Isoprothiolane is a widely used organosulfur fungicide detected in surface waters worldwide at concentrations ranging from nanograms to micrograms per liter. However, its toxicological effects on vertebrates remain poorly understood. In this study, we used zebrafish to assess the developmental toxicity of isoprothiolane. Transgenic zebrafish lines Tg(myl7:GFP) and Tg(fli1:GFP) were exposed to isoprothiolane from 6 to 72 h post-fertilization (hpf). Early cardiovascular development and damage were assessed using H&E staining, immunofluorescence staining, and Western blot, while behavioral analysis evaluated locomotor activity. We observed dose-dependent cardiovascular toxicity, including pericardial edema, cardiac linearization, abnormal heart rate, reduced cardiomyocyte numbers, and impaired vascular branching, accompanied by inhibited cell proliferation and increased apoptosis. Transcriptomic analysis identified Wnt pathway suppression as a key mechanism underlying these defects. Furthermore, treatment with BML-284, a Wnt pathway agonist, partially rescued the cardiovascular defects and developmental abnormalities in zebrafish. Therefore, these findings reveal that isoprothiolane disrupts cardiovascular development through Wnt signaling inhibition, providing novel insights into fungicide-induced developmental toxicity and informing ecological risk assessment of organosulfur pesticides.

Pyrethroid pesticides are extensively applied in agriculture and public health, leading to considerable environmental contamination and human exposure. D-tetramethrin is a widely used household insecticide frequently encountered by humans. Although considered low-toxicity, its potential to induce neurotoxicity and motor dysfunction remains poorly defined. Here, we evaluated the developmental neurotoxicity of D-tetramethrin using zebrafish embryos and human SH-SY5Y neuroblastoma cells. In vivo, exposure caused impaired development with increased malformations and reduced locomotor activity. Neurobehavioral testing revealed weakened touch and light-dark responses, anxiety-like behavior, and neuronal abnormalities, indicative of developmental neurotoxicity. Biochemical analyses demonstrated decreased ATPase and AChE activities, reduced GABA levels, compromised antioxidant defenses (SOD, CAT, GSH), and elevated ROS and MDA, consistent with oxidative stress. Transcriptomic profiling identified differentially expressed genes enriched in pathways related to neurodevelopment, neurotransmission, apoptosis, and oxidative stress. To further validate these findings, SH-SY5Y cells were exposed to D-tetramethrin and showed concentration-dependent cytotoxicity, ROS accumulation, and apoptosis, consistent with transcriptomic predictions. In summary, D-tetramethrin disrupts neurodevelopment, neurotransmitter balance, and antioxidant defenses while promoting oxidative stress and apoptosis, leading to neurotoxicity and locomotor dysfunction in zebrafish embryos, further supported by in vitro validation.

Leber congenital amaurosis (LCA) is a severe early-onset inherited retinal dystrophy (IRD). Biallelic variants in the retinoid isomerohydrolase gene RPE65, which encodes a key isomerohydrolase in the visual cycle, represent an important cause of LCA and a molecular target for approved ocular gene therapy. Given the marked ethnic differences in the RPE65 variant spectrum, defining these variants in Chinese patients is important for improving molecular diagnosis and informing subsequent clinical evaluation. Unrelated Chinese families with clinically diagnosed LCA underwent clinical characterization and whole-exome sequencing (WES), and candidate RPE65 variants were validated by Sanger sequencing. Variant pathogenicity was evaluated according to the ACMG/AMP guidelines. Evolutionary conservation was assessed by cross-species sequence alignment, and the structural effects of two rare missense variants were investigated using three-dimensional protein modeling. To characterize the distribution of RPE65 variants in Chinese patients, the present cohort was further integrated with previously reported Chinese LCA cases. Nine distinct RPE65 variants were identified in Chinese patients with LCA, including four missense, two frameshift, two splice-site, and one nonsense variant. All probands showed early-onset severe visual impairment and extinguished ERG responses. Two rare missense variants, p.Thr147Lys and p.His182Asp, lacked prior disease-specific annotation and were supported for classification toward likely pathogenicity. Structural modeling suggested that both variants may affect RPE65 function by altering local structural stability and the substrate-binding environment. Integrated analysis with previously reported Chinese cases showed an observed exon-level distribution of RPE65 variants, with variants more frequently observed in exons 3 and 4, followed by exons 5, 9, and 10. This study expands the RPE65 variant spectrum in Chinese patients with LCA. Integrating genetic and structural evidence improves the interpretation of rare variants. These findings may also support more precise molecular diagnosis and referral for comprehensive evaluation of RPE65-targeted gene therapy eligibility.

