Chloroquine (CQ) resistance has long contributed to antimalarial treatment failure. Despite the implementation of artemisinin-based combination therapies (ACTs) in Gabon, continued surveillance of antimalarial drug resistance markers remains essential for malaria control and elimination. This study aimed to monitor molecular markers of antimalarial drug resistance, specifically the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and P. falciparum multidrug resistance 1 (pfmdr1), in P. falciparum clinical isolates from Libreville, nearly two decades after CQ withdrawal. Archived dried blood spots collected in 2019 were retrieved to assess the prevalence of pfcrt and pfmdr1 mutations and haplotypes in P. falciparum isolates. In total, 70 samples were successfully genotyped via Sanger sequencing. Single nucleotide polymorphisms (SNPs) associated with antimalaria drug resistance were identified in 25.71% of pfcrt and 20% of pfmdr1. For pfcrt, the most frequent mutation was N75(E/K) (24.3%), followed by M74I (22.86%) and K76T (17.14%). Moreover, malaria-positive individuals exhibited mainly the wild-type haplotype CVMNK (74.29%), while the mutant haplotype CVIET (17.13%) was the most common among the mutant variants. The frequency of CVIET observed in this study is substantially lower than the previously reported frequency in Gabon. Other mutant haplotypes, including CVINK, CVMEK, CVMNT, CVIEK, and CVIKK, were each detected at low frequencies (1.43%). Similarly, for pfmdr1, the most frequent mutation was Y184F (18.57%), which predominantly produced the mutant haplotype NEFND (15.71%), compared with YEFND (2.86%) and NEYDD (1.43%). Nevertheless, the wild-type haplotype NEYND was observed in 80% of the study population. These finding indicate the persistence of mutant genotypes in both pfcrt and pfmdr1, along with a decline in their frequencies over time, suggesting a progressive restoration of CQ susceptibility. The results underscore the importance of longitudinal CQ resistance monitoring to accurately assess its prevalence and inform public health interventions.

Lupus nephritis (LN) is characterized by immune system attacks on self-tissues, triggered by the production of pathological autoantibodies, leading to damage in multiple organs and tissues throughout the body. The second-generation calcineurin inhibitor, voclosporin, has recently been recommended in guidelines as an adjunct to basic immunosuppressive interventions for managing active LN in adult patients. However, there are still no real-world data showing that remission was achieved by administering voclosporin in LN using the same protocol as the AURORA trial. We report the case of a 19-year-old woman with proteinuria and microscopic hematuria. The patient developed nephrotic syndrome three years prior to presentation to our department and achieved complete remission with corticosteroids. In the year of presentation, hypocomplementemia and anti-ds-DNA antibodies were detected. Renal biopsy revealed a thickened glomerular basement membrane with spike formation, granular C1q deposition in the mesangial regions, and high electron-density deposits on the epithelial side of the basement membrane. The patient was diagnosed with LN (ISN V) and started treatment with voclosporin combined with corticosteroids, hydroxychloroquine, and mycophenolate mofetil to decrease corticosteroids, which was similar to the regimen as in the AURORA study. Complete remission was achieved 30 days after treatment initiation, and the corticosteroids were discontinued 25 weeks after treatment initiation. This study represents the first clinical case report demonstrating complete remission of LN achieved by implementing the AURORA regimen.

