Antimalarial drug resistance poses a critical threat to global malaria control efforts. Despite the continuous development of novel antimalarial compounds, the emergence of drug resistance remains inevitable, highlighting the urgent need for paradigm-shifting therapeutic approaches. Here, we propose an innovative chimeric antigen receptor-macrophage (CAR-M) cell therapy that circumvents traditional small-molecule limitations by harnessing the innate phagocytic capacity of macrophages. This strategy exploits the specific adhesive interactions between Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) and host receptors (CD36, ICAM-1/CD54, EPCR/CD201) to enable targeted recognition and elimination of infected erythrocytes. By engineering macrophages with chimeric antigen receptors directed against PfEMP1-binding domains, we establish a cell-based immunotherapy platform that provides sustained anti-parasitic activity independent of conventional drug susceptibility profiles. This approach represents a fundamental departure from chemical-based interventions by providing a potentially resistance-proof therapeutic modality for drug-resistant malaria.

The continuous spread of mpox disease caused by mpox virus (MPXV) has posed great threat to global public health. The postattachment membrane fusion process of MPXV is mediated by a multimeric protein machinery, termed as entry-fusion complex (EFC). Among EFC components, A30 and H2 are the earliest identified interaction pair and play important roles in virus entry. Here, we determine the crystal structure of MPXV A30/H2 subcomplex via the tandem-fusion strategy, and show that A30 undergoes large conformational rearrangements upon H2 binding. Structural analysis reveals extended intersubunit interface and highly conserved intermolecular interactions. In vitro binding data further clarify key residues and elements involved in the A30/ H2 subcomplex formation. Finally, we show that the H2-A30 fusion protein, superior to A30 ectodomain alone or the ectodomain-mixture of H2+A30, can induce more potent neutralizing-antibody responses which could inhibit viral infection. These data provide valuable information for the understanding of poxvirus EFC assembly and the H2-A30-based immunogen design and optimization.

Liver fibrosis is a dynamic and reversible pathological process underlying chronic liver diseases, characterized by excessive extracellular matrix deposition and progressive hepatic architectural distortion. It acts as a critical precursor to cirrhosis, hepatic decompensation, and hepatocellular carcinoma, imposing a substantial global disease burden. Accumulating evidence indicates that liver fibrosis is a highly plastic process governed by multicellular crosstalk, immune microenvironment remodeling, epigenetic–metabolic coupling, and mechanotransduction. This review outlines core cellular effectors and their heterogeneity revealed by single-cell omics, and highlights key regulatory layers including circadian rhythm, epigenetic imprinting, metabolic reprogramming, and the gut–liver axis, as well as etiology-specific differences in fibrosis progression, reversibility, and therapeutic response. We also summarize advances in non-invasive diagnosis and clinical translation of anti-fibrotic therapies, and discuss key bottlenecks leading to clinical trial failures. A deeper understanding of cell fate regulation and multicellular ecosystem remodeling will facilitate the development of precise strategies to achieve meaningful fibrosis regression and improve long-term clinical outcomes in chronic liver diseases.

Trichomoniasis, caused by Trichomonas vaginalis, is the most prevalent nonviral sexually transmitted infection worldwide. The occurrence of treatment failure with metronidazole (MTZ) and the adverse effects of currently available nitroimidazoles underscore the necessity for novel therapeutic candidates. From a library of synthetic thienopyrimidine derivatives, G-18 was identified as a lead compound. G-18 exhibited potent anti-T. vaginalis activity against the tested T. vaginalis isolate, with a minimum inhibitory concentration (MIC) of 4 μg/mL and a half-maximal inhibitory concentration (IC50) of 1 μg/mL at 24 h, and it significantly reduced trophozoite viability within 2 h. Flow cytometry confirmed a higher proportion of propidium iodide (PI)-positive parasites following G-18 treatment compared to MTZ treatment (51.2% vs. 10.4%). G-18 demonstrated limited cytotoxicity towards mammalian cells under the tested conditions. Scanning and transmission electron microscopy revealed pronounced ultrastructural damage, including surface roughening, membrane disruption, and organelle disorganization. These findings support G-18 as a promising anti-trichomonal lead compound with rapid in vitro activity and a preliminary selectivity profile under the tested conditions.

