Current Molecular Medicine (Volume 26 - Issue 4)

Hemolysis Analysis and Hemolysis-related MicroRNA Candidates for Serum/Plasma Samples

2026-04-06

Hemolysis is a major challenge in the screening and validation of serum or plasma miRNA biomarkers for human diseases. Over the past decade, numerous studies have focused on hemolysis detection at both the pre-analytical and postanalytical stages to minimize bias in miRNA quantification. Both conventional and advanced hemolysis determination methods have played important roles in quality control in hemolysis assessment and risk prediction during the plasma or serum miRNA quantification process. This review discusses the advantages of these methods and provides an interactive summary of prior knowledge on hemolysissensitive miRNAs and their potential applications in disease diagnosis. Furthermore, the review highlights the advancements in machine learning technologies that enhance classifier predictions and hemolysis risk model evaluations, particularly during the post-analytical stage. Finally, it discusses the ongoing development, standardization, and potential applications of these approaches, which will contribute to a more comprehensive and interpretable framework for the discovery and validation of plasma or serum miRNA biomarkers.

SMAC/DIABLO: A Guardian Angel in Boosting Anticancer Drug-Induced Apoptosis

2026-04-06

Apoptosis is an established hallmark of cancer. In normal conditions, apoptosis is strictly controlled; however, when it is not properly managed, it causes several complications, including cancer progression and drug resistance. SMAC/ Diablo (SMAC) is a mitochondrial protein that is released into the cytosol upon activation of BAX/BAK channels with apoptotic signals. SMAC protein interacts and neutralizes inhibitors of apoptosis (IAP) proteins and initiates the caspase cascade, which leads to apoptosis. SMAC is downregulated in several types of cancer, which led to the design of small-molecule inhibitors known as SMAC mimetics as new cancer therapeutics, and some of these molecules are in the clinical phase. It has also been shown that a combination of SMAC with standard anti-cancer drugs could be beneficial to drug-resistant cancer. Despite being a pro-apoptotic protein, it has been found that SMAC/Diablo is overexpressed in several types of cancers like lung, breast, bladder, cervix, pancreas, prostate, and colon, as well as in melanoma and glioma, and in cancer cells. Recently, we have reported that the overexpression of SMAC in cancers is essential for cell and tumor growth due to non-apoptotic regulation of phospholipid synthesis. The current review is focused on apoptotic and non-apoptotic functions of SMAC and its role in drug resistance.

Exercise Alleviates Atherosclerosis Through the Modulation of the NLRP3 Inflammasome

2026-04-06

Atherosclerosis (AS) is a chronic inflammatory disease closely associated with endothelial dysfunction and oxidative stress. The NOD-like receptor protein 3 (NLRP3) inflammasome, a key regulator of inflammatory responses, can exacerbate the progression of AS when activated. Growing evidence suggests that exercise, as a non-pharmacological intervention, can alleviate the progression of AS by modulating the activity of NLRP3 inflammasome. This review discusses how exercise influences the development of AS through the regulation of NLRP3 inflammasome and the underlying molecular mechanism. This study introduces the structure and activation mechanisms of NLRP3 inflammasome, as well as its role in AS. And summarizes how exercise can ameliorate endothelial dysfunction, regulate lipid metabolism, and suppress oxidative stress and inflammation by affecting the expression and activity of NLRP3 inflammasome, thereby exerting a beneficial impact on AS. Additionally, we explore the effects of exercise on the downstream inflammatory cytokines of NLRP3 inflammasome and how this regulation could help to slow the progression of AS. These findings underscore the therapeutic relevance of exercise in the prevention and treatment of AS. It provides new insights into the role of exercise interventions in the management of AS and lays a theoretical foundation for the development of innovative treatment strategies for cardiovascular disease. Given that the NLRP3 inflammatome plays an important role in the pathogenesis and treatment of AS, exercise therapy strategies targeting the NLRP3 inflammatome will help promote the development of precision medicine for AS.

Dysregulation of Key Biological Processes in Endometriosis Pathophysiology

2026-04-06

Endometriosis is a typical disorder affecting the female reproductive system and is characterized by the presence of tissue resembling the endometrium both within and beyond the pelvic cavity. Unfortunately, the etiology of endometriosis is not well understood. The purpose of this document is to create a summary of the factors contributing to endometriosis, especially the dysregulation of cellular and molecular pathways. Key biological processes implicated include enzyme dysregulation, exosome dysfunction, hormonal imbalances, apoptosis, angiogenesis, oxidative stress, epigenetic dysregulation, and the involvement of cytokines and chemokines. Many of these factors have overlapping pathways that can enhance the survival of endometrial debris and facilitate the implantation of endometrial tissue in extrauterine sites. This knowledge can provide a broader perspective on the onset and progression of endometriosis. Additionally, this study paves the way for the discovery of new therapeutic targets to improve the efficacy of endometriosis treatments and reduce the side effects associated with current treatments. Further research is needed to better understand the underlying mechanisms that lead to the dysregulation of diseaserelated pathways, which could ultimately be useful in early diagnosis and disease staging.

