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Keywords = differential gene expression analysis

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16 pages, 1951 KB  
Article
Effects of Stocking Density on Growth Performance, Physiological Responses, and Transcriptomic Profile of Penaeus monodon
by Jianzhi Shi, Wenzhe Li, Song Jiang, Hongshan Diao, Jianhua Huang, Lishi Yang, Yundong Li, Yangyang Ding, Yafei Duan, Xu Chen, Qibin Yang and Falin Zhou
Int. J. Mol. Sci. 2026, 27(3), 1505; https://doi.org/10.3390/ijms27031505 - 3 Feb 2026
Abstract
Stocking density plays an important role in P. monodon aquaculture. However, its underlying molecular mechanisms remain unknown. In this work, after a 40-day culture period, we studied the effects of different stocking densities (G1: 50, G2: 100, G3: 150 ind/m3) on [...] Read more.
Stocking density plays an important role in P. monodon aquaculture. However, its underlying molecular mechanisms remain unknown. In this work, after a 40-day culture period, we studied the effects of different stocking densities (G1: 50, G2: 100, G3: 150 ind/m3) on P. monodon, assessing their growth performances, physiological indexes, and transcriptome profiles. It was found that increased stocking density greatly decreased the growth performance and survival rate as well as the level of immune enzymes (alkaline phosphatase, acid phosphatase) and antioxidant component (total antioxidant capacity, superoxide dismutase, reduced glutathione) activities. Transcriptomic analysis showed that there are 2284 differentially expressed genes in all groups. Enrichment analysis of WGCNA results indicated that the green module associated with G2 was enriched for amino acid metabolism, lipid metabolism and immunity-related pathway. Furthermore, the G3 associated turquoise module was dominated by stress response, detoxification and energy metabolism pathways. Together, high stocking density causes the occurrence of oxidative stress, disturbance to immune system and alteration of metabolism profiles in P. monodon, whereas medium density (G2) is favorable to maintain physiological homeostasis. The results provide theoretical support to optimize aquaculture practice and contribute valuable information for subsequent studies. Full article
(This article belongs to the Special Issue Molecular Genetics and Genomics of Aquatic Crustaceans)
22 pages, 33722 KB  
Article
Integrated Transcriptomic and Histological Analysis of TP53/CTNNB1 Mutations and Microvascular Invasion in Hepatocellular Carcinoma
by Ignacio Garach, Nerea Hernandez, Luis J. Herrera, Francisco M. Ortuño and Ignacio Rojas
Genes 2026, 17(2), 190; https://doi.org/10.3390/genes17020190 - 3 Feb 2026
Abstract
Background/Objectives: Hepatocellular carcinoma (HCC) shows marked molecular and histopathological heterogeneity. Among the alterations most strongly associated with clinical outcome are mutations in TP53 and CTNNB1, as well as the presence of microvascular invasion (MVI). Although these factors are well established as [...] Read more.
Background/Objectives: Hepatocellular carcinoma (HCC) shows marked molecular and histopathological heterogeneity. Among the alterations most strongly associated with clinical outcome are mutations in TP53 and CTNNB1, as well as the presence of microvascular invasion (MVI). Although these factors are well established as prognostic indicators, how their molecular effects relate to tumor morphology remains unclear. In this work, we studied transcriptomic changes linked to TP53 and CTNNB1 mutational status and to MVI, and examined whether these changes are reflected in routine histology. Methods: RNA sequencing data from HCC samples annotated for mutations and vascular invasion were analyzed using differential expression analysis combined with machine learning-based feature selection to characterize the underlying transcriptional programs. In parallel, we trained a weakly supervised multitask deep learning model on hematoxylin and eosin-stained whole-slide images using slide-level labels only, without spatial annotations, to assess whether these features could be inferred from global histological patterns. Results: Distinct gene expression profiles were observed for TP53-mutated, CTNNB1-mutated, and MVI-positive tumors, involving pathways related to proliferation, metabolism, and invasion. Image-based models were able to capture morphological patterns associated with these states, achieving above-random discrimination with variable performance across tasks. Conclusions: Taken together, these results support the existence of coherent biological programs underlying key risk determinants in HCC and indicate that their phenotypic effects are, at least in part, detectable in routine histopathology. This provides a rationale for integrative morpho-molecular approaches to risk assessment in HCC. Full article
(This article belongs to the Special Issue AI and Machine Learning in Cancer Genomics)
21 pages, 1994 KB  
Article
Vitamin D Reprograms Non-Coding RNA Networksto Block Zika Virus in Human Macrophages
by Julieta M Ramírez-Mejía, Geysson Javier Fernandez and Silvio Urcuqui-Inchima
Pathophysiology 2026, 33(1), 15; https://doi.org/10.3390/pathophysiology33010015 - 3 Feb 2026
Abstract
Background: Zika virus (ZIKV), a mosquito-borne flavivirus, is associated with congenital malformations and neuroinflammatory disorders, highlighting the need to identify host factors that shape infection outcomes. Macrophages, key targets and reservoirs of ZIKV, orchestrate both antiviral and inflammatory responses. Methods: Vitamin D (VitD) [...] Read more.
