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Biomedicines

Biomedicines is an international, peer-reviewed, open access journal on biomedicines published monthly online by MDPI.

Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
High Visibility: indexed within Scopus, SCIE (Web of Science), PubMed, PMC, CAPlus / SciFinder, and other databases.
Journal Rank: JCR - Q1 (Pharmacology and Pharmacy) / CiteScore - Q1 (Medicine (miscellaneous))
Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 14.6 days after submission; acceptance to publication is undertaken in 2.8 days (median values for papers published in this journal in the second half of 2024).
Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.
Companion journals for Biomedicines include: IJTM, BioMed, Anesthesia Research and Emergency Care and Medicine.

Impact Factor: 3.9 (2024); 5-Year Impact Factor: 4.2 (2024)

Imprint Information Journal Flyer Open Access ISSN: 2227-9059

Latest Articles

12 pages, 1336 KiB

Open AccessArticle

Evaluating Genomic and Clinical Risk Factors for Alzheimer’s Disease in Individuals with Hypertension

by Elizabeth Kim, Kevin Zhang, Miski Abdi, Wei Tse Li, Ruomin Xin, Jessica Wang-Rodriguez and Weg M. Ongkeko

Biomedicines 2025, 13(6), 1508; https://doi.org/10.3390/biomedicines13061508 - 19 Jun 2025

Background/Objectives: Alzheimer’s disease (AD) is a progressive neurodegenerative condition whose growing prevalence has become an increasingly important public health concern as the population ages. The lack of a definitive cure elevates the importance of identifying risk factors that are crucial for prevention efforts. [...] Read more.

Background/Objectives: Alzheimer’s disease (AD) is a progressive neurodegenerative condition whose growing prevalence has become an increasingly important public health concern as the population ages. The lack of a definitive cure elevates the importance of identifying risk factors that are crucial for prevention efforts. Hypertension (HTN) and obesity have emerged as two highly widespread, interrelated conditions that have independently been associated with AD risk. Despite extensive research into AD pathology, the impact of obesity in a hypertensive population is not well explored. This study aims to investigate how obesity and blood pressure control within a hypertensive population may interact with genomic risk and environmental factors to influence AD incidence. Methods: A retrospective cohort of matched AD and normal patients diagnosed with HTN and taking anti-HTN drugs (n = 1862) from the All of Us database was analyzed. In this hypertensive cohort, obesity was significantly associated with increased AD risk. Genome-wide association studies (GWASs) were conducted on hypertensive AD individuals (n = 1030) and identified six single nucleotide variants (SNVs) that were associated with AD development in this population. Results: Obesity and Area Deprivation Index, a measure of socioeconomic status, were significantly associated with elevated AD risk within the hypertensive cohort. GWAS analysis identified six SNVs significantly associated with AD development among the hypertensive cohort. Conclusions: Our findings suggest that among hypertensive individuals, comorbid obesity and the Area Deprivation Index confer greater AD risk. These results highlight the critical need for obesity prevention and management strategies as part of Alzheimer’s risk reduction efforts. Full article

(This article belongs to the Section Neurobiology and Clinical Neuroscience)

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25 pages, 540 KiB

Open AccessReview

Malignancies in Celiac Disease—A Hidden Threat with Diagnostic Pitfalls

by Aleksandra Kubas and Ewa Małecka-Wojciesko

Biomedicines 2025, 13(6), 1507; https://doi.org/10.3390/biomedicines13061507 - 19 Jun 2025

Celiac disease (CeD) is an autoimmune disorder that is triggered by gluten ingestion in genetically predisposed individuals. Untreated or poorly controlled CeD leads to various disease complications, such as malnutrition, osteoporosis, autoimmune diseases, or refractory celiac disease (RCD). Accumulating recent research has highlighted [...] Read more.

Celiac disease (CeD) is an autoimmune disorder that is triggered by gluten ingestion in genetically predisposed individuals. Untreated or poorly controlled CeD leads to various disease complications, such as malnutrition, osteoporosis, autoimmune diseases, or refractory celiac disease (RCD). Accumulating recent research has highlighted the association between CeD and the development of malignancies, particularly enteropathy-associated T-cell lymphoma (EATL) and small bowel carcinoma (SBC), which are neoplasms with extremely poor prognoses. Genetic alterations in the JAK1–STAT3 pathway and the high prevalence of microsatellite instability may be the main drivers of CeD-associated lymphomagenesis and small bowel oncogenesis and therefore could be an attractive therapeutic target to block cancer transformation. However, to date, the risk factors and exact mechanisms underlying malignancy development in patients with CeD remain unclear, and prospective cohort studies that include molecular profiling are needed. Moreover, current guidelines on the management of CeD do not provide standardized protocols for cancer surveillance—particularly regarding screening intervals, risk stratification, and monitoring strategies for high-risk patients such as those with RCD. This paper reviews the existing knowledge on malignancies in CeD, highlights diagnostic challenges, and discusses future perspectives on the early detection, monitoring, and treatment of CeD-associated neoplasms. Full article

(This article belongs to the Section Cancer Biology and Oncology)

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21 pages, 3504 KiB

Open AccessArticle

Genotype-Based Housing as a Potential Confounder in Studies Using Transgenic Mouse Models—Insight from the A53T Mouse Model of Parkinson’s Disease

by Olga Dubljević, Miodrag Dragoj, Milica Potrebić Stefanović, Maja Srbovan, Miloš Stanojlović and Željko Pavković

Biomedicines 2025, 13(6), 1506; https://doi.org/10.3390/biomedicines13061506 - 19 Jun 2025

Background/Objectives: Environmental factors, including the differences in genotype-based housing (GbH), can act as confounding variables in studies using transgenic mouse models, potentially influencing experimental outcomes and limiting their reproducibility and translational value. Despite the widespread use of transgenic models in preclinical studies, [...] Read more.

