Shuilong Guo

Background: Ribonucleic acid export 1 (RAE1) autoantibody may have good potential for early detection of gastric cancer (GC). However, the carcinogenicity of RAE1 in GC remains unknown. We aimed to explore the role and the potential mechanism for RAE1 in the carcinogenesis of GC. Methods: Immunohistochemical assay was applied to analyze the expression of RAE1 in GC and precancerous lesion (PL) tissues and its relationship with clinical characteristics. The effects of RAE1 on proliferation, migration, apoptosis, and cell cycle were explored by constructing RAE1 knockdown and overexpression in GC cells. The effects of RAE1 knockdown on tumor growth were observed in a murine xenograft model. The signaling pathways involved in GC development that may be affected by RAE1 were investigated by transcriptome sequencing and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Results: Immunohistochemical expression of RAE1 was significantly higher in early GC patients than in PL and normal tissues, and the RAE1 expression was correlated with clinical stage (P In vitro, knockdown of RAE1 inhibited the proliferation and migration of GC cells with promoted apoptosis and arrested cell cycle in the S phase, while overexpression of RAE1 showed the opposite trend. In vivo, knockdown of RAE1 inhibited the growth of subcutaneous graft tumors in mice. Transcriptome sequencing and subsequent analysis of RAE1 knockdown cells revealed that the Hippo signaling pathway was activated by RAE1 knockdown. Conclusions: RAE1 promotes GC cells proliferation and migration and is associated with the inhibition of the Hippo signaling pathway, and may be a potential biomarker for early diagnosis and treatment of GC.

Heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) is associated with suboptimal glycopeptide treatment outcomes. However, conventional antimicrobial susceptibility testing fails to distinguish hVISA from vancomycin-susceptible Staphylococcus aureus (VSSA), potentially delaying effective therapy. The gold standard population analysis profiling with area under the curve (PAP-AUC) method is labor-intensive and time-consuming, necessitating rapid detection alternatives. This study introduces a microfluidic platform based on fluorescence-activated droplet sorting (FADS) for the rapid detection and isolation of hVISA. Through systematic optimization of key parameters such as droplet generation conditions, bacterial suspension concentration, fluorescence probe selection, fluorescence duration, and vancomycin screening concentration, we established a rapid hVISA screening system. The efficacy of this system was evaluated using the hVISA standard strain Mu3, followed by validation with 15 clinical hVISA isolates. Genomic analysis further elucidated the genetic basis of drug resistance in hVISA strains. We developed a high-throughput detection platform for hVISA by integrating a microfluidic system with FADS. Based on a Poisson distribution theoretical model, the bacterial suspension concentration was optimized to 106 CFU/mL, achieving a single-bacterium droplet encapsulation rate of approximately 30%. The selected fluorescent probe exhibited detectable fluorescence signals as early as 6 h post-cultivation, with stability maintained for at least 1 week. Drug concentration validation demonstrated that 4 μg/mL vancomycin effectively inhibited susceptible bacteria without interfering with fluorescence detection. This platform successfully isolated resistant subpopulations from the hVISA reference strain Mu3 within 12 h, and its efficacy was further validated using 15 clinical hVISA isolates. This study innovatively integrated a microfluidic detection platform and FADS to achieve rapid detection of hVISA. This technology reduced the detection time to just 12 h and significantly improved the enrichment efficiency of drug-resistant bacterial populations.

Background: Reprogramming glucose metabolism is a hallmark of human cancer during its occurrence and development. However, the comprehensive glycometabolism signature and underlying mechanism in CRC prognosis and immune response remind to be elucidated. Methods: A prognostic model derived from 297 glycometabolism-related genes (GRGs) was developed using LASSO-Cox and nomogram algorithms. Immune dysfunction between high-risk (Risk ^H ) and low-risk (Risk ^L ) groups was compared using CIBERSORT, TIMER, and TIDE analyses. The expression and function of key genes, including secreted frizzled-related protein 2 (SFRP2), were validated using PCR, western blotting, immunohistochemistry, transwell assays, and metastatic model in mice. Luciferase reporter and chromatin immunoprecipitation were used to determine the transcription regulation of ENO2 by TCF4. Results: More than half of the GRGs (152 out of 297) showed differential expression, mainly those associated with glycolysis and biosynthesis. The GRG-risk score outperformed other clinical indicators (AUC = 0.810) and served as an independent risk predictor ( P < 0.001, HR = 3.180). The Risk ^H group showed increased infiltration of immune cells and higher immune checkpoint expression. Mechanistically, SFRP2, a key gene in Risk ^H , promoted CRC glycolysis and metastasis via enolase 2 (ENO2) activation through the TCF4/β-catenin axis. Inhibiting ENO2 reversed SFRP2-induced metastasis. Coexpression of SFRP2 and ENO2 correlated with poorer survival and higher recurrence. Conclusion: The Risk ^H group is characterized by glycolysis overactivation and immune exclusion. SFRP2 and ENO2 have emerged as promising treatment targets for high-risk CRC patients.