The World Health Organization (WHO) “World Malaria Report 2025” outlined global and regional malaria morbidity and mortality patterns; assessed progress toward the WHO’s 2016–2030 Global Technical Strategy milestones; and described funding for malaria initiatives and research. It further examined achievements and gaps across prevention, diagnosis, treatment, elimination, and re-establishment interventions; discussed emerging biological threats; and included a dedicated chapter on escalating antimalarial drug resistance challenges. This article highlights the key points of the report and provides a snapshot of the global malaria burden and advancements. In addition, it briefly summarizes China’s efforts and challenges faced in preventing the re-establishment of malaria transmission, and future prospects for the maintenance of malaria-free status.

Background:Malaria remains a fatal global infectious disease, with the erythrocytic stage of Plasmodium falciparum being its main pathogenic phase. Early diagnosis is critical for effective treatment. This study developed and evaluated an artificial intelligence-assisted diagnosis (AI-assisted diagnostic) tool for malaria parasites. Methods:The peripheral blood samples of malaria patients were collected. Thin blood film smear were prepared, stained and examined by microscopic. After manual confirmation and validation with qPCR, the images of infected red blood cells (iRBCs) of P. falciparum were captured. Using a sliding window method, each original image was cropped into 20 small images (518 × 486 pixels). Selected iRBCs were classified, and P. falciparum was detected using the YOLOv3 deep learning-based object detection algorithm. Results:A total of 262 images were tested. The YOLOv3 model detected 358 P. falciparum-containing iRBCs, with a false negative rate of 1.68% (6 missed iRBCs) and false positive rate of 3.91% (14 misreported iRBCs), yielding an overall recognition accuracy of 94.41%. Conclusion:The developed AI-assisted diagnostic tool exhibits robust efficiency and accuracy in Plasmodium falciparum recognition in clinical thin blood smears. It provides a feasible technical support for malaria control in resource-limited settings.

Conventional neoadjuvant chemotherapy provides limited benefit for patients with resectable non-small cell lung cancer (NSCLC). Recently, neoadjuvant chemoimmunotherapy (NCIT) has transformed the perioperative management of NSCLC by priming systemic anti-tumor immunity before surgery, yet it remains ineffective for at least 50% of patients. Through single-cell sequencing analysis of our NCIT cohort, we identify that antigen-presenting cancer-associated fibroblasts (apCAFs) can impede the efficacy of NCIT. Using a custom cancer-associated fibroblast biobank, we uncover that interferon (IFN)-γ stimulates apCAF expansion via the JAK1/2-STAT1-IFI6/27 pathway. Mechanistically, apCAFs significantly contribute to PD-L2 expression in the tumor microenvironment (TME), triggering the accumulation of FOXP1+regulatory T cells (Tregs) through the PD-L2-RGMB axis. Reprogramming apCAFs by inhibiting the IFN-γ pathway or blocking the PD-L2-RGMB axis substantially mitigates apCAFs-mediated FOXP1+Tregs' expansion. In summary, we reveal the role of apCAFs in compromising NCIT efficacy and propose applications for anti-PD-L2/RGMB regimens to synergize with anti-PD1 therapies by targeting apCAFs.