Background/Objectives: Environmental factors, including the differences in genotype-based housing (GbH), can act as confounding variables in studies using transgenic mouse models, potentially influencing experimental outcomes and limiting their reproducibility and translational value. Despite the widespread use of transgenic models in preclinical studies, the extent to which housing conditions can affect the behavioral and molecular parameters of interest remains poorly understood. This study aims to investigate how different GbH conditions influence visuo-spatial memory and gene expression in the A53T mouse model (JAX006823) of Parkinson’s disease (PD) during the pre-motor phase. Methods: A53T+ transgenic male mice and their non-transgenic littermates (A53T−) were housed in either mixed-genotype (MGH) or single-genotype (SGH) environments from postnatal day (PND) 30, with C57BL/6J mice serving as the controls. A behavioral assessment using the Novel Object Recognition and Object Location Tests was conducted at PND 180, followed by a qPCR analysis of Iba1, Gfapα, Bdnf, Tnfα, Il-1β, and Il-6 expression in the medial prefrontal cortex and the hippocampus. Results: The variations in GbH influenced behavior and mRNA expression differently in the A53T+ and A53T− animals. Specifically, the A53T− mice in SGH environments displayed behavioral and molecular profiles similar to the C57BL/6J controls, while the same was not evident in the MGH environments. In the A53T+ mice, the mRNA expression of Iba1, Gfapα, Bdnf, and Tnfα was sensitive to variations in GbH, while memory impairment was not. Conclusions: This study highlights the importance of considering environmental factors in studies using transgenic animal models. The obtained data suggests that GbH can influence the parameters of interest in preclinical research, implicating the need for the optimization of future study designs. Full article

(This article belongs to the Special Issue Animal Models of Human Pathology: Revision, Relevance and Refinements (3rd Edition))

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15 pages, 1291 KiB

Open AccessSystematic Review

Vaginal Cleansing and Post-Cesarean Infectious Morbidity? Updated Systematic Review and Meta-Analysis of Randomized Trials

by Marco La Verde, Marco Torella, Irene Iavarone, Rossella Molitierno, Antonio Cerillo, Margherita Casillo, Maria Maddalena Marrapodi, Mario Fordellone, Liliana Mariani, Chiara Melito, Barbara Gardella and Mattia Dominoni

Biomedicines 2025, 13(6), 1505; https://doi.org/10.3390/biomedicines13061505 - 19 Jun 2025

Background: Endometritis, maternal fever and wound infection represent the most frequent post-cesarean complications. The aim of the present research was to evaluate the incidence of post-cesarean infections after vaginal cleansing. Materials and methods: The databases analyzed were MEDLINE, Scopus, EMBASE, CENTRAL, [...] Read more.

Background: Endometritis, maternal fever and wound infection represent the most frequent post-cesarean complications. The aim of the present research was to evaluate the incidence of post-cesarean infections after vaginal cleansing. Materials and methods: The databases analyzed were MEDLINE, Scopus, EMBASE, CENTRAL, Google Scholar, Clinicaltrials.gov and the Register of Controlled Trials. No language or geographical restrictions were applied. We included only randomized controlled trials that analyzed various vaginal antiseptic solutions to reduce postpartum endometritis. The terms employed were as follows: vaginal solution, cesarean section, endometritis, wound infection, chlorhexidine, povidone, metronidazole, cetrimide, and pregnancy. The PICO categorization was as follows: P—population: pregnant women; I—intervention: vaginal antiseptic; C—control: hands-off or routine care; O—outcome: post-cesarean endometritis, wound infection and postoperative fever; S—study design: randomized controlled trials. Results: A total of 32 articles, including 13,853 participants, were selected. The vaginal cleansing group showed a low incidence of endometritis. The chlorhexidine group had an OR of 0.56 (95% CI 0.45–0.70, p = 0.010). The povidone group had an OR of 0.47 (95% CI 0.37–0.59, p = 0.002). Considering maternal fever, 2598 patients from 5 studies in the chlorhexidine group were analyzed, alongside 6965 patients from 18 trials in the povidone group. The povidone group presented an Odds ratio of 0.47 (95% CI 0.38–0.57, p = 0.0001). A reduction in wound infection incidence was observed in the povidone group (OR = 0.59, 95% CI = 0.42–0.82, p < 0.05). Conclusions: Vaginal cleansing before cesarean section, particularly with povidone solutions, reduces the incidence of postoperative endometritis and maternal fever. Full article

(This article belongs to the Special Issue High-Risk Pregnancy, Labor and Delivery)

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33 pages, 2546 KiB

Open AccessReview

A Review of the Relationship Between Insulin and Bone Health

by Sivasree Ravindran, Sok Kuan Wong, Nur-Vaizura Mohamad and Kok-Yong Chin

Biomedicines 2025, 13(6), 1504; https://doi.org/10.3390/biomedicines13061504 - 19 Jun 2025

Insulin, a key hormone primarily involved in glucose metabolism, has emerged as a crucial modulator of bone metabolism. Increasing evidence suggests that insulin influences bone health, but its precise mechanism of action remains unestablished. This review explores the intricate relationship between insulin and [...] Read more.

Insulin, a key hormone primarily involved in glucose metabolism, has emerged as a crucial modulator of bone metabolism. Increasing evidence suggests that insulin influences bone health, but its precise mechanism of action remains unestablished. This review explores the intricate relationship between insulin and bone health, as well as elucidating the mechanism of action involved. Animal models of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) demonstrated distinct skeletal alterations, largely attributed to differences in insulin availability and associated metabolic dysfunction. Insulin deficiency in T1DM was associated with the deterioration of trabecular and cortical bone, whereas insulin resistance in T2DM primarily compromised trabecular bone quality. The route, frequency, and duration of insulin administration have been shown to influence bone-related outcomes. Studies involving insulin receptor silencing have suggested that insulin signalling is essential for normal bone development and maintenance. In humans, inconsistent findings on the effects of circulating insulin levels and insulin resistance on bone health were mainly attributed to heterogeneity in age, gender, metabolic status, study designs, population characteristics, and assessment methods. This review also highlights current knowledge gaps and underscores the need for longitudinal studies and mechanistic research. A clearer understanding of the insulin–bone axis may guide the development of targeted strategies to mitigate skeletal complications in individuals with diabetes mellitus. Full article

(This article belongs to the Section Endocrinology and Metabolism Research)

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29 pages, 1900 KiB

Open AccessArticle

MSC1 Cells Suppress Colorectal Cancer Cell Growth via Metabolic Reprogramming, Laminin–Integrin Adhesion Signaling, Oxidative Stress Resistance, and a Tumor-Suppressive Secretome

by Panagiota-Angeliki Galliou, Niti Argyri, Papaioannou Maria, George Koliakos and Nick A. Papanikolaou

Biomedicines 2025, 13(6), 1503; https://doi.org/10.3390/biomedicines13061503 - 19 Jun 2025

Background/Objectives: Mesenchymal stem cells (MSCs) possess immunomodulatory properties, tumor-homing, and low immunogenicity, making them attractive for cell-based cancer therapies, but their role in colorectal cancer (CRC) remains controversial. The MSC1 phenotype, a pro-inflammatory, tumor-suppressive state induced by short-term, low-dose LPS activation via TLR4, [...] Read more.

