OBJECTIVE: To prepare decellularized nerve grafts from alpha-1, 3-galactosyltransferase (GGTA1) gene-edited pigs and explore their biocompatibility for xenotransplantation. METHODS: The sciatic nerves from wild-type pigs and GGTA1 gene-edited pigs were obtained and underwent decellularization. The alpha-galactosidase (α-gal) content in the sciatic nerves of GGTA1 gene-edited pigs was detected by using IB4 fluorescence staining and ELISA method to verify the knockout status of the GGTA1 gene, and using human sciatic nerve as a control. HE staining and scanning electron microscopy observation were used to observe the structure of the nerve samples. Immunofluorescence staining and DNA content determination were used to evaluate the degree of decellularization of the nerve samples. Fourteen nude mice were taken, and subcutaneous capsules were prepared on both sides of the spine. Decellularized nerve samples of wild-type pigs ( n=7) and GGTA1 gene-edited pigs ( n=7) were randomly implanted in the subcutaneous capsules. Blood was drawn at 1, 3, 5, and 7 days after implantation to detect neutrophil counting. RESULTS: IB4 fluorescence staining and ELISA detection showed that GGTA1 gene was successfully knocked out in the nerves of GGTA1 gene-edited pigs. HE staining showed that the structure of the decellularized nerve from GGTA1 gene-edited pigs was well preserved; the nerve basement membrane tube structure was visible under scanning electron microscopy; no cell nuclei was observed, and the extracellular matrix components was retained in the nerve grafts by immunofluorescence staining; and the DNA content was significantly reduced when compared with the normal nerves ( P<0.05). In vivo experiments showed that the number of neutrophils in the two groups were similar at 1, 3, and 7 days after implantation, with no significant difference ( P>0.05); only at 5 days, the number of neutrophils was significantly lower in the GGTA1 gene-edited pigs than in the wild-type pigs ( P<0.05). CONCLUSION The decellularized nerve grafts from GGTA1 gene-edited pigs have well-preserved nerve structure, complete decellularization, retain the natural nerve basement membrane tube structure and components, and low immune response after xenotransplantation through in vitro experiments.
Animals ; Transplantation, Heterologous ; Galactosyltransferases/genetics* ; Sciatic Nerve/immunology* ; Swine ; Tissue Engineering/methods* ; Humans ; Graft Rejection/prevention & control* ; Gene Editing ; Mice ; Mice, Nude ; Heterografts/immunology* ; Animals, Genetically Modified ; Tissue Scaffolds ; Decellularized Extracellular Matrix

BACKGROUND:Polylactic acid(PLA)has good biocompatibility and a controllable degradation rate and is currently widely used in biomedical engineering.However,PLA has shortcomings such as low mechanical strength and insufficient biological activity,which limits its further application in bone tissue engineering. OBJECTIVE:To construct polylactic acid/polydopamine/tantalum(PLA/PDA/Ta)bone tissue engineering scaffolds,and explore their biosafety and in vitro osteogenesis. METHODS:A PLA scaffold with a porous structure was prepared through liquid crystal display light-curing technology.PLA/PDA scaffolds and PLA/PDA/Ta scaffolds were prepared by soaking PLA scaffolds in dopamine solution and dopamine-tantalum nanoparticle solution,respectively.The microstructure and water contact angle of scaffolds were characterized.MC3T3-E1 cells were co-cultured with PLA,PLA/PDA,and PLA/PDA/Ta scaffolds,respectively,and CCK-8 assay and live/dead cell staining were performed.After osteogenic differentiation,alkaline phosphatase,alizarin red staining,and osteogenic gene detection were performed. RESULTS AND CONCLUSION:(1)The scanning electron microscope results exhibited that the three kinds of prepared scaffolds had an interconnected porous three-dimensional structure,and the average pore diameter was 200 μm.The water contact angle of PLA/PDA/Ta scaffolds was lower than that of PLA and PLA/PDA scaffolds(P<0.05).(2)CCK-8 assay showed that compared with PLA and PLA/PDA scaffolds,PLA/PDA/Ta scaffolds could promote cell proliferation(P<0.05).Live/dead cell staining showed good cell proliferation in the three groups.(3)Alkaline phosphatase and alizarin red staining showed that compared with PLA and PLA/PDA scaffolds,PLA/PDA/Ta scaffolds could promote the expression of alkaline phosphatase and the formation of mineralized nodules.RT-qPCR showed that compared with PLA and PLA/PDA scaffolds,PLA/PDA/Ta scaffolds could enhance the mRNA expression of cell bone morphogenetic protein,Runx-2,and type I collagen(P<0.05,P<0.01).(4)The results showed that the PLA/PDA/Ta scaffold had excellent osteogenic activity and the ability to promote cell proliferation.

