Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Simultaneous management of intestinal mucosal barrier dysfunction and gut microbiota dysregulation represents a significant challenge in the treatment of inflammatory bowel disease (IBD). Herein, we report a novel system that integrates multi-enzyme mimicking cerium single-atom nanocatalysts (CeSACs) with Lactobacillus reuteri probiotics (LR@CeSACs) for multipronged management of IBD. In this system, CeSACs demonstrate robust multi-enzyme activities across a broad pH range, effectively scavenging elevated reactive oxygen species, downregulating pro-inflammatory cytokines, and suppressing the expression of fibrosis-related genes. Moreover, probiotics promote the targeting and retention of the CeSACs for sustained catalytic antioxidant therapy. In turn, the inflammation relief enabled by CeSACs promotes bacterial viability, allowing for the rapid reshaping of intestinal barrier function and the restoration of gut microbiota. Therefore, LR@CeSACs exhibit excellent catalytic anti-inflammatory and anti-fibrotic therapeutic effects, as well as a certain prophylactic effect, as demonstrated in several murine models.

Objective To investigate the dosimetric differences in preoperative short-course volumetric modulated arc therapy(VMAT)for rectal cancer using different fluence smoothing(FS)levels in the Monaco Treatment Planning System(Monaco TPS).Methods Twenty rectal cancer patients who received preoperative neoadjuvant short-course VMAT at some hospital from September 2021 to December 2022 were retrospectively selected.Four groups of radiotherapy plans were formulated using the Monaco TPS for each case,which were classified into an off group,a low group,a medium group and a high group based on the FS levels.Then the four groups were compared in terms of the dosimetric parameters,monitor unit and number of the segments in the planning target volume(PTV)and organ at risk(OAR).Statistical analysis was performed using SPSS 27.0 software.Results All the four groups had the doses to the target volume meeting clinical requirements,which had no significant differences in the doses to 5％(D5％)and 95％(D95％)to the target volume and the maximum dose(Dmax),minimum dose(Dmin),mean dose(Dmean)and conformity index(all P＞0.05).Statistical differences were found between the homogeneity indexes of the four groups(P＜0.05),with the medium group behaving the best.The number of the segments rose while the mornitor units decreased siginificantly with the increase of FS levels,with the differences being statistically significant(P＜0.05).There were no significant differences between the V25,V20,V15 and V10 of the small intestine,the V25 and V20 of the bladder and the V15 and V10 of the left and right femur(all P＞0.05).Conclusion In preoperative short-course VMAT for rectal cancer,clinical requirements can be met with different FS levels in the Monaco TPS,and medium-level FS results in optimal overall dose distribution in terms of treatment planning.[Chinese Medical Equipment Journal,2025,46(5):48-53]

Objective:To investigate lipid metabolism gene mutations and pathogenicity of hypertriglyceridemia acute pancreatitis (HTG-AP) patients.Methods:Clinical data of 495 HTG-AP patients admitted from June 2018 to June 2020 in the center for severe acute pancreatitis of Eastern Theater General Hospital were retrospectively analyzed. Whole-exome sequencing and mutation verification were performed by next-generation sequencing technology and Sanger sequencing. The pathogenicity of gene mutation was analyzed by population mutation ratio, pathogenicity prediction software, conservation scoring software, protein structure prediction, and in vitro experiments. Results:The mutation ratio of lipid metabolism-related genes, namely LPL, APOA5, LMF1, GPIHBP1, and APOC2, were 14.81%, 55.78%, 43.61%, 1.62%, and 0.61%, respectively. Among them, 44 heterozygous mutations in LPL gene were detected including 36 missense mutations, 5 nonsense mutations and 3 frameshift mutations, which were all rarely carried in single patient. Six HTG-AP patients carried the LPL gene heterozygous mutation c.835C>G (p.Leu279Val). The mean level of serum triglyceride at the onset of HTG-AP was 27.4 mmol/L. All of them had a history of recurrent HTG-AP, and most of them had severe acute pancreatitis. The serum LPL concentration and activity were lower than the normal level. The pathogenicity analysis results suggested that the LPL p.Leu279Val was a rare, highly possible pathogenic and highly conserved gene mutation. The in vitro results showed that the LPL p.Leu279Val could significantly reduce the synthesis and secretion ability of LPL as well as its enzymatic activity. Conclusions:The mutation ratio of lipid metabolism-related genes, including LPL, APOA5, LMF1, GPIHBP1, and APOC2, are relatively high in the HTG-AP patients. The LPL p.Leu279Val is a rare and highly possible pathogenic gene mutation, which may lead to recurrent episodes of HTG-AP.