Background/Objectives: Mesenchymal stem cells (MSCs) possess immunomodulatory properties, tumor-homing, and low immunogenicity, making them attractive for cell-based cancer therapies, but their role in colorectal cancer (CRC) remains controversial. The MSC1 phenotype, a pro-inflammatory, tumor-suppressive state induced by short-term, low-dose LPS activation via TLR4, has shown therapeutic promise but remains poorly characterized in CRC. We aimed to elucidate MSC1’s tumor-suppressive mechanisms and validate its activity against CRC cells using an integrated bioinformatics and in vitro approach. Methods: We constructed a high-confidence protein-protein interaction (PPI) network in Wharton’s jelly-derived MSCs (WJ-MSCs) following TLR4 activation to uncover enriched signaling pathways, transcriptional regulators, and secreted factors. Functional and transcriptional enrichment analyses pinpointed key mechanisms. We then co-cultured MSC1 cells with CRC cells to assess effects on proliferation and metabolism. Results: Network analysis revealed six tumor-suppressive mechanisms of MSC1 cells: (i) Metabolic reprogramming via enhanced glucose and lipid uptake, phosphoinositide signaling, and membrane/protein recycling, (ii) Robust antioxidant defenses, including SOS signaling and system xc⁻, (iii) Extracellular matrix stabilization and laminin-111–integrin-mediated adhesion, (iv) Secretome with direct anti-cancer effects, (v) Regulation of survival and cancer-associated fibroblasts (CAFs) formation inhibition through balanced proliferation, apoptosis, and epigenetic signals, (vi) Controlled pro-inflammatory signaling with anti-inflammatory feedback. In vitro, MSC1 cells significantly suppressed CRC cell proliferation and metabolic activity versus controls. Conclusions: This study provides the first mechanistic map of MSC1’s tumor-suppressive functions in CRC, extending beyond immunomodulation to include metabolic competition, ECM stabilization, and anti-cancer secretome activity. These findings establish MSC1 cells as a novel therapeutic strategy for CRC in cell-based cancer therapies. Full article

(This article belongs to the Section Cell Biology and Pathology)

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11 pages, 2775 KiB

Open AccessArticle

Pyridostigmine Mitigates Methotrexate-Induced Liver Fibrosis in Rats: Association with Changes in BMP-9, SIRT1, and Endoglin Expression

by Mehmet Ulusan, Mumin Alper Erdogan, Ozkan Simsek, Hilal Ustundag, Zafer Dogan, Bertug Bekir Ciftci, Mesih Kocamuftuoglu, Imdat Orhan and Oytun Erbas

Biomedicines 2025, 13(6), 1502; https://doi.org/10.3390/biomedicines13061502 - 19 Jun 2025

Background and Objectives: Methotrexate (MTX) is a widely utilised pharmaceutical agent in the treatment of various malignancies and inflammatory diseases. However, its clinical utility is often constrained by its potential for hepatotoxicity. Although pyridostigmine is a well-established reversible acetylcholinesterase inhibitor, its potential therapeutic [...] Read more.

Background and Objectives: Methotrexate (MTX) is a widely utilised pharmaceutical agent in the treatment of various malignancies and inflammatory diseases. However, its clinical utility is often constrained by its potential for hepatotoxicity. Although pyridostigmine is a well-established reversible acetylcholinesterase inhibitor, its potential therapeutic role in preventing hepatic injury remains incompletely defined. The present study aimed to investigate whether pyridostigmine provides protective effects against MTX-triggered liver damage in a rat model. Methods: Thirty-six female Wistar albino rats randomly assigned to three groups: control (n = 12), MTX + saline (n = 12), and MTX + pyridostigmine (n = 12). Hepatotoxicity was induced by a single-dose MTX injection (20 mg/kg), followed by daily oral administration of either pyridostigmine (5 mg/kg) or saline for ten consecutive days. Hepatic function markers, oxidative stress parameters, fibrosis-associated mediators, and histopathological changes were assessed. Results: Pyridostigmine significantly attenuated MTX-induced elevations in plasma alanine aminotransferase (p < 0.05) and cytokeratin-18 levels (p < 0.001), and reduced liver and plasma malondialdehyde (MDA) levels (p < 0.05). Additionally, pyridostigmine treatment resulted in reduced levels of transforming growth factor-beta (p < 0.05), bone morphogenetic protein-9 (p < 0.001), and endoglin levels (p < 0.05), as well as increased sirtuin 1 level (p < 0.05). Histopathological examination revealed that pyridostigmine treatment significantly reduced MTX-induced hepatocyte necrosis, fibrosis, and cellular infiltration. Conclusions: Pyridostigmine exerted hepatoprotective effects against MTX-induced liver injury by attenuating oxidative stress, restoring SIRT1 expression, and suppressing pro-fibrotic signaling. These findings indicate that pyridostigmine may hold therapeutic potential for the prevention of MTX-associated hepatotoxicity. Full article

(This article belongs to the Section Cell Biology and Pathology)

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13 pages, 831 KiB

Open AccessArticle

Analysis of Tacrolimus Clearance in Patients with Kidney Transplants from Romania

by Corina Andreea Rotarescu, Ion Maruntelu, Ion Rotarescu, Alexandra-Elena Constantinescu and Ileana Constantinescu

Biomedicines 2025, 13(6), 1501; https://doi.org/10.3390/biomedicines13061501 - 18 Jun 2025

Background/Objectives:Tacrolimus is a key immunosuppressant in kidney transplantation, but its high interindividual pharmacokinetic variability complicates dosing. This study aimed to develop a population pharmacokinetic model and identify the factors explaining variability to optimize tacrolimus therapy in Romanian kidney transplant recipients. Methods: [...] Read more.

