Among hematologic immune-related adverse events (irAEs), autoimmune hemolytic anemia (AIHA) is rare and often fatal. We report a fatal case of irAE-associated AIHA that developed more than six months after the first administration of pembrolizumab.The patient was a 70-year-old man who was referred to our department after a thickened pleural lesion was detected on chest radiography performed during follow-up for trigeminal neuralgia. Pathological examination of a biopsy specimen revealed clear cell renal cell carcinoma, and the pleural lesion was diagnosed as a metastasis from renal cell carcinoma. Contrast-enhanced CT demonstrated a tumor extending into the renal vein, para-aortic lymph node metastasis, and right pleural metastasis, leading to a diagnosis of metastatic renal cell carcinoma, cT3aN1M1. Combination therapy with pembrolizumab and lenvatinib was initiated as first-line treatment. After seven courses over approximately six months, disease progression was observed, and cabozantinib was introduced as second-line therapy.About one month after starting cabozantinib, anemia developed, followed by dizziness, prompting emergency hospitalization. Rapid progression of anemia was observed, and the patient was admitted to our hospital. AIHA caused by pembrolizumab-related irAE was suspected, and high-dose corticosteroid pulse therapy was initiated. Although transient improvement was achieved, the anemia acutely worsened again, resulting in cardiac arrest. A total of 14 units of blood transfusion stabilized circulation; however, renal dysfunction and consciousness disturbance secondary to cardiac arrest did not improve, and the patient ultimately died of renal failure.We report this case with a review of the literature, discussing the clinical course and management challenges of AIHA as a delayed hematologic irAE following immune checkpoint inhibitor therapy.

We report a silver-based metallosupramolecular polymer system designed for multifunctional therapeutic applications, in which metal-organic coordination motifs integrated into a hydrophilic oligomer enable spontaneous self-assembly into stable nanostructures that exert broad-spectrum antibacterial activity, selective anticancer efficacy, and potent antiviral performance. We hypothesized that cytosine-silver-cytosine (Cy-Ag-Cy) coordination could regulate polymer association and nanoscale self-assembly, thereby coupling molecular organization with the colloidal and stimulus-responsive properties of the resulting nanogels. Herein, we construct a water-soluble supramolecular polymer (Ag-CyPEG) featuring cytosine-silver-cytosine coordination units, which self-assemble in aqueous media into nanogels that exhibit strong intrinsic fluorescence, pronounced pH responsiveness, high stability in biological environments, and excellent anti-hemolytic behavior, indicating suitability for biomedical applications. Biofunctionally, Ag-CyPEG nanogels exhibited excellent antibacterial activity against Gram-positive and Gram-negative bacteria, including drug-resistant strains, achieving efficient cell killing with rapid kinetics. The nanogels also showed strong anticancer selectivity, as they induced substantial apoptosis across multiple cancer cell lines but exerted negligible toxicity toward normal cells, attributed to selective cellular internalization and acid-triggered disassembly within cancer cells. Importantly, the nanogels also suppressed H1N1 influenza A viral activity prior to cellular entry and prevented viral propagation, indicating their ability to suppress infection. Collectively, these findings establish a coordination-driven nanogel platform in which molecular Cy-Ag-Cy interactions are translated into nanoscale self-assembly, colloidal properties, and stimulus-responsive behavior, providing a conceptual framework for the design of multifunctional metallosupramolecular materials with integrated biological functions. SIGNIFICANCE OF THE STUDY: This study establishes a generalizable metallosupramolecular design strategy that unifies intrinsic fluorescence and broad-spectrum bioactivity within a single silver-cytosine coordination-driven nanoplatform. Beyond integrating antibacterial, anticancer, and antiviral activities, this system defines a mechanistic framework in which microenvironment-responsive metal-ligand interactions enable selective activation of silver ion-mediated therapeutic action. This strategy shifts nanomedicine design from static, single-function systems toward adaptive, coordination-programmed platforms with context-dependent bioresponsiveness. Importantly, this provides a transferable blueprint for engineering multifunctional supramolecular therapeutics with tunable activity, high biocompatibility, and minimized systemic toxicity. More broadly, this work establishes a new paradigm for precision nanomedicine, in which structural programming at the supramolecular level enables rational development of intelligent, multi-target therapeutic systems with broad translational impact.

