ObjectiveTo analyze the characteristics of 150 patients with spinal cord injury complicated with spasticity. MethodsA cross-sectional survey was conducted on 150 patients with spinal cord injury accompanied by spasticity from September, 2019 to December, 2024. Their age, gender, cause of injury, injury site, severity of injury, spasticity severity and other indicators were recorded. The relationships between different characteristics were analyzed, and a correlation analysis of disease duration, spasticity grade, injury level, injury severity and age were conducted. ResultsThere was no significant difference in age distribution between patients with tetraplegia and paraplegia (Z = 0.806, P = 0.420). The proportions of trauma (χ² = 3.982, P = 0.046) and tetraplegia (χ² = 10.559, P = 0.010) were higher in males than in females. Trauma was the main cause of injury in both tetraplegia and paraplegia patients; the proportion of tetraplegia was higher than paraplegia in trauma patients, while paraplegia was higher than tetraplegia in non-trauma patients (χ² = 11.885, P < 0.001). Patients with tetraplegia was dominated by incomplete injury, whereas patients with paraplegia was dominated by complete injury (χ² = 10.885, P = 0.012). Grade A injury was predominant in trauma patients (P = 0.003). Spasticity grade showed a very weak positive correlation with disease duration (r = 0.175, P = 0.032) and age (r = 0.168, P = 0.040). Injury severity showed a very weak positive correlation with age (r = 0.183, P = 0.025). ConclusionCharacteristics of patients with spinal cord injury complicated with spasticity is different with gender, cause of injury, injury level, injury severity.

BACKGROUND:The stemness index may be associated with the prognosis and immune infiltration of sarcoma,but the specific regulatory mechanism and characteristic genes have yet to be fully elucidated. OBJECTIVE:To investigate the correlation between stem cells and prognosis as well as immune infiltration in sarcoma employing the gene stemness index model and to identify the ferroptosis signature genes associated with sarcoma stem cells. METHODS:The sarcoma RNA sequencing data and related clinical information were obtained from the Cancer Genome Atlas(TCGA).The sarcoma RNA sequencing data were grouped using the sarcoma stemness index.Survival data were used to analyze prognosis between groups.Differentially expressed genes were obtained for pathway enrichment and immune infiltration analysis.Ferroptosis-related differential genes were used to construct a protein interaction network and analyze prognostic correlation.Rhabdomyosarcoma cell lines were cultured and divided into adherent cell group and stem cell group.The adherent cell group received no intervention,while the stem cell group was treated with serum-free culture to enrich stem cells in rhabdomyosarcoma cells.qRT-PCR was used to evaluate stemness markers,ferroptosis-related genes,and mRNA expression of ferroptosis-related markers in the cells. RESULTS AND CONCLUSION:(1)Patients were divided into high and low stemness index groups based on the median stemness index.The progression-free survival of patients in the high stemness index group was lower than that in the low stemness index group by disease risk prediction,suggesting poor prognosis.(2)According to GO and KEGG analysis,the groups with high and low stemness indices differed from one another.There were differences in immune infiltration between the high and low stemness index groups.Nine of the 23 ferroptosis-related genes in the differential genes have the potential to establish a highly correlated network of protein interactions.Patients with high expression of IDO1,IFNG,and AQP5 have a better prognosis,while those with high expression of CA9 have a poor prognosis.(3)The qRT-PCR results demonstrated a significant upregulation of stem cell-related markers NANOG,SOX2,and OCT4 mRNA expressions in the stem cell group compared to the adherent cell group(P<0.05).Compared to the adherent cell group,the stem cell group exhibited decreased mRNA expression level of ferroptosis-related marker SLC7A11(P<0.05)while showing increased levels of ACSL4,GPX4,FTH1,and COX2(P<0.05).Compared to the adherent cell group,the stem cell group displayed decreased mRNA expression level of differentially expressed gene CA9 alongside elevated levels of IDO1,IFNG,and AQP5(P<0.05).Stem cells were strongly associated with sarcoma survival and ferroptosis by bioinformatics analysis and experimental verification.Sarcoma stem cells have aberrant expression of CA9,IDO1,IFNG,and AQP5,which may serve as new targets for sarcoma therapy as well as diagnostic indicators.

