Molecular Biology Reports最新文献

The incorporation of organoids with immune cells in co-culture systems signifies a groundbreaking advancement in the fields of cancer research and immunology. These three-dimensional models, derived from primary tumor specimens or stem cells, provide a more accurate representation of the tumor microenvironment (TME) than conventional two-dimensional cultures or animal models. This enhanced model allows for a thorough examination of the intricate interactions between cancer cells and the immune system. Although the success rates for organoid initiation can vary, averaging 36.8% across 13 different tumor types, successful organoid establishment enables the co-culture with a variety of immune cells, such as T cells, tumor-infiltrating lymphocytes (TILs), peripheral blood mononuclear cells (PBMCs), macrophages, dendritic cells, and natural killer (NK) cells. This platform enables the study of immune responses to cancer, mechanisms of immune evasion, and the influence of the TME on immune activation and suppression. The review emphasizes research involving intestinal, pancreatic, brain, liver, and cervical organoids, highlighting their role in elucidating disease mechanisms, assessing the effectiveness of immunotherapies (including checkpoint inhibitors and therapeutic vaccines), and conducting preclinical drug evaluations. Notable examples include modeling graft-versus-host disease with intestinal organoids, investigating the influence of DCLK1 on immunosuppression in pancreatic cancer, evaluating the effectiveness of engineered T cells against neuroblastoma using brain organoids, and analyzing the effects of cancer-associated fibroblasts on drug responses in colon cancer. Additionally, the potential of organoids in vaccine development and testing, particularly for influenza and other viral infections, is examined, demonstrating their utility in assessing immune responses and vaccine effectiveness. Despite existing challenges, such as the relatively low efficiency of organoid generation and the complexities involved in fully mimicking the TME, ongoing technological innovations, including tumor-on-chip systems and enhanced matrix materials, are expected to improve the functionality and clinical applicability of these advanced in vitro models.

Diabetic nephropathy (DN) is a primary contributor to end-stage renal disease, and non-coding RNAs (ncRNAs) extracted from urinary extracellular vesicles (uEVs) are being investigated as prospective biomarkers. This study reviewed the expression patterns, clinical associations, and diagnostic efficacy of them in DN cases. Following the PRISMA 2020 guidelines, PubMed, Embase, and Web of Science were systematically searched up to March 15, 2025 (PROSPERO: CRD420251039771). Eligible studies included primary human observational studies comparing uEV-derived ncRNAs in DN patients with non-DN diabetic subjects and healthy controls. Information on ncRNA expression, how it relates to clinical factors, and its performance as a diagnostic test was included. Bias assessment was done using the ROBINS-I tool. Twenty-four studies met the inclusion criteria. A total of 197 uEV-derived miRNAs were identified, while only one study evaluated other ncRNA species (four lncRNAs and two circRNAs). These miRNAs exhibited significant relationships with renal function markers, such as uACR, eGFR, serum creatinine, and BUN. Different miRNA signatures, such as miR-30a, miR-24-3p, and miR-27b-3p, were associated with stages of albuminuria. There were also associations between some miRNAs and glycemic indices, blood pressure, and lipid profiles. Individual miRNAs showed strong diagnostic efficacy, with miR-636, miR-34a, and miR-15b achieving AUCs of over 0.88. Combination biomarker panels significantly improved the ability to distinguish between DN stages. These results validate uEV-derived ncRNAs as sensitive indicators of DN development and prospective noninvasive biomarkers. However, standardized analytical procedures are necessary to make results more reproducible.

Purpose: Interleukin 1 receptor-associated kinase 1, 4 (IRAK 1/4) inhibitor exerts anti-inflammatory and immuno-modulatory effects; however, its role in high-fat diet-induced vascular dysfunction and cognitive impairment is not known, and therefore investigated in the present study.

Method & results: Animals were fed either a high-fat diet (60% Kcal fat) or a chow diet (10% Kcal fat) for 12 weeks to induce hyperlipidemia and weight gain. High-fat diet-fed animals were then treated with vehicle, IRAK1/4 inhibitor (2.2 mg/kg, i.p.) and a reference drug, Orlistat (20 mg/kg, oral gavage), for 4 additional weeks. Protein levels were assessed by ELISA or Western blotting, and mRNA by RT-PCR. IRAK1/4 inhibitor and reference drug, Orlistat treatment, prevented HFD-induced increase in body weight gain, fasting blood glucose and plasma lipids, improved discrimination between the familiar and the novel arm in the Y-Maze test, alleviated percent avoidance in two-way active avoidance, and freezing percent in contextual fear conditioning test. The treatments attenuated the levels of systemic inflammatory cytokines IL-1β, CRP, as well as TNF-α, IL-6 and protein expression of Iba-1, GFAP, HIF-1α, and restored the BDNF levels in the pre-frontal cortex of HFD-fed treated mice. IRAK 1/4 inhibitor exerted these effects by blocking proteasomal degradation of IκB-α protein in the pre-frontal cortex of HFD-treated mice. In addition, the treatments prevented HFD-induced increase in vascular ICAM-1, VCAM-1, MCP-1, COX-1 and COX-2 mRNA expression, and restored vascular eNOS mRNA levels as well as the Acetylcholine (300 ρM-300 μM) induced relaxations of PE (1 µM) pre-contracted aortic rings.

