To establish a mouse model of infection with the minimum lethal dose of Vibrio vulnificus (V. vulnificus) and to evaluate the protective efficacy of inactivated whole-cell (IWC) vaccine using this model.

A mouse model of lethal-dose infection was established by intraperitoneal injection of different doses of V. vulnificus. Bacterial colonization in the organs was detected with tissue homogenate plating, and pathological changes in the organs were observed after tissue section staining. Flow cytometry was used to detect immune cell responses after liver tissues were digested into single-cell suspension. IWC vaccine of V. vulnificus was prepared, and the mice were immunized through different routes to observe the protective efficacy of the vaccine.

A mouse model of infection with the minimum lethal dose at 1×10⁶ CFU of V. vulnificus was successfully established. After infection, the bacteria weremainly colonized in the liver of mice and caused severe pathological damages. Compared with the uninfected mice, the proportion of neutrophils in the liver was significantly increased in the infected mice, whereas the proportions of B cells and T cells were correspondingly decreased (P<0.05). A single intramuscular or intraperitoneal injection of the IWC vaccine could protect the mice effectively against lethal infection of V. vulnificus in 7 d later (P<0.01), although the level of serum IgG having no significant increase.

A mouse model of lethal-dose infection with V. vulnificus is successfully established, with histopathological characteristics. The IWC vaccine of V. vulnificus rapidly mediates immune protection in this model probably independent of IgG.

To develop a composite nutritional preparation that can effectively improve exercise performance under calorie restriction (CR) condition.

A total of 24 male C57BL/6J mice (weighing 23~26 g, 8 weeks old) were randomly divided into control group (CON), CR, CR+basal nutrient group (CRN1), and CR+compound nutrient group (CRN2). All groups underwent moderate-intensity running training 5 d per week, for totally 3 weeks. The grip strength of the forelimbs were measured weekly, and in 3 weeks after training, exhaustion and post-exhaustion distance tests were conducted to evaluate exercise performance. Blood biochemical indicators, levels of skeletal muscle and liver redox biomarkers, and histopathological conditions were measured and observed.

After 21 d of intervention, the CR group and CRN1 group had the post-exhaustion running distance prolonged by 278% and 289%, respectively, reduced blood glucose level, and decreased muscle mass, subcutaneous fat and epididymal fat mass when compared with the CON group (P<0.05). Compared with the CON group, the CRN1 and CRN2 groups demonstrated significantly higher gastrocnemius glycogen content. The CRN2 group obtained even longer post-exhaustion distance (increased by 52% and 36% respectively, compared with the CON group and CRN1 group, P<0.05), enhanced grip strength of the forelimbs (raised by 9%, 17% and 15%, respectively than the CON, CR and CRN1 groups, P<0.05), elevated brown fat mass (compared to the CON group and CRN1 group, P<0.05), increased blood glucose level (compared to the CRN1 group, P<0.05), decreased blood low-density lipoprotein cholesterol level (compared to the CON and CR groups, P<0.05), and increased glutathione peroxidase content in the gastrocnemius muscle (compared to the CON group, P<0.05).

Supplementing with compound nutritional supplements in mice under CR can promote exercise performance, including improving fatigue recovery after exhaustive exercise and enhancing forelimb grip strength.

To explore the role and mechanism of ferroptosis in vitamin D3 (VD3) improving the cytotoxicity of keratinocytes (HaCaT cells) induced by nitrogen mustard (NM).

HaCaT cells were treated with gradient concentrations of NM (5, 10, 20, and 50 μmol/L) alone or in combination with ferroptosis inhibitor ferrostatin-1 or liproxstatin-1 (10 μmol/L), Nrf2-specific agonist tBHQ (10 μmol/L), and VD3 (10 nmol/L), respectively. After co-culture for 24 h, cell viability was assessed, and the intracellular contents of glutathione (GSH) and malondialdehyde (MDA) were measured. Additionally, the expression levels of Nrf2, xCT, and GPX4 were detected.

