The pathogenic characteristics of Escherichia coli (E.coli) in bacterial infections were analyzed using a combination of multiple molecular typing techniques in order to provide evidence for the management of clinical medication safety.

Samples from some bacterial infection-related cases in a district of Chongqing in 2021 were collected. A total of 30 E.coli strains were selected by a completely random method, and phoA gene PCR assay was performed for identification. Molecular typing of the strains was analyzed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Antimicrobial susceptibility testing was conducted to determine the drug resistance of the strains, and four β-lactamase-encoding genes (blaCTX-M, blaTEM, blaSHV, blaZ) were selected to detect the carriage of resistance genes.

All 30 E.coli strains displayed the phoA gene target band. Their PFGE banding patterns, with a similarity of 50%~98%, could be classified into 8 clusters. Cluster C was the dominant group, accounting for 53.3% (16/30). C1 and C2 exhibited high genetic correlation, indicating a close phylogenetic relationship. One E.coli strain could not be assigned a sequence type (ST) by MLST, while the remaining 29 E.coli isolates were classified into 16 different STs, demonstrating a polymorphic distribution. Among them, 10 isolates belonged to ST131 (10/30, 33.33%). Evolutionary analysis of the 10 ST131 E.coli strains revealed their distribution across different branches, indicating varying degrees of genetic relatedness. Antimicrobial susceptibility testing revealed that all 30 E.coli strains exhibited varying degrees of resistance, with the highest resistance rate observed against the β-lactam antibiotic ampicillin (25/30,83.33%). Among them, 60.0% were multidrug-resistant bacteria (MDRB). These MDRB strains exhibited 16 distinct resistance profiles, displaying a scattered distribution without a dominant resistance pattern. 50.0% (9/18) of the MDRB strains exhibited six-drug resistance, while the most drug-resistant strain showed eight-drug resistance. Furthermore, the blaCTX-M gene carriage rate among the 30 E.coli strains was 86.67% (26/30), while no blaZ gene was detected.

E.coli related to bacterial infections from a Chongqing district exhibited diverse PFGE/MLST patterns and significant drug resistance. The application of multiple molecular typing techniques can reveal the genetic diversity, evolutionary relationships, and antimicrobial resistance characteristics of pathogenic bacteria. Countermeasures　It is recommended to enhance the molecular typing and drug resistance surveillance network for pathogenic bacteria, establish an early warning mechanism, and implement hierarchical management of antibiotics, thereby improving targeted prevention and epidemic traceability capabilities for key drug-resistant bacteria such as ST131.

To explore the mediating role of mindful attention and awareness in depressive symptoms and insomnia severity among patients with comorbid depression and insomnia.

A cross-sectional study was conducted, enrolling 267 patients with comorbid depression and insomnia who were treated in the outpatient Department of Medical Psychology of Second Affiliated Hospital of Army Medical University, from March to May 2024. Basic demographic and clinical data were collected using a general information questionnaire. Depressive symptom severity was measured via the Patient Health Questionnaire-9 (PHQ-9), insomnia severity via the Insomnia Severity Index (ISI), perceived stress via the Perceived Stress Scale-10 (PSS-10), and mindful attention and awareness via the Mindful Attention Awareness Scale (MAAS). Pearson correlation analysis was used to examine the correlations between depressive severity, insomnia severity, perceived stress, and mindful attention and awareness. Mediation analysis was performed using Process 4.1.

The PHQ-9 score was (13.80±5.98) and the ISI score was (17.10±5.56) in the 267 patients. Pearson correlation analysis showed that depressive severity and insomnia severity were positively correlated with perceived stress (r=0.531, 0.351, P<0.001) and negatively correlated with mindful attention and awareness (r=-0.373, -0.350, P<0.001). Mediation analysis using Process 4.1 indicated that the combined mediating effect of mindful attention and awareness and insomnia between perceived stress level and depressive level was 0.157, with a 95% confidence interval (CI) of 0.102~0.217, and the total mediating effect was significant (P<0.001).

Perceived stress directly positively affects depression and indirectly exacerbates depression through insomnia as a mediator, and mindful attention and awareness can weaken the promoting effect of perceived stress on insomnia.

