To comparatively evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) versus conventional microbial culture in spinal infections.

A cross-section design was conducted on 82 consecutive patients with suspected spinal infections treated between February 2022 and January 2024 at Jiangbei Branch of First Affiliated Hospital of Army Medical University （Third Military Medical University）. Microbiological culture, histopathological examination, and mNGS results from infected specimens were analyzed. Clinical diagnosis, primarily based on clinical manifestations, laboratory tests and radiologic features combined with medical history, was defined as the gold standard, and then the diagnostic performance, including sensitivity and specificity, were compared between mNGS and microbial culture.

Among the 82 patients, definitive microbiological evidence was identified in 70 cases, and mNGS demonstrated a significantly higher detection rate than microbial culture (64 vs 36 cases, 78.05% vs 43.9%, P<0.05). mNGS also obtained obviously higher sensitivity, accuracy, and negative predictive value (NPV), and notably lower positive predictive value (PPV) when compared to conventional microbial culture (all P<0.05). When stratified by infection type, mNGS obtained significantly higher sensitivity and accuracy compared to microbial culture in tuberculous spinal infections (P<0.05). For non-tuberculous spinal infections, mNGS also showed superior sensitivity to microbial culture (P<0.05).

In patients with spinal infections, mNGS demonstrates a significantly higher pathogen detection rate than conventional microbial culture. This technique can provide early and broad-spectrum pathogenic microbiological evidence for spinal infection.

To develop and validate a prediction model for risk of amputation in patients with diabetic foot ulcers (DFU) based on systematic review and meta-analysis.

The studies on the risk factors of amputation in DFU patients was retrieved by using subject words+free words. After screening, 37 cohort studies were finally included, and the Newcastle-Ottawa scale (NOS) was used for quality evaluation. Meta-analysis was performed on the risk factors of amputation in DFU. Then a prediction model for DFU amputation risk were constructed based on the statistically significant risk factors in the meta-analysis. The corresponding β value was calculated based on the combined odds ratio (OR) value of each risk factor, and each risk factor was scored to establish a scoring system model. The clinical data of 453 DFU patients hospitalized in our department from 2021 to 2023 were collected as a validation cohort. Receiver operating characteristic (ROC) curve analysis was used to evaluate the model performance. The area under the curve (AUC) was calculated, and the optimal cutoff score was determined by calculation of the maximum Youden index through sensitivity and specificity.

Our meta-analysis showed a cumulative amputation rate of approximately 34.65% in 11 779 DFU patients. The final risk prediction models include gangrene [OR=11.92 (5.86~24.24)], ulcer depth [OR=4.93 (2.52~9.64)], osteomyelitis [OR=3.19 (2.36~4.29)], previous amputation history [OR=3.19 (2.00~5.09)] and lower extremity arterial disease [OR=3.10 (2.31~4.17)]. According to the weights of each risk factor, the total score of the model is 76, and the optimal cut-off score is 36.5. The prediction model performed well, with an AUC value of 0.864 (0.824, 0.903), a sensitivity of 0.743, a specificity of 0.859, and an accuracy rate of 83.00%.

A prediction model for DFU amputation risk is developed based on risk factor scoring, and has good discrimination and calibration, providing effective scientific basis for clinical research and clinical decision-making related to DFU amputation.

To analyze the disease burden, changing trends, and differences of atrial fibrillation (AF)/atrial flutter (AFL) globally and in China and Japan from 1990 to 2021, and to predict their future trends, aiming to provide references for health decision-making.

Based on the Global Burden of Disease Study Database 2021 (GBD 2021), we extracted age-standardized prevalence rate (ASPR) and age-standardized disability-adjusted life year rate (ASDALYR) data for AF/AFL by sex globally, in China, and Japan. The estimated annual percentage change (EAPC) was used to assess the trends. Joinpoint regression analysis and the Bayesian age-period-cohort (BAPC) model were employed for trend analysis and prediction.

