To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang （SWDK） on cognitive dysfunction in mice suffering from Alzheimer's disease （AD） and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group， total ginsenoside group（0.04 g·kg^-1）， high-， medium-， and low-dose groups of SWDK （32.60， 16.30， 8.15 g·kg^-1）， with 10 mice in each group， and ten wild-type mice of the same age were used as the normal group， male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin （HE） staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin （SYP） and postsynaptic dense substance 95 （PSD95） in mice serum were detected by enzyme-linked immunosorbent assay （ELISA）. The positive expression of brain-derived neurotrophic factor（BDNF） in the dentate gyrus （DG） region of mouse brain tissue was observed by immunohistochemistry （IHC）. The protein levels of BDNF， Wnt family member 3A（Wnt3a）， and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice， the model group of mice had significantly more complex swimming routes and lower swimming speed （P<0.01）， significantly lower percentage of time spent in the target quadrant （P<0.01）， and a significantly lower percentage of freezing time （P<0.05）. The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95（P<0.01） in the serum of mice were reduced， and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice were obviously reduced （P<0.05， P<0.01）. Compared with the model group， the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes， the high- and medium- dose SWDK groups significantly higher swimming speeds （P<0.01）， significantly higher percentage of time spent in the target quadrant （P<0.01）， obviously higher percentage of Freezing time （P<0.05）， and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP （P<0.05， P<0.01）， PSD95 （P<0.01）， increased BDNF-positive cells in the DG region of brain tissue， and obviously elevated levels of BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice （P<0.05， P<0.01）. ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice， and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway， which promotes BDNF expression and thereby enhances synaptic plasticity， allowing neuronal signaling to be restored.

ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang （SWDK） on cognitive dysfunction in mice suffering from Alzheimer's disease （AD） and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group， total ginsenoside group（0.04 g·kg^-1）， high-， medium-， and low-dose groups of SWDK （32.60， 16.30， 8.15 g·kg^-1）， with 10 mice in each group， and ten wild-type mice of the same age were used as the normal group， male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin （HE） staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin （SYP） and postsynaptic dense substance 95 （PSD95） in mice serum were detected by enzyme-linked immunosorbent assay （ELISA）. The positive expression of brain-derived neurotrophic factor（BDNF） in the dentate gyrus （DG） region of mouse brain tissue was observed by immunohistochemistry （IHC）. The protein levels of BDNF， Wnt family member 3A（Wnt3a）， and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice， the model group of mice had significantly more complex swimming routes and lower swimming speed （P<0.01）， significantly lower percentage of time spent in the target quadrant （P<0.01）， and a significantly lower percentage of freezing time （P<0.05）. The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95（P<0.01） in the serum of mice were reduced， and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice were obviously reduced （P<0.05， P<0.01）. Compared with the model group， the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes， the high- and medium- dose SWDK groups significantly higher swimming speeds （P<0.01）， significantly higher percentage of time spent in the target quadrant （P<0.01）， obviously higher percentage of Freezing time （P<0.05）， and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP （P<0.05， P<0.01）， PSD95 （P<0.01）， increased BDNF-positive cells in the DG region of brain tissue， and obviously elevated levels of BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice （P<0.05， P<0.01）. ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice， and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway， which promotes BDNF expression and thereby enhances synaptic plasticity， allowing neuronal signaling to be restored.

