ABSTRACT Maternal obesity is a risk factor for both maternal and fetal health, and resveratrol (RES) has shown protective effect against adverse pregnancy outcomes in our previous studies. Here, we found that high‐fat diet (HFD) decreased the litter weight (p < 0.05), fetal average weight (p < 0.01), and placenta efficiency (p < 0.01), with increased maternal body weight (p < 0.0001), placental and abdominal lipid accumulation, which were reversed by a supplementation of RES. HFD decreased the angiogenesis in placenta by suppressing the phosphorylation of phosphoinositide 3‐kinase (PI3K), protein kinase B (PKB/AKT), and mammalian target of rapamycin (mTOR), whereas RES reversed the expression of phosphorylation phosphoinositide 3‐kinase (p‐PI3K), phosphorylation protein kinase B (p‐PKB/p‐AKT), and phosphorylation mammalian target of rapamycin (p‐mTOR) (p < 0.05). In vitro cell model confirmed that PI3K inhibitor suppressed the tube formation in embryonic trophoblast cells, and RES can activate PI3K by directly binding to the kinase. These findings demonstrated that maternal obesity impairs placenta angiogenesis, which can be rescued by RES through PI3K phosphorylation.

Betaine acts as a methyl donor and has been proposed as a potential substitute for methionine (Met) to reduce feed costs. This study aimed to investigate the optimal replacement value of betaine for dietary Met in broilers aged 22 to 42 days. A total of 1,440 male broilers were randomly divided into 6 treatments (6 replicates/pen). The positive control (PC) received a standard diet (0.53% Met), while the negative control (NC) received a low-Met diet (0.33% Met) with 2000 mg/kg betaine. Four additional groups received the NC diet supplemented with 0.05%, 0.10%, 0.15%, or 0.20% Met. Data were analyzed using one-way ANOVA and three regression models. Results indicated that Met deficiency significantly impaired growth performance: final body weight (BW) decreased from 1696 g (PC) to 1604 g (NC) (P < 0.001), while the feed-to-gain ratio (F/G) increased from 1.87 to 2.01. No significant differences were observed in slaughter performance (P > 0.05). Regarding metabolism, the NC group exhibited significantly elevated serum homocysteine (19.42 vs. 7.94 μmol/L) and hepatic ALT activity (11.77 vs. 4.35 U/g prot), alongside reduced hepatic BHMT activity (17.47 vs. 103.14 ng/mg prot, P < 0.001). Furthermore, Met deficiency downregulated the mRNA expression of genes related to one-carbon metabolism and intestinal tight junctions (relative expression ∼0.20 vs. 1.00 in PC, P < 0.05). Supplementation with 0.10% Met restored these parameters to levels statistically equivalent to the PC group. One-slope straight broken line and two-slope straight broken line regression analysis of the F/G ratio (R2 = 0.8598) determined the optimal replacement dose to be 0.1474% Met, corresponding to a 27.81% substitution efficiency.

Xuefeng Black-bone chicken is a precious poultry variety for its characteristic traits, high-economic value and medicinal diet value. However, the genomic information for the Xuefeng Black-bone chicken was still lacking, which have hindered the identification of genetic variants underlying its distinctive traits and impeded the implementation of genomic selection strategies in breeding programs. Therefore, we applied PacBio SMRT (Single Molecule Real-Time) sequencing, Illumina sequencing, Hi-C and RNA-seq to establish a high-contiguity reference genome for Xuefeng Black-bone chicken, achieving chromosome-level. The de novo assembly yielded a 1.13 Gb genome, exhibiting contiguity metrics of 21.76 Mb (contig N50) and 83.79 Mb (scaffold N50), and comprised 40 pseudochromosomes. Merqury demonstrated a QV (Consensus quality value) score of 43.1876 and 96.7

