Current Issues in Molecular Biology最新文献

Intrahepatic cholangiocarcinoma (iCCA) is a rare yet aggressive malignancy characterized by late-stage diagnosis and poor prognosis. Recent advancements in multi-omics technologies have significantly enhanced the understanding of the molecular features and complex biological mechanisms of iCCA. These approaches have revealed disease molecular heterogeneity and identified biomarkers linked to disease progression, patient prognosis, and therapeutic outcomes. This review synthesizes findings from multi-omics studies, highlighting their clinical implications and advancing the application of precision oncology in iCCA management.

Reversine is a small-molecule Aurora kinase inhibitor known for its pro-apoptotic effects and potential to remodel chromatin architecture. Although its impact on mitotic regulation is established, its effects on telomere dynamics and nuclear organization in chronic myeloid leukemia (CML) remain unclear. This study aimed to investigate the effects of reversine on telomere architecture, genomic instability, and apoptosis in CML cell lines (K-562 and MEG-01). Reversine was applied at increasing concentrations, and cytotoxicity was assessed using caspase-3/7 activation assays. Quantitative PCR was used to measure AURKA and AURKB mRNA expressions. Three-dimensional telomere architecture was analyzed with TeloView^® v1.03 software after Q-FISH labeling to quantify telomere number, signal intensity, aggregation, nuclear volume, and a/c ratio. Reversine induced a dose- and time-dependent apoptotic response in both cell lines and significantly downregulated AURKA and AURKB expressions. Three-dimensional telomere analysis revealed a marked reduction in telomere number and aggregates, signal intensity, and nuclear volume. While reduced signal intensity may indicate telomere shortening, the concurrent decrease in aggregation and altered spatial parameters suggests telomeric reorganization rather than progressive instability. These features reflect structural nuclear remodeling and early apoptotic commitment. Differences between K-562 and MEG-01 responses underscore potential heterogeneity in telomere maintenance mechanisms. Reversine modulates genomic stability in CML cells through dual mechanisms involving Aurora kinase inhibition and telomere architecture remodeling. The integration of 3D telomere profiling highlights reversine's potential as a therapeutic agent targeting nuclear disorganization and mitotic dysregulation in leukemia.

Foodborne illnesses associated with Bacillus cereus represent a persistent public health concern. In this study, we described the isolation and characterization of a novel bacteriophage, ΦBc24, from mud samples, which showed lytic activity against foodborne pathogen B. cereus. Transmission electron microscopy revealed that ΦBc24 exhibited a myovirus morphotype. Biological assays demonstrated that its narrow host range was restricted to B. cereus strains and efficient lytic activity, characterized by a latent period of 10 min and a burst size of 40 PFU per infected cell. The phage exhibited high physicochemical stability, tolerating pH values of 2-12, temperatures of 4-50 °C, salinity up to 1 M NaCl, and ultraviolet exposure, while effectively suppressing host bacterial growth for up to six hours. Whole-genome sequencing showed that phage ΦBc24 possessed a double-stranded DNA genome of 160,311 bp, with 39.48% GC content, and 269 predicted open reading frames (ORFs). Remarkably, 11 tRNA genes were identified, whereas no genes associated with lysogeny, virulence, or antimicrobial resistance were detected. Phylogenetic analysis suggested that ΦBc24 belongs to the genus Caeruleovirus, subfamily Bastillevirinae, family Herelleviridae. Taken together, these results highlight the biological robustness and genomic safety of ΦBc24, supporting its potential as a biocontrol candidate against the foodborne pathogen B. cereus.

To obtain stable Chinese olive reference genes, eight genes (RPN2B, PIP1.4, NIFS1, RPS16, At5g12110, HSC-2, ABCG44, LOS1) exhibiting stable expression were identified as candidate reference genes from the transcriptome. The expression stability of these genes was evaluated across 33 Chinese olive fruit samples from different varieties and seven developmental stages. The most stable reference genes were determined through comparisons using ΔCq, geNorm, NormFinder, BestKeeper, and RefFinder. Analysis revealed that RPN2B and NIFS1 were consistently ranked among the most stable genes across the different algorithms and exhibited stable expression. Therefore, they are recommended as suitable reference genes for gene expression studies in Chinese olive fruits across different varieties and developmental stages. The four different methods of reference gene stability analysis were used to identify the most stable reference genes in different varieties and developmental stages of Chinese olive fruits, which can be used as a reference for the selection of reference genes in the subsequent gene expression studies of Chinese olive fruits.