Objective To investigate the dosimetric differences in preoperative short-course volumetric modulated arc therapy(VMAT)for rectal cancer using different fluence smoothing(FS)levels in the Monaco Treatment Planning System(Monaco TPS).Methods Twenty rectal cancer patients who received preoperative neoadjuvant short-course VMAT at some hospital from September 2021 to December 2022 were retrospectively selected.Four groups of radiotherapy plans were formulated using the Monaco TPS for each case,which were classified into an off group,a low group,a medium group and a high group based on the FS levels.Then the four groups were compared in terms of the dosimetric parameters,monitor unit and number of the segments in the planning target volume(PTV)and organ at risk(OAR).Statistical analysis was performed using SPSS 27.0 software.Results All the four groups had the doses to the target volume meeting clinical requirements,which had no significant differences in the doses to 5％(D5％)and 95％(D95％)to the target volume and the maximum dose(Dmax),minimum dose(Dmin),mean dose(Dmean)and conformity index(all P＞0.05).Statistical differences were found between the homogeneity indexes of the four groups(P＜0.05),with the medium group behaving the best.The number of the segments rose while the mornitor units decreased siginificantly with the increase of FS levels,with the differences being statistically significant(P＜0.05).There were no significant differences between the V25,V20,V15 and V10 of the small intestine,the V25 and V20 of the bladder and the V15 and V10 of the left and right femur(all P＞0.05).Conclusion In preoperative short-course VMAT for rectal cancer,clinical requirements can be met with different FS levels in the Monaco TPS,and medium-level FS results in optimal overall dose distribution in terms of treatment planning.[Chinese Medical Equipment Journal,2025,46(5):48-53]

Objective To observe the value of advanced patient motion correction(APMC)technology for improving imaging quality of low-dose pediatric chest CT.Methods Ninety-six children who received low-dose chest CT scan were retrospectively enrolled.CT images were reconstructed using full reconstruction technique(FULL)or APMC for both lung window and mediastinal window.Then the imaging qualities were subjectively evaluated according to image clarity,degree of motion artifacts,sharpness of structural edges and overall quality.The indexes of objective evaluation of imaging quality included the mean CT values of unit density pixel within ROI on lung window and mediastinal window,the standard deviation of CT values(SDCT)representing image noise,as well as signal-to-noise ratio(SNR).Sobel operator edge detection was performed on images,recording the mean gray level(G)and the standard deviation of gray value in Sobel(SDsobel)within ROI.Comparisons were made between FULL reconstruction and corrected images with APMC reconstruction in terms of subjective and objective evaluations,as well as parameters obtained from Sobel output images.Results Compared to FULL reconstruction,APMC corrected images showed reduced motion artifacts,improved edge structure sharpness and enhanced overall image quality(all P＜0.05),while there was no significant difference of image clarity(P＞0.05).Meanwhile,with APMC reconstructions,the mean CT values,SDCT on lung window and SDCT on mediastinal window increased(all P＜0.05),but there was no significant difference for the mean CT values on mediastinal window(P＞0.05),and SNR on both lung window and mediastinal window decreased(both P＜0.05).Besides,for Sobel output images,compared to FULL reconstruction,APMC reconstruction had increased G and SDsobel on both lung window and mediastinal window(all P＜0.05).Conclusion APMC technology applicated in low-dose chest CT scan of children could effectively reduce motion artifacts,improve edge clarity and imaging quality,hence enhance diagnostic sensitivity.