Glioblastoma is an aggressive astrocytic neoplasm characterized by significant intratumoral heterogeneity and resistance to standard therapies. Despite advances such as the Stupp protocol, the prognosis remains poor, with a median survival of 12-15 months. Mast cells in the tumor microenvironment (TME) release pro-inflammatory mediators, promoting tumor progression and therapeutic resistance. The Mas-related G protein-coupled receptor X2 (MRGPRX2), expressed on mast cells, is implicated in neuroinflammatory regulation. This study evaluates hydroxychloroquine (HCQ), a 4-aminoquinoline derivative, as a potential MRGPRX2 inhibitor using a comprehensive multidisciplinary approach. Density Functional Theory (DFT) at the B3LYP/6-31 + G(d,p) level was employed to analyze the structural and electronic properties of HCQ, validated by FTIR and Raman spectroscopy. To ensure statistical robustness and address receptor specificity, 100 independent blind docking simulations were performed using the high-resolution MRGPRX2 structure (PDB ID: 7S8L). The docking protocol was rigorously validated through a cross-docking study using the 7S8N reference ligand, yielding a high spatial consensus with an RMSD of 3.06 Å. The results revealed that HCQ possesses a superior binding affinity of -7.0 kcal/mol, while comparative docking with the standard therapeutic Temozolomide (TMZ) yielded a lower affinity of -5.6 kcal/mol. Detailed residue-level analysis highlighted a distinct binding fingerprint for HCQ, involving specific interactions with residues such as Tyr137, Ser130, and Phe64, which were notably different from the TMZ-receptor complex. ADMET profiling confirmed HCQ's favorable pharmacokinetic properties and its potential to cross the blood-brain barrier. In vitro validation on U87-MG glioblastoma cells demonstrated a time-dependent cytotoxic effect, where HCQ significantly reduced cell viability to approximately 70% at 72 hours (p < 0.01). This biological activity directly correlates with the strong binding profile and high affinity observed in the 100-run docking analysis. Our findings suggest that HCQ is predicted to interact with MRGPRX2 with a high binding affinity, supporting further functional validation and providing a novel structural basis for targeting mast cell-mediated neuroinflammation in glioblastoma treatment. These results advocate for the repositioning of HCQ as a targeted adjuvant therapy to overcome conventional treatment limitations.

Castration-resistant prostate cancer (CRPC) remains lethal due to adaptive resistance mechanisms such as stress-induced autophagy and NF-κB survival signaling. Here, an injectable fiber-in-hydrogel depot is developed for sequential delivery of a multi-enzyme nanozyme (cobalt-epigallocatechin gallate coordination nanozyme, CoNZ) and the lysosomal inhibitor chloroquine (CQ). The preferentially released CoNZ catalytically generates reactive oxygen species, including hydroxyl radicals, and oxygen in situ, inflicting oxidative damage while relieving tumor hypoxia. Simultaneously, it depletes antioxidants (glutathione, NADPH) and impedes NF-κB nuclear translocation, priming CRPC cells for apoptosis. The subsequently released CQ impedes enzymatic degradation or cleavage in endolysosomes and autolysosomes and blocks TLR9/NF-κB signaling, preventing tumor cells from repairing damage or activating pro-survival pathways. In vitro and in vivo, this two-pronged approach synergistically overcomes CRPC's defenses, achieving markedly enhanced cancer cell apoptosis and ∼80% tumor suppression (with occasional complete regression), resulting in an approximately 10-fold reduction in final tumor volume compared to the untreated control, without systemic toxicity, far surpassing single or co-administered treatments. This work demonstrates a spatiotemporally orchestrated combination of nanocatalytic therapy and lysosomal inhibition that dismantles CRPC's resistance mechanisms, highlighting a broadly applicable paradigm for overcoming therapeutic resistance in aggressive cancers.

BackgroundThe commonality of Alzheimer's disease (AD) in the elderly suggests connections between aging and AD biology. APOE biology is also tied to AD.ObjectiveWe sought to link three aging hallmarks (loss of proteostasis, mitochondrial dysfunction, deregulated nutrient sensing) to APOE biology.MethodsWe altered SH-SY5Y cell proteostasis directly via heat shock, integrated stress response inhibition (ISRIB), or autophagy inhibition (chloroquine), and indirectly by perturbing mitochondria (mtDNA depletion; oligomycin). We also exposed induced pluripotent stem cell-derived neurons to ISRIB and chloroquine. Conversely, we mitigated protein stress with rapamycin. We assessed intervention impact on APOE expression.ResultsIncreasing protein stress elevated and decreasing protein stress lowered APOE expression. In SH-SY5Y cells rapamycin blocked oligomycin-induced mTOR 2448 phosphorylation, Akt 473 phosphorylation, and APOE expression. In chloroquine-treated neurons rapamycin reduced mTOR phosphorylation and APOE expression.DiscussionProtein stress initiates APOE expression and facilitates mitochondrial dysfunction's impact on APOE by engaging the mTOR pathway. Our findings link aging and AD biology.