From 1990 to 2021, the ASPR and ASDALYR for AF/AFL in males were increased significantly globally, with EAPC of 0.05 (95%CI: 0.01~0.08) and 0.09 (95%CI: 0.08~0.11), respectively. Changes were significantly declined in females,with EAPC of -0.11(95%CI: -0.14~-0.07)and -0.10(95%CI:-0.12~-0.07). In China, the ASPR for AF/AFL were increased in both males and females, with those of males more notably (EAPC=0.77, 95%CI: 0.65~0.88). However, the ASDALYR for AF/AFL showed gender divergence, with an increase in males (EAPC=0.40, 95%CI: 0.30~0.49) while a decrease in females (EAPC=-0.55, 95%CI: -0.67~-0.44). In Japan, both the ASPR and ASDALYR for males and females showed continuous declines, and the reduction was more pronounced among females (ASPR EAPC=-1.77, 95%CI:-2.32~-1.22; ASDALYR EAPC=-1.73, 95%CI:-2.11~-1.35). Joinpoint regression analysis showed that from 1990 to 2021, for the ASDALYR of AF/AFL, the average annual percentage change (AAPC) was 0.28% (P<0.001) for Chinese males and -0.37% (P<0.001) for Chinese females, while the AAPC was -0.70% (P<0.001) and -1.43% (P<0.001) for Japanese males and females. BAPC model revealed that by 2036, the ASDALYR for Chinese males is predicted to increase from 91.45 per 100 000 in 2022 to 101.11 per 100 000, and for females is from 88.85 per 100 000 to 100.98 per 100 000. For Japanese males, the ASDALYR is projected to increase slightly from 88.79 to 89.86 per 100 000, while for females, it is projected decrease slightly from 41.13 to 39.67 per 100 000, indicating only minor fluctuations in the ASDALYR for both Japanese males and females.

The disease burden of AF/AFL continues to increase globally and in China. So, Japan’s lifestyle and health policies are worth considering, and more scientific and effective public health policies and clinical intervention strategies should be formulated and implemented. Countermeasure　The relevant government agencies should promote the transformation of the food industry through the policy-market mechanism, carry out activities related to national health management, and continuously optimize the AF/AFL management model of medical and health institutions to effectively cope with this disease burden change.

To develop a preoperative question prompt list (QPL) for older patients with benign prostatic hyperplasia (BPH) and evaluate its effectiveness in application.

This trial adopted a randomized controlled design. The QPL was developed by literature review, expert discussions, and Delphi consultation. Convenience sampling was used to subject 76 older BPH inpatients treated in our department, and then they were randomly divided into control (routine communication, n=38) and intervention (QPL-assisted communication, n=38) groups. Number of the questions patient asking, communication duration, information recall, and communication quality were compared between the 2 groups.

In the 2 rounds of expert consultation, the response rate of questionnaire was 94.44% and 100%, the authority coefficient was 0.89 and 0.93, the coefficient of variation was 0.05~0.22 and 0~0.11, and Kendall’s coefficients was 0.645 (Chi-square=87.782, P<0.001) and 0.733 (Chi-square=74.789, P<0.001), respectively. The final QPL included 3 themes and 7 questions. The intervention group asked more questions (4.03±1.89 vs 2.11±1.27, P<0.05) but spent similar time for communication (8.18±2.11 vs 7.67±1.72 min, P>0.05). At 1 d before discharge, better information recall (8.74±1.12 vs 6.49±1.68, P<0.001) and communication quality (60.06±6.25 vs 54.86±7.98, P<0.05) were observed in the intervention group when compared with the control group.

Our developed preoperative communication QPL is of scientificalness and effectiveness for elderly BPH patients. This tool can not only encourage question-asking behavior, but also improve information recall and communication quality in the patients.

To reveal the molecular mechanism by which nuclear epidermal growth factor receptor (nEGFR) synergistically regulates the expression of cell migration-inducing protein (CEMIP) by forming a complex with the transcription factor Yin Yang 1 (YY1), and to investigate the biological functions of the nEGFR-YY1-CEMIP signaling axis in invasion of hepatocellular carcinoma (HCC).