Objective To observe the value of advanced patient motion correction(APMC)technology for improving imaging quality of low-dose pediatric chest CT.Methods Ninety-six children who received low-dose chest CT scan were retrospectively enrolled.CT images were reconstructed using full reconstruction technique(FULL)or APMC for both lung window and mediastinal window.Then the imaging qualities were subjectively evaluated according to image clarity,degree of motion artifacts,sharpness of structural edges and overall quality.The indexes of objective evaluation of imaging quality included the mean CT values of unit density pixel within ROI on lung window and mediastinal window,the standard deviation of CT values(SDCT)representing image noise,as well as signal-to-noise ratio(SNR).Sobel operator edge detection was performed on images,recording the mean gray level(G)and the standard deviation of gray value in Sobel(SDsobel)within ROI.Comparisons were made between FULL reconstruction and corrected images with APMC reconstruction in terms of subjective and objective evaluations,as well as parameters obtained from Sobel output images.Results Compared to FULL reconstruction,APMC corrected images showed reduced motion artifacts,improved edge structure sharpness and enhanced overall image quality(all P＜0.05),while there was no significant difference of image clarity(P＞0.05).Meanwhile,with APMC reconstructions,the mean CT values,SDCT on lung window and SDCT on mediastinal window increased(all P＜0.05),but there was no significant difference for the mean CT values on mediastinal window(P＞0.05),and SNR on both lung window and mediastinal window decreased(both P＜0.05).Besides,for Sobel output images,compared to FULL reconstruction,APMC reconstruction had increased G and SDsobel on both lung window and mediastinal window(all P＜0.05).Conclusion APMC technology applicated in low-dose chest CT scan of children could effectively reduce motion artifacts,improve edge clarity and imaging quality,hence enhance diagnostic sensitivity.

Objective:To investigate lipid metabolism gene mutations and pathogenicity of hypertriglyceridemia acute pancreatitis (HTG-AP) patients.Methods:Clinical data of 495 HTG-AP patients admitted from June 2018 to June 2020 in the center for severe acute pancreatitis of Eastern Theater General Hospital were retrospectively analyzed. Whole-exome sequencing and mutation verification were performed by next-generation sequencing technology and Sanger sequencing. The pathogenicity of gene mutation was analyzed by population mutation ratio, pathogenicity prediction software, conservation scoring software, protein structure prediction, and in vitro experiments. Results:The mutation ratio of lipid metabolism-related genes, namely LPL, APOA5, LMF1, GPIHBP1, and APOC2, were 14.81%, 55.78%, 43.61%, 1.62%, and 0.61%, respectively. Among them, 44 heterozygous mutations in LPL gene were detected including 36 missense mutations, 5 nonsense mutations and 3 frameshift mutations, which were all rarely carried in single patient. Six HTG-AP patients carried the LPL gene heterozygous mutation c.835C>G (p.Leu279Val). The mean level of serum triglyceride at the onset of HTG-AP was 27.4 mmol/L. All of them had a history of recurrent HTG-AP, and most of them had severe acute pancreatitis. The serum LPL concentration and activity were lower than the normal level. The pathogenicity analysis results suggested that the LPL p.Leu279Val was a rare, highly possible pathogenic and highly conserved gene mutation. The in vitro results showed that the LPL p.Leu279Val could significantly reduce the synthesis and secretion ability of LPL as well as its enzymatic activity. Conclusions:The mutation ratio of lipid metabolism-related genes, including LPL, APOA5, LMF1, GPIHBP1, and APOC2, are relatively high in the HTG-AP patients. The LPL p.Leu279Val is a rare and highly possible pathogenic gene mutation, which may lead to recurrent episodes of HTG-AP.

