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*

AIM: To observe the changes of corneal densitometry(CD)in patients with keratoconus after corneal cross-linking(CXL).METHODS: Retrospective study. A total of 32 patients(43 eyes)with keratoconus in Ningxia Eye Hospital from April 2020 to April 2022 were selected. Pentacam analysis system divided the cornea into three layers: anterior 120 μm, middle layer and posterior 60 μm, and divides it into five regions with diameters of 0-2, 2-6, 6-10, 10-12 mm and full diameter according to the diameter, and measures the CD in different ranges. The changes of CD were compared before operation and at 1, 3 and 6 mo after operation.RESULTS: There were differences in uncorrected visual acuity, best corrected visual acuity and intraocular pressure before and 6 mo after operation(all P<0.05), and there was no difference in corneal endothelial cells(P=0.477). CD reached its peak at 1 mo after operation, and decreased at 3 mo and 6 mo after operation, but it was still higher than that before operation. There is a significant positive correlation between CD and Kmax in the anterior layer and the whole layer(r=0.164, P=0.016; r=0.152, P=0.023).CONCLUSION: The values of CD peaked at 1 mo after CXL, then it gradually decreased, tending to become stable at 6 mo postoperatively.

OBJECTIVE To study the effects of Tongxie yaofang reducing colon hypermotility in irritable bowel syndrome with diarrhea （IBS-D） rats by regulating gut microbiota-enteric nervous system （ENS）-muscularis macrophages （MM） crosstalk. METHODS Forty newborn male SD rats were randomly divided into control group， model group， TCM group [Tongxie yaofang 2.68 g/（kg·d）， calculated by raw material]， and positive control group [Live combined bifidobacterium and lactobacillus tablets 0.27 g/（kg·d）]， with 10 rats in each group. Except for the control group， the IBS-D model of liver stagnation and spleen deficiency syndrome was induced in the other 3 groups with the method of mother-child separation+chronic restraint+Folium Sennae- induced diarrhea. After modeling， the administration groups were given relevant drug liquid intragastrically， once a day， for consecutive 2 weeks. At the end of modeling and after administration， the fecal properties （the incidence and the rate of loose stools）， colonic motility （colon emptying time）， and visceral sensitivity [abdominal withdrawal reflex （AWR） scores under different pressures] of rats were observed in each group. The concentration of lipopolysaccharide （LPS） in serum was detected after medication， and the expressions and distribution of bone morphogenetic protein 2 （BMP2）， colony stimulating factor 1 （CSF1） and Toll-like receptor 4 （TLR4） in colon tissue were detected； the diversity， species composition and differences of gut microbiota were also determined. RESULTS At the end of modeling， compared with the control group， all rats of the other three groups suffered from loose stools （100%）， the rate of loose stools and AWR scores at different pressures increased significantly， and colon emptying time was shortened significantly （P＜0.01 or P＜0.05）. The incidences of loose stools were 20% in TCM group and 80% in the positive control group； the rate of loose stools and AWR scores at different pressures， serum concentration of LPS and protein expressions of CSF1 and TLR4 in muscle layer of colon tissue in TCM group significantly decreased， compared with the model group； colon emptying time， the average optical density of BMP2 protein in muscle layer of colon tissue， and the indexes of Chao 1， Shannon and Faith’s PD and Simpson E-mail：772699670@qq.com index of rats in TCM group were all prolonged or increased significantly， compared with the model group （P＜0.01 or P＜ E-mail：aiwangzi312@163.com 0.05）. The relative abundance ratio of Firmicutes/Bacteroidetes， from low to high， was in the order of TCM group， control group， positive control group and model group； the species composition of gut microbiota in TCM group was closer to control group， with dominant bacterial genera including Prevotella and Blautia. CONCLUSIONS Tongxie yaofang can regulate the expressions of BMP2 and CSF1， the key proteins of gut microbiota-ENS-MM crosstalk， by changing the gut microbiota， thus alleviating the abnormal hyperfunction of colon motility in IBS-D rats.

