Zhejiang da xue xue bao. Yi xue ban = Journal of Zhejiang University. Medical sciences最新文献

Ovarian tissue cryopreservation and transplantation is currently the only feasible method for preserving both fertility and ovarian endocrine function in prepubertal females. It is indicated for those requiring gonadotoxic therapies such as chemotherapy, radiotherapy, or bone marrow transplantation for malignant diseases, as well as for non-malignant diseases, including immunologic, metabolic, and hematologic benign diseases requiring bone marrow transplantation, and other populations at high risk of premature ovarian insufficiency. The procedure involves laparoscopic retrieval of ovarian tissue, followed by slow-programmed cryopreservation. When the primary disease is cured and fertility or hormonal function restoration is desired, the tissue is thawed and transplanted, most commonly to an orthotopic site. For patients at high risk of ovarian malignancy, pre-transplantation assessment of minimal residual disease in the ovarian cortex is performed using histopathology and molecular biology techniques. Globally, while ovarian tissue cryopreservation and transplantation has led to over 300 live births, the majority result from tissue cryopreserved after puberty. Successful restoration of puberty and subsequent live births following transplantation of tissue frozen before puberty, although demonstrated in reported cases, remain less common. This review systematically summarizes recent advances in the indications, current application status, timing and strategies of ovarian tissue cryopreservation and transplantation, risk assessment of tumor cell reintroduction, and clinical outcomes in prepubertal patients. It also discusses the potential value and current challenges of combining this approach with invitro oocyte maturation techniques, aiming to provide practical references for clinical practice.

Objectives: To investigate the effects of different filtration fractions (FFs) during continuous venovenous hemodiafiltration (CVVHDF) post-dilution.

Methods: This study employed a single-blind, head-to-head randomized controlled design. Patients who underwent daytime continuous renal replacement therapy (CRRT) in the Department of Nephrology, the Second Affiliated Hospital of Nanchang University between April 2022 and June 2023 were prospectively enrolled. They were randomly assigned to either a low FF group (FF set at 20%-<25%) or a high FF group (FF set at 25%-30%). All patients received post-dilution CVVHDF with systemic heparin anticoagulation. The primary outcome was extracorporeal circuit coagulation, comprehensively assessed through dynamic monitoring of arterial pressure, venous pressure, and transmembrane pressure, combined with filter clotting grading at the end of the session. Secondary outcomes included changes in serum creatinine, urea, potassium, and pH levels before and after treatment to evaluate efficacy.

Results: A total of 40 patients were included in both the low FF group and the high FF group. The baseline characteristics showed no statistically significant differences between the two groups (all P>0.05). All patients completed the treatment successfully, with a treatment duration of 10-12 h, and no filters required replacement during the sessions. The differences in arterial pressure, venous pressure, and transmembrane pressure at 2 h, 6 h, and the end of treatment compared to values at 1 h showed no statistically significant differences between the groups (all P>0.05). Furthermore, no significant differences were found in filter clotting grades (including grade Ⅰ and grade Ⅱ clotting) at the end of treatment between the two groups (both P>0.05). The creatinine clearance was significantly higher in the high FF group compared to the low FF group (P<0.01). However, the changes in blood urea nitrogen, serum potassium and pH levels before and after treatment showed no statistically significant differences between the groups (all P>0.05).

Conclusions: For patients with a relatively short treatment duration of 10-12 h undergoing post-dilution CVVHDF, employing an FF of 25%-30% does not pose a higher risk of extracorporeal circuit coagulation compared to an FF of 20%-<25%, but shows higher creatinine clearance.

Breast cancer patients in China tend to be diagnosed at a younger age, making fertility issues a significant clinical and societal challenge. Current evidence indicates that the fertility rate among breast cancer survivors is substantially lower than that of the general population of the same age. Both the disease itself and antitumor treatments-including chemotherapy, radiotherapy, endocrine therapy, targeted therapy, and immunotherapy-can adversely affect female fertility. Therefore, fertility considera-tions should be integrated into the comprehensive management of breast cancer from the time of diagnosis. Several guidelines and consensus statements have been established to direct fertility management in breast cancer patients. Clinical practice has achieved some success in fertility preservation through pharmacological, surgical, and assisted reproductive technologies, which help to mitigate treatment-related damage to fertility. Nevertheless, further progress relies on multidisciplinary collaboration, particularly in addressing the ethical and legal aspects of fertility preservation. Recent advances in research on hereditary breast cancer, risk assessment, and preimplantation genetic testing for polygenic diseases offer new perspectives and directions for fertility management in breast cancer patients. This review systematically summarizes the current fertility status, existing management strategies, and cutting-edge research related to healthy reproduction in breast cancer patients, with the aim of supporting the standardization of fertility management protocols.