Castration-resistant prostate cancer (CRPC) remains difficult to treat with conventional chemotherapy. We evaluated a stem cell-based enzyme-prodrug strategy using hTERT-immortalized adipose-derived stem cells engineered to express rabbit carboxylesterase 2 (hTERT-ADSC.CE2) in combination with irinotecan (CPT-11). hTERT-ADSC.CE2 cells were generated via lentiviral transduction and confirmed to overexpress CE2. Their tumor-homing capacity toward PC3 prostate cancer cells was assessed, along with prodrug activation, apoptosis induction, and in vivo tumor suppression in a CRPC mouse model. hTERT-ADSC.CE2 cells demonstrated enhanced migration toward PC3 cells and higher expression of tumor-homing factors than the controls. Under CPT-11, they exhibited a strong "suicide" effect and induced selective killing of PC3 cells, with upregulation of BAX and cleaved caspase-3 and downregulation of BCL-2. By day 14, the combination arm showed significantly lower tumor burden than both the control and irinotecan-alone arms (p < 0.05). The pattern is consistent with intratumoral activation and localized SN-38 exposure. hTERT-ADSC.CE2 combined with irinotecan provides potent, tumor-targeted cytotoxicity and markedly suppresses CRPC progression. This cell-mediated prodrug activation system may represent a promising therapeutic approach for advanced prostate cancer.

Nucleic acid aptamers leverage defined tertiary structures for precise molecular recognition, positioning them as transformative biomedical tools. We engineered AP1-F, a G-quadruplex (G4)-structured aptamer that selectively binds membrane-anchored nucleolin (NCL) non-permeabilizing, overcoming a key limitation of conventional probes. Microscale thermophoresis confirmed nanomolar affinity to NCL. By means of rigorous optimization, AP1-F attained a greater than ten-fold fluorescence signal ratio between malignant and normal cells in co-cultures, exceeding the extensively researched AS1411. Dual-channel flow cytometry demonstrated over 98.78% specificity at single-cell resolution within heterogeneous cell populations, owing to AP1-F's unique membrane localization-unlike AS1411's intracellular uptake, which elicited erroneous signals from cytoplasmic NCL. Competitive binding experiments and Laser Confocal Imaging confirmed that AP1-F specifically identifies cancer cells by binding to the NCL recognition site on the membrane. In pathological sections, AP1-F exhibited a 40.5-fold fluorescence intensity ratio between tumor and normal tissue, facilitating accurate tissue-level differentiation. Significantly, it delineated molecular subtypes by associating membrane NCL patterns with morphometric analysis: luminal-like MCF-7 displayed consistent staining in cohesive clusters, whereas basal-like MDA-MB-468 revealed sporadic NCL with irregular outlines-characteristics imperceptible to intracellular-targeted antibodies, thus offering subtype-specific diagnostic insights. This combination biochemical-morphological approach accomplished subtype differentiation with a single-step, non-permeabilized process that maintained lower cytotoxicity and tissue integrity. AP1-F enhances diagnostic accuracy by utilizing spatial confinement to eradicate intracellular interference, connecting molecular specificity to intraoperative margin evaluation or biopsy categorization.