The experiment aimed to investigate the effects of partially replacing soybean meal with deep enzymatically hydrolyzed soybean meal (ESBM) on the growth performance, immune function, and cecal microbiota of weaned piglets. A total of 192 healthy crossbred weaned piglets at 28 days of age with an initial body weight of (7.01±0.08) kg were randomly divided into four groups, with six replicates per group and eight piglets per replicate. The control group (group CON) was fed a basal diet, while the experimental groups were fed the basal diet in which part of the soybean meal extruded soybeans was replaced by 5% (group ESBM1), 10% (group ESBM2), and 15% (group ESBM3) ESBM on an isoenergetic and isonitrogenous basis, respectively. The test period was 42 days. The results showed that compared with the group CON, the average daily gain (ADG) and final body weight in all experimental groups were significantly increased (P<0.05). The average daily feed intake (ADFI) in the groups ESBM2 and ESBM3 was significantly increased (P<0.05), while the feed-to-gain ratio (F/G) in the groups ESBM1 and ESBM2 was significantly decreased (P<0.05), and the diarrhea index in the group ESBM1 was significantly reduced (P<0.05). Compared with the group CON, the serum immunoglobulin M (IgM) content in the groups ESBM2 and ESBM3 was significantly decreased (P<0.05), the immunoglobulin E (IgE) content in the group ESBM2 was significantly decreased (P<0.05), the tumor necrosis factor-α (TNF-α) content in the group ESBM1 was significantly decreased (P<0.05). In the group ESBM3, the concentrations of interleukin-10 (IL-10) and lipopolysaccharide (LPS) were significantly increased (P<0.05), while the interferon-γ (IFN-γ) content was significantly decreased (P<0.05). Compared with the group CON, the Sobs, Shannon index, and Ace index in all experimental groups were significantly increased (P<0.05), and the Beta-diversity of cecal microbiota was significantly altered (P<0.05), with an increased relative abundance of Bacteroidetes in the cecum. The study shows that ESBM can improve growth performance, reduce diarrhea index, and enhance intestinal health in weaned piglets, while its regulatory effects on immune inflammation and intestinal barrier-related indicators exhibit a dose-dependent pattern, and the optimal replacement level of ESBM for soybean meal in the diet of weaned piglets is 10%.

This study aims to investigate the effect of iron overload on intestinal damage in broilers infected with Eimeria tenella. A total of 240 one-day-old white-feathered broilers (Arbor Acres, AA) were randomly assigned to four groups based on a 2 × 2 factorial design (2 dietary iron levels: 80 or 500 mg/kg; Eimeria tenella challenge: with or without), with six replicates per group, 10 birds per replicates. The dietary treatments were: 1) control group (CON) and attack group (AG): basal diet + 80 mg/kg FeSO4. 2) high iron group (HF) and high iron + attack group (HFA): basal diet + 500 mg/kg FeSO4. On day 18, broilers in the AG and HFA groups were challenged with 8 × 104 sporulated oocysts of Eimeria tenella, while broilers in CON and HF groups received Phosphate Buffered Saline (PBS). Our results indicates that: 1) Compared to the others, HFA had the lowest survival rate and the highest Eimeria fecal oocysts (P < 0.001) and showed lower expression of intestinal tight junction proteins (ZO-1, Claudin-1, and Occludin) in broilers (P < 0.05). 2) The interaction between these factors indicated that iron overload exacerbated the expression of IL-8 (P < 0.001) and IL-1β (P = 0.025) on 4 dpi, IL-1β (P = 0.023) on 8 dpi. 3) Both the Eimeria tenella challenge and the high-iron diet led to a significant increase in serum levels of DAO and LPS, while simultaneously decreasing the levels of IgA, IgG, and IgM (P < 0.05). 4) WGCNA-GSEA analysis revealed that the calcium signaling pathway, Wnt signaling pathway, and MAPK signaling pathway were specifically upregulated in HFA, while the TCA-cycle and fatty acid metabolism and degradation pathways were specifically downregulated. 5) Both the high-iron diet and Eimeria tenella challenge significantly altered the α-diversity and β-diversity of cecal microbiota (P < 0.05). Moreover, they significantly enriched microbial functions related to vancomycin resistance (ko01502), peptidoglycan biosynthesis (ko00550), and galactose metabolism (ko00052). In conclusion, this study indicates that iron overload exacerbates intestinal damage in broilers challenged by Eimeria tenella, potentially through its influence on the calcium signaling pathway in cecal tissue and altering both the structure and function of the cecal microbiota. An appropriate reduction in iron levels in poultry diets may contribute to enhanced coccidiosis control within the poultry industry.