Objectives: To investigate the molecular mechanism underlying Leber hereditary optic neuropathy (LHON) caused by the m.3472T>C (p.Phe56Leu) mutation.

Methods: Three large pedigrees (WZL122, WZ676, WZ706) carrying the m.3472T>C mutation were identified and selected from a cohort of 1397 LHON patients. Immortalized lymphoblast cell lines were established from peripheral blood samples obtained from two affected individuals carrying homoplasmic m.3472T>C mutation (derived from one Han Chinese LHON pedigree) and two genetically unrelated, matched healthy controls. The MitoTool software was used to analyze the complete mitochondrial DNA sequence to determine the haplogroup background, and the conservation of the mitochondrial DNA (mtDNA) mutation site was evaluated using the MitoMap database. The secondary structure of the NADH dehydrogenase subunit 1 (ND1) protein was predicted and analyzed using online prediction software. The three-dimensional structure and molecular interaction changes of wild-type and mutant ND1 proteins were visualized and analyzed using PyMOL. Western blotting was performed to determine the expression levels of key proteins. Cellular adenosine triphosphate (ATP) levels were measured using a chemiluminescence assay. Mitochondrial membrane potential and intracellular reactive oxygen species (ROS) levels were assessed by flow cytometry.

Results: mtDNA haplotype analysis showed that all the subjects belonged to the East Asian mitochondrial haplogroup D4. The m.3472T>C mutation results in the substitution of a highly conserved phenylalanine with leucine (p.Phe56Leu) in the ND1 of mitochondrial complex Ⅰ, which may alter the structure and function of ND1. In lymphoblast cell lines carrying the m.3472T>C mutation, manifestations of mutant ND1 instability were observed, including reduced ND1 protein levels, diminished mitochondrial ATP production, decreased mitochondrial membrane potential, and increased mitochondrial ROS production. Concurrently, the expression levels of antioxidant enzymes catalase and superoxide dismutase 2 were elevated (all P<0.01). These findings collectively suggest impaired complex Ⅰ function.

Conclusions: The m.3472T>C mutation contributes to the pathogenesis of LHON by disrupting the structural stability of ND1 in complex Ⅰ, leading to mitochondrial dysfunction.

Premature ovarian insufficiency (POI), characterized by the decline of ovarian function before age 40, significantly compromises fertility and long-term health of patients. Stem cell therapy has emerged as a promising approach for POI. This review synthesizes clinical evidence from studies utilizing cells sourced from adult tissues (e.g., adipose-derived mesenchymal stem cells, bone marrow mesenchymal stem cells and peripheral blood stem cells) as well as from perinatal tissues (e.g., human amniotic epithelial cells and umbilical cord-derived mesenchymal stem cells). Evidence suggests that stem cell transplantation can improve ovarian reserve, reflected by reduced follicle-stimulating hormone levels and increased estradiol and anti-Müllerian hormone levels, with some patients resuming menstruation and achieving pregnancy. However, treatment efficacy is influenced by patient-specific factors and clinical protocols. Optimizing stem cell transplantation protocols is pivotal for enhancing the clinical efficacy and safety. This article elaborates on key optimization strategies, including transplantation timing, delivery routes, and combination therapies, proposing that early intervention and person-alized regimens may improve outcomes. We also discuss patient benefits (such as pregnancy outcomes and quality of life) as well as treatment safety, and explore the clinical value of stem cell therapy for POI. Future research should focus on refining personalized strategies, investigating the therapeutic potential of stem cell-derived agents, and establishing long-term follow-up, thereby advancing POI therapy toward precision medicine and standardized application.

Objectives: To investigate the molecular mechanism by which He's Yangchao recipe improves ovarian function in premature ovarian insufficiency (POI) mice through intestinal flora modulation.

Methods: Forty female ICR mice (aged 6-8 weeks) were intraperitoneally injected with cyclophosphamide (150 mg/kg) to establish a POI model, while 10 untreated mice served as the blank control. The cyclophosphamide-injected mice were randomly divided into four groups with 10 mice in each group: low-dose He's Yangchao recipe (crude herb, 6 g/kg), high-dose He's Yangchao recipe (crude herb, 25 g/kg), estradiol (positive control), and model control (distilled water). Treatments were administered daily by gavage for 6 weeks. Vaginal exfoliated cells were stained with Wright-Giemsa solution to monitor estrous cycles. Serum estradiol and follicle-stimulating hormone (FSH) levels were measured by enzyme-linked immunosorbent assay. Ovarian FSH receptor (FSHR) expression was assessed by immunohistochemistry. Fecal samples from blank control, model control, and high-dose groups underwent metagenomic sequencing to evaluate intestinal microbiota diversity and composition. Glycolysis-related proteins pyruvate kinase M2 (PKM2) and glucose transporter 4 (GLUT4) were analyzed by Western blotting and immunofluorescence.

