Jianhua Li

Four-wheel-drive electric vehicles equipped with independently controllable driving, braking, and steering actuators provide additional degrees of freedom for reducing electric-machine and tire-loss energy. This paper presents a real-time longitudinal–steering coordinated torque-vectoring framework for a vehicle driven by four in-wheel motors. A model-predictive active-front-steering controller coordinates the front-wheel steering angle and external yaw moment to reduce steering resistance and lateral tire-slip loss. A reduced inter-axle propulsion problem is analyzed using the Karush–Kuhn–Tucker conditions, and its speed-dependent switching threshold is calibrated offline by particle swarm optimization. Regenerative braking is allocated by an Energy-Optimized Distribution curve subject to ideal-distribution and regulatory constraints. For general positive-torque operation, sequential quadratic programming distributes the four wheel torques by considering motor input power, tire-slip energy, total torque, yaw-moment demand, and actuator limits. Hardware-in-the-loop results under the United States high-acceleration driving cycle and a double-lane-change maneuver show that the proposed strategy reduces energy consumption relative to uniform and tire-utilization-based torque-vectoring strategies.

Neospora caninum is a major cause of abortion in cattle worldwide, leading to substantial economic losses in the livestock industry. As no effective drug or vaccine is currently available, a deeper understanding of the host immune response against N. caninum is essential for developing effective control strategies. The absent in melanoma 2 (AIM2) inflammasome is involved in host defense and regulation of disease pathology, while its role in N. caninum infection remains unclear. This study shows that N. caninum activates the AIM2 inflammasome in wild-type (WT) murine peritoneal macrophage (PMϕs), characterized by increased expression of AIM2, pro-IL-1β, caspase-1 p20, and IL-1β p17, along with elevated IL-1β secretion and cell death rates. Guanylate-binding proteins (GBPs) are involved in regulating AIM2 inflammasome activation. Here, we observed that both GBP2 and GBP5 were upregulated by N. caninum , but only GBP5 overexpression played a functional role, as its overexpression enhanced, whereas its knockdown significantly attenuated AIM2 inflammasome in WT PMϕs, suggesting N. caninum activates the AIM2 inflammasome in a GBP5-dependent manner. To further investigate the role of AIM2 in parasite infection, AIM2 ^{− / −} mice were used. In AIM2 ^{− / −} PMϕs, inflammasome activation was reduced, accompanied by increased parasite proliferation. In vivo, N. caninum -infected AIM2 ^{− / −} mice exhibited higher parasite loads but showed increased survival rates, reduced macrophage recruitment, decreased levels of IFN-γ, and IL-18, and alleviated pathological damage. In summary, our findings demonstrate that the AIM2 inflammasome is activated by N. caninum in a GBP5-dependent manner and plays a dual role by restricting parasite proliferation while exacerbating disease pathology.

Chicken coccidiosis is a parasitic disease caused mainly by Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix, with most cases presenting as mixed infections. Currently, although a subunit vaccine (CoxAbic) targeting Eimeria maxima via maternal immunization is commercially available, no genetically engineered multivalent subunit vaccine exists against mixed infections caused by these four Eimeria species simultaneously. Therefore, we developed a tetravalent recombinant subunit vaccine (designated TEIN) by fusing key antigen genes (TA4, 3-1E, IMP1, NA4) from these four Eimeria species and expressing the construct in Pichia pastoris. A total of 500 chickens were randomly allocated into 25 experimental subgroups (n = 20 each), consisting of five groups (control, challenged, adjuvant, pPIC9K, and TEIN) and five challenge conditions (infection with Eimeria tenella, Eimeria acervulina, Eimeria maxima, Eimeria necatrix, or a mixture of four species). Immunization was performed via leg intramuscular injection at 14 and 21 days of age. At 28 days of age, all chickens except the controls were orally challenged with 1 × 10⁴ sporulated oocysts. Statistical analysis was performed using one-way or two-way ANOVA as appropriate. Results showed that chickens vaccinated with the TEIN subunit vaccine exhibited significantly elevated serum levels of IgY, IL-2, IL-10, and IFN-γ, as well as an increased splenic lymphocyte CD4⁺/CD8⁺ ratio. The anticoccidial indices (ACI) against Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix, and their mixed infection reached 174.82, 174.58, 174.41, 180.61, and 175.95, respectively. Moreover, no significant differences were observed in hematological parameters, serum biochemical markers, or histopathological findings between the vaccinated and control groups. These results demonstrate the vaccine’s potential as a promising candidate for controlling mixed coccidial infections.

