance to platinum-based chemotherapy. In this study, we investigated the potential of hinokiflavone (HNK), a natural biflavonoid, as a therapeutic agent against cisplatin-resistant BC. Our results demonstrate that HNK differentially inhibited the proliferation of cisplatin-resistant BC cells while sparing normal uroepithelial cells. Mechanistically, HNK induced apoptosis through both intrinsic and extrinsic pathways, as evidenced by caspase activation and Annexin V staining. Next-generation sequencing and gene set enrichment analysis revealed that HNK modulates genes involved in biosynthesis, metabolism, DNA replication and DNA repair. Additionally, HNK downregulated the transcription of MUTYH, OGG1, and XRCC1, which are key genes in base excision repair. For the first time, we identified that HNK as a novel inhibitor of CK2α via in vitro kinase assays, substrate phosphorylation assays, and molecular docking analysis. HNK treatment reduced the phosphorylation of known CK2α targets, including Akt, Stat3, and XRCC1, in cisplatin-resistant BC cells. Time-course analysis revealed that the inhibition of Akt phosphorylation coincided with PARP cleavage, and genetic rescue experiments confirmed the involvement of the CK2α/Akt axis in HNK-induced apoptosis. Furthermore, combination treatment with HNK and chemotherapeutic agents such as doxorubicin or mitomycin C resulted in enhanced cytotoxic effects, suggesting a potential role of HNK as a chemosensitizing agent. HNK, by targeting both DNA repair pathways and CK2-mediated survival signaling, may serve as a promising therapeutic candidate for cisplatin-resistant BC.

Neural stem cells (NSCs) of the ventricular-subventricular zone (V-SVZ) generate diverse cell types including striatal glia during the neonatal period. NSC progeny uncouple stem cell-related mRNA transcripts from being translated during differentiation. We previously demonstrated that Tsc2 inactivation, which occurs in the neurodevelopmental disorder Tuberous Sclerosis Complex (TSC), prevents this from happening. Loss of Tsc2 causes hyperactivation of the protein kinase mechanistic target of rapamycin complex 1 (mTORC1), altered translation, retention of stemness in striatal glia, and the production of misplaced cytomegalic neurons having hypertrophic dendrite arbors. These phenotypes model characteristics of TSC hamartomas called subependymal giant cell astrocytomas (SEGAs). mTORC1 inhibitors called rapamycin analogs (rapalogs) are currently used to treat TSC and to assess the role of mTORC1 in regulating TSC-related phenotypes. Rapalogs are useful for treating SEGAs. However, they require lifelong application, have untoward side effects, and resistance may occur. They also incompletely inhibit mTORC1 and have limited efficacy. Rapalink-1 is a bitopic inhibitor that links rapamycin to a second-generation mTOR ATP competitive inhibitor, MLN0128. Here we explored the effect of Rapalink-1 on a TSC hamartoma model. The model is created by neonatal electroporation of mice having conditional Tsc2 genes. Prolonged Rapalink-1 treatment could be achieved with 1.5 or 3.0 mg/Kg injected intraperitoneally every five days. Rapalink-1 inhibited the mTORC1 pathway, decreased cell size, reduced neuron dendrite arbors, and reduced hamartoma size. In conclusion, these results demonstrate that cellular phenotypes in a TSC SEGA model are reversed by Rapalink-1 which may be useful to resolve TSC brain hamartomas.

BMI1, a constituent of polycomb repressive complex 1, is overexpressed in a variety of cancers, including neuroblastoma, highlighting its potential as a target for cancer therapeutics. Given the pivotal role of BMI1, a number of inhibitors have been synthesized and assessed for therapeutic efficacy across a spectrum of cancers. In our present study, the BMI1 inhibitors PTC-028 and PTC-209 exhibited selective antitumor activity against MYCN-amplified neuroblastoma. Notably, PTC-028, which exhibited toxicity at lower concentrations, triggered apoptosis in neuroblastoma cells and induced G1-phase accumulation, along with reductions in S-phase and G2/M-phase populations, thereby promoting cell cycle arrest. Thorough RNA sequencing analyses revealed that PTC-028 treatment activated the p53 signaling pathway, suggesting it plays a critical role in the mechanism of apoptosis induction. Moreover, PTC-028 treatment led to decreases in levels of anti-apoptotic proteins, including BCL2 and MCL1. Significantly, PTC-028 also exhibited antitumor efficacy in a mouse xenograft model of human neuroblastoma. These results suggest that BMI1 inhibitors, particularly PTC-028, are promising therapeutic agents for the management of aggressive MYCN-amplified neuroblastomas.

