Objective The aim of this study was to clarify the role and mechanism of Pseudomonas aeruginosa vaccine subunit OprI in the fusion protein vaccine rePO(PcrV-OprI).Methods The in vitro stability of rePO,PcrV and OprI at 4 ℃,25 ℃,and 37 ℃ was examined.After immunizing mice with rePO,OprI and PcrV,respectively,the specific antibody potency in serum and the proportion of cells secreting IFN-γ and IL-4 in the spleen were examined;Additionally,detection of the levels of protein uptake by DC2.4 cells in vitro using laser confocal microscopy and flow cytometry,and their ability to promote the maturation of mouse bone marrow-derived dendritic cells(BMDC).Results The heat stability of fusion protein rePO was significantly better than that of PcrV.The induced anti-PcrV IgG and anti-OprI IgG potency of rePO was significantly higher than that of monomeric PcrV and OprI.Additionally,the number of cells secreting IFN-γ and IL-4 induced by immunization with rePO was significantly higher than that of PcrV and OprI.The uptake rate of fusion protein rePO by DC2.4 cells was significantly higher than that of PcrV and OprI.Furthermore,rePO promoted the maturation of mouse BMDC more effectively than PcrV and OprI.Conclusion OprI in the fusion protein rePO can significantly improve its thermal stability and immunogenicity,which lays the foundation for the successful development of Pseudomonas aeruginosa vaccine.

A 36-year-old male developed unconsciousness and no response to voice stimuli after taking approximately 2 050 mg felodipine (the specific time was unknown). Two hours later, he was sent to the department of emergency by his family and admitted to the hospital. His vital signs showed body temperature 35.1 ℃, pulse 148 times/min, respiration 32 times/min, and blood pressure 65/34 mmHg. Acute drug poisoning, acute toxic cardiomyopathy, acute toxic shock, acute type Ⅱ respiratory failure, acute toxic encephalopathy, and acute renal failure were diagnosed based on the patient′s clinical manifestations combined with laboratory tests results, cardiac ultrasound, chest and abdominal CT scans. Endotracheal intubation connected to a ventilator for invasive assisted ventilation, pressure boosting, and fluid resuscitation were given. At the same time, repeated gastric lavage and enema were performed to remove toxins. Blood perfusion was intermittently and repeatedly administered, and continuous renal replacement therapy was used. The blood concentration of felodipine was 1 298 μg/L at 2 hours after admission, and cardiac arrest occurred at 4 hours. Venous-arterial extracorporeal membrane oxygenation (V-A ECMO) treatment was administered immediately. After 48 hours of ECMO operation, sedatives were discontinued and the patient′s consciousness was improved after 4 hours. On the 5th day of ECMO treatment, his heart rate was 72 beats per minute, and blood pressure was 127/65 mmHg. The blood concentration of felodipine decreased to 2 μg/L. The patient′s vital signs were significantly improved and ECMO supportive treatment was withdrawn. After 26 days of hospitalization, the patient recovered and was discharged.

Since the outbreak of COVID-19, many skin diseases related to or secondary to COVID-19 have been reported globally, and the infection can trigger or exacerbate skin diseases and other systemic symptoms. The review aims to discuss the manifestations and management of severe skin diseases following COVID-19, such as systemic lupus erythematosus, severe psoriasis, pemphigus, dermatomyositis, and Beh?et's syndrome, with the goal of improving the diagnosis and treatment of severe skin diseases after COVID-19.

Objective:To construct a risk prediction model for Vaginal Intraepithelial Neoplasia Grade 2 or Worse(VaIN 2+)lesions,and to establish a nomogram for individual diagnosis of VaIN 2+and risk stratification,so as to provide guidance for the treatment of vaginal lesions.Methods:A total of 248 women diagnosed with VaIN through colposcopic biopsy at the Center for Gynecologic and Cervical Diseases,First Affiliated Hospital of Nanjing Medical University,from January 2021 to January 2024 were included in this study.Based on the gold standard established by histological and pathological findings,these patients were categorized into a lower VaIN 2 group and a VaIN 2+group.Univariate comparative analysis was performed on the two groups.Multivariate Logis-tic regression analysis was used to determine the risk factors of VaIN 2+and to construct a diagnostic model.The nomogram model was established by using R Studio software.The discrimination,calibration and clinical practical value of the model were evaluated by the area under the receiver operating characteristic(ROC)curve and cali-bration curve.Results:Univariate analysis identified that HPV type,cervical lesion grade,acetowhite change,vagi-nal lesion duration,vaginal lesion location,and cervical lesion duration as influencing factors for diagnosing VaIN 2+(P<0.1).Multivariate binary Logistic regression analysis indicated that HPV16/18 positivity,cervical lesion grade≥CIN 2,thick acetowhite change,vaginal lesion duration≥5 years,and vaginal lesion location at the upper 1/3 of the vagina were independent risk factors for diagnosing VaIN 2+(OR>1,P<0.05),while cervical lesion duration<3 years was a protective factor(OR<1,P<0.05),with acetowhite change having the greatest impact(OR4.54).A regression model was established based on the multivariate binary Logistic regression analysis,with an AUC of 0.813.A nomogram model was constructed and internally validated,yielding a consistency index(C-index)of 0.81.Patients were stratified into risk groups using the X-tile software,with higher total scores indi-cating a greater risk of developing VaIN 2+.Conclusions:The nomogram model constructed in this study can in-dividually predict the risk of VaIN 2+lesions in patients,with high accuracy and clinical practicability.