NM exerted inhibitory effect on the proliferation of HaCaT cells in a dose-dependent manner, and the cell viability was reduced to approximately 55% at an NM concentration of 20 μmol/L (P<0.05). Ferroptosis inhibitors significantly mitigated the cytotoxic damage induced by NM in HaCaT cells (P<0.05), but activation of nuclear factor E2 related factor 2（Nrf2） markedly attenuated NM-induced ferroptosis, as indicated by the restoration of intracellular GSH level and the decrease in MDA content (P<0.05). VD3 specifically targeted the Nrf2 signaling pathway, upregulated the expression of xCT and GPX4 protein, thereby inhibiting ferroptosis and reducing NM-induced cytotoxicity in HaCaT cells.

VD3 mitigates NM-induced cytotoxicity in HaCaT cells by inhibiting ferroptosis via Nrf2 activation.

To determine the effects of hypoxia pre-treatment combined with radiation damage on the hematopoietic cells in the bone marrow of mice.

A total of 165 male C57BL/6 mice (10~12 weeks old, weighing 20~25 g) were randomly divided into 7 groups: normal control (Control, n=33), 6 Gy irradiation (6-Gy, n=43), 7 d hypoxia-6 Gy irradiation (Hy-7 d+6 Gy, n=43), 7 Gy irradiation (7 Gy, n=12), 7 d hypoxia-7 Gy irradiation (Hy-7 d+7 Gy, n=12), 7 Gy continuous hypoxia treatment (Hy-7 d+7 Gy+Hy, n=12), and 6 Gy continuous hypoxia treatment (Hy-7 d+6 Gy+Hy, n=10). The mice of the hypoxia treatment groups were given 7-day hypoxic pretreatment (12% oxygen) in a normobaric hypoxic chamber, while those of the other groups were housed in normoxic condition. After pretreatment, the mice of the irradiation groups were exposed to a single 6 or 7 Gy of whole-body ⁶⁰Co γ-irradiation in normoxia. The mice of the hypoxia and irradiation groups were kept in hypoxic condition in 24 h post-irradiation followed by being resumed to normoxia, while those of the continuous hypoxia treatment groups were remained in hypoxia. After bone marrow cell suspensions were prepared from the Control, 6 Gy, and Hy-7 d+6 Gy groups, bone marrow nucleated cells (BMNCs) were counted via automated cell counter. HE staining was employed to observe pathologic changes in medullary cavity, and flow cytometry was used to assess Lin^-Sca1⁺c-Kit⁺ (LSK) hematopoietic stem/progenitor cells, myeloid progenitors (MPs), and mature T/B/myeloid cells. The mice of the 7 Gy, Hy-7 d+7 Gy, and Hy-7 d+7 Gy+Hy groups were monitored for 30-day survival after hypoxic pretreatment. The dynamic changes in the counts of red blood cells (RBC), white blood cells (WBC) and platelets (PLT), and hemoglobin (HGB) level were observed in the 6 Gy, Hy-7 d+6 Gy, and Hy-7 d+6 Gy+Hy groups with aid of a fully automatic blood analyzer. Single-cell RNA sequencing was performed on bone marrow cell suspension derived from the mice euthanized in 17 d after irradiation from the Control, 6 Gy, and Hy-7 d+6 Gy groups.

