Dipeptidyl peptidase 4(DPP4)is one of the binding receptors for the spike(S)protein of porcine hemagglutinating encephalomyelitis virus(PHEV).To identify the key amino acid binding sites at the interface of DPP4 protein and PHEV spike protein and explore the impact of their mu-tations on viral infection,recombinant plasmids of porcine DPP4,murine DPP4 and human DPP4(pDPP4,mDPP4,hDPP4)and spike protein truncations(S311-608,S13-298)were constructed and co-transfected into HEK293T cells to detect the protein binding by co-immunoprecipitation(CoIP).Simultaneously,the expression of PHEV proteins and genes was detected in DPP4-over-expressing HeLa cells infected by PHEV using Western blot and RT-qPCR.homologues were overexpressed in HeLa cells which were not susceptible to and then inoculated with the virus,and.Subsequently,the important pDPP4 amino acid sites at the interaction interface were mutated one by one using a point mutation kit to construct mutant overexpression plasmids.The mutant and wild-type pDPP4 were co-transfected into HEK293T cells with the spike protein truncations re-spectively to assay the protein interaction ability by co-immunoprecipitation.After HeLa cells over-expressing the mutant and wild-type pDPP4 were infected by PHEV,the replication level of PHEV was detected by Western blot and RT-qPCR.Compared with hDPP4,the pDPP4 and mDPP4 had significantly stronger binding to PHEV spike protein,which significantly promote PHEV infection.Moreover,mutation of pDPP4 glycosylation sites significantly enhanced the inter-action with PHEV spike protein,and the mutation of glycosylation sites(N229,N321)dramatical-ly promoted PHEV infection.The above results indicated that DPP4 glycosylation modification plays an important shielding role in the process of PHEV invading target cells mediated by spike protein.

Objective To assess the efficacy of combining specialized scenarios with workshop-based teaching in training newly recruited nurses for occupational exposure.Methods This study was conducted with 200 nurses recruited between January 2020 and December 2021 as the control group and 200 nurses from January 2022 to December 2023 as the intervention group.The control group and the intervention group received routine training and workshop-based training,respectively.The efficacy was assessed through theoretical exams,skills assessments,occupational exposure incidence,and training satisfaction,measured u-sing the Simulation Design Scale(SDS).Results The intervention group demonstrated higher scores on theoretical knowledge and operational skills assessments compared to the control group.The occupational exposure were lower in the intervention group.Additionally,the intervention group scored higher on the SDS and reported greater satisfaction with the training(P＜0.05).Conclusion The teaching method integrated with specialized scenarios and workshop-based teaching effectively enhances the theoretical knowledge and operational skills related to occupational exposure among newly recruited nurses.It reduces the inci-dence of occupational exposure,such as needlestick injuries,and improves training outcomes and nurse satisfaction,making it a valuable approach for clinical application.

Dipeptidyl peptidase 4(DPP4)is one of the binding receptors for the spike(S)protein of porcine hemagglutinating encephalomyelitis virus(PHEV).To identify the key amino acid binding sites at the interface of DPP4 protein and PHEV spike protein and explore the impact of their mu-tations on viral infection,recombinant plasmids of porcine DPP4,murine DPP4 and human DPP4(pDPP4,mDPP4,hDPP4)and spike protein truncations(S311-608,S13-298)were constructed and co-transfected into HEK293T cells to detect the protein binding by co-immunoprecipitation(CoIP).Simultaneously,the expression of PHEV proteins and genes was detected in DPP4-over-expressing HeLa cells infected by PHEV using Western blot and RT-qPCR.homologues were overexpressed in HeLa cells which were not susceptible to and then inoculated with the virus,and.Subsequently,the important pDPP4 amino acid sites at the interaction interface were mutated one by one using a point mutation kit to construct mutant overexpression plasmids.The mutant and wild-type pDPP4 were co-transfected into HEK293T cells with the spike protein truncations re-spectively to assay the protein interaction ability by co-immunoprecipitation.After HeLa cells over-expressing the mutant and wild-type pDPP4 were infected by PHEV,the replication level of PHEV was detected by Western blot and RT-qPCR.Compared with hDPP4,the pDPP4 and mDPP4 had significantly stronger binding to PHEV spike protein,which significantly promote PHEV infection.Moreover,mutation of pDPP4 glycosylation sites significantly enhanced the inter-action with PHEV spike protein,and the mutation of glycosylation sites(N229,N321)dramatical-ly promoted PHEV infection.The above results indicated that DPP4 glycosylation modification plays an important shielding role in the process of PHEV invading target cells mediated by spike protein.

