Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

An analysis of 24 144 norovirus sequences from Asia and Russia deposited in GenBank between 1995 and 2023 was conducted,to understand the temporal and spatial variations in norovirus genotypes in these regions.Norovirus sequences from Asia and Russia were downloaded in FASTA format from GenBank for the years 1995-2023,and analyzed in Excel,R language,and GraphPad Prism for data visualization.The number of norovirus sequences submitted to GenBank increased annually from 2004 and peaked in 2015.Notably,China and Japan contributed 62.3％of all submitted norovirus sequences.These sequences encompassed 31 capsid genotypes(C-type),with GⅠ accounting for 9％and GⅡ accounting for 90％.Additionally,49 polymerase types(P-type)were identified,along with 68 combinations of CP types;among the analyzed recombinant sequences(4 460 entries in total),approxi-mately 41％belonged to three predominant recombinant strains:GⅡ.2[P16],GⅡ.4[P31],and GⅡ.4[P16].This analysis provides valuable insights into the distribution characteristics of norovirus genotypes across Asia and Russia over time,thereby supporting vac-cine design and evaluation efforts.

Objective To explore the protective effects of genetically modified porcine erythrocyte suspension as a subnormothermic normoxic mechanical perfusate on hypoxic-ischemic brain injury in cynomolgus monkeys caused by traumatic hemorrhage.Methods Cynomolgus monkeys were randomly divided into positive and negative control groups(a total of 3 monkeys,with 3 left cerebral hemispheres as the positive control group and 3 right cerebral hemispheres as the negative control group)and the subnormothermic perfusion group(n=3).The positive control group was directly sampled 1 hour after circulatory arrest,while the negative control group was placed at subnormothermic conditions for 6 hours after circulatory arrest.The subnormothermic perfusion group underwent 6 hours of subnormothermic normoxic mechanical perfusion of the bilateral common carotid arteries of the cynomolgus monkey hypoxic-ischemic brain injury model using genetically modified porcine erythrocyte suspension 1 hour after circulatory arrest.Before perfusion,cross-matching experiments were conducted between the six genetically modified pig and the cynomolgus monkeys.After the start of perfusion,the levels of routine blood indicators in the perfusate were detected at 0,1,2,3,4,5 and 6 hours.Blood oxygen saturation was recorded,and the levels of Na+,K+,Ca2+,glucose and blood pH in the perfusate were measured,as well as the levels of IgG and IgM in the perfusate.After 6 hours of perfusion,the water content of the brain tissue was measured.Nissl staining was performed on the frontal cortex and hippocampal regions,and immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP),ionized calcium-binding adapter molecule 1(Iba1)and neuronal nuclear antigen(NEUN).Results The cross-matching results between the six genetically modified pig and the cynomolgus monkeys were negative.The number of red blood cells in the perfusate decreased significantly at 3 hours of perfusion,and the hemoglobin level showed a downward trend at 1,3,5 and 6 hours.The number of white blood cells and platelets decreased at all time points.The blood oxygen saturation in the subnormothermic perfusion group remained stable at 95%-98%,and the levels of blood oxygen saturation,Na+,Ca2+,glucose and pH were stable,while the K+level first increased and then decreased.There was no significant difference in the levels of IgG and IgM before and after perfusion.The water content of brain tissue at the end of perfusion in the subnormothermic perfusion group was significantly higher than that in the positive control group(P<0.001).Nissl staining results showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex of the subnormothermic perfusion group maintained better morphological integrity,with no significant increase in enlarged and deformed cells.In the hippocampal CA1 region,there was a slight increase in enlarged and deformed cells,and a few cells with undamaged structures showed reduced cell size.In the hippocampal dentate gyrus,fewer granule neurons had compromised structural integrity,with increased cell edema.NEUN immunofluorescence staining showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex and hippocampal CA1 region of the subnormothermic perfusion group had better morphological states,with clear axons.The granule cells in the hippocampal dentate gyrus were well preserved,but the nuclei were less well protected.GFAP immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had sparser protrusions that were more tightly associated with neurons.Iba1 immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had thicker and fewer protrusions.Conclusions Compared with the positive control group,subnormothermic normoxic mechanical perfusion with genetically modified porcine erythrocyte perfusate increases brain tissue edema in cynomolgus monkeys,but better preserves the morphological integrity of neurons and glial cells.The protective effects may be related to the continuous oxygen and energy supply,maintenance of ion homeostasis and perfusate pH,reduced rejection,and low metabolic state of the whole brain.