Objective To investigate the effects of early pulmonary rehabilitation training on lung function recovery,complication rates,and quality of life in patients who undergo thoracoscopic surgery.Methods A ran-domized controlled trial was conducted involving 132 patients who underwent thoracoscopic surgery between June 2020 and June 2023.Participants were randomly allocated to either an early pulmonary rehabilitation program(n=72)or a conventional rehabilitation program(n=60).The early intervention group commenced a staged rehabilitation protocol—including respiratory training and physical exercise—within 24 to 72 hours following surgery,whereas the conventional group received standard postoperative nursing care only.Outcomes measured included postoperative pulmonary function(FEV1,FVC),incidence of postoperative complications,length of hospi-tal stay,and quality of life scores(maximum score:40).These outcomes were compared between the two groups.Results On postoperative days 3 and 7,the early group showed significantly higher FEV1 values compared to the conventional group[(1.6±0.17)L vs.(1.3±0.21)L;(1.9±0.20)L vs.(1.6±0.11)L,respectively],as well as improved FVC measurements[(2.5±0.20)L vs.(2.1±0.14)L;(2.9±0.25)L vs.(2.3±0.23)L(all P＜0.05)].The early intervention group also demonstrated a significantly lower overall complication rate(14.3％vs.33.3％,x2=6.79,P=0.009),including reduced incidences of atelectasis(6.9％vs.20.0％,P=0.031)and pulmonary infection(4.2％vs.13.3％,P=0.044).Additionally,patients in the early group had a shorter average hospital stay[(8.4±1.2)days vs.(10.9±2.3)days,P=0.018]and reported higher quality of life scores[(35.6±3.7)vs.(30.8±4.5),P=0.032].No significant difference was observed between the two groups in terms of the overall incidence of adverse events(23.6％vs.31.7％,x2=1.07,P=0.301).Conclusions Early pulmonary rehabilitation significantly facilitates the recovery of lung function,reduces the incidence of postoperative complica-tions,enhances quality of life,and demonstrates a favorable safety profile among patients undergoing thoracoscopic surgery.Therefore,this intervention should be more broadly implemented in clinical practice to optimize postop-erative recovery outcomes.

Objective:To synthesize evidence regarding discharge preparation services for patients with severe acute pancreatitis (SAP) .Methods:Systematic search of relevant domestic and international websites and databases for diverse evidence concerning discharge preparation in SAP patients were conducted. Literature quality of the included studies was evaluated, and related evidence was extracted and graded.Results:Based on the researchers' evaluation and synthesis, 25 pieces of evidence related to discharge preparation services for patients with SAP were identified and summarized. These were categorized into five key areas: comprehensive assessment, care planning, multidimensional interventions, post-discharge follow-up, and ongoing evaluation. Among them, the evidence with stronger recommendation levels included multidisciplinary team assessment within 24 hours of admission, the assignment of dedicated personnel to coordinate, monitoring the discharge plan and continuous evaluation of the effectiveness of discharge preparation services.Conclusions:This study summarizes the best evidence available regarding discharge preparation services for SAP patients. It may provide an evidence-based foundation for nursing staff to standardize clinical practice and to further develop discharge preparation programs.