Objective To assess the efficacy of combining specialized scenarios with workshop-based teaching in training newly recruited nurses for occupational exposure.Methods This study was conducted with 200 nurses recruited between January 2020 and December 2021 as the control group and 200 nurses from January 2022 to December 2023 as the intervention group.The control group and the intervention group received routine training and workshop-based training,respectively.The efficacy was assessed through theoretical exams,skills assessments,occupational exposure incidence,and training satisfaction,measured u-sing the Simulation Design Scale(SDS).Results The intervention group demonstrated higher scores on theoretical knowledge and operational skills assessments compared to the control group.The occupational exposure were lower in the intervention group.Additionally,the intervention group scored higher on the SDS and reported greater satisfaction with the training(P＜0.05).Conclusion The teaching method integrated with specialized scenarios and workshop-based teaching effectively enhances the theoretical knowledge and operational skills related to occupational exposure among newly recruited nurses.It reduces the inci-dence of occupational exposure,such as needlestick injuries,and improves training outcomes and nurse satisfaction,making it a valuable approach for clinical application.

Background/Aims: Steatotic liver disease (SLD) is a common manifestation in chronic hepatitis C (CHC). Metabolic alterations in CHC are associated with metabolic dysfunction-associated steatotic liver disease (MASLD). We aimed to elucidate whether hepatitis C virus (HCV) eradication mitigates MASLD occurrence or resolution. Methods: We enrolled 5,840 CHC patients whose HCV was eradicated by direct-acting antivirals in a nationwide HCV registry. MASLD and the associated cardiometabolic risk factors (CMRFs) were evaluated at baseline and 6 months after HCV cure. Results: There were 2,147 (36.8%) patients with SLD, and 1,986 (34.0%) of them met the MASLD criteria before treatment. After treatment, HbA1c (6.0% vs. 5.9%, p<0.001) and BMI (24.8 kg/m2 vs. 24.7 kg/m2, p<0.001) decreased, whereas HDL-C (49.1 mg/dL vs. 51.9 mg/dL, p<0.001) and triglycerides (102.8 mg/dL vs. 111.9 mg/dL, p<0.001) increased significantly. The proportion of patients with SLD was 37.5% after HCV eradication, which did not change significantly compared with the pretreatment status. The percentage of the patients who had post-treatment MASLD was 34.8%, which did not differ significantly from the pretreatment status (p=0.17). Body mass index (BMI) (odds ratio [OR] 0.89; 95% confidence intervals [CI] 0.85–0.92; p<0.001) was the only factor associated with MASLD resolution. In contrast, unfavorable CMRFs, including BMI (OR 1.10; 95% CI 1.06–1.14; p<0.001) and HbA1c (OR 1.19; 95% CI 1.04–1.35; p=0.01), were independently associated with MASLD development after HCV cure. Conclusions HCV eradication mitigates MASLD in CHC patients. CMRF surveillance is mandatory for CHC patients with metabolic alterations, which are altered after HCV eradication and predict the evolution of MASLD.

The development of drug-resistant influenza and new pathogenic virus strains underscores the need for antiviral therapeutics. Currently, neuraminidase (NA) inhibitors are commonly used antiviral drugs approved by the US Food and Drug Administration (FDA) for the prevention and treatment of influenza. Here, we show that vitisin B (VB) inhibits NA activity and suppresses H1N1 viral replication in MDCK and A549 cells. Reactive oxygen species (ROS), which frequently occur during viral infection, increase virus replication by activating the NF-κB signaling pathway, downmodulating glucose-6-phosphate dehydrogenase (G6PD) expression, and decreasing the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant response activity. VB decreased virus-induced ROS generation by increasing G6PD expression and Nrf2 activity, and inhibiting NF-κB translocation to the nucleus through IKK dephosphorylation. In addition, VB reduced body weight loss, increased survival, decreased viral replication and the inflammatory response in the lungs of influenza A virus (IAV)-infected mice. Taken together, our results indicate that VB is a promising therapeutic candidate against IAV infection, complements existing drug limitations targeting viral NA. It modulated the intracellular ROS by G6PD, Nrf2 antioxidant response pathway, and NF-κB signaling pathway. These results demonstrate the feasibility of a multi-targeting drug strategy, providing new approaches for drug discovery against IAV infection.