Background/Objectives:Tacrolimus is a key immunosuppressant in kidney transplantation, but its high interindividual pharmacokinetic variability complicates dosing. This study aimed to develop a population pharmacokinetic model and identify the factors explaining variability to optimize tacrolimus therapy in Romanian kidney transplant recipients. Methods: The study included 106 kidney transplant recipients treated at Fundeni Clinical Institute (2022–2024). Tacrolimus blood levels were measured using immunoassays, while gene polymorphisms of CYP3A4, CYP3A5, and ABCB1 were identified by real-time polymerase chain reaction. Results: Patients with CYP3A4*1/*1.001 impact clearance (RSE = 11.8%), while hematocrit was a significant covariate for intercompartmental clearance (RSE = 6.14%). Conclusions: Incorporating CYP3A4*1/*1.001 genotype and hematocrit into dosing strategies can improve therapeutic drug monitoring and personalize immunosuppressive therapy. Full article

(This article belongs to the Special Issue An Update on Transplantation Immunology)

19 pages, 2824 KiB

Open AccessArticle

Regulation of Stemness by NR1D2 in Colorectal Cancer

by Sandra Alonso-García, Paula Sánchez-Uceta, Sara Moreno-SanJuan, Jorge Casado, Jose D. Puentes-Pardo, Huda Khaldy, David Lopez-Pérez, María Sol Zurita-Saavedra, Cristina González-Puga, Angel Carazo and Josefa León

Biomedicines 2025, 13(6), 1500; https://doi.org/10.3390/biomedicines13061500 - 18 Jun 2025

Background: Nuclear Receptor Subfamily 1 Group D Member 2 (NR1D2), a transcription factor that regulates the circadian clock, has been described as an oncogene in colorectal cancer (CRC). In several types of cancer, NR1D2 regulates cancer progression and relapse through cancer stem [...] Read more.

Background: Nuclear Receptor Subfamily 1 Group D Member 2 (NR1D2), a transcription factor that regulates the circadian clock, has been described as an oncogene in colorectal cancer (CRC). In several types of cancer, NR1D2 regulates cancer progression and relapse through cancer stem cells (CSCs), although this aspect has not been studied in CRC. On the other hand, p53 is a tumour suppressor gene that appears mutated in approximately a 50% CRCs. Interestingly, p53 is considered to be a crucial nexus between circadian clock deregulation and cancer. In addition, p53 regulates CSC phenotypes. Methods: We developed an in vitro model in which NR1D2 was silenced in three isogenic cell lines with different p53 status. In addition, we analysed the expression of NR1D2 in a cohort of patients and determined its relationship with the characteristics of patients and tumours. Results: In the in vitro model, NRID2 silencing reduces cell growth and decreases stemness, although only in cells harbouring a wild type p53. In contrast, in cells lacking a functional p53 or harbouring a mutated one, NR1D2 knockout increases cell growth and stemness. In patients, NR1D2 expression correlates with poorly differentiated tumours and high expression of CSCs markers, although only in tumours with a wild type p53, corroborating the results obtained in the in vitro model. Conclusions: Although more research is needed to analyse the mechanism by which NR1D2 regulates stemness in a p53-dependent manner, our results highlight the possibility of using NR1D2 antagonists for treating this type of patient and to develop personalised medicine. Full article

(This article belongs to the Special Issue New Insights in Gastric, Colorectal, and Pancreatic Cancer)

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14 pages, 1167 KiB

Open AccessArticle

Role of Extracellular Vesicles in Chronic Post-Embolic Pulmonary Hypertension: Data from an Experimental Animal Model and Patients

by Elva Mendoza-Zambrano, Verónica Sánchez-López, Belén Gómez-Rodríguez, Inés García-Lunar, Daniel Pereda-Arnau, Luis Jara-Palomares, Teresa Elías-Hernández, Ana García-Álvarez and Remedios Otero-Candelera

Biomedicines 2025, 13(6), 1499; https://doi.org/10.3390/biomedicines13061499 - 18 Jun 2025

Background: The pathogenesis of chronic thromboembolic pulmonary hypertension (CTEPH) involves a multifaceted interplay of factors, including incomplete thrombus resolution, endothelial dysfunction, and vascular remodeling. Recent studies have highlighted the role of extracellular vesicles (EVs) in vascular diseases, suggesting their potential involvement in [...] Read more.

Background: The pathogenesis of chronic thromboembolic pulmonary hypertension (CTEPH) involves a multifaceted interplay of factors, including incomplete thrombus resolution, endothelial dysfunction, and vascular remodeling. Recent studies have highlighted the role of extracellular vesicles (EVs) in vascular diseases, suggesting their potential involvement in CTEPH progression. This study aims to investigate the role of EVs from various cellular sources in the development of CTEPH. Methods: An experimental study was conducted using 11 male three-month-old Large-White pigs. The EVs of endothelial origin (EEVs; CD146+), leukocyte-derived EVs (LEVs; CD45+, CD44+), and consistent with mesenchymal-origin EVs (CD90+, CD105+) were quantified. Measurements were taken at baseline, after the first embolization, and prior to each subsequent weekly embolization. Embolizations were repeated until chronic pulmonary hypertension (PH) was generated. Based on these findings, a clinical case-control study was performed involving nine patients previously diagnosed with CTEPH and 18 patients with pulmonary embolism who did not develop CTEPH after two years of follow-up. Results: The experimental study, consistent with the mesenchymal-origin EVs, exhibited a progressive decrease below baseline levels; LEVs decreased after PH was established, while EEVs remained elevated throughout the study. Subsequently, in the clinical case-control study, CD45+ LEVs emerged as a significant association of CTEPH, with an odds ratio (OR) of 21.25 (95% CI: 1.91–236.00; p = 0.013). Conclusions: Inflammation involving LEVs and EEVs plays a crucial role in sustaining the vascular alterations leading to pulmonary vasculature remodeling in CTEPH. Full article

(This article belongs to the Special Issue Molecular and Translational Research in Cardiovascular Disease)

