To investigate the interaction between a novel antimicrobial compound, HL-J6, and penicillin-binding protein 1 (PBP1) of Staphylococcus aureus.

With MRSA252 genomic DNA as the template and PBP1F and PBP1R as primers, the expression plasmid pET30a-pbp1-39-608 was constructed by amplifying the target gene fragment followed by cloning into the Nde I/Xho I restriction sites of the pET30a vector. Then the obtained plasmids were transformed into Escherichia coli for the expression of PBP1-39-608 protein, and the product was purified by affinity chromatography. The inhibitory effect of HL-J6 on the transpeptidase activity of PBP1-39-608 was measured using peptidoglycan side chain backbone peptide, with thiol ester analog S2d as the substrate. The affinity between HL-J6 and PBP1-39-608 was detected using microscale thermophoresis (MST), and the binding interaction was confirmed by cellular thermal shift assay (CETSA). Molecular docking and dynamics simulation were performed using AutoDock Vina and Desmond software, respectively, to elucidate the binding mode of HL-J6 with the PBP1-39-608 protein and the key amino acid residues involved.

The recombinant plasmid pET30a-pbp1-39-608 was successfully constructed, and PBP1-39-608 protein was produced after induction and purified, yielding a protein with an approximate molecular mass of 65×10³. HL-J6 inhibited the transpeptidase activity of PBP1-39-608 in a time-dependent manner (P<0.001). The dissociation constant Kd of the binding between HL-J6 and PBP1-39-608 was 64.92 µmol/L. Molecular docking results showed that HL-J6 bound to the active pocket of PBP1-39-608 by interacting with key residues such as ILE-348, ASN-370, THR-516 and PHE-423, with a binding score of -8.38 kcal/mol (<-5.00 kcal/mol). Dynamics simulation results indicated that the complex became stable after 50 ns.

HL-J6 effectively inhibits the transpeptidase activity of Staphylococcus aureus PBP1, and shows stable interaction with the protein.

To explore the efficacy and action mechanism of a novel phenothiazine derivative, DA414, in rats with intracerebral hemorrhage (ICH).

Male Sprague-Dawley (SD) rats (aged 8~10 weeks, weighing 250~300 g) were randomly divided into sham operation, model, and low and high dose DA414 groups [5 and 10 mg/(kg·d)], with20 animals per group. The size and extent of injury in the ICH area were measured by magnetic resonance imaging (MRI) and histopathological slices. Neurological function was evaluated with a behavioral grading system. Western blotting was used to detect the expression of ferroptosis-related factor, glutathione peroxidase 4 (GPX4). Multiplex immunohistochemistry was employed to quantitatively evaluate microglial activation in perihematomal tissue. RT-qPCR was applied to measure the mRNA expression levels of NLRP3 inflammasome components (NLRP3, Caspase-1, IL-1β), pro-inflammatory (IL-18, TLR4, IL-6, TNF-α) and anti-inflammatory cytokines (IL-4, IL-10). The integrity of the blood-brain barrier (BBB) was assessed by Evans blue staining, and the biosafety of DA414 for the liver, kidneys and heart was assessed by HE staining and clinical biochemical tests.

DA414 significantly promoted the absorption of hematoma, reduced neuronal injury, and improved neurological function scores. DA414 significantly up-regulated the ferroptosis regulatory factor GPX4 (P<0.01), and also significantly inhibited the activation of microglia after ICH (P<0.001). RT-qPCR indicated that DA414 treatment resulted in down-regulation of mRNA expression in the inflammasome pathway (NLRP3, Caspase-1, IL-1β, all P<0.01) and pro-inflammatory cytokines (TLR4, IL-6, both P<0.05; IL-18, TNF-α, both P<0.01) and up-regulation of anti-inflammatory cytokines (IL-4, IL-10, both P<0.05), suggesting that DA414 exerts its neuroprotective effect probably by regulating ferroptosis and inflammation. Safety assessment revealed that DA414 had no significant effect on BBB integrity or damage to the liver, kidneys, and heart in rats.

DA414 exerts significant neuroprotective effects in ICH model by targeted inhibition for ferroptosis and modulating inflammatory response. Our study provides an experimental foundation for ICH treatment.

To observe the expression levels of base excision repair (BER) pathway-related proteins in small cell lung cancer (SCLC) tissues, and analyze their relationship with the prognosis and tumor immune microenvironment.

