Objective To explore the feasibility and safety of non-anticoagulation veno-venous extracorporeal membrane oxygenation(VV-ECMO)in patients with severe chest trauma.Methods A retrospective cohort study method was used.A total of 19 patients with severe chest trauma who received VV-ECMO with a delayed anticoagulation strategy at Zhengzhou Central Hospital Affiliated to Zhengzhou University from January 2018 to October 2021 were included in the delayed anticoagulation group,and 20 patients with severe chest trauma who received VV-ECMO with a non-anticoagulation strategy from November 2021 to October 2024 were included in the non-anticoagulation group.The overall clinical characteristics of the patients were statistically analyzed,including gender,age,injury severity score(ISS),acute physiology and chronic health evaluationⅡ(APACHEⅡ),reason for VV-ECMO,use of vasoactive drugs,oxygenation index(PaO2/FiO2),and interval from injury to VV-ECMO.The primary outcomes were hemorrhagic and thrombotic complications.The secondary outcomes were blood transfusion during VV-ECMO,VV-ECMO time,mechanical ventilation time,intensive care unit(ICU)length of stay,and 28-day mortality.Results There was no significant difference in gender,age,ISS score,APACHEⅡscore,reason for VV-ECMO,use of vasoactive drugs,PaO2/FiO2,and interval from injury to VV-ECMO between the non-anticoagulation group and the delayed anticoagulation group.There was no significant difference in overall incidence of hemorrhagic and thrombotic between the two groups[incidence of hemorrhagic complications:15.0%(3/20)vs.31.6%(6/19),incidence of thrombotic:15.0%(3/20)vs.5.3%(1/19),both P>0.05].The infusion rate of 4 or more paked red blood cell(PRBC)within 24 hours during VV-ECMO in the non-anticoagulation group was significantly lower than that in the delayed anticoagulation group[5.0%(1/20)vs.31.6%(6/19),P<0.05].The amount of PRBC and platelet transfusion and the time on VV-ECMO in the non-anticoagulation group during VV-ECMO were significantly lower than those in the delayed anticoagulation group[PRBC(U):5.8±3.8 vs.8.1±3.1,platelets(U):1(0,1)vs.2(1,3),time on VV-ECMO(hours):71.55±24.37 vs.114.21±34.08,all P<0.05].There were no statistically significant differences in the amount of plasma and cryoprecipitate transfusion during VV-ECMO,mechanical ventilation time,ICU hospitalization time,and 28-day mortality between the two groups.Conclusion For patients with severe chest trauma receiving VV-ECMO withholding routine systemic anticoagulation did not result in thrombotic complications or higher mortality and required less PRBC and platelet transfusions.Non-anticoagulant VV-ECMO is safe and feasible for patients with severe chest trauma with high risk of bleeding.

Objective:To preliminarily explore whether sirtuin1 (SIRT1) activation can inhibit neuronal ferroptosis in rats after traumatic brain injury (TBI) by regulating hypoxia-inducible factor-1α (HIF-1α)-mediated glycolysis.Methods:(1) Six SD rats were randomly divided into sham-operated group and TBI group, with 3 rats in each group; TBI model in the TBI group was established by hydraulic impact method, and rats in the sham-operated group underwent same surgery without impact. Cortical tissues of the two groups were sent for tandem mass tag (TMT) labeled quantitative proteomics detection to analyze the differential expression proteome; Kyoto encyclopedia of genes and genomes (KEGG) and gene set enrichment analysis (GSEA) were used to detect pathway enrichment of the screened differential proteins. (2) Twelve SD rats were randomly divided into sham-operated group and 1-day, 3-day and 7-day post-TBI groups, with 3 rats in each group. Treatment methods were the same as above; Western blotting was used to detect SIRT1 protein expression. (3) Forty-eight rats were randomly divided into sham-operated group, TBI group, TBI+vehicle group and TBI+SIRT1 agonist group, with 12 rats in each group; rats in the sham-operated group and TBI group accepted treatment as above; rats in the TBI+SIRT1 agonist group were intraperitoneally injected with SRT1720 (dissolved in ≤ 5% dimethyl sulfoxide, at a dose of 20 mg/kg) within 30 minutes after modeling, twice a day (with an interval of 12 hours); and rats in the TBI+vehicle group were injected with same dose of dimethyl sulfoxide at the same time. One d after modeling, neurological deficit was assessed using modified Neurological severity score (mNSS), brain water content was measured by dry-wet weight method, histopathological changes in the cortical lesions were observed by HE staining, mitochondrial ultrastructure was examined by transmission electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the brain tissues were detected by colorimetry, and protein expressions of SIRT1, HIF-1α (key protein in the glycolytic pathway), glutathione peroxidase 4 (GPX4, key protein in the ferroptosis pathway), and acyl-CoA synthetase long-chain family member 4 (ACSL4, key protein in the ferroptosis pathway) were