The combination of immune checkpoint inhibitors (ICIs), such as ipilimumab and nivolumab, has revolutionized cancer treatment, particularly for advanced melanoma and other solid tumors. However, rare adverse events (AEs) associated with this combination are often underreported in clinical trials. This study aimed to investigate the safety profile of these drugs using the FDA Adverse Event Reporting System (FAERS) database. A retrospective analysis of data from 2015 to the first quarter of 2024 was conducted, focusing on AEs submitted by healthcare professionals. Disproportionality analysis, using algorithms like Reporting Odds Ratio (ROR) and Bayesian Confidence Propagation Neural Network (BCPNN), identified AE signals. A total of 19,462 reports related to the combination therapy were retrieved, spanning 20 System Organ Classes (SOCs), with endocrine and hepatobiliary disorders being most affected. Additionally, 125 unexpected significant AEs were identified, including thyrotoxic crisis, diabetes insipidus, immune-mediated optic neuritis, cystitis, eyelid ptosis, complete atrioventricular block, etc. The highest frequency of AEs occurred within the first 30 days of treatment (38.1%), with a decrease in incidence over time, as indicated by the Weibull Shape Parameter (WSP) test. These findings highlight the importance of continuous monitoring of ICIs in clinical practice.

IWS1 is a key assembly factor of the RNA polymerase II (RNAPII) elongation complex, and its overexpression is associated with worse outcomes in patients with liposarcoma (LPS). This study aimed to identify compounds that can disrupt the IWS1/Spt6 interaction and assess their biological effects in dedifferentiated LPS (DDLPS). Using the AlphaFold-predicted structure of IWS1, we identified a core binding region (AA 545-694) for its interaction with Spt6. Through molecular modeling and virtual screening, Ketotifen and Desloratadine were predicted as candidate inhibitors. Both were predicted to mimic Spt6 phenylalanine (F217) and disrupt the complex, which was confirmed by co-immunoprecipitation. Functional assays showed that treatment with either compound reduced migration, invasion, and spheroid formation in DDLPS cell lines. Additionally, increased nuclear localization of IWS1 was observed. These findings suggest Ketotifen and Desloratadine as promising inhibitors of the IWS1/Spt6 interaction, with potential applications in reducing the invasive properties of human LPS.

Osteosarcoma (OS) is a frequently occurring bone malignancy with increased metastatic properties, causing deaths in large numbers around the world. Disulfiram (DSF) is clinically utilized to treat alcohol dependency and has been indicated to bind Cu(I) in-vivo to form DDTC-Cu(I), which has been confirmed for its antitumor effects. This investigation aimed to elucidate the efficacy of DDTC-Cu(I) on OS cell apoptosis, migration, growth, invasion, and underlying mechanisms. The in-vitro investigations were performed on U2OS, SaOS2, and MG-63 OS cell lines and included CCK-8, colony formation, RTCA, transwell invasion, flow cytometry, wound healing, and RNA seq assays. DDTC-Cu(I) was inoculated dose-dependent, increased apoptosis, and suppressed cells' ability to proliferate, migrate, and invade via the MET and PI3K/AKT signaling pathways. Additionally, MET's overexpression partially reversed the anti-OS and PI3K/AKT signaling pathways suppression effect of DDTC-Cu(I). Furthermore, the SaOS2 xenograft mice model was utilized to confirm the in-vivo anti-OS efficacy of DDTC-Cu(I) by MET protein inhibition. The histological research revealed that DDTC-Cu(I) had no adverse influence on the liver, heart, lungs, and kidneys. Overall, the data of this investigation suggested that DDTC-Cu(I) could serve as an efficient agent against OS development.