To determine the expression level of TG-interacting factor 1 (TGIF1) in gastric cancer (GC) and its correlation with prognosis, and investigate the characteristics of immune microenvironment of TGIF1-overexpressing GCs and its clinical significance.

Integrated analysis was performed using transcriptomic data from The Cancer Genome Atlas (TCGA), Asian Cancer Research Group (ACRG), and our in-house GC transcriptome database. Immunohistochemistry (IHC) assay was employed to compare TGIF1 expression between GC and normal gastric mucosa tissues. Based on Kaplan-Meier Plotter online database, the correlation between TGIF1 expression and clinical prognosis was evaluated. Transcriptomic data were analyzed to identify functional enrichment features of GC with high TGIF1 expression. The GENIE3 toolkit and STRING database were utilized to predict TGIF1-regulated target genes and protein-protein interaction (PPI) networks, respectively to explore the potential immune-related signaling pathways regulated by TGIF1. Single-cell RNA sequencing (scRNA-seq) was applied to analyze the association between TGIF1 expression and tumor microenvironment (TME) disorder.

Transcriptomic analysis revealed significantly higher TGIF1 expression in GC tissues compared to normal tissues (P<0.01). Patients with high TGIF1 expression exhibited poorer clinical prognosis (P<0.05). IHC assay confirmed elevated TGIF1 expression in GC tissues than normal tissues (P<0.01). GC with high TGIF1 expression was enriched in immune regulatory pathways, including immunosuppressive cytokines such as chemokine C-C motif ligand 20 (CCL20). These tumors displayed an immunosuppressive TME, characterized by abundant immunosuppressive NK cells, mast cells, regulatory T cells (Tregs), myeloid cells, and exhausted CD8⁺ T cells. Cell interaction analysis suggested that TGIF1-overexpressing GC cells may engage with Tregs via the CCL20-CCR6 axis. Co-high expression of signature gene sets from these interacting cells was associated with significantly shorter survival in the patients (P<0.05).

TGIF1 is highly expressed in GC tissues and may serve as a biomarker for immunosuppressive TME and poor prognosis. TGIF1-overexpressing GC cells may interact with multiple immune cells, particularly Tregs, to induce an immunosuppressive microenvironment.

To elucidate the effects of miR-616 on the malignant biological processes of osteosarcoma and to preliminarily explore its potential mechanisms.

In situ hybridization (ISH) was employed to analyze miR-616 expression in 11 paraffin-embedded osteosarcoma specimens collected in our department during January 2018 to December 2019. Quantitative real-time PCR (qRT-PCR) was used to compare the mRNA expression level of miR-616 in the osteoblast cell line hFOB1.19 and osteosarcoma cell lines 143B and HOS. Stable cell lines with miR-616 knockdown or overexpression were established via lentiviral transfection in 143B and HOS cells. Cell proliferation and apoptosis were detected by flow cytometry, while cell invasion and migration were assessed using Transwell and colony formation assays, respectively. To evaluate the effect of miR-616 on tumor growth in vivo, 10 female nude mice (4 weeks old, weighing 18~20 g) were randomized into a control group and a miR-616 overexpression group. After the xenograft tumor model was constructed, the growth of subcutaneous tumors was monitored. Finally, next-generation sequencing and a dual-luciferase reporter assay were performed to identify the target genes of miR-616.

ISH results showed that miR-616 expression was up-regulated in osteosarcoma tissues than adjacent tissues, and primarily localized in the cytoplasm. qRT-PCR confirmed that miR-616 level was significantly higher in 143B and HOS cells than hFOB1.19 cells (P<0.05). In vitro experiments revealed that miR-616 overexpression enhanced the proliferation, migration and invasion, while suppressing apoptosis in 143B and HOS cells (P<0.01). Conversely, miR-616 knockdown weakened these malignant phenotypes (P<0.05), with miR-616-3p showing a stronger effect on apoptosis than miR-616-5p. Animal experiments demonstrated that the tumor weight in the miR-616 overexpression group was significantly greater than that of the control group (98.00±17.22 vs 33.60±8.08 mg, P<0.01). Furthermore, KLF2 was identified and confirmed as a direct target of miR-616.

