Extracellular vesicles (EVs) are membrane-bound vesicles secreted by cells, including exosomes, microvesicles, and apoptotic bodies, which play critical roles in intercellular communication, material transport, and signal transduction. In recent years, increasing evidence has highlighted the essential function of EVs in early embryo development. By carrying bioactive molecules such as proteins, nucleic acids (e.g., mRNA and miRNA), and lipids, EVs regulate embryonic gene expression, cell proliferation, differ-entiation, and the microenvironment. Studies have shown that EVs derived from various segments of the female reproductive tract can enhance embryonic developmental potential, improve embryo quality, and facilitate implantation. Additionally, EVs secreted by embryos themselves participate in intercellular communication and play pivotal roles during embryogenesis. This review summarizes recent advances in understanding the functions of EVs in early embryo development, discusses their roles in mediating cell-to-cell com-munication and regulating gene expression, and explores the potential applications in reproductive medicine and clinical practice, offering new perspectives for optimizing assisted reproductive technologies.
With the increasing survival rates of cancer patients, the demand for fertility preservation in women has become increasingly prominent. Ovarian tissue cryopreservation and transplantation (OTCT) is an emerging fertility preservation technique that offers a unique advantage over embryo or oocyte cryopreservation, as it does not require ovarian stimulation. This makes it particularly suitable for prepubertal girls requiring urgent gonadotoxic therapy and reproductive-age women who cannot delay cancer treatment. Clinical evidence confirms that OTCT can effectively restore female fertility-especially the potential for natural conception-and restore ovarian endocrine function. The OTCT process involves key steps such as patient evaluation, tissue processing, cryopreservation, and transplantation. The patient's age at cryopreservation, ovarian reserve status, and prior exposure to gonadotoxic therapy significantly influence the outcomes of fertility preserva-tion. Optimal tissue preparation and the choice of cryopreservation method are critical for preserving ovarian tissue viability. During processing, the size of ovarian tissue fragments must be carefully controlled to balance freezing efficiency and post-transplantation viability, with adjustments based on individual patient factors. Slow freezing remains the mainstream clinical method, while vitrification is still considered experimental, with its efficacy and safety under ongoing investigation. The number, size, and transplantation site of ovarian tissue grafts impact their biological activity and functional outcomes. Both orthotopic and heterotopic transplantation can restore endocrine function, but orthotopic sites are superior for restoring fertility. A major safety concern in OTCT is the potential risk of reintroducing malignant or premalignant cells upon reimplantation. Innovative techniques such as in vitro maturation of oocytes and artificial ovaries are being explored to mitigate this risk. This review summarizes recent clinical advances in OTCT, with a focus on its indications, efficacy, implementation strategies, and safety profile, aiming to provide a reference for further research and clinical practice in this field.
Objectives: Ginkgolide B (GB), a natural bioactive compound, acts on multiple molecular targets; however, its potential multi-target-based therapeutic indications remain unexplored. This study used a network medicine framework to systematically predict novel therapeutic indications for GB, aiming to provide evidence to support its clinical repositioning.
Methods: GB targets were comprehensively identified by integrating data from TTD, CTD, and BindingDB databases, followed by literature-based validation. Disease-gene associations were compiled by integrating GWAS and OMIM datasets. The topological proximity between GB targets and disease modules within the human protein-protein interaction network was quantified using network proximity metrics, with statistical significance evaluated via Z-score and permutation testing. Candidate diseases were classified according to MeSH terms, visualized through network mapping, and further characterized by gene set similarity based on the Jaccard index. Experiments in RAW 264.7 macrophages were conducted to validate the inhibitory effect of GB on pro-inflammatory cytokines in cells.
Results: Twelve experimentally validated GB targets were identified, and a disease-gene set encompassing 680 diseases was established. Network proximity analysis revealed significant associations between GB and 22 diseases (Z<0, P<0.05), of which 11 have prior literature support and 11 represent novel predictions. Disease classification indicated that the predicted indications of GB were primarily enriched in immune and inflammatory disease categories, with notable examples including juvenile idiopathic arthritis (Z=－3.10) and Crohn's disease (Z=－3.05). In vitro experiments with macrophages demonstrated that GB suppresses the production of pro-inflammatory cytokines, thereby supporting its potential as a therapeutic agent for immune-mediated inflammatory diseases. Network analysis suggested that GB may exert pleiotropic regulatory effects across multiple diseases via key targets such as MAPK1. The strongest gene set similarity was observed between juvenile idiopathic arthritis and ankylosing spondylitis (Jaccard index=0.60).