The molecular characterization of chicken cells provides insight into avian physiology and vertebrate evolution. Here we present a single-cell RNA-sequencing (scRNA-seq) atlas of the chicken, encompassing 1.57 million cells across 157 cell types from 36 tissues from 16 chickens, as well as spatial transcriptomes from 1 chicken embryo. We further acquire scRNA-seq data from 3 turtles (230 thousand cells, 14 tissues) and 2 ducks (22 thousand cells, 3 tissues), and integrated our data with publicly available human scRNA-seq data (960 thousand cells, 32 tissues) to explore their evolutionary conservation, particularly of immune cells. In chicken, follicular dendritic cells, unlike in mammals, exhibit myeloid rather than stromal origins and help promote B cell proliferation and migration in the bursa of Fabricius. γδ T cell subsets vary across species, reflecting the evolution of pathogen recognition and signaling mechanisms among amniotes. Our transcriptomics data thus provide a resource to study chicken cell biology and amniote evolution, highlighting immune cell similarities and distinctions arising from evolutionary divergence.

The domestic chicken (Gallus gallus) is a critical model for immunology and a major agricultural species, yet a comprehensive single-cell census of its immune architecture is lacking. Here, we constructed a high-resolution single-cell transcriptome atlas of three primary immune organs (thymus, bursa of Fabricius, and spleen) from chickens. Our analysis identified 24 distinct immune and stromal cell types, mapping their organ-specific distributions: B cells were predominantly localized to the bursa of Fabricius, while T cell subsets were enriched in the thymus. We delineated developmental trajectories and identified key transcription factors governing lineage commitment, including TCF12 for CD4⁺CD8⁺ T cells and SOX13 for T helper 2 cells. Further analysis revealed potential sexually dimorphic immune maturation, where B cells exhibited sex-biased gene expression primarily in early differentiation stages, whereas T cell differences were most pronounced during proliferation. Cross-species comparison demonstrated remarkable conservation of core immunoregulatory circuits, with transcription factors like MEF2C, LEF1, and GATA3 playing conserved roles in avian and mammalian immunity. This resource provides a foundational framework for understanding avian immune defense, advancing poultry disease-resistant breeding, and offers evolutionary insights into vertebrate immunology.

The present study aimed to evaluate the effects of zinc amino acid complexes on growth performance, tissue zinc concentration, and muscle development in broilers. A total of 504 day-old male arbor acres broilers were randomly divided into seven treatments (fed with a basal diet or a basal diet supplemented with 120 mg kg −1 Zn as ZnSO 4 , 30, 60, 90 or 120 mg kg −1 Zn as ZnN, or 30 mg kg −1 Zn as ZnA separately). Each group had six replicates, with 12 birds per replicate. The results showed that the addition of 60 mg kg −1 ZnN significantly increased ( P < 0.05) the average daily gain (ADG) and breast muscle percentage of broilers. Zinc concentration of ZnN and ZnA added groups were higher than ( P < 0.05) that in the Zn sulfate group under the same addition dose. Except for the 30 mg kg −1 ZnN group, the muscle fiber diameter and cross-sectional area (CSA) were significantly increased ( P < 0.05) in the ZnN addition groups. Compared with the basal diet group, adding ZnN significantly increased ( P < 0.05) the expression of MTOR, MYOD, and MYOG at day 21 and decreased ( P < 0.05) the expression of Atrogin-1. The expression levels of AKT, MTOR, P70S6K, and MYOD were increased at day 42, while the expression levels of MuRF1 and Atrogin-1 were decreased. Adhesion, backbone regulation of actin, MAPK, mTOR, and AMPK were significantly enriched as indicated by KEGG pathway enrichment analysis. In conclusion, zinc amino acid complexes could improve growth performance, tissue zinc concentration, and regulate breast muscle development.