Bexarotene is a retinoid X receptor (RXR) agonist that plays a crucial role in cell growth and differentiation. It has shown potential in treating both early- and late-stage cutaneous T-cell lymphoma (CTCL). However, the impact of Bexarotene on the neurodevelopment of aquatic organisms, particularly aquatic vertebrates, remains poorly understood. This study aimed to investigate the effects of various concentrations of bexarotene (3 μg/L, 6 μg/L, and 9 μg/L) on the development of the zebrafish embryonic nervous system, using zebrafish as a model organism. The underlying molecular mechanisms were explored through a combination of pharmacological interventions, molecular biology, histopathology, and transcriptomics. Studies have shown that zebrafish embryos exposed to Bex show significant changes in development, including morphological abnormalities, head malformations, significantly shortened head length and width, reduced fluorescent area, cell apoptosis, shortened spinal motor neuron axon length, abnormal myelin development, decreased oligodendrocytes, cerebellar developmental damage, and abnormal behavior. Transcriptomics and qPCR results showed abnormal expression of neurodevelopmental genes (olig2, mbpa, atoh1a, gfap, ngn1, gap43, etc.). In addition, exposure to medium and high concentrations of bexarotene significantly increased acetylcholinesterase (AChE) activity. Bexarotene activates the Wnt signaling pathway, and treatment with the Wnt inhibitor IWR-1 can partially rescue the neurodevelopmental impairments in embryos. In summary, bexarotene offers new insights into the potential neurodevelopmental risks in zebrafish embryos, emphasizing the importance of preventing drug side effects and ensuring the safe and rational use of medications to protect the health of living organisms.

BACKGROUND:Cordycepin, a major bioactive constituent of Cordyceps militaris, exhibits diverse pharmacological properties including anti-inflammatory and antioxidant activities. However, its neuroprotective effects against aluminum-induced neurotoxicity in aquatic organism models remain largely unexplored. METHODS:In this study, we investigated the neuroprotective effects and intrinsic regulatory pathways by which cordycepin alleviates aluminium chloride (AlCl3)-induced neurotoxicity in zebrafish (Danio rerio) embryos using behavioral, molecular, and transcriptomic approaches. RESULTS:Exposure to low-dose AlCl3 significantly induced developmental neurotoxicity, as manifested by reduced body length and reduced heart rate. AlCl3-treated larvae displayed locomotor deficits and cognitive dysfunction. Notably, cordycepin treatment markedly attenuated these AlCl3-induced neurodevelopmental abnormalities and aluminum chloride (AlCl3)-induced locomotor and anxiety-related behavioral impairments.RNA-Seq analysis revealed 605 upregulated and 241 downregulated genes following co-exposure to cordycepin and AlCl3. Functional enrichment analyses based on Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) screened out defense reaction and inflammatory signaling cascades as the remarkably enriched functional pathways, which implied these biological processes exert key functions in the nerve protection produced by cordycepin. Furthermore, pharmacological inhibition targeting the Wnt cascade via IWR-1-endo (IWR-1) markedly counteracted cordycepin's protective capacity to reverse AlCl3-triggered behavioral impairments, indicating that Wnt signaling is essential for its neuroprotective action. CONCLUSION:Collectively, these findings demonstrate that cordycepin effectively mitigates AlCl3-induced neurotoxicity and AlCl3-induced locomotor and anxiety-related behavioral impairments in zebrafish by regulating inflammatory reactions and the Wnt signal cascade, highlighting its promising value for the clinical intervention of AD.