CD82 Methylation Patterns in Inflammatory Arthritis and their Clinical Association with Hypertension in Rheumatoid Arthritis

2026-04-06

Objective: This study aimed to compare CD82 methylation patterns in peripheral blood among patients with rheumatoid arthritis [RA], inflammatory arthritis, and healthy controls [HC] and to assess their clinical associations with hypertension in RA.

Methods: In this cross-sectional study, CD82 methylation at positions 44596705- 44596865 on chromosome 11 was analyzed using targeted methylation techniques in peripheral blood from patients with RA, psoriatic arthritis [PsA], ankylosing spondylitis [AS], gout, and HC.

Results: CD82 cg22143324 methylation levels were significantly different between RA patients and healthy controls [P<0.0001], PsA [P=0.0281], and AS [P=0.0360]. In RA subgroups, individuals negative for both rheumatoid factor [RF] and cyclic citrullinated peptide [CCP] [RA-DN], as well as those positive for both [RA-DP], exhibited significantly different methylation levels compared to HC [P=0.0355 and P<0.0001, respectively]. ROC analysis indicated a promising diagnostic potential for CD82 cg22143324 methylation, especially with the TTT haplotype. Correlation analysis revealed significant associations between CD82 methylation and CCP levels, as well as hypertension in RA patients.

Conclusion: The analysis conducted revealed altered CD82 cg22143324 methylation in RA, with potential utility in distinguishing seronegative patients from healthy controls. An association between lower methylation levels and comorbid hypertension in RA patients was also observed, warranting further investigation.

Non-coding RNAs PROX1-AS1 and miR-647: Potential Interaction and Prognostic Value in Gastric Cancer

2026-04-06

Background: Gastric cancer (GC) remains one of the most common malignancies and the third cause of cancer-related deaths worldwide. Non-coding RNAs (ncRNAs), including microRNAs and long ncRNAs, can contribute to the pathogenesis and progression of GC and therefore could be its potent diagnostic and prognostic biomarkers. The aim of our work was to estimate the expression of PROX1- AS1 (Prospero Homeobox 1 Antisense RNA 1) and miR-647 (microRNA-647) in GC and investigate their potential interaction and clinical significance.

Methods: The study included tumor and adjacent non-tumor tissues from 110 GC patients and plasma samples from 65 GC patients; 38 sectional normal gastric tissue samples and 49 plasma samples of healthy donors were included as controls. Expression levels of both ncRNAs were quantified in all samples by using real-time polymerase chain reaction (RT-PCR) and their possible correlations with the clinical and pathological characteristics of patients were analyzed. A potential inverse correlation between PROХ1-AS1 and miR-647 expression was addressed by in vitro experiments in a panel of cancer cell lines.

Results: The expression of PROX1-AS1 and miR-647 was not significantly different in tissues of GC patients and sectional normal gastric tissue samples. However, they have demonstrated a negative correlation both in the tumor and the adjacent nontumor tissue of GC patients. PROX1-AS1 expression was significantly decreased in GC tissues, whereas the miR-647 expression was increased. The expression of the ncRNAs was associated with clinical and pathological characteristics of GC patients. The overexpression of miR-647 led to a significant decrease in PROX1-AS1 expression in five cancer cell lines, including the GC cell line SNU-1.

Conclusion: We have demonstrated a negative correlation between PROX1-AS1 and miR-647 in both GC tissues and the cancer cell lines. In addition, expression of both ncRNAs was associated with the primary tumor size. Therefore, these ncRNAs might have potential prognostic value.

Ginsenoside Rg1 Attenuates Muscle Atrophy in Hyperglycemic Conditions, Inactivity and Protein Deprivation Models via AKT/mTOR Pathway Activation

2026-04-06

Background: Muscle atrophy, a debilitating condition prevalent in diabetes and extended periods of immobilization, lacks robust therapeutic strategies. This investigation examines ginsenoside Rg1's therapeutic potential in counteracting muscle atrophy under hyperglycemic conditions and in experimental models of immobilization and dietary protein restriction.

Methods: C2C12 murine myoblasts were cultured under variable glucose concentrations and treated with or without Rg1. Multiple cellular parameters were evaluated, including cell viability, apoptotic indices, cell cycle distribution, and protein synthesis rates. The activation status of the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling cascade and expression of atrophy-related markers were quantified using qRT-PCR and Western blot analyses. In parallel animal studies, rats were subjected to either immobilization or protein restriction protocols, with or without Rg1 administration. Muscle function, mass, and relevant biomarkers were evaluated.