evaluated by Western blotting.Results:(1) KEGG analysis revealed that the glycolysis pathway and HIF-1 signaling pathway were obviously enriched in the cortical tissues of rats in the TBI group compared with the sham-operated group; GSEA showed that the HIF-1 signaling pathway (mmu04066) and ferroptosis pathway (mmu04216) gene sets in the cortical tissues of rats in the TBI group exhibited enrichment trends compared with those in the sham-operated group. (2) Compared with the sham-operated group, the 1-day, 3-day, and 7-day post-TBI groups had significantly decreased SIRT1 protein expression ( P<0.05), with the most prominent decline in 1-day post-TBI group. (3) Compared with the TBI+vehicle group, rats in the TBI+SIRT1 agonist group showed significantly reduced mNSS score and brain tissue water content (9.83±1.17 vs. 7.66±1.21; [83.62±0.91]% vs. [80.09±0.68]%, P<0.05). HE staining indicated clearer structure of the cortical area at the injury sites, and improved neuron morphology in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group; and transmission electron microscopy showed reduced mitochondrial shrinkage and partial restoration of cristae structures in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group. Compared with the TBI+vehicle group, the TBI+SIRT1 agonist group exhibited significantly decreased MDA content ([62.72±9.20] nmol/g vs. [39.34±3.48] nmol/g), increased SOD activity ([1.95±0.23] U/mg vs. [2.48±0.14] U/mg), elevated GPX4 protein expression (0.37±0.04 vs. 0.46±0.03), and decreased HIF-1α and ACSL4 protein expressions (1.16±0.15 vs. 0.81±0.12; 1.14±0.06 vs. 1.29±0.04), with significant differences ( P<0.05). Conclusion:SIRT1 activation can exert neuroprotective effect by inhibiting HIF-1α-mediated glycolysis and reducing neuronal ferroptosis after TBI.

Objective:To measure and analyze the occlusal force and contact in children with mixed dentition,and to preliminarily provide baseline data on the occlusion of individual normal occlusion children with mixed dentition.Methods:A cross-sectional study was conducted,including 20 children with mixed dentition and individual normal occlusion,consisting of 12 boys and 8 girls,aged 6.5-9.8 years.The Dental Prescale Ⅱ occlusal analysis system was used to measure the occlusal force and contact at the intercuspal position,including the maximum occlusal force(N)and the occlusal contact area(mm2)of the entire dentition,and the left and right sides,average occlusal pressure(MPa),maximum occlusal pressure(MPa),and to determine the position of the center of occlusal force.The gender differences in maximum occlusal force,average occlusal pressure,and occlusal contact area were ana-lyzed,the bilateral symmetry of occlusion in children with mixed dentition and individual normal occlu-sion was compared,and the correlation between occlusal data and age,height,weight,and body mass index(BMI)was analyzed.Results:(1)The average maximum occlusal force of the entire dentition in the 20 children with mixed dentition at the intercuspal position was(869.18±106.64)N,the average occlusal contact area was(25.19±2.89)mm2,the average occlusal pressure was(34.37±5.98)MPa,and the maximum occlusal pressure M(P25,P75)was 120(120,120)MPa;(2)There was no statistically significant difference in the maximum occlusal force,average occlusal pressure,maximum occlusal pressure,and occlusal contact area between the left and right sides(P＞0.05);(3)At the intercuspal position,the average occlusal contact area for 12 boys and 8 girls was(26.71±3.91)mm2and(21.62±3.08)mm2 respectively,and the average maximum occlusal force was(911.92±145.05)N and(769.47±116.45)N respectively,with statistically significant differences(P＜0.05),while there was no statistically significant difference in the average occlusal pressure between boys and girls(P＞0.05);(4)The maximum occlusal force at the intercuspal position was weakly correlated with age(r=0.219,P=0.046),and strongly positively correlated with the occlusal contact area(r=0.949,P＜0.001),while the average occlusal pressure,maximum occlusal pressure,and occlusal contact area were not correlated with age,height,weight,or BMI;(5)The center of occlusal force in the 20 children with mixed dentition and individual normal occlusion was located in the molar region,with 7 children having the maximum occlusal pressure point only in the first permanent molar region,10 children having it in both the deciduous molar region and the first permanent molar region,and 3 children having it only in the deciduous molar region.Conclusion:In children with mixed dentition and individual normal occlusion,the maximum occlusal force,occlusal contact area,average occlusal pressure,and maximum occlusal pressure at the intercuspal position show good bilateral symmetry;there are gender differences in the maximum occlusal force and occlusal contact area,with boys having greater values than girls;the maxi-mum occlusal force is positively correlated with the occlusal contact area.