This study aims to investigate the clinical characteristics of programmed cell death protein-1 (PD-1) or programmed cell death ligand-1 (PD-L1) inhibitors-induced pituitary injury (pituitary-irAE) by comparing with other pituitary diseases. We retrospectively reviewed clinical records of patients who were admitted for pituitary-irAE, idiopathic isolated ACTH deficiency (IIAD), and lymphocytic hypophysitis (LYH) from January 2015 to June 2024. Among 3689 patients treated with PD-1/PD-L1 inhibitors, 0.596% patients developed pituitary-irAE. The mean age of onset was 59.41 ± 10.69 years, with 13 patients (59.09%) being male. The median duration from symptom onset to confirmed diagnosis was 5.50 (2.00, 9.25) months. The clinical symptoms were similar between pituitary-irAE and IIAD, which mainly manifested as fatigue and appetite loss. Polydipsia/polyuria and amenorrhea/menstrual disorders were mainly found in patients with LYH. Central adrenal insufficiency was observed in all patients with pituitary-irAE, with cortisol concentrations typically falling below the detectable limit. Pituitary MRI appeared normal in 52.63% patients with pituitary-irAE. Furthermore, some patients with pituitary-irAE exhibited partial or complete functional recovery of the pituitary gland after hormone replacement therapy. The clinical manifestations observed in patients with pituitary-irAE are analogous to those seen in individuals with IIAD, yet they differ from the presentations of LYH.

Semi-synthetic derivatives of artemisinin exhibit anti-cancer activity in vitro and in vivo in addition to anti-malarial activity. Here, we report the anti-cancer and anti-cancer stem cell potential of novel C-10 substituted amino-artemisinin derivatives. Of these, the 4'-trifluoromethylarylurea piperazinyl derivative WHN-11 demonstrated cytotoxic activity at high nanomolar concentrations across a range of cancer cell lines. WHN-11 reduced short- and long-term survival of triple-negative breast cancer (TNBC) cells, a highly aggressive breast cancer subtype that currently lacks standardized targeted treatments. Mechanistically, WHN-11 induced a stress response and increased proteasome-mediated turnover of ubiquitinated proteins. WHN-11 promoted mitochondrial depolarization and fission, suppressing the expression of anti-apoptotic B-cell lymphoma extra-large (Bcl-xL) protein and ATP synthesis, thereby decreasing cellular energy production, and inducing apoptosis. WHN-11 treatment also increased autophagosomes, acidic vesicular organelles and lipid droplets. Activation or inhibition of autophagy synergized with the activity of WHN-11 in promoting cellular toxicity, as did increasing cellular dependence on oxidative phosphorylation. Unexpectedly, the effects of WHN-11 appear independent of substantial reactive oxygen species (ROS) production. Taken together, these data suggest that amino-artemisinins related to WHN-11 are promising candidates for anti-TNBC therapies targeting the mitochondria alone or in combination with autophagy modulators.

Echinops L. is recognized for its anticancer and immunomodulatory properties. This study is the first to investigate the potential anticancer and immunomodulatory effects of Echinops shakrokii S.A. Ahmad, collected from Kurdistan, Iraq, employing both in vitro and in vivo methods. Solvent extracts (ethyl acetate, aqueous methanol, and aqueous) from the aerial parts of the plant were prepared via maceration and Soxhlet extraction. The MTT assay assessed antiproliferative efficacy against the MCF-7, MDA-MB-231, EMT6/P, HeLa, T47-D, Caco-2, and VERO cell lines, with results compared to doxorubicin as a positive control. Apoptosis induction was evaluated using an ELISA kit. Immune system activation was assessed through assays of lymphocyte proliferation and macrophages' pinocytic and phagocytic activities. LC-MS identified phytochemicals in the three extracts. EMT6/P cancer cells were injected intraperitoneally into Balb/c mice to investigate the anticancer activity of the aqueous methanol extract. Liver and kidney toxicities were also evaluated. Due to the presence of various phytochemical components, including flavonoids, phenolic compounds, alkaloids, and terpenoids, the ethyl acetate and aqueous methanol extracts exhibited significant antiproliferative effects. The aqueous methanol extract demonstrated the highest potency against the T47D cell line (IC50 = 0.50 ± 0.019 mg/ml), while the ethyl acetate extract displayed the maximum activity against the MCF-7 cell line (IC50 = 0.56 ± 0.03 mg/ml). In vivo, the aqueous methanol extract healed 40% of the mice and reduced the average tumor size by 39.29%. There was no significant increase in renal or hepatic toxicity markers at a dose of 675 mg/kg/day, which was selected based on the acute toxicity test. The plant induced apoptosis through caspase-3 activation. Lymphocyte proliferation, macrophage phagocytosis, and pinocytosis were also enhanced. These findings position E. shakrokii as a potential natural anticancer agent with dual apoptotic and immunostimulatory properties. Its mechanisms should be further investigated for their possible use in cancer therapy regimens.

