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*

Objective:To investigate the prevalence of sarcopenia and potential influencing factors in middle-aged and elderly populations in Zhejiang Province.Methods:Data were obtained from Zhejiang Provincial Household Economic Status Survey, a cross-sectional survey was condcuted in middle-aged and olde adults selected through multi-stage sampling in three cities in Zhejiang (Huzhou, Jiaxing and Shaoxing) in July 2023. A total of 3 019 study participants, average age 62.3 years old, 53.5% men, were included according to the inclusion and exclusion criteria. Sarcopenia screening was conducted by using the questionnaire with five sarcopenia related-items. Univariable and multivariable logistic regression analysis was used to identify the factors associated with sarcopenia.Results:The prevalence of sarcopenia in the middle-aged and old study participants was 4.47%. Significant differences were observed between the participants with or without sarcopenia in terms of age, educational level, BMI, alcohol consumption status, diet habit, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, adulthood community socioeconomic status, muscle strength, walking assistance, ability to stand from seat, ability to climb stairs, and fall frequency ( P<0.05). Multivariable logistic regression analysis revealed that old age (≥75 years: OR=2.82, 95% CI: 1.60-4.97), low body weight ( OR=1.96, 95% CI: 1.06-3.62), unhealthy diet habit ( OR=1.57, 95% CI: 1.01-2.46), physical inactivity ( OR=5.80, 95% CI: 3.09-10.88), poor or very poor sleep quality ( OR=1.65, 95% CI:1.23-2.41), number of chronic diseases (1 chronic disease: OR=1.84, 95% CI: 1.08-3.14; 2 chronic diseases: OR=3.22, 95% CI: 1.81-5.71; 3 or more chronic diseases: OR=3.74, 95% CI: 2.11-6.65), poor childhood socioeconomic status ( OR=2.98, 95% CI: 1.23-7.20), and poor adulthood community socioeconomic status ( OR=3.87, 95% CI: 1.63-9.17) were significant risk factors for sarcopenia. Conclusions:The prevalence of sarcopenia was relatively low in middle-aged and old population in Zhejiang. Age, BMI, unhealthy diet, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, and adulthood community socioeconomic status were identified as significant influencing factors.

To explore the effects of different exercise prescriptions on glycemic metabolism in East Asian patients with type 2 diabetes mellitus (T2DM) and to compare the differences in the impact of population characteristics and exercise components on glycemic metabolism.

Objective:To explore the correlations of sleep quality and architecture with heart rate variability (HRV) in patients with stenoses of vertebrobasilar artery system and internal carotid artery system.Methods:A retrospective study was performed; 72 patients with stenosis or occlusion of the head and neck arteries (not resulting in cerebral infarction) admitted to Department of Neurology, Second Affiliated Hospital of Zhengzhou University from June 2023 to June 2024 were chosen, including 33 patients with moderate-to-severe stenosis or occlusion of the vertebrobasilar system (VB group) and 39 patients with moderate-to-severe stenosis or occlusion of the internal carotid artery system (ICA group). Pittsburgh sleep quality index (PSQI) and polysomnography (PSG) were used to evaluate the sleep quality and architecture, respectively; and 24-hour ambulatory electrocardiogram was used to assess the HRV. Differences in PSQI score, PSG and HRV parameters between the two groups were compared; partial correlation analysis was used to explore the correlations of HRV parameters with PSQI scores and PSG parameters; multivariate linear regression was used to analyze the independent influencing factors for HRV.Results:(1) Compared with the ICA group, the VB group exhibited significantly higher PSQI scores, spontaneous arousal index (SAI), ratio of time of stage 1 non-rapid eye movement sleep/total sleep time (T N1/T t), and apnea-hypopnea index (AHI), while significantly lower ratio of time of rapid eye movement sleep/total sleep time (T R/T t), spindle wave density in stage 2 non-rapid eye movement sleep (N2), lowest blood oxygen saturation, standard deviation of normal to normal intervals (SDNN) of all sinus beats, low-frequency power (LF), and high-frequency power (HF, P<0.05). (2) In both VB group and ICA group, SDNN was negatively correlated with PSQI score ( r=-0.461, P=0.020; r=-0.378, P=0.036). In the VB group, SDNN was negatively correlated with T N1/T t ( r=-0.467, P=0.019) and SAI ( r=-0.551, P=0.004), and positively correlated with ratio of time of stage 3 non-rapid eye movement sleep/total sleep time (T N3/T t, r=0.686, P<0.001) and spindle wave density in N2 ( r=0.518, P=0.008); LF and HF were negatively correlated with SAI ( r=-0.481, P=0.015; r=-0.564, P=0.003). In the ICA group, HF was negatively correlated with spindle wave density in N2 ( r=-0.369; P=0.041). (3) Multivariate linear regression results indicated that T N3/T t (β=0.348, P=0.018), SAI (β=-0.330, P=0.018), and spindle wave density in N2 (β=0.286, P=0.013) were independent influencing factors for Ln_SDNN in patients with moderate-to-severe stenosis or occlusion of the vertebrobasilar system. Conclusion:Patients with stenosis or occlusion of the vertebrobasilar system exhibit poorer subjective sleep quality, increased light sleep, heightened arousal, and reduced sleep stability compared with those with stenosis or occlusion of the internal carotid artery system, which may be caused by the imbalance of autonomic nerve function.