Objective:This study aimed to investigate the morphological and functional changes of jejunal and ileal epithelium in aged mice using 3D organoid culture, to elucidate the age-related alterations in epithelial function across different regions of the intestine.Methods:The C57BL/6J male mice were divided into two groups: a young group(5 months)and an old group(24 months).Epithelial tissues from the jejunum and ileum were harvested, and the jejunal and ileal epithelial crypt units were separated for 3D organoid culture in vitro.Histological analyses, including H&E and PAS staining, immunofluorescence staining, and quantitative PCR, were employed to assess changes in crypt depth, villi length, goblet cell numbers, inflammation, and mucosal barrier function in the aged mice.Results:With aging, the crypt of jejunum became deeper and the ratio of villi length to crypt depth decreased, while the villi length of ileum became longer and the ratio of villi length to crypt depth increased.The rate of organoid formation in jejunal and ileal crypts and the number of crypto-like regions of organoid formation were significantly decreased in the aged mice, along with a decrease in the expression of the stem cell proliferation marker Ki67.The inflammation of the jejunal and ileal epithelium increased in the aged mice, and the mechanical barrier function of the epithelial mucosa was enhanced.Conclusions:In aged mice, both the jejunal and ileal epithelium exhibited similar changes in stem cell proliferation, inflammation, and mucosal mechanical barrier function, though the changes in villus length and crypt depth ratio were opposite.This study provides a foundation for further investigation of the differences in jejunal and ileal epithelial function and its potential mechanisms during aging.

Objective:To investigate the molecular pathogenic mechanism of venous thrombosis caused by heterozygous missense mutations in two protein C (PROC) genes through laboratory phenotype analysis, genetic mutation analysis, and in vitro expression experiments.Methods:Two probands presented with venous thromboembolism at the First Affiliated Hospital of Wenzhou Medical University. Clinical data and blood samples were collected from the probands and their family members to evaluate the plasma protein C (PC) activity (PC∶A), PC antigen (PC∶Ag) levels, and other relevant coagulation parameters. The anticoagulant capacity was assessed using the thrombin generation test (TGT). The mutation sites of the PROC gene were identified using direct DNA sequencing. Bioinformatics software was used to analyze the conservation and pathogenicity of the mutated gene. PyMOL software was used for the analysis of the protein three-dimensional models and interactions between mutated amino acids. Wild-type and two mutant expression vectors were constructed and HEK293T cells were transiently transfected. Total cellular RNA was extracted from positively transfected cells to investigate the transcriptional levels of the mutant PROC gene. Enzyme-linked immunosorbent assay, Western blot, and cellular immunofluorescence assays were used to investigate the translation levels of the mutant PROC protein.Results:Probands 1 and 2 exhibited PC∶A levels of 35% and 40% and PC∶Ag levels of 44% and 39%, with increasing D-dimer levels to 4.42 mg/L and 0.83 mg/L, respectively. Meanwhile, other coagulation parameters revealed no significant abnormalities. TGT demonstrated impaired anticoagulant function in both proband witnesses and their familial PC carriers. Sequencing analysis revealed heterozygous missense mutations c. 833T>C (p. Leu278Pro) in proband 1 and c. 1330T>C (p. Trp444Arg) in proband 2 within exon 9 of the PROC gene. Conservation analysis revealed that Leu278 and Trp444 were highly conserved across homologous species. Pathogenicity analysis indicated that both p. Leu278Pro and p. Trp444Arg mutations are deleterious. Protein modeling analysis demonstrated that both mutations induce structural alterations in the protein. In vitro expression experiments revealed that compared with the wild-type, both p. Leu278Pro and p. Trp444Arg mutations showed no significant differences in the mRNA expression level of the PC protein. However, both mutations caused significantly lower PC∶Ag content and protein expression levels in the cell culture supernatant compared with the wild-type, whereas higher levels were observed in the cell culture lysate. This indicates the association of both mutations with the secretion function of the PC protein.Conclusion:The heterozygous missense mutations p. Leu278Pro and p. Trp444Arg in exon 9 of the PROC gene in both probands are associated with decreased PC levels.