After HCC cells were serum-starved for 24 h, the cells were treated with 100 ng/mL EGF. Thus, the cells were divided into a control group and EGF-treated groups at different time points. Nuclear expression and localization changes of EGFR were detected by Western blotting and immunofluorescence (IF). To investigate the interaction between nEGFR and YY1, their nuclear colocalization and interaction were examined by IF and co-immunoprecipitation (Co-IP), respectively. Transcriptional profiling was performed using RNA sequencing (RNA-seq) to identify differentially expressed genes at the genome-wide level. Combined with Gene Ontology (GO) functional enrichment analysis and transcription factor binding profiles via using the JASPAR database, CEMIP was identified as a candidate target gene. To validate the regulatory mechanism, the following experimental groups were established, Control, EGF, siYY1, and siYY1+EGF. The expression of CEMIP at protein and mRNA levels was detected by Western blotting and RT-qPCR. To elucidate the molecular mechanism of nEGFR/YY1 binding to the CEMIP promoter, the control and EGF-treated groups were established. Chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) was performed to assess the enrichment of nEGFR/YY1 at the CEMIP promoter region. Luciferase reporter assay was conducted following transfection with either wild-type EGFR (EGFR-WT), nuclear localization-deficient mutant (EGFR-dNLS), YY1 overexpression plasmid (YY1-OE), or dominant-negative YY1 mutant (YY1-DN) to evaluate changes in promoter activity. Subsequently, cell migration and invasion capabilities were evaluated using scratch wound healing assay and Transwell assay, while hyaluronic acid (HA) level was quantified by ELISA. The expression of matrix metalloproteinases (MMP2/9) was analyzed via Western blotting to assess the regulatory role of the nEGFR/YY1-CEMIP axis in the migration and invasion of HCC cells. By analyzing the CEMIP expression profiles in HCC patients from National Center for Biotechnology Information (NCBI) public databases, its potential association with tumor metastasis risk was validated.

Western blotting and IF demonstrated that EGF treatment significantly induced nuclear translocation of EGFR, peaking at 30 min (P<0.001). Co-IP and IF assays indicated both physical interaction and nuclear co-localization between nEGFR and YY1. RNA-seq analysis identified CEMIP as a significantly differentially expressed gene. GO enrichment analysis revealed that CEMIP was significantly enriched in biological processes related to cell invasion promotion. JASPAR prediction identified conserved YY1 potential binding region within the CEMIP promoter region. Western blot and RT-qPCR analyses confirmed that EGF treatment up-regulated CEMIP at both protein and mRNA levels (P<0.05). Notably, YY1 knockdown significantly suppressed CEMIP expression, while exogenous EGF supplementation restored CEMIP level in YY1-deficient cells (P<0.05). ChIP-qPCR analysis demonstrated specific enrichment of the nEGFR/YY1 complex at the CEMIP promoter region, with EGF stimulation significantly enhancing its binding affinity (P<0.001). Luciferase reporter assay confirmed that nEGFR/YY1 robustly enhanced CEMIP promoter activity (P<0.01), while either the EGFR-dNLS or the YY1-DN substantially attenuated this transcriptional activation. Functional phenotyping showed that the nEGFR/YY1-CEMIP axis significantly enhanced the migration and invasion of HCC cells by promoting HA catabolism and up-regulating MMP2/9 expression (P<0.05). Analysis of NCBI datasets revealed that CEMIP expression was significantly up-regulated in HCC tumor tissues than adjacent normal tissues (P<0.001). Moreover, HCC patients with elevated CEMIP expression exhibited higher risk of metastasis (P<0.001).

nEGFR promotes HCC invasion by forming a transcriptional complex with YY1 to cooperatively activate CEMIP expression.

Toreveal the mechanism by which the programmed death-ligand 1 （PD-L1）- protein tyrosine phosphatase 1B （PTP1B）-focal adhesion kinase （FAK） signaling axis promotes the progression of hepatocellular carcinoma （HCC） and elucidate its effector functions in HCC.