Objective:To compare the clinical effects of pedicle screw internal fixation under different approaches for the treatment of asymptomatic thoracolumbar fractures.Methods:Retrospective analysis was conducted to collect data from 83 patients with asymptomatic thoracolumbar fractures treated with pedicle screw internal fixation at Handan Central Hospital from January 2023 to December 2023. Based on different surgical approaches, 40 patients who underwent regular percutaneous approach surgery were included in the conventional approach group, and 43 patients who underwent modified Wiltse approach surgery were included in the modified approach group. The perioperative indexes and postoperative complications of the two groups were statistically analyzed. The visual analogue score (VAS) was compared before operation, 1 month after operation and 3 months after operation. The anterior vertebral height, Cobb angle and fracture healing factors were compared before operation and 3 months after operation.Results:The surgical time, incision length and intraoperative fluoroscopy frequency of the modified approach group were all lower than those of the conventional approach group: (65.75 ± 7.16) min vs. (74.28 ± 6.85) min, (6.75 ± 0.42) cm vs. (9.58 ± 0.50) cm and (9.22 ± 1.24) times vs. (21.03 ± 2.45) times, with statistically significant differences ( P<0.05). The total incidence of postoperative complications in the modified approach group was 9.30% (4/43), while the total incidence of postoperative complications in the conventional approach group was 15.00% (6/40), with no statistically significant difference ( P>0.05). The VAS of the modified approach group was lower than that of the conventional approach group at 1 and 3 months after surgery: (3.05 ± 0.82) points vs. (3.86 ± 0.85) points and (1.62 ± 0.32) points vs. (2.24 ± 0.54) points, with statistically significant differences ( P<0.05). Three months after surgery, the height of the anterior vertebral edge, serum type Ⅰ procollagen N-terminal propeptide and platelet-derived growth factor in the modified approach group were significantly higher than those in the conventional approach group: (72.96 ± 9.27)% vs. (67.58 ± 9.02)%, (174.36 ± 16.75) μg/L vs. (146.24 ± 15.86) μg/L, (250.28 ± 14.62) U/L vs. (241.20 ± 12.75) U/L, the Cobb angle was lower than that of the conventional approach group: (10.15 ± 1.32)° vs. (12.20 ± 1.86)°, with statistically significant differences ( P<0.05). Conclusions:The modified Wiltse approach and percutaneous approach pedicle screw internal fixation are safe and effective in treating asymptomatic thoracolumbar fractures; but compared to the latter, the former has a shorter surgical time, fewer intraoperative fluoroscopy sessions, and less postoperative pain. It has better effects in improving the height of the anterior edge of the injured vertebra, Cobb angle recovery, and fracture healing factors.

Objective:To compare the clinical effects of pedicle screw internal fixation under different approaches for the treatment of asymptomatic thoracolumbar fractures.Methods:Retrospective analysis was conducted to collect data from 83 patients with asymptomatic thoracolumbar fractures treated with pedicle screw internal fixation at Handan Central Hospital from January 2023 to December 2023. Based on different surgical approaches, 40 patients who underwent regular percutaneous approach surgery were included in the conventional approach group, and 43 patients who underwent modified Wiltse approach surgery were included in the modified approach group. The perioperative indexes and postoperative complications of the two groups were statistically analyzed. The visual analogue score (VAS) was compared before operation, 1 month after operation and 3 months after operation. The anterior vertebral height, Cobb angle and fracture healing factors were compared before operation and 3 months after operation.Results:The surgical time, incision length and intraoperative fluoroscopy frequency of the modified approach group were all lower than those of the conventional approach group: (65.75 ± 7.16) min vs. (74.28 ± 6.85) min, (6.75 ± 0.42) cm vs. (9.58 ± 0.50) cm and (9.22 ± 1.24) times vs. (21.03 ± 2.45) times, with statistically significant differences ( P<0.05). The total incidence of postoperative complications in the modified approach group was 9.30% (4/43), while the total incidence of postoperative complications in the conventional approach group was 15.00% (6/40), with no statistically significant difference ( P>0.05). The VAS of the modified approach group was lower than that of the conventional approach group at 1 and 3 months after surgery: (3.05 ± 0.82) points vs. (3.86 ± 0.85) points and (1.62 ± 0.32) points vs. (2.24 ± 0.54) points, with statistically significant differences ( P<0.05). Three months after surgery, the height of the anterior vertebral edge, serum type Ⅰ procollagen N-terminal propeptide and platelet-derived growth factor in the modified approach group were significantly higher than those in the conventional approach group: (72.96 ± 9.27)% vs. (67.58 ± 9.02)%, (174.36 ± 16.75) μg/L vs. (146.24 ± 15.86) μg/L, (250.28 ± 14.62) U/L vs. (241.20 ± 12.75) U/L, the Cobb angle was lower than that of the conventional approach group: (10.15 ± 1.32)° vs. (12.20 ± 1.86)°, with statistically significant differences ( P<0.05). Conclusions:The modified Wiltse approach and percutaneous approach pedicle screw internal fixation are safe and effective in treating asymptomatic thoracolumbar fractures; but compared to the latter, the former has a shorter surgical time, fewer intraoperative fluoroscopy sessions, and less postoperative pain. It has better effects in improving the height of the anterior edge of the injured vertebra, Cobb angle recovery, and fracture healing factors.