Objective To investigate the effects of Aurora kinase A(AURKA)on the tumor microenvironment of colorectal cancer(CRC)and to predict the active compounds in Chinese herbs that can target AURKA.Methods Based on the transcriptomic data and clinical information from 380 CRC tissues and 51 paracancerous tissues in The Cancer Genome Atlas(TCGA)database,the infiltration of different cells in the tumor tissues was analyzed using xCell and the binding of active compounds of Chinese herbs with AURKA was predicted through molecular docking.Results The expression of AURKA was significantly upregulated in CRC tissues compared with that in paracancerous tissues(P＜0.05),and CRC patients with high AURKA expression had shorter overall survival.Compared with the AURKA low-expression group,the abundance of macrophages,monocytes,and effector memory CD4+and CD8+T cells was significantly downregulated in the AURKA high-expression group(P＜0.05).In addition,the cytotoxicity of T cells was significantly reduced(P＜0.05).Further analysis revealed that AURKA expression was positively correlated with the abundance of myeloid-derived suppressor cells(MDSCs)and the expression levels of their chemokines CXCL2 and CXCL5(P＜0.05).Genes that were differentially expressed between the AURKA high-and low-expression groups were mainly enriched in monocyte migration,chemokine-induced cellular responses,and other related processes.Chinese herbal compounds,including hesperidin,aristololactam A Ⅱ a,anacardic acid,coumestrol,and 17β-estradiol,all showed binding energies to AURKA lower than-1.2 kcal/mol,indicating a certain level of binding stability.Among these Chinese herbal compounds,17β-estradiol exhibited the best binding stability to AURKA-3UOL.Conclusion The high expression of AURKA in CRC tissues suggests a poor clinical prognosis.AURKA can promote the development of a suppressive immune microenvironment in CRC,and 17β-estradiol,an active compound from Chinese herbs,is a potential therapeutic agent targeting AURKA.

Objective:To investigate the prevalence of sarcopenia and potential influencing factors in middle-aged and elderly populations in Zhejiang Province.Methods:Data were obtained from Zhejiang Provincial Household Economic Status Survey, a cross-sectional survey was condcuted in middle-aged and olde adults selected through multi-stage sampling in three cities in Zhejiang (Huzhou, Jiaxing and Shaoxing) in July 2023. A total of 3 019 study participants, average age 62.3 years old, 53.5% men, were included according to the inclusion and exclusion criteria. Sarcopenia screening was conducted by using the questionnaire with five sarcopenia related-items. Univariable and multivariable logistic regression analysis was used to identify the factors associated with sarcopenia.Results:The prevalence of sarcopenia in the middle-aged and old study participants was 4.47%. Significant differences were observed between the participants with or without sarcopenia in terms of age, educational level, BMI, alcohol consumption status, diet habit, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, adulthood community socioeconomic status, muscle strength, walking assistance, ability to stand from seat, ability to climb stairs, and fall frequency ( P<0.05). Multivariable logistic regression analysis revealed that old age (≥75 years: OR=2.82, 95% CI: 1.60-4.97), low body weight ( OR=1.96, 95% CI: 1.06-3.62), unhealthy diet habit ( OR=1.57, 95% CI: 1.01-2.46), physical inactivity ( OR=5.80, 95% CI: 3.09-10.88), poor or very poor sleep quality ( OR=1.65, 95% CI:1.23-2.41), number of chronic diseases (1 chronic disease: OR=1.84, 95% CI: 1.08-3.14; 2 chronic diseases: OR=3.22, 95% CI: 1.81-5.71; 3 or more chronic diseases: OR=3.74, 95% CI: 2.11-6.65), poor childhood socioeconomic status ( OR=2.98, 95% CI: 1.23-7.20), and poor adulthood community socioeconomic status ( OR=3.87, 95% CI: 1.63-9.17) were significant risk factors for sarcopenia. Conclusions:The prevalence of sarcopenia was relatively low in middle-aged and old population in Zhejiang. Age, BMI, unhealthy diet, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, and adulthood community socioeconomic status were identified as significant influencing factors.

