Yong Liu

The spatiotemporal control of asymmetric positioning of cellular cargoes, such as macromolecules and organelles, during spermatogenesis is essential for producing functional sperm in most animal species. However, the regulatory mechanisms that regulate cargo sorting and transporting remain largely unknown. Here, we demonstrate that the casein-like kinase SPE-55 localized on the surface of a membranous organelle (MO) is required for the SPE-18 localization/puncta formation and facilitates the assembly of major sperm protein (MSP) condensate, named the fibrous body (FB), in spermatocytes during spermatogenesis in Caenorhabditis elegans . Furthermore, we discover that the H subunit of v-ATPase, which is localized on MOs, played a crucial role in maintaining MO morphology and function, promoting FB–MO connections, and thus ensuring efficient MSP sorting during spermatogenesis. Therefore, the interplay between the membraneless condensate FBs mediated by SPE-55 and the membrane-bound MOs regulated by VHA-18 provides a coordinated mechanism for cargo sorting and directional transport during spermatogenesis in C. elegans .

Follicular atresia constitutes a major constraint on sow reproductive performance. Aberrant miRNA expression is well documented to promote follicular atresia. However, precise identification of the functional miRNAs responsible for this process remains a formidable challenge. In this study, we integrated multiple transcriptome datasets and identified miR-149, a miRNA located within the quantitative trait loci (QTL) associated with litter size in sow. Notably, miR-149 was highly expressed in atretic follicles and ovaries of sow with low litter size. Subsequent in vitro culture of ovarian granulosa cells (GCs) combined with miR-149 overexpression assays revealed that miR-149 significantly promotes the expression of cleaved caspase 3 (c-caspase 3), a marker of apoptosis in ovarian GCs. Mechanistic studies revealed that miR-149 functions as small activating RNA (saRNA) to activate the transcription of Caspase 8. This triggers the death receptor-mediated apoptotic, upregulates the expression of Caspase 3, and ultimately accelerates apoptosis in ovarian GCs. Collectively, these results indicate that miR-149 acts as negative regulator of sow reproduction. This study elucidates the miR-149/Caspase 8/Caspase 3 signaling axis as a novel pathway governing sow fertility, which provides molecular targets for improving reproductive performance in sow.

The corpus luteum number in sows is a key reproductive indicator for measuring ovulation rate and reproductive efficiency. Its formation is precisely regulated by a complex gene regulatory network composed of multi-level molecular interactions. To systematically elucidate the molecular basis of this trait, this study comprehensively analyzed genes located within QTL for corpus luteum number. This approach identified a series of key regulatory molecules specifically expressed in the ovary, including transcription factors (TFs), long non-coding RNAs (lncRNAs), and microRNAs (miRNAs). Using bioinformatics methods to predict the target genes of candidate miRNAs, combined with functional enrichment analysis, revealed that these target genes were significantly enriched in multiple core reproductive pathways closely related to cell proliferation, differentiation, and hormone regulation, including the ErbB signaling pathway, PI3K-Akt signaling pathway, and TGF-beta signaling pathway. Based on the above findings, this study ultimately constructed a TF-lncRNA-miRNA-mRNA network, which is associated with QTL for corpus luteum number. Furthermore, key genes were validated via quantitative real-time PCR (qRT-PCR). Significant positive correlations were identified between the transcription factor NEUROG2 and lncRNA LOC102167554, along with its potential target gene ESRP1, as well as between transcription factor SNAI2 and lncRNAs (LOC102167554, LOC102167796) and their potential target genes (FXR, ERBB4). In addition, the functional validation results showed that the interference of LOC102167554 significantly reduced the proliferation ability of sGCs. These key genes represent potential targets for genetic improvement of sow reproduction.

Objectives: Postmenopausal osteoporosis is the most common form of osteoporosis in clinical practice, affecting millions of postmenopausal women worldwide. Postmenopausal osteoporosis demands safe and effective therapies. This study aimed to evaluate the potential of hederagenin (Hed) for treating osteoporosis and to elucidate its underlying mechanisms of action. Methods: The anti-osteoporotic potential of Hed was assessed by investigating its effects on ovariectomy (OVX)-induced bone loss in mice and on receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast differentiation in RAW264.7 cells. Network pharmacology analysis and molecular docking were employed to identify key targets, which were subsequently validated experimentally. Results: In vitro, Hed suppressed osteoclastogenesis by inhibiting the formation of osteoclasts and F-actin rings and by down-regulating osteoclast-specific genes (Atp6v0d2 and Acp5). In vivo, Hed significantly ameliorated OVX-induced bone loss, restoring trabecular bone volume fraction (BV/TV) and trabecular number (Tb.N), while reducing trabecular separation (Tb.Sp). Network pharmacology analysis identified 142 overlapping targets linking Hed to osteoporosis, including tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and IL-1β, with enrichment in innate immune signaling and osteoclast differentiation. Molecular docking analysis indicated strong binding affinities between Hed and targets such as TNF-α, IL-6, and IL-1β. Experimentally, Hed was found to decrease RANKL, elevate osteoprotegerin (OPG), and suppress intestinal mRNA levels of pro-inflammatory cytokines such as IL-1β, IL-6, IