The grass carp (Ctenopharyngodon idella) aquaculture industry is severely threatened by GCRV-induced hemorrhagic disease. Breeding GCRV-resistant grass carp is an effective strategy to mitigate these losses, but molecular targets with in vivo functional validation remain scarce. In this study, we identified 7-dehydrocholesterol reductase (Dhcr7), a highly conserved enzyme in cholesterol biosynthesis, as a promising target for GCRV-resistant breeding. Dhcr7 is predominantly expressed in the hepatopancreas and brain of grass carp. To assess its breeding potential, we generated dhcr7 knockout grass carp using CRISPR/Cas9 and selected crispants with > 50% mutation rates. Disruption of dhcr7 gene in mutants was confirmed at both mRNA and metabolite levels. Following GCRV-II challenge, these dhcr7 crispants showed ∼40% higher survival rates than wild-type controls, along with significantly upregulated antiviral immune, reduced viral loads, and markedly attenuated hepatopancreas damage. Mechanistically, Dhcr7 deficiency increased Irf3 protein levels, leading to enhanced activation of antiviral immunity. Importantly, dhcr7 disruption did not impair growth performance or muscle morphology. Collectively, our work uncovers a key antiviral role of grass carp dhcr7 and provides a valuable molecular target and a practical strategy for disease-resistant breeding in grass carp.

PURPOSE. Retinitis pigmentosa (RP) is one of the main causes of hereditary blindness, and its genetic mode shows high heterogeneity. Among them, the mutation of the CERKL gene has been identified as the causative gene related to autosomal recessive hereditary RP. The underlying pathogenic mechanisms have remained obscure, hindering the development of effective therapies. This study aimed to elucidate the pathogenic mechanism linking CERKL deficiency to retinal degeneration and to identify a potential mechanism-based therapy. METHODS. We used cerkl-/- zebrafish model, human retinal pigment epithelium (RPE)-1 cells, and utilized integrated multi-omics approaches (metabolomics, transcriptomics, and phosphoproteomics). Key findings were validated through lipid staining, biochemical assays, transmission electron microscopy (TEM), and rescue experiments. RESULTS. CERKL deficiency triggered progressive lipid droplet (LD) accumulation in the RPE, associated with a profound reduction in phosphatidylcholine (PC) levels. Multiomics integration revealed that PC deficiency stemmed from hypophosphorylation of the rate-limiting enzyme phosphate cytidylyltransferase 1A (PCYT1A) at a conserved serine residue (S331). Reconstitution of phosphomimetic PCYT1A (S331D) rescued LD pathology. Critically, exogenous PC supplementation alleviated LD accumulation, preserved photoreceptor outer segment structure, and improved retinal morphology in cerkl-/- zebrafish. CONCLUSIONS. Our work establishes dysregulated PC metabolism due to PCYT1A hypophosphorylation as a pathogenic driver in CERKL-deficient RP. We identify PC supplementation as a readily translatable, metabolic therapy for this genetically defined form of retinal degeneration.

Duhuo Jisheng Decoction (DHJSD) shows promise for treating intervertebral disc degeneration (IVDD), but its mechanisms concerning autophagy and fibrosis are unclear. Using network pharmacology, metabolomics, UHPLC-Q-TOF/MS, and functional studies (in vitro and in vivo), we systematically explored DHJSD's molecular mechanisms. DHJSD has 254 constituents; those may regulate inflammation, apoptosis, and metabolic processes. DHJSD attenuates ECM/fibrosis-related changes, lowers BMP2 expression, is associated with reduced TGF-β/Smad2/3 phosphorylation, and partially improves annulus fibrosus morphology. SB431542 attenuated IL-1β-induced TGF-β pathway activation and BMP2 expression, supporting the involvement of this pathway in DHJSD-related regulation of fibrosis markers. The levels of serum IL-1β and TNF-α significantly decreased in animal models. Through glycerophospholipid and sphingolipid metabolism, DHJSD reshapes lipid homeostasis and may be associated with reduced TGF-β overactivation by downregulating pro-fibrotic compounds and upregulating anti-inflammatory metabolites. DHJSD modulates autophagy-related markers via controlling the LC3-II/LC3-I ratio and BCL2, P62 expression. DHJSD may affect glycolysis-related and oxidative phosphorylation-related changes and may be associated with phosphatidylcholine/ethanolamine-related mitochondrial membrane changes. DHJSD treats IVDD via a "metabolic reprogramming-TGF-β-related regulation-autophagy/mitochondrial-related remodeling" network, suggesting a potential multi-target strategy and demonstrating the value of multi-omics in analyzing traditional medicine.