In current clinical practice, various dermal templates and skin substitutes are used to enhance wound healing. However, the role of wound commensal microbiome in regulating scaffold performance and the healing process remains unclear. In this study, we investigated the influence of both natural and synthetic scaffolds on the wound commensal microbiome and wound repair in three distinct models including diabetic wounds, burn injuries, and germ-free (GF) wounds. Remarkably, synthetic electrospun polycaprolactone (PCL) scaffolds were observed to positively promote microbiome diversity, leading to enhanced diabetic wound healing compared to the natural scaffolds Integra® (INT) and MatriDerm® (MAD). In contrast, both natural and synthetic scaffolds exhibited comparable effects on the diversity of the microbiome and the healing of burn injuries. In GF wounds with no detectable microorganisms, a reversed healing rate was noted showing natural scaffold (MAD) accelerated wound repair compared to the open or the synthetic scaffold (PCL) treatment. Furthermore, the response of the wound commensal microbiome to PCL scaffolds appears pivotal in promoting anti-inflammatory factors during diabetic wound healing. Our results emphasize that the wound commensal microbiome, mediated by different scaffolds plays an important role in the wound healing process.

Objective:To analyze the clinical and endoscopic features of elderly patients with early gastric neuroendocrine neoplasms(g-NEN)to provide insights for clinical diagnosis and treatment.Methods:Seven cases of early-stage g-NEN in elderly patients admitted to Beijing Hospital from May 2020 to October 2024 were reported.Information including patient demographics, medical history, laboratory findings, endoscopic features, and pathological results was summarized an analyzed.Meanwhile, a literature review was conducted on g-NEN cases reported domestically and internationally from the establishment of databases to April 2025, with a comparative analysis of baseline characteristics, background mucosal conditions, and pathological diagnoses.Results:A total of 7 elderly patients with early g-NEN were included, including 3 males and 4 females, with an average age of 67 years at diagnosis.Endoscopic and pathological features: endoscopic lesions were mainly located on the greater curvature of the stomach, mostly slightly elevated, with 4/7(57.1%)being single lesions and the others accompanied by multiple polyps.The size of gastric neuroendocrine tumor(g-NET) lesions ranged from 1 mm to 19 mm, with a median Kyoto gastritis score of 3.In terms of pathological grading, there were 6 cases of G1 and 1 case of gastric neuroendocrine carcinoma(g-NEC). All 7 patients underwent endoscopic submucosal dissection(ESD). The follow-up period after surgery ranged from 3 months to 4 years, and no tumor was found at the vertical and horizontal margins of all lesions on pathology.A literature review of 32 reported cases showed that 40.6%(13/32)of cases exhibited background mucosal atrophy or intestinal metaplasia, with positive rates of chromogranin A(CgA) and synapsin(Syn)reaching 95.8%(23/24) and 100.0%(25/25), respectively.Notably, elderly patients with g-NEC demonstrated aggressive invasiveness, with a metastasis rate of 62.5%(5/8).Conclusions:Elderly patients with g-NEN exhibit high heterogeneity and significant differences in invasiveness, necessitating heightened clinical vigilance.Precise management should integrate background mucosal status, Kyoto gastritis score, pathological biomarkers, and imaging techniques.Blue laser imaging with magnifying endoscopy(BLI-ME)and endoscopic ultrasonography(EUS)can further analyze the characteristics of these tumors, and endoscopic submucosal dissection(ESD)remains the primary treatment modality for early-stage g-NEN.

Since the outbreak of COVID-19, many skin diseases related to or secondary to COVID-19 have been reported globally, and the infection can trigger or exacerbate skin diseases and other systemic symptoms. The review aims to discuss the manifestations and management of severe skin diseases following COVID-19, such as systemic lupus erythematosus, severe psoriasis, pemphigus, dermatomyositis, and Beh?et's syndrome, with the goal of improving the diagnosis and treatment of severe skin diseases after COVID-19.