Objective To investigate a cluster of healthcare-associated infection(HAI)of carbapenem-resistant Klebsiella pneumoniae(CRKP)in intensive care unit(ICU)of a traditional Chinese medicine hospital,and provide evidence for the prevention and control of multidrug-resistant organisms in hospitals.Methods From January to Ju-ly 2023,multiple cases of CRKP infection in the ICU of a traditional Chinese medicine hospital occurred,surpassing the number in the same period of previous years.Through collection of patients'clinical information,epidemiologi-cal investigation and environmental hygiene monitoring,the isolated CRKP strains were identified by pulsed-field gel electrophoresis(PFGE),possible causes of transmission were found out,and corresponding control measures were taken.Results During the study period,a total of 13 patients were confirmed to have developed CRKP infection,mainly respiratory tract infection,bloodstream infection,and abdominal infection.Environmental monitoring showed that only 2 out of 131 environmental object surfaces specimens and staff's hand specimens tested positive for CRKP.PFGE identification showed that the 2 CRKP strains isolated from the sink specimens were highly homolo-gous to the strains isolated from 2 recently admitted patients.Some strains isolated from patients were highly ho-mologous,indicating a possibility of cross transmission.Conclusion CRKP is prone to be cross-transmission among ICU patients and contaminates the environment.Early warning and investigation should be conducted to prevent the occurrence of HAI outbreaks.

Objective To utilize the dental implant robot in the clinical instruction of oral implantology in order to popularize the technology and serve the medical education.Methods The effectiveness of the robotics teaching approach was assessed through a questionnaire.Results The dental implant robot technology has certain advantages in the clinical teaching of oral implant majors.Conclusion With the increasing popularity of digital implantation,the emerging dental implant robot has a wide range of applications and many advantages.It is very necessary to introduce it into the clinical teaching of oral implant majors.

Objective To explore the optimisation path of medical resource allocation efficiency improvement in Traditional Chinese Medicine (TCM) hospitals under the synergistic effect of multiple factors,so as to provide powerful support for the balanced development of medical resources in TCM hospitals.Methods The comprehensive efficiency of resource allocation in TCM hospitals in 31 provinces (cities and districts) of China obtained by Data Envelopment Analysis was taken as the outcome variable,and with the help of Fuzzy-set Qualitative Comparative Analysis,it examined the configurations of efficient medical resource allocation,considering the following conditional variables:per capital Regional GDP,the proportion of fiscal allocation revenue to total income,the ratio of TCM practicing (assistant) physicians to all practicing (assistant) physicians,the average length of hospital stay for discharged patients,the number of total diagnosis and treatments per thousand population,and the number of hospital beds per thousand population.Results By analyzing the conditional patterns of efficient allocation of medical resources in TCM hospitals,three equivalent driving paths can be summarized,which are comprehensive service capacity,TCM advantage and hospitalization driving paths.Conclusion The overall level of medical resource allocation efficiency of TCM hospitals in China needs to be improved.In the future,efforts should be made to improve the comprehensive service capacity and operational efficiency of hospitals,give full play to the advantages of TCM,build a high-quality TCM talent team,reasonably shorten the average hospital stay,and improve the utilization efficiency of hospital beds.

Objective To explore the effects of ultra-low tube voltage of the third-generation dual-source CT combined with advanced modeled iterative reconstruction(ADMIRE)in bronchial artery CTA for hemoptysis patients.Methods Totally 70 hemoptysis patients admitted to some hospital from March to August 2023 had their clinical and imaging data collected retrospectively,and then divided into Group A and B with the numerical table method.In Group A image reconstruction was carried out with 120 kV tube voltage and filtered back projection(FBP)(named as Group A-FBP);In Group B image reconstruction was performed with 70 kV tube voltage respectively combined with FBP,ADMIRE Ⅲ,Ⅳ and Ⅴ(named as Group B-FBP,B-ADMIRE Ⅲ,B-ADMIRE Ⅳ and B-ADMIRE Ⅴ).All the patients were scanned with Siemens third-generation dual-source CT.The groups were compared in terms of radiation dose covering volume CT dose index,dose length product and effective dose,subjective score and objective evaluation.The subjective score agreement between 2 radiologists was evaluated with Kappa test.SPSS 26.0 software was used for statistical analysis.Results The radiation dose parameters of Group B were all lower than those of Group A,and the differences were statistically significant(P<0.001).Group B-FBP and ADMIRE Ⅳ had the lowest and highest subjective scores for bronchial artery images,res-pectively,and the five groups were significantly different in subjective score(P<0.05).The two radiologists had high agree-ment for Kappa values of the subjective scores,with the Kappa values being not lower than 0.75.Group A-FBP showed the lowest vessel CT value,Group B-FBP exhibited the highest image noise and the lowest SNR and CNR,and the five groups differed significantly in the objective evaluation of image quality(P<0.05).Group B-ADMIRE Ⅲ,B-ADMIRE Ⅳ and B-ADMIRE Ⅴ had statistically significant differences with Group A-FBP in objective evaluation of image quality(all P<0.05),with Group B-ADMIRE Ⅴ exhibiting the lowest image noise while highest SNR and CNR.Conclusion Ultra-low tube voltage of the third-generation dual-source CT combined with ADMIRE decreases the radiation dose and ensures image quality when applied to hemoptysis patient CTA,and ADMIRE Ⅳ behaves well in noise reduction and image quality enhancement.[Chinese Medical Equipment Journal,2025,46(10):60-64]