Osteosarcoma (OS) is a highly aggressive bone cancer in children and adolescents with extremely poor prognosis. Gambogic acid (GA) is a natural compound derived from the resin of Garcinia hanburyi trees and has been reported to possess multiple antitumor effects. However, the comprehensive mechanism of GA intervention in OS remains unclear. In this study, network pharmacology and experiment pharmacology were employed to elucidate the mechanism by which GA inhibited OS. Firstly, GA exhibited significant inhibitory effects on OS in vitro and in vivo. Then, the network pharmacological analysis and molecular docking predicted 9 targets involved in necroptosis and apoptosis induced by GA in OS. The qRT-PCR and western blotting verified that GA could increase the apoptosis-related mRNA and protein expression levels, such as cleaved caspase-3, cleaved PARP, indicating that GA could activate apoptosis. Furthermore, proteins related to necroptosis such as RIPK1, RIPK3, and p-RIPK3 were upregulated, while MLKL initially increased and then decreased, implying a dynamic regulation of necroptosis signaling. In summary, our results identified potential targets and pathways of GA to exert anti-osteosarcoma effect, suggesting that GA is a candidate drug for the treatment of osteosarcoma.

Hypoxia in the tumor microenvironment hinders antitumor immunity. Increasing tumor oxygenation may promote T cell infiltration and tumor control by immune checkpoint blockade (ICB). We found that a radiosensitizer, myo-inositol trispyrophosphate (ITPP), caused oxygen unloading from hemoglobin in CT26 and 4T1 tumors as indicated by photoacoustic imaging (PAI). This change in hypoxia detected by PAI was correlated with strong positive correlations with CD8+ and CD4+ FoxP3- effector T cell (Teff), and negative correlations with monocyte frequencies, indicating that ITPP promoted more immunogenic tumor microenvironments in both models. Combination ITPP and ICB improved tumor control and survival in both models. Therefore, imaging ITPP-modulated tumor hypoxia with PAI was related to ICB treatment response in these studies. Future combination immunotherapy regimens may benefit from monitoring hypoxia using molecular imaging with PAI.

Nanoparticles are promising agents in cancer therapy, yet their cytotoxic mechanisms across diverse cell types require insightful investigation. Bismuth selenide (Bi2Se3) nanoparticles (NPs) was prepared via solvothermal route. The X-ray diffraction (XRD) analysis confirmed that the hexagonal structure with space group R-3 m of the prepared material. The Bi₂Se₃ nanoparticles was found to have a crystallite size of approximately 15 nm. The average NP (powder grain) size and crystallite grain size were determined using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) techniques. The study examines the cytotoxicity and mitochondrial impact of Bi₂Se₃ NPs in tongue cancer cells (SCC-25) and gingival fibroblasts. After 24 h of exposure, significant reductions in cell viability were observed in both cell types, with heightened sensitivity in gingival fibroblasts. Our findings suggest that the reduced crystallite grain size enhances cytotoxicity, likely due to increased nanoparticle-cell interaction and mitochondrial disruption, with cell-specific responses indicating varied vulnerability to mitochondrial toxicity. These results emphasize the critical role of NP size and crystallite size, and their cell-selective interactions in developing targeted and safe Bi₂Se₃-based therapies for cancer treatment.

