BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

Objective To propose a novel method based on three-dimensional depthwise separable convolution network(3D DSN)for the separation of PET images with dual tracers of 18F-FDG and 18F-FAPI.Methods A total of 120 pairs of 18F-FDG and 18F-FAPI PET images of the same patient scanned separately at different time points were collected,and the dual-tracer PET image was generated through simulation.After the image registration of PET images of two tracers for ensuring spatial position matching,the registered PET images were forward-projected to generate sinogram data,and the sinogram data of two tracers were accumulated to obtain mixed sinogram data.Subsequently,the dual-tracer PET image was reconstructed using maximum likelihood expectation maximization and input into a 3D DSN based network for image separation,thereby obtaining PET images of two single tracers.Results Compared with 3D CNN method,the proposed method increased the structure similarity index measure(SSIM)of the separated 18F-FDG images to the real 18F-FDG images by 0.87%,increased the peak signal-to-noise ratio(PSNR)by 11.8%,and reduced the normalized root mean square error(NRMSE)by 52%.The SSIM of the separated 18F-FAPI images to the real 18F-FAPI images increased by 1.1%,PSNR increased by 17.0%,and NRMSE decreased by 51%.Conclusion The proposed method can be effectively applied to simultaneous PET imaging with dual PET tracers,reducing the number of scans and costs in time and money,and providing clinical doctors more accurate and abundant diagnostic information.

BACKGROUND:Previous studies have indicated that cathepsin K can intervene with the occurrence and development of osteoporosis by regulating bone mineral density in middle-aged and older adults. However,whether there is a causal relationship between the cathepsin family and bone mineral density in other populations remains unknown. OBJECTIVE:To investigate the causal relationship between cathepsin and bone mineral density.METHODS:Genetic loci associated with eight cathepins were extracted from the IEU Open GWAS database as instrumental variables,and bone mineral density values in five age groups acted as an outcome. The causal relationship between cathepin and bone mineral density was assessed by two-way Mendelian randomization analysis. Heterogeneity of the genetic instrumental variables was assessed using Cochran's Q test,pleiotropy was assessed using the MR-Egger intercept test,and the sensitivity of single nucleotide polymorphisms used as instrumental variables to the causal effect of exposure and outcome was assessed using the leave-one-out method. RESULTS AND CONCLUSION:The results of the inverse variance weighting method with positive Mendelian randomization showed that cathepin H was negatively associated with bone mineral density in people aged 45-60 years[odds ratio (95％ confidence interval)=0.965(0.94-0.99),P=0.04];cathepin Z was negatively associated with bone mineral density in people aged 30-45 year[odds ratio (95％ confidence interval)=1.06 (1.00-1.11),P=0.03]. The results of sensitivity analysis showed a stable causal relationship,and MR-Egger intercept analysis did not detect potential horizontal pleiotropy. The inverse Mendelian randomization results showed that bone mineral density had no significant inverse effect on cathepin. The above results confirm that cathepin can affect bone mineral density in some age groups,which may increase the risk of osteoporosis and should be given more attention.