①Compared to the Control group, the 6 Gy group showed significantly reduced BMNCs (P<0.01), dilated bone marrow sinusoids, and erythrocyte extravasation. The Hy-7 d+6 Gy group exhibited higher cellular density and attenuated BMNC loss than the 6 Gy group (P<0.01). ②Flow cytometry revealed less LSK, MP, and mature T/B/myeloid cells in the 6 Gy group than the Control group (P<0.05), and the reduced counts of LSK and MP were mitigated in the Hy-7 d+6 Gy group (P<0.01). ③The Hy-7 d+7 Gy group demonstrated improved 30-day survival than the 7 Gy group (P<0.01), while continuous hypoxia (Hy-7 d+7 Gy+Hy) failed to enhance the survival. No statistical difference was seen in the survival rate between the 2 groups (P=0.12), though the Hy-7 d+7 Gy group showing higher survival rate. ④Routine blood test revealed that the Hy-7 d+6 Gy group showed faster WBC recovery (vs the 6 Gy and Hy-7 d+6 Gy+Hy groups, P<0.05), higher pre-irradiation RBC/HGB levels, and accelerated PLT restoration (P<0.05). ⑤Single-cell RNA sequencing indicated that hypoxia pretreatment suppressed the numbers of long-term hematopoietic stem cells/short-term hematopoietic stem cells (LT-HSC/ST-HSC) depletion in the Hy-7 d+6 Gy group when compared with the 6 Gy group, which was consistent with the results of flow cytometry. Pseudotime trajectory aligned the Hy-7 d+6 Gy group, as the Control group, showed enriched undifferentiated LSKs. Differential gene analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that oxidative phosphorylation pathway was strongly activated in the 6 Gy group, while the Hy-7 d+6 Gy group had enriched in chromatin remodeling and mRNA surveillance pathways.

Hypoxic preconditioning alleviates radiation-induced bone marrow injury, and post-irradiation normoxia restoration promotes hematopoietic recovery in acute radiation-exposed mice.

To investigate the role and possible mechanism of celastrol (Cel) in sodium oxalate (NaOx)-induced acute kidney injury (AKI) and crystal deposition in the kidney tissues in mice.

Male C57BL/6 mice (aged 8~12 weeks, weighing 22~24 g) were randomly divided into 3 groups. Saline group (control group, intraperitoneal injection with normal saline and drinking water freely), NaOx group (injured group, intraperitoneal injection of 75 mg/kg NaOx, and drinking water containing 50 μmol/L NaOx), and NaOx+Cel group (treatment group, intraperitoneal injection of 1 mg/kg Cel firstly and then 75 mg/kg NaOx in 24 h later, drinking water containing 50 μmol/L NaOx). All specimens were collected in 24 h after NaOx injection. HK-2 cells were randomly divided into 4 groups: Medium group (no treatment), NaOx group (500 μmol/L NaOx), NaOx+Cel group (400 nmol/L Cel pre-treatment for 2 h followed by 500 μmol/L NaOx treatment), and NaOx+Cel+BA group [8 μmol/L betulinic acid (BA, NF-κB agonist) after the interventions as the NaOx+Cel group]. Cells of each group were collected in 24 h after corresponding treatments. Von Koosa and cell adhesion assays were used to observe crystal deposition. HE staining was employed to observe renal histopathology and score the damage. CCK-8 assay was utilized to detect cell viability to obtain the optimal concentrations of NaOx and Cel. Serum urea and creatinine levels were detected. Immunohisotochemical assay was conducted to detect the expression of OPN, CD44, KIM-1, NGAL, p65, IL-1β, BAX, and Caspase-3, and Western blotting was performed for protein levels of OPN, CD44, KIM-1, p65, P-p65 and IL-1β.

The mice in the NaOx+Cel group showed reduced crystal deposition (P<0.0001), attenuated renal tubular damage (P<0.01), decreased serum urea and creatinine levels (P<0.05), and declined expression levels of the renal adhesion molecules OPN and CD44, the kidney injury molecules KIM-1 and NGAL, the inflammation-associated molecules p65 and IL-1β, and the apoptosis related molecules BAX and Caspase-3 when compared with the NaOx group (P<0.05). In in vitro study, the NaOx+Cel group showed reduced crystal adhesion (P<0.0001), decreased expression of the adhesion molecules OPN and CD44 (P<0.05), down-regulation of the inflammatory molecule IL-1β and P-p65/p65 ratio (P<0.05), and down-regulation of the renal injury molecule KIM-1 (P<0.05) when compared with the NaOx group. In the NaOx+Cel+BA group, crystal adhesion was significantly increased (P<0.0001), the inflammatory molecule IL-1β and the ratio of P-p65/p65 were increased (P<0.05), and the kidney injury molecule KIM-1 was increased when compared with the NaOx+Cel group (P<0.05).