Background Embryonic muscle development is a highly dynamic and complex process, coordinated by numerous genes and transcriptional regulators such as small RNAs. Results Here, we profiled transcriptomic dynamics in breast muscle from Arbor Acres (AA) broilers and TaoYuan (TY) chickens at three embryonic time points (E9, E13, and E18). While developmentally regulated genes enriched similar biological pathways in both breeds, the architecture of microRNA (miRNA)-mRNA interaction networks was distinct. Integrative analysis combining weighted gene co-expression network analysis, differential expression analysis, and time-series clustering identified 161 candidate genes, including a subset of 39 showing progressively decreasing expression. Differential expression analysis of miRNA revealed that gga-miR-1744-3p was uniquely up regulated at E13 and E18 in TY chickens. By integrating predictions from TargetScan and miRDB, we further identified two core genes (USP8 and ZBTB38) from the set of 39 candidate genes, which are predicted targets of gga-miR-1744-3p. This gene was differentially expressed in the heart, breast muscle, and adipose tissue, and exhibited significantly higher expression in TY chickens. Functional assays confirmed that gga-miR-1744-3p promotes proliferation of chicken primary myoblasts. Conclusions This study provides a comprehensive understanding of the development patterns and molecular mechanisms of breast muscles in broilers during embryogenesis.

Sexual dimorphism is a defining vertebrate feature, yet its sex-specific molecular architecture remains poorly understood. Here we established a sex-balanced, uniformly reared chicken cohort to map this landscape, integrating individual whole-genome sequencing with 7,969 bulk and 779,380 single nucleus transcriptomes across 32 tissues from 280 birds. We identified 495,098 independent expression quantitative trait loci for 20,194 genes, including 10,937 loci modulated by cell-type composition. Notably, 340 genes were regulated by 449 loci in a sex-dependent manner, significantly enrichment in endocrine tissues like adipose and the adrenal gland. Furthermore, we fine-mapped 1,219 structural variants, demonstrating their unique roles to tissue- and sex-specific expression beyond SNPs. Ultimately, we showed the utility of these regulatory effects in elucidating the molecular basis of metabolism and complex traits in both chickens and humans. This comprehensive atlas of regulatory effects provides profound insights into the genomic and molecular basis of sexual dimorphism in vertebrates.

To better understand the regulatory mechanisms of breast muscle growth in small-sized local breeds, this study aimed to investigate metabolic and transcriptional networks during the initiation of sexual maturation in Huanglang chickens. We hypothesized that sex-specific metabolic and gene expression patterns regulate muscle growth and fat deposition in these chickens. To test this hypothesis, multi-omics approaches were used to analyze chickens at 80 and 120 days post-hatch (dph). Both male and female chickens showed a significant increase in intramuscular fat (IMF) at 120 dph, but with sex-specific changes: females exhibited a significantly higher liver index, while males had a significantly greater breast muscle index. We identified 2627 differentially expressed genes (DEGs) in males and 2991 in females, along with 473 and 232 differentially abundant metabolites (DAMs), respectively. The sex-shared ABC transporter pathway supports muscle growth via substrate transport, while the Steroid biosynthesis pathway is female-specific, and the Glycerophospholipid metabolism pathway is male-specific. These results demonstrate that sex-specific regulatory networks shape muscle growth and fat deposition during early sexual maturation, and they provide potential molecular targets for improving intramuscular fat content and meat quality in local chicken breeding programs.

IntroductionConventional fasting molting can restore laying performance but imposes substantial physiological stress and welfare concerns. High-zinc diets (2% ZnO) have been proposed as a less stressful alternative, but their mechanisms remain unclear.MethodsThis study compared fasting-induced molting with zinc oxide supplementation in 384 Lohmann Pink hens (65 wk). Serum, cecal contents, and ovaries were sampled across six stages from pre-molt to post-molt recovery.ResultsZnO accelerated cessation of lay (5.75 vs. 8.87 d) and earlier recovery to 50% production (15.25 vs. 16.62 d) with lower weight loss (25% vs. 30%). Multi-omics revealed that high-zinc feeding enriched beneficial microbes (e.g., Coprenecus pullicola, Fournierella spp.) involved in amino acid and cofactor biosynthesis, consistent with activated glycine/serine/lysine metabolism. ZnO also reduced inflammatory and barrier-injury signals (IL-1β, LPS, DAO) and promoted earlier recovery of reproductive hormones (higher IGF-1, earlier increases in E2 and LH). Both treatments improved post-molt egg quality; zinc better preserved albumen height and Haugh unit, while fasting increased shell thickness and yolk color.Discussion/conclusionThese findings support that a high-zinc-associated microbial-metabolic profile mitigates stress, accelerates ovarian reset, and shortens the time to restore production.