MiR-616 promotes malignant biological behaviors in osteosarcoma, and its expression level indicates that it may serve as a potential therapeutic target.

To investigate whether microRNA-21-5p (miR-21) plays a protective role in hyperoxia-induced acute lung injury (HALI) by regulating ferroptosis through the STAT3/P53/SLC7A11 axis.

The interaction between STAT3 and P53 was analyzed by co-immunoprecipitation (Co-IP). Fifty 9-week-old male C57BL/6J mice were randomly divided into normoxia (Control) group, HALI group, miR-21 overexpression (miR-21) group, STAT3 inhibitor (HY-13818) group, and ferrostatin-1 (Fer-1) group. After the mice from the miR-21 group received miR-21-5p AAV6 or empty vector via tracheal catheter instillation, the animals were then monitored for 3 weeks. The HY-13818 group was intraperitoneally injected with HY-13818 (10 mg/kg) 3 times weekly for 2 weeks. The Fer-1 group received 0.8 mg/kg ferrostatin-1 via tail vein injection once daily for 2 consecutive days during modeling. The HALI model was established by exposure to >90% oxygen. After 48 h of hyperoxia, blood samples were collected via orbital sampling for RT-PCR analysis of miR-21 expression. Lung tissues were harvested for wet/dry weight ratio and assessment of histopathological changes via HE staining for lung injury score. Activity of superoxide dismutase (SOD) and contents of malondialdehyde (MDA), Fe²⁺, glutathione (GSH), and reactive oxygen species (ROS) were measured using photocolorimetry, spectrophotometry and fluorometry. Western blotting was used to evaluate the protein expression of STAT3, P53, SLC7A11, and GPX4.

The results of Co-IP showed that STAT3 could bind to P53. The HALI group exhibited obviously destroyed alveolar structure, disordered arrangement, thickened interval, with a large number of infiltrated neutrophils and collapsed alveoli, and had significantly increased pathological score of lung injury and ratio of lung W_wet/D_dry weight when compared with the Control group (P<0.05). In the miR-21 group, HY-13818 group and Fer-1 group, the severity of lung injury was significantly reduced, and the pathological score of lung injury and the ratio of W_wet/D_dry weight were decreased (P<0.05) when compared with the HALI group. Compared with the control group, the contents of MDA, Fe²⁺ and ROS were increased (P<0.05), the activity of SOD and content of GSH were declined (P<0.05), the protein levels of STAT3 and P53 were increased (P<0.05), and those of SLC7A11 and GPX4 were decreased (P<0.05) in the HALI group. Compared with the HALI group, decreased MDA and ROS levels (P<0.05), enhanced SOD activity, Fe²⁺ and GSH levels (P<0.05), down-regulation of STAT3 and P53 (P<0.05) and up-regulation of SLC7A11 and GPX4 (P<0.05) were observed in the miR-21 group and HY-13818 group.

MiR-21 alleviates HALI, which may be related to its inhibition of ferroptosis through the STAT3/P53/SLC7A11 axis.

To investigate the effects of paeoniflorin (PF) on lipopolysaccharide (LPS)-induced aerobic glycolysis in renal tubular epithelial cell line HK-2 and its underlying mechanism of action.

This study consists of a preliminary experiment and a formal experiment. Preliminary experiment: CCK-8 assay and RT-qPCR were used respectively to measure cell viability and mRNA expression levels of inflammatory factors in HK-2 cells after LPS stimulation to determine the optimal LPS concentration for modeling as well as to evaluate the toxicity of PF and screen for its appropriate concentration. Formal experiment: HK-2 cells were divided into control group (CON group), LPS group, LPS+PF group and LPS+PF+740Y-P group. LPS was used to establish a cell model of sepsis associated-acute kidney injury (SA-AKI) in HK-2 cells, and then the cell model was treated with PF and PI3K activator 740Y-P, correspondingly for 24 h. CCK-8 assay was employed to detect cell viability, and Extracellular Acidification Rate (ECAR) Kit was utilized to measure the rate. The contents of IL-1β, IL-18, lactic acid (Lac) and lactate dehydrogenase A (LDHA) were determined with ELISA. Western blotting was applied to detect the expression of p-PI3K, p-AKT, HIF-1α, pyruvate kinase 2 (PKM2, a key enzyme in aerobic glycolysis) and NOD-like receptor thermal protein domain associated protein 3 (NLRP3), and immunofluorescence assay was performed to observe the expression and distribution of PKM2.

