Annals of clinical and laboratory science最新文献

Objective: To elucidate the molecular mechanisms underlying ischemia-reperfusion (I/R) injury and explore novel therapeutic targets.

Methods: Raw counts from control, middle cerebral artery occlusion (MCAO), and MCAO+drug groups were processed via the Differential Expression analysis for Sequence data, version 2 (DESeq2) pipeline. Significant differentially expressed genes (DEGs) were identified under thresholds of padj<0.05 and |log2FoldChange|≥1. Functional enrichment analyses, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were conducted using clusterProfiler. A protein-protein interaction (PPI) network of the DEGs was then constructed through Search Tool for Retrieval of Interacting Genes/Proteins (STRING) and visualized in Cytoscape, where Hub genes were screened using CytoHubba. Finally, Hub genes were mapped to the Drug-Gene Interaction Database (DGIdb) to identify potential druggable targets.

Results: A total of 948 upregulated and 419 downregulated DEGs were identified when comparing the MCAO group against controls, while 348 upregulated and 593 downregulated DEGs emerged from the MCAO+drug vs. MCAO comparison. GO and KEGG enrichment analyses indicated that inflammation, immune response, cell cycle, and apoptosis pathways were significantly affected. The PPI network highlighted Cyclin-Dependent Kinase 1 (CDK1), Aurora Kinase B (AURKB), Baculoviral IAP Repeat Containing 5 (BIRC5), and other cell cycle regulators as key Hub genes. DGIdb queries revealed multiple existing inhibitors targeting these Hub genes, suggesting potential therapeutic agents for mitigating cerebral I/R injury.

Conclusion: Our results provide a comprehensive molecular view of cerebral ischemia-reperfusion damage, pinpointing pivotal genes and pathways involved in inflammation and cell cycle regulation. These findings could aid in developing targeted therapies and offer new insights into stroke management. Further experimental and clinical studies are warranted to confirm the efficacy and safety of these potential interventions.

Objective: By establishing an in vitro model of septic cardiomyopathy (SIC), we aimed to investigate whether fasudil exerts its cardioprotective effects by inducing autophagy in cardiomyocytes.

Methods: Rat cardiomyocyte H9c2 cells were cultured in vitro, and lipopolysaccharide (LPS) was added to induce inflammation, simulating an in vitro model of SIC. The cells were divided into five groups, including control group, LPS group, fasudil group, intervention group (fasudil+LPS) and hydroxychloroquine intervention group (fasudil+LPS+hydroxychloroquine). Cell apoptosis was measured using Hoechst 33342/PI fluorescence staining. Changes in mitochondrial membrane potential were measured using JC-1 fluorescence staining. Autophagy and autophagic flux were evaluated using MDC staining and mCherry-EGFP-LC3B assays. Western blotting was conducted to measure the expression of apoptosis-related proteins (Bcl2, Bax, and Cleaved-Caspase3), autophagy-related proteins (LC3B and p62), and the phosphorylation levels of the Akt protein.

Results: In the LPS group, H9c2 cells showed significant apoptosis and decreased mitochondrial membrane potential, which were markedly alleviated after treatment with fasudil. Western blotting revealed increased expression of the pro-apoptotic protein Bax in the LPS group, and treatment with fasudil inhibited LPS-induced up-regulation of Bax expression (P<0.05). Autophagic flux analysis showed that fasudil increased autophagic activity and reduced LC3B accumulation. Western blotting indicated that fasudil up-regulated the protein levels of the autophagy marker LC3B (P<0.05), inducing autophagy in H9c2 cells. However, the protein levels of both LC3B and the autophagosome adaptor protein p62 were significantly down-regulated in the intervention group (P<0.01), suggesting that fasudil promotes autophagic flux in H9c2 cells. Hydroxychloroquine potently inhibited autolysosome-dependent degradation of LC3B and p62 proteins, resulting in their accumulation. The phosphorylation level of the Akt protein was markedly up-regulated in the LPS group but was significantly down-regulated in the fasudil group. An even greater decrease in Akt phosphorylation was observed in the intervention group (P<0.01).

Conclusion: Fasudil induces autophagy in H9c2 cells. Treatment with LPS promotes autophagic flux and inhibits apoptosis, thereby exerting protective effects on the cells.

Objective: Anemia of inflammation is common in patients with infections, autoimmune diseases, chronic inflammation, and various cancers. Few studies have investigated the effects of apoptosis, lipocalin-2, and cytokines on erythropoiesis in anemia of inflammation. This study explored the relationship between serum-soluble apoptotic biomarkers, lipocalin-2, erythropoietin, inflammatory cytokines, and reticulocyte production in anemia of inflammation.