A retrospective cohort study was conducted on 74 patients with limited-stage SCLC undergoing surgical treatment in our medical center from December 2018 to June 2023. Immunohistochemical staining was performed to analyze the protein expression of BER pathway components, apurinic/apyrimidinic endonuclease 1 (APE1), 8-oxoguanine DNA glycosylase 1 (OGG1), DNA polymerase β (POLβ), X-ray repair cross-complementing protein 1 (XRCC1), ATP-dependent DNA ligase I (LIGⅠ), and immune cell infiltration markers of CD3⁺ T cells, CD8⁺ T cells, CD68⁺ macrophages in SCLC tissues. Chi-square test was applied to analyze the relationship of BER protein expression and clinicopathological features; Kaplan-Meier survival curve was plotted to evaluate the impacts of BER protein expression and immune cells on disease-free survival (DFS) and overall survival (OS), multivariate Cox regression analysis was utilized to identify DFS prognostic factors, and Spearman correlation analysis was performed to analyze the correlation of BER-immune cell infiltration. In in vitro experiments, transient transfection was applied in H196 cells to overexpress APE1/POLβ/LIGⅠ, respectively. Thus, the cells were divided into negative control (NC, empty vector) and overexpression (OE, target plasmids) groups. CCK-8 and TUNEL assays were employed to determine the effects of OEAPE1, OEPOLβ and OELIGⅠon cell sensitivity to cisplatin. In in vivo experiments, nude mice bearing xenograft tumors were grouped into WT, E3330 (APE1 inhibitor), cisplatin, and cisplatin+E3330 groups to determine the effects of the combination therapy on tumor growth.

There were no significant correlations of the expression levels of key BER pathway proteins with clinicopathological characteristics, including gender, age, smoking history, tumor location, Ki67 index, or TNM stage (all P>0.05). The patients with low expression of APE1, POLβ, and LIGⅠ had obviously higher DFS rates than those with high expression (P<0.05), and the patients with larger proportion of CD3⁺ T cells also had higher DFS rates than those with smaller proportion (P=0.043). Multivariate Cox regression analysis indicated that tumor TNM stage (HR=2.465) and APE1 expression (HR=2.730) were independent risk factors for the prognosis of SCLC patients (P<0.05). Spearman correlation analysis demonstrated a positive correlation between APE1 and CD8⁺ T cell proportion in the SCLC patients (r=0.27, P<0.05). In vitro experiments showed that the overexpression (OE) cells (OEAPE1 and OELIGⅠ) exhibited reduced sensitivity to cisplatin than the NC group (P<0.05). Animal experiments indicated that cisplatin+E3330 significant inhibited xenograft tumor growth, indicating enhanced therapeutic efficacy（P<0.01）.

High expression of APE1, POLβ, and LIGⅠ in the BER pathway indicates poor prognosis and low DFS rate in SCLC patients. High expression of APE1 is positively correlated with CD8⁺T cells, and can be used as an auxiliary marker for SCLC immunotherapy.

To identify the key biomarkers for diagnosing chronic obstructive pulmonary disease (COPD) complicated with pulmonary arterial hypertension (PAH) using bioinformatics, and validate their clinical significance.

High-throughput sequencing data analysis was employed to identify differentially expressed genes (DEGs) in COPD-PAH. Functional enrichment analysis was then conducted to explore the biological functions of these DEGs. Machine learning methods, including least absolute shrinkage and selection operator (LASSO), random forest (RF), and support vector machine-recursive feature elimination (SVM-RFE), were utilized to screen 5 potential biomarkers. Single-cell analysis was performed to reveal the expression patterns of these key genes in macrophages. The clinical significance of these biomarkers was further validated using peripheral blood mononuclear cells (PBMC) data. A mouse model of COPD-PAH was established using hypoxia exposure. Sixteen mice (either sexes, 8 weeks old, weighing 20~22 g) were randomly divided into a hypoxia group [O₂ (10.0±0.5)%, COPD-PAH, n=8] and a normoxia group (COPD, n=8). Immunofluorescence assay was used to label the key biomarkers, and their expression levels were quantified.