Objective:To develop an artificial intelligence-based model for automated recognition of serum indices using machine vision and deep learning.Methods:This study was a cross-sectional study.Serum sample images were collected fromWest China Hospital of Sichuan University from September 21, 2020 to January 20, 2023 using the imaging device of the fully automated sample pre-processing system. A computer random number generator was used to randomly select one whole hour each day, and all serum sample images processed within that hour were included. After excluding samples with unqualified images and missing serum index results, a total of 5, 534 samples were included. These were divided into a training set and a test set in an 8∶2 ratio using Python random shuffle function, and 4, 458 samples were in the training set and 1, 076 samples were in the test set. After manual inspection, the serum regions were annotated using the MATLAB Image Labeler tool and converted into YOLO format, and a YOLO v5-based model was constructed for automatic serum region extraction. The actual values of lipemia index (L-index), hemolysis index (H-index), and icterus index (I-index) were measured using the automatic biochemical analyzerwith matched reagent kits. A serum index regression model was constructed based on the MobileNet v2 network using the PyTorch 1.10.0 framework. The grading performance of the model was evaluated using accuracy, Kappa coefficient, sensitivity, specificity, positive predictive value, and negative predictive value. Regression performance was assessed using root mean square error (RMSE), mean absolute error (MAE), coefficient of determination (R2), and Bland-Altman analysis.Results:The overall accuracy rates for grading L-index, H-index, and I-index were 98.88%, 95.26%, and 92.47%, respectively, with Kappa coefficients of 0.72, 0.72, and 0.59. For L-index, MAE was 5.11, RMSE was 9.77, and R2 was 0.78. For H-index, MAE was 5.18, RMSE was 8.99, and R2 was 0.89. For I-index, MAE was 1.13, RMSE was 3.01, and R2 was 0.71. Bland-Altman analysis showed that 95.5%, 95.1%, and 95.7% of the data points fell within the consistency intervals for L-index, H-index, and I-index, respectively.Conclusion:The study developed an artificial intelligence-based serum index regression modelto estimate serum indices with high efficiency and accuracy. It shows great potential for reducing laboratory costs, improving clinical testing efficiency, and promoting intelligent development in laboratory medicine.

Objective:To analyze the gene mutations of 18 patients with plasminogen (PLG) deficiency and to explore the clinical manifestations caused by PLG gene mutations.Methods:This study belongs to observational study-descriptive study: case series.Clinical data from 18 patients with PLG deficiency admitted to the First Affiliated Hospital of Wenzhou Medical University from January 1st, 2021 to May 31st, 2025 were collected. The age ranged from 16 to 70 years old, with an average of 48 years old. Among them, there were 10 males and 8 females. Anticoagulant blood samples were taken before treatment to measure and analyze plasminogen activity (PLG:A), plasminogen antigen (PLG:Ag), protein C activity, protein S activity, fibrinogen, antithrombin activity, D-dimer, and fibrin (fibrinogen) degradation products. PCR direct sequencing was used to analyze the 19 exons and flanking sequences of the PLG gene in these patients, and reverse sequencing was employed to verify the suspected mutations.Results:For the 18 patients, cranial MRI showed fresh cerebral infarction lesions, and PLG:A levels ranged from 19% to 67%, while no other lab indicators showed significant abnormalities, all presenting with dysplasminogenemia. Genetic analysis revealed five types of PLG gene mutations: c.1858G>A (p.Ala620Thr) heterozygous mutation, c.1858G>A (p.Ala620Thr) homozygous mutation, c.398A>G (p.His133Arg) heterozygous mutation, c.2108G>A (p.Gly703Asp) heterozygous mutation, and c.1702G>A (p.Gly568Arg) heterozygous mutation. Among the above, the c.1858G>A heterozygous mutation was the most common, and c.398A>G and c.1702G>A were identified for the first time.Conclusion:Patients with plasminogen deficiency caused by PLG gene defects are prone to occur cerebral infarction events, which may be related to impaired fibrinolytic function due to PLG gene mutations.