Background The primary pathogenic mechanism of lower back pain is intervertebral disc degeneration (IVDD), and the phenotypic change of nucleus pulposus cells and matrix degradation are caused by an imbalance in the "inflammation - autophagy - fibrosis" axis. Despite Tong'an decoction's obvious therapeutic benefits, it is unclear if its principal ingredient, quercetin, controls mitochondrial autophagy and postpones IVDD by interfering with TNF signaling. Methods Tong'an decoction's various components were screened using the traditional Chinese medicine database, and quercetin's main targets were found. By combining transcriptome differential analysis and module analysis, IVDD core genes may be identified. These genes can then be intersected with the autophagy genes to identify autophagy-associated IVDD genes. Joint validation of core genes, additional single-cell sequencing study of cell subpopulation dynamics, and external data validation of core genes expression. The in vivo effectiveness of quercetin was confirmed by imaging and pathological tissue staining, acupuncture rat models, the identification of inflammatory markers, and the RNA detection of important genes. The mechanism of action of quercetin was anticipated using a quercetin target interaction network. Finally, the lipopolysaccharide induced nucleus pulposus cells model was used for molecular mechanism and functional validation. Results 677 IVDD related autophagy genes (such as HIF1A, TNF, BCL2, and LC3) were screened. Functional enrichment shows that these genes are significantly involved in mitochondrial autophagy, apoptosis, ferroptosis, and inflammatory signaling pathways such as TNF, MAPK, and NF-KB. External verification found that the levels of inflammatory factors IL-1 β and TNF - α were elevated in IVDD tissues, and key genes for autophagy and apoptosis were expressed. Single cell sequencing detected different states of nucleus pulposus cells, among which fibrous nucleus pulposus cells are an important pathological type in IVDD. Trajectory analysis reveals the transition of nucleus pulposus cells from steady state to fibrotic phenotype, accompanied by the secretion of inflammatory factors by macrophages. The animal model showed that the collagen arrangement in the nucleus pulposus tissue of the model group was disordered, fibrosis occurred, and the expression of inflammation, autophagy, apoptosis hub genes HIF1A, MAPK1, NFKB, CASP3, etc. was upregulated, while BCL2 was downregulated. Cell experiments have confirmed that inflammatory stimulation leads to depolarization of mitochondrial membrane potential, elevation of autophagy markers, and swelling of mitochondrial structure, which were alleviated by quercetin intervention. Conclusion The study found that TNF/IL-1β driven macrophage infiltration stimulates the NF-κB/MAPK pathway, increasing inflammation-induced mitochondrial autophagy dysregulation. Additionally, HIF-1α hypoxic stress accelerates the transition of nucleus pulposus cells into fibroNPCs. Quercetin treatment can drastically reduce TNF signaling, restore mitochondrial autophagy equilibrium, and reverse fibrosis transformation. This study provides a complete proof chain of "components targets phenotype" for treating IVDD with Tong'an Tang, establishing the groundwork for clinical translation.

THOR (testis-associated highly conserved oncogenic long non-coding RNA) is a highly conserved and testis-enriched lncRNA across vertebrates that plays diverse roles in various cancers. However, its physiological function and regulatory mechanism in testes remain largely unknown. Here, we investigated the genomic location and expression pattern of THOR in the model organism zebrafish, and generated a homozygous THOR knockout model using CRISPR-Cas9 technology. Loss of THOR in zebrafish impaired spermatogenesis, leading to oligospermia (38.7% reduction in sperm count), reduced sperm motility, sperm ultrastructural defects, and decreased fertilization rates. RNA-seq analysis of WT and THOR knockout testes revealed dysregulation of cell cycle-related genes, including cdkn1d, foxo1a, tsc1a, tsc2, atrx, and rad21b. RNA pulldown assays in zebrafish testes identified 486 potential THOR-interacting proteins primarily involved in ribosome biogenesis, RNA splicing, chromatin architecture, and meiotic progression. Notably, the core synaptonemal complex components Sycp1, Sycp2, and Sycp3 were all captured as THOR-binding partners. We further demonstrated that THOR directly interacts with Sycp3 and positively regulates its protein levels. Immunostaining assays on chromosome spreads revealed a significantly higher frequency of discontinuous Sycp3 signals in THOR-/- testes, suggesting the presence of meiosis defects caused by Sycp3 downregulation. Our findings expand the understanding of lncRNA-mediated control of spermatogenesis and male infertility by providing the first evidence that lncRNA THOR interacts with the synaptonemal complex to regulate meiosis progression.