Objective To study the development of conus medullaris by quantitative three-dimensional ultrasound in the second and third trimesters of pregnancy,and to analyze the value of terminal caudal vertebral ossification center count in diagnosis of closed spina bifida.Methods A total of 108 normal fetuses in the second and third trimesters of pregnancy who were admitted to Baoding No.1 Central Hospital from January 2019 to March 2024 were selected and assigned to the normal group.The distance between the fetal conus medullaris and the first sacral vertebra,the distance between the end of the conus medullaris and the end of sacrum,and the number of corresponding vertebrae were measured by three-dimensional ultrasound at different gestational weeks.Meanwhile,54 fetuses who were confirmed to have closed spina bifida after delivery were enrolled into the abnormal group.The number of ossification centers on the median sagittal section of the caudal spine was counted and compared between the two groups.The receiver operating characteristic(ROC)curve was drawn to analyze the value of ossification center count of distal caudal vertebrae in diagnosis of closed spina bifida.Results With the increase of gestational weeks,the distance between the fetal conus medullaris and the first sacral vertebra and the distance between the end of the conus medullaris and the end of the sacral bone were gradually increased.The conus medullaris of normal fetuses was mainly located in L1.5,L2 and L2.5,accounting for 85.19%(92/108).The ossification center count of distal caudal vertebra was slightly increased with the increase of gestational weeks in the normal group.At 17-20 gestational weeks,85.71%(12/14)of the ossification center count of distal caudal vertebra was 5 to 7.At 21-28 gestational weeks,100.00%(45/45)of the ossification center count was 6 and more.At week 29 and above,100.00%(45/45)of the ossification center count was more than 7.In the abnormal group,96.30%(52/54)of the fetuses had the ossification center less than 6,and 85.71%(42/49)of the ossification center count was≤5 at 21-32 gestational weeks.There were significant differences in the ossification center count between the two groups at 17-20,21-24,25-28,29-32,and 33-36 gestational weeks(P<0.05).ROC curve showed that,at 17-20,21-24,25-28,29-32,and 33-36 gestational weeks,the area under the curve(AUC)in prediction of closed spina bifida was 0.804,0.744,0.776,0.819,and 0.722 when the ossification center count of distal caudal vertebra was 5.31,4.59,4.81,4.67,and 5.49 as the cut-off value,respectively.Conclusion The conus medullaris moves upward with the increase of gestational weeks in the second and third trimesters of pregnancy.The ossification center count of the distal caudal vertebra is less than 6 in most fetuses with closed spina bifida,which is conducive to screening for closed spina bifida.

Objective To investigate the effects of early pulmonary rehabilitation training on lung function recovery,complication rates,and quality of life in patients who undergo thoracoscopic surgery.Methods A ran-domized controlled trial was conducted involving 132 patients who underwent thoracoscopic surgery between June 2020 and June 2023.Participants were randomly allocated to either an early pulmonary rehabilitation program(n=72)or a conventional rehabilitation program(n=60).The early intervention group commenced a staged rehabilitation protocol—including respiratory training and physical exercise—within 24 to 72 hours following surgery,whereas the conventional group received standard postoperative nursing care only.Outcomes measured included postoperative pulmonary function(FEV1,FVC),incidence of postoperative complications,length of hospi-tal stay,and quality of life scores(maximum score:40).These outcomes were compared between the two groups.Results On postoperative days 3 and 7,the early group showed significantly higher FEV1 values compared to the conventional group[(1.6±0.17)L vs.(1.3±0.21)L;(1.9±0.20)L vs.(1.6±0.11)L,respectively],as well as improved FVC measurements[(2.5±0.20)L vs.(2.1±0.14)L;(2.9±0.25)L vs.(2.3±0.23)L(all P＜0.05)].The early intervention group also demonstrated a significantly lower overall complication rate(14.3％vs.33.3％,x2=6.79,P=0.009),including reduced incidences of atelectasis(6.9％vs.20.0％,P=0.031)and pulmonary infection(4.2％vs.13.3％,P=0.044).Additionally,patients in the early group had a shorter average hospital stay[(8.4±1.2)days vs.(10.9±2.3)days,P=0.018]and reported higher quality of life scores[(35.6±3.7)vs.(30.8±4.5),P=0.032].No significant difference was observed between the two groups in terms of the overall incidence of adverse events(23.6％vs.31.7％,x2=1.07,P=0.301).Conclusions Early pulmonary rehabilitation significantly facilitates the recovery of lung function,reduces the incidence of postoperative complica-tions,enhances quality of life,and demonstrates a favorable safety profile among patients undergoing thoracoscopic surgery.Therefore,this intervention should be more broadly implemented in clinical practice to optimize postop-erative recovery outcomes.