Lyssavirus glycoprotein plays a crucial role in mediating virus entry and serves as the major target for neutralizing antibodies. During membrane fusion, the lyssavirus glycoprotein undergoes a series of low-pH-induced conformational transitions. Here, we report the structures of Ikoma lyssavirus and Mokola lyssavirus glycoproteins, with which we believe that we have trapped the proteins in pre-fusion and post-fusion states respectively. By analyzing the available lyssaviral glycoprotein structures, we present a sequential conformation-transition model, in which two structural elements in the glycoprotein undergo fine-modulated secondary structural transitions, changing the glycoprotein from a bended hairpin conformation to an extended linear conformation. In addition, such conformational change is further facilitated, as observed in our surface plasmon resonance assay, by the pH-regulated interactions between the membrane-proximal region and the pleckstrin homology and the fusion domains. The structural features elucidated in this study will facilitate the design of vaccines and anti-viral drugs against lyssaviruses.

The persistent emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants highlights the need for developing broad-spectrum antiviral agents. Here, we report the identification of two sarbecovirus S2-specific alpaca nanobodies, namely H17 and H145, that effectively neutralize known SARS-CoV-2 variants (including the Omicron subvariants) and other sarbecoviruses (such as SARS-CoV, PANG/GD, WIV1, and HKU3). The two nanobodies recognize a linear epitope (D1139PLQPELDSFKEEL1152) in the upper region of the S2 stem-helix (SH), which is highly conserved among SARS-CoV-2 variants and other sarbecoviruses. The complex structure of the nanobody bound to the epitope SH-peptide reveal that nanobody binding will impede the refolding of S2, effectively neutralizing the virus. Moreover, the nanobodies bind viral S2 in an acidification-insensitive manner, demonstrating their capacity for entry inhibition especially when viruses enter via the endosomal route. Finally, H17 and H145 possess a better taking-action window for virus neutralization, superior to the RBD-targeting nanobodies that exert neutralization by competing against ACE2 binding. Taken together, the results suggest that anti-SH nanobodies H17 and H145 are promising broad-spectrum drug candidates for preventing and treating the pandemic infections by SARS-CoV-2 variants and other sarbecoviruses.

OBJECTIVE:Transfusion-transmissible infections (TTIs) are severe threats to blood safety and public health. A retrospective study of blood donor records from 2015 to 2019 in Shiyan, China, was conducted. METHODS:TTI prevalence was analyzed using ELISA, RT-PCR, and demographic data. Chi-square test and logistic regression were used to analyze incidence trends and associations for HIV, hepatitis B virus (HBV), hepatitis C virus (HCV), and Treponema pallidum (T. pallidum). RESULTS:Of 230,225 donors, 2,453 (1.07 %) were TTI-positive. HIV, HBV, HCV, syphilis rates were 0.08 %, 0.40 %, 0.19 %, 0.40 %. Significant decreases in HBV, HCV, T. pallidum from 2015 to 2019; HIV decreased from 2018 to 2019. The prevalence is higher among women aged 35-60, farmers with T. pallidum and young female students with HIV infection. CONCLUSIONS:Despite the relatively low prevalence of TTIs at present, awareness of TTIs and ongoing blood screening are still needed to ensure blood safety.

Background: SCTV01E is a tetravalent recombinant COVID-19 vaccine authorized for emergency use in China for adults 18 years and older but not for those under 18. Objective: This Phase 2 trial assessed the safety and immunogenicity of SCTV01E in healthy children and adolescents aged 3 to 17 years, to establish immunobridging with that observed in adults from the efficacy pivotal trial (NCT05308576). Methods: Participants were randomly assigned to receive either 30 µg of SCTV01E or a placebo. Primary endpoints were safety and immunogenicity focused on the geometric mean titer (GMT) and seroresponse rate (SRR) of neutralizing antibodies (nAb) against Omicron BA.5. Results: In total, 268 participants (214 SCTV01E vs. 54 placebo) were included in the safety analysis, with 241 participants (191 vs. 50) in the immunogenicity analysis. Overall, 127 (59.3%) participants receiving SCTV01E and 9 (16.7%) receiving a placebo reported adverse events (AEs), most of which were Grade 1 or 2. No serious adverse events (SAEs) or adverse events of special interest (AESIs) were reported. In the immunogenicity bridging analysis, data from 95 youths were compared with data from 188 adults; the geometric mean ratio (GMR) of the titers was 8.78 (95% CI: 6.05–12.74, p < 0.001), with the lower bound of the 95% CI exceeding 0.67. The difference in the SRR was 6.34% (95% CI: 0.93–11.22%) (p = 0.029), and the lower bound of the 95%CI was >−5%, indicating superiority. Conclusions: SCTV01E was found to be safe and well tolerated in children and adolescents, generating a robust immune response against Omicron BA.5. This supports its potential use in younger populations.