Background: Zika virus (ZIKV), a mosquito-borne flavivirus, is associated with congenital malformations and neuroinflammatory disorders, highlighting the need to identify host factors that shape infection outcomes. Macrophages, key targets and reservoirs of ZIKV, orchestrate both antiviral and inflammatory responses. Methods: Vitamin D (VitD) has emerged as a potent immunomodulator that enhances macrophage antimicrobial activity and regulates inflammation. To investigate how VitD shapes macrophage responses to ZIKV, we reanalyzed publicly available RNA-seq and miRNA-seq datasets from monocyte-derived macrophages (MDMs) of four donors, differentiated with or without VitD and subsequently infected with ZIKV. Results: Differential expression analysis identified long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs integrated into competing endogenous RNA (ceRNA) networks. In VitD-conditioned and ZIKV-infected MDMs, 65 lncRNAs and 23 miRNAs were significantly modulated. Notably, lncRNAs such as HSD11B1-AS1, Lnc-FOSL2, SPIRE-AS1, and PCAT7 were predicted to regulate immune and metabolic genes, including G0S2, FOSL2, PRELID3A, and FBP1. Among the miRNAs, let-7a and miR-494 were downregulated, while miR-146a, miR-708, and miR-378 were upregulated, all of which have been previously implicated in antiviral immunity. Functional enrichment analysis revealed pathways linked to metabolism, stress responses, and cell migration. ceRNA network analysis suggested that SOX2-OT and SLC9A3-AS1 may act as molecular sponges, modulating regulatory axes relevant to immune control and viral response. Conclusions: Despite limitations in sample size and experimental validation, this study provides an exploratory map of ncRNA–mRNA networks shaped by VitD during ZIKV infection, highlighting candidate molecules and pathways for further studies on host–virus interactions and VitD-mediated immune regulation. Full article
(This article belongs to the Section Cellular and Molecular Mechanisms)
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25 pages, 6314 KB  
Article
BCL2A1high CD8+ T Cells Are a Survival-Associated Predictor of Immune Checkpoint Blockade Response in Lung Adenocarcinoma
by Hoang Minh Quan Pham, Po-Hao Feng, Chia-Ling Chen, Kang-Yun Lee and Chiou-Feng Lin
Diagnostics 2026, 16(3), 475; https://doi.org/10.3390/diagnostics16030475 - 3 Feb 2026
Abstract
Background: Immune checkpoint blockade (ICB) has revolutionized lung adenocarcinoma (LUAD) therapy, yet predictive bio-markers remain suboptimal. We hypothesized that BCL2A1 expression in CD8+ T cells may reflect immune endurance and complement PD-L1 in predicting ICB response. Methods: Integrating bulk and [...] Read more.
Background: Immune checkpoint blockade (ICB) has revolutionized lung adenocarcinoma (LUAD) therapy, yet predictive bio-markers remain suboptimal. We hypothesized that BCL2A1 expression in CD8+ T cells may reflect immune endurance and complement PD-L1 in predicting ICB response. Methods: Integrating bulk and single-cell RNA-seq across multiple LUAD cohorts, this study performed differential expression, survival, and pathway analyses in a discovery cohort (n = 60) and validated findings across five independent cohorts (n = 126). Results: Single-cell profiling identified BCL2A1 enrichment in tissue-resident memory and proliferating subsets that appeared preferentially expanded in responders; cell–cell communication analysis revealed that BCL2A1high CD8+ T cells exhibited significantly enhanced outgoing signaling capacity (p = 0.0278), with proliferating subsets serving as intra-CD8+ coordination hubs and MIF pathway interactions achieving the highest intensity among all axes examined. BCL2A1 was significantly upregulated in responders (FDR < 0.05) and associated with improved ICB survival (HR = 0.43, p < 0.05), but not in non-ICB settings, suggesting treatment-specific prognostic relevance. A tri-marker model integrating BCL2A1, PD-L1 (CD274), and a 27-gene HOT score demonstrated favorable predictive performance (AUC = 0.826 discovery; macro-AUC = 0.774 validation), outperforming PD-L1 alone (AUC = 0.706) and established signatures including TIDE, IPS, TIS, and IFNG. Cross-platform simulations suggested high reproducibility (ρ = 0.982–0.993). Conclusions: These findings suggest BCL2A1 may serve as a bio-marker of CD8+ T-cell survival and enhanced intercellular coordination, and its integration with PD-L1 and immune activation markers may yield a reproducible ICB response predictor, pending clinical validation. Full article
(This article belongs to the Section Pathology and Molecular Diagnostics)
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17 pages, 4912 KB  
Article
Transcriptomic Analysis of Gene Expression Patterns in the Cecal Tissue of Liangshan Yanying Chickens and Arbor Acres (AA) Chickens Before 28 Days of Age
by Zengwen Huang, Jing Wang, Chaoyun Yang and Runjin Wang
Animals 2026, 16(3), 474; https://doi.org/10.3390/ani16030474 - 3 Feb 2026
Abstract
To dissect the molecular mechanisms underlying chicken cecal development, this study used Liangshan Yanying chickens (a local slow-growing breed) and Arbor Acres (AA) chickens (a fast-growing breed) as experimental models. Cecal tissues were collected from healthy chickens at 1, 14, and 28 days [...] Read more.