Acute colorectal obstruction constitutes a surgical emergency in patients with colorectal cancer (CRC) and is associated with high morbidity and mortality when managed with emergency surgery. Although self-expandable metal stents (SEMS) may optimize surgical conditions, concerns remain regarding their long-term oncologic safety. This retrospective cohort study included 355 patients with obstructive CRC treated between 2014 and 2023, including 65 patients in the SEMS group and 290 in the surgery-alone group. Perioperative outcomes, postoperative complications, disease-free survival (DFS), and overall survival (OS) were compared. Propensity score matching and multivariate Cox regression were applied to reduce selection bias and adjust for confounding factors. Patients in the SEMS group had a markedly higher rate of laparoscopic surgery (89.2

With the widespread application of pesticides, water pollution problems are becoming more and more serious, which is very likely to cause harm to fish. Lower vertebrates, including fish, have the ability to repair damaged tissues. The spread of pesticides in the water may affect their regeneration process after injury, leading to their death, thereby affecting the survival rate of the population. Therefore, we used zebrafish as a model animal to evaluate the effect of the pesticide pyrazosulfuron-ethyl on caudal fin regeneration in zebrafish larvae. We exposed zebrafish larvae to 0, 5, 15, and 25 mg/L pyrazosulfuron-ethyl at 3 days after caudal fin amputation. It was found that exposure to pyrazosulfuron-ethyl significantly inhibited caudal fin regeneration and affected the behavior of zebrafish larvae. After exposure to pyrazosulfuron-ethyl, proliferating cells decreased and apoptotic cells increased in the caudal fin of zebrafish larvae. Pyrazosulfuron-ethyl exposure resulted in the decreased number of neutrophils and macrophages, and the downregulation of immune related gene expression levels during caudal fin. Using LPS to activate inflammation can effectively rescue the fin regeneration defects induced by pyrazosulfuron-ethyl. However, inhibiting the Notch signaling pathway and inhibiting reactive oxygen cannot rescue the fin regeneration defects induced by pyrazosulfuron-ethyl. Our results indicate that pyrazosulfuronethyl can inhibit zebrafish caudal fin regeneration by reducing the number of innate immune cells and affecting the normal process of inflammation, thereby inhibiting caudal fin regeneration. This study expands our understanding of the potential effects of the pesticide pyrazosulfuron-ethyl on injured fish, highlights the link between the immune system and the regeneration process, and demonstrates the potential application of fin regeneration in risk assessments of environmental toxicology to assess drug toxicity.

Bexarotene (Bex) is a selective retinoid X receptor (RXR) agonist and is commonly used as an anti-tumor drug in the clinic to treat patients with cutaneous T-cell lymphoma (CTCL). With the widespread use of this drug, people are increasingly concerned about its side effects and safety of use. At present, the effects of bexarotene drugs on the health of organisms remain uncertain, but retinoid drugs are generally biologically active and may pose potential risks to them. Therefore, in this study, we used a zebrafish model to evaluate the effects of Bex on embryonic development. Six hours after fertilization, we exposed zebrafish embryos to 3 μg/L, 6 μg/L, and 9 μg/L bexarotene. At 96 hpf, compared with the control group, zebrafish embryos exposed to bexarotene showed obvious heart and liver development defects, including reduced hatching rate, pericardial enlargement, heart rate disorder, yolk sac edema, small liver area and abnormal photo-optical motor responses. Transcriptome and qPCR results showed abnormal expression of genes related to heart and liver development was induced by Bexarotene. Mechanistically, bexarotene induced a significant upregulation of the transcriptional expression levels of genes related to the Wnt signaling pathway, and IWR-1 was able to effectively rescue the heart and liver developmental defects of zebrafish caused by bexarotene. Therefore, our study showed that bexarotene may cause zebrafish embryonic developmental defects by upregulating the Wnt signaling pathway, revealing the side effects and associated novel mechanisms of bexarotene, and providing a theoretical basis for its safe and effective use in the treatment of clinically related diseases.

