Wanglong Zheng

CD163 not only acts as an essential receptor for porcine reproductive and respiratory syndrome virus (PRRSV) infection but also serves as a barrier to cross-species cellular infection by arteriviruses. Among its domains, the scavenger receptor cysteine-rich 5 (SRCR5) domain of CD163 is functionally indispensable. In this study, two monoclonal antibodies (mAbs), designated 5A and 11D, were developed against the SRCR4-6 region of porcine CD163. The linear epitopes recognized by mAbs 5A and 11D were identified as ⁵⁴⁶CEGHESHLSLCPVAP⁵⁶⁰ in SRCR5 and ⁴⁴⁹WDCKNW⁴⁵⁴ in SRCR4, respectively. Crystal structural analysis and sequence alignment showed that the ⁵⁴⁶CEGHESHLSLCPVAP⁵⁶⁰ epitope resides in the long loop 5-6 of SRCR5, which is a key structural region responsible for ligand binding. Meanwhile, the ⁴⁴⁹WDCKNW⁴⁵⁴ epitope is located within a highly conserved region of SRCR4, which confers broad cross-species reactivity. Notably, mAb 5A reduced PRRSV infection, while mAb 11D exhibited no anti-PRRSV activity in target cells. Collectively, this study provides essential molecular tools for exploring CD163-domain-dependent cross-species reactivity and developing anti-PRRSV strategies.

Feline and canine coronaviruses (FCoVs and CCoVs) are widely prevalent in companion animals. The recent emergence of a highly virulent FCoV-CCoV recombinant virus, FCoV-23, caused a rapid feline infectious peritonitis (FIP) outbreak in Cyprus, which raises a serious concern about the cross-species transmission of CoVs among companion animals. Here, we evaluated the prevalence, transmission and evolution of FCoV and CCoV in the Jiangsu and Zhejiang provinces of China in 2025. A universal RT-qPCR assay for FCoV and CCoV was established and used for the detection of 700 clinical samples, including 501 from cats and 199 from dogs. A total of 92 samples (13.14%, 92/700) were detected as positive, containing 79 positive samples (15.76%, 79/501) from cats and 13 positive samples (6.5%, 13/199) from dogs. Ten complete S genes and two complete genomes were obtained from positive samples. Fragment comparison and phylogenetic analysis identified one CCoV-I sequence from a feline sample and one FCoV-II sequence from a canine sample. Furthermore, recombination analyses not only detected intra-species recombination events but also inter-species cross-over events. Collectively, this study, for the first time, revealed that cross-species transmission and recombination events occurred between FCoV and CCoV among companion animals in the Jiangsu–Zhejiang region of China in 2025.

Manganese ions (Mn²⁺) are an essential trace element within organisms spanning the entire tree of life. It has reported that Mn²⁺ exerts strong immunocompetence effects and exhibits antiviral effects against various human and animal viruses, including DNA and RNA viruses. Recently, Mn²⁺ has been found to be involved in the activation of the innate immune DNA-sensing cyclic GMP-AMP synthase (cGAS) stimulator of interferon genes (STING) pathway and subsequent antiviral function. However, the antiviral mechanism of Mn²⁺ remains unclear. In the current study, the results suggest that the cGAS-STING pathway is essential for Mn²⁺ to promote interferon (IFN) signaling, but it is not essential for triggering antiviral functions. After knocking out the STING or interferon regulatory factor 3 (IRF3) gene, Mn²⁺ still retains its antiviral activity against herpes simplex virus type 1 (HSV-1) and vesicular stomatitis virus (VSV). Furthermore, the results from transcriptomic analysis indicate that Mn²⁺ can induce a significant change in the apoptotic process in STING⁻/⁻ 3D4/21 cells. Mn²⁺ can induce cell apoptosis through the oxidative stress pathway, and inhibiting the apoptotic signal could suppress Mn²⁺-mediated antiviral activity in STING⁻/⁻ 3D4/21 cells. Additionally, dual knockout of IRF3 and caspase3, resulting in concurrent loss of IFN and apoptotic signals, eliminates the antiviral effects of Mn²⁺. In summary, the current study suggests that Mn²⁺ could exert antiviral effects not only through the cGAS-STING-IFN pathway but also via the reactive oxygen species (ROS)-apoptosis pathway.