To dissect the molecular mechanisms underlying chicken cecal development, this study used Liangshan Yanying chickens (a local slow-growing breed) and Arbor Acres (AA) chickens (a fast-growing breed) as experimental models. Cecal tissues were collected from healthy chickens at 1, 14, and 28 days of age (n = 10 per breed per day of age) to measure cecal length and perform transcriptome sequencing. Through the screening of differentially expressed genes (DEGs), functional enrichment analysis, construction of protein–protein interaction (PPI) networks, and qRT-PCR validation, temporal changes in cecal development between the two breeds were systematically compared. Results showed that cecal length of both breeds increased significantly with age (p < 0.05), with significant differences between breeds. A total of 18 high-quality samples were obtained from transcriptome analysis (Q30 ≥ 93%), with a mapping efficiency of 86.2–90.5%. The number of DEGs was highest between 1 and 28 days of age (1844 DEGs in Liangshan Yanying chickens and 1747 DEGs in AA chickens), and the number of inter-breed DEGs reached 2133 at 28 days of age. A total of 70 DEGs with consistent expression trends were identified (22 upregulated and 48 downregulated), which were enriched in Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways such as “B cell activation”, “peptide transport”, and “bile acid metabolism”. qRT-PCR validation indicated that the expression trends of genes (e.g., CD79B, IRF4) were highly consistent with sequencing results (R2 = 0.91). PPI network analysis suggested that SLC15A1, ACE, and ENPEP were key hub genes, forming a “transport–metabolism” synergistic module. This study reveals the temporal dynamics of chicken cecal development and the molecular basis of inter-breed differences, providing a theoretical foundation for broiler genetic improvement. Full article
(This article belongs to the Section Poultry)
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16 pages, 2627 KB  
Article
Single-Cell Mapping Reveals MIF-Centered Immunoregulatory Networks in Colorectal Cancer
by Marios Gkoris, Ilias Georgakopoulos-Soares and Apostolos Zaravinos
Int. J. Mol. Sci. 2026, 27(3), 1496; https://doi.org/10.3390/ijms27031496 - 3 Feb 2026
Abstract
Colorectal cancer (CRC) progression is strongly shaped by the tumor microenvironment (TME), where complex interactions between epithelial, immune, and stromal cells orchestrate immune suppression and tumor evolution. To dissect these relationships at single-cell resolution, we analyzed CRC scRNA-seq datasets using Seurat for data [...] Read more.
Colorectal cancer (CRC) progression is strongly shaped by the tumor microenvironment (TME), where complex interactions between epithelial, immune, and stromal cells orchestrate immune suppression and tumor evolution. To dissect these relationships at single-cell resolution, we analyzed CRC scRNA-seq datasets using Seurat for data integration and CellChat for ligand–receptor inference. We identified extensive cellular heterogeneity within the TME, dominated by CMS2/CMS3 epithelial states, SPP1+ tumor-associated macrophages, diverse T-cell subsets, and CXCR4+ B cells. Communication analysis revealed MIF-centered signaling—including MIF–CD74–CXCR4 and MIF–CD74–CD44—as the predominant axis linking tumor epithelial cells with T cells, B cells, and macrophage subpopulations. CMS3 epithelial cells displayed particularly strong connectivity to SPP1+ macrophages and cytotoxic lymphocytes through both MIF- and APP–CD74-mediated pathways. Differential gene expression confirmed elevated levels of MIF, CD74, CD44, and SPP1 in tumor tissues, while pathway enrichment analyses highlighted cytokine signaling, antigen presentation, and chemokine-regulated immune modulation as key biological processes. Collectively, our study provides a high-resolution map of CRC intercellular communication and identifies MIF-CD74-associated signaling as a central immunoregulatory hub with potential relevance for therapeutic targeting and biomarker development. Full article
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29 pages, 1572 KB  
Article
Bioinformatic Analysis of Contrasting Expression Patterns and Molecular Interactions of TIMPs in Breast Cancer: Implications for Tumor Progression and Survival
by Lorena Cayetano-Salazar, Jhactcidi Jackeline García-López, Dania A. Nava-Tapia, Eymard Hernández-López, Caroline Weinstein-Oppenheimer, Julio Ortiz-Ortiz, Marco Antonio Leyva-Vázquez, Miguel Ángel Mendoza-Catalán, Adán Arizmendi-Izazaga and Napoleón Navarro-Tito
Pathophysiology 2026, 33(1), 13; https://doi.org/10.3390/pathophysiology33010013 - 2 Feb 2026
Abstract
Background/Objectives: Although tissue inhibitors of metalloproteinases (TIMPs) are key regulators in breast cancer, their differential expression, clinical relevance, and molecular roles remain unclear. This study aimed to compare the expression patterns of the four TIMPs in breast cancer and evaluate their molecular interactions [...] Read more.