Cel may reduce NaOx-induced crystal deposition and AKI by inhibiting NF-κB activation.

To investigate the differential expression genes (DEGs) related to angiogenesis in rosacea (RA) by utilizing bioinformatics analysis in order to screen the key genes and verify their mRNA expression levels.

The gene microarray dataset GSE65914 was retrieved from the Gene Expression Omnibus (GEO) repository. Analyzed by R programming, the dataset was refined to identify DEGs related to RA, and then cross-referenced with angiogenesis-related genes from the GeneCards database to get a subset specific to RA angiogenesis. The process of identifying key genes was augmented by employing protein-protein interaction (PPI) network analysis and Cytoscape-based computational algorithms. The mRNA expression levels of the aforementioned pivotal genes were detected by real-time fluorescent quantitative reverse transcription PCR (RT-qPCR).

A total of 947 RA-associated DEGs were identified from GEO dataset, and then 202 genes related to RA angiogenesis were further delineated. PPI network analysis and Cytoscape algorithm finally identified 3 key genes, that is, CXCL8, IL-1B, and STAT1. The results of RT-qPCR showed that the mRNA expression levels of MIP-2, GCP-2, IL-1B and STAT1 in RA lesions were significantly higher than those in normal controls (P<0.05).

With aid of bioinformatics analysis, our study has screened and validated key genes associated with angiogenesis in RA, namely CXCL8, IL-1B, and STAT1, which providing a theoretical basis for elucidating the potential mechanisms underlying RA-induced angiogenesis and developing targeted therapeutic strategies.

To construct a preoperative prediction model for proliferative hepatocellular carcinoma(HCC) based on enhanced CT image features, and to analyze the prognosis of the disease.

A retrospective case-control study was conducted on 603 patients with pathologically confirmed HCC. Among them, 519 cases from the First Affiliated Hospital of Army Medical University were randomly divided into a training group(n=363) and an internal verification group(n=156) in a ratio of 7:3. Another 84 patients from the Second Affiliated Hospital of Chongqing Medical University served as an external validation group. All patients underwent abdominal CT scan with contrast before surgery. The clinical data and CT imaging characteristics of proliferative and non-proliferative HCC patients were observed. Binary logistic regression analysis was used to identify the independent risk factors of proliferative HCC, and a nomogram prediction model was constructed. Receiver operating characteristic(ROC) curve was plotted to evaluate its diagnostic performance, and calibration curve and decision curve analysis(DCA) were applied to evaluate its calibration performance and clinical application value. The model was validated in both the internal and external validation groups. Kaplan-Meier survival curves were employed to compare the prognosis between proliferative and non-proliferative HCC.

Multivariate analysis showed that negative result of HBV-DNA quantification, incomplete tumor capsule, intratumoral necrosis or ischemia(≥20%), and annular hyperenhancement in arterial phase were independent predictors in predicting proliferative HCC(P<0.05). Our nomogram model for predicting proliferative HCC based on the above clinical imaging features had an AUC value of 0.805(95%CI: 0.756~0.854), a sensitivity of 83.19% and a specificity of 64.80% in the training group. For the internal validation group, the AUC value was 0.793(95%CI: 0.721~0.854), the sensitivity was 67.86%, and the specificity was 75.00%. In the external validation group, the AUC value was 0.842(95%CI: 0.746~0.912), the sensitivity was 72.41%, and the specificity was 90.91%. Calibration curve and DCA showed that the model had good calibration performance and clinical applicability. The disease free survival(DFS) of the patients with proliferative HCC diagnosed by pathologically confirmed/predictive models was significantly shorter than that of non-proliferative HCC patients(training group: P=0.005, P<0.001; internal validation group: P=0.006, P=0.006; external validation group: P=0.002, P=0.015).