Objective:To develop an original-mirror alignment associated deep learning algorithm for intelligent registration of three-dimensional maxillofacial point cloud data,by utilizing a dynamic graph-based registration network model(maxillofacial dynamic graph registration network,MDGR-Net),and to provide a valuable reference for digital design and analysis in clinical dental applications.Methods:Four hundred clinical patients without significant deformities were recruited from Peking University School of Stomatology from October 2018 to October 2022.Through data augmentation,a total of 2 000 three-dimensional maxillofacial datasets were generated for training and testing the MDGR-Net algorithm.These were divided into a training set(1 400 cases),a validation set(200 cases),and an internal test set(200 cases).The MDGR-Net model constructed feature vectors for key points in both original and mirror point clouds(X,Y),established correspondences between key points in the X and Y point clouds based on these feature vectors,and calculated rotation and translation matrices using singular value decomposi-tion(SVD).Utilizing the MDGR-Net model,intelligent registration of the original and mirror point clouds were achieved,resulting in a combined point cloud.The principal component analysis(PCA)algorithm was applied to this combined point cloud to obtain the symmetry reference plane associated with the MDGR-Net methodology.Model evaluation for the translation and rotation matrices on the test set was performed using the coefficient of determination(R2).Angle error evaluations for the three-dimensional maxillofacial symmetry reference planes were constructed using the MDGR-Net-associated method and the"ground truth"iterative closest point(ICP)-associated method were conducted on 200 cases in the inter-nal test set and 40 cases in an external test set.Results:Based on testing with the three-dimensional maxillofacial data from the 200-case internal test set,the MDGR-Net model achieved an R2 value of 0.91 for the rotation matrix and 0.98 for the translation matrix.The average angle error on the internal and external test sets were 0.84°±0.55° and 0.58°±0.43°,respectively.The construction of the three-dimensional maxillofacial symmetry reference plane for 40 clinical cases took only 3 seconds,with the model performing optimally in the patients with skeletal Class Ⅲ malocclusion,high angle cases,and Angle Class Ⅲ orthodontic patients.Conclusion:This study proposed the MDGR-Net association method based on intelligent point cloud registration as a novel solution for constructing three-dimensional maxillo-facial symmetry reference planes in clinical dental applications,which can significantly enhance diagnos-tic and therapeutic efficiency and outcomes,while reduce expert dependence.

OBJECTIVE To improve the efficiency and quality of dispensed oral drug management in the inpatient pharmacy,and ensure the safety of drug use in patients.METHODS SWOT(strength,weakness,opportunity,threat)analysis method was used to analyze the internal strengths and weaknesses,as well as the external opportunities and threats in the construction of a closed-loop management system for dispensed oral drugs in the inpatient pharmacy of our hospital,and propose improvement strategies.RESULTS&CONCLUSIONS A refined,full-process,closed-loop traceability management system for dispensed oral drugs in the inpatient pharmacies was successfully established,which is traceable in origin,trackable in destination,and accountable in responsibility.After the application of this system,the registration rate of dispensed drug information and the correctness rate of registration content both reached 100%.The proportion of overdue drug varieties in the same period of 2024 decreased by 77.78%compared to March 2020,the inventory volume decreased by 29.50%compared to the first quarter of 2020,the per-bed medication volume decreased by 32.14%compared to the first quarter of 2020;the average workload per post in the same period of 2023 increased by 49.09%compared to 2019,the dispensing accuracy rate reached 100%,and the improvement rate of quality control problem increased by 25.25%compared to 2021.This system effectively improves the safety and accuracy of dispensed oral drug management in the inpatient pharmacy.