Objective:Bloodborne pathogens including hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), and Treponema pallidum pose a significant menace to transfusion medicine and the public health system. Conducted during the unprecedented global COVID-19 pandemic, this retrospective analysis (2020-2024) evaluated blood donors from Shiyan, China, aiming to ensure blood safety amidst the challenges posed by the pandemic. Methods:Cases of transfusion-transmitted infections (TTIs) were analyzed by using ELISA. Infection rates were quantified as stratified percentages based on sociodemographic characteristics of the study population. Chi-square trend tests were performed to examine the variation in trends. The chi-square test was performed to evaluate associations between sociodemographic characteristics (age, gender, and occupation) and the risk of pathogen infection (HBV, HCV, HIV, and T. pallidum). Results:Analysis of 231,733 blood samples from 114,593 donors revealed an overall 0.51% seropositivity for TTIs, with prevalence rates of HBV (0.23%), HCV (0.03%), HIV (0.03%), and syphilis (0.22%). During the COVID-19 pandemic, a declining trend was observed for all infections. The chi-square test disclosed that HBV and T. pallidum infection were higher in the other occupation group of women aged 35-60 years, HCV positivity was more pronounced in men in the same group, and HIV infection was mainly concentrated in the group of men aged 25-44 years. Conclusions:Continuous monitoring of the prevalence of TTIs among blood donors offers valuable information for ensuring blood safety and new directions in the development of blood transfusion systems.

Cerebral malaria (CM), primarily caused by Plasmodium falciparum, is the primary cause of malaria-related fatalities. CM treatment faces significant challenges due to limited therapeutic options and the emergence of antimalarial drug resistance. Dihydroartemisinin (DHA) is the first-line therapeutic agent for malaria. However, it encounters limitations such as poor solubility, inadequate selectivity, and rapid elimination. Here, we introduced a facile and effective approach using zeolite imidazolium framework-8 (ZIF-8) encapsulated with DHA (DHA@ZIF-8) and assessed its therapeutic efficacy in an experimental cerebral malaria model. The DHA@ZIF-8 demonstrated excellent drug-loading capacity, high stability, prolonged drug release, and improved targeted elimination of Plasmodium parasites within infected red blood cells (iRBCs). This minimized damage to brain microvascular endothelial cells (BMECs) and protected organs from injury, increasing the survival time of the infected mice. Compared to free DHA, DHA@ZIF-8 exhibited better antimalarial efficacy and almost no side effects. This study highlights ZIF-8's potential as a reliable, stable, and efficient drug delivery vector for DHA to improve CM therapy.IMPORTANCEFor the treatment of human malaria, artemisinin-based drugs remain the first-line treatment option. However, their utility is constrained by their short half-life in vivo. Consequently, extending the duration for drug efficacy in the body is a critical issue that needs to be addressed. Metal-organic frameworks are a promising choice for drug loading. In the present study, DHA@ZIF-8 and DHA@MOF were constructed and characterized and were assessed in an experimental cerebral malaria model of C57BL/6 N mice induced by Plasmodium berghei ANKA strain. Data show that DHA@ZIF-8 has a worthy therapeutic effect on experimental cerebral malaria. It will offer a new option for human cerebral malaria (HCM) treatment.