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19 pages, 2909 KiB

Open AccessArticle

Optimization, Characterization and Pharmacological Validation of the Endotoxin-Induced Acute Pneumonitis Mouse Model

by Emese Ritter, Kitti Hohl, László Kereskai, Ágnes Kemény, Dóra Hargitai, Veronika Szombati, Anikó Perkecz, Eszter Pakai, Andras Garami, Ákos Zsembery, Zsuzsanna Helyes and Kata Csekő

Biomedicines 2025, 13(6), 1498; https://doi.org/10.3390/biomedicines13061498 - 18 Jun 2025

Background/Objectives: In preclinical research of airway inflammation, the endotoxin (lipopolysaccharide: LPS)–induced acute interstitial pneumonitis is the most commonly used mechanism model. However, studies apply different LPS serotypes, doses, administration routes, and reference compounds, making result interpretation challenging and drawing conclusions difficult. Therefore, [...] Read more.

Background/Objectives: In preclinical research of airway inflammation, the endotoxin (lipopolysaccharide: LPS)–induced acute interstitial pneumonitis is the most commonly used mechanism model. However, studies apply different LPS serotypes, doses, administration routes, and reference compounds, making result interpretation challenging and drawing conclusions difficult. Therefore, here we aimed to optimize, characterize, and validate this model with dexamethasone in mice. Methods: Pneumonitis was induced by intratracheal LPS (0.25, 1, 2.5, 5 mg/kg; E. coli O111:B4) in C57BL/6J and NMRI mice; controls received phosphate-buffered saline (PBS). Dexamethasone (5 mg/kg i.p.) was used as a positive control. Respiratory functions were measured by restrained plethysmography 24 h after induction, and core body temperature was monitored. Lungs were excised and weighed, and then myeloperoxidase (MPO) activity and histopathological analysis were performed to assess pulmonary inflammation. Results: LPS-induced significant body weight loss, perivascular pulmonary edema, MPO activity increase, neutrophil infiltration, and respiratory function impairment in a dose-independent manner. However, LPS-induced hypothermia dynamics and duration were dose-dependent. The inhibitory effects of the reference compound dexamethasone were only detectable in the case of the 0.25 mg/kg LPS dose on most inflammatory parameters. These results did not differ substantially between C57BL/6J and NMRI mouse strains. Conclusions: Very low doses of LPS induce characteristic functional and morphological inflammatory alterations in the lung, which do not worsen in response to even 20 times higher doses. Since the effect of pharmacological interventions is likely to be detectable in the case of the 0.25 mg/kg LPS dose, we suggest this protocol for testing novel anti-inflammatory agents. Full article

(This article belongs to the Special Issue Animal Models of Human Pathology: Revision, Relevance and Refinements (3rd Edition))

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13 pages, 528 KiB

Open AccessArticle

Neutrophil–Lymphocyte Ratio in Fibromyalgia and Axial Spondyloarthritis: A Potential Biomarker for Diagnosis and Disease Activity

by Miriam Almirall, Esther Espartal, Xabier Michelena, Carlos Suso-Ribera, Mayte Serrat, Sara Marsal and Alba Erra

Biomedicines 2025, 13(6), 1497; https://doi.org/10.3390/biomedicines13061497 - 18 Jun 2025

Objective: The Neutrophil–Lymphocyte Ratio (NLR) has been proposed as an inflammatory biomarker in several diseases, including Fibromyalgia, with controversial results. The objectives of this study were to: (1) compare NLR values among participants with Fibromyalgia, Axial Spondyloarthritis, and healthy controls; (2) assess [...] Read more.

Objective: The Neutrophil–Lymphocyte Ratio (NLR) has been proposed as an inflammatory biomarker in several diseases, including Fibromyalgia, with controversial results. The objectives of this study were to: (1) compare NLR values among participants with Fibromyalgia, Axial Spondyloarthritis, and healthy controls; (2) assess the relationship between NLR and disease activity; and (3) establish diagnostic and activity cut-off values. Methods: A total of 112 age and gender-matched participants were included in each group. NLR values were compared between groups, correlations with disease activity were analyzed, and cut-off values were calculated using Receiver Operating Characteristic (ROC) curves. Results: The NLR was significantly higher in Fibromyalgia patients compared with healthy controls (1.8 ± 0.5 vs. 1.4 ± 0.2; p < 0.001) and in Axial Spondyloarthritis patients compared with both Fibromyalgia patients (2.1 ± 0.3 vs. 1.8 ± 0.5; p < 0.001) and healthy controls (2.1 ± 0.3 vs. 1.4 ± 0.2; p < 0.001). Within disease groups, the NLR was also significantly higher in patients with severe Fibromyalgia (FIQ ≥ 59) compared with non-severe cases (1.9 ± 0.5 vs. 1.7 ± 0.4; p = 0.008) and in patients with high/very high Axial Spondyloarthritis activity compared with those with low/inactive disease (2.3 ± 0.3 vs. 1.9 ± 0.2; p < 0.001). ROC analysis identified the NLR cut-off values of 1.54 for Fibromyalgia diagnosis, 1.64 for severe disease, 1.61 for Axial Spondyloarthritis diagnosis and 1.95 for high/very high disease activity. Conclusions: The NLR may serve as a cost-effective, rapid, and accessible biomarker for establishing diagnosis and disease activity in Axial Spondyloarthritis and, to a lesser extent, in Fibromyalgia. Further research is needed to validate these findings and explore NLR’s role alongside other inflammatory markers. Full article

(This article belongs to the Special Issue Rheumatology Diseases: Advances in the Understanding of Pathogenesis, Biomarker Research and Treatment Target)

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14 pages, 1248 KiB

Open AccessArticle

Multi-Metal Exposure Profiling in ALS Patients in South Korea via Hair Analysis: A Cross-Sectional Study

by Jae-Kook Yoo, Soon-Hee Kwon, Sul-Hee Yoon, Jeong-Eun Lee, Jong-Un Chun, Je-Hyuk Chung, Sang-Yoon Lee, Jeong-Hwan Lee and Yu-Ra Chae

Biomedicines 2025, 13(6), 1496; https://doi.org/10.3390/biomedicines13061496 - 18 Jun 2025

Objectives: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease with an unclear etiology. This study aimed to assess chronic heavy metal exposure in ALS patients in South Korea by comparing hair concentrations of common (Hg, Pb, Cd) and rare (U, Th, Pt) [...] Read more.