Neonatal pneumonia is a leading cause of infant mortality worldwide; however, a lack of microbial profiling, especially of low-abundance species, makes accurate diagnosis challenging. Traditional methods can fail to capture the complexity of the neonatal respiratory microbiota, thereby obscuring its role in disease progression. Here, we describe a novel approach that combines high-throughput sequencing with droplet-based microfluidic cultivation to investigate microbiome shifts in neonates with pneumonia. Using 16S ribosomal RNA (rRNA) gene sequencing of 71 pneumonia cases and 49 controls, we identified 1009 genera, including 930 low-abundance taxa, which showed significant compositional differences between groups. Linear discriminant analysis effect size identified key pneumonia-associated genera, such as Streptococcus, Rothia, and Corynebacterium. Droplet-based cultivation recovered 299 strains from 94 taxa, including rare species and ESKAPE pathogens, thereby supporting targeted antimicrobial management. Host–pathogen interaction assays showed that Rothia and Corynebacterium induced inflammation in lung epithelial cells, likely via dysregulation of the PI3K-Akt pathway. Integrating these marker taxa with clinical factors, such as gestational age and delivery type, offers the potential for precise diagnosis and treatment. The recovery of diverse species can support the construction of a biobank of neonatal respiratory microbiota to advance mechanistic studies and therapeutic strategies.

Comamonas kerstersii is an environmental microorganism, and clinical infections caused by this species are rare. In this study, the strain CXZJT123 was isolated from gallbladder puncture fluid and identified using a comprehensive approach including morphological observation, mass spectrometry, biochemical identification, 16S rRNA sequencing, and whole-genome sequencing. The morphological characteristics of CXZJT123 were consistent with the Comamonas. Mass spectrometry initially identified the strain as Comamonas aquatica, while biochemical assays demonstrated key features aligning with the Comamonas genus. Phylogenetic analysis of 16S rRNA sequences placed CXZJT123 in the same clade as C. kerstersii (NR 025530.1). Whole-genome sequencing followed by gene annotation using the Non-Redundant Protein Database (NR) revealed the highest gene homology with C. kerstersii. Further genomic analysis using the Virulence Factors of Pathogenic Bacteria (VFDB) and the Comprehensive Antibiotic Research Database (CARD) identified the presence of virulence genes, including those encoding flagella (flg series) and biofilm formation (pil series), as well as antibiotic resistance genes such as macB, arlR, and evgS. This study presents the first reported isolation of C. kerstersii isolated from gallbladder puncture fluid. The findings contribute valuable insights into the identification, genomic characteristics, and potential pathogenicity of C. kerstersii, providing a useful reference for the diagnosis and management of related infections.

Staphylococcus aureus can develop antibiotic resistance and evade immune responses, causing infections in different body sites. However, the metabolic changes underlying this process are poorly understood. A variant strain, C1V, was derived from the parental strain C1 by exposing it to increasing concentrations of vancomycin in vitro. C1V exhibited a vancomycin-intermediate phenotype and physiological changes compared to C1. It showed higher survival rates than C1 when phagocytosed by Raw264.7 cells. Metabolomics analysis identified significant metabolic differences pre- and post-induction (C1 + SC1 vs. C1V + SC1V: 201 metabolites) as well as pre- and post-phagocytosis (C1 vs. SC1: 50 metabolites; C1V vs. SC1V: 95 metabolites). The variant strain had distinct morphological characteristics, decreased adhesion ability, impaired virulence, and enhanced resistance to phagocytosis compared to the parental strain. Differential metabolites may contribute to S. aureus ‘ resistance to antibiotics and phagocytosis, offering insights into potential strategies for altering vancomycin nonsusceptibility and enhancing phagocyte killing by manipulating bacterial metabolism.