Klebsiella pneumoniae infections (KPIs), particularly carbapenem-resistant Klebsiella pneumoniae (CRKP), pose significant challenges in liver transplantation (LT) recipients, with high morbidity and mortality. Guo et al's study highlights risk factors, such as elevated day-one alanine aminotransferase levels and prolonged catheterization, and identifies polymyxin B and ceftazidime/avibactam as effective treatments. However, limitations like the absence of pre-transplant colonization data and host-pathogen interaction insights highlight the need for enhanced strategies. Future directions should include routine CRKP colonization surveillance, immune and genomic profiling, and the development of novel therapeutics. By integrating these approaches, we can improve the prevention, diagnosis, and treatment of KPIs in LT patients.

For vaccine development against SARS-CoV-2, the spike (S) or spike receptor-binding domain (S-RBD) serves as the major antigen. Enhancing the efficiency and streamlining the process for S/S-RBD purification thus hold considerable practical significance. Here, we identify a basic and evolutionarily conserved region within S-RBD and select a nanobody targeting the region for structural-guided modifications. Histidine substitutions are introduced in the nanobody to enable pH-dependent binding to S-RBD. Two candidates, MNb-11 and MNb-14, are found to readily bind to S-RBD at pH 7.5 but lose the binding capacity below pH 5.0. During the chromatographic-purification trials, resins immobilized with MNb-11 or MNb-14 could purify both wild-type and variant S/S-RBD proteins in one step to a high level of purity and homogeneity. In addition, both modified nanobodies show superior stability across multiple binding-elution cycles. Taken together, our study presents a one-step affinity-chromatographic method for purifying the S/S-RBD protein, which should aid in vaccine development and production against SARS-CoV-2.

Abstract Background Malaria remains a serious public health problem worldwide, particularly in Africa. Resistance to antimalarial drugs is an essential issue for malaria control and elimination. Currently, polymerase chain reaction (PCR) combined with Sanger sequencing is regarded as the gold standard for mutation detection. However, this method fails to meet the requirements of point-of-care testing (POCT) because of its time-consuming, expensive instruments and professional dependence. To support this strategy, we developed a novel diagnostic platform that combines recombinase polymerase amplification (RPA) with the Pyrococcus furiosus argonaute (PfAgo) protein and was designed to detect gene mutations related to antimalarial drug resistance. The Pfcrt haplotypes CVMNK and CVIET of chloroquine resistance (CQR) were used as examples and were assessed in this study. Methods By meticulously designing strategies, RPA primers, guide DNAs, and probes were screened, the reaction was optimized, and the resulting parameters were employed to ascertain the genotype of Pfcrt. The recombinant plasmids pUC57/Pfcrt-CVIET and pUC57/Pfcrt-CVMNK were constructed and diluted for sensitivity detection. The pUC57/Pfcrt-CVIET plasmid mixture was added to the pUC57/Pfcrt-CVMNK plasmid mixture in different additions to configure several specific proportions of mixed plasmid mixtures. The RPA-PfAgo platform was used, and the mixed plasmid was detected simultaneously via nest-PCR (nPCR) and Sanger sequencing. The platform was then evaluated on 85 clinical samples and compared with Sanger sequencing. Results The entire process achieves the key mutation Pfcrt-CVMNK/CVIET genotype identification of CQR within 90 min. The platform achieved 1.8 × 104 copies/μL sensitivity and could detect as little as 3% CVIET in mixed plasmids, which is a higher sensitivity than that of Sanger sequencing (5%). Notably, the platform shows 100% concordance with the gold standard method when 85 clinical samples are tested. The sensitivity and specificity were 100% for the 85 clinical samples. Conclusions This study established an RPA-PfAgo platform for genotyping the key mutation Pfcrt-CVMNK/CVIET of CQR. This method can rapidly produce reliable results and avoid the disadvantages of nPCR with sequencing. This approach has the characteristics of a short operation time, low device dependence, and a good match to the POCT strategy, suggesting that the platform can be easily applied locally or on site. Graphical abstract