Conclusions: This study demonstrates the utility of a network medicine approach in systematically predicting and partially validating novel indications for GB, particularly within immune-inflammatory disorders, including type 1 diabetes, inflammatory bowel disease, and rheumatoid arthritis. These findings provide a theoretical framework for the clinical repositioning of GB and highlight the potential utility of the network-based strategies in the systematic exploration of natural products.
Objectives: To characterize the biological properties of double-negative T (DNT) cells isolated from leukoreduction filter residues.
Methods: Leukoreduction filters containing residues from 400 mL whole blood units (n=6) were collected from a blood center. Filters were back-flushed with normal saline, and the eluate was concentrated to obtain leukoreduction filter residues. Leukocytes in the residues were counted by dual-fluorescence staining. DNT cells were then isolated from the residues using antibody-mediated adsorption and density gradient centrifugation. Both cryopreserved and fresh unstimulated DNT cells derived from the residues were subjected to in vitro culture. Following culture, cells were assessed for expansion fold, viability, immunophenotype, differentiation status, and cytotoxicity against target cells using dual-fluorescence staining and flow cytometry, with comparisons made to DNT cells derived from whole blood.
Results: The leukocyte recovery rate achieved through reverse flushing of the leukocyte reduction filter was (41.9±14.7)%. Compared to whole blood, the DNT cell starting material obtained from filter residues showed no significant difference in total T-cell content (P>0.05). However, the viability and purity of the resulting DNT cell starting materials were significantly lower (both P<0.05). After 17 days of culture, DNT cells from filter residues and whole blood showed no significant differences in expansion fold, immunophenotype, differentiation status, or cytotoxicity toward target cells (all P>0.05). However, the viability of DNT cells from residues was significantly lower than that of whole blood-derived DNT cells [(86.0±4.2)% vs. (92.2±1.2)%, P<0.05]. After thawing (post 3 or 15 days of cryopreservation) and 17 days of culture, DNT cell starting materials from residues showed comparable immunophenotype, expansion fold, and differentiation status to their non-cryopreserved counterparts from the same source (all P>0.05). However, the viability of DNT cells cryopreserved for 3 days [92.4% (91.8%, 92.8%)] and the cytotoxicity against target cells of those cryopreserved for 15 days [91.3% (89.4%, 95.1%)] were significantly higher than those of non-cryopreserved DNT cells [87.8% (82.0%, 89.0%) and 70.9% (67.3%, 80.2%), respectively] (P<0.05).
Conclusions: DNT cells derived from leukoreduction filter residues exhibited highly comparable characteristics to those from whole blood in terms of expansion, purity, differentiation, and biological potency. Furthermore, their biological activity post-cryopreservation and revival remained largely similar to non-cryopreserved cells. These findings suggest that leukoreduction filter residues represent a promising alternative source of starting material for manufacturing off-the-shelf, allogeneic DNT cell therapeutics.
This retracts the article DOI: 10.3724/zdxbyxb-2022-0519.
Complex crown-root fractures in the esthetic zone refer to a type of dental trauma occurring in the anterior region, characterized by concurrent fractures involving both the crown and the root, with associated pulp exposure and periodontal tissue injury. These injuries consistently exhibit critical anatomical features, including a fixed palatal fracture location below the alveolar crest, compromised residual tooth structure, and frequent encroachment of the biological width. To predict treatment outcomes, a risk assessment framework based on the restoration-tooth-periodontium interface was developed. Resistance risk was evaluated by assessing the type of residual dentin ferrule and the length of the root within the alveolar bone, while periodontal risk was assessed according to gingival phenotype and alveolar bone morphology. Based on these risk dimensions and the principles of aesthetics, stability, and minimally invasive treatment, a diagnostic classification system was established to categorize fractures into three types: favorable, intervention and high-risk. Type-specific management strategies were proposed: for favorable cases, crown lengthening combined with deep margin elevation to reduce periodontal risk is recommended; for intervention cases, orthodontic extrusion or surgical extrusion is applied to simultaneously address both ferrule deficiency and biological width violation; for high-risk cases, extraction followed by implant restoration is advised due to limited root preservation value. The presented classification enables clinicians to adopt a scientific and structured approach to treatment planning for these complex crown-root fractures in the aesthetic zone.
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