A total of 28 DEGs (|Log₂FC|≥2, P<0.05) were identified in COPD-PAH patients. Functional enrichment analysis indicated that DEGs in COPD were primarily associated with major histocompatibility complex (MHC) Ⅱ and cell division, and involved in lysosomes, oxidative phosphorylation, and cell cycle pathways (P<0.05). Machine learning identified 5 potential biomarkers (GRN, KLF4, SHTN1, LRP1, and GPNMB), and subsequent single-cell analysis revealed that these markers exhibited reverse expression patterns during disease progression. A nomogram model constructed based on PBMC data yielded an area under the curve (AUC) of 0.907 in diagnosing COPD-PAH. GRN, KLF4, SHTN1, LRP1 and GPNMB were significantly upregulated in the COPD-PAH group (P<0.05).

GRN, KLF4, SHTN1, LRP1 and GPNMB are identified as key biomarkers for the prediction and diagnosis of COPD-PAH, which providing new insights for the clinical and treatment of the condition.

To investigate the factors influencing the efficacy of intravesical Bacille Calmette-Guérin (BCG) instillation after transurethral resection of bladder tumor (TURBT) in patients with intermediate- and high-risk non-muscle invasive bladder cancer (NMIBC), and to construct a prediction model for recurrence after BCG treatment.

A retrospective cohort study was conducted on the subjected patients diagnosed with intermediate- and high-risk NMIBC undergoing TURBT followed by standard BCG instillation. The 110 patients treated in Department of Urology of Army Medical Center of PLA from January 2018 to December 2023 were assigned into a training set, while the 52 patients treated at Department of Urology of General Hospital of Central Theater Command from January 2015 to December 2020 were into an external validation set. A total of 17 variables were included and analyzed. Univariate and multivariate Cox regression analyses were performed to identify factors associated with recurrence after BCG instillation, and nomograms were plotted to predict 1-year, 3-year, and 5-year recurrence-free survival (RFS). Calibration curve, decision curve analysis (DCA), and receiver operating characteristic (ROC) curve analysis were conducted for internal and external validation to evaluate the predictive performance and clinical utility of the model.

In the training set, 26 patients (23.64%) experienced recurrence during the follow-up period, with a median RFS of 32.00 (18.00~50.50) months. Univariate Cox regression analysis suggested that platelet count, eosinophil to lymphocyte ratio (ELR), neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), systemic immune inflammation (SII) index, and neutrophil-monocyte to lymphocyte ratio (NMLR), pathological T1 stage (pT1) tumor and hemoglobin, albumin, lymphocyte, and platelet (HALP) score were potential factors influencing recurrence after BCG instillation. Multivariate Cox regression analysis identified high HALP score (HR=0.185, 95%CI: 0.046~0.736, P=0.017) as an independent protective factor, while high ELR (HR=3.599, 95%CI: 1.505~8.608, P=0.004) and pT1 stage (HR=3.240, 95%CI: 1.191~8.818, P=0.021) were independent risk factors for recurrence. Based on this, a nomogram prediction model was constructed. The calibration curves demonstrated good agreement between predicted and actual 1-, 3-, and 5-year recurrence risks. Decision curve analysis indicated clinical utility across a wide threshold probability range. In the training set, the model showed strong predictive performance for 1- (AUC=0.842), 3- (AUC=0.847), and 5-year (AUC=0.887) recurrence risks, which was further validated in the external cohort.

Higher HALP score prior to BCG instillation therapy is a protective factor against tumor recurrence, while higher ELR and pT1 stage are risk factors. Our nomogram prediction model based on HALP score, ELR and pathological T stage, can identify individuals at high risk of recurrence after BCG instillation therapy.

To investigate the risk factors for the occurrence of metabolic dysfunction-associated steatotic liver disease(MASLD) in lean individuals and construct a risk prediction model.

Based on the National Health and Nutrition Examination Survey(NHANES) database in the United States(from January 2017 to March 2020), 1 123 adult individuals were included in this study. Then the participants were randomly divided into a training set(n=561) and a validation set(n=562) through simple random sampling. Data on their demographics, anthropometrics, lifestyle, underlying diseases, and laboratory test results were collected. LASSO regression analysis was used to screen potential variables in the training set, and multivariate logistic regression was employed to identify independent risk factors for lean MASLD. Based on these risk factors, a prediction model for lean MASLD was constructed(LMPM). To evaluate the clinical value of the LMPM, it was compared with two commonly used prediction models for non-alcoholic fatty liver disease, the fatty liver index(FLI) and the hepatic steatosis index(HSI). The performance of the model was evaluated and internally validated using the area under the receiver operating characteristic curve(AUC), net reclassification index(NRI), integrated discrimination improvement(IDI), calibration curve, decision curve, and clinical impact curve.