Objective To investigate the expression levels of Ubiquitin Carboxy-Terminal Hydrolase-L1(UCH-L1)and Netrin-1 in the serum of patients with spontaneous basal ganglia hemorrhage(ICH)complicated with cerebral edema,and to analyze their impacts on neurological deficits and prognosis.Methods A retrospec-tive analysis was conducted on the clinical data of 173 patients with spontaneous basal ganglia hemorrhage admitted to the Department of Neurosurgery,The First Affiliated Hospital of Bengbu Medical University from September 2023 to January 2025.The serum levels of UCH-L1 and Netrin-1 were measured within 24 hours after the onset.They were divided into three groups according to the size of the cerebral edema volume(CEV):Group A(CEV<10 mL),Group B(CEV 10～30 mL),and Group C(CEV>30 mL).Pearson's correlation analysis was used to analyze the correlation between serum expression levels of UCH-L1 and Netrin-1 with hemorrhage,volume of cere-bral edema,distance of midline shift,Modified Edinburgh-Scandinavian Stroke Scale(MESSS)score,Modified Rankin Scale(mRS)score,and Glasgow Coma Scale(GCS)score.Logistic regression analysis was performed to identify the risk factors for poor prognosis.Receiver operating characteristic(ROC)curve analysis was used to evaluate the predictive value of UCH-L1 and netrin-1 for poor prognosis.Results Significant differences were observed in the serum levels of UCH-L1 and netrin-1 among patients with different volumes of cerebral edema(P<0.05).The larger the volume of cerebral edema,the higher the expression levels of UCH-L1 and netrin-1.The serum levels of UCH-L1 and Netrin-1 were significantly higher in the poor prognosis group compared to the good prognosis group(P<0.05).The serum levels of UCH-L1 and Netrin-1 were positively correlated with MESSS score,hemorrhage volume,cerebral edema volume,distance of midline shift,and mRS score(P<0.05),and negatively correlated with GCS score(P<0.05).Multivariate Logistic regression analysis showed that both UCH-L1 and netrin-1 were independent risk factors for poor neurological prognosis in basal ganglia hemorrhage patients(P<0.05).ROC curve analysis indicated that both markers had important predictive value for poor prognosis.The AUC for serum UCH-L1 level predicting poor prognosis was 0.77[95%confidence interval(CI):0.69～0.85,P<0.01],with a sensitivity of 84.9%and a specificity of 50.6%.The AUC for serum Netrin-1 level predicting poor prognosis was 0.89(95%CI:0.85～0.94,P<0.01),with a sensitivity of 82.1%and a specificity of 68.7%.Conclusions Serum UCH-L1 and Netrin-1 are differentially expressed in patients with spontaneous basal ganglia hemorrhage complicated with different volumes of cerebral edema.They are independent risk factors for poor prog-nosis and are important predictors of neurological function prognosis in these patients.

BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

Objective To investigate the expression levels of Ubiquitin Carboxy-Terminal Hydrolase-L1(UCH-L1)and Netrin-1 in the serum of patients with spontaneous basal ganglia hemorrhage(ICH)complicated with cerebral edema,and to analyze their impacts on neurological deficits and prognosis.Methods A retrospec-tive analysis was conducted on the clinical data of 173 patients with spontaneous basal ganglia hemorrhage admitted to the Department of Neurosurgery,The First Affiliated Hospital of Bengbu Medical University from September 2023 to January 2025.The serum levels of UCH-L1 and Netrin-1 were measured within 24 hours after the onset.They were divided into three groups according to the size of the cerebral edema volume(CEV):Group A(CEV<10 mL),Group B(CEV 10～30 mL),and Group C(CEV>30 mL).Pearson's correlation analysis was used to analyze the correlation between serum expression levels of UCH-L1 and Netrin-1 with hemorrhage,volume of cere-bral edema,distance of midline shift,Modified Edinburgh-Scandinavian Stroke Scale(MESSS)score,Modified Rankin Scale(mRS)score,and Glasgow Coma Scale(GCS)score.Logistic regression analysis was performed to identify the risk factors for poor prognosis.Receiver operating characteristic(ROC)curve analysis was used to evaluate the predictive value of UCH-L1 and netrin-1 for poor prognosis.Results Significant differences were observed in the serum levels of UCH-L1 and netrin-1 among patients with different volumes of cerebral edema(P<0.05).The larger the volume of cerebral edema,the higher the expression levels of UCH-L1 and netrin-1.The serum levels of UCH-L1 and Netrin-1 were significantly higher in the poor prognosis group compared to the good prognosis group(P<0.05).The serum levels of UCH-L1 and Netrin-1 were positively correlated with MESSS score,hemorrhage volume,cerebral edema volume,distance of midline shift,and mRS score(P<0.05),and negatively correlated with GCS score(P<0.05).Multivariate Logistic regression analysis showed that both UCH-L1 and netrin-1 were independent risk factors for poor neurological prognosis in basal ganglia hemorrhage patients(P<0.05).ROC curve analysis indicated that both markers had important predictive value for poor prognosis.The AUC for serum UCH-L1 level predicting poor prognosis was 0.77[95%confidence interval(CI):0.69～0.85,P<0.01],with a sensitivity of 84.9%and a specificity of 50.6%.The AUC for serum Netrin-1 level predicting poor prognosis was 0.89(95%CI:0.85～0.94,P<0.01),with a sensitivity of 82.1%and a specificity of 68.7%.Conclusions Serum UCH-L1 and Netrin-1 are differentially expressed in patients with spontaneous basal ganglia hemorrhage complicated with different volumes of cerebral edema.They are independent risk factors for poor prog-nosis and are important predictors of neurological function prognosis in these patients.