Probiotics have been reported to effectively modulate human glucose metabolism. Here, we aim to isolate and identify Lacticaseibacillus paracasei SX52-B (L. paracasei SX52-B) from the intestines of centenarians, investigate its hypoglycemic properties, and explore the potential molecular mechanisms through integration of genomic analysis. To evaluate the probiotic properties, L. paracasei SX52-B was subjected to growth characteristics, safety and functional tests. Whole-genome sequencing of L. paracasei SX52-B was performed to identify functional genes related to hypoglycemic properties. Genomic annotation was conducted by comparing against databases such as GO, COG, KEGG, and CAZy. Firstly, L. paracasei SX52-B exhibited great biosafety across a series of assays. It could survive in simulated gut microbiota juices, indicating significant tolerance to acidic conditions and bile salts. Moreover, L. paracasei SX52-B showed no hemolytic activity and exhibited antibiotic susceptibility within safe limits. In the acute oral toxicity test of mice, neither adverse reactions nor significant differences in organ indices were detected after they were fed different doses of L. paracasei SX52-B for 28 days. Genomic sequencing and analysis revealed that the genome size of SX52-B is 2,991,568 bp. Through functional annotations, relatively rare carbohydrate metabolism-related genes such as malX (maltose/glucose PTS system EIICB component), tpiA (triosephosphate isomerase) and E5.1.3.15 (glucose-6-phosphate 1-epimerase), were identified in L. paracasei SX52-B, which were rarely detected in common L. paracasei strains. These results suggest that L. paracasei SX52-B possesses distinct genomic features related to glucose-lowering pathways. On the other hand, the result of in vitro functional assay demonstrated that L. paracasei SX52-B exhibited a high α-amylase activity inhibition rate (94.67%) and favorable lipid-lowering capacity. The supernatant of L. paracasei SX52-B also exhibited significant antioxidant activity. To verify the hypoglycemic capacity of L. paracasei SX52-B in vivo, we employed a hyperglycemic zebrafish model. It is revealed that both supernatant and bacteria of L. paracasei SX52-B significantly reduced the glucose content in zebrafish (P < 0.001). The study findings highlight that L. paracasei SX52-B exhibits favorable probiotic properties and hypoglycemic capacity, holding promising potential for application in the development of functional hypoglycemic foods or pharmaceutical products.

Cold antibody-mediated autoimmune hemolytic anemia, or cold agglutinin syndrome (CAS), in pediatrics is typically transient, although it can often be severe and require transfusion support. Recurrence in children is rare, but those with underlying immune dysfunction can have a refractory course with limited options available. We report a 4-year-old girl with Evans syndrome and life-threatening CAS refractory to transfusions, plasmapheresis, steroids, and immunosuppression as well as a 12-month-old boy status post liver transplant with CAS refractory to steroids, rituximab, and tacrolimus cessation with transition to sirolimus, both of whom received sutimlimab, a C1s inhibitor, with rapid improvement in hemolysis, transfusion independence, and a sustained remission. To our knowledge, these are the first reported uses of sutimlimab in the pediatric population.

Spondyloenchondrodysplasia (SPENCD) is a rare monogenic interferonopathy caused by biallelic ACP5 mutations, characterized by skeletal dysplasia, neurological involvement and immune dysregulation frequently manifesting as lupus-like disease. Autoimmune cytopenias, particularly autoimmune hemolytic anemia (AIHA), are among the most severe and treatment-refractory manifestations. Anifrolumab, a monoclonal antibody targeting interferon alpha receptor subunit 1 (IFNAR1), blocks signaling by all type I interferons and is approved for moderate-to-severe systemic lupus erythematosus. We report a 50-year-old man with SPENCD-associated lupus and refractory, transfusion-dependent AIHA who achieved sustained hematologic remission following anifrolumab initiation. Using this case as a framework, we discuss the pathophysiological basis of SPENCD, the spectrum and treatment challenges of its associated autoimmune cytopenias, and the evolving role of targeted therapies in monogenic interferonopathies, with particular emphasis on SPENCD, and summarize the published experience with anifrolumab in these disorders. Collectively, this case and the available literature provide preliminary support for IFNAR1 blockade as a mechanism-based, steroid-sparing strategy for refractory autoimmune cytopenias in SPENCD.

The D-- phenotype is an exceptionally rare Rh blood group phenotype characterized by expression of D in the complete absence of C, c, E, and e. Individuals with this phenotype are at risk of developing anti-Rh17 (anti-Hr0), a clinically significant alloantibody that can severely complicate transfusion support and pregnancy management because compatible blood is exceptionally difficult to obtain. This review critically synthesizes the available published evidence regarding the D-- phenotype and anti-Rh17 alloimmunization, integrating data from case reports, case series, family investigations, and molecular studies from different geographic regions. Current evidence regarding molecular mechanisms, immunohematologic character istics, serologic identification, transf usion management, pregnancy outcomes, and rare donor strategies is summarized, w ith emphasis on practical considerations relevant to transfusion medicine. Although the D-- phenotype is exceptionally rare, early recognition, comprehensive immunohematologic evaluation, and proactive transfusion planning, including collaboration with rare donor programs, are essential for optimizing patient outcomes.