Methods: A total of 164 patients with systemic inflammation were evaluated. Levels of APO-1, lipocalin-2, erythropoietin, tumor necrosis factor (TNF)-α, TNF-related apoptosis-inducing ligand (TRAIL), and interleukin (IL)-6 were measured. To assess erythropoietic activity, the reticulocyte maturity index (RMI) was calculated using subfractions of immature reticulocytes.

Results: Patients with elevated levels of APO-1, lipocalin-2, and TNF-α had significantly lower RMI than those without elevations in these parameters. Compared with non-anemic individuals, anemic patients had significantly higher levels of APO-1, TNF-α, and lipocalin-2. Lipocalin-2 was negatively correlated with RMI and hemoglobin levels and positively correlated with APO-1, TNF-α, and IL-6 levels but not with erythropoietin and TRAIL levels. Patients in the highest quartile of lipocalin-2 levels exhibited significantly higher APO-1 levels than those in the lowest quartile of lipocalin-2 levels. Elevation of lipocalin-2 resulted in a 1.3-fold increase in the prevalence of low RMI (odds ratio: 1.314, 95% confidence interval, 1.011-2.612, p<0.001).

Conclusion: APO-1 and lipocalin-2 are associated with the development of anemia in inflammatory conditions, possibly by inhibiting reticulocyte production, particularly in conjunction with inflammatory cytokines.

Objective: The chromosomal abnormality (CA) in newly diagnosed multiple myeloma (NDMM) suggests the involvement in disease initiation and progression. Fluorescence in situ hybridization (FISH) provides high-sensitivity detection of these aberrations. To elucidate the clinicopathological impact of CAs, we integrated bone marrow (BM) morphology, FISH, flow cytometry (FCM), and PET/CT to analyze their interrelationships in a cohort of 156 NDMM patients.

Methods: BM morphology, FISH, FCM, and PET/CT were performed in 156 patients with NDMM. MM-related serum and urinary markers were collected.

Results: Gain or amplification of 1q (gain/amp(1q)), del(13q), del(17p), and immunoglobulin heavy chain (IGH) translocation were observed in 43.59%, 31.41%, 7.69%, and 25% of NDMM patients, respectively. 16 NDMM patients had extramedullary disease (EMD). The SUVmax median was 5.05 (3.93-7.4). Median BM plasma cell ratio and abnormal plasma cell ratio in NDMM patients were 22.25% and 6.475%, respectively. The above cell ratios were inversely correlated with IgA and IgM, positively correlated with creatinine, serum, and urinary β-2 microglobulin, and were proved lower in NDMM patients without CAs. In IgG type NDMM, M protein was significantly associated with the two ratios. Abnormal plasma cell ratio was lower in CD56 negative group and BM plasma cell ratio was lower in R-ISS I stage group.

Conclusions: The NDMM patients with CAs had higher BM plasma cell ratio, serum and urinary β-2 microglobulin, 24-hour urinary protein, and abnormal plasma cell ratio than the NDMM patients without CAs. The application of BM morphology, FISH, and FCM has significance for the prognosis of NDMM.

Non-sebaceous lymphadenoma (NSL) of the parotid gland is a rare benign tumor. During diagnosis, NSL may be confused with malignant tumors such as lymphoepithelial carcinoma, which can result in overtreatment. In this article, the clinical data and histopathological and immunohistochemical characteristics of two NSL patients were retrospectively analyzed, and the relevant literature was reviewed. The clinical primary manifestation of the cases was a painless mass located in the lower pole of the parotid gland. On histological examination, the tumor was found to comprise epithelial components and lymphoid stroma. The epithelial components formed a cystic cavity, a solid island, or a duct-like structure. The cystic cavity and duct-like structure were composed of two types of cells: luminal cells and outer cells. Within the stroma, proliferating lymphoid follicles were present with well-defined boundaries from the surrounding parotid gland tissue. Immunohistochemical analysis showed that epithelial cells were positive for CK5/6, luminal cells were positive for CK7, and outer cells were positive for p63 and p40. NSL is a rare benign tumor. There are high demands placed on pathologists for accurate intraoperative frozen section diagnoses, which can be challenging. This summary of two NSL cases and a literature review may aid in the clinical recognition of NSL and improve the accurate diagnosis of this benign tumor.

Objective: This study unveils how obacunone (OB) mitigates ulcerative colitis (UC)-induced colonic epithelial cell injury by activating mitophagy via the poly(ADP-ribose) polymerase 1 (PARP1)/sirtuin 1 (SIRT1) signaling pathway.