Objective:To explore the effect and mechanism of naringenin (NAR) on glucose and lipid metabolism and oxidative stress in rats with gestational diabetes mellitus (GDM) .Methods:10 normal pregnant rats were selected as the control group, and the GDM model was prepared. One-time tail iv streptozotocin (STZ, 35 mg/kg) was used to construct GDM model. The 50 rats successfully modeled were divided into model group, metformin group (Met group, 20 mg/kg), low NAR (NAR-L, 50 mg/kg), high (NAR-H, 100 mg/kg) dose groups and NAR+EX527 group (NAR 100 mg/kg+EX527 10 mg/kg), 10 rats per group. Rats in Met group and NAR low-dose and high-dose groups were given corresponding doses of drugs ig. Rats in NAR+EX527 group were given NAR ig and EX527 ip at the same time, once a day, for 28 consecutive days. The levels of fasting blood glucose (FBG) and fasting serum insulin (FINS) in rats were detected, and the homeostasis model assessment for insulin resistance (HOMA-IR) was calculated; the levels of serum triglycerides (TG), cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were detected; the level of malondialdehyde (MDA) in pancreatic tissue and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected; the tissue lesions of pancreas and placenta were observed with HE staining; the expression of pancreatic tissue AMP-activated protein kinase (AMPK), p-AMPK, silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1 α) proteins was detected with Western blotting method. Results:Compared with control, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the model group were significantly increased, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly reduced ( P<0.05), and the placenta and pancreas tissues showed obvious pathological damage; Compared with the model group, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the Met group and the NAR-L and NAR-H groups were significantly reduced, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly increased ( P<0.05), the placenta and pancreas tissues damage reduced; and on the basis of NAR intervention, the use of SIRT1 inhibitor EX527 could significantly reverse the protective effect of NAR on GDM rats. Conclusion:NAR may improve glucose and lipid metabolism disorder and oxidative stress response in GDM rats by activatin SIRT1/PGC-1 α pathway.

Objective:To establish an in vivo infection model of West Nile virus (WNV) pseudovirus and evaluate the neutralizing activity of antibody WNV-XH1.Methods:A stable cell line that can package the WNV pseudovirus was established in the early stage to prepare the pseudovirus supernatant. The supernatant was concentrated and infected BHK21 cells to detect the titer of the pseudovirus. After intraperitoneal injection of the pseudovirus into C57BL/J mice, bioluminescence imaging was performed to observe the infection status of the pseudovirus in the mice. After simultaneous infection, blood was collected and ELISA was used to detect NS1 levels in mouse serum. The in vivo functional activity of antibody WNV-XH1 was evaluated using the established mouse infection model.Results:Fluorescence was detected in C57BL/J mice infected with WNV pseudovirus, and the NS1 levels in the peripheral blood serum of mice infected with pseudovirus were significantly higher than those of non infected mice (1.453±0.09vs0.305±0.018). After intravenous administration of WNV-XH1 antibody before the attack, the fluorescence signal in the mice decreased and the serum NS1 level decreased (0.384±0.015).Conclusions:A successful in vivo infection model of WNV pseudovirus was established, and it was confirmed that the antibody WNV-XH1 had a protective effect against WNV pseudovirus infection in vivo.