BACKGROUND:Obesity has become a global health issue,often accompanied by complications including obesity-related muscle atrophy.While exercise has been reported to improve various obesity-related diseases,there is limited research focusing on exercise modes. OBJECTIVE:To compare the effects of moderate-intensity continuous training(MICT)and high-intensity interval training(HIIT)on obesity-related muscle atrophy in mice under the premise of the same exercise distance,providing a scientific basis for exercise interventions for obesity-related muscle atrophy. METHODS:Seventy-two male C57BL/6 mice were divided into six groups(n=12 per group):standard chow diet,standard chow diet+MICT,standard chow diet+HIIT,high-fat diet,high-fat diet+MICT,and high-fat diet+HIIT.The study evaluated the effects of 8-week treadmill training with different exercise modes on long-term high-fat diet-induced muscle atrophy by detecting muscle mass,muscle index,muscle fiber cross-sectional area,muscle lipid deposition,and the expression of muscle atrophy marker genes Murf-1 and Atrogin-1 in the gastrocnemius muscle of mice exposed to long-term high-fat diet. RESULTS AND CONCLUSION:Compared to the high-fat diet group,both MICT and HIIT improved the decrease in gastrocnemius muscle index(MICT+18.8%vs.HIIT+17.6%,not significant between the two modes),muscle fiber atrophy(MICT+15.5%vs.HIIT+13.7%,not significant between the two modes),and muscle lipid deposition(MICT-19.8%vs.HIIT-17.1%,not significant between the two modes).At the gene level,compared with the high-fat diet group,both MICT and HIIT could significantly down-regulate the expression of Murf-1(MICT-62.4%vs.HIIT-52.6%,the down-regulation caused by MICT was significantly greater than that by HIIT;P＜0.01)and Atrogin-1(MICT-43.3%vs.HIIT-29.8%,the down-regulation caused by MICT was significantly greater than that by HIIT;P＜0.01).Based on exercise mode comfort and genetic evidence,MICT mode might be more suitable for exercise interventions in obesity-related muscle atrophy.

Mucin-type O-glycosylation is one of the most common post-translational modifications in proteins,capable of altering protein conformation and biological functions.It plays a crucial role in biological processes such as cell signaling,cell adhesion,and immune responses.Polypeptide N-acetylgalactosaminyltransferase 3(GALNT3),as the initiating enzyme of mucin-type O-glycosylation,is of paramount importance in maintaining the homeostasis of human cells and tissues.Dysfunction of GALNT3 has been found to play a role in various diseases,such as calcium-phosphorus metabolism disorders and atherosclerosis.Additionally,GALNT3 is abnormally expressed in several types of tumors,including colorectal cancer,lung cancer,and ovarian cancer.Its expression is associated with the clinical pathological features of patients and poor prognosis,making it a potential biomarker for early tumor diagnosis and prognosis evaluation.Further research shows that GALNT3 can both regulate glycosylation levels to reduce adhesion between tumor cells and activate multiple metabolism-related pathways,promoting tumor cell invasion and metastasis.This review summarizes the role of GALNT3 in the development of malignant tumors and discusses the prospects and challenges of developing anti-tumor drugs targeting GALNT3.

Objective:To investigate the microsurgical efficacy of large primary intracranial solitary fibrous tumor and influencing factors for its prognoses.Methods:From January 2010 to December 2022, 47 patients with large primary intracranial solitary fibrous tumor admitted to and accepted microsurgery in Department of Neurosurgery, Wuhan Central Hospital and Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, were enrolled. The clinical data, microsurgical efficacy and follow-up results of these patients were retrospectively analyzed, and influencing factors for total resection and prognoses were determined.Results:Thirty-two patients had tumor within the supratentorial region and 15 in the mandibular region, including 24 with sinus involved tumor. According to 2021 WHO Classification of Tumors of the Central Nervous System, 5 patients (10.6%) had grading 1, 32 (68.1%) grading 2, and 10 (21.3%) grading 3. Total resection was achieved in 31 patients (66.0%) and subtotal resection in 16 patients (34.0%). Postoperative complications, such as intraoperative hemorrhage, distant epidural hematoma and subcutaneous effusion, occurred in 7 patients (14.9%) and they were cured after secondary hematoma removal or conservative treatment; residual limb mobility disorder occurred in 3 patients, visual impairment in 3, and postoperative seizures in 2. Adjuvant radiotherapy was performed in 13 patients (27.7%). Follow-up was performed for (69.1±29.6) months and 29 patients (61.7%) had recurrent tumors (6 with intracranial and extracranial metastases and 4 deaths). Mean progression-free survival was (57.5±25.1) months; the 1-, 3-, and 5-year progression-free survival rates were 95.7%, 87.2%, and 59.6%, respectively. Sinus involvement was the independent influencing factor for total tumor resection; and total resection was an independent protective factor for progression-free survival for large primary intracranial solitary fibrous tumor ( HR=4.291, 95% CI: 1.555-11.838, P=0.005). Conclusion:Patients with large primary intracranial solitary fibrous tumor have a high recurrent risk after surgery; and gross-total resection should be strived to prevent tumor recurrence.

