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.

Currently, the quality control of traditional Chinese medicine (TCM) encounters substantial technical challenges, necessitating the urgent development of innovative evaluation methods for key quality attributes. This study proposes the construction of a quality assurance system centered on key quality attributes, with systems biology and chemical biology serving as its core components: utilizing multi-omics technologies to systematically analyze the core functional modules of TCM, integrating pharmacological approaches to precisely identify key bioactive components; applying chemical biology to elucidate the interaction mechanisms between components and their targets, while screening for quality markers associated with specific functions and clinical indications; establishing an integrated quality control system based on quality markers and biological potency to ensure comprehensive quality management from raw herb materials through to finished products. Additionally, it emphasizes the formulation of compatibility rules grounded in TCM theory, integrates clinical practice and multi-dimensional value assessments, and establishes a novel quality evaluation model with distinct TCM characteristics.This offers a comprehensive and systematic approach to ensuring the safety and efficacy of TCM while facilitating the high-quality development of the TCM industry.

Objective To systematically evaluate the similarities between six-gene-modified pig red blood cells and human red blood cells in multiple dimensions, including cell morphology, hematological parameters, microbial safety and serological compatibility, and to verify the feasibility of using six-gene-modified pig red blood cells as an alternative for blood transfusion in cases of traumatic hemorrhage. Methods Blood samples from O-type human donor, six-gene-modified O-type pig and wild-type O-type pig were collected. The pig gene-modified phenotype was identified by flow cytometry. The morphology was observed using field emission scanning electron microscopy and the diameter was measured. Hematological tests, including blood routine, osmotic fragility test, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) content and partial pressure of oxygen at 50% hemoglobin saturation (P50), were conducted to assess the deformability of red blood cells. Real-time fluorescent quantitative polymerase chain reaction was used to screen for zoonotic viruses, and cross-matching was performed using column gel method, condensation amine method and saline tube method. Results The six-gene-modified pig red blood cells successfully knocked out the α-Gal, Neu5Gc and Sda antigens and expressed hCD55. Wild-type pig red blood cells expressed these three antigens but did not express human complement regulatory proteins. All three types of red blood cells presented a biconcave discoid morphology. The diameter of O-type human red blood cells was larger than that of the six-gene-modified porcine red blood cells and wild-type pig red blood cells (all P < 0.05). The average red blood cell volume and average red blood cell hemoglobin of O-type humans were higher than those of the six-gene-modified pig red blood cells, and the distribution width-variable coefficient of O-type human red blood cells was lower than that of the six-gene-modified pig red blood cells (all P < 0.05). There were no statistically significant differences in the permeability fragility, ATP content, 2,3-DPG content, P50 and red blood cell deformability indicators of the three types of red blood cells (all P > 0.05). The screening for zoonotic viruses in human and animal cells of the six-gene-modified pigs and wild-type pigs was negative. The cross-matching results showed that in the column gel method and condensation amine method, the primary and secondary sides of the six-gene-modified pig red blood cells did not agglutinate with human red blood cells, while the wild-type pig red blood cells agglutinated with human red blood cells in all cross-matching methods. Conclusions The six-gene-modified pig red blood cells are highly similar to human red blood cells in terms of morphology, key hematological parameters, energy metabolism, oxygen-carrying capacity, mechanical properties, microbial safety and serological compatibility, providing an experimental basis for the clinical transformation of xenotransfusion.

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.

Objective:To explore the effects of 4C management mode on rehabilitation and quality of life in patients with acute heart failure(AHF)in cardiovascular intensive care unit(CCU).Methods:This randomized controlled study enrolled 110 patients with AHF admitted in CCU of Guangdong Provincial Hospital of Traditional Chinese Medicine between February 2021 and February 2023.Patients were randomly divided into intervention group(n=55)and control group(n=55).Patients in the control group received routine intervention,while those in the inter-vention group received additional 4C management mode,both groups were intervened for 3 months.The indexes of cardiac function,self-efficacy,exercise tolerance,blood oxygen saturation(SpO2),mental state and quality of life were compared between the two groups.Results:Compared to those in the control group after intervention,patients in the intervention group had significantly lower interventricular septal thickness(IVST)[(9.20±0.10)mm vs.(10.30±0.17)mm,P＜0.001],left ventricular end-systolic volume index(LVESVI)[(35.21±0.74)ml/m2 vs.(39.70±1.09)ml/m2,P＜0.001],left ventricular end-diastolic volume index(LVED VI)[(45.35±0.71)ml/m2 vs.(56.76±1.48)ml/m2,P＜0.001],scores of self-rating anxiety scale(SAS)[(32.10±1.57)points vs.(45.36±1.94)points,P＜0.001],self-rating depression scale(SDS)[(35.03±1.52)points vs.(47.70±2.12)points,P＜0.001]and Minnesota living with heart failure questionnaire(MLHFQ)[(51.27±2.85)points vs.(56.46±3.53)points,P＜0.001],and significantly higher score of general self-efficacy scale(GSES)[(28.71±3.84)points vs.(20.34±1.56)points,P＜0.001],6 min walking distance(6MWD)[(428.53±5.50)m vs.(362.14±6.18)m,P＜0.001]and SpO2[(99.11±0.87)％vs.(94.23±0.73)％,P＜0.001].Conclusion:4C management mode may improve the cardiac function,self-efficacy,mental state and quality of life in patients with acute heart failure in CCU.

Purpose To achieve intelligent detection of acute pulmonary embolism(APE)in CT pulmonary angiography based on the Res2Net attention mechanism network.Materials and Methods Retrospectively included patients with suspected of APE who underwent CT pulmonary angiography examination and were diagnosed as APE at the First Affiliated Hospital of Henan University of Chinese Medicine from February 2015 to May 2023.The dataset was randomly divided into training,validation and test set in a ratio of 7∶2∶1.The model was trained based on the Res2Net network,combined with atrous spatial pyramid pooling and attention mechanism modules,and was performed five-fold cross internal validation.Using the area under the receiver operator characteristic curve,sensitivity and specificity to assess the diagnostic performance of the model.Dice similarity coefficient,precision and intersection over union(IOU)were used to assess the segmentation performance of thrombus on the test set and plot the corresponding curves.The performance of the Res2Net attention mechanism network was compared with the classic U-Net and CE-Net model.Results A total of 303 patients with APE were included in this study.There were 212,61 and 30 cases in the training set,validation set and test set,respectively.The model's area under the curve was 0.95,sensitivity was 0.90,specificity was 1.00,Dice similarity coefficient was 0.86,precision was 0.90,Pos-IOU was 0.78 and Neg-IOU was 1.00,respectively.The parameter curves and radar chart showed that the Res2Net attention mechanism network performed better than the U-Net and CE-Net models.The visualization results of the segmentation comparison showed that the Res2Net attention mechanism network achieved higher precision in segmenting pulmonary artery thrombus.Conclusion The Res2Net attention mechanism network has good performance for detection of APE.

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.

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 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.