Ex vivo expansion of human hematopoietic stem cells (HSCs) holds promise for overcoming their limited availability, a major barrier to broader clinical application. Although recent advances in culture systems can increase HSC numbers, these conditions frequently impair self-renewal and induce myeloid bias, and the underlying molecular mechanisms remain poorly understood. Here, we performed single-cell multiome sequencing (scMultiome-seq) on human umbilical cord blood-derived CD34⁺ hematopoietic stem and progenitor cells to co-profile transcriptional and epigenetic adaptations within the same cells during ex vivo culture. Our analyses revealed reduced transcriptional and epigenetic HSC signatures, accompanied by markedly increased activity of myeloid-associated transcription factor motifs, providing molecular insight into the functional decline and myeloid bias of cultured HSCs. We further observed substantial functional heterogeneity among phenotypically defined HSCs following culture. To address these limitations, we established a niche-mimetic culture system that integrates intrinsic and extrinsic bone marrow regulatory cues, including pharmacologic inhibition of the m6A reader YTHDF2 using the small molecule Y13-27, a three-dimensional microenvironment, and N-cadherin-mediated adhesion. This condition (3D-NcadP-Y) robustly preserved long-term repopulating capacity. When combined with the self-renewal agonist UM729, the resulting platform (3D-NcadP-Y-UM) uniquely enabled the expansion of serially transplantable long-term HSCs with balanced multilineage potential. scMultiome-seq and cellular analyses demonstrated that this condition preserves transcriptional and epigenetic long-term HSC signatures, maintains multilineage-associated transcription factor motifs, and limits excessive cell-cycle activation. Together, these findings elucidate molecular mechanisms underlying culture-induced HSC dysfunction and establish a niche-mimetic strategy for expanding functional human long-term HSCs while preserving key features of stemness.

The rice seed storability gene Seed storability 7 (SS7) encodes a cytosol localized sucrose synthase (SUS) domain protein. SS7 positively regulates seed storability by enhancing the activity of peroxidase and nitrate reductase (NR) in seeds, resulting in the reduction of ROS and NO accumulation during seed storage. Long-term storage of crop seeds is essential for conserving germplasm resources, ensuring food security, and supporting sustainable agriculture. In this study, we characterized the function of Seed storability 7 (SS7), the causal gene for the QTL qSS7-1 responsible for rice seed storability identified by genome-wide association studies (GWAS). SS7 encodes a cytosol localized sucrose synthase (SUS) domain protein. Overexpression of SS7 significantly enhanced seed storability under both artificial and natural aging conditions. While knockout of SS7 has no significant effects in seed storability, but showed a phenotype of enhanced elongation of rice root. Transcriptome analysis revealed that differentially expressed genes (DEGs) involving in the hydrogen peroxide (H₂O₂) catabolic pathway were significantly enriched in SS7 overexpression lines; in contrast, the DEGs in both H₂O₂ catabolism and nitric oxide (NO) biosynthesis pathways were enriched in SS7 knockout lines. Furthermore, ROS accumulated significantly in SS7 knockout seeds, whereas SS7 overexpression in seeds resulted in a significant enhancement of peroxidase activity and a significant reduction in nitrate reductase (NR) activity, compared to SS7 knockout seeds. These results suggested that SS7 enhances seed storability by reducing the accumulation of peroxides, but inhibits root elongation by promoting NO accumulation, of which provides new insights into understanding the molecular mechanisms of rice seed storability.