Background: Rapamycin-insensitive companion of mammalian target of rapamycin (Rictor) protein is a core subunit of mammalian target of rapamycin complex 2, and is associated with cancer progression. However, the biological function of Rictor in cancer, particularly its clinical relevance in gastric cancer (GC) remains largely unknown.Methods: Rictor expression and its association with clinicopathologic characteristics in GC were analyzed by immunohistochemistry. Effect of Rictor and Caveolin-1 (Cav 1) on GC cells apoptosis was evaluated via overexpression experiment in vitro. Mechanisms of Rictor and Cav 1 in GC were explored through overexpression and knockdown, by immunofluorescence and western blot analyses.Results: Rictor was upregulated in GC, and mainly located in the cytoplasm of cancer cells. Moreover, higher Rictor levels were associated with worse prognosis. Rictor could inhibit GC cell apoptosis and promote cell growth in vitro. The results of immunofluorescence revealed that Cav 1 localized in GC cell membrane but did not co-localize with Rictor. Further, Rictor regulated apoptosis-related proteins, long non-coding RNAs and also activated cellular signaling, thereby positively regulating Cav 1 expression. This effect was attenuated by the Akt inhibitor ly294002. Cav 1 did not significantly affect the ability of Rictor to inhibit tumor cell apoptosis.Conclusions: Rictor is upregulated in GC and associated with worse prognosis. It inhibits tumor apoptosis and activates Cav 1 through the Akt signaling pathway to inhibit the apoptosis of GC cells. Rictor is, therefore, a promising prognostic biomarker and possible therapeutic target in GC patients.

Ovarian serous adenocarcinoma is one of the most common and fatal malignancies among women worldwide. The tumor microenvironment plays a critical role in tumor initiation, proliferation, and metastasis. Immune scores and stromal scores of the tumor microenvironment were determined using the ESTIMATE. Immune cell infiltration was assessed using TIMER and differentially expressed genes (DEGs) were determined using the R/Bioconductor package of edgeR. Survival analysis was carried out using a univariate Cox model and Kaplan-Meier survival, and gene functional information was obtained through Gene Ontology and KEGG pathway analysis. Survival analysis revealed 39 DEGs that significantly influenced the prognosis of ovarian serous adenocarcinoma patients and were correlated with immune cell abundance. Functional enrichment and protein-protein interaction network analyses further indicated that these genes are primarily involved in immune-related responses. Finally, we verified the prognostic value of these genes via GEO. The present results reveal the genes associated with the tumor microenvironment of ovarian adenocarcinoma, potentially providing prognostic information.

Gastric cancer (GC) is one of the most common and fatal malignancies worldwide. MicroRNAs (miRNAs) play a critical role in tumor initiation, proliferation, and metastasis of gastric cancer. miR193b has been identified as a tumor suppressor in a variety of tumor types; however, its role in gastric cancer is yet to be determined. Here, we found a significant downregulation of miR193b expression in both human gastric cancer tissues (p < 0.05) and human gastric cancer cell lines (p < 0.01). Furthermore, the expression level of miR193b correlated with the tumor type, tumor size, and clinical stage (p < 0.05). In vitro, miR193b overexpression inhibited cell survival and induced apoptosis in GC cell lines, indicating that miR193b plays a role in the development of gastric cancer. KRAS was verified as the target of miR193b, and KRAS overexpression attenuated miR193b‐induced apoptosis (p < 0.05). Moreover, we found that the Akt pathway negatively regulated miR193b, also affecting apoptosis. Further analyses indicated that PIK3CA mutation and KRAS amplification are two mutually exclusive pathways (p < 0.01), and we hypothesize that both two pathways could result in the carcinogenic overactivation of KRAS. Thus, our results suggest that the Akt‐miR193b‐KRAS axis may act as a mechanism affecting apoptosis in gastric cancer cells.

In recent years, microRNA‑193b (miR‑193b) is regarded as a tumor suppressor in the development and progression of various cancers. Several studies have indicated that KRAS could be regulated by miR‑193b in pancreatic cancer cells. However, the function of miR‑193b in human esophageal squamous cell carcinoma has not been explored intensively thus far. Herein, the relationship between miR‑193b and KRAS was mainly explored in esophageal squamous cell carcinoma cells. In the present study, the expression levels of miR‑193b and KRAS were assessed in both human esophageal cancer cells and tissues. The direct regulatory relationship between miR‑193b and KRAS was evaluated using dual‑luciferase assay. The effect of miR‑193b overexpression and inhibitor on cell proliferation, migration/invasion, and apoptosis was further detected herein. Our results indicated that the expression of miR‑193b was significantly lower in human esophageal cancer tissues than paracancerous tissues. The expression level of miR‑193b/KRAS was stage‑dependent in human esophageal cancers. KRAS was indicated as the direct target of miR‑193b, and upregulation of miR‑193b increased the percentage of cell apoptosis, and suppressed cell proliferation as well as cell migration/invasion via direct regulation of KRAS. Therefore, our study indicated that miR‑193b plays an important role in the development and progression of human esophageal squamous cell carcinoma, which may become a novel target in the treatment of human esophageal squamous cell carcinoma in the future.