Microbial communities are increasingly recognized for their essential roles in the reproductive system. However, the microbial communities in healthy gonads—neither in the ovary nor the testis—have not been extensively explored, particularly with respect to sex differentiation. Sex reversal is a unique mode of sex differentiation that is a well-documented phenomenon in various animal species, with the swamp eel (Monopterus albus) being a notable example of a hermaphroditic species that undergoes natural female-to-male sex reversal. Thus, swamp eel offers a robust system for exploring gonad microbial communities and their biological and functional significance. Our study revealed a living microbial community in the gonads of healthy swamp eel, with microbial loads comparable to those found in three distinct niches: gut, skin, and blood. The gonad microbial communities shared > 55 1. Gonadal microbial communities at homeostasis are partially derived from the gut and blood microbiomes. 2. The dominant ovarian bacteria Bacillus leads to ovarian dysbiosis. 3. Prostaglandin E3 may serve as a metabolic biomarker in response to Bacillus. 4. Bacillus induces testicular inflammation and reduces sperm motility in hermaphroditic swamp eel.

Oncogenic KRAS, a notorious driver of cancer progression, remains a therapeutic challenge. In hepatocellular carcinoma (HCC), KRAS overexpression correlates with tumor aggressiveness. Here, we demonstrate that NSC48160 induces HCC cell death by suppressing KRAS expression. Metabolomic profiling revealed that NSC48160 significantly enhances intracellular tricarboxylic acid (TCA) cycle activity and fructose metabolism, disrupting redox homeostasis, and triggering ferroptosis. Combining NSC48160 with the SLC7A11 inhibitor HG106 synergistically eliminated HCC cells in vitro and suppressed tumor growth in vivo. Mechanistically, NSC48160 indirectly inhibits the Nrf2-SLC7A11-GPX4 axis, as evidenced by ferroptosis-pathway array assays. Specifically, NSC48160 downregulates Nrf2 expression, thereby suppressing its downstream targets GPX4 and SLC7A11, ultimately promoting ferroptosis. Our findings establish NSC48160 as a novel KRAS inhibitor that induces ferroptosis through metabolic and redox reprogramming, offering a promising therapeutic strategy for KRAS-driven HCC.

MicroRNAs are important post-transcriptional regulators, yet the molecular crosstalk between miRNAs and their target genes during sex differentiation remains poorly understood. Medaka (Oryzias latipes), the first fish in which the sex determination gene was identified, serves as an ideal model for studying this process. Here, we generated gonadal RNA-seq and small RNA-seq data from XYDMY- females, wild-type females and males to explore this crosstalk. A total of twenty-seven RNA-seq datasets, comprising 188 Gb of raw reads, and twenty-seven small RNA-seq datasets, totaling 18 Gb of raw reads, were collected, covering 10, 30 and 120 days. After optimizing the mapping and normalizing, we conducted transcriptional and post-transcriptional dynamic analyses of differentially expressed genes and miRNAs between WT females and males, as well as between WT females and XYDMY- females. Additionally, we integrated the RNA-seq and small RNA-seq data to construct comprehensive interaction networks and performed a detailed analysis of the temporal dynamics in gene and miRNA expression. These resources offer valuable insights into the transcriptional regulation of gonadal differentiation and development in vertebrates.