IntroductionAccurate and rapid diagnosis is crucial for the effective treatment of parasitosis. Traditional etiological methods, especially microscopic examination, are time-consuming, labor-intensive, and prone to false or missed detections. In response to these challenges, this study explores the use of artificial intelligence (AI) for the detection and classification of human parasite eggs through the YOLOv4 deep learning object detection algorithm.MethodsEggs from species such as Ascaris lumbricoides (A. lumbricoides), Trichuris trichiura (T. trichiura), Enterobius vermicularis (E. vermicularis), Ancylostoma duodenale (A. duodenale), Schistosoma japonicum (S. japonicum), Paragonimus westermani (P. westermani), Fasciolopsis buski (F. buski), Clonorchis sinensis (C. sinensis), and Taenia spp. (T. spp.) were collected and prepared as both single species and mixed egg smears. These samples were photographed under a light microscope and analyzed using the YOLO (You Only Look Once) v4 model.ResultsThe model demonstrated high recognition accuracy, achieving 100% for Clonorchis sinensis and Schistosoma japonicum, with slightly lower accuracies for other species such as E. vermicularis (89.31%), F. buski (88.00%), and T. trichiura (84.85%). For mixed helminth eggs, the recognition accuracy rates arrived at Group 1 (98.10, 95.61%), Group 2 (94.86, 93.28 and 91.43%), and Group 3 (93.34 and 75.00%), indicating the platform’s robustness but also highlighting areas for improvement in complex diagnostic scenarios.DiscussionThe results show that this AI-assisted platform significantly reduces reliance on professional expertise while maintaining real-time efficiency and high accuracy, offering a powerful tool for the diagnosis and treatment of parasitosis. With further optimization, such as expanding training datasets and refining recognition algorithms, this AI system could become a key resource in both clinical and public health efforts to combat parasitic infections.

Drug resistance surveillance is a major integral part of malaria control programs. Molecular methods play a pivotal role in drug resistance detection and related molecular research. This study aimed to develop a rapid and accurate detection method for drug resistance of Plasmodium falciparum (P. falciparum). A quantitative real-time PCR (qPCR) assay has been developed that identifies the mutation at locus A256T in the P.falciparum multi-drug resistance(pfmdr1) gene producing amino acid change at position 86. The results of 198 samples detected by qPCR were consistent with nested PCR and sequencing, giving an accuracy of 94.3%. The sensitivity, specificity, positive and negative predictive value of qPCR were 85.7%, 97.6%, 90.0% and 96.4%, respectively. The results of qPCR are basically consistent with the nested PCR, which is expected to replace the nested PCR as a new molecular biological method for drug resistance detection, providing reliable technical support for global malaria prevention and control.

Corona Virus Disease 2019 (COVID-19) is a highly prevalent and potent infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Until now, the world is still endeavoring to develop new ways to diagnose and treat COVID-19. At present, the clinical prevention and treatment of COVID-19 mainly targets the spike protein on the surface of SRAS-CoV-2. However, with the continuous emergence of SARS-CoV-2 Variants of concern (VOC), targeting the spike protein therapy shows a high degree of limitation. The Nucleocapsid Protein (N protein) of SARS-CoV-2 is highly conserved in virus evolution and is involved in the key process of viral infection and assembly. It is the most expressed viral structural protein after SARS-CoV-2 infection in humans and has high immunogenicity. Therefore, N protein as the key factor of virus infection and replication in basic research and clinical application has great potential research value. This article reviews the research progress on the structure and biological function of SARS-CoV-2 N protein, the diagnosis and drug research of targeting N protein, in order to promote researchers’ further understanding of SARS-CoV-2 N protein, and lay a theoretical foundation for the possible outbreak of new and sudden coronavirus infectious diseases in the future.