Cryptosporidium parvum is a prevalent protozoan pathogen responsible for moderate to severe diarrhea in humans and animals, posing a significant threat to global public health. Host–parasite interactions play a crucial role in the infection process of C. parvum . Cyclophilin 23 (CpCyP23), a secreted protein of C. parvum , is implicated in parasite development and pathogenicity. As a secretory protein, its function often relies on interactions with host cell proteins. However, it remains unclear whether CpCyP23 has host interaction partners and what specific roles such interactions play during infection. In this study, using CpCyP23 as bait, we screened for potential interacting proteins in HCT‐8 cells through pull‐down assays combined with mass spectrometry (MS) analysis. The interaction was subsequently confirmed by co‐immunoprecipitation (Co‐IP) and bimolecular fluorescence complementation (BiFC) assays. Immunofluorescence (IF) was employed to determine subcellular localization. Furthermore, after confirming the altered expression of the interacting protein during parasite infection, we analyzed its impact on parasite infection using small interfering RNA (siRNA) knockdown and overexpression techniques, coupled with parasite infection assays. The results identified the host cell protein Prohibitin‐2 (PHB2) as an interacting partner of CpCyP23. Co‐IP and BiFC assays validated their interaction, and IF revealed their colocalization in the cytoplasm of HCT‐8 cells. The mRNA level of PHB2 was significantly upregulated at 3 and 12 h postinfection (hpi) with C. parvum . Functional experiments further demonstrated that the expression level of PHB2 positively correlates with the infection burden of C. parvum . In summary, this study reveals an interaction between the host cell protein PHB2 and CpCyP23 and identifies PHB2 expression level as a key host factor significantly influencing C. parvum infection. These findings provide experimental evidence and a potential target for elucidating the pathogenic mechanisms of C. parvum and developing novel anti‐infection strategies.

Giardia duodenalis ( G. duodenalis ) represents a prevalent zoonotic protozoan pathogen responsible for diarrheal disorders across human and animal hosts. To date, no licensed effective vaccines have been developed against giardiasis, and the emergence of drug resistance progressively impairs the efficacy of conventional clinical chemotherapy. Extracellular vesicles (EVs) derived from G. duodenalis (GEVs) exert pivotal functions in mediating parasite immune evasion as well as the initiation of host inflammatory cascades; nevertheless, the molecular regulatory circuits underlying GEV biosynthesis and secretion remain poorly defined. Previous bioinformatic predictions implicate Rab1a in vesicular trafficking within G. duodenalis , whereas its concrete biological contribution to GEV secretion has not been experimentally validated. The present work was designed to screen microRNAs (miRNAs) targeting Rab1a and further elucidate their modulatory effects on GEVs biogenesis. GEVs were purified from G. duodenalis WB‐strain trophozoites, and their identity was verified via transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA, modal diameter: 164.7 nm), plus Western blot detection of classic extracellular vesicle biomarker proteins. High‐throughput miRNA sequencing was subsequently conducted to filter candidate miRNAs capable of interacting with the Rab1a 3′ untranslated region (3′UTR), from which miR‐375, miR‐133, and miR‐999 were shortlisted as prospective regulatory molecules. Functional verification revealed that only miR‐375 robustly repressed Rab1a expression at both mRNA and protein abundances. Dual‐luciferase reporter assays further authenticated the direct physical interaction between miR‐375 and the Rab1a 3′UTR. Consistently, ectopic overexpression of miR‐375 resulted in a prominent decline in cellular GEV secretion. In summary, our experimental data corroborate that endogenous G. duodenalis miR‐375 suppresses Rab1a expression at the post‐transcriptional level to constrain downstream GEV release. The newly characterized miR‐375‐Rab1a‐GEVs regulatory axis deepens our mechanistic understanding of post‐transcriptional modulation governing vesicle formation in G. duodenalis .