Results: Hyperglycemic conditions significantly compromised C2C12 myoblast viability, triggered apoptotic pathways, and disrupted normal cell cycle progression. Rg1 administration effectively attenuated these detrimental effects through enhanced AKT/mTOR pathway activation, upregulation of Myogenin (MyoG) expression, and suppression of atrophy-associated markers. In the rat models, Rg1 supplementation significantly ameliorated muscle deterioration, maintaining muscle mass, contractile force, and exercise tolerance, while simultaneously modulating atrophy signaling pathways and attenuating inflammatory responses. The protective effects of Rg1 were abrogated after the co-treatment with an AKT inhibitor.

Conclusion: Ginsenoside Rg1 exhibits significant protective properties against muscle atrophy under hyperglycemic conditions and in experimental models of immobilization and protein restriction, primarily mediated through activation of the AKT/mTOR signaling pathway. These findings establish Rg1 as a promising therapeutic candidate for the treatment of muscle atrophy.

Pancreatic Elastase Affects Liver Injury by Activating Proinflammatory Cytokines in Kupffer Cells via the JAK2/STAT3 Signaling Pathway

2026-04-06

Introduction: This study aimed to investigate the role of JAK2 (Janus kinase 2)/STAT3 (signal transducer and activator of transcription 3) signaling in liver injury during severe acute pancreatitis (SAP), focusing on pancreatic elastase- and lipopolysaccharide (LPS)-induced Kupffer cell (KC) activation.

Methods: A rat SAP model was established via retrograde taurocholic acid infusion into the biliopancreatic duct. Inflammatory cytokine levels and JAK2/STAT3 pathway activity were quantified in liver tissues. KCs were treated with elastase/LPS ± AG490 (JAK2 inhibitor). Proinflammatory cytokines, RNA, and protein expression were analyzed.

Results and Discussion: SAP rats exhibited elevated TNF-α, IL-6, and IL-18 levels in both serum and liver tissues, with JAK2/STAT3 pathway activation. AG490 administration suppressed JAK2/STAT3 activation, reduced inflammation, and alleviated liver injury. Similarly, KCs treated with elastase and LPS showed increased proinflammatory cytokine levels and JAK2/STAT3 upregulation, which were mitigated by AG490 treatment.

Conclusion: The findings highlighted the pivotal role of the JAK2/STAT3 signaling pathway in SAP-induced liver injury. Selective inhibition of this pathway by AG490 could reduce inflammation and protect against liver damage, suggesting its potential as a therapeutic target for inflammatory liver diseases.

Causal Relationship Between Plasma Protein-to-Protein Ratios and Rheumatoid Arthritis: A Proteome-wide Mendelian Randomization Study

2026-04-06

Introduction: The causal relevance of circulating plasma protein-to-protein ratios (PPRs) in Rheumatoid Arthritis (RA) remains unclear. We employed Mendelian Randomization (MR) to investigate this relationship.

Methods: This study utilized summary data of ratio quantitative trait loci (rQTLs) for 2,821 circulating PPRs from the GWAS Catalog and two RA-related GWAS datasets (FinnGen and GWAS Catalog). Causal estimates were obtained using various Mendelian randomization (MR) methods, including IVW and MR-Egger regression. Significant PPRs were further analyzed via protein–protein interaction (PPI), functional enrichment, and druggability assessments. Key genes were validated using qPCR.

Results: Fifteen candidate PPRs with consistent directional effects, and nine core PPRs, achieved statistical significance in both datasets. Protein–protein interaction (PPI) network analysis revealed involvement of these proteins in various biological processes. Gene Ontology (GO) analysis indicated roles in immune response and protein binding, while KEGG pathway analysis showed enrichment in Toll-like receptor signaling pathways. Friends analysis identified UBAC1 as a key gene, and seven PPR-associated proteins were found to be druggable. qPCR validation confirmed differential expression of UBAC1, CD40, ITGB5, and GLOD4.

Discussion: Our findings establish a robust genetic causal link between specific PPRs and RA, moving beyond association to suggest potential etiology. Integrated analyses prioritize UBAC1, CD40, ITGB5, and GLOD4 as key contributors to RA pathogenesis, with functional enrichment indicating their involvement in immune and inflammatory pathways. The druggability of several implicated proteins underscores the translational potential of these results.

Conclusions: This study used MR to establish a causal relationship between plasma PPRs and RA risk. UBAC1, CD40, ITGB5, and GLOD4 may play key roles in the pathogenesis of RA.