BackgroundAmong the disease-causing genes associated with X-linked intellectual disability (XLID), KDM5C is one of the most frequently mutated ones. KDM5C is a widely expressed gene that is most highly expressed in the brain. KDM5C modulates the transcriptional activity of genes through demethylation of H3K4, thereby regulating neural development and normal function. We identified a gene from a Chinese family and found that a nonsense mutation of KDM5C was co-segregated with the intellectual disability (ID). MethodsThe candidate mutant genes of patients with ID phenotype were screened by Whole Exome Sequencing (WES), and DNA Sanger sequencing was performed for genetic analysis. Pathogenicity prediction tools were used to evaluate the pathogenicity of new mutations. The fusion plasmid was constructed and transfected into the cells, and the changes of mRNA and protein levels of the mutants were detected by semi-qRT-PCR and Western Blot, and the subcellular localization changes of mutant proteins were detected by Immunofluorescence technique. ResultThe nonsense mutation in KDM5C (c.2785 C > T, p. R929X) was identified by whole exome sequencing (WES) and confirmed by Sanger sequencing, resulting in a truncated protein. The mutation was determined by pathogenicity prediction tool able to find non-sense mediated mRNA decay (NMD). Semi-qRT-PCR and Western Blot showed that the mRNA levels of the mutant gene were down-regulated, while the protein level was up-regulated. Additionally, the subcellular localization of the mutant protein changed. ConclusionsThe KDM5C mutation found in our study leads to changes in protein levels through NMD and/or protein degradation, and produces residues lacking nuclear localization, thus altering the subcellular localization of the protein. These results may lead to changes in the expression of KDM5C target genes, ultimately contributing to the clinical phenotype observed in the patients.

Dementia with Lewy bodies (DLB) is a significant cause of dementia. However, the limited availability of animal and cellular models that accurately replicate early DLB pathogenesis hampers the understanding of how Aβ plaques influence α‐synuclein (αSyn) pathologies. This study addresses this gap by co‐culturing primary neurons with adult hippocampal brain slices from either wild‐type or Alzheimer's disease (AD) mice containing abundant Aβ plaques and cytokines. Neurons exposed to AD slices showed impaired dynein‐dependent organelle trafficking, reducing endosome–lysosome fusion and causing defective degradation of amyloidogenic αSyn fibrils, thus increasing αSyn inclusions. Notably, an abnormal pre‐accumulation of dynein in AD mice suggests that dysfunctional dynein may serve as a nucleation site for αSyn aggregation upon exposure to pathogenic fibrils. Furthermore, Rab7 activation successfully restored endo‐lysosomal degradation of αSyn fibrils and reduced inclusion formation in mouse models presenting with both Lewy body and Aβ pathologies. These results highlight the dynein‐dependent endo‐lysosomal pathway as a promising therapeutic target for mitigating αSyn‐related pathologies in co‐existing Aβ burden, characteristic of many DLB cases.

Hinokitiol is a natural compound collected from the trunk of cypress, belonging to the tropolone family of compounds. It has anti-inflammatory, anti-tumour and antibacterial activities, making it a natural product with a wide range of applications. It is used as an additive in hair growth agents, toothpaste, make-up and furniture wood. However, the toxicity of hinokitiol remains poorly understood. Therefore, the cardiovascular and developmental toxicity of hinokitiol in organisms was investigated in zebrafish larvae. In the study, zebrafish embryos were exposed to hinokitiol for 3 days to study the developmental and cardiovascular toxicity of hinokitiol. Concentrations for the hinokitiol toxicity test were set at 0, 0.4, 0.8, 1.2, 1.6, 2.0 and 2.2 mg/L. The experimental concentrations of 0, 0.8, 1.2 and 1.6 mg/L were subsequently determined based on phenotype. The results showed that exposure to hinokitiol resulted in increased mortality, changes in hatching rates and abnormalities in the apparent morphology of zebrafish embryos/larvae (shortened body length, reduced eye area, pericardial edema, and abnormal heart rate). In addition, hinokitiol impaired the cardiovascular system of zebrafish larvae, as evidenced by the absence of morphological features of the atria and ventricles, linearization of the heart, and reduction in the area and abundance of blood vessels. In addition, hinokitiol affects mitochondrial function by affecting iron ion levels and generates oxidative stress leading to apoptosis and which then generates cardiovascular toxicity. Our findings suggest that hinokitiol causes mitochondrial dysfunction and oxidative stress through iron chelation, which in turn triggers apoptosis, ultimately leading to cardiovascular and developmental toxicity in zebrafish larvae. The study provides new insights into the safety of hinokitiol and natural products.