Objective:To explore the correlation between serum nidogen-2 (NID-2) and thoracic aortic calcification in patients with chronic kidney disease (CKD), and construct a risk prediction model based on NID-2 to evaluate its value in predicting the risk of the severe thoracic aortic calcification and cardiovascular and cerebrovascular events in CKD patients.Methods:It was a prospective cohort study. Patients with CKD at stage 3 to 5D in the Zhongda Hospital Affiliated to Southeast University from January 2022 to January 2023 were enrolled. Syngo.via software was used to evaluate the volume of thoracic aortic calcification, and enzyme-linked immunosorbent assay was employed to determine the level of serum NID-2. According to the volume of thoracic aortic calcification, the patients were divided into three groups: no calcification group, mild calcification group and severe calcification group. The top 25% of the patients were defined as no or mild calcification group, and the latter 75% were defined as severe calcification group. The follow-up period was one year. During the follow-up period, cardiovascular and cerebrovascular events, as well as all-cause death among the enrolled patients were recorded. Logistic regression analysis was used to screen the influencing factors of thoracic aortic calcification. Based on the results of logistic regression analysis, a nomogram prediction model was constructed. The receiver operating characteristic curve (ROC curve), calibration curve, and decision curve were employed to evaluate the discrimination, calibration and clinical practicality of the nomogram model.Results:A total of 132 patients were included, with 91 males (68.94%) and age of (56.51±16.37) years. There were 60 CKD 3-5 stage patients (non-dialysis, 45.45%) and 72 CKD 5D patients (dialysis, 54.55%). Serum ND-2 levels differed significantly among healthy individuals, dialysis patients and non-dialysis patients ( H=70.651, P<0.001). There was no statistically significant difference in serum NID-2 level between the no or mild calcification group and the severe calcification group in dialysis patients ( Z=350.00, P=0.426). The serum NID-2 level in the severe calcification group was significantly higher than that in the no or mild calcification group in non-dialysis patients ( Z=242.00, P=0.019). In non-dialysis patients, there was a statistically significant correlation between serum NID-2 level and volume of thoracic aortic calcification ( r=0.40, P<0.001). In dialysis patients, there was no statistically significant correlation between serum NID-2 level and volume of each segment of thoracic aortic calcification (all P>0.05). The univariate logistic regression analysis showed that, age, hemoglobin, serum albumin, estimated glomerular filtration rate, NID-2, hypertension, type 2 diabetes mellitus and cerebral infarction were correlated factors of thoracic aortic calcification in non-dialysis patients (all P<0.05). Multivariate logistic regression analysis showed that age ( OR=1.22, 95% CI 1.08-1.50, P=0.010) was an independent correlated factor of thoracic aortic calcification in non-dialysis patients. The above related variables of univariate logistic regression analysis were incorporated into a nomogram to construct a predictive model for severe vascular calcification in non-dialysis patients, yielding an AUC of 0.94 (95% CI 0.89-0.99) in ROC curve, with a sensitivity of 83% and a specificity of 95%. A nomogram model based on above variables for predicting cardiovascular and cerebrovascular events in non-dialysis patients demonstrated an AUC of 0.95 (95% CI 0.90-1.00) in ROC curve, with a sensitivity of 95% and a specificity of 87%. Conclusions:In non-dialysis patients, serum NID-2 level in the severe calcification group is significantly higher than that in the no or mild calcification group. The serum NID-2 is a related factor of thoracic aortic calcification and cardiovascular and cerebrovascular events in non-dialysis patients. The nomogram prediction model constructed by combining NID-2 with age, hemoglobin, serum albumin, estimated glomerular filtration rate, hypertension, type 2 diabetes mellitus and cerebral infarction has a high predictive value for the risk of thoracic aortic calcification as well as cardiovascular and cerebrovascular events in non-dialysis patients.