GEPIA database was used to plot a 10-year survival curve for PD-L1 and FAK expression levels in HCC patients. Immunohistochemical （IHC） staining was utilized to analyze the relative expression levels of PD-L1 and FAK phosphorylated at the Y397 site ［p-FAK（Y397）］ in HCC tissues，and the results were compared to those in the adjacent non-tumor tissues. Subsequently，endogenous PD-L1 expression was detected with Western blotting in HCC cell lines with low （SNU-387） and high （Hep3B） PD-L1 expression levels. After lentivirus-transduced SNU-387^PDL1+ and Hep3B^PDL1-cells were constructed，the effect of high and low expression of PD-L1 on the expression of p-FAK（Y397） with Western blotting. To elucidate the functional mechanism of FAK in HCC，functional rescue experiments were performed by administering a FAK inhibitor to SNU-387^PDL1+ cells and a FAK activator to Hep3B^PDL1- cells，combined with wound healing scratch assay，Transwell invasion assay，EdU proliferation assay，and colony formation assay to evaluate tumor malignant effects. The GENEMANIA database predicted functional interactions between protein tyrosine phosphatase 1B （PTP1B），PD-L1，and FAK. IHC staining was performed to analyze the correlation among PD-L1，PTP1B，and p-FAK（Y397） expression. Co-immunoprecipitation （Co-IP） and indirect immunofluorescence （IF） were applied to validate the interaction between PD-L1 and PTP1B. Western blotting was utilized to confirm the regulatory relationship between PD-L1 and PTP1B. In vitro PTP1B phosphatase activity assay measured the changes in PTP1B activity. Subsequently，Western blotting was used to screen cell lines with high endogenous PTP1B expression （SNU-387） and low endogenous PTP1B expression （Hep3B）. Furthermore，Hep3B^PTP1B+ and SNU-387^PTP1B- cell lines were generated，and then p-FAK（Y397） levels were then detected in these modified cell lines，and the aforementioned functional effect assays （migration，invasion，proliferation and colony formation） and rescue experiments were repeated. Furthermore，Western blotting was employed to detect changes in downstream signaling pathways following enhancement or attenuation of p-FAK（Y397） in SNU-387 and Hep3B cells.

IHC staining revealed a positive correlation between PD-L1 and p-FAK（Y397） expression in HCC tissues （95%CI：1.065~3.801，P<0.01）. In SNU-387^PDL1+ cells，PD-L1 overexpression significantly enhanced phosphorylation at the FAK Y397 site （P<0.01） and increased cell migration，invasion，proliferation，and colony formation capabilities （P<0.01），and these effects could be reversed by FAK inhibitor treatment （P<0.05）. Conversely，in Hep3B^PDL1- cells，PD-L1 knockdown significantly reduced FAK Y397 phosphorylation （P<0.01） and decreased cell migration，invasion，proliferation，and colony formation abilities （P<0.01），and these effects were restored by FAK activator treatment （P<0.05）. IHC staining further showed a negative correlation between PTP1B expression and both PD-L1 and p-FAK（Y397） in HCC tissues （95%CI：1.886~3.514，P<0.05）. Co-IP and IF assays confirmed a direct interaction between PD-L1 and PTP1B，with PD-L1 suppressing PTP1B expression level and reducing its activity （P<0.01）. In SNU-387^PTP1B- cells，PTP1B knockdown significantly increased FAK Y397 phosphorylation （P<0.01） and enhanced cell migration，invasion，proliferation，and colony formation （P<0.01），and these effects were reversed by FAK inhibitor （P<0.05）. While in Hep3B^PTP1B+ cells，PTP1B overexpression significantly decreased FAK Y397 phosphorylation （P<0.01） and reduced cell migration，invasion，proliferation，and colony formation （P<0.01），and those effects were restored by FAK activator treatment （P<0.05）. Furthermore，enhanced phosphorylation at the FAK Y397 site in SNU-387 cells activated downstream PI3K/AKT and MEK/ERK signaling pathways （P<0.01），whereas inhibition of FAK（Y397） phosphorylation in Hep3B cells attenuated the activation of these signaling pathways （P<0.01）.

PD-L1 activates FAK by suppressing PTP1B，thereby promoting migration，invasion，and proliferation in HCC.

To investigate the effects of glutathione peroxidase 7 (GPX7) on the proliferation and senescence of hepatocellular carcinoma (HCC) cells and its potential value as a therapeutic target.