Babesiosis, a globally significant tick‐borne disease, poses substantial threats to livestock production and public health. Reported cases of human babesiosis in the United States increased from 1742 in 2014 to 3586 in 2023. In livestock, cattle babesiosis causes mortality, reduced meat and milk production, reproductive losses, and substantial control costs, with annual economic losses estimated at hundreds of millions of US dollars in several endemic countries. Rapid and sensitive detection methods are essential for early warning, surveillance, and control of this disease. In this study, we developed a closed‐tube, one‐pot assay for genus‐level detection of Babesia spp. associated with cattle and sheep, based on recombinase polymerase amplification (RPA) coupled with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a. This format effectively minimizes cross‐contamination risks associated with repeated tube opening in conventional assays. A three‐channel signal readout system, including blue‐light fluorescence visualization, ultraviolet (UV) fluorescence visualization, and lateral flow strip (LFS) readout, was integrated to enable flexible endpoint detection under different laboratory and field conditions. The assay targets a conserved region of the Babesia 18S rRNA gene and enables genus‐level detection of Babesia spp. within 40 min at 37°C. The established RPA‐CRISPR/Cas12a platform exhibited high analytical sensitivity, with a limit of detection of 5 copies/μL for recombinant plasmid templates, high analytical specificity against the tested nontarget pathogens, and low equipment dependency. The detection limit of the LFS format reached 50 copies/μL. Field validation using 71 pooled tick samples and 53 clinical blood samples collected from cattle and sheep yielded positive rates of 15.49% and 9.43%, respectively, with 100% concordance between this assay and conventional polymerase chain reaction (PCR) for both specimen types. In conclusion, this one‐pot RPA‐CRISPR/Cas12a detection platform provides a rapid, sensitive, and field‐applicable molecular screening tool for genus‐level detection of Babesia spp. This assay may support early warning and preliminary field monitoring of babesiosis, particularly in resource‐limited settings. However, species‐level confirmation should be performed by sequencing or other species‐specific methods when epidemiological tracing or precise species identification is required.

Background: Toxoplasma gondii ( T. gondii ) is an important apicomplexan parasite that causes zoonotic toxoplasmosis in humans and animals. Acute T. gondii infection leads to systemic immunopathology that may manifest as lung injury or pulmonary embolism. Ferroptosis is an iron-dependent regulated cell death driven by lethal lipid hydroperoxide accumulation. Emerging evidence implicates ferroptosis in infection-related tissue damage; however, the role of ferroptosis in T. gondii -induced lung injury remains to be explored. Methods: Mice were infected with T. gondii to establish a lung injury model. The body weight changes, survival rate, inflammatory cytokines, lung histopathology, and parasite burden were assessed. The key ferroptosis-related indicators involved in antioxidant, iron metabolism, and lipid metabolism pathways were analyzed in lung tissues using techniques such as transmission electron microscopy, western blotting, and immunohistochemistry. Deferiprone (DFP), an oral iron chelator that can inhibit ferroptosis, was used to investigate the potential role of ferroptosis in T. gondii lung injury. Results: T. gondii infection induced lung injury in mice with thickening of alveolar septa and hemorrhage in alveolar spaces, accompanied by iron deposition. Crucially, T. gondii triggered ferroptosis in lung tissues of mice, evidenced by MDA elevation, GSH depletion, total iron and Fe ²⁺ overload, and mitochondrial cristae loss. Furthermore, iron metabolism pathways were disordered while antioxidant pathways were suppressed. DFP treatment reversed ferroptosis alterations, decreased inflammatory cytokines, attenuated pathological changes, reduced T. gondii burden, and prolonged survival of the infected mice. Conclusions: Our findings revealed that T. gondii infection triggered ferroptosis by compromising dysregulated iron metabolism and antioxidant defenses, playing a key role in T. gondii -induced lung injury. DFP exhibited a promising therapy effect for toxoplasmosis. Graphical