① Our preliminary experiment identified that the optimal concentration of LPS for modeling was 20.0 μg/mL, a safe dosage range of PF was 0~100.0 μmol/L, and its optimal therapeutic concentration was 25.0 μmol/L. ② Compared with the CON group, LPS stimulation resulted in significantly decreased cell viability (P<0.05), increased ECAR (P<0.05), elevated contents of IL-1β, IL-18, Lac and LDHA (P<0.05), up-regulated protein levels of p-PI3K, p-AKT, HIF-1α, p-PKM2 and NLRP3 (P<0.05), and enhanced fluorescence intensity of PKM2 in the nucleus of cells (P<0.05). Compared with the model group, PF treatment reversed all above effects induced by LPS stimulation (all P<0.05). Compared with the LPS+PF group, in the LPS+PF+740Y-P group, ECAR was elevated (P<0.05), the contents of IL-1β, IL-18, Lac and LDHA were increased (P<0.05), and the relative expression levels of p-PI3K, p-AKT, HIF-1α, p-PKM2 and NLRP3 were increased (P<0.05), and the fluorescence intensity of PKM2 was strengthened (P<0.05) and enhanced in the nucleus (P<0.05).

PF reduces aerobic glycolysis in HK-2 cells and alleviates the inflammatory response by inhibiting the PI3K/AKT/HIF-1α signaling pathway.

To investigate the protective effects of berberine (BBR) against drug-induced liver injury (DILI) in mice and the change profiles in gut microbiota, and to elucidate the underlying mechanism by which BBR alleviates DILI through modulation of the gut microbiota.

Acetaminophen (APAP) administration was performed on C57BL/6J mice with or without pre-treatment of ampicillin-neomycin-metronidazole-vancomycin (ABX, depletion of gut microbiota) to establish a DILI model. Based on body weight, 12 mice were randomly divided into an APAP group and a BBR group, with 6 animals in per group. Serum and liver tissue samples were collected from both groups to determine the effects of intragastrical administration of BBR on serum transaminase levels, inflammatory indicators, and liver histopathology. Fresh fecal samples were also collected to profile the gut microbiota using 16S rRNA gene sequencing. Then the alpha and beta diversity, as well as the overall microbial community structure, were compared between the 2 groups. Pearson correlation analysis was employed to analyze the relationships between differentially abundant bacteria and the serological indicators.

Intragastrical administration of BBR significantly reduced APAP-induced serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and ameliorated the proportion of hepatocellular necrosis caused by APAP. The results of 16S rRNA gene sequencing revealed that there were no statistical differences in the alpha diversity indices (Chao1 and Shannon) between the APAP-BBR and APAP groups, but a significant difference was observed in beta diversity (P<0.05). Compared to the APAP-BBR group, the relative abundances of Alloprevotella and Ruminococcaceae_UCG_014 were significantly increased in the APAP group (P<0.05). Conversely, the relative abundance of Parabacteroides was significantly higher in the APAP-BBR group than the APAP group (P<0.05). Pearson correlation analysis demonstrated that Alloprevotella was strongly correlated with AST level (P<0.05), while both Ruminococcaceae and Parabacteroides showed strong correlations with ALT and AST levels (P<0.05).

Intragastric administration of BBR can mitigate the progression of DILI in mice, potentially through modulating the gut microbiota.

To investigate the influencing factors for stroke-associated pneumonia (SAP) in acute ischemic stroke (AIS) patients after endovascular treatment (EVT).