Background/Objectives: Although tissue inhibitors of metalloproteinases (TIMPs) are key regulators in breast cancer, their differential expression, clinical relevance, and molecular roles remain unclear. This study aimed to compare the expression patterns of the four TIMPs in breast cancer and evaluate their molecular interactions and associated pathways through an integrated bioinformatic analysis. Methods: The expression of TIMPs and their correlations with MMPs were analyzed using the TCGA PanCancer, cBioPortal, and GEO datasets. Associations between TIMP expression and overall survival were assessed in the TCGA Breast Invasive Carcinoma PanCancer cohort. Pathway enrichment analysis was performed using GO, KEGG, and DAVID. The relationships between immune cell infiltration, stromal cells, and TIMP expression were assessed using the EPIC algorithm. Statistical analyses were performed using R. Results: TIMP1 was the only inhibitor overexpressed in breast tumors and showed significant associations with the Luminal B, HER2, TNBC, and normal-like subtypes, along with a modest increase across stages. TIMP2, TIMP3, and TIMP4 were downregulated in tumors. High expression of TIMP1 and TIMP4 correlated with better overall survival. TIMP1-associated genes were enriched in NF-kappa and PI3K–Akt signaling and actin cytoskeleton components. TIMP2 was linked to Hedgehog and MAPK pathways and actin-related elements. TIMP3 correlated with Hedgehog and PI3K–Akt signaling, DNA damage response, and membrane components. TIMP4 was associated with VEGF, MAPK, PI3K–Akt, DNA damage pathways, and actin organization. TIMP2 showed strong positive correlations with MMP2 and MMP14, while TIMP4 showed negative correlations with MMP1 and MMP9. Interestingly, we found a strong positive correlation between TIMP2 and TIMP3 with ADAM12, as well as between TIMP2 and TIMP3 with ADAM10, and negative correlations with ADAM15. The differential expression of TIMPs favors greater infiltration of immune cells related to tumor progression and poor prognosis in breast cancer patients. Conclusions: TIMPs display contrasting expression profiles and distinct pathway associations in breast cancer. TIMP1 emerges as the only consistently overexpressed inhibitor, while TIMP4 appears as a promising prognostic marker with unique MMP correlations that may influence tumor behaviors. Full article
(This article belongs to the Section Cellular and Molecular Mechanisms)
17 pages, 1718 KB  
Article
Integrated Transcriptomic and Metabolomic Analysis Reveals Molecular Signatures Associated with Natural Degeneration of Puccinia striiformis f. sp. tritici
by Congying Yuan, Tianyu Long, Jiani Dong, Bingyu Yan, Tingxuan Chen, Yubin Zhang, Yuanhan Yan, Mengyu Cheng and Sitong Xue
Curr. Issues Mol. Biol. 2026, 48(2), 169; https://doi.org/10.3390/cimb48020169 - 2 Feb 2026
Abstract
Stripe rust of wheat, caused by the obligate biotrophic fungus Puccinia striiformis f. sp. tritici (Pst), is a devastating disease. The natural degeneration and viability loss of Pst urediniospores directly impact its dispersal and epidemic potential, yet the underlying molecular mechanisms remain unclear. [...] Read more.