Objective:To investigate the effect of Eriocitrin on the proliferation of esophageal cancer KYSE30 cells,and to explore its possible mechanism based on ferroptosis.Methods:Esophageal cancer KYSE30 cells were divided into 8 groups:the control group(conventional culture),RSL3 group(treated with 3 μmol/L ferroptosis inducer RSL3),Eriocitrin group(treated with 75 μmol/L Eriocitrin),Eriocitrin+Fer-1 group(treated with 5 μmol/L of ferroptosis inhibitor Fer-1 and 75 μmol/L Eriocitrin),Fer-1 group(treated with 5 μmol/L Fer-1 treatment),oe-NC group(transfected with blank vector control),oe-STAT3 group(transfected with STAT3 overexpression vector)and oe-STAT3+Eriocitrin group(transfected with STAT3 overexpression vector and then treated with 75 μmol/L Eriocitrin).Proliferation abilities of cells in each group were detected using CCK-8 assay,EdU incorporation assay and clone formation assay respectively.The levels of intracellular ferroptosis-related indicators were detected using the ELISA kits.Western blotting was used to detect the expression of STAT3/GPX4 pathway-related proteins.KYSE30 cell nude mouse subcutaneous transplanted tumor model was constructed to observe the effects of Eriocitrin and Fer-1 on the growth of transplanted tumors.Results:Eriodictyol could inhibit the proliferation and clone formation of KYSE30 cells,increase the levels of reactive oxygen species(ROS),malondialdehyde(MDA)and Fe2+,decrease the level of glutathione(GSH)(all P<0.05)and suppress the growth of transplanted tumors in nude mice.These effects could be reversed by Fer-1(P<0.05).Overexpression of STAT3 could abolish the inductive effect of eriodictyol on ferroptosis and its inhibitory effect on the STAT3/GPX4 pathway(P<0.05).Conclusion:Eriocitrin could induce ferroptosis in esophageal cancer KYSE30 cells by inhibiting STAT3/GPX4 signaling pathway and exert significant antitumor effects in esophageal cancer.

The aim of this study was to compare the immune responses of C57BL/6 mice immunized with two pathogens, foot-and-mouth disease virus (FMDV) and Senecavirus A (SVA), and to provide clues for revealing the regulatory mechanisms of acquired immunity. Inactivated and purified FMDV and SVA antigens were used to immunize C57BL/6 mice respectively, and the mice immunized with PBS were taken as the control. The percentages of Th1 and Th2 cells in the spleen lymphocytes of mice in each group were analyzed by flow cytometry at 14 and 28 days after immunization. RNA-Seq was performed for the spleen. Mouse macrophages were stimulated with the antigens in vitro to examine the expression of the differentially expressed genes (DEGs) screened out. The results showed that 14 days after immunization, there was no significant difference in the magnitude of the Th1/Th2 immune response elicited by the FMDV and SVA antigens. After 28 days, the magnitudes of the Th1 and Th2 immune responses elicited by the SVA antigen were higher than those elicited by the FMDV antigen. RNA-Seq revealed two common DEGs, Rsad2 and Tspan8, between the two immunization groups, which indicated that the two genes may be involved in the activation of the Th1/Th2 immune responses by FMDV and SVA antigens. FMDV and SVA antigens stimulated macrophages to secrete interleukin (IL)-12 and IL-33 in vitro, and the expression of Tspan8 and Rsad2 was consistent with the RNA-Seq results. The expression of Rsad2 was regulated by type I interferons (IFNα, IFNβ). In this study, we obtained the DEGs involved in the immune responses to the two antigens in mouse spleen, which provides a molecular basis for investigating the immune response mechanisms induced by FMDV and SVA.
Animals ; Foot-and-Mouth Disease Virus/genetics* ; Mice ; Spleen/cytology* ; Mice, Inbred C57BL ; Antigens, Viral/genetics* ; Transcriptome ; Th1 Cells/immunology* ; Immunization ; Viral Vaccines/immunology* ; Th2 Cells/immunology* ; Foot-and-Mouth Disease/immunology* ; Interleukin-33/genetics* ; Female ; Macrophages/immunology* ; Picornaviridae