Objective Screening of modified Foshou San to improve cerebral ischemia,determine the best prescription composition and the proper dose.Methods Using data mining method,the traditional Chinese medicine for hypertensive intracerebral hemorrhage(HICH)were screened,and cluster analysis was carried out to obtain Jiawei Foshou San combinations;According to the mixed uniform design U14(42×33×22),the auricular artery dilatation experiment and acute cerebral ischemia induced by saturated magnesium chloride in mice were used to observe the auricular artery dilatation index and survival time of mice,and the pharmacological effects of each prescription to improve cerebral ischemia were evaluated.On the basis of visual analysis of pharmacodynamics,stepwise multiple regression analysis was carried out,and the regression equation was used to calculate the best prescription composition and dose of Jiawei Foshou San.The optimized prescription was verified by acute cerebral ischemia experiment in mice.Results A total of 51 prescriptions that may be used for the treatment of HICH were selected by data mining,including 103 Chinese herbs,with a total frequency of 535 times.The results of mixed uniform design U14(42×33×22)test showed that the auricular artery index of mice in each prescription group of Jiawei Foshou San increased to varying degrees.The differences between prescription 4,8,12 and 13 were significant(P<0.05),and the differences between prescription 3,7,9,11 and 14 were extremely significant(P<0.01).The survival time of acute cerebral ischemia in mice was prolonged to varying degrees in each group of Jiawei Foshou San,and the difference between prescription 2-14 groups and the blank group was statistically significant(P<0.01).The prescription of the new Jiawei Foshou San was determined as 60 g of Astragalus membranaceus,60 g of Angelica sinensis,24 g of Ligusticum chuanxiong,15 g of Paeonia lactiflora and 9 g of Dilong from the intuitive analysis.The validation results showed that the high and middle dosage of the new Jiawei Foshou San could significantly prolong the breathing time,increase the number of breaths,and prolong the time of normal pressure hypoxia tolerance in mice with acute cerebral ischemia(P<0.01,P<0.05).Conclusion Using mixed uniform design combined with improving cerebral ischemia effect index can determine the best prescription composition and the best dose of Jiawei Foshou San,and the effect of improving cerebral ischemia is certain.

Objective:To explore the effect of ultra-early nursing intervention on postoperative cognitive behavior and prognosis of patients with cerebral hemorrhage.Methods:Using the convenient sampling method, a total of 82 patients with cerebral hemorrhage who were admitted to the First Affiliated Hospital of Zhengzhou University from March 2019 to January 2021 were selected as the research objects. According to the random number table method, they were divided into the control group and the observation group, with 41 cases in each group. Both groups received surgical treatment. The control group received routine nursing, while the observation group received ultra-early nursing intervention on the basis of routine nursing when vital signs were stable 48 hours after operation. The differences in neurological function (National Institutes of Health Stroke Scale, NIHSS) , cognitive function (Loewenstein Occupational Therapy Cognitive Assessment, LOTCA) and activity of daily living scores were compared between the two groups at 48 hours and 1 month after operation.Results:The NIHSS score at 1 month after surgery in both groups was lower than that at 48 hours after surgery and the NIHSS score at 1 month after surgery in observation group was lower than that in control group ( P<0.05) . The LOTCA dimension scores of the two groups at 1 month after operation were higher than those at 48 hours after operation and the LOTCA dimension scores of the observation group at 1 month after operation were higher than those of the control group ( P<0.05) . The activity of daily living scores of the two groups at 1 month after operation were higher than those at 48 hours after operation and the activity of daily living score of the observation group at 1 month after operation was higher than that of the control group , the differences were statistically significant ( P<0.05) . Conclusions:Ultra-early nursing intervention can effectively promote the recovery of postoperative neurological function and cognitive function in patients with cerebral hemorrhage and improve the activity of daily living of patients.