Objective To investigate the role of immunoglobulin superfamily containing leucine-rich repeat protein(ISLR)in the malignant progression of osteosarcoma cells and its potential regulatory mechanism.Methods ISLR mRNA levels in osteosarcoma tissues and cells were detected by quantitative real time polymerase chain reaction(qRT-PCR).U2OS cells were transfected with ISLR short hairpin RNA(shRNA)sequence or negative-control shRNA(NC shRNA)sequence,thus the cells were treated with phosphatidylinositol 3 kinase(PI3K)activator 740 Y-P.The cell viability,invasion ability and apoptosis rate were detected by CCK-8 assay,Transwell assay and flow cytometry,respectively.Western blot was used to detect the expressions of ISLR protein,epithelial-mesenchymal transition(EMT)-related proteins[Epitheia-cadherin(E-cadherin),Nerve cadherin(N-cadherin),Vimentin,Snail],PI3K/protein kinase B(AKT)pathline-related proteins,apoptotic proteins[Cysteinyl aspartate-specific proteinase-3(Caspase-3),B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax)]and proliferation marker Ki67 protein.Lentivirus was used to transfect U2OS cells,and the cells were injected into nude mice to construct a xenograft tumor model,and tumor growth was monitored.Results ISLR mRNA level in osteosarcoma tissue(5.14±1.63)was up-regulated compared with para-cancerous tissue(1.01±0.02),and the difference was significant(t=-14.332,P＜0.001).Compared with normal osteoblasts hFOB1.19(1.01±0.01),osteosarcoma cells MG63(3.05±0.57),U2OS(4.55±0.79),HOS(2.46±0.41),the relative expression of ISLR mRNA in Saos-2(2.62±0.44)and 143B(3.62±0.51)were increased,and differences were significant(t=4.883,8.473,3.471,3.854,6.247,all P＜0.05).Silencing ISLR inhibited the proliferation of U2OS cells(t=6.593,6.835)and invasion(t=8.621,8.448),but promoted cell apoptosis(t=25.505,25.574),and the differences were significant(all P＜0.05).Silencing ISLR promoted Caspase-3 activity in U2OS cells(t=13.489,13.366)and Bax protein(t=8.628,8.524),but inhibited Bcl-2 protein expression(t=10.948,10.775),with significant differences(all P＜0.05).Silencing ISLR promoted EMT-related protein E-cadherin(t=15.168,15.087),inhibited N-cadherin(t=10.220,10.058),Vimentin(t=8.303,8.164)and Snail(t=9.211,9.384),but reduced the phosphorylation levels of PI3K and AKT(t=17.441,14.452),with significant differences(all P＜0.05).Additionally,740 Y-P treatment reversed the effect of silencing ISLR on U2OS cells.Experimental results in vivo showed that knockdown of ISLR significantly inhibited tumor growth.Conclusion ISLR could promote EMT,proliferation and invasion,but inhibit apoptosis of osteosarcoma cells by activating the PI3K/AKT pathway,there by promoting osteosarcoma progression.

