Porcine Circovirus 2 (PCV2) is a crucial swine pathogen and considered a primary causative agent of porcine circovirus-associated diseases (PCVADs), posing a serious economic threat to the swine industry across globe. The world’s biggest agricultural conglomerates have teamed up to create giant commercial pig farms across Shanghai due to the proximity of this region to more affluent lean-pork markets. Since its discovery, PCV2 has displayed extraordinary genetic diversity, and its genome is swiftly evolving through a series of mutations and recombinations. However, limited information on epidemiology, molecular characteristics, vaccine cross-protection, and the co-infection rate of PCV2 with other lethal swine diseases can adversely impact the pig production in the region. To investigate the molecular epidemic characteristics and genetic evolution of PCV2, pigs with doubtful symptoms of PCVADs were sampled from various commercial pig farms with a history of PWMS and/or PDNS across Shanghai from 2014 to 2018. Our results revealed the coexistence of multiple PCV2 genotypes (PCV2b, PCV2e, and PCV2d) among Shanghai pig herds and dominance of PCV2d among them. We also found critical amino acid substitutions in epitope regions of important capsid proteins in PCV2 isolates involved in viral replication and host immune escape. Spotted mutations may favor the prevalence and survival of various PCV2 genotypes despite availability of commercial vaccines. This study also provides insight into the co-infection status of PCV2 with major lethal swine viral diseases such as PPV and PPRSV. Collectively, these investigations will contribute to understanding the molecular epidemiology and evolution of PCV2 across the region.

Purpose Avascular necrosis (AVN) of the femoral head is a common orthopaedic disease that is difficult to treat. The purpose of this study was to explore the preliminary efficacy of a self-designed umbrella-shaped memory alloy femoral head support device in the treatment of adult patients with avascular osteonecrosis of the femoral head. Methods The minimally-invasive approach involved curettage of the necrotic tissue of the femoral head, and a self-designed umbrella-shaped, memory alloy femoral head support device was implanted into the collapsed necrotic area to support the collapsed femoral head. Autologous iliac bone and artificial bone were implanted into the support device for the treatment of adult patients with avascular osteonecrosis of the femoral head. Results The clinical device was used in ten patients and 18 hip joints. The support device failed in one hip joint, which subsequently underwent joint replacement surgery, and the remaining 17 implanted devices were followed up for four to 19 months. The 17 postoperative hip joints were evaluated using the percent-efficacy evaluation method for avascular osteonecrosis of the femoral head in adult patients, and the efficacy rate was 82.35 %. Conclusion The umbrella-shaped femoral head support device can be used in Ficat stage I, stage II, and stage III adult patients with avascular osteonecrosis of the femoral head.

Nonstructural protein 2A (NS2A) of the Japanese encephalitis virus (JEV) contributes to viral replication and pathogenesis; however, a lack of NS2A-specific antibodies restricts studies on the underlying mechanisms. In this study, we constructed a recombinant JEV with a hemagglutinin (HA)-tagged NS2A (JEV-HA/NS2A/∆NS1’) to overcome this challenge. An HA-tag was fused to the N-terminus of NS2A (HA-NS2A) at the intergenic junction between NS1 and NS2A. A peptide linker, “FNG”, was added to the N-terminus of HA-tag to ensure correct cleavage between the C-terminus of NS1 and the N-terminus of HA-NS2A. To avoid the side effects of an unwanted NS1’ tagged with HA (HA-NS1’), an alanine-to-proline (A30P) substitution was introduced at residue 30 of NS2A to abolish HA-NS1’ production. The HA-tag insertion and A30P substitution were stably present in JEV-HA/NS2A/∆NS1’ after six passages and did not exhibit any significant effects on viral replication and plaque morphology. Taking advantage of HA-NS2A, we examined the activities of NS2A during JEV infection in vitro using anti-HA antibodies. NS2A was observed to be localized to the endoplasmic reticulum and interact with viral NS2B and NS3 during virus infection. These data suggest that JEV-HA/NS2A/∆NS1’ can serve as a model for the analysis of the biological characteristics and functions of NS2A in vitro during JEV infection.

Caprine arthritis encephalitis is an infectious disease caused by the caprine arthritis encephalitis virus that infects goats, sheep, and other small ruminants. An outbreak of CAEV could be extremely harmful to the goat farming industry and could cause severe economic losses. We designed specific primers and probes for the gag gene and established a TaqMan real-time quantitative polymerase chain reaction assay. This method’s correlation coefficient (R²) was >0.999, and the sensitivity of the assay to the plasmid-carried partial gag gene was approximately 10 copies/µL, 1000 times higher than that of conventional PCR. No specific fluorescence was detected for other sheep viruses. Using this method, we tested 776 asymptomatic sheep blood samples and 4 neurodegenerative sheep brain samples from six farms in eastern China, and the positivity rate was 0.77% (6/780). The gag gene was partially sequenced in the three positive samples and compared with the sequences from other representative strains in GenBank. The results revealed that all three strains belonged to the B1 subtype and were most closely related to the strains from Shanxi and Gansu, previously isolated in China, with their homology ranging from 97.7% to 98.9%. These results suggest that the designed RT-qPCR assay can be used to detect subclinical CAEV in sheep and that the virus is still present in eastern China.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses in swine production worldwide, characterized by reproductive failure and respiratory disease. In China, the cocirculation of diverse strains drives ongoing viral evolution and the emergence of novel recombinants, posing serious challenges for disease control. Here, we report the isolation and characterization of a novel PRRSV strain (JS‐YZ/2023) from a PRRS‐suspected pig farm. Genomic analysis revealed a unique recombinant pattern between NADC30‐like and HP‐PRRSV lineages, distinct from previously reported recombinants. Experimental infection in piglets demonstrated significant pathogenicity, with persistent fever, severe anorexia, progressive weight loss, and pathological lesions including interstitial pneumonia with lymphocyte infiltration and marked disruption of intestinal villi. Notably, JS‐YZ/2023 exhibited strong intestinal tropism, inducing a local cytokine storm in the small intestine and showing PRRSV‐N protein antigen accumulation in crypt epithelial cells, as detected by immunohistochemistry. In addition, we successfully constructed a full‐length infectious cDNA clone of JS‐YZ/2023 incorporating a green fluorescent protein (GFP) reporter gene (rGFP‐JS‐YZ/2023). This engineered virus provides a valuable tool for investigating the pathogenic mechanisms of NADC30‐like recombinants and facilitating vaccine development.

Viral replicons are efficient tools to understand the mechanisms of viral replication and screen antiviral drugs. In this study, a viral-cDNA-based replicon of Japanese encephalitis virus (JEV), which is the causative agent of Japanese encephalitis, was constructed by replacing the viral structural proteins with a green fluorescent protein (JEV-GFP replicon). The resulting JEV-GFP replicon was used as a tool to screen antiviral drugs targeting JEV nonstructural proteins, and the five compounds JNJ-A07, HZ-1157, NITD-2, quinine, and NITD008 were obtained, which significantly inhibited the replication of the JEV-GFP replicon and JEV in vitro, and the properties of these five compounds were also analyzed. The CC₅₀, EC₅₀, and SI indices of these five compounds were analyzed. In addition, the JEV-GFP replicon was used as a tool to identify the residues of viral nonstructural proteins involved in RNA replication, and the cysteine residue at position 4 of nonstructural protein 1 was found to be essential for JEV RNA replication. These data suggested that the noninfectious JEV-GFP replicon could be used as tool for different purposes, such as antiviral drug screening and gene function studies.