Objective:To establish an antibody expression system to reduce the antibody-dependent enhancement (ADE) effect of target antibody.Methods:Site-directed mutagenesis was used to mutate the 234 and 235 sites of the Fc region of the mammalian cell antibody expression vector-L234A and L235A to establish the antibody expression vector pFRT-IgG1κ-FcM. An antibody Wt-WNV with significant ADE effect obtained in previous work was selected and expressed by the pFRT-IgG1κ-FcM system to obtain mutant antibody FcM-WNV. The binding ability of FcM-WNV to target antigen West Nile virus envelope protein-DⅢ (WNV E-DⅢ) was detected by ELISA, and the its binding ability to human high-affinity IgG Fc receptor hFcγRⅠ (hCD64 ) was analyzed by flow cytometry. The neutralizing activity of FcM-WNV in vitro was detected by pseudovirus infection of host cells (BHK21 and K562). Results:The expression levels of FcM-WNV and Wt-WNV were comparable, and FcM-WNV could recognize and bind to WNVE-DIII in a concentration-dependent manner. Compared with Wt-WNV, the binding ability of FcM-WNV to hCD64 was significantly weakened, showing a significant decrease in fluorescence intensity. Consistent with the previous experimental results, Wt-WNV at a concentration of 5 μg/ml significantly enhanced the infection of K562 by WNV pseudovirus, while FcM-WNV at a concentration of 5 μg/ml could effectively block pseudovirus infection in both K562 and BHK21 cells.Conclusions:The established antibody expression system can effectively reduce the ADE effect of the target antibody.

Objective To investigate the molecular mechanism of ferroptosis in glioblastoma(GBM)and to provide insights for identifying new therapeutic targets.Methods GSE108474 was selected from gene expression omnibus(GEO)database and differentially expressed genes(DEGs)in GBM were obtained by using GEO2R,compared with the gene set in the Ferroptosis database(FerrDb)to identify ferroptosis-related gene.GO and KEGG enrich-ment analyses were conducted using DAVID database.A protein-protein interaction network was created using String website.Hub genes with high connectivity were confirmed using Cytoscape software.Prognostic and immune infiltration analyses were performed using TIMER website.RNA expression levels and gene correlation analyses were carried out using GEPIA website.Differential expression of hub gene proteins was analyzed by using the HPA database.Tumor immune characteristic correlations were examined using TISIDB database.Differences in mRNA expression of hub genes between tumor cells A172 and U251MG and normal astrocytes HA1800 were compared u-sing the quantitative real-time PCR.Results Out of 5 331 differentially expressed genes,114 were related to fer-roptosis.GO and KEGG enrichment analysis suggested that these 114 genes might play roles in positive regulation of gene expression,and affect tumor progression through ferroptosis and autophagy pathways.10 hub genes were i-dentified in the protein-protein interaction network,among which cluster of differentiation 44(CD44),murine double minute 2(MDM2)and signal transducer and activator of transcription 3(STAT3)were found to be highly expressed in tumors with lower survival rates.CD44,MDM2 and STAT3 mRNA expression were higher in GBM cells compared to normal cells.Protein expression of CD44,MDM2 and STAT3 was higher in high-grade glioma tis-sues than that in normal tissues.The expression of three genes in the tumor was negatively correlated with ferropto-sis.Immune infiltration analysis revealed that CD44,MDM2 and STAT3 in the tumor were related to the infiltration of neutrophils,CD4+T cells,and dendritic cells,and the expression of three genes was related to various chemo-kines and their receptors.Conclusion CD44,MDM2 and STAT3 may play a role in tumor ferroptosis and immune regulation,which have the potential to become a therapeutic target for GBM.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To explore the effect and mechanism of naringenin (NAR) on glucose and lipid metabolism and oxidative stress in rats with gestational diabetes mellitus (GDM) .Methods:10 normal pregnant rats were selected as the control group, and the GDM model was prepared. One-time tail iv streptozotocin (STZ, 35 mg/kg) was used to construct GDM model. The 50 rats successfully modeled were divided into model group, metformin group (Met group, 20 mg/kg), low NAR (NAR-L, 50 mg/kg), high (NAR-H, 100 mg/kg) dose groups and NAR+EX527 group (NAR 100 mg/kg+EX527 10 mg/kg), 10 rats per group. Rats in Met group and NAR low-dose and high-dose groups were given corresponding doses of drugs ig. Rats in NAR+EX527 group were given NAR ig and EX527 ip at the same time, once a day, for 28 consecutive days. The levels of fasting blood glucose (FBG) and fasting serum insulin (FINS) in rats were detected, and the homeostasis model assessment for insulin resistance (HOMA-IR) was calculated; the levels of serum triglycerides (TG), cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were detected; the level of malondialdehyde (MDA) in pancreatic tissue and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected; the tissue lesions of pancreas and placenta were observed with HE staining; the expression of pancreatic tissue AMP-activated protein kinase (AMPK), p-AMPK, silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1 α) proteins was detected with Western blotting method. Results:Compared with control, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the model group were significantly increased, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly reduced ( P<0.05), and the placenta and pancreas tissues showed obvious pathological damage; Compared with the model group, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the Met group and the NAR-L and NAR-H groups were significantly reduced, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly increased ( P<0.05), the placenta and pancreas tissues damage reduced; and on the basis of NAR intervention, the use of SIRT1 inhibitor EX527 could significantly reverse the protective effect of NAR on GDM rats. Conclusion:NAR may improve glucose and lipid metabolism disorder and oxidative stress response in GDM rats by activatin SIRT1/PGC-1 α pathway.