Age, waist circumference, and triglycerides(TG) were identified as independent risk factors for the development of MASLD in lean individuals. The LMPM, constructed based on these indicators, demonstrated good discriminative ability in both the training and validation sets, with AUC values of 0.86(95%CI: 0.82~0.89) and 0.81(95%CI: 0.77~0.85), respectively, which were significantly better than those of FLI [training set: AUC=0.83(95%CI: 0.79~0.87); validation set: AUC=0.74(95%CI: 0.70~0.79)] and HSI [training set: AUC=0.71(95%CI: 0.66~0.76); validation set: AUC=0.71(95%CI: 0.65~0.76)]. Compared with FLI and HSI, the LMPM showed improvements in NRI and IDI in both the training and validation sets. The calibration curve demonstrated its high accuracy, and both the decision curve analysis and the clinical impact curve analysis indicated that the LMPM provided greater clinical benefits.

Age, waist circumference, and TG are independent risk factors for lean MASLD. Based on these factors, a prediction model, named LMPM, is developed to assess the risk of MASLD in lean individuals, which exhibits good predictive performance and has certain guiding significance for the timely identification of high-risk populations.

To analyze the diversity and composition of gut microbiota in individuals with circadian rhythm disruption and their correlation with cognition.

Night shift workers and regular shift workers were subjected from our hospital during August 2022 and October 2024. The participants with circadian rhythm disorders were assigned into an experimental group (n=24), and those with normal circadian rhythms were into a control group (n=24). Their height, weight, age, gender, body mass index (BMI) and fresh fecal samples were collected, and Montreal Cognitive Assessment (MoCA) and Mini-Mental State Examination (MMSE) were used to evaluate their mental status. Metagenomics, Alpha and Beta diversity analyses, Linear Discriminant Analysis Effect Size (LEfSe), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were employed to investigate the diversity and function characteristics of gut microbiota in the participants.

There were no statistical differences between the 2 groups in baseline data, such as height, weight, gender, age, and BMI (P>0.05). Alpha diversity analysis indicated that no statistical differences were observed in the ACE, Chao1, Shannon, or Simpson indices between the 2 groups, while beta diversity analysis revealed significant differences (P<0.01), suggesting different structure of gut microbiota between them. In the experimental group, the abundance of Faecalibacterium prausnitzii and Agathobacter rectalis was decreased, while that of Escherichia coli and Phocaeicola vulgaratus was increased, with significant differences when compared with the control group (P<0.05). Additionally, KEGG functional analysis showed that the experimental group had obviously higher expression levels in Th17 cell differentiation and the IL-17 signaling pathway than the control group (P<0.05). Agathobacter rectalis and Faecalibacterium prausnitzii were positively correlated with MoCA score and MMSE score (P<0.05, P<0.01). Agathobacter rectalis was negatively correlated with the IL-17 signaling pathway and Th17 cell differentiation.

Individuals with circadian rhythm disorders have significant changes in the structure and function of gut microbiota when compared to those with normal circadian rhythms. Agathobacter rectalis may be involved in the regulation of the IL-17 signaling pathway and differentiation of Th17 cells, thereby possibly impacting the increases of cognitive score related to circadian rhythm disorders.

To compare the safety and efficacy of curettage and their combination with uterine artery embolization (UAE) in the treatment of cesarean scar pregnancy (CSP) patients with a low score (≤4) in the ultrasound quantification scoring system.

Based on our inclusion and exclusion criteria of this randomized controlled study, the women with CSP who had an ultrasonic quantitative score ≤4 and were treated in our department from May 2020 to August 2023 were enrolled, and then randomly divided into a curettage group (n=48) and a UAE combination group (n=47) in a ratio of 1∶1. General information, intraoperative conditions, and use of rescue measures within 3 months after operation were collected in the 2 groups of patients. All the patients were followed up until October 2024 to observe the pregnancy outcomes and determine the impact on the menstrual volume after the resumption of normal menstruation.