Immune checkpoint inhibitors (ICIs) targeting programmed cell death protein 1 (PD-1) or programmed death ligand 1(PD-L1) respond well to deficient-microsatellite(dMMR) colorectal cancer and poorly to proficient-microsatellite (pMMR) CRC. Anti-vascular therapy is the standard backline treatment regimen for advanced metastatic colorectal cancer and also potentiates the immunotherapeutic efficacy of CRC by promoting immune cell infiltration and remodeling the tumor immune microenvironment (TIME). However, it is not clear whether combining radiotherapy, anti-vascular and anti-PD-1 can affect the efficacy of pMMR CRC. In this experiment, we investigated the antitumor efficacy of radiotherapy combined with fruquintinib and tirelizumab in pMMR CRC mice. CT26 cellular hormonal tumor corresponding to pMMR CRC. A mouse model with subcutaneous transplanted tumors is established, divided into control, radiotherapy (IR), fruquintinib + tirelizumab (F+T), and radiotherapy + fruquintinib + tirelizumab (IR+F+T) groups. Immunofluorescence (IF) experiments were conducted to investigate the number and function of tumor vessels. Immunohistochemistry (IHC) and flow cytometry (FC) were utilized to examine immune cell infiltration and alterations within the TIME. Compared to the control group, tumor growth was significantly inhibited after treatment. When compared to IR and F+T, IR+F+T demonstrated a remarkable suppression of tumor growth. The quantitative analysis results showed a significant decrease in Ki-67 positive cells in the target tumors with IR+F+T compared to IR and F+T. The TUNEL results indicated that treatment promoted tumor cell apoptosis, and the effect was further enhanced with triple combinational therapy. Both F+T and IR+F+T repressed CD31 expression and improved the ratio of α-SMA+/CD31+ in tumor tissue, yet there was no significant difference between the two groups. The immunohistochemistry and flow cytometry results demonstrated that triple combinational therapy increased tumor-infiltrating CD8+ T cells and significantly elevated the proportions of CD8, CD69, and CD86. Both F+T and IR+F+T boosted PD-L1 expression, with no significant difference between them. Combined irradiation on top of fruquintinib and tirelizumab treatment enhanced the efficiency in CT26 murine CRC syngeneic tumor model. There was no significant effect of radiotherapy on the anti-angiogenesis of fruquintinib and the promotion of normal vascular function. Irradiation promoted CD8+ T cell and dendritic cell infiltration and activation. Mingsheng Zhang, Qingqing Yu, Huiying Hou, Xiaoting Su, Qin Huang, Hong Qiu, Le Huang, Liang Zhuang, Qiang Fu, Yanmei Zou, Li Sun, Liu Huang, Shunfang Liu, Fei Liu, Xianglin Yuan. Efficacy and mechanism of radiotherapy combined with fruquintinib and tirelizumab in mCRC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1828.

IntroductionInherited retinal diseases (IRDs) affect ∼4.5 million people worldwide. Elusive pathogenic variants in over 280 genes are associated with one or more clinical forms of IRDs. It is necessary to understand the complex interaction among retinal cell types and pathogenic genes by constructing a regulatory network. In this study, we attempt to establish a panoramic expression view of the cooperative work in retinal cells to understand the clinical manifestations and pathogenic bases underlying IRDs.MethodsSingle-cell RNA sequencing (scRNA-seq) data on the retinas from 35 retina samples of 3 species (human, mouse, and zebrafish) including 259,087 cells were adopted to perform a comparative analysis across species. Bioinformatic tools were used to conduct weighted gene co-expression network analysis (WGCNA), single-cell regulatory network analysis, cell–cell communication analysis, and trajectory inference analysis.ResultsThe cross-species comparison revealed shared or species-specific gene expression patterns at single-cell resolution, such as the stathmin family genes, which were highly expressed specifically in zebrafish Müller glias (MGs). Thirteen gene modules were identified, of which nine were associated with retinal cell types, and Gene Ontology (GO) enrichment of module genes was consistent with cell-specific highly expressed genes. Many IRD genes were identified as hub genes and cell-specific regulons. Most IRDs, especially the retinitis pigmentosa (RP) genes, were enriched in rod-specific regulons. Integrated expression and transcription regulatory network genes, such as congenital stationary night blindness (CSNB) genes GRK1, PDE6B, and TRPM1, showed cell-specific expression and transcription characteristics in either rods or bipolar cells (BCs). IRD genes showed evolutionary conservation (GNAT2, PDE6G, and SAG) and divergence (GNAT2, MT-ND4, and PDE6A) along the trajectory of photoreceptors (PRs) among species. In particular, the Leber congenital amaurosis (LCA) gene OTX2 showed high expression at the beginning of the trajectory of both PRs and BCs.ConclusionWe identified molecular pathways and cell types closely connected with IRDs, bridging the gap between gene expression, genetics, and pathogenesis. The IRD genes enriched in cell-specific modules and regulons suggest that these diseases share common etiological bases. Overall, mining of interspecies transcriptome data reveals conserved transcriptomic features of retinas across species and promising applications in both normal retina anatomy and retina pathology.