Stripe rust of wheat, caused by the obligate biotrophic fungus Puccinia striiformis f. sp. tritici (Pst), is a devastating disease. The natural degeneration and viability loss of Pst urediniospores directly impact its dispersal and epidemic potential, yet the underlying molecular mechanisms remain unclear. This study aimed to systematically decipher the key molecular changes during the natural degeneration of Pst urediniospores using a multi-omics approach. We performed integrated transcriptomic (RNA-seq) and metabolomic (LC-MS) analyses on relatively purified fresh urediniospores (CC group) and those undergoing room-temperature-induced degeneration (CM group) of the prevalent Pst race CYR34. A total of 1622 differentially expressed genes (DEGs) and 382 differentially accumulated metabolites (DAMs) were identified. Transcriptomic analysis revealed significant downregulation of core energy and biosynthetic pathways, including ribosome biogenesis and oxidative phosphorylation. Metabolomic profiling showed that lipids and lipid-like molecules, along with organic acids and derivatives, constituted the major classes of altered metabolites. DAMs were primarily enriched in pathways such as “Metabolic pathways” and “ABC transporters.” Integrated analysis indicated a prevalent negative correlation pattern between gene expression levels and metabolite abundance. This study provides a systematic molecular landscape associated with Pst urediniospore degeneration, revealing characteristics concomitant with the suppression of energy metabolism and translation functions, thereby offering novel insights and a data foundation for understanding the mechanisms of viability maintenance and loss. Full article
(This article belongs to the Section Molecular Microbiology)
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30 pages, 6406 KB  
Article
Cell Line–Dependent Cell Death Pathways Induced by Thymoquinone in Colorectal Cancer Cells
by Natalia Kurowska, Maria Książek, Paulina Borkowska and Barbara Strzałka-Mrozik
Molecules 2026, 31(3), 512; https://doi.org/10.3390/molecules31030512 - 2 Feb 2026
Abstract
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, with resistance to 5-fluorouracil (5-FU) representing a major therapeutic challenge. Thymoquinone (TQ), a bioactive constituent of Nigella sativa, exhibits anticancer activity; however, the mechanisms underlying TQ-induced cell death appear to be highly [...] Read more.
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, with resistance to 5-fluorouracil (5-FU) representing a major therapeutic challenge. Thymoquinone (TQ), a bioactive constituent of Nigella sativa, exhibits anticancer activity; however, the mechanisms underlying TQ-induced cell death appear to be highly context dependent. This study aimed to characterize cell line-specific death pathways triggered by TQ in colorectal cancer models with distinct molecular backgrounds and differential responsiveness to 5-FU. Human CRC cell lines RKO (5-FU-sensitive) and SW1116 (poorly responsive), along with normal colon epithelial cells (CCD-841CoN), were treated with TQ, 5-FU, or their combination for 24 h. Cell viability, DNA fragmentation, caspase-3/7, -8, and -9 activity, cell death phenotypes, and expression of apoptosis- and necroptosis-related genes were evaluated using MTT assays, ELISA, luminescent assays, flow cytometry, and RT-qPCR. TQ significantly reduced viability in both CRC cell lines while exerting minimal cytotoxicity toward normal cells. In RKO cells, characterized by microsatellite instability (MSI), TQ induced DNA fragmentation, caspase activation, and transcriptional upregulation of pro-apoptotic genes, consistent with apoptosis-associated signaling. In contrast, SW1116 cells, which exhibit chromosomal instability (CIN) and reduced responsiveness to 5-FU, displayed decreased viability accompanied by suppressed caspase activity and predominant features of caspase-independent necrotic cell death. This differential response may be attributed to the CIN phenotype, which has been associated with impaired apoptotic signaling and enhanced tolerance to cytotoxic stress. Combined TQ and 5-FU treatment did not produce synergistic cytotoxicity, as confirmed by Bliss independence analysis, but revealed distinct, cell line-dependent death programs. These findings demonstrate that TQ modulates cell death execution in a molecular context-dependent manner rather than enhancing 5-FU efficacy through pharmacological synergy. Full article
(This article belongs to the Special Issue Natural Compounds in Modern Therapies, 3rd Edition)
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20 pages, 3317 KB  
Article
Study on the Effect of N-Carbamylglutamate (NCG) on Reproductive Performance and Regulation Mechanism of Primary Lake Sheep
by Tianli Gao, Chunyang Li, Juanshan Zheng, Yingpai Zhaxi, Yuan Cai, Rongxin Zang, Huixia Liu, Yanmei Yang, Sai Li, Xiaodi Shi and Chen Huang
Animals 2026, 16(3), 464; https://doi.org/10.3390/ani16030464 - 2 Feb 2026
Abstract
The aim of this study was to investigate the effects of dietary supplementation with 0.11% N-carbamylglutamate (NCG) during early pregnancy (0–90 days) on reproductive performance and fetal development, and to elucidate the underlying placental regulatory mechanisms in primiparous Hu sheep. Twenty-two 10-month-old sexually [...] Read more.