Clinical and transcriptomic data from HCC patients in The Cancer Genome Atlas (TCGA) database were analyzed to evaluate the relationship between GPX7 expression and prognosis. Western blotting, qRT-PCR, CCK-8 assay, EdU staining, and SA-β-Gal staining were employed to compare the expression and function of GPX7 in HCC cell lines and normal liver cells. A stable GPX7-knockdown Hep3B cell line was established, and transcriptomic changes were analyzed using RNA-seq. The PI3K inhibitor LY294002 and its activator 740 Y-P were utilized to determine the role of GPX7 in regulating cell senescence via the PI3K-AKT pathway. An orthotopic xenograft model was constructed using 18 male nude mice (6 weeks old, 18~22 g) to observe tumor growth. Tumor and adjacent tissue samples from 56 HCC patients (obtained from the Eighth Medical Center of Chinese PLA General Hospital between January 2012 and December 2022) were analyzed to assess the relationship between GPX7 expression and clinicopathological features.

TCGA data analysis revealed that GPX7 was significantly highly expressed in HCC tumor tissues (P<0.01), and the patients with high GPX7 expression had obviously shorter overall survival (OS) (P=0.018). Western blotting and qRT-PCR showed that the expression level of GPX7 was notably higher in the HCC cell lines than the normal liver cells (P<0.01). After GPX7 knockdown, the CCK-8 absorbance values of HCC cells were significantly reduced (P<0.01), the proportion of EdU-positive cells was obviously decreased (P<0.01), and the proportion of SA-β-Gal positive cells was notably increased (P<0.01). RNA-seq analysis revealed that differentially expressed genes were significantly enriched in the PI3K-AKT signaling pathway (P<0.05). The PI3K inhibitor LY294002 significantly enhanced the GPX7 knockdown-induced cell senescence (P<0.01), while the activator 740 Y-P partially reversed this effect (P<0.05). In the xenograft model, GPX7 knockdown significantly inhibited tumor growth in nude mice (P<0.01), and the PI3K activator 740 Y-P partially reversed this effect (P<0.01). Clinical sample analysis displayed that high GPX7 expression was negatively correlated with tumor differentiation (P<0.05), and its high expression was associated with shortened OS (HR=2.61, 95%CI: 1.26~5.39, P=0.005).

GPX7 regulates cell proliferation and senescence through the PI3K-AKT pathway, and may serve as a potential therapeutic target for HCC treatment.

To investigate the mechanism by which INKA2-AS1 promotes hepatocellular carcinoma(HCC) migration, invasion, and macrophage-related immunosuppression.

Based on the Cancer Genome Atlas Program (TCGA)-HCC data and Genotype-Tissue Expression (GTEx) databases, differentially expressed genes were analyzed in 374 HCC tissues and 160 normal liver tissues. Survival analysis was used to investigate the relationship between INKA2-AS1 and the prognosis of HCC patients. Cox regression analysis was performed on INKA2-AS1. The relationship between INKA2-AS1 and immune infiltration in HCC tissues was analyzed using the Tumor Immune Estimation Resource Database. A total of 90 HCC patients were recruited from the Department of Hepatobiliary Surgery, Chongqing Hygeia Hospital from January 2019 to January 2021. The HCC tissues and adjacent normal tissues were collected. Hepa 1-6 liver cancer cells were grouped into: normal control (NC), INKA2-AS1, siNC and siINKA2-AS1. INKA2-AS1 was overexpressed and silenced by transfection. The proliferation, migration and invasion of cells in each group were detected by CCK-8, EdU, TUNEL, wound healing and Transwell assays. Each group of Hepa 1-6 cells were co-cultured with macrophages to analyze their effects on macrophages. Twenty nude mice (SPF grade, 4 weeks old, 15~19 g, half male and half female) were randomly divided into 4 groups (n=5): NC, INKA2-AS1, siNC and siINKA2-AS1. Tumor-bearing nude mouse models were established by subcutaneous injection of INKA2-AS1 silenced and overexpressed Hepa1-6 cells. The effect of INKA2-AS1 on tumor volume and mass were detected. Macrophage immunity was analyzed by Western blot analysis of IL-10 and FAP protein levels in tumor tissues.