BACKGROUND:Pontocerebellar hypoplasia type 11 (PCH11) is an autosomal recessive disorder caused by variants in TBC1D23. The molecular basis for its clinical heterogeneity remains poorly understood. Here, we identified a novel TBC1D23 variant in a Chinese family, investigated its underlying pathogenic mechanisms, and systematically reviewed the clinical phenotypes of all reported cases of PCH11. RESULTS:We identified a novel homozygous frameshift variant, c.511_512delTT (p.F171Qfs*8), in TBC1D23. The patient exhibited a severe phenotype, including marked pontocerebellar hypoplasia, a thinned corpus callosum, global developmental delay, and severe language and motor impairments. Mechanistic studies in a zebrafish model revealed that the mutant transcript partially escaped nonsense-mediated decay (NMD), with expression levels at approximately 50% of the wild-type. In vitro, the resultant truncated protein showed enhanced stability and aberrant cytoplasmic distribution instead of its normal Golgi localization. Furthermore, its expression significantly inhibited cell proliferation. CONCLUSION:Our study identifies c.511_512delTT as a novel pathogenic variant in TBC1D23. We propose the severe phenotype stems from a primary loss-of-function (LoF), which is likely exacerbated by the cytotoxic effect of the truncated protein produced via partial NMD escape. Our findings suggest this mutant protein exhibits increased stability. This model provides a novel explanation for the phenotypic heterogeneity in PCH11 and expands the mutational spectrum of this disorder.

Reproductive system diseases have become a major health challenge facing humans, so extensive investigations are needed to understand their complex pathogenesis and summarize effective treatments. In the study of reproductive diseases, mice are the most commonly used animal model. However, the cost and time required to establish mouse animal models are high. The existing zebrafish model can solve this problem well. Zebrafish is an animal model with great application prospects and has lots of advantages, including high degree of genetic conservation with humans, short reproductive cycle, transparent embryos, and rapid growth, providing unique opportunities for high-throughput drug screening and identification of potential treatments. Researchers have successfully used chemical induction, physical damage, gene editing technology, etc., to induce reproductive system damage in zebrafish to study the biological processes related to its reproductive diseases. Therefore, in this review, the main models and related advantages of zebrafish in reproductive diseases are summarized, the pathological mechanisms of zebrafish as a reproductive disease model are clarified, and new perspectives and valuable insights are provided for the treatment of human reproductive diseases. The literature and data cited in the review are all from PubMed, covering important research results on zebrafish reproductive diseases in the past 10 years.

Purpose: To further explore the influence of genotype, including mutation type and structural domain, on the severity of macular atrophy, we measured the central retinal thickness (CRT) in patients with ABCA4-related retinopathy. Methods: A total of 66 patients were included in the cohort. This was a retrospective investigation. The patients were tested using whole exon sequencing and ophthalmic exams, including slip lamp exams, best-corrected visual acuity, optical coherence tomography, fundus photo, and fundus autofluorescence. Results: In this study, we discovered that mutations on nucleotide binding domains (NBD) lead to less CRT (45.00 +/- 25.25 mu m, 95% CI: 31.54-58.46) had significantly less CRT than the others (89.75 +/- 71.17 mu m, 95% CI: 30.25-149.25, p = 0.032), and could accelerate the rate of CRT decrease. Conclusions: Our study provides new perspectives in the understanding of ABCA4-related retinopathy.

The colon is the largest compartment of the immune system, with innate immune cells exposed to antigens in the environment. However, the mechanisms by which the innate immune system is instigated are poorly defined in colorectal cancer (CRC). Here, a population of CD16+ neutrophils that specifically accumulate in CRC tumor tissues by imaging mass cytometry (IMC), immune fluorescence, and flow cytometry, which demonstrated pro-tumor activity by disturbing natural killer (NK) cells are identified. It is found that these CD16+ neutrophils possess abnormal cholesterol accumulation due to activation of the CD16/TAK1/NF-κB axis, which upregulates scavenger receptors for cholesterol intake including CD36 and LRP1. Consequently, these region-specific CD16+ neutrophils not only competitively inhibit cholesterol intake of NK cells, which interrupts NK lipid raft formation and blocks their antitumor signaling but also release neutrophil extracellular traps (NETs) to induce the death of NK cells. Furthermore, CD16-knockout reverses the pro-tumor activity of neutrophils and restored NK cell cytotoxicity. Collectively, the findings suggest that CRC region-specific CD16+ neutrophils can be a diagnostic marker and potential therapeutic target for CRC.