Objective To propose a novel method based on three-dimensional depthwise separable convolution network(3D DSN)for the separation of PET images with dual tracers of 18F-FDG and 18F-FAPI.Methods A total of 120 pairs of 18F-FDG and 18F-FAPI PET images of the same patient scanned separately at different time points were collected,and the dual-tracer PET image was generated through simulation.After the image registration of PET images of two tracers for ensuring spatial position matching,the registered PET images were forward-projected to generate sinogram data,and the sinogram data of two tracers were accumulated to obtain mixed sinogram data.Subsequently,the dual-tracer PET image was reconstructed using maximum likelihood expectation maximization and input into a 3D DSN based network for image separation,thereby obtaining PET images of two single tracers.Results Compared with 3D CNN method,the proposed method increased the structure similarity index measure(SSIM)of the separated 18F-FDG images to the real 18F-FDG images by 0.87%,increased the peak signal-to-noise ratio(PSNR)by 11.8%,and reduced the normalized root mean square error(NRMSE)by 52%.The SSIM of the separated 18F-FAPI images to the real 18F-FAPI images increased by 1.1%,PSNR increased by 17.0%,and NRMSE decreased by 51%.Conclusion The proposed method can be effectively applied to simultaneous PET imaging with dual PET tracers,reducing the number of scans and costs in time and money,and providing clinical doctors more accurate and abundant diagnostic information.

BACKGROUND:Previous studies have indicated that cathepsin K can intervene with the occurrence and development of osteoporosis by regulating bone mineral density in middle-aged and older adults. However,whether there is a causal relationship between the cathepsin family and bone mineral density in other populations remains unknown. OBJECTIVE:To investigate the causal relationship between cathepsin and bone mineral density.METHODS:Genetic loci associated with eight cathepins were extracted from the IEU Open GWAS database as instrumental variables,and bone mineral density values in five age groups acted as an outcome. The causal relationship between cathepin and bone mineral density was assessed by two-way Mendelian randomization analysis. Heterogeneity of the genetic instrumental variables was assessed using Cochran's Q test,pleiotropy was assessed using the MR-Egger intercept test,and the sensitivity of single nucleotide polymorphisms used as instrumental variables to the causal effect of exposure and outcome was assessed using the leave-one-out method. RESULTS AND CONCLUSION:The results of the inverse variance weighting method with positive Mendelian randomization showed that cathepin H was negatively associated with bone mineral density in people aged 45-60 years[odds ratio (95％ confidence interval)=0.965(0.94-0.99),P=0.04];cathepin Z was negatively associated with bone mineral density in people aged 30-45 year[odds ratio (95％ confidence interval)=1.06 (1.00-1.11),P=0.03]. The results of sensitivity analysis showed a stable causal relationship,and MR-Egger intercept analysis did not detect potential horizontal pleiotropy. The inverse Mendelian randomization results showed that bone mineral density had no significant inverse effect on cathepin. The above results confirm that cathepin can affect bone mineral density in some age groups,which may increase the risk of osteoporosis and should be given more attention.

Objective The prediction of RR intervals in hypertensive patients can help clinicians to analyze and warn patients'heart condition.Methods Using 8 patients'data as samples,the RR intervals of patients were predicted by long short-term memory network(LSTM)and gradient lift tree(XGBoost),and the prediction results of the two models were combined by the inverse variance method to overcome the disadvantage of single model prediction.Results Compared with the single model,the proposed combined model had a different degree of improvement in the prediction of RR intervals in 8 patients.Conclusion LSTM-XGBoost model provides a method for predicting RR intervals in hypertensive patients,which has potential clinical feasibility.

Purpose/Significance To expound the content features of online medical reviews and its impact on the utilization of pa-tients'online consultation services,and to provide references for the information management of online medical platforms.Method/Process The LDA topic model based on TF-IDF is constructed,the semantic analysis method based on sentiment dictionary is adopted to mine the content features of online medical reviews,and the principal component analysis(PC A)is applied to classify and explore them,and the regression analysis is carried out by binary logistic.Result/Conclusion The content features of online medical reviews are divided into 3 categories,including 9 indicators.Among them,the attributes of reviewers,timeliness,consultation process,service atti-tude and platform feedback have a significant impact on the utilization of efficient online consultation services by patients.It is suggested to improve the utilization level of online consultation services by optimizing related services of online medical platform review section.