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 discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

Objective:To investigate the effects of Yangrong Changchun Ointment on serum estradiol (E 2), follicle stimulating hormone (FSH), anti-Müllerian hormone (AMH), and on the Kiss peptin/protein one anti-G protein-coupled receptor 54 (GPR54) signaling pathway in rats with menopause syndrome (MPS). Methods:Totally 48 SD female rats were randomly divided into a sham-operation group ( n=8) and a modeling group ( n=40), and the rat model of menopausal syndrome was established by ovariectomy. After successful modeling, the rats were divided into the model group, Kunbao Pills group group, Yangrong Changchun Ointment high-, medium-, low-dosage groups according to random number table method, with eight rats in each group. Yangrong Changchun Ointment high-, medium-, low-dosage groups were orally administered with Yangrong Changchun Ointment suspension at doses of 10.0, 5.0, and 2.5 g/kg, while the Kunbao Pills group was orally administered with Kunbao Pills suspension at a dosage of 1 g/kg. The sham-operation group and model group were orally administered with distilled water of equal volume at a dosage of 1 ml/100 g body weight, once a day, for 28 consecutive days. The organ index of rats was calculated; the behavior of rats was determined by open filed test; HE staining was used to observe the histopathology of the uterus; ELISA was used to detect the levels of E 2, FSH, and AMH in the serum of rats; and qRT-PCR and Western blot were used to detect the mRNA and protein expressions of GPR54, Kiss-1, Phospholipase C (PLC), gonadotropin-releasing hormone (GnRH), and protein kinase C (PKC) in the hypothalamus. Results:Compared with the model group, the uterus, adrenal gland, spleen and thymus indices increased in the Yangrong Changchun Ointment high-dosage group ( P<0.05); serum levels of E 2 and AMH increased ( P<0.05), and the serum level of FSH decreased ( P<0.05); the mRNA and protein expressions of GPR54, Kiss-1, PLC, GnRH and PKC in the hypothalamus decreased ( P<0.05), and the uterine histopathological damage was improved. Conclusion:Yangrong Changchun Ointment can improve the pathological injury and estrogen level disorder in rats with MPS, and its mechanism may be related to inhibiting the activation of kisspeptin/GPR54 pathway.

Battalion aid stations usually serve as the critical bridge between medical rescue and early treatment,whose treatment efficiency will make a huge difference in the survival rate of casualties on battlefields in the future.This paper reviews the evolution of the functions of U.S.military battalion aid stations.By taking the U.S.Marine Corps and the 75th Ranger Regiment,this article elaborates on the practices by the U.S.military in the construction of battalion aid stations,and unveils the takeaways from the construction of the army battalion aid stations and their limitations.It is recommended that future battalion aid stations should set support tasks based on the need of medical services,optimize the make-up of battalion aid stations,improve their support capabilities and informatization,and provide better en-route treatment in order to ensure battalion aid stations can play a more important role.

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.

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.

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.