Objective:To explore the correlations of sleep quality and architecture with heart rate variability (HRV) in patients with stenoses of vertebrobasilar artery system and internal carotid artery system.Methods:A retrospective study was performed; 72 patients with stenosis or occlusion of the head and neck arteries (not resulting in cerebral infarction) admitted to Department of Neurology, Second Affiliated Hospital of Zhengzhou University from June 2023 to June 2024 were chosen, including 33 patients with moderate-to-severe stenosis or occlusion of the vertebrobasilar system (VB group) and 39 patients with moderate-to-severe stenosis or occlusion of the internal carotid artery system (ICA group). Pittsburgh sleep quality index (PSQI) and polysomnography (PSG) were used to evaluate the sleep quality and architecture, respectively; and 24-hour ambulatory electrocardiogram was used to assess the HRV. Differences in PSQI score, PSG and HRV parameters between the two groups were compared; partial correlation analysis was used to explore the correlations of HRV parameters with PSQI scores and PSG parameters; multivariate linear regression was used to analyze the independent influencing factors for HRV.Results:(1) Compared with the ICA group, the VB group exhibited significantly higher PSQI scores, spontaneous arousal index (SAI), ratio of time of stage 1 non-rapid eye movement sleep/total sleep time (T N1/T t), and apnea-hypopnea index (AHI), while significantly lower ratio of time of rapid eye movement sleep/total sleep time (T R/T t), spindle wave density in stage 2 non-rapid eye movement sleep (N2), lowest blood oxygen saturation, standard deviation of normal to normal intervals (SDNN) of all sinus beats, low-frequency power (LF), and high-frequency power (HF, P<0.05). (2) In both VB group and ICA group, SDNN was negatively correlated with PSQI score ( r=-0.461, P=0.020; r=-0.378, P=0.036). In the VB group, SDNN was negatively correlated with T N1/T t ( r=-0.467, P=0.019) and SAI ( r=-0.551, P=0.004), and positively correlated with ratio of time of stage 3 non-rapid eye movement sleep/total sleep time (T N3/T t, r=0.686, P<0.001) and spindle wave density in N2 ( r=0.518, P=0.008); LF and HF were negatively correlated with SAI ( r=-0.481, P=0.015; r=-0.564, P=0.003). In the ICA group, HF was negatively correlated with spindle wave density in N2 ( r=-0.369; P=0.041). (3) Multivariate linear regression results indicated that T N3/T t (β=0.348, P=0.018), SAI (β=-0.330, P=0.018), and spindle wave density in N2 (β=0.286, P=0.013) were independent influencing factors for Ln_SDNN in patients with moderate-to-severe stenosis or occlusion of the vertebrobasilar system. Conclusion:Patients with stenosis or occlusion of the vertebrobasilar system exhibit poorer subjective sleep quality, increased light sleep, heightened arousal, and reduced sleep stability compared with those with stenosis or occlusion of the internal carotid artery system, which may be caused by the imbalance of autonomic nerve function.

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

Skeletal maturity can reflect an individual's developmental status and predict their future growth potential, provide clinicians with valuable diagnostic information. In recent years, significant progress has been made in imaging techniques for assessing skeletal maturity. This article aims to review the application status and research progress of X-ray, MRI and ultrasound in assessing skeletal maturity in adolescents and children, with a view of providing clinical reference.