Objective To explore the protective effects of genetically modified porcine erythrocyte suspension as a subnormothermic normoxic mechanical perfusate on hypoxic-ischemic brain injury in cynomolgus monkeys caused by traumatic hemorrhage.Methods Cynomolgus monkeys were randomly divided into positive and negative control groups(a total of 3 monkeys,with 3 left cerebral hemispheres as the positive control group and 3 right cerebral hemispheres as the negative control group)and the subnormothermic perfusion group(n=3).The positive control group was directly sampled 1 hour after circulatory arrest,while the negative control group was placed at subnormothermic conditions for 6 hours after circulatory arrest.The subnormothermic perfusion group underwent 6 hours of subnormothermic normoxic mechanical perfusion of the bilateral common carotid arteries of the cynomolgus monkey hypoxic-ischemic brain injury model using genetically modified porcine erythrocyte suspension 1 hour after circulatory arrest.Before perfusion,cross-matching experiments were conducted between the six genetically modified pig and the cynomolgus monkeys.After the start of perfusion,the levels of routine blood indicators in the perfusate were detected at 0,1,2,3,4,5 and 6 hours.Blood oxygen saturation was recorded,and the levels of Na+,K+,Ca2+,glucose and blood pH in the perfusate were measured,as well as the levels of IgG and IgM in the perfusate.After 6 hours of perfusion,the water content of the brain tissue was measured.Nissl staining was performed on the frontal cortex and hippocampal regions,and immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP),ionized calcium-binding adapter molecule 1(Iba1)and neuronal nuclear antigen(NEUN).Results The cross-matching results between the six genetically modified pig and the cynomolgus monkeys were negative.The number of red blood cells in the perfusate decreased significantly at 3 hours of perfusion,and the hemoglobin level showed a downward trend at 1,3,5 and 6 hours.The number of white blood cells and platelets decreased at all time points.The blood oxygen saturation in the subnormothermic perfusion group remained stable at 95%-98%,and the levels of blood oxygen saturation,Na+,Ca2+,glucose and pH were stable,while the K+level first increased and then decreased.There was no significant difference in the levels of IgG and IgM before and after perfusion.The water content of brain tissue at the end of perfusion in the subnormothermic perfusion group was significantly higher than that in the positive control group(P<0.001).Nissl staining results showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex of the subnormothermic perfusion group maintained better morphological integrity,with no significant increase in enlarged and deformed cells.In the hippocampal CA1 region,there was a slight increase in enlarged and deformed cells,and a few cells with undamaged structures showed reduced cell size.In the hippocampal dentate gyrus,fewer granule neurons had compromised structural integrity,with increased cell edema.NEUN immunofluorescence staining showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex and hippocampal CA1 region of the subnormothermic perfusion group had better morphological states,with clear axons.The granule cells in the hippocampal dentate gyrus were well preserved,but the nuclei were less well protected.GFAP immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had sparser protrusions that were more tightly associated with neurons.Iba1 immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had thicker and fewer protrusions.Conclusions Compared with the positive control group,subnormothermic normoxic mechanical perfusion with genetically modified porcine erythrocyte perfusate increases brain tissue edema in cynomolgus monkeys,but better preserves the morphological integrity of neurons and glial cells.The protective effects may be related to the continuous oxygen and energy supply,maintenance of ion homeostasis and perfusate pH,reduced rejection,and low metabolic state of the whole brain.