A retrospective case-control trial was conducted on 426 AIS patients with large vessel occlusion (LVO) in anterior circulation admitted in the neurological departments from First Affiliated Hospital of Army Medical University and Zigong Third People’s Hospital during January 2017 and April 2021. Based on SAP occurrence or not, they were divided into an SAP group and a non-SAP group. Demographic information (gender and age), TOAST stroke subtypes (large artery atherosclerosis type, cardiac embolism type, others), vascular risk factors (hypertension, hyperlipidemia, diabetes, atrial fibrillation, smoking, prior stroke history, smoking), and post-onset clinical data [dysphagia, LDL cholesterol, white blood cells, neutrophils, baseline and postoperative NIHSS scores, endovascular outcomes (mTICI grade 2b or 3), 90-day good prognosis (mRS 0-1)] were collected and compared between the 2 groups. Multivariate logistic regression analysis was performed using the parameters with P<0.1 in univariate analysis as independent variables to investigate factors influencing SAP occurrence after EVT in AIS patients.

Among the 426 participants, SAP occurred in 194 cases (45.5%). Multivariate logistic regression analysis revealed that admission white blood cell count (OR=1.125, 95% CI: 1.043~1.213, P=0.000 2), postoperative NIHSS score (OR=1.019, 95% CI:1.001~1.037, P=0.041), and male (OR=1.687, 95% CI:1.078~2.638, P=0.022) were associated with SAP occurrence after EVT in AIS patients.

Higher admission white blood cell count, elevated postoperative NIHSS score, and male gender are risk factors for SAP in AIS patients after EVT. These risk factors should be focused on clinical practice to control SAP incidence.

To quantitatively evaluate age-, sex-, and segment-related characteristics of lumbar disc degeneration (LDD) in patients with low back pain (LBP) using T1rho MRI, and assess its diagnostic value in early-stage disc degeneration.

A retrospective cross-sectional study was conducted on 89 LBP patients (balanced distribution across ages 20~29, 30~39, 40~49, 50~59 years and genders) admitted in Department of Orthopedics of First Affiliated Hospital of Army Medical University from March 2024 to February 2025. All patients underwent lumbar MRI including T1rho sequences. Two senior radiologists performed blinded Pfirrmann grading on 445 intervertebral discs and measured T1rho values of the nucleus pulposus (NP). Kappa test was used to evaluate the inter-observer agreement of Pfirrmann grading, and Spearman correlation coefficient was employed to analyze the correlation between Pfirrmann grade and NP-T1rho value. Disc degeneration characteristics were compared across different ages, sexes, and spinal segments. After adjusting for body mass index (BMI) and gender, partial correlation analysis was used to assess the correlation between NP-T1rho value of each lumbar segment and age. Receiver operating characteristic (ROC) curve was plotted to analyze the diagnostic performance of T1rho quantification for early LDD in LBP patients.

Good inter-observer agreement was observed in Pfirrmann grading across segments, with a Kappa coefficient of >0.70. The NP-T1rho values of all NP were in a strong negative correlation with Pfirrmann grades (r=-0.877, P<0.001). The NP-T1rho values were progressively decreased from Pfirrmann gradesⅠ to Ⅳ (P<0.001), with a steep decline between gradesⅡ and Ⅲ. The patients aged 20~29 and 30~39 years showed significantly higher NP-T1rho values in Pfirrmann grades Ⅰ and Ⅱ discs when compared with those aged 40~49 and 50~59 years (P<0.05). The males had significantly higher mean NP-T1rho values in grade I than the females (P=0.006). In the upper spinal segments (L1~L4), the males exhibited significantly higher mean NP-T1rho value than the females (P=0.025), and NP-T1rho value was decreased obviously with increase of age across all age groups (P<0.05). Except L5/S1 segments, NP-T1rho value and age in all spinal segments had significant negative correlations (r=-0.548~-0.349, P<0.001). The area under the curve (AUC) for distinguishing grade Ⅰ from Ⅱ discs was 0.824 (95%CI: 0.772~0.876), and for grades Ⅱ from Ⅲ was 0.978 (95%CI: 0.964~0.992).

T1rho MRI provides accurate quantitative assessment of early lumbar disc degeneration in LBP patients, with an NP-T1rho value of 79.87 ms serving as a discriminative threshold between grade Ⅱ and grade Ⅲ.