The aim of this study was to investigate the effects of dietary supplementation with 0.11% N-carbamylglutamate (NCG) during early pregnancy (0–90 days) on reproductive performance and fetal development, and to elucidate the underlying placental regulatory mechanisms in primiparous Hu sheep. Twenty-two 10-month-old sexually mature primiparous Hu sheep meeting the mating criteria were randomly assigned to two groups. The control group was fed a basal diet, while the NCG group received the basal diet supplemented with 0.11% NCG, with both feeding regimens maintained for 90 days. By measuring uterine and fetal growth indices, maternal plasma biochemical parameters, and amino acid levels, as well as assessing cotyledon indices and observing cotyledon morphology and histological structure, basic data related to placental function and fetal growth in pregnant ewes was collected. Combined with transcriptomic sequencing of maternal placental tissue, the mechanism by which NCG influences placental function and fetal growth and development in pregnant ewes was further investigated. The supplementation of NCG could increase the number of fetuses, total weight of fetuses, the number of corpus luteum and the ratio of fetuses to corpus luteum, but the difference was not significant (p > 0.05). The levels of plasma NO, inducible Nitric Oxide Synthase (iNOS) and several amino acids were significantly increased (p < 0.05). In ewes’ uteri, the average uterine weight, number of uterine glands, total cotyledon weight, and average weight per cotyledon were significantly increased (p < 0.05), whereas uterine mucosal thickness was markedly decreased. The Quantitative Real-time PCR (q-PCR) results for differentially expressed genes were consistent with those of transcriptomic analysis, showing significant changes in the expression levels of certain differentially expressed genes in maternal placental tissues. These changes regulated pathways such as vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF), phosphatidylinositol 3-kinase–protein kinase B (PI3K–AKT) signaling pathways and Mitogen-Activated Protein Kinase (MAPK) pathway, which are involved in angiogenesis, energy supply and metabolism, and somatic growth and development. Dietary supplementation with NCG during early pregnancy can significantly improve the reproductive performance of primiparous Hu sheep, optimize the intrauterine environment and nutrient supply, and thereby facilitate pregnancy maintenance and fetal development. The underlying mechanism may involve promoting endogenous arginine synthesis in ewes, increasing plasma levels of NO, arginine, and certain amino acids, which collectively validate the positive effects of NCG on the reproductive performance and growth of Hu sheep during early pregnancy at the molecular level. Full article
(This article belongs to the Section Animal Reproduction)
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24 pages, 13256 KB  
Article
Malva sylvestris Flower Extract Exhibits Antineoplastic Potential Against Human Colon Cancer Cell Lines and Induces CDK2 Transcript Instability via Plant miR160-5p
by Valentina Villani and Angelo Gismondi
Nutrients 2026, 18(3), 495; https://doi.org/10.3390/nu18030495 - 2 Feb 2026
Abstract
Background: Malva sylvestris (the common mallow) is an herbaceous species widely used in ethnobotanical practices to treat gastrointestinal, hepatic and urinary inflammation. Objectives: Despite these beneficial effects on human health, the antineoplastic potential of this plant has not yet been fully explored. [...] Read more.
Background: Malva sylvestris (the common mallow) is an herbaceous species widely used in ethnobotanical practices to treat gastrointestinal, hepatic and urinary inflammation. Objectives: Despite these beneficial effects on human health, the antineoplastic potential of this plant has not yet been fully explored. Thus, in the present study, two human colon cancer cell lines (i.e., HCT-116 and Caco-2) were treated with an extract obtained from M. sylvestris flowers (MFE), whose composition in terms of phytochemicals and microRNAs has been recently published by our research group, to explore its potential bioactivity. Methods/Results: MTT and Trypan blue assays demonstrated that MFE reduced tumour cell growth without causing significant cytotoxicity or apoptosis. Following the diphenylboric acid 2-aminoethyl ester-induced fluorescence of some plant metabolites, microscopy analysis proved that MFE components crossed the cell membranes, accumulating into nuclei. Wound assay and transwell tests documented that MFE was also able to reduce cell motility and invasiveness. In both cell lines qPCR experiments demonstrated that MFE caused the over-expression of factors, like VIMENTIN and E-CADHERIN, which negatively influence epithelial–mesenchymal transition in colon cancers. However, the effects of MFE appeared to be time-, dose- and cell type-dependent. In fact, the treatment induced senescence in P53-null Caco-2 cells (i.e., ROS, β-galactosidase and P21WAF1/Cip1) and a premise of differentiation (i.e., P27Kip1) in P53-wild-type HCT-116 cells, also via the CDK2/c-MYC/AKT axis, justifying its antiproliferative property. In parallel, the transfection of tumour cells with pure synthetic miR160b-5p—a microRNA identified in M. sylvestris flowers and predicted to target the human CDK2 transcript—resulted in gene silencing, thereby suggesting its central role in mediating the cross-kingdom effects of MFE on the investigated cancer models. Conclusions: Overall, these findings open new perspectives on the common mallow as a source of potential antimetastatic compounds and on the possible use of its plant microRNAs in the development of gene therapies. Full article
(This article belongs to the Special Issue Natural Active Substances and Cancer)
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26 pages, 13963 KB  
Article
Deciphering Drought Response Mechanisms in Oat Through Comprehensive Transcriptomic and Physiological Analysis
by Baiji Wang, Hang Yin, Xinyi Zhang, Xiangpeng Kong, Wenjie Zhao, Rui Qiu, Muzhapaer Tuluhong, Guowen Cui and Bing Li
Plants 2026, 15(3), 453; https://doi.org/10.3390/plants15030453 - 1 Feb 2026
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Abstract
Oat, an important cereal and forage crop, is significantly affected by drought stress during production. However, the molecular mechanisms underlying oat’s response to drought stress remain largely unknown. In this study, K-means clustering classified 28 oat varieties into drought-tolerant (Muda, Mengshi No. 1) [...] Read more.