Alternative splicing of ddx4 (DEAD-box helicase 4), a key germline marker gene, has been reported to generate sex-specific transcripts in zebrafish gonads. The biological functions and regulatory mechanisms of the ddx4 ovary-specific transcript (ddx4-L) during oogenesis remain unclear. In this study, we found that ddx4-L mutants, in which ddx4-L was specifically deleted, had enlarged ovaries but laid fewer eggs, along with having a lower fertilization rate compared to WT controls. RNA-seq analysis was performed to detect the changes in gene expression between WT and ddx4-L mutant ovaries. A total of 524 upregulated and 610 downregulated DEGs were identified. GO and GSEA enrichment analyses showed that genes involved in fertilization and reproduction biological processes were significantly downregulated. More specifically, we observed a remarkable reduction in Sycp1, a core component of synaptonemal complex, in ddx4-L mutant ovaries at both the mRNA and protein levels. In addition, the expressions of transposon elements, as well as the events of alternative splicing, alternative polyadenylation, and RNA editing, were analyzed based on the RNA-seq data. We found that the deletion of ddx4-L resulted in derepression of DNA transposons in zebrafish ovaries, possibly causing genome instability. In conclusion, our work demonstrates that the ovary-specific ddx4 transcript plays important roles in oocyte meiosis and DNA transposon repression, which extends our understanding of the biological functions and regulatory mechanisms of sex-specific alternative splicing in zebrafish oogenesis and reproduction.

Prenatal lethality associated with mouse knockout of Mettl16, a recently identified RNA N6-methyladenosine (m(6)A) methyltransferase, has hampered characterization of the essential role of METTL16-mediated RNA m(6)A modification in early embryonic development. Here, using cross-species single-cell RNA sequencing analysis, we found that during early embryonic development, METTL16 is more highly expressed in vertebrate hematopoietic stem and progenitor cells (HSPCs) than other methyltransferases. In Mettl16-deficient zebrafish, proliferation capacity of embryonic HSPCs is compromised due to G1/S cell cycle arrest, an effect whose rescue requires Mettl16 with intact methyltransferase activity. We further identify the cell-cycle transcription factor mybl2b as a directly regulated by Mettl16-mediated m(6)A modification. Mettl16 deficiency resulted in the destabilization of mybl2b mRNA, likely due to lost binding by the m(6)A reader Igf2bp1 in vivo. Moreover, we found that the METTL16-m(6)A-MYBL2-IGF2BP1 axis controlling G1/S progression is conserved in humans. Collectively, our findings elucidate the critical function of METTL16-mediated m(6)A modification in HSPC cell cycle progression during early embryonic development.

Retinal markers with high quality and specificity are important for the observation of pathologic changes of retinal cells during retinal development, degeneration, and regeneration. The zpr-3 antibody is widely used to label rods in zebrafish, but the exact antigen is still unknown. In this study, we provided evidence to demonstrate that the antigen gene of zpr-3 is rho, which encodes the rod opsin, and the exact epitope of zpr-3 is the 320-354 region of Rho protein. More importantly, our immunofluorescence assays indicated that zpr-3 labels both the outer segments of rods and green cones on zebrafish retinal sections, probably due to the cross-reaction with the green-cone opsin. Our work is valuable for the scientific community to interpret the experimental data involving the zpr-3 antibody.