Boar semen quality is a significant factor affecting reproductive efficiency in modern farms, and resveratrol (RES) has the potential to reduce mouse sperm mortality in our previous studies. Here, we found that dietary supplementation of RES (500 mg/kg) delayed the decline in semen quality by increasing sperm motility and survivability in aging boars (P < 0.01, n = 6) by a 12-wk trial. RES increased the testosterone level and total antioxidant capacity (P < 0.01), while decreasing the cortisol (P < 0.05) and malondialdehyde (MDA, P < 0.01) in serum. Immunofluorescence assay and electron microscope scanning revealed that RES reduced the levels of reactive oxygen species, 8-hydroxy-2′-deoxyguanosine, and MDA (P < 0.01) to alleviate oxidative damage in semen. Analysis of mitochondrial function-related genes indicated that RES improved mitochondrial structural integrity and mitochondrial membrane potential (P < 0.01) by increasing the mRNA expression of PGC-1α, TFB1M, and TFB2M (P < 0.01). These results demonstrated that RES can enhance semen quality and antioxidant capacity potentially through improving sperm mitochondrial function in aging boars.

Abstract Background Liver lipid dysregulation is one of the major factors in the decline of production performance in late-stage laying hens. Silymarin (SIL), a natural flavonolignan extracted from milk thistle, is known for its hepatoprotective and lipid-lowering properties in humans. This study evaluates whether SIL can provide similar benefits to late-stage laying hens. A total of 480 68-week-old Lohmann Pink laying hens were randomly assigned into 5 groups, each group consisting of 6 replicates with 16 hens each. The birds received a basal diet either without silymarin (control) or supplemented with silymarin at concentrations of 250, 500, 750, or 1,000 mg/kg (SIL250, SIL500, SIL750, SIL1000) over a 12-week period. Results The CON group exhibited a significant decline in laying rates from weeks 9 to 12 compared to the initial 4 weeks (P = 0.042), while SIL supplementation maintained consistent laying rates throughout the study (P > 0.05). Notably, the SIL500 and SIL750 groups showed higher average egg weight than the CON group during weeks 5 to 8 (P = 0.049). The SIL750 group had a significantly higher average daily feed intake across the study period (P < 0.05), and the SIL500 group saw a marked decrease in the feed-to-egg ratio from weeks 5 to 8 (P = 0.003). Furthermore, the SIL500 group demonstrated significant reductions in serum ALT and AST levels (P < 0.05) and a significant decrease in serum triglycerides and total cholesterol at week 12 with increasing doses of SIL (P < 0.05). SIL also positively influenced liver enzyme expression (FASN, ACC, Apo-VLDL II, FXR, and CYP7A1; P < 0.05) and altered the cecal microbiota composition, enhancing species linked to secondary bile acid synthesis. Targeted metabolomics identified 9 metabolites predominantly involved in thiamin metabolism that were significantly different in the SIL groups (P < 0.05). Conclusions Our study demonstrated that dietary SIL supplementation could ameliorate egg production rate in late stage laying hens, mechanistically, this effect was via improving hepatic lipid metabolism and cecal microbiota function to achieve. Revealed the potentially of SIL as a feed supplementation to regulate hepatic lipid metabolism dysregulation. Overall, dietary 500 mg/kg SIL had the best effects. Graphical Abstract

Genetic mutation and drift, coupled with natural and human-mediated selection and migration, have produced a wide variety of genotypes and phenotypes in farmed animals. We here introduce the Farm Animal Genotype-Tissue Expression (FarmGTEx) Project, which aims to elucidate the genetic determinants of gene expression across 16 terrestrial and aquatic domestic species under diverse biological and environmental contexts. For each species, we aim to collect multiomics data, particularly genomics and transcriptomics, from 50 tissues of 1,000 healthy adults and 200 additional animals representing a specific context. This Perspective provides an overview of the priorities of FarmGTEx and advocates for coordinated strategies of data analysis and resource-sharing initiatives. FarmGTEx aims to serve as a platform for investigating context-specific regulatory effects, which will deepen our understanding of molecular mechanisms underlying complex phenotypes. The knowledge and insights provided by FarmGTEx will contribute to improving sustainable agriculture-based food systems, comparative biology and eventual human biomedicine.