Manganese (Mn²⁺) is an essential trace element within organisms spanning the entire tree of life. It has reported that Mn²⁺ exerts strong immunocompetence and exhibits antiviral effects against various human and animal viruses, including DNA and RNA viruses. Recently, Mn²⁺ has recently been found to be involved in the activation of the innate immune DNA-sensing cGAS-STING pathway and subsequent anti-virus function. However, the antiviral mechanism of Mn²⁺ remains unclear. In current study, the results suggested that cGAS-STING pathway is essential for Mn²⁺ promoting the IFN signaling, but it is not essential for triggering antiviral functions. After knocking out the STING or IRF3 gene, Mn²⁺ still retains its antiviral activity against HSV-1 and VSV. Furthermore, the results from transcriptomic analysis indicated that Mn²⁺ could induce a significantly change of apoptotic process in STING⁻/⁻ 3D4/21 cells. Mn²⁺ could induce cell apoptosis through oxidative stress pathway, and inhibiting the apoptotic signal could suppress the Mn²⁺-mediated antiviral activity in STING⁻/⁻ 3D4/21 cells. Additionally, dual knockout of IRF3 and caspase3 resulting in concurrent loss of IFN and apoptotic signals eliminate the antiviral effects of Mn²⁺. In summary, the current study has suggested that Mn²⁺ could exert antiviral effects not only through the cGAS-STING-IFN pathway but also via the ROS-apoptosis pathway.

Despite the importance of researcher literacy in veterinary postgraduate education, conventional training often overlooks the methodologies of evidence synthesis. This study assessed an evidence-based veterinary medicine (EBVM) pathway that integrates structured meta-analysis practice into the curriculum. Veterinary postgraduates were assigned to either an EBVM-intensive training group or a comparison group receiving routine instruction. Pre- and post-intervention assessments using a structured questionnaire revealed that the training group achieved superior proficiency in literature retrieval, critical appraisal, and methodological rigor. Notably, this pedagogical approach yielded nine peer-reviewed meta-analyses between 2021 and 2025, while no comparable output was observed in the comparison group. This evidence suggests that integrating hands-on meta-analysis into EBVM instruction serves as a catalyst, transforming theoretical learning into tangible, high-quality scholarly output.

Despite the importance of researcher literacy in veterinary postgraduate education, conventional training often overlooks the methodologies of evidence synthesis. This study assessed an evidence-based veterinary medicine (EBVM) pathway that integrates structured meta-analysis practice into the curriculum. Veterinary postgraduates were assigned to either an EBVM-intensive training group or a comparison group receiving routine instruction. Pre- and post-intervention assessments using a validated questionnaire revealed that the training group achieved superior proficiency in literature retrieval, critical appraisal, and methodological rigor. Notably, this pedagogical approach yielded nine peer-reviewed meta-analyses between 2021 and 2025, while no comparable output was observed in the comparison group. This evidence suggests that integrating hands-on meta-analysis into EBVM instruction serves as a catalyst, transforming theoretical learning into tangible, high-quality scholarly output.

The emergence of CRISPR/Cas9 technology has transformed the landscape of gene editing, allowing for precise alterations in DNA that hold great promise for research and potential therapies. However, a significant concern is the occurrence of off-target effects, which can lead to unintended genetic modifications with potentially harmful consequences. This paper explores the nature of off-target effects in CRISPR/Cas9, discussing how they arise and their implications for the reliability of gene editing. We identify the challenges faced in detecting and predicting these off-target interactions, including limitations in current detection techniques and the complexities of cellular biology. We present strategies aimed at minimizing off-target effects, such as careful design of guide RNAs, the use of computational tools for prediction, and improved delivery methods. Through a review of case studies, we highlight successful cases where off-target activity has been significantly reduced, offering insights into best practices for enhancing the accuracy of CRISPR/Cas9 applications. Moreover, we provide a comparative overview of Cas9, Cas12, and Cas13 systems, emphasizing their distinct target specificities, mechanisms of action, and off-target profiles. This comparison offers a broader understanding of how alternative CRISPR effectors may be leveraged to improve genome and transcriptome editing precision. This study underscores the importance of continued research to address the challenges of off-target effects, ultimately supporting the development of safer and more effective gene editing methods for clinical use.

African swine fever (ASF), induced by the African swine fever virus (ASFV), is an acute hemorrhagic disease characterized by high fever, systemic hemorrhages, and elevated mortality. Current diagnostic techniques including PCR and ELISA present limitations in field applications due to requirements for specialized equipment and prolonged processing duration. Therefore, rapid and accurate detection of ASFV has become a key link in ASF prevention and control. This study established a rapid and precise visual diagnostic approach by integrating the CRISPR/AapCas12b system with lateral flow strip (LFS) technology, specifically targeting the B646L gene encoding the major capsid protein p72. The CRISPR/AapCas12b-LFS platform achieved a sensitivity threshold of 6 copies/µL for B646L gene detection, completing analysis within an hour. Validation study confirmed exceptional specificity against common porcine pathogens including PRRSV, CSFV, PRV, PPV4, and PCV3. The developed assay demonstrated complete concordance with real-time PCR results when analyzing 34 clinical specimens including three heart samples, three liver samples, three spleen samples, three lung samples, three kidney samples, three lymph node samples, five serum samples, five blood samples, and five oral swab samples for ASFV detection. Overall, this method is sensitive, specific, and practicable onsite for ASFV detection, showing a great application potential for monitoring ASFV in the field.