The level of INKA2-AS1 in HCC tissues was significantly higher than that in normal tissues(P<0.05). INKA2-AS1 was associated with the histological grade, pathological stage of HCC (P<0.05). The survival rate (P=0.05), disease-specific survival (P=0.036) and progression-free interval (P=0.024) of HCC patients with high INKA2-AS1 expression were significantly lower than those of patients with low INKA2-AS1 expression. The level of INKA2-AS1 in HCC tissues was significantly higher than that in adjacent normal tissues (P<0.05). Cox regression analysis showed that INKA2-AS1 was an independent predictor of OS in HCC patients (P=0.005). INKA2-AS1 expression was associated with immune cells such as T cells and macrophages. The proliferation, migration, and invasion abilities of the siINKA2-AS1 group were lower than those of the siNC group (P<0.05), the apoptotic rate was higher than that of the siNC group (P<0.05), and inhibited the expression of IL-10 and FAP at mRNA and protein levels in co-cultured macrophages (P<0.05). The proliferation, migration and invasion abilities of the INKA2-AS1 group were higher than those of the NC group (P<0.05), the apoptotic rate was lower than that of the NC group (P<0.05), and promoted the expression of IL-10 and FAP at mRNA and protein levels in co-cultured macrophages (P<0.05). The tumor volume and mass of the siINKA2-AS1 group were lower than those of the siNC group (P<0.05), while the tumor volume and mass of the INKA2-AS1 group were higher than those of the NC group (P<0.01). The levels of IL-10 and FAP proteins in tumor tissues of the siINKA2-AS1 group were lower than those of the siNC group (P<0.05), while the levels of IL-10 and FAP proteins were increased (P<0.05).

INKA2-AS1 increment is related to poor prognosis of HCC patients, and the molecule promotes the proliferation and metastasis of HCC cells and induces the transformation of tumor-associated macrophages.

To investigate the value of enhanced MRI image features combined with apparent diffusion coefficient(ADC) in the differential diagnosis of primary hepatic mucosa-associated lymphoid tissue(MALT) lymphoma and hepatocellular carcinoma(HCC).

The clinical data of 10 patients with pathology-confirmed primary MALT lymphoma admitted in our hospital from January 1997 to May 2025 were retrospectively collected. Their enhanced MRI images were analyzed, mainly including the number of lesions, morphology, signal characteristics, enhancement methods, and presence or absence of vessel floating signs, and the ADC value was calculated in order to screen statistically significant variables. Firth regression analysis was used to screen independent predictors of primary MALT lymphoma in the liver. Receiver operating characteristic(ROC) curve was plotted, and the area under the curve(AUC) was calculated to evaluate the diagnostic efficacy.

Compared with the HCC group, the primary MALT lymphoma group had significantly higher incidence in the females(P=0.026), alpha-fetoprotein(AFP) <8 ng/mL(P<0.001), lower T2WI signal(P=0.001), lower ADC value(P<0.001), and mostly continuously enhanced lesions(P<0.001), and more vessel floating signs(P=0.008). Univariate Firth regression analysis showed that sex(95%CI: 1.36~52.10, P=0.020), T2 value(95%CI: 3.89~5 121.00, P<0.001), AFP(95%CI: 0~0.21, P<0.001), ADC value(95%CI: 0~0.21, P<0.001), and dynamic reinforcement(95%CI: 0~0.23, P<0.001), and vessel floating sign(95%CI: 0~0.40, P=0.004) were diagnostic predictors for primary MALT lymphoma of the liver. Multivariate Firth regression analysis indicated that ADC value(95%CI: 0~0.62, P=0.006), AFP(95%CI: 0~0.047, P=0.01), and dynamic reinforcement(95%CI: 0~0.39, P=0.005) were independent predictors for primary MALT in the liver. ROC curve analysis indicated that the AUC value of ADC value in diagnosing primary MALT lymphoma was 0.96, with a sensitivity of 0.8 and a specificity of 1.0, and the AUC value of AFP level in diagnosing primary MALT lymphoma was 0.85, with a sensitivity of 1.0 and a specificity of 0.7. The AUC value of dynamic reinforcement in diagnosing primary MALT lymphoma was 0.85, with a sensitivity of 0.8, and a specificity of 0.9.

Dynamic reinforcement and ADC value can be used as differential markers for primary MALT lymphoma and HCC, providing reliable reference for clinical preoperative evaluation.