Background/Aims: Little is known about the clinical course of hepatitis B virus (HBV)-infected patients undergoing anti-tumor necrosis factor α (TNF-α) therapy for inflammatory bowel disease (IBD). We aimed to investigate the clinical course of HBV infection and IBD and to analyze liver dysfunction risks in patients undergoing anti-TNF-α therapy. Methods: This retrospective multinational study involved multiple centers in Korea, China, Tai-wan, and Japan. We enrolled IBD patients with chronic or resolved HBV infection, who received anti-TNF-α therapy. The patients’ medical records were reviewed, and data were collected using a web-based case report form. Results: Overall, 191 patients (77 ulcerative colitis and 114 Crohn’s disease) were included, 28.3% of whom received prophylactic antivirals. During a median follow-up duration of 32.4 months, 7.3% of patients experienced liver dysfunction due to HBV reactivation. Among patients with chronic HBV infection, the proportion experiencing liver dysfunction was significantly higher in the non-prophylaxis group (26% vs 8%, p=0.02). Liver dysfunction occurred in one patient with resolved HBV infection. Antiviral prophylaxis was independently associated with an 84% reduction in liver dysfunction risk in patients with chronic HBV infection (odds ratio, 0.16; 95% confidence interval, 0.04 to 0.66; p=0.01). The clinical course of IBD was not associated with liver dysfunction or the administration of antiviral prophylaxis. Conclusions Liver dysfunction due to HBV reactivation can occur in HBV-infected IBD patients treated with anti-TNF-α agents. Careful monitoring is needed in these patients, and antivirals should be administered, especially to those with chronic HBV infection.

Duloxetine and thioctic acid (TA) are standard drugs for treating diabetic neuropathy, a primary complication associated with diabetes. In this study, ultra performance liquid chromatography coupled with tandem mass spectrometry methods was successfully developed and validated for quantifying duloxetine and TA in biological samples. The protein precipitation method was used to extract duloxetine, TA and their internal standards from beagle dog plasma. A Hypersil Gold C18 column (150 × 2.1 mm, 1.9 μm) was used for the experiment. Isocratic elution with 0.1% formic acid in acetonitrile (A) and 0.1% formic acid (B) was used for duloxetine, whereas a gradient elution with 0.03% acetic acid (A) and acetonitrile (B) was used for TA. The validated parameters included linearity, sensitivity, accuracy, precision, selectivity, matrix effect, stability, and recovery under different conditions. The linear ranges of the calibration curves for duloxetine and TA were 5–800 ng/mL and 5–1,000 ng/mL, respectively. An intra- and inter-run precision of ± 15% can be observed in all quality control samples. These methods were successfully used for pharmacokinetics (PKs) studies in beagle dogs to compare PK differences in a fixed-dose combination including duloxetine and TA and co-administration of the 2 drugs.

Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.
Agaricales ; Gastrodia/genetics* ; Protoplasts ; Symbiosis/genetics* ; Transformation, Genetic

Aim To explore the regulatory effect of Danggui-chuanxiong herb pair (GX) on JAK-STAT signaling pathway in rats with cerebral ischemia/reper-fusion injury (I/R). Methods The I/R injury rat model was constructed by modified suture occlusion method. After 24 hours of perfusion, Zea Longa scoring method was used to score the neurological function, TTC staining to detect the cerebral infarct volume of rats, HE staining to observe the pathological changes of brain tissues, the biochemical method to determine the MDA, SOD, GSH-Px expression, ELISA to detect the expression of NF-κB, VEGF, ICAM-1 and PAH in brain tissues, and immunohistochemical method to detect JAK2, p-STAT3, AKT And ERK1/2 expression of the brain tissue ischemic penumbra area. Results Compared with sham group, model rats had severe neurological damage, larger cerebral infarction, necrosis, edema, inflammation, disorder of nerve cell arrangement, abnormal cell enlargement, vacuole-like changes, neuron reduction and other pathologies in brain tissues. The expression JeveJs of MDA, NF-κB, VEGF, PAI-1 and ICAM-1 in brain tissues of model group significantly increased, and the expression levels of GSH-Px and SOD were significantly reduced. Compared with model group, the neurological scores of rats in GX