Background:Several studies have analyzed the relationship between body mass index (BMI) and the prognosis of breast cancer (BC). However, whether their relationship is linear or curvilinear remains unclear. This cohort study examined the specific relationship between BMI and BC outcomes.Methods:This retrospective cohort study included 1049 BC patients from March 7, 2013 through December 31, 2019 in a hospital. Kaplan-Meier curves, multivariate Cox proportional models, and restricted cubic spline (RCS) was used to analysis the relationship between BMI and overall survival (OS) and breast cancer-specific survival (BCSS) was analyzed.Results:During a median of 4.87 (IQR:3.26-6.84) years of follow-up period, 71 patients (6.77%) died, of which 50 (70.42%) were attributed to BC. RCS analysis revealed a U- shaped relationship between BMI levels and OS and BCSS after adjusting for other variables. The turning points of the U-shaped curves were 23 kg/m2. On the left side of the turning point, the risk of OS (HR, 0.83; 95% CI, 0.70, 0.98) and BCSS (HR, 0.80; 95% CI, 0.65, 0.98) were adversely correlated with BMI. In contrast, to the right of the turning point, the risk of OS (HR, 1.22; 95% CI, 1.10, 1.37) and BCSS (HR, 1.28; 95% CI, 1.13, 1.46) was positively related to BMI. Kaplan-Meier curves and multivariate Cox regression analyses shown consistent results with RCS analyses.Conclusion:BMI was an independent prognostic factor for BC, and had a U-shaped relationship with OS and BCSS. Interventions should be designed to improve patient outcomes based on BMI.

INTRODUCTION:Global warming is increasing interest in how aquatic animals can adjust their physiological performance and cope with temperature changes. Therefore, understanding the behavioral changes and molecular underpinnings in fish under warming is crucial for both the individual and groups survival. This could provide experimental evidence and resource for evaluating the impact of global warming. OBJECTIVE:Three genetic families of common carp (Cyprinus carpio) were generated. These juveniles were constructed short-term (4 days) and long-term (30 days) warming groups to investigate the effects of warming on behavioral responses and to elucidate the potential underlying mechanisms of warming-driven behavior. METHODS:Behavioral tests were used to explore the effects of short- and long-term exposure to warming on the swimming behavior of C. carpio. Brain transcriptome combined with measurement of nervous system activity was used to further investigated the comprehensive neuromolecular mechanisms under warming. RESULTS:Long-term warming groups had a more significant impact on the decline of swimming behavior in juvenile C. carpio. Furthermore, brain comparative transcriptomic analysis combined with measurement of nervous system activity revealed that genes involved in cytoskeletal organization, mitochondrial regulation, and energy metabolism are major regulators of behavior in the juvenile under warming. Importantly, especially in the long-term warming groups, enrichment analysis of associated gene expression suggested functional alterations of synaptic transmission and signal transduction leading to swimming function impairment in the central nervous system, as revealed by behavioral tests. CONCLUSIONS:Our study provides evidence of the neurogenomic mechanism underlying the decreased swimming activity in juvenile C. carpio under warming. These findings have important implications for understanding the impacts of climate change on aquatic ecosystems and the organisms that inhabit them.

Mitogen-activated protein kinase (MPK) cascades play vital roles in plant innate immunity, growth, and development. Here, we report that the rice (Oryza sativa) transcription factor gene OsWRKY31 is a key component in a MPK signaling pathway involved in plant disease resistance in rice. We found that the activation of OsMKK10-2 enhances resistance against the rice blast pathogen Magnaporthe oryzae and suppresses growth through an increase in jasmonic acid and salicylic acid accumulation and a decrease of indole-3-acetic acid levels. Knockout of OsWRKY31 compromises the defense responses mediated by OsMKK10-2. OsMKK10-2 and OsWRKY31 physically interact, and OsWRKY31 is phosphorylated by OsMPK3, OsMPK4, and OsMPK6. Phosphomimetic OsWRKY31 has elevated DNA-binding activity and confers enhanced resistance to M. oryzae. In addition, OsWRKY31 stability is regulated by phosphorylation and ubiquitination via RING-finger E3 ubiquitin ligases interacting with WRKY 1 (OsREIW1). Taken together, our findings indicate that modification of OsWRKY31 by phosphorylation and ubiquitination functions in the OsMKK10-2-mediated defense signaling pathway.