Lung cancer, particularly non-small cell lung cancer (NSCLC), remains a significant challenge in oncology despite advances in targeted and immune-based therapies. NSCLC accounts for approximately 85% of all lung cancer cases, with five-year survival rates ranging from 4 to 17%, depending on disease stage and regional factors. Chemotherapy resistance remains a major hurdle, contributing to poor patient prognosis. This study explores the therapeutic potential of Sd-021, a novel decursinol derivative, compared to its parent compound, decursin, within various NSCLC cell lines. Our results reveal that Sd-021 demonstrates enhanced anticancer activity, highlighted by a more significant reduction in cell viability, increased induction of apoptosis, and more pronounced cell cycle arrest. Notably, Sd-021 shows increased inhibition of the EGFR/STAT3 signaling pathway in EGFR wild-type cell lines, including A549, H460, and H1299 cells. Moreover, in vivo experiments employing a subcutaneous xenograft mouse model reveal that Sd-021 reduces tumor volume with minimal systemic toxicity, as indicated by histopathological assessments revealing reduced tumor proliferation and heightened apoptosis. The minimal toxicity of Sd-021 offers reassurance regarding its safety for potential clinical applications. In conclusion, these findings highlight the promise of Sd-021 as a therapeutic agent against NSCLC.

Fluoroquinolone-resistant Pseudomonas aeruginosa poses a significant global health concern, particularly in healthcare settings. This opportunistic pathogen has developed resistance against multiple classes of antibiotics, rendering infections challenging to treat. The present study focused on identifying quinoline analogs as potential inhibitors of gyrA in fluoroquinolone-resistant P. aeruginosa. Utilizing structural bioinformatics, molecular docking, molecular dynamics (MD) simulations, and MM/PBSA binding energy analyses, the quinoline analog, N-benzylquinoline-8-sulfonamide (M2), emerged as the most promising candidate. Molecular docking revealed M2's better binding affinity to gyrA wild type as well as frequently observed mutants, demonstrated average binding energy of - 8.14 kcal/mol, significantly better than ciprofloxacin (- 7.13 kcal/mol) and levofloxacin (- 6.57 kcal/mol). M2 exhibited a robust inhibition constant of 1.09 µM, surpassing control antibiotics ciprofloxacin (6.11 µM) and levofloxacin (15.34 µM). MD simulations validated the dynamic stability of M2 and gyrA complexes (wild-type and mutant), whereas MM/PBSA analysis confirmed strong binding energetics. Principal Component Analysis (PCA) further validated the stability of these complexes by identifying the global energy minima across conformational landscapes. M2 exhibited enhanced efficacy and stability against resistance-associated mutations compared to the standard antibiotics ciprofloxacin and levofloxacin. These findings underscore M2's potential as a potent therapeutic agent against fluoroquinolone-resistant P. aeruginosa. Further experimental validation is necessary to confirm its efficacy and to translate these computational insights into clinical applications.

To confirm transcatheter arterial chemoembolization (TACE) combined with apatinib for advanced perihilar cholangiocarcinoma (PCC) is effective and safe. A comprehensive treatment plan involving TACE combined with targeted therapy was implemented for the patients with pathologically diagnosed advanced PCC, where TACE was performed every 4-6 weeks to deliver albumin paclitaxel and gemcitabine for a maximum of six times. Oral apatinib was administered in between TACE cycles. The main endpoint of this study was the objective response rate (ORR), and the secondary endpoints were progression free survival (PFS), overall survival (OS), and adverse events. Kaplan-Meier method was used to assess survival risk factors. From November 2019 to October 2020, a total of 41 patients were enrolled with perihilar cholangiocarcinoma who were pathologically diagnosed. All underwent TACE treatment and received at least two treatment cycles. As of October 2022, the median follow-up period of this study was 28.3 months, the ORR of this study reached 56.1% (95% CI 39.7-71.5%); DCR reached 90.2% (95% CI 76.9-97.3%), and the median PFS was 9.7 months (95% CI 7.6-11.8 months), the median OS was 16.5 months (95% CI 13.6-19.3 months). The treatment-related adverse events (AEs) in this study were mild, mainly Grade 1 or 2. Among the most common AEs were bone marrow suppression and hand-foot syndrome, while no patient had Grade 4 AE. Comprehensive treatment combining TACE with apatinib for advanced PCC had favorable therapeutic effects, and no major safety issue was observed in the patients enrolled.