Objectives: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease with an unclear etiology. This study aimed to assess chronic heavy metal exposure in ALS patients in South Korea by comparing hair concentrations of common (Hg, Pb, Cd) and rare (U, Th, Pt) metals with healthy controls. Methods: Hair samples were collected from 66 ALS patients and 70 healthy individuals at Rodem Hospital between 2022 and 2025. Metal concentrations were measured using inductively coupled plasma mass spectrometry (ICP-MS) following standardized washing and digestion protocols. Results: ALS patients showed significantly higher levels of Hg, Pb, Cd, Al, As, and U than controls (p < 0.05). Notably, 40% of ALS patients had Hg levels exceeding 50% of the reference upper limit, compared to only 10% of controls. Elevated levels of uranium and other rare metals were also observed in specific ALS cases. Conclusions: These findings suggest a possible association between heavy metal exposure and ALS in South Korea. Hair analysis may serve as a useful tool for identifying environmental factors contributing to ALS pathogenesis. Full article

(This article belongs to the Special Issue Advanced Molecular Mechanisms and Treatment of Neurological Diseases)

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14 pages, 593 KiB

Open AccessReview

Hashimoto’s Thyroiditis and Female Fertility: Endocrine, Immune, and Microbiota Perspectives in Assisted Reproduction—A Narrative Review

by Emilia Cristina Popa, Laura Maghiar, Teodor Andrei Maghiar, Ilarie Brihan, Laura Monica Georgescu, Bianca Anamaria Toderaș, Liliana Sachelarie and Anca Huniadi

Biomedicines 2025, 13(6), 1495; https://doi.org/10.3390/biomedicines13061495 - 18 Jun 2025

Hashimoto’s thyroiditis is the most prevalent autoimmune thyroid disorder, and it disproportionately affects women of reproductive age. Its impact on fertility and assisted reproductive technologies [ART] has become an area of growing clinical interest. Thyroid autoimmunity can influence female reproductive health through multiple [...] Read more.

Hashimoto’s thyroiditis is the most prevalent autoimmune thyroid disorder, and it disproportionately affects women of reproductive age. Its impact on fertility and assisted reproductive technologies [ART] has become an area of growing clinical interest. Thyroid autoimmunity can influence female reproductive health through multiple interconnected mechanisms, including subtle thyroid hormone imbalances, reduced ovarian reserve, altered endometrial receptivity, and dysregulated immune responses. Subclinical hypothyroidism and the presence of anti-thyroid antibodies have been linked to increased miscarriage risk and reduced success rates in ART, particularly in intracytoplasmic sperm injection (ICSI) cycles. Although levothyroxine supplementation is widely used, its benefits in euthyroid women remain uncertain. Recent studies suggest that gut microbiota may modulate immune function and affect fertility outcomes among women with autoimmune thyroid conditions. This narrative review synthesizes findings from a broad literature base of over 40 peer-reviewed publications published between 2010 and 2025, with 30 of the most relevant and methodologically robust studies selected for detailed analysis. The review integrates clinical, endocrine, immunological, and microbiome-related perspectives. The evidence supports the need for personalized fertility management in women with Hashimoto’s thyroiditis and highlights directions for future research into immune and microbiota-targeted therapies. Full article

(This article belongs to the Special Issue Advances in Reproductive Endocrinology and Fertility: From Bench to Bedside)

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14 pages, 1728 KiB

Open AccessArticle

Auto Machine Learning and Convolutional Neural Network in Diabetes Mellitus Research—The Role of Histopathological Images in Designing and Exploring Experimental Models

by Iulian Tătaru, Simona Moldovanu, Oana-Maria Dragostin, Carmen Lidia Chiţescu, Alexandra-Simona Zamfir, Ionut Dragostin, Liliana Strat and Carmen Lăcrămioara Zamfir

Biomedicines 2025, 13(6), 1494; https://doi.org/10.3390/biomedicines13061494 - 18 Jun 2025

Histopathological images represent a valuable data source for pathologists, who can provide clinicians with essential landmarks for complex pathologies. The development of sophisticated computational models for histopathological images has received significant attention in recent years, but most of them rely on free datasets. [...] Read more.

Histopathological images represent a valuable data source for pathologists, who can provide clinicians with essential landmarks for complex pathologies. The development of sophisticated computational models for histopathological images has received significant attention in recent years, but most of them rely on free datasets. Materials and Methods: Motivated by this drawback, the authors created an original histopathological image dataset that resulted from an animal experimental model, acquiring images from normal female rats/rats with experimentally induced diabetes mellitus (DM)/rats who received an antidiabetic therapy with a synthetic compound (AD_SC). Images were acquired from vaginal, uterine, and ovarian samples from both MD and AD_DC specimens. The experiment received the approval of the Medical Ethics Committee of the “Gr. T. Popa” University of Medicine and Pharmacy, Iași, Romania (Approval No. 169/22.03.2022). The novelty of the study consists of the following aspects. The first is the use of a diabetes-induced animal model to evaluate the impact of an antidiabetic therapy with a synthetic compound in female rats, focusing on three distinct organs of the reproductive system (vagina, ovary, and uterus), to provide a more comprehensive understanding of how diabetes affects female reproductive health as a whole. The second comprises image classification with a custom-built convolutional neural network (CB-CNN), the extraction of textural features (contrast, entropy, energy, and homogeneity), and their classification with PyCaret Auto Machine Learning (AutoML). Results: Experimental findings indicate that uterine tissue, both for MD and AD_DC, can be diagnosed with an accuracy of 94.5% and 85.8%, respectively. The Linear Discriminant Analysis (LDA) classifier features indicate a high accuracy of 86.3% when supplied with features extracted from vaginal tissue. Conclusions: Our research underscores the efficacy of classifying with two AI algorithms, CNN and machine learning. Full article

(This article belongs to the Special Issue Artificial Intelligence Applications in Cancer and Other Diseases)