Malaria remains a major health challenge in Angola, where Plasmodium falciparum accounts for most cases and deaths. Although artemisinin-based combination therapies (ACTs) have been the recommended first-line treatments in the country since 2006, the emergence of resistance-associated mutations in Africa raises concerns. We conducted a systematic review of molecular markers of antimalarial drug resistance in Angola (2000-2024) to map their prevalence and distribution at the provincial level. Following PRISMA 2020 guidelines, we searched PubMed, Scopus and Web of Science for studies on P. falciparum resistance markers: polymorphisms in pfcrt, pfdhfr, pfdhps, pfmdr1, pfk13, or copy number variation (CNV) in pfmdr1 and pfpm2/3. Data on mutations, prevalence, and geographic distribution was extracted and summarized. 20 studies met our inclusion criteria. The chloroquine-resistant pfcrt CVIET haplotype persisted in northern and coastal provinces, while the SP-resistant pfdhfr/pfdhps IRNGE haplotype was detected in Cabinda and Zaire. No validated pfk13 mutations linked to artemisinin resistance were reported. Pfmdr1 analyses showed declining 86Y and 1246Y mutation frequencies, with wild-type NYD and NFD haplotypes predominating. Low pfmdr1 and pfpm2 CNV prevalence was reported, though recent data indicate emerging amplification in southern Angola. Research was concentrated in Benguela, Luanda, Lunda Sul, and Zaire, leaving large geographic gaps country-wise. Resistance markers in Angola show heterogeneous provincial distribution. Expanded molecular surveillance, particularly in underrepresented provinces and among pregnant women, is critical to safeguard ACT efficacy and guide national malaria control strategies.

Cervical cancer remains a lethal gynecologic malignancy requiring novel therapies. Here, we investigated the antitumor mechanism of Paris polyphylla saponin Ⅱ (PPSⅡ) in vitro and in vivo. PPSⅡ suppressed proliferation and induced cell death in SiHa, U14, and HeLa cells. The ferroptosis inhibitor ferrostatin-1 (Fer-1) and the autophagy inhibitor chloroquine (CQ) both partially reversed PPSⅡ-induced cell death, indicating involvement of both pathways. Mechanistically, SLC7A11 overexpression reversed the effects of PPSⅡ on GPX4 and ACSL4 expression, whereas BECLIN1 knockdown altered SLC7A11 expression, suggesting that PPSⅡ acts via the BECLIN1/SLC7A11 axis to induce autophagy-dependent ferroptosis. In a HeLa xenograft mouse model, PPSⅡ (1 or 2 mg/kg) significantly reduced tumor volume and weight. Immunohistochemistry showed decreased Ki-67 positivity, and H&E staining revealed tumor necrosis comparable to cisplatin. Notably, PPSⅡ caused no significant pathological changes in heart, liver, spleen, lungs, or kidneys, whereas cisplatin induced renal tubular vacuolation and hepatic inflammation, indicating a better safety profile. In conclusion, PPSⅡ induces autophagy-dependent ferroptosis in cervical cancer cells by modulating the BECLIN1/SLC7A11 pathway and shows potent in vivo antitumor efficacy with low toxicity. These results support PPSⅡ as a promising candidate for cervical cancer therapy.