Objective:To establish an in vivo infection model of H5N1 pseudovirus and to detect the neutralizing activity of FHA3 antibody using this model. Methods:Based on the sequence information of hemagglutinin (HA) and neuraminidase (NA) of A/Anhui/1/2005/H5N1 strain, two recombinant plasmids of pcDNA3.1-HA5 and pcDNA3.1-NA1 were constructed. The two plasmids and plasmid pNL4-3.Luc.R-E- were co-transfected into 293T cells to prepare H5N1 pseudovirus supernatant. The morphology of pseudovirus particles in the supernatant was observed by electron microscopy. MDCK cells were infected with the pseudovirus supernatant and the virus titer was detected. BALB/c mice were injected with the pseudovirus supernatant by intraperitoneal injection and subjected to bioluminescence imaging at 2, 5, 8, and 12 d after infection to detect the pseudovirus infection in vivo. The functional activity of FHA3 antibody in vivo was evaluated using the established mouse infection model. Results:The recombinant plasmids pcDNA3.1-HA5 and pcDNA3.1-NA1 were correctly constructed and could be used to prepare pseudovirus supernatants of high titer by co-transfecting 293T cells with the plasmid pNL4-3.Luc.R-E-. The virus particles were round under electron microscope. H5N1 pseudovirus-infected mice exhibits strong fluorescence signals, which were attenuated by FHA3 treatment before challenge.Conclusions:The in vivo infection model of H5N1 pseudovirus was successfully constructed and FHA3 antibody was proved to be protective against the pseudovirus infection.

ObjectiveTo observe the characteristics of gait symmetry and its influencing factors in patients with incomplete spinal cord injury (ISCI). MethodsFrom May, 2018 to November, 2021, 34 patients with ISCI in Beijing Bo'ai Hospital were divided into symmetrical injury of lower limb (SI) group and asymmetrical injury of lower limb (ASI) group according to the lower extremities motor score (LEMS). Three dimensional motion acquisition system and plantar pressure acquisition system were used for gait test. The symmetry indexes of step length, stance time and swing time were caculated. ResultsThe symmetry indexes of step length, stance time and swing time were significant lower in SI group than in ASI group (|t| > 2.619, P < 0.01). Stance time and swing time significantly correlated to the difference of bilateral LEMS in ASI group (r > 0.468, P < 0.01). Discriminant analysis showed that gait parameter equations were different for patients with different symmetry of lower limb injuries. ConclusionThe symmetry of lower limb motor function impacts gait symmetry for patients with ISCI, especially the difference value of bilateral total LEMS. Gait parameters can be used to determine the symmetry of lower limb injury in patients with ISCI.