Our prediction model based on clinical and imaging features can accurately diagnose proliferative HCC before surgery, and the prognosis of proliferative HCC is generally poor.

To explore whether the biopsy rate and the positive rate of biopsy during colonoscopy can serve as effective indicators for evaluating the diagnostic proficiency of endoscopists.

Total fifteen endoscopists from the First Affiliated Hospital of Army Military Medical University, Guiqian International General Hospital and No. 958 Hospital of PLA Army were enrolled and served as the study subjects. According to their years of experience, gender and working hours, they were divided into a senior group (n=8) and a junior group (n=7), a male group (n=9) and a female group (n=6), and a morning period (08:00-12:00) and an afternoon period (13:00-18:00). The data of all patients examined by the aforementioned endoscopists between January 2023 and December 2023 were collected, including the basic demographics, colonoscopic results, biopsy outcomes and pathological diagnoses of the patients. ANOVA was used to analyze the differences in the biopsy rate and positive rate of biopsy existed among the endoscopists of different years of experience, gender and working at different periods (morning and afternoon). Additionally, multivariate logistic regression analysis was employed to identify the independent risk factors associated with positive rates of biopsy.

The senior group had significantly lower biopsy rate [(5.73±0.81)% vs (6.89±0.97)%], but obviously higher positive rate of biopsy [(53.52±3.87)% vs (33.70±7.85)%] than the junior group (both P<0.01). The biopsy rate was notably higher during the morning working time period than the afternoon working time period in all the endoscopits [(6.76±1.11)% vs (5.53±1.57)%, P<0.05]. Logistic regression analysis showed that the years of experience [(53.52±3.87)% vs (33.70±7.85)%, OR=92.187, 95%CI: 3.118~2 725.366, P<0.01) and working time [(48.35±10.50)% vs (37.80±10.65)%, OR=20.885, 95%CI: 1.266~344.602, P<0.05) were influencing factors for positive rate of biopsy.

The biopsy rate is low and positive rate of biopsy is high among the senior experienced endoscopists, while, opposite results are seen in the junior experienced clinicians, indicating correlation of biopsy rate and positive rate of biopsy with diagnostic proficiency of endoscopists.

To prepare an advanced irrigation device equipped with a specialized nozzle designed to access the pericoronal blind sac of impacted wisdom teeth, and evaluate its efficacy in plaque removal.

3D-printer was utilized to develop pericoronal models simulating mandibular wisdom teeth with high vertical impaction and varying soft tissue coverage (0%, 25%, 50%, and 75%, respectively) and personalized pericoronal irrigator. The obtained pericoronal models were randomly divided into 3 groups, treated with personalized precision pericoronal flushing, routine flushing and brushing, respectively, and, under a pressures of 45 psi or 75 psi （1 psi=6 894.76 Pa）, were performed to clean the test surfaces of plaque-biofilm-obstructed wisdom teeth, respectively. Images of the tooth surfaces were analyzed before and after cleaning, and the cleaning rate was calculated using Python software.

For wisdom teeth with 50% and 75% pericoronal soft tissue coverage, at a pressure of 45 psi, the precision pericoronal flushing group demonstrated significantly higher cleaning rate when compared to the brushing and routine flushing groups (P<0.05). At 0% and 25% pericoronal soft tissue coverage, the precision pericoronal flushing and brushing groups exhibited significantly higher cleaning rates than the routine flushing group (P<0.05). Furthermore, at 75 psi, the precision pericoronal flushing group achieved a significantly higher cleaning rate across all 4 levels of pericoronal soft tissue coverage than both the routine flushing and brushing groups (P<0.05). Notably, the precision pericoronal flushing group at 75 psi obtained significantly higher cleaning rate than at 45 psi across 0%, 25%, and 50% pericoronal soft tissue coverage (P<0.05).

The precision pericoronal irrigation device demonstrates significantly superior cleaning efficacy for vertically impacted mandibular third molars compared to conventional irrigation devices and manual tooth brushing. It is an effective device for preventing and treating pericoronitis.