Results: He's Yangchao recipe restored estrous cyclicity, increased serum estradiol (P<0.05), decreased serum FSH (P<0.05), and upregulated FSHR expression in granulosa cells (P<0.05). Metagenomic analysis revealed significant structural differences in intestinal flora among blank control, model control, and high-dose groups (P<0.05). The high-dose group showed reduced abundance of opportunistic pathogens (e.g., Alistipes, Prevotella, Odoribacter, Blautia, Rikenella) compared to the model control (all P<0.05). Functional enrichment analysis indicated involvement of glycolysis-related pathways. In the model control, PKM2 expression was downregulated (P<0.05), and GLUT4 expression showed a decreasing trend. Both of them were upregulated in the high-dose He's Yangchao recipe group (both P<0.05).

Conclusions: He's Yangchao recipe ameliorates POI in mice by remodeling intestinal flora structure, enhancing glycolytic activity, improving ovarian sex hormone secretion, increasing granulosa cell FSHR expression, and restoring estrous cyclicity.

Primary cilia are microtubule-based organelles that project from the cell surface. They are present in cells from single-celled eukaryotes to vertebrates, including humans. Recent studies have found that primary cilia are also widely distributed in multiple organs and tissues of the reproductive system, where they influence reproductive function by directly participating in or indirectly regulating related signaling pathways, thereby affecting fertility. Primary cilia participate in the regulation of oocyte meiosis and development. They also influence sperm maturation by regulating the homeostatic microenvironment required for spermiogenesis. By mediating Hedgehog (Hh) and Wnt signaling pathways, primary cilia regulate endometrial receptivity and decidual response, thereby influencing the embryo implantation rate. Furthermore, primary cilia control migration, invasion, differentiation, and vascular remodeling of human chorionic villi mesenchymal stromal cells and trophoblasts. Structural or functional impairment of primary cilia may disrupt placental vascular remodeling, leading to placental hypoplasia, potentially through the downregulation of downstream target genes of the Hh signaling pathway. Moreover, primary cilia may be involved in ovarian aging, ovulation, and endocrine function. This article reviews the research progress on the relationship between primary cilia and fertility, explores the potential mechanisms underlying the roles of primary cilia in gamete development, endometrial receptivity, decidualization, placental development, and ovarian reproductive endocrine function, aiming to provide new insights for fertility preservation and the prevention and treatment of human reproductive disorders.

Objectives: To investigate the role of PTEN-induced putative kinase 1 (PINK1) in regulating the viability, migration, and apoptosis of colorectal cancer (CRC) cells, and to explore its potential epigenetic mechanisms.

Methods: PINK1 was overex-pressed or knocked down in HCT116 and DLD1 CRC cell lines using lentiviral vectors, with efficiency verified by qRT-PCR and Western blotting. Cell proliferation was assessed using CCK-8 and colony formation. Cell migration was detected using wound healing and Transwell assays. Apoptosis was assessed using Hoechst 33258 staining. Protein levels of apoptosis-related and histone modification-related markers were analyzed by Western blotting. Genome-wide chromatin accessibility was profiled using assay for transposase-accessible chromatin with sequencing (ATAC-seq).

Results: PINK1 expression was significantly downregulated at both mRNA and protein levels in CRC tissues com-pared to normal tissues. PINK1 overexpression inhibited cell proliferation, colony formation, and migration in HCT116 and DLD1 cells (all P<0.05), whereas PINK1 knockdown promoted these malignant phenotypes (all P<0.05). PINK1 overexpression induced apoptosis, associated with decreased levels of anti-apoptotic proteins (MCL-1, BCL-2, BCL-XL) and increased pro-apoptotic BAX (all P<0.05), without altering p53 expression. Mechanistically, PINK1 overexpression reduced histone H3 lysine 9 trimethylation (H3K9me3) and histone H3 lysine 27 trimethylation (H3K27me3), and increased histone H3 lysine 9 acetylation (H3K9ac) and histone H3 lysine 27 acetylation (H3K27ac). It also downregulated key histone-modifying enzymes, including enhancer of Zeste homolog (EZH)2, EZH1, SUZ12, and histone deacetylase 3 (HDAC3) (all P<0.01). ATAC-seq revealed that PINK1 overexpression increased chromatin accessibility, particularly around transcription start sites.

Conclusions: PINK1 acts as a tumor suppressor in colorectal cancer by inhibiting proliferation and migration, promoting apoptosis, and remodeling the epigenetic landscape through altering histone modifications and enhancing chromatin accessibility.