Objective To investigate the relationship between the expression levels of serum multiple epithelial growth factor like domains 6(MEGF6),sterile alpha motif(SAM)domain,scr homology 3(SH3)domain and nuclear localization signals 1(SAMSN1)and acute lung injury(ALI)in patients with sepsis.Methods A total of 182 patients with sepsis(sepsis group)who were admitted to Huazhong University of Science and Technology Union Jiangnan Hospital from February 2021 to February 2024 were selected and divided into ALI group(n=60)and non-ALI group(n=122)according to whether they were complicated with ALI or not.80 healthy people who underwent physical examination during the same period were selected as the control group.Serum MEGF6 and SAMSN1 levels were detected by enzyme-linked immunosorbent assay(ELISA).Pearson correlation analysis was used to analyze the correlation between serum MEGF6,SAMSN1 and clinical indicators.Multivariate Logistic re-gression analysis was used to analyze the risk factors of sepsis complicated with ALI.The receiver operating characteristic(ROC)curve was used to analyze the value of serum MEGF6 and SAMSN1 in the evaluation of sepsis complicated with ALI.Results Serum MEGF6(37.52±7.87 ng/L)and SAMSN1(24.26±7.45 ng/L)in the sepsis group were lower than those in the control group(78.17±15.26 ng/L,62.31±14.12 ng/L),and the differences were statistically significant(t=28.397,28.477,all P<0.001).Serum MEGF6(29.22±7.64 ng/L)and SAMSN1(13.28±7.12 ng/L)in the ALI group were lower than those in the non-ALI group(41.60±7.90 ng/L,29.66±7.67 ng/L),and the differences were statistically significant(t=10.045,13.961,all P<0.05).Se-rum MEGF6 and SAMSN1 in patients with sepsis complicated with ALI were negatively correlated with APACHE II score and SOFA score(r=-0.811～-0.728,all P<0.05),and positively correlated with oxygenation index(OI)(r=0.689,0.640,all P<0.05).APACHE II score and SOFA score were risk factors for sepsis complicated with ALI.Serum MEGF6(OR=0.769,95%CI:0.612～0.965)and SAMSN1(OR=0.778,95%CI:0.617～0.980)were protective factors(Wald χ2=5.140,4.525,all P<0.05).The area under the curve(AUC)of serum MEGF6,SAMSN1 alone and combined in the evaluation of sepsis complicated with ALI were 0.840,0.811 and 0.903,respectively.The combined detection was greater than the single detection,and the differences were statistically significant(Z=4.602,4.318,all P<0.05).Conclusion The serum levels of MEGF6 and SAMSN1 in patients with sepsis are decreased,which are the influ-encing factors of sepsis complicated with ALI.The combined detection of the two has a high evaluation value for sepsis complicated with ALI.

Aim To explore the potential mechanisms underlying therapeutic effects of cordycepin in asthma by utilizing network pharmacology,molecular docking,and in vitro cellular validation.Methods The thera-peutic targets associated with asthma and the drug tar-gets of cordycepin were systematically identified through comprehensive database searches.An overlap analysis of the two gene sets was performed,followed by the construction of a protein-protein interaction(PPI)network and topological analysis to identify the core targets.The core targets were subjected to Kyoto Ency-clopedia of Genes and Genomes(KEGG)pathway a-nalysis and Gene Ontology(GO)enrichment analysis,and a drug-target-pathway network was constructed.To validate the interaction between cordycepin and core targets,molecular docking and molecular dynamics sim-ulations were conducted.Subsequently,the pharmaco-logical effects and underlying mechanisms of cordyce-pin were validated in vitro using Beas-2B cells,emplo-ying Cell Counting Kit-8(CCK-8)assay and quantita-tive real-time reverse transcription PCR(RT-qPCR).Results A total of 438 potential targets of cordycepin were identified,113 of which overlapped with asthma-related therapeutic targets.Topological analysis based on the PPI network revealed 22 core targets.Using KEGG enrichment analysis,165 significantly enriched pathways were identified,including the TNF and HIF-1 signaling pathways.Molecular docking analysis re-vealed high binding affinities between cordycepin and select core targets,which further corroborated by mo-lecular dynamics simulations.In vitro experiments showed that after cordycepin pretreatment,the upregu-lation of MAPK1,HIF1A,MTOR,MYC,IL10,and JUN mRNA was significantly rescued in HDM-stimulated Beas-2B cells.Conclusions Cordycepin exerts anti-asthmatic effects by targeting MAPK1 and other key molecules,thereby providing a scientific foundation for its further development and clinical application.