The patients from the both groups completed the follow-up. Except for the maximum gestational sac diameter, there were no significant differences in other baseline data between the 2 groups, and the curettage group had notably more patients having a gestational sac diameter ≤25 mm than the combination group [37 (77.1%) vs 27 (57.4%), P<0.05]. No statistical differences were observed between the 2 groups in the intraoperative bleeding volume and use of rescue measures within 3 months after surgery. The combination group had obviously more patients with reduced menstrual volume after the resumption of normal menstruation than the dilation and curettage group [30 (63.8%) vs 13 (27.1%), P<0.001]. There were no statistically differences in pregnancy outcomes and the number of days to resume menstruation between the 2 groups.

For CSP patients with a score of ≤4 in the ultrasound quantification scoring system, curettage show no significant difference in therapeutic effectiveness, and even have better efficacy and safety when compared with curettage combined with UAE.

To survey conflict occurrence in surrogate decision makers for respiratory intensive care unit (RICU) patients with mechanical ventilation and explore its influencing factors.

A total of 101 surrogate decision-makers of mechanical ventilation patients in RICU were subjected from the Second Affiliated Hospital of Army Medical University from June 2024 to December 2024 by convenience sampling. General data questionnaire, Preparation for Decision Making Scale and Decisional Conflict Scale were used to conduct questionnaire survey. One-way ANOVA, Pearson correlation analysis and multiple linear regression analysis were employed to identify the influencing factors.

The total score of decision readiness was 64.14±22.35, the total score of decision conflict was 35.18±26.47, and the incidence of decision conflict was 69.31% in the participants. Univariate analysis showed that there were statistically differences in whether knowing about prognosis, relationship with patients, awareness of patients’ ideas about treatment, and understanding of information related to mechanical ventilation among the participants (P<0.05). Pearson correlation analysis indicated that decision readiness was negatively correlated with decision conflict and its 3 dimensions (P<0.001). The results of multiple linear regression analysis revealed that understanding of information related to mechanical ventilation and decision readiness were influencing factors for the decision conflict of the participants (P<0.001).

The decision conflict level is quite higher in the surrogate decision makers for RICU patients with mechanical ventilation, and the level is negatively correlated with the understanding of mechanical ventilation related information and decision readiness.

To investigate the effects of joint function screening and correction on intervention efficacy of prevention and assessment score of training injuries in new recruits.

A randomized controlled study was conducted on 265 new recruits subjected from two organizational units of an army unit with cluster sampling. Based on entire organizational unit, the participants were randomly divided into a control group (n=132) and an experimental group (n=133). The experimental group received joint function screening and corrective movement training, which was subsequently applied in the new recruit training, while the control group underwent training according to conventional methods. Joint function were collected before and after training. The demographic data, assessment score of training, and incidence of training injuries were collected through the participant’s own organizational unit. Receiver operating characteristic (ROC) curve was plotted to evaluate the efficacy of joint function screening in predicting training injuries, and binary logistic regression and general linear regression analyses were applied to verify the correlation of joint function screening score with training injuries and assessment score of training.

After new training, the score of joint function screening was significantly higher in the experimental group than the control group (16.62±1.87 vs 14.92±2.58, P<0.001). And the score was obviously increased in the experimental group (16.62±1.87 vs 12.82±1.98, P<0.001) and the control group (14.92±2.58 vs 12.95±1.81, P<0.001) when compared with the corresponding score before training. The area under the ROC curve (AUC) of joint function screening in predicting training injuries was 0.762 (95%CI: 0.694~0.830), indicating good predictive efficacy. During the new training process, the incidence of training injuries in the experimental group (13.53%) was significantly lower than that in the control group (24.24%, Chi-square=4.963, P=0.026). Binary logistic regression analysis showed that the pre-training assessment score of joint function screening was an important influencing factor for training injuries in new recruits (OR=0.552, 95%CI: 0.413~0.660, P<0.001). The experimental group obtained notably higher mean assessment score than the control group [733.00（716.00，752.75） vs 728.79（710.46，744.28）, P=0.027]. Linear regression analysis revealed a correlation between post-training score of joint function screening and the assessment score of newly trained personnel (P<0.001).

Joint function screening and correction for newly trained personnel can effectively prevent training-related injuries during the new training period, and correcting joint function through training can effectively improve the assessment score of newly trained personnel.