Conclusion: IRAK1/4 inhibitor attenuates HFD-induced inflammation, vascular dysfunction and cognitive impairment in obese mice.

Obesity-induced insulin resistance is an escalating global health challenge that substantially contributes to the development of metabolic disorders, including type 2 diabetes mellitus and cardiovascular disease. This narrative review critically examines the molecular and cellular mechanisms linking excess adiposity to impaired insulin action, with a particular focus on adipose tissue dysfunction, chronic low-grade inflammation, and oxidative stress. A comprehensive literature search was conducted using PubMed, ScienceDirect, and Google Scholar, covering preclinical and clinical studies published primarily over the past two decades. Evidence indicates that adipocyte hypertrophy and hypoxia promote excessive free fatty acid release, ectopic lipid accumulation, and lipotoxicity, thereby disrupting insulin signalling pathways. Numerous clinical studies report that obesity triggers chronic, low-grade inflammation in the liver and pancreas, activating pathways such as NF-κB and SOCS proteins, thereby disrupting insulin signalling. Concurrently, obesity-associated inflammation drives immune cell infiltration and macrophage polarization toward a pro-inflammatory phenotype, further exacerbating insulin resistance and metabolic dysregulation. This review also synthesizes current therapeutic strategies targeting these mechanisms, including insulin-sensitizing agents, anti-inflammatory therapies, glucagon-like peptide-1 receptor agonists, and sodium-glucose cotransporter 2 inhibitors, as well as emerging approaches. Future perspectives highlight the growing relevance of personalized medicine, pharmacogenomics, digital health tools, and gene-based interventions in improving therapeutic precision. A deeper understanding of these interconnected pathways is essential for developing effective strategies to mitigate obesity-related insulin resistance and its global metabolic consequences.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a significant public health concern due to its resistance to multiple antibiotics, primarily mediated by the gene mecA, which encodes penicillin-binding protein 2a (PBP2a), considering the growing global demand for novel antimicrobial strategies.

Materials and methods: A clinical MRSA strain harboring the gene mecA was identified. Antimicrobial susceptibility testing was performed to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Shirazi thyme (Zataria multiflora) essential oil. The strain's susceptibility to multiple antibiotics was assessed and the synergistic effect of Z. multiflora essential oil and cefoxitin was also evaluated. Gas chromatography-mass spectrometry (GC-MS) was employed to identify the bioactive compounds in the essential oil. Molecular docking studies were performed to evaluate the competitive binding affinity of those compounds to PBP2x.

Results: The MRSA strain exhibited resistance to all tested antibiotics except linezolid. The MIC and MBC values for Z. multiflora essential oil were 3.125 mg.mL^- 1 and 6.25 mg.mL^- 1, respectively, for the reference S. aureus strain (ATCC 25923), and 6.25 mg.mL^- 1 for both MIC and MBC for the clinical MRSA strain. The synergy assay demonstrated an enhanced inhibition zone for cefoxitin in combination with Z. multiflora essential oil, indicating a synergistic interaction. Molecular docking studies revealed strong binding interactions between spathulenol, isospathulenol, and aromadendrene comparable to clinically used β-lactam antibiotics.

Conclusion: The findings highlight the significant antibacterial activity of Z. multiflora essential oil against MRSA and its potential to enhance the efficacy of cefoxitin through synergistic interactions among the inhibitors of natural origin for PBP2x.

Background: Infertility affects 50-80 million individuals globally (WHO), with male factors, particularly abnormal semen parameters, contributing to 50% of cases. Among these, impaired sperm motility (asthenozoospermia, AZ) critically compromises fertilization potential. This study aimed to identify molecular biomarkers for AZ by investigating gene and competing endogenous RNA (ceRNA) networks associated with sperm motility.

Methods: The study population included 100 participants, including 50 AZ and 50 Normozoospermia (NZ, control), according to the semen criteria (WHO 2021). We integrated bioinformatics analyses, including differential gene expression profiling (Gene Expression Omnibus dataset), protein-protein interaction (PPI) networks, and miRNA-lncRNA-mRNA regulatory networks. Candidate biomarkers were experimentally validated via qRT-PCR and receiver operating characteristic (ROC) curve analyses in AZ patients.

Results: An integrated bioinformatics and experimental approach identified AKT1, hsa-miR-1286, and LINC00242 as a putative network associated with sperm motility that warrants further functional validation. QRT-PCR validation demonstrated significant downregulation of AKT1 and hsa-miR-1286, alongside upregulation of LINC00242 in AZ samples. Expression levels of AKT1 and hsa-miR-1286 showed positive correlations with sperm motility, whereas LINC00242 exhibited a strong negative correlation. ROC curve analysis further suggests preliminary diagnostic potential of these molecules.

Conclusions: Key findings suggested that AKT1, hsa-miR-1286, and linc00242 may represent preliminary diagnostic molecules, with the hsa-miR-1286/linc00242/AKT1 relationship emerging as a putative expression associated network linked to sperm motility.