Although Immune cytopenias such as immune thrombocytopenia (ITP), autoimmune haemolytic anaemia (AIHA) and pure red blood cell aplasia (PRCA) are well documented associations with chronic lymphocytic leukaemia (CLL), aplastic anaemia (AA) is seemingly rare.

Quinuclidine derivatives are potent bioactive compounds investigated as cholinesterase inhibitors with potential intravenous therapeutic application. However, the cytotoxic properties of some, particularly those linked to oxidative stress, pose significant safety concerns. This study aimed to evaluate if flavonoid fisetin could protect cells from such toxic effects. Erythrocytes were chosen due to high susceptibility to reactive oxygen species (ROS) and well-characterised endogenous antioxidant system, making them a sensitive and physiologically relevant model. Over 24 h, human erythrocytes were exposed to QOH-C12-16 or QNOH-C12-16 derivatives to establish their 25, 50, and 75 % inhibitory concentrations (IC25, IC50, and IC75, respectively). As the compounds with the C12 side alkyl chain showed no significant haemolytic effects, they were excluded from further experimentation. In the next steps, we assessed the protective effects of fisetin (1, 10, 100 μmol/L) against quinuclidine-induced haemolysis over 24 h and against reactive oxygen species (ROS) generation and potential glutathione (GSH) and superoxide dismutase (SOD) depletion over 4 h. Fisetin at 10 and 100 μmol/L significantly reduced ROS to near-control levels, restored SOD activity, and decreased haemolysis by 10-40 %, most effectively in cells exposed to quinuclidines at IC50. However, GSH measurements in combined treatment were not reliable, as fisetin exhibited autofluorescence, but single quinuclidine exposure showed no significant effects on GSH levels. In summary, fisetin showed a promising potential to make quinuclidine intravenous therapies safer.

Anemia, characterized by decreased hemoglobin levels and impaired oxygen-carrying capacity, remains a major global public health concern. Conventional therapies, such as blood transfusions and oral iron supplementation, are often associated with adverse effects and limited absorption, highlighting the need for safer and more effective alternatives. Natural products rich in polyphenols and essential minerals have attracted increasing attention due to their multifunctional therapeutic potential. In this context, the present study is the first to investigate the anti-anemic potential of Pinus pinaster Aiton and Pinus halepensis Mill. bark extracts through an integrated phytochemical, nutritional, in vivo, in vitro, and in silico approach. Phytochemical profiling by HPLC-MS/MS revealed several major phenolic compounds, including quercetin, catechin, protocatechuic acid, and taxifolin-3-O-glucoside. The extracts exhibited high total polyphenol and flavonoid contents and strong antioxidant activity, as demonstrated by DPPH, ABTS, PR, and TAC assays. Mineral analysis revealed significant levels of essential elements, including iron, magnesium, potassium, sodium, and zinc. In phenylhydrazine-induced anemic Wistar rats, 21-day treatment with the extracts significantly restored hematological parameters (RBC, Hb, and HCT), serum iron and ferritin levels, and improved renal and hepatic biomarkers compared with untreated anemic controls. In vitro assays further demonstrated anti-hemolytic activity and enhanced erythrocyte membrane stability. Molecular docking analysis supported these findings by revealing favorable interactions between the major phenolic compounds and key targets involved in erythropoiesis, iron metabolism, and oxidative stress, including the erythropoietin receptor, superoxide dismutase, and ferrochelatase. Collectively, these findings highlight the potent anti-anemic and protective effects of Pinus bark extracts and suggest their potential as promising natural therapeutic agents.