Objective: To understand the distribution of tobacco retailer within 100 meters outside middle schools in Wuhan City and its impact on smoking behavior of middle school students, so as to provide basis and feasible suggestions for the development of tobacco control policy for adolescents. Methods: From February to May 2023, a multi stage stratified cluster random sampling method was used to select 20 middle schools from 4 districts in Wuhan City. To investigate the distribution of tobacco retailer within 100 metres outside the school and the sale of tobacco to minors. A total of 4 882 students were surveyed using the core questions of the 2021 Chinese Adolescent Tobacco Prevalence Questionnaire. Fisher exact probability test, Chi square test and Chi square trend test were used for statistical analysis. Results: Nearly 70.00% of middle schools had tobacco retailer within 100 metres, with an average of (1.10±0.97) per middle school. The awareness rate (100.00%) and labeling rate (87.50%) of licensed tobacco retailer were higher than those of non licensed tobacco retailer (33.33%, 16.67%) ( P <0.05). The rates of tried smoking, current smoking and buying cigarettes within 30 days were 7.13%, 1.99% and 2.54%, respectively. The rates of students who tried smoking ( 8.58 %), current smoking (2.29%) and buying cigarettes within 30 days (2.85%) in schools with tobacco retailer within 100 metres were higher than those in schools without tobacco retailer (3.79%, 1.28%, 1.83%)( χ 2=35.80, 5.37, 4.37 , P <0.05). And as the grade increased, the rates of tried smoking, current smoking and buying cigarettes among middle school students all showed an upward trend ( χ 2 trend =66.20, 36.10, 16.17, P <0.05). Conclusions Middle school students in Wuhan City have high tobacco availability. The findings suggest that school ban should be extended from 50 meters to 100 meters, and the regulatory authorities must strictly prohibit selling tobacco products to minors at tobacco retailer.

During coronavirus disease 2019 epidemic, the treatment of severe trauma has been impacted. The Consensus on emergency surgery and infection prevention and control for severe trauma patients with 2019 novel corona virus pneumonia was published online on February 12, 2020, providing a strong guidance for the emergency treatment of severe trauma and the self-protection of medical staffs in the early stage of the epidemic. With the Joint Prevention and Control Mechanism of the State Council renaming "novel coronavirus pneumonia" to "novel coronavirus infection" and the infection being managed with measures against class B infectious diseases since January 8, 2023, the consensus published in 2020 is no longer applicable to the emergency treatment of severe trauma in the new stage of epidemic prevention and control. In this context, led by the Chinese Traumatology Association, Chinese Trauma Surgeon Association, Trauma Medicine Branch of Chinese International Exchange and Promotive Association for Medical and Health Care, and Editorial Board of Chinese Journal of Traumatology, the Chinese expert consensus on emergency surgery for severe trauma and infection prevention during coronavirus disease 2019 epidemic ( version 2023) is formulated to ensure the effectiveness and safety in the treatment of severe trauma in the new stage. Based on the policy of the Joint Prevention and Control Mechanism of the State Council and by using evidence-based medical evidence as well as Delphi expert consultation and voting, 16 recommendations are put forward from the four aspects of the related definitions, infection prevention, preoperative assessment and preparation, emergency operation and postoperative management, hoping to provide a reference for severe trauma care in the new stage of the epidemic prevention and control.

Objective:To explore the clinical application of portable head and neck magnetic resonance imaging (MRI) device in neurosurgery.Methods:A total of 213 patients with brain diseases who were scanned by portable head and neck MRI device in Center of Neurosurgery, Zhujiang Hospital, Southern Medical University from June to September 2022 were selected. The portable head and neck MRI images and 3.0T conventional MRI images of 10 randomly selected patients were compared; the differences in signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of different sequences were analyzed. Thirty-one patients accepted tracheal intubation/tracheotomy, or ventilator-assisted breathing were selected as special patient group, and another 30 patients were as general patient group; the differences in comprehensive diagnostic scores of portable head and neck MRI images were compared. Noise intensity differences in different sequences between 3.0T conventional MRI and portable head and neck MRI were statistically compared. Twenty hospitalized volunteers with normal hearing in our center from July to August 2022 were selected, conventional 3.0T MRI and portable head and neck MRI were performed successively, and the noise intensity of different sequences in them was evaluated by using a 5-point system.Results:Compared with those in 3.0T conventional MRI images, the SNR and CNR of T1WI, T2WI, and Liquid attenuated reverse recovery sequence (FLAIR) sequences in portable head and neck MRI images were significantly lower ( P<0.05). No significant difference was noted in the comprehensive diagnostic scores of portable head and neck MRI images between special patients and general patients ( P>0.05). Compared with that in the 3.0T conventional MRI, the noise intensity of different sequences in portable head and neck MRI was significantly reduced ( P<0.05). These volunteers had significantly reduced noise intensity scores of different sequences in portable head and neck MRI compared with that in conventional 3.0T MRI ( P<0.05). Conclusion:Portable head and neck MRI device is easy to use, enjoying high safety, imaging quality and suitability, which meets the clinical needs for neurosurgery patients.