Objective:To observe the clinical features and treatment outcomes of patients with intraocular foreign bodies with endophthalmitis, and analyze the prognostic factors affecting the anatomic and visual outcomes of patients.Methods:A retrospective clinical study. A total of 1 704 patients (1 704 eyes) with intraocular foreign body at Eye Hospital, Wenzhou Medical University from January 2015 to June 2024 were included in this study. Endophthalmitis was diagnosed in 263 eyes (15.4%, 263/1 704). Patients who lost follow-up in our hospital after surgery were excluded, 155 patients with 155 eyes were finally included in the study. Uncorrected visual acuity (UCVA) examination was performed before operation. Best corrected visual acuity (BCVA) examination was performed both after the first stage debridement and during follow-up. The visual acuity test is performed using a standard logarithmic visual acuity chart, which is statistically converted to logarithm of the minimum angle of resolution (logMAR) visual acuity. Demographic characteristics (gender, age), trauma characteristics (time of injury, occupation characteristics, nature of foreign body), anatomical injury (wound zoning, nature of infection, etc.), clinical treatment (interval between operation and injury, rate of second operation, etc.) and outcome (vision outcome, complications, anatomic outcome, etc.) were recorded. Prophylactic intravitreous injection of 10 mg/ml of cefazolin sodium 0.1 ml (including 1 mg of cefazolin sodium) was given on the basis of perioperative systemic administration of cefazolin sodium from 2022. Anatomical outcomes included anatomical reduction, silicone oil-dependent, and ophthalmectomy. The visual outcomes of the patients were categorized into three groups based on the best-corrected visual acuity at the final follow-up: visual acuity worse than 0.05, visual acuity between 0.05 and 0.3, and visual acuity better than 0.3. Generalized linear mixed model (GLMM) was used to analyze the correlation between the timing of treatment, nature of foreign body, nature of infection, number of operations, location of injury and the anatomic and visual outcomes of patients.Results:Of 155 patients, 149 were males and 6 were females, mean age was (45.7±12.9) years, patients with monocular injury. Magnetic, non-magnetic, unidentified metal and vegetable, mineral, animal and unidentified foreign bodies were 102 (65.8%, 102/155), 2 (1.3%, 2/155), 28 (18.1%, 28/155), 1 (0.6%, 1/155), 12 (7.7%, 12/155), 7 (4.5%, 7/155), 3 (1.9%, 3/155) cases, respectively. The time between injury and removal of foreign body was (98.1±359.5) h. The foreign bodies were removed in 136 eyes (87.2%, 136/155) in the primary surgery, 67 cases combined with debridement and suture, 68 cases combined with pars plana vitrectomy (PPV), and 1 case suffered ophthalmectomy. The slide and culture results revealed that the eyes positive for bacteria and those positive for a mixed infection of bacteria and fungi were 80 (51.2%, 80/155) and 2 (1.3%, 2/155) eyes, respectively; 73 eyes (46.8%, 73/155) were negative. Among the 80 eyes positive for bacteria, staphylococcus epidermidis and bacillus cereus were found in 26 (32.5%, 26/80) and 23 (28.8%, 23/80) eyes, respectively. Drug sensitivity testing indicated that vancomycin, gentamicin and amikacin had low drug resistance (1.79%, 6.67%, 0.0%, respectively). The mean preoperative logMAR UCVA was 1.67±0.79. In the outcome of visual function, 78, 26 and 51 patients with visual acuity <0.05, 0.05-0.3, >0.3, respectively. At the last follow-up, there were 56 cases (36.1%, 56/155) of silicone oil dependence, 93 cases (60%, 93/155) of anatomic reduction, and 6 cases (3.9%, 6/155) suffered ophthalmectomy, the mean intraocular pressure was (13.6±6.1) mm Hg (1 mm Hg= 0.133 kPa). Preoperative visual acuity was strongly correlated with visual outcomes ( F=6.896, P=0.001). Preoperative visual acuity ( F=5.310, P=0.023) and surgical method ( F=20.448, P<0.001) were closely related to the anatomical outcome, while age, treatment