Bacteriophages have been widely recognized as one of the most promising alternatives to antibiotics. In this study, a multivalent bacteriophage designated vB_SalM_Sz was isolated, and its whole-genome sequence, biological traits, biosafety profile, and bactericidal activity were comprehensively characterized. Genomic and phenotypic analyses revealed that vB_SalM_Sz is a polyvalent phage capable of lysing both Salmonella spp. and Escherichia coli, taxonomically classified within the genus Rosemountvirus, class Caudoviricetes, family incertae sedis. Phage vB_SalM_Sz exhibited robust stability under broad environmental conditions, including temperatures ranging from 30 to 50 °C, pH values of 3-10, and NaCl concentrations of 0-3 M. Importantly, this phage exerted no hemolytic toxicity and did not impair the viability of mammalian cells. Furthermore, vB_SalM_Sz displayed dose-dependent growth inhibition and bactericidal activity against single cultures of Salmonella, E. coli, and their mixed co-cultures in LB medium, milk, and chicken matrices. The phage treatment significantly improved survival rates in both G. mellonella and chick infection models, while mitigating intestinal tissue injury and suppressing excessive inflammatory responses triggered by Salmonella strain S43 in infected chicks. Collectively, these findings demonstrate the great application potential of the multivalent phage vB_SalM_Sz for the biocontrol of Salmonella and E. coli, laying a solid theoretical and experimental foundation for its further translational development as an antibacterial agent.

The role of native bacterial probiotics in the Chinese traditional fermentation starter, Qubing, has not been previously studied. In this investigation, 22 bacterial strains were isolated from Qubing, and potential probiotic candidates were selected for their antibiofilm activity against Burkholderia thailandensis and Staphylococcus aureus. The most resistant bacterial isolates were highly tolerant of simulated harsh conditions, including acid, bile salts, and gastric juice. Many isolates exhibited bile-salt hydrolase activity, suggesting that they reduce cholesterol levels by deconjugating bile acids. Furthermore, numerous isolates exhibit greater auto-aggregation and hydrophobicity, enabling adhesion to the gastrointestinal tract and facilitating colonization while preventing pathogen invasion. Moreover, culture-free supernatants (CFSs) from the isolates showed considerable antibacterial effect against B. thailandensis and S. aureus. Tricine-SDS gel electrophoresis revealed protein bands at 8 kDa, indicating bacteriocin-like activity. The proteolytic, amylolytic, lipolytic, and cellulolytic activities were observed in the majority of isolates, whereas a few isolates demonstrated β-galactosidase, phytase, and exopolysaccharide production. Almost all isolates demonstrated notable cholesterol assimilation and free radical-scavenging activities. The absence of antibiotic resistance and the non-hemolytic properties underscore safety. Bacillus subtilis QB-07 and Bacillus albus QB-10 were identified as potential isolates by principal component analysis. The B. albus QB-10 CFS showed a dose-dependent reduction in biofilm formation by the tested bacterial pathogens, confirmed by microscopic observation (CLSM and SEM). RT-qPCR studies exhibited that CFS differentially regulates biofilm-associated virulence gene expression in tested pathogens. Thus, the promising probiotic B. albus CFS could serve as an alternative source for identifying novel antibiofilm peptides in the future.

Among the various proteins found in the venom of snakes belonging to the genus Bothrops, which is responsible for the highest number of snakebite accidents in Brazil, Phospholipase A2 (PLA2) is one of the most important components. The present study demonstrated the isolation of a fraction containing two Asp49 PLA2 isoforms from the endemic southeastern Brazilian snake Bothrops fonsecai, named BfPLA2. The isolation was achieved through two chromatographic techniques: RP-HPLC using a Teknokroma Europa Protein 300 C18 column, followed by anion-exchange chromatography using a HiTrap Q FF column. SDS-PAGE and two-dimensional electrophoresis revealed proteins with an approximate molecular mass of 15 kDa and an isoelectric point close to 5, indicating acidic PLA2 isoforms. Mass spectrometry analysis and bioinformatic analysis identified two isoforms with high similarity to PLA2s previously described in literature, corroborating the protein family of the isolated toxins. BfPLA2 showed maximum activity at 60 °C and pH 8.5 (Mean: 223.89 U/min/mg). Cytotoxic activity against the HeLa cell line was demonstrated (IC50: 26.21 μg/mL), as well as anticoagulant activity, prolonging coagulation time by 74.1% at the highest dose, and hemolytic activity (IC50: 8.26 μg/mL). The characterization of proteins isolated from snake venoms may contribute to a deeper understanding of snakebite envenomation, especially regarding Bothrops fonsecai, whose venom remains poorly characterized.

Invasive Group A Streptococcus (iGAS) infection is a disease state with increasing incidence internationally. This case report outlines a patient with severe septic shock from Group A beta-haemolyticStreptococcus within a provincial intensive care unit (ICU) in New Zealand. Consequently, this case has prompted discussion on the local change in epidemiology for Group A beta-haemolytic Streptococcus (GAS) and a review of the evidence of recognised interventions for iGAS and Streptococcal toxic shock syndrome (STSS).