Macellicephaloides Uschakov, 1955 (Annelida: Polynoidae) is a genus of deep-sea polychaetes characterized by a specialized pharynx bearing two pairs of jaws (with the dorsal pair fused) and three pairs of lateral papillae, the middle pair of which is greatly elongated, and remarkable adaptability to diverse deep-sea habitats. Most species in this genus inhabit abyssal depths (>7200 m), with high diversity in western Pacific trenches, while a few occur in relatively shallow habitats such as deep-sea seamounts and hydrothermal vents. This paper presents a new species, Macellicephaloides lingshuiensis sp. nov., found in deep-sea cold seeps in the South China Sea, representing the shallowest distribution record for the genus to date and the first record from cold seep environments. The classification and phylogeny of Macellicephaloides and related genera have long been the subject of debate. A previous study suggested that Macellicephaloides is nested within the Macellicephala clade, but our analyses-based on 13 mitochondrial protein-coding genes, 12S, 16S, 18S, 28S rRNA, and ITS1-ITS2 sequences-tentatively indicate that these two genera form independent evolutionary clades. Additionally, our phylogeny indicates a close evolutionary relationship between deep-sea Macellicephaloides and cave-dwelling polynoids (e.g., Gesiella), highlighting ecological connections between deep-sea and cave habitats. These conclusions are supported by morphological comparisons and genetic distance analyses. Although the subfamily Macellicephalinae is recovered as a monophyletic group, intergeneric phylogenetic relationships within it remain unresolved, highlighting the need for additional data from more species and genera. We amend the generic diagnosis of Macellicephaloides and provide an identification key to all valid species in the genus. This study clarifies the taxonomy and phylogeny of Macellicephaloides and related taxa, emphasizing the importance of continued sampling in understudied deep-sea habitats to enhance our understanding of their biodiversity.

Epidemiological studies have provided evidence to show that the consumption of coffee and green tea has beneficial effects against cancer. Chlorogenic acid (CGA) in coffee and epigallocatechin-3-O-gallate (EGCG) in tea are involved in these effects. Research also suggests that the anticancer effects of coffee and tea may vary depending on the type of cancer, although the reasons for this remain unclear. As bioactive food factors, CGA and EGCG can contribute to epigenetic modification to exert their anticancer activity. One of the anticancer mechanisms is the one associated with reactive oxygen species (ROS). CGA and EGCG possess activities that initiate anticancer pathways by down-regulating ROS and NF-κB, and up-regulating AMPK. CGA and EGCG can regulate non-coding RNAs, including cancer-associated microRNAs. This review provides updated information on how CGA and EGCG exhibit anticancer activity via ROS-dependent anticancer pathways by regulating microRNA expression.

Background: Asthenozoospermia is defined as a condition in which the total motility of sperm in a semen sample is less than 40%. Due to impairing sperm motility, asthenozoospermia was linked to different mitochondrial DNA (mtDNA) alterations. The current study aimed to investigate the relationship between MT-CO2 gene variants and the development of asthenozoospermia and male infertility in the Jordanian population.

Materials and methods: Semen samples were collected from 196 men, including 119 asthenozoospermic (infertile) and 77 normozoospermia (control), from the Royal Jordanian Medical Services in vitro fertilization (IVF) unit. The isolated mitochondrial DNA (mtDNA) was subjected to a polymerase chain reaction to amplify the MT-CO2 gene. Genetic variants were screened using direct Sanger sequencing. Genotypes and allele frequencies between the case and control groups were compared by the chi-square test and Fisher's exact test.

Results: Three novel variants in the MT-CO2 gene were identified in nine asthenozoospermic cases, including two missense variants (m.8222T>A and m.7997G>A) and one synonymous variant (m.7846 A>G). In addition, the current study reported twenty-three known substitutions. In particular, the rs1556423316 T>C variant showed a significant association with asthenozoospermic infertile men in the studied population (p < 0.05).

Conclusion: The detected missense variants in the MT-CO2 gene in asthenozoospermic infertile men underscore the important role of these variants in the development of asthenozoospermic male infertility.

Currently, a major challenge exists in CRISPR-mediated genome editing research in sheep: the low efficiency of exogenous large DNA fragment targeted integration without drug selection or fluorescence enrichment. This restriction significantly impedes the use of precise genome editing in sheep for agricultural, biological, and biomedical purposes. In this study, we employed the strategy of increasing the local concentration of the homologous repair template at the site of the DNA double-strand break (DSB). We achieved this by fusing the DNA binding domain (BD) of the Gal4 protein (Gal4-BD) to the N-terminal end of the SpCas9 protein using a 32-amino acid (aa) flexible linker. Additionally, we incorporated a 17 bp UAS at the 3' end of the donor template, which can be specifically recognized and bound by Gal-BD. As a result, we observed a significant improvement in the knock-in efficiency of the exogenous large DNA fragment (2997 bp) in sheep fetal fibroblasts (SFFs), increasing it from 5.30% (8/151) to 16.67% (32/192), providing a comparatively efficient and user-friendly method to promote CRISPR-mediated gene knock-in in sheep.