Methods: Bioinformatics analyses and network pharmacology were utilized to identify potential OB-related targets in UC, after which Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted. Core targets were identified via Cytoscape network modeling and machine learning algorithms. The dextran sulfate sodium (DSS)-treated human intestinal epithelium cell line (NCM460) served as an in vitro model of UC. Prior to DSS treatment, cells were pre-incubated with OB and/or the mitophagy inhibitor Mdivi-1. Furthermore, PARP1-overexpressing NCM460 cells were co-treated with OB and DSS. Cell viability and apoptosis, as well as mitochondrial reactive oxygen species (mtROS) levels were measured using the CCK-8 assay, flow cytometry, and MitoSOX Red staining, respectively. The expression of PARP1, SIRT1, PTEN-induced putative kinase 1 (PINK1), Parkin, and microtubule-associated protein 1A/1B-light chain 3 (LC3) was evaluated through Western blot analysis.

Results: 145 potential targets implicated in bacterial component recognition, oxidative stress response, and inflammation/apoptosis regulation were identified after bioinformatics and network pharmacology analyses. Machine learning algorithms further refined these to five core targets (PARP1, HIF1A, PTGS2, ITGB1, and HSP90AB1), all of which demonstrated strong binding affinity with OB. In DSS-treated NCM460 cells, OB significantly improved cell viability, while reducing apoptosis and mtROS production. Mechanistic studies revealed that OB suppressed PARP1 expression while upregulating SIRT1, PINK1, Parkin, and LC3II. Notably, the protective effects of OB were abrogated by Mdivi-1 treatment or PARP1 overexpression, both of which decreased cell viability, increased apoptosis and mtROS levels, and downregulated PINK1, Parkin, and LC3II expression.

Conclusion: OB attenuates UC-associated epithelial damage through PARP1/SIRT1-dependent mitophagy activation.

Objective: To evaluate the analytical performance of the Maglumi® X6 ABEI-based automated immunoassay for quantifying PIVKA-II, a clinically relevant biomarker for hepatocellular carcinoma (HCC), and to compare its performance with the Lumipulse® assay.

Methods: Imprecision was assessed following CLSI EP15-A3 guidelines using internal quality controls (IQCs) at low and high concentrations, measured three times daily for five days. Method comparison was performed on 63 patient serum samples, and reference values were assessed in 25 healthy individuals.

Results: Within-run coefficients of variation (CVs) for the Maglumi® assay were 17.19% and 3.65% at 56 and 5026 mAU/mL, respectively. Between-run CVs were 1.47% and 2.14% for the same IQCs. Accuracy showed biases of 13.51% and 0.53% at low and high IQC, respectively. Comparison with Lumipulse® showed strong correlation (r=0.95) with a mean bias of 49.9 mAU/mL. Among healthy subjects, the 95th percentile was 45 mAU/mL.

Conclusion: The Maglumi® X6 ABEI-based immunoassay demonstrated good analytical performance. Further studies with larger cohorts are warranted to confirm clinical performance.

Objective: This study aims to investigate the regulatory effects of cigarette smoke extract (CSE) on resistin-like molecule β (RELMβ) in chronic obstructive pulmonary disease (COPD) and to elucidate the mechanisms by which RELMβ modulates inflammatory factors.

Methods: The expression of RELMβ in 16HBE cells after CSE exposure was detected. The activation and nuclear translocation of NF-κB/p65, degradation of IκBα protein, and the expression of inflammatory factors were assessed in 16HBE cells with RELMβ overexpression or exogenous human recombinant RELMβ protein stimulation.

Results: CSE exposure promoted mRNA and protein expression of RELMβ in 16HBE cells. Both stable overexpression of RELMβ and exogenous recombinant RELMβ protein stimulation enhanced secretion of IL-1β, IL-8, IL-6, and TNF-α, upregulated mRNA expression of IL-1β and IL-8, increased phosphorylation of p-IKKα/β, p-IκBα, and p-NF-κB/p65, promoted nuclear translocation of NF-κB/p65, and accelerated the degradation of IκBα in a dose-dependent and time-dependent manner. Inhibition of the NF-κB signaling pathway using pyrrolidine dithiocarbamate and Bay 11-7082 blocked the promotion effect of CSE and RELMβ on expression of p-NF-κB/p65, IL-1β, and IL-8.

Conclusion: CSE induces inflammation via RELMβ through NF-κB/p65 signaling pathway in COPD. This study provides new insights into the involvement of CSE in the progression of COPD from the perspective of RELMβ.