Mutations in Fms-like tyrosine kinase 3 (FLT3) are strongly associated with relapse and resistance in acute myeloid leukemia (AML) patients, and the treatment of relapsed or refractory AML (R/R AML) remains a major clinical challenge. We previously conducted a prospective clinical trial on R/R AML with chemotherapy regimen BHA (bortezomib, homoharringtonine and cytarabine), which demonstrated promising efficacy in patients with FLT3-mutated R/R AML. However, the therapeutic mechanism remains unclear. In this study, we aim to elucidate the therapeutic mechanism of BHA regimen on the basis of its efficacy for FLT3-mutated R/R AML. We retrospectively analyzed twenty-nine patients with R/R AML, after one course of therapy, patients harboring FLT3 mutations had a significantly higher complete remission/complete remission with incomplete hematologic recovery rate than those without FLT3 mutations (46.67% vs. 7.14%, respectively; P = 0.035). To further explore the underlying mechanisms, we conducted combination index analysis, inhibition of proliferation and apoptosis assays. Compared with 293 T-FLT3 cells, 293 T-FLT3-ITD cells were more sensitive to bortezomib, with significantly lower IC50 values. Bortezomib in combination with homoharringtonine had a synergistic effect on FLT3-ITD cells. Moreover, compared with monotherapy, the combination of bortezomib (4 nM) and homoharringtonine (1 nM) markedly increased total cell death in FLT3-ITD cell lines (MV4-11 and Molm-13). Mechanistically, bortezomib promoted the degradation of FLT3-ITD protein, and the degradation of FLT3-ITD protein was further enhanced by homoharringtonine. Notably, this degradation effect was partially reversed by chloroquine. These findings demonstrate that bortezomib and homoharringtonine have synergistic effects and lead to degradation of FLT3-ITD oncoprotein, potentially contributing to a higher complete remission rate in FLT3-ITD R/R AML.

Intracerebral hemorrhage (ICH) is a neurological disorder characterized by a high mortality rate for which there is currently no definitive cure. Research has demonstrated that adipose-derived mesenchymal stem cells (ASCs) exhibit considerable potential in treating ICH. However, the advanced age of ICH patients and the necessary cell expansion before transplantation therapy could result in the senescence of ASCs, thereby compromising their viability and therapeutic efficacy. This study aims to investigate whether FGF21 (fibroblast growth factor 21) can rejuvenate aged ASCs by enhancing macroautophagy/autophagy flux and subsequently enhance the therapeutic efficacy of ICH. We demonstrated that the autophagy flux of aged ASCs was significantly decreased and FGF21 treatment significantly reversed the senescence phenotype and increased the viability of aged ASCs. Mechanistically, our findings suggested that FGF21 rejuvenates aged ASCs by augmenting autophagy flux, a process partly mediated by TFE3 (transcription factor E3) nuclear translocation. The FGF21-induced TFE3 nuclear translocation was partially facilitated potentially via the FGFR1-SIRT1-MTOR pathway. In addition, FGF21 enhanced the potential of senescent ASCs to differentiate into neurons. In the in vivo study, we further verified that FGF21 could enhance the therapeutic effect of ASCs on acute ICH rats. In conclusion, these results indicated that FGF21 could restore ASC viability by upregulating TFE3-mediated autophagy flux in part through the FGFR1-SIRT1-MTOR signaling pathway, enhanced the potential to improve the differentiation of ASCs into neural stem cells and enhanced the therapeutic effect of ASCs transplantation in acute ICH.Abbreviations: FGF21: fibroblast growth factor 21; TFE3: transcription factor E3; TFEB: transcription factor EB; DMEM: Dulbecco's modified Eagle medium; RAPA: rapamycin; 3-MA: 3-methyladenine; CQ: chloroquine; DMSO: dimethyl sulfoxide; RT-qPCR: quantitative real-time PCR; pAb: polyclonal antibody; mAb: monoclonal antibody; LAMP1: lysosomal associated membrane protein 1; SQSTM1/p62: sequestosome 1; MAP1lc3/LC3: microtubule associated protein 1 light chain 3; GFAP: glial fibrillary acidic protein; MAP2: microtubule associated protein 2; SOX2: SRY-box transcription factor 2; MOI: multiplicity of infection; FGFR1: fibroblast growth factor receptor 1; SIRT1: sirtuin 1; MTOR: mechanistic target of rapamycin kinase; ROS: reactive oxygen species; siRNA: small interfering RNA; OD: optical density; SASP: senescence-related secretion phenotype; IL6: interleukin 6; IL1B/IL-1β: interleukin 1 beta; TNF/TNF-α: tumor necrosis factor; CCL2/MCP-1: C-C motif chemokine ligand 2; BDNF: brain derived neurotrophic factor; VEGF: vascular endothelial growth factor; ICH: intracerebral hemorrhage; MLPT: modified limb placement test.