Objective:To investigate the factors affecting the efficacy of 131I treatment for Graves′ disease (GD) and to construct a predictive model for the treatment outcomes of 131I therapy. Methods:Retrospective analysis of the treatment efficacy was performed on 2 190 patients (547 males, 1 643 females, age (42.9±12.4) years) with GD, who received initial 131I treatment in Tianjin Medical University General Hospital between October 2013 and May 2018. Univariate analysis ( χ2 test, et al) and logistic regression were performed to analyze the possible factors affecting the efficacy of 131I treatment. An efficacy prediction model for 131I treatment of GD was constructed, and decision curve analysis (DCA) was used to evaluate the clinical utility of the prediction model. Results:The overall effectiveness rate of 131I treatment for GD patients was 99.95%(2 189/2 190), with a total cure rate of 83.74%(1 834/2 190), among which 94.11%(1 726/1 834) were cured after a single treatment. Pre-treatment thyroid mass was identified as an independent risk factor affecting the efficacy of initial 131I treatment (odds ratio ( OR)=0.983(95% CI: 0.977-0.989), P<0.001). The clinical cure rate was higher in patients who received an adequate dose of 131I compared with that in patients who didn′t receive an adequate dose (79.97%(1 537/1 922) vs 70.52%(189/268); χ2=12.57, P<0.001), but it did not increase the incidence of hypothyroidism within one year. A predictive model was constructed, and it was found that thyroid mass and disease duration had a relatively high impact on the clinical cure rate. The concordance index (C-index) of the predictive model was 0.623(95% CI: 0.593-0.654). DCA indicated that the predictive model offered substantial net benefits across a wide range of probability thresholds. Conclusions:131I treatment is effective in most patients with GD. The predictive model for efficacy of initial 131I treatment developed in this study can assist in evaluating treatment outcomes and help clinicians select the most suitable 131I treatment dose, enhancing clinical decision-making.

Objective To establish a mouse model of bronchiectasis with acute infection and evaluate the immunogenicity and protective effect of a self-assembling Pseudomonas aeruginosa(PA)nanoparticle vaccine rePO-FN based on fusion of PcrV-OprI(rePO)protein with self-assembling ferritin(Ferritin).Methods ① SPF-grade female C57BL/6 mice(aged 6～8 weeks,weighing 18～20 g)were randomly allocated into normal saline group,and low-,medium-and high-dose elastase groups(n=6).A mouse model of bronchiectasis was established via intratracheal instillation of different doses of elastase(30 μL of normal saline containing 0.65,1.30 and 2.60 IU elastase)for 3 consecutive days.At 14 and 21 d after modeling,ELISA and HE staining were performed respectively to detect the concentration of IL-6 and to observe pathological changes in lung tissue in order to confirm the modeling.② A recombinant plasmid encoding the gene of fusion protein rePO-FN was constructed and expressed in E.coli.The target protein was purified via affinity chromatography and renatured to obtain the desired protein.The physicochemical properties of the rePO-FN protein were characterized using SDS-PAGE protein gel electrophoresis,dynamic light scattering,molecular sieve chromatography,and transmission electron microscopy.③ C57BL/6J mice were randomly divided into PBS group,rePO group,rePO-FN group,and Ferritin group(n=10).The mice in the above groups were immunized intramuscularly with 100 μL PBS buffer alone or containing 10 μg of corresponding proteins on days 0,7,and 14.ELISA was used to measure the specific antibodies in serum.In 7 d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the PBS,rePO,and rePO-FN groups.After establishing a bronchiectasis model by intratracheal instillation of 2.60 IU of elastase in C57BL/6J mice as described above,the mice were randomly divided into bronchiectasis PBS group,bronchiectasis rePO group,and bronchiectasis rePO-FN group(n=10).Immunization was conducted at the same dose and procedure as described above,in 21 d after bronchiectasis modeling.At the 7th d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the groups.Results ①Repeated intratracheal instillation of elastase significantly increased the concentration of IL-6 in the lung tissue when compared to the content of the normal saline group(P＜0.05).Pathological observations revealed varying degrees of bronchial wall destruction,alveolar fusion,edema,neutrophil infiltration,and hemorrhage,with the severity increasing with elastase dose,which confirming successful establishment of the mouse model of bronchiectasis.② Well-dispersed rePO-FN nanoparticles were successfully prepared,with an average particle size of 91.28 nm,a Zeta potential of approximately-6.5 mV,and a polydispersity index(PDI)of 0.306.Molecular sieve chromatography determined the elution volume of rePO-FN protein to be 8.80 mL,corresponding to a molecular weight of approximately 1 400 kDa.③ Under acute PA XN-1 strain infection,the survival rate of the rePO-FN immunization group and the bronchiectasis rePO-FN immunization group were significantly higher than that of the PBS control group(P＜0.05).Additionally,bacterial colonization in the lung tissues was significantly lower in the rePO-FN immune group and the bronchiectasis rePO-FN immune group under acute PA XN-1 strain infection than that in the rePO group and the bronchiectasis rePO group(P＜0.05).Conclusion Our vaccine rePO-FN can effectively trigger a strong humoral immune response and provide significant protection against PA infection in a mouse bronchiectasis model.