The experiment aimed to determine the apparent metabolizable energy (AME) and nitrogen-corrected apparent metabolizable energy (AMEn) of 10 different sources of Sorghum bicolor L. Moench for fast-growing yellow-feathered chickens, and construct predictive models for AME and AMEn. A total of 792 one-day-old healthy Xiangjia Huang No.2 male broilers were randomly divided into one basal diet group and 10 Sorghum bicolor L. Moench experimental diet groups (experimental diets some energy components were replaced with 30% Sorghum bicolor L. Moench). The pre-test period was three days, and the formal test period was three days. The experiment was divided into two stages: The first stage (12 to 18 days old), 528 eleven-day-old broiler chickens were selected with six replicates per group and eight chickens per replicate. The second stage (33 to 39 days old), 264 thirty-two-day-old broiler chickens were selected with six replicates per group and four chickens per replicate. The total excretae collection method was used to measure the metabolizable energy values. The results showed that the average content of the main components in 10 Sorghum bicolor L. Moench varieties was: Dry matter (DM) 87.36%, crude protein (CP) 10.10%, crude fat (EE) 3.63%, neutral detergent fiber (NDF) 12.01%, acid detergent fiber (ADF) 3.92%, crude ash (Ash) 1.66%, crude fiber (CF) 2.46%, tannin 0.78%, phytic acid (PA) 0.85%, nitrogen-free extract (NFE) 69.50%, starch 58.74%, and gross energy (GE) 18.66 MJ/kg (dry matter basis). During the 12 to 18 days, the average AME and AMEn values for the 10 Sorghum bicolor L. Moench varieties were 12.61 MJ/kg and 12.53 MJ/kg (dry matter basis), respectively. During the 33 to 39 days, the average AME and AMEn values were 14.66 MJ/kg and 14.59 MJ/kg (on a dry matter basis), respectively. At 18 days of age, the model equations were: AME=2.319+1.102CP-1.781Pro (R2=0.919, P=0.001), AMEn=2.295+1.096 CP-1.751Pro (R2=0.921, P<0.001). At 39 days of age, the model equations were: AME=14.691-0.994tannin+2.929Cys (R2=0.837, P=0.004), AMEn=14.612-1.001tannin+3.004Cys (R2=0.834, P=0.005). The model validation showed accurate predictive values. The study indicates that CP and tannin content are the key factors affecting the metabolizable energy of Sorghum bicolor L. Moench in Xiangjia Huang No.2 male broilers at 18 and 39 days of age, respectively.