Oat, an important cereal and forage crop, is significantly affected by drought stress during production. However, the molecular mechanisms underlying oat’s response to drought stress remain largely unknown. In this study, K-means clustering classified 28 oat varieties into drought-tolerant (Muda, Mengshi No. 1) and drought-sensitive (Heike, Haywire) groups, with grey relational analysis further verifying MS as the most drought-tolerant and HK as the most drought-sensitive variety. Under drought stress, drought-tolerant and drought-sensitive varieties showed notable differences in leaf chlorophyll content, osmoregulation substances, and the activities of antioxidant enzymes. Transcriptomic analysis showed that 1915 differentially expressed genes (DEGs) were shared among all comparisons between treatment groups and the control group. KEGG pathway analysis revealed enrichment in pathways such as plant–pathogen interactions, plant hormone signal transduction, and starch and sucrose metabolism. In the signal transduction of plant hormones, eight PP2C genes associated with ABA signaling were increased, indicating that oats might respond to drought by enhancing metabolic activities via the ABA signaling pathway. WGCNA identified gene modules significantly associated with physiological traits. Notably, Mantel tests revealed that six core genes exhibited a positive correlation with CAT activity in the drought-tolerant variety, while showing an opposite trend in the sensitive variety. This study provides insights into the mechanisms of drought tolerance in oats and aids in the molecular breeding of drought-tolerant varieties. Full article
(This article belongs to the Section Crop Physiology and Crop Production)
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21 pages, 4522 KB  
Article
Transcriptomic Exploration of Muscle Development and Fat Deposition Trait Diversity in Selected Indian Sheep Breeds: Implications for Meat Quality and Yield
by Navya Pothireddy, Mangalathu Rajan Vishnuraj, Kappala Vijaya Rachel, Peddapuram Baswa Reddy, Prashantha Chowdadenahalli Nagaraja, Ajay Ganesan, Shiva Shankar Kanneboyina, Krishnachaithanya Indiradevi and Sukhadeo Baliram Barbuddhe
Animals 2026, 16(3), 452; https://doi.org/10.3390/ani16030452 - 1 Feb 2026
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Abstract
Sheep’s meat production and quality are influenced by genetic and physiological factors that affect muscle development, growth, and fat deposition metabolism. However, the breed-specific transcriptional landscapes driving these traits in Indian sheep breeds, especially in Nellore (meat-type) and Deccani (wool-meat type) breeds are [...] Read more.
Sheep’s meat production and quality are influenced by genetic and physiological factors that affect muscle development, growth, and fat deposition metabolism. However, the breed-specific transcriptional landscapes driving these traits in Indian sheep breeds, especially in Nellore (meat-type) and Deccani (wool-meat type) breeds are remain unexplored. Therefore, this study aimed to investigate the differences in muscle growth and fat deposition between Nellore and Deccani breeds by integrating transcriptomic profiling, carcass characteristics, and histological analysis of longissimus dorsi muscle and liver tissues. Carcass assessment revealed higher Hot Carcass Weight (HCW), Cold Carcass Weight (CCW), Hot Carcass Yield (HCY) and Cold Carcass Yield (CCY), and larger myofibrillar cross-sectional area (p < 0.05), indicating enhanced musculature, which was observed in Nellore. Deccani showed elevated Intramuscular Fat (IMF) deposition (p < 0.05), indicating improved meat flavour/juiciness. Transcriptomic profiling revealed several Differentially Expressed Genes (DEGs) associated with meat quality and quantity traits. In Nellore, the genes WFIKKN2, FGFRL1, FKBP4, and IRF1 were upregulated, while the gene TAS1R2 was downregulated, leading to enhanced muscle development, superior carcass traits, thermotolerance, and immunity. While Deccani showed higher expression of lipid metabolism genes PLA2G4F, ACSL1, ACOX1, CPT1A, and PLIN1, which are linked to higher IMF content. Functional enrichment analysis revealed 46 significantly enriched GO terms for the DEGs (p < 0.05), including oxidoreductase activity, muscle development, etc. These outcomes demonstrate novel genetic markers and key biological insights into the regulation of muscle development, thermotolerance, immunity, and IMF for future validation in Indian sheep breeds. Full article
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16 pages, 1898 KB  
Article
Disorganization of Transcriptional Regulation and Alteration of Keratin Family Gene Expression in Hairy Ear Mice
by Byeongyong Ahn, Hojun Choi, Joori Yum, Dayoung Kim, Harris Lewin and Chankyu Park
Genes 2026, 17(2), 182; https://doi.org/10.3390/genes17020182 - 31 Jan 2026
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Abstract
Background: The hairy ear (Eh) mutation in heterozygous mice (Eh/+) results in elongated and additional ear hairs, along with altered pinna morphology compared to wild-type (+/+) mice. Previous studies suggest that disruption of the Hoxc gene cluster caused by [...] Read more.