Despite the cellular heterogeneity observed in single-cell studies of medulloblastoma, limited knowledge exists regarding the genetic features and spatiotemporal states of specific cell populations following tumor relapse and dissemination, leading to a lack of effective treatment strategies. In this study, we conducted single-cell transcriptomic, chromatin accessibility, and spatial transcriptomic analyses on 15 primary and recurrent Group 3 medulloblastomas, along with two PDX models (including a lung metastasis model). Subsequently, we deconvoluted our findings with our RNA-seq cohort to further validate cell cluster-specific gene expression associated with relapse and its prognostic significance. We analyzed malignant cells, including cancer stem cells (MB-CSCs), proliferating cells, and differentiated cell populations, and examined gene expression features closely associated with clinical prognosis. To elucidate the origin and maturation process of these malignant cells accurately, we compared tumor cell profiles with early human cerebellar cell-types. Furthermore, we described the spatiotemporal transitions from MB-CSCs to terminal states, with a particular emphasis on the extensive spatial heterogeneity within the tumor during the processes of primary, local relapse, and dissemination. Through computational non-negative matrix factorization (cNMF) analysis, we identified a group of tumor cells in recurrent Group 3 medulloblastoma with high cellular plasticity (HCP), demonstrating strong stemness and proliferative capabilities, as well as the ability to differentiate into multiple functional tumor cell subgroups. This cell population exhibited elevated expression of PTPRZ1, which phosphorylates downstream molecules to promote tumor progression. Through single-cell transcriptomic and chromatin accessibility analyses, we validated the expression of PTPRZ1 in human samples, PDX samples, and transgenic mouse samples. In vitro and in vivo inhibition of PTPRZ1 function using small molecule inhibitors effectively suppressed tumor proliferation and metastatic activity. By employing single-cell spatiotemporal genomics, we elucidated the evolutionary states and subclonal events of primary and recurrent medulloblastoma, and identified PTPRZ1 as a target for effectively inhibiting tumor progression.

The swamp eel (Monopterus albus) is a commonly cultured freshwater fish. Selection and cultivation of fecund broodstock is crucial for efficient artificial reproduction in this species. However, there are currently no guidelines for selecting high-quality males in M. albus. This study quantitatively investigated the sperm motility of male swamp eels during the breeding season by using computer-assisted sperm analysis system. Variability in the proportion and number of motile sperm was observed between individual males. The testes were grouped into high-quality and low-quality according to sperm quality. Morphometric parameters of the two groups were analyzed. We found that high-quality testes were longer (150 +/- 27 mm verses 127 +/- 19 mm) than low-quality testes. Besides, the proportion of testes with a dark appearance was slightly higher (58.3% verses 48.1%) in the high-quality group than in the low-quality group. Metabolomic analysis was conducted to compare the metabolite profiles of high-quality and low-quality testes. Glycerophospholipids, organic acids, glycerolipids, amino acids, sphingolipids, and nucleotides were the most abundant metabolites in the testes of swamp eels. Moreover, 135 differential metabolites were identified. Several nucleotide derivatives, including 5'-deoxy-adenosine, 2'-deoxyadenosine, and ribosyl adenosine, were the most significantly enriched metabolites in high-quality testes. These metabolites are potential indicators for the testis quality in M. albus. Altogether, our work provides systematic and quantitative data about the physiological and biochemical characteristics of M. albus testis, which provides an evidential basis for the selection of high-fecundity parents to improve artificial repro-duction efficiency of M. albus.

Hypoxia is typically the leading cause of radiotherapy (RT) resistance in solid tumors, and glutathione (GSH) overexpression in tumor cells is a potent antioxidant mechanism that protects tumor cells from radiation damage. Herein, we developed a sorafenib (SFN) loaded-PLGA hydrogel system (SPH) in combination with microwave (MW) hyperthermia for RT sensitization. SPH with stable properties was produced by combining SFN and PLGA in a specific ratio and encapsulating the mixture in agarose hydrogel. Intratumoral injection of SPH to mice combined with MW hyperthermia can not only directly cause thermal damage to tumor cells, but also increase blood oxygen delivery to the tumor site, thus overcoming the problem of intratumoral hypoxia and achieving "first layer" RT sensitization. Moreover, high temperatures can cause the hydrogel to disintegrate and release SFN. Not only can SFN inhibit tumor growth, but it can also achieve the "second layer" of RT sensitization by inhibiting glutathione (GSH) synthesis in cells and increasing reactive oxygen species (ROS) production. Experiments, both in vitro and in vivo, have indicated that SPH and MW hyperthermia can achieve a double RT sensitization effect and a significant tumor inhibition effect. In conclusion, combining our SPH nanosystem and thermoradiotherapy is a promising anti-tumor treatment.