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11 pages, 954 KiB

Open AccessArticle

Serum NT-ProBNP/Chloride Ratio Predicts Adverse Cardiovascular Outcomes in Patients with Acute Heart Failure

by Victor José Leal-Alcántara, Eder González-Macedo, Ana Cristina Maldonado-May, Alberto Santiago-Hernández, Eder Jonathan Amaro-Palomo, Sarai Hernandez-Pastrana, Anna Elisa Adib-Gracia, Rodrigo Gopar-Nieto, Daniel Sierra-Lara Martínez, José Luis Briseño-De la Cruz, Héctor González-Pacheco, Alexandra Arias-Mendoza and Diego Araiza-Garaygordobil

Biomedicines 2025, 13(6), 1493; https://doi.org/10.3390/biomedicines13061493 - 18 Jun 2025

Background: Heart failure (HF) is a public health issue. It represents the second most common cause of hospitalization and the leading cause in individuals over 60 years old. Tools that predict adverse outcomes in patients with HF are needed. Objective: This study [...] Read more.

Background: Heart failure (HF) is a public health issue. It represents the second most common cause of hospitalization and the leading cause in individuals over 60 years old. Tools that predict adverse outcomes in patients with HF are needed. Objective: This study analyzed the prognostic role of the serum NT-proBNP/chloride ratio as a predictor of major cardiovascular events in patients with acute decompensated HF. Methods: Patients with a confirmed diagnosis of acute decompensated heart failure were retrospectively enrolled in the study; admission NT-proBNP/chloride ratio was used to stratify patients above or below the median (>/<83). The primary composite endpoint consisted of cardiovascular mortality, decompensated HF readmission, and unplanned emergency department visits. Results: A total of 197 individuals were included, of whom 100 (50.7%) were classified above and 97 (49.2%) below the median. Patients showing a high ratio had a lower LVEF (31 vs. 39%), a higher proportion of previous MI (30 vs. 15%), a lower diastolic blood pressure (73 vs. 80 mmHg), and higher BUN (38 vs. 23 mg/dL) and creatinine (1.6 vs. 1.1 mg/dL). After a follow-up period of 92 ± 3 days, 46 patients (23%) presented the primary endpoint; those with a high NT-proBNP/chloride ratio showed an increased risk (HR 3.18, 95% CI 1.55–6.52, p = 0.0015) of the primary endpoint. After multivariate analysis, only serum NT-proBNP/chloride ratio (p = 0.02) and diastolic pressure (0.037) remained significant. The area under the ROC curve for the NT-proBNP/chloride ratio for predicting the primary composite endpoint was significantly superior when compared with AUC for NT-proBNP or chloride alone. Conclusions: The serum NT-proBNP/chloride ratio is a novel, easy to use predictor of short- and medium-term cardiovascular events in patients with acute decompensated HF. Full article

(This article belongs to the Special Issue Heart Failure: New Diagnostic and Therapeutic Approaches)

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9 pages, 998 KiB

Open AccessArticle

Enteroviral Transverse Myelitis Presenting as Acute Ataxia in Children: A Case Series

by Luka Švitek, Dominik Ljubas, Nina Krajcar, Maja Vrdoljak Pažur, Ana Tripalo Batoš, Irena Tabain, Srđan Roglić and Lorna Stemberger Marić

Biomedicines 2025, 13(6), 1492; https://doi.org/10.3390/biomedicines13061492 - 18 Jun 2025

Background: Enteroviruses, members of the Picornaviridae family, typically cause asymptomatic or mild infections. However, they can also result in central nervous system (CNS) involvement, with transverse myelitis (TM) occurring only on rare occasions. TM is a syndrome characterized by acute or subacute [...] Read more.

Background: Enteroviruses, members of the Picornaviridae family, typically cause asymptomatic or mild infections. However, they can also result in central nervous system (CNS) involvement, with transverse myelitis (TM) occurring only on rare occasions. TM is a syndrome characterized by acute or subacute spinal cord dysfunction, leading to neurological deficits below the level of the lesion. Case report: We report a case series of eight pediatric patients admitted over a three-month period, June to August 2024. All patients presented with ataxia and/or other neurological symptoms, alongside abnormal cerebrospinal fluid (CSF) findings. Although ataxia is commonly associated with cerebellitis, magnetic resonance imaging (MRI) in this cohort revealed findings consistent with TM. Notably, all patients demonstrated similar MRI abnormalities. The onset of symptoms occurred over a short time during an enterovirus epidemic. Enteroviral RNA was detected, or the virus was isolated in seven patients, while one patient had a close epidemiological link to the virus. All patients achieved full recovery following immunomodulatory therapy. Conclusions: This case series underscores that ataxia may be an atypical symptom associated with TM. Furthermore, there was a notable distinction between the clinical presentation and neuroradiological findings. Immunomodulatory therapy with immunoglobulins and corticosteroids has been shown to be effective and safe, supporting the hypothesis of an immune-mediated pathogenesis in these patients. Full article

(This article belongs to the Special Issue Pathogenesis, Diagnosis and Treatment of Infectious Diseases)

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24 pages, 3748 KiB

Open AccessArticle

Prescribing Antidiabetic Medications Among GPs in Croatia—A Real-Life Cross-Sectional Study

by Tomislav Kurevija, Ema Schönberger, Matea Matić Ličanin, Ines Bilić-Ćurčić, Ljiljana Trtica-Majnarić and Silvija Canecki-Varžić

Biomedicines 2025, 13(6), 1491; https://doi.org/10.3390/biomedicines13061491 - 17 Jun 2025

Background: Advances in the treatment of type 2 diabetes (T2D) in recent decades have been primarily focused on its broader understanding in the context of the possibility of preventing the development and progression of the disease and of cardiovascular (CV) complications. Nevertheless, [...] Read more.