BACKGROUND:Studies have shown that immune cells are involved in all processes of ischemic stroke,in which cuproptosis also plays a key role.OBJECTIVE:To screen diagnostic biomarkers related to the progression of ischemic stroke through bioinformatics,and analyze and validate cuproptosis-related genes closely related to the occurrence and development of ischemic stroke.METHODS:The GSE16561 microarray was obtained from the GEO database,containing data from 39 cases of ischemic stroke(ischemic stroke group)and 24 controls(control group).Differentially expressed genes from the ischemic stroke microarray data were analyzed.Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed.By using LASSO and Random Forest methods,key genes affecting the occurrence and development of ischemic stroke were screened,and a diagnostic model was established and validated.Differential gene analysis was performed through immune cell infiltration and weighted gene co expression network.The differentially expressed immune-related genes were intersected with cuproptosis genes to obtain the hub genes related to cuproptosis immunity.In vitro cell experiments were conducted to divide rat hippocampal neurons into a normal group and an ischemic stroke group,and qPCR experiments were performed to verify the results.RESULTS AND CONCLUSION:(1)573 differentially expressed genes were obtained by differential analysis.Differentially expressed genes were mainly enriched in biological processes,such as positive regulation of immune response,and signaling pathways such as lipid and atherosclerosis.(2)Machine learning methods were used to screen diagnostic genes such as MFN2,PKM2,CREG1,and FOXO3A,which may have some diagnostic value for ischemic stroke.(3)Immune infiltration analysis revealed resting plasma cells,NK cells,macrophages,etc.,indicating that immune cells play a certain role in the pathogenesis of ischemic stroke.(4)Weighted gene co-expression network analysis combined with immune infiltration analysis obtained 118 key module genes,which were intersected with cuproptosis genes to obtain 2 cuproptosis and immune characteristic genes.The correlation analysis between four diagnostic genes and Hub genes showed that the expression of FOXO3A and MFN2,PKM2 and BCL2L1,MTF1 and MFN2,ATP7B and BCL2L1 were correlated.(5)The qPCR results showed significant differences in the genes MTF1 and ATP7B between the ischemic stroke group and the blank group.To conclude,ATP7B and MTF1 can serve as characteristic genes for cuproptosis in ischemic stroke.It is possible to improve ischemic stroke by intervening in ATP7B and MTF1 to regulate cuproptosis.

Objective:To synthesize evidence regarding discharge preparation services for patients with severe acute pancreatitis (SAP) .Methods:Systematic search of relevant domestic and international websites and databases for diverse evidence concerning discharge preparation in SAP patients were conducted. Literature quality of the included studies was evaluated, and related evidence was extracted and graded.Results:Based on the researchers' evaluation and synthesis, 25 pieces of evidence related to discharge preparation services for patients with SAP were identified and summarized. These were categorized into five key areas: comprehensive assessment, care planning, multidimensional interventions, post-discharge follow-up, and ongoing evaluation. Among them, the evidence with stronger recommendation levels included multidisciplinary team assessment within 24 hours of admission, the assignment of dedicated personnel to coordinate, monitoring the discharge plan and continuous evaluation of the effectiveness of discharge preparation services.Conclusions:This study summarizes the best evidence available regarding discharge preparation services for SAP patients. It may provide an evidence-based foundation for nursing staff to standardize clinical practice and to further develop discharge preparation programs.