time, foreign body nature, wound zoning, infection nature, and foreign body removal time had no statistical correlation with the anatomic and functional outcome ( P>0.05). During 2015 to 2024, the incidence of intraocular foreign body-related endophthalmitis was 12.5%-22.7%, which showed a fluctuating upward trend. The incidence of endophthalmitis increased during 2022 to 2024 compared with the period from 2019 to 2021, but no statistically difference was found ( χ2=3.856, P=0.05). Conclusions:The incidence of intraocular foreign body related endophthalmitis was 15.4%. Staphylococcus epidermidis and Bacillus cereus are the first and second pathogenic bacteria. The incidence of endophthalmitis is not significantly reduced with intravitreal injection of cefazolin sodium. Preoperative UCVA and surgical method were closely related to the anatomic outcome of patients.

This study aims to investigate the effects of aflatoxin B1(AFB1)on the infection and replication of porcine delta coronavirus(PDCoV).Porcine small intestinal epithelial cells(IPEC-J2),porcine kidney cells(LLC-PK)and swine testis cells(ST)were used as models,and CCK-8 method was used to detect the safe mass concentration range of AFB1 on the three cell lines.Each cell line was divided into the blank control group,AFB1 treatment group,PDCoV treatment group and AFB1 and the PDCoV co-treatment group.The cells were treated with safe mass concentration of AFB1 for 12 h,and then treated with PDCoV for 20 h.Total RNA and total protein were extrac-ted from the cells.The effects of AFB1 on PDCoV infection and replication were detected by qPCR,Western blot and cell immunofluorescence tests.The results showed that compared with the PDCoV treatment group,the mRNA and N protein of viral S protein in the AFB1 and PDCoV co-treatment group were significantly increased(P＜0.05),and a significantly higher fluorescence of PDCoV N protein is also visibly present in the co-treatment group,and there was a significant difference between the two groups.The results showed that AFB1 could promote the infection and replication of PDCoV.It provides important data that AFB1 can promote the infection replication of PDCoV and provides a new idea for the prevention and treatment of PDCoV.

Objective:To investigate the current status of teaching behavior of clinical nursing teachers and explore the mediation effect of organizational commitment between their work engagement and teaching behavior.Methods:This study included a total of 1 736 clinical nursing teachers who worked in grade A tertiary hospitals in Beijing City, Shanghai City, Henan Province, Hubei Province, Hebei Province, and Chongqing City from May 2022 to November 2022. Data were collected using a general information questionnaire, the Effective Teaching Behavior Scale, the Work Engagement Scale, and the Organizational Commitment Scale. Statistical description and correlation analysis were performed using SPSS 25.0 software, and mediation effect was analyzed using AMOS 25.0 software.Results:The total mean score of effective teaching behavior of 1 599 clinical nursing teachers was (4.19±0.66), the total mean score of work engagement was (5.09±1.36), and the total mean score of organizational commitment was (3.93±0.72). Pearson correlation analysis showed that work engagement and organizational commitment were positively correlated with teaching behavior ( r=0.560, 0.519, both P<0.01). Organizational commitment partially mediated the relationship between work engagement and teaching behavior, with a mediation effect value of 0.27, and the mediation effect accounted for 49.10% of the total effect. Conclusions:Work engagement is a direct predictor of the teaching behavior of clinical nursing teachers, and organizational commitment partially mediates the relationship between work engagement and teaching behavior. Administrators can promote the work engagement of clinical nursing teachers by adopting effective interventions to increase the level of organizational commitment, which in turn enhances teaching behavior.