BackgroundThe global increase in the aging population has led to a higher incidence of osteoporosis among the elderly.ObjectiveThis study aimed to evaluate the protective properties of pinoresinol diglucoside (PDG), an active constituent of Eucommia ulmoides, against dexamethasone-induced osteoporosis and chondrodysplasia.MethodsA zebrafish model of osteoporosis was established by exposing larval zebrafish to dexamethasone. The impact of PDG on bone mineralization was assessed through alizarin red and calcein staining. Alkaline phosphatase activity was quantified to evaluate osteoblast function. The influence of PDG on chondrogenesis was estimated using alcian blue staining. Fluorescence imaging and motor behavior analysis were employed to assess the protective effect of PDG on the structure and function of dexamethasone-induced skeletal teratogenesis. qPCR determined the expression of osteogenesis and Wnt signaling-related genes. Molecular docking was used to assess the potential interactions between PDG and Wnt receptors.ResultsPDG significantly increased bone mineralization and corrected spinal curvature and cartilage malformations in the zebrafish model. Furthermore, PDG enhanced swimming abilities compared to the model group. PDG mitigated dexamethasone-induced skeletal abnormalities in zebrafish by upregulating Wnt signaling, showing potential interaction with Wnt receptors FZD2 and FZD5.ConclusionPDG mitigates dexamethasone-induced osteoporosis and chondrodysplasia by promoting bone formation and activating Wnt signaling.

BACKGROUND:Chromosome gains or localized amplifications are frequently observed in human gastric cancer (GC) and are major causes of aberrant oncogene activation. However, the significance of long non-coding RNAs (LncRNAs) in the above process is largely unknown.METHODS:The copy number aberrations (CNAs) data of GC samples were downloaded and analyzed from the TCGA database. qRT-PCR and fluorescence in situ hybridization were used to evaluate the expression of Linc01711 in GC. The effects of Linc01711 on GC progression were investigated through in vitro and in vivo assays. The mechanism of Linc01711 action was explored through transcriptome sequencing, chromatin immunoprecipitation sequencing, RNA immunoprecipitation, RNA pull-down and chromatin isolation by RNA purification (ChIRP) assays.RESULTS:We report for the first time a novel DNA copy number amplification-driven LncRNA on chromosome 20q13, designated Linc01711 in human GC, which is highly associated with malignant features. Functionally, Linc01711 significantly accelerates the proliferation and metastasis of GC. Mechanistically, Linc01711 acts as a modular scaffold to promote the binding of histone acetyltransferase HBO1 and histone demethylase KDM9. By coordinating the localization of the HBO1/KDM9 complex, Linc01711 specifies the histone modification pattern on the target genes, such as LPCAT1, and consequently facilitates the cholesterol synthesis, thereby contributing to tumor progression.CONCLUSIONS:Our findings suggest that copy number amplification-driven Linc01711 may serve as a promising prognostic predictor for GC patients and targeting Linc01711-related cholesterol metabolism pathway may be meaningful in anticancer strategies.

Isavuconazole is a broad-spectrum antifungal drug used for the treatment of serious infections caused by invasive aspergillosis and mucormycosis in adults. With the continuous use of this drug, its safety and environmental impact have received increasing attention. However, information on the adverse effects of the drug is very limited. Fish is a particularly important model for assessing environmental risks. In this study, the aquatic vertebrate zebrafish was used as a model to study the toxic effects and mechanisms of isavuconazole. We exposed zebrafish embryos to 0.25, 0.5, and 1 mg/L of isavuconazole 6 h after fertilization. The results showed that at 72 hpf, isavuconazole exposure reduced heart rate, body length, and survival of zebrafish embryos compared to controls. Secondly, when isavuconazole reached a certain dose level (0.25 mg/L), it caused morphological changes in the Tg(elavl3:eGFP) transgenic fish line, with the head shrunk, the body bent, the fluorescence intensity becoming weaker, the abnormal motor behaviour, etc. At the same time, exposure of zebrafish embryos to isavuconazole downregulated acetylcholinesterase (AchE) and adenosine triphosphate (ATPase) activities but upregulated oxidative stress, thereby disrupting neural development and gene expression of neurotransmitter pathways. In addition, astaxanthin partially rescued the neurodevelopmental defects of zebrafish embryos by downregulating oxidative stress. Thus, our study suggests that isavuconazole exposure may induce neurodevelopment defects and behavioural disturbances in larval zebrafish.