Background: Advances in the treatment of type 2 diabetes (T2D) in recent decades have been primarily focused on its broader understanding in the context of the possibility of preventing the development and progression of the disease and of cardiovascular (CV) complications. Nevertheless, worldwide research indicates that individuals with T2D are still under-regulated, both in terms of glycemic control and in preventing CV complications. The aim of this study was to examine Croatian general practitioners (GPs)’ practice and patterns in prescribing antidiabetic medications and their understanding of guidelines. Methods: Research was conducted using a self-designed anonymous survey, which was delivered to the e-mail addresses of GPs throughout Croatia in digital format. Respondents were solely GPs, without any restrictions with regard to their characteristics. Data on the number of individuals diagnosed with T2D and prescribed a specific medication were based on declarations by respondents from their e-health records. Results: Approximately 59% of individuals with T2D are cared for solely by GPs. In terms of achieving targeted values of HbA1c, 47% of individuals with T2D are well regulated. Almost all the respondents claim that they review prescribed T2D therapy at least once a year. A total of 47.6% of respondents have read and entirely understood the EASD/ADA guidelines, but 58.3% apply the dual principles of controlling HbA1c levels and CV risk in the treatment of T2D. In individuals with associated CV comorbidity, SGLT2ins were the most frequently prescribed. Conclusions: The results indicate that Croatian GPs are still inclined to apply outdated paradigms of T2D treatment but that they are gradually accepting new regimens of care and recommendations for prescribing novel, more effective medications. Full article

(This article belongs to the Special Issue Diabetes: Comorbidities, Therapeutics and Insights (2nd Edition))

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14 pages, 2539 KiB

Open AccessArticle

Sinusoidal Extremely Low-Frequency Electromagnetic Stimulation (ELF-EMS) Promotes Angiogenesis In Vitro

by Lena Perez Font, Amanda Moya-Gomez, Hannelore Kemps, Ivo Lambrichts, Jean-Michel Rigo, Bert Brône and Annelies Bronckaers

Biomedicines 2025, 13(6), 1490; https://doi.org/10.3390/biomedicines13061490 - 17 Jun 2025

Background/Objectives: Angiogenesis is the multistep process of the formation of new blood vessels. It is beneficial in scenarios that require tissue repair and regeneration, such as wound healing, bone fracture repair, and recovery from ischemic injuries like stroke, where new blood vessel [...] Read more.

Background/Objectives: Angiogenesis is the multistep process of the formation of new blood vessels. It is beneficial in scenarios that require tissue repair and regeneration, such as wound healing, bone fracture repair, and recovery from ischemic injuries like stroke, where new blood vessel formation restores oxygen and nutrient supply to damaged areas. Extremely low-frequency electromagnetic stimulation (ELF-EMS), which involves electromagnetic fields in the frequency range of 0–300 Hz, have been shown to reduce ischemic stroke volume by improving cerebral blood flow and recovery effects that are dependent on eNOS. Based on previous results, we herein explore the effects of ELF-EMS treatment (13.5 mT/10 and 60 Hz) on the activation of angiogenic processes in vitro in homeostatic conditions. Methods: Using human microvascular endothelial cells (HMEC-1), we studied cell proliferation, migration, and tube formation in vitro, as well as nitric oxide production and the effect of calcium and nitric oxide (NO) on these processes. Moreover, blood vessel formation was studied using a chicken chorioallantoic membrane (CAM) assay. Results: Our results showed that ELF-EMS increases proliferation, tube formation, and both the migration and transmigration of these cells, the latter of which was mediated via NO. In turn, calcium inhibition decreased ELF-EMF-induced NO production. Furthermore, ELF-EMS significantly increased blood vessel formation in the CAM assay. Conclusions: Our results indicated that ELF-EMS exposure (13.5 mT/10 and 60 Hz) significantly induces angiogenesis in vitro and in ovo, underscoring its potential application in the treatment of conditions characterized by insufficient blood supply. Full article

(This article belongs to the Section Cell Biology and Pathology)

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28 pages, 6060 KiB

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Identification Exploring the Mechanism and Clinical Validation of Mitochondrial Dynamics-Related Genes in Membranous Nephropathy Based on Mendelian Randomization Study and Bioinformatics Analysis

by Qiuyuan Shao, Nan Li, Huimin Qiu, Min Zhao, Chunming Jiang and Cheng Wan

Biomedicines 2025, 13(6), 1489; https://doi.org/10.3390/biomedicines13061489 - 17 Jun 2025

Background: Membranous nephropathy (MN), a prevalent glomerular disorder, remains poorly understood in terms of its association with mitochondrial dynamics (MD). This study investigated the mechanistic involvement of mitochondrial dynamics-related genes (MDGs) in the pathogenesis of MN. Methods: Comprehensive bioinformatics analyses—encompassing Mendelian randomization, machine-learning [...] Read more.

Background: Membranous nephropathy (MN), a prevalent glomerular disorder, remains poorly understood in terms of its association with mitochondrial dynamics (MD). This study investigated the mechanistic involvement of mitochondrial dynamics-related genes (MDGs) in the pathogenesis of MN. Methods: Comprehensive bioinformatics analyses—encompassing Mendelian randomization, machine-learning algorithms, and single-cell RNA sequencing (scRNA-seq)—were employed to interrogate transcriptomic datasets (GSE200828, GSE73953, and GSE241302). Core MDGs were further validated using reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Results: Four key MDGs—RTTN, MYO9A, USP40, and NFKBIZ—emerged as critical determinants, predominantly enriched in olfactory transduction pathways. A nomogram model exhibited exceptional diagnostic performance (area under the curve [AUC] = 1). Seventeen immune cell subsets, including regulatory T cells and activated dendritic cells, demonstrated significant differential infiltration in MN. Regulatory network analyses revealed ATF2 co-regulation mediated by RTTN and MYO9A, along with RTTN-driven modulation of ELOA-AS1 via hsa-mir-431-5p. scRNA-seq analysis identified mesenchymal–epithelial transitioning cells as key contributors, with pseudotime trajectory mapping indicating distinct temporal expression profiles: NFKBIZ (initial upregulation followed by decline), USP40 (gradual fluctuation), and RTTN (persistently low expression). RT-qPCR results corroborated a significant downregulation of all four genes in MN samples compared to controls (p < 0.05). Conclusions: These findings elucidate the molecular underpinnings of MDG-mediated mechanisms in MN, revealing novel diagnostic biomarkers and therapeutic targets. The data underscore the interplay between mitochondrial dynamics and immune dysregulation in MN progression, providing a foundation for precision medicine strategies. Full article

(This article belongs to the Section Gene and Cell Therapy)

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