Abstract Leukemia stem cells (LSCs) are key drivers of chemoradiotherapy resistance, immune evasion, and frequent relapse of leukemia. Despite achieving initial remission, 30-50% of acute myeloid leukemia (AML) patients eventually relapse, even after allogeneic stem cell transplantation. Relapse samples show a 10- to 90-fold increase in LSC frequency, further implicating LSC's role in leukemia recurrence. The Wnt/β-catenin and PI3K/AKT pathways are frequently dysregulated in many cancers, and are crucial for LSCs' self-renewal and immune escape. β-catenin directly regulates the expression of multiple immune checkpoint genes. Additionally, AKT promote Wnt/β-catenin by phosphorylating and inhibiting GSK-3β to promote β-catenin accumulation, and directly phosphorylating β-catenin at Ser552 and Ser675 to enhance the nuclear transcriptional activity of β-catenin. Thus, hyperactivating the AKT-Wnt/β-catenin signaling plays a critical role in supporting LSCs' self-renewal and survival. The intricate relationship between the AKT-Wnt/β-catenin pathway and the mechanisms of immune escape and cancer recurrence underscores the urgent need for targeted interventions. We previously demonstrated that low-dose doxorubicin inhibits AKT-mediated Ser552 phosphorylation of β-catenin, and reduces LSCs in both T-cell acute lymphoblastic leukemia (T-ALL) mouse models and in a pilot clinical trial in refractory AML patients. However, due to topoisomerase-II binding, doxorubicin's cardiotoxicity limits its clinical utility. Over 20% of pediatric ALL survivors develop grade II cardiotoxicity post-chemoradiotreatment. Peripheral neuropathy occurs in approximately 38% of patients receiving neurotoxic chemotherapies, causing long-term sensory and motor deficits. Hence, there is an urgent need for safer LSC-targeting strategies. To address these challenges, we developed Artificial intelligence-powered, mRNA-delivery of Peptides (ARP), an integrated approach that combines AI-guided peptide designing with lipid-nanoparticle (LNP)-facilitated mRNA delivery for precise intracellular targeting of the AKT-β-catenin interaction. Leveraging AlphaFold-simulated docking in silico, and reporter assays in HEK293-AKT-β-catenin-TOPFlash-luciferase cells in vitro, we validated several highly potent peptide inhibitors, including β-catenin fragment 529-581. Next, we used RFdiffusion to generate 2,000 de novo binders targeting two hydrophobic hotspots within the AKT kinase-pocket. Following a streamlined de novo binder screening pipeline in silico, including competitive binding assay, immunogenicity and toxicity filters, we selected the top 35 candidates. Meanwhile, we created a library of 416 β-catenin fragments centered on the AKT phosphorylation site at Ser552. We then trained a machine-learning model by integrating (1) reporter assay data from these fragments in HEK293-AKT-β-catenin-TOPFlash-dGFP cells and (2) six structural and energetic features obtained from their AlphaFold3 docking conformations. Through this model, we ranked the last 35 candidates and predicted the top-ranked de novo peptide, Dnv, demonstrated its superior binding and stability in silico, potent inhibition in reporter cells, and enhanced stability relative to 529-581 in vitro. ARPs were then tested in SclCreER;Ptenfl/f;Ctnnb1Dex/+3(β-catenin) T-ALL mouse model. Linear or circular RNA encoding β-catenin fragments or Dnv was packaged in LNP and intravenously administered. ARP treatments were well tolerated and significantly reduced LSCs, partially restored hematopoietic stem/progenitor cells without obvious cardiotoxicity or hepatotoxicity. Competitive transplantation assays revealed that unlike control, ARP-treated bone marrow cells didn't drive leukemogenesis. ScRNA-seq of bone marrow cells confirmed a significant decrease in LSCs. Moreover, we found a pronounced shift of CD8⁺ T-cells that β-catenin fragments or Dnv treatment decreased stem-like CD8+ T memory cells, which are induced by elevated Wnt/β-catenin signaling. Effector and effector memory cells expanded, and their expressions of anti-tumor cytotoxic genes were elevated, accompanied by downregulation of exhaustion markers. Together, these results establish ARP as a versatile strategy to target intracellular AKT-Wnt/β-catenin signaling. By simultaneously disrupting LSCs' maintenance and reprogramming anti-tumor immunity, ARP offers a promising avenue for overcoming relapse and enhancing therapeutic responsiveness in leukemia.

Sexual maturity significantly impacts poultry production efficiency, yet data on testicular development and regulatory mechanisms in indigenous breeds remain limited. In this study, we examined Xianghuang chickens, a indigenous early-maturing breed, to investigate the hypothalamus-pituitary-testis (HPT) axis. Hypothalamus, pituitary, and testis tissues were collected at 80 and 120 days post-hatch (dph) for analysis. Results showed that testicular weight and index were significantly higher in 120dph compared to 80dph chickens. Histologically, the testes at 120dph contained multi-layered spermatocytes and mature sperm while the testes at 80dph only had 1-2 layers of spermatocytes. Weighted Gene Co-expression Network Analysis of transcriptome data clustered gene sets into 11 modules including those with testis-specific high expression, 80dph testis-specific high expression, and 120dph testis high expression. Differential expression analysis identified 26, 681, and 11,999 differentially expressed genes (DEGs) in the hypothalamus, pituitary, and testes respectively between the two age groups with 16 DEGs shared among the three tissues. DEGs in all three tissues were enriched in pathways such as cytoskeleton in muscle cells, ECM-receptor interaction, and focal adhesion highlighting the importance of HPT axis tissue remodeling during this period. Hormone-related pathways (GnRH signaling, melanogenesis, and progesterone-mediated oocyte maturation) were enriched with DEGs in the pituitary and testes between 80dph and 120dph . Concurrently the pituitary and testes exhibited distinct lipid metabolic adaptations through enrichment of PPAR, adipocytokine, and insulin signaling pathways. These adaptations provided substrates for sperm membrane formation and testosterone synthesis while supporting spermatogenesis and cellular functions. Collectively these findings clarify the morphological, histological, and molecular regulatory mechanisms underlying testicular development and sexual maturation in Xianghuang chickens and offer a theoretical basis for optimizing poultry breeding strategies.