Background: The hairy ear (Eh) mutation in heterozygous mice (Eh/+) results in elongated and additional ear hairs, along with altered pinna morphology compared to wild-type (+/+) mice. Previous studies suggest that disruption of the Hoxc gene cluster caused by the Eh inversion influences the hair growth cycle. Methods: To elucidate the molecular basis of this phenotype, we performed RNA-seq analysis on ear tissues from four-week-old Eh/+ and +/+ mice and compared their transcriptomic profiles. Results: Differential expression analysis identified 2092 genes, and subsequent Gene Ontology (GO) and overrepresentation analysis revealed significant alterations in hair growth-related processes, including the hair cycle and canonical keratinization in Eh/+ ears. Notably, numerous hair keratin and keratin-associated protein (Krtap) genes were markedly upregulated in Eh/+ mice. Validation by quantitative real-time PCR confirmed increased expression of randomly selected keratin genes (Krt34, Krt39, Krt71, Krt81, Krt84) and keratin-associated proteins (Krtap4-16 and Krtap22-2). In contrast, epithelial keratin genes such as Krt2 and Krt14 were downregulated in Eh/+ ears. In addition, genes associated with hair follicle growth, Car6 and Gprc5d, showed elevated expression, while Dab2, a telogen–anagen transition marker linked to hair follicle stem cell activation, was slightly increased at the telogen stage in Eh/+ compared with +/+ mice. Conclusions: These findings provide new insights into the role of Hoxc cluster genes in orchestrating the expression of hair keratin and Krtap genes and highlight potential regulatory mechanisms underlying the hairy ear phenotype. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
17 pages, 4832 KB  
Article
Identification of Key Genes Regulating Body Weight in Qingyuan Partridge Chickens During Development using RNA-Sequence Analysis
by Junyi Zhuang, Weifang Yang, Yanji Chen, Shuang Liu, Xucheng He, Jiguang Deng, Yucheng Zhang, Maiqing Zheng, Guiping Zhao, Jie Wen and Huanxian Cui
Genes 2026, 17(2), 179; https://doi.org/10.3390/genes17020179 - 31 Jan 2026
Viewed by 62
Abstract
Background: The Qingyuan partridge chicken is a high-quality local chicken breed in China. Its weight gain directly affects breeding efficiency. This study used RNA sequencing to analyze gene expression dynamics in the breast muscle tissue of Qingyuan partridge chickens at 1, 35, [...] Read more.
Background: The Qingyuan partridge chicken is a high-quality local chicken breed in China. Its weight gain directly affects breeding efficiency. This study used RNA sequencing to analyze gene expression dynamics in the breast muscle tissue of Qingyuan partridge chickens at 1, 35, 70, and 105 days of age. Methods: This study employed RNA-sequencing, integrated with differential expression analysis, weighted gene co-expression network analysis (WGCNA), and short time-series expression miner (STEM) analysis, to systematically investigate the transcriptomic dynamics in breast muscle tissue across four developmental stages. Results: Phenotypic analysis revealed a significant increase in both body weight (BW) and breast muscle weight with age (p < 0.05). Transcriptomic analysis identified 3521 genes specifically expressed at the age of one day compared with the other 3 ages. These were significantly enriched in pathways related to ribosomal biosynthesis, cytoskeletal regulation, and cell proliferation (p < 0.05). Turquoise and black modules were identified by WGCNA, containing 1563 hub genes, which significantly correlated with BW. Integration of differentially expressed genes and STEM analysis selected 26 BW-related key genes closely associated with muscle growth, including calmodulin 2 (CALM2), heat shock protein 90 alpha family class A member 1 (HSP90AA1), and cholinergic receptor nicotinic delta subunit (CHRND). Protein–protein interaction analysis revealed two functional networks centered around these genes. Enrichment analysis of the STEM profiles indicated that upregulated genes were significantly enriched in autophagy and the ErbB, FoxO, mTOR, and insulin signaling pathways, while genes related to the ribosome, cell cycle, and PPAR signaling pathways were downregulated. Conclusions: This study identified BW-related key genes and pathways, enriching our knowledge of the functional maintenance of chicken BW. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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