The aim of present work was to develop and optimise hyaluronic acid-functionalised, chloroquine-loaded solid lipid nanoparticles (HA-CQL-SLNs) for CD44 receptor-mediated therapy of rheumatoid arthritis. The Chloroquine base was precipitated and characterised using UV spectroscopy, FTIR, and DSC. Solid lipid nanoparticles were prepared by melt emulsification followed by probe sonication and optimised using a Box-Behnken design. Particle size, polydispersity index, zeta potential, and entrapment efficiency were evaluated. Ex vivo intestinal uptake was assessed using endocytosis inhibitors, while in vivo pharmacokinetic studies with cycloheximide pretreatment evaluated lymphatic transport. Therapeutic efficacy was studied in arthritic rats. The optimised HA-CQL-SLNs showed a particle size of 412.8 ± 0.71 nm, zeta potential of -9.6 ± 2.3 mV, and entrapment efficiency of 94.6 ± 0.49% (w/w). The lymphatic uptake involved clathrin- and caveolae-mediated endocytosis which was established by reduced systemic drug exposure in animals receiving Cycloheximide pre-treatment. HA-CQL-SLNs significantly reduced paw oedema and joint damage. Thus it can be concluded that HA-CQL-SLNs enhanced bioavailability and therapeutic efficacy, demonstrating potential as a targeted oral delivery system for rheumatoid arthritis.

Enteroviruses pose a substantial global health burden, and a complete understanding of host defense mechanisms is still evolving. Through proteomic profiling of coxsackievirus B3 (CVB3)-infected cells, we identified the host factor USF2 (upstream transcription factor 2) as significantly upregulated, a process that may be associated with the viral capsid protein VP4. Here, we characterize USF2 as a novel, infection-induced restriction factor. Functional studies demonstrate that USF2 knockdown enhances viral RNA replication without affecting entry, while its overexpression suppresses replication and cytopathic effects. Mechanistically, USF2 acts as a transcriptional repressor of autophagy-related genes, thereby inhibiting autophagosome formation. Crucially, USF2 depletion promotes the release of MAP1LC3/LC3-positive extracellular vesicles carrying infectious virus, defining its role in blocking a key route of vesicle-mediated viral dissemination. USF2 exhibits broad antiviral activity against multiple enteroviruses and other RNA viruses that depend on cytoplasmic membrane remodeling. Our findings reveal that the host deploys a virus-triggered transcriptional regulator to restrict enterovirus spread by suppressing macroautophagy/autophagy-dependent viral dissemination.Abbreviations: BafA1: bafilomycin A1; CQ: chloroquine; CPE: cytopathic effect; CVB3: coxsackievirus B3; dpi: days post infection; DEGs: differentially expressed genes; EV-A71: enterovirus A71; EVs: extracellular vesicles; GFP: green fluorescent protein; LAMP1: lysosomal-associated membrane protein 1; TR: LysoTracker Red; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MOI: multiplicity of infection; TFEB: transcription factor EB; USF2: upstream transcription factor 2; UPR: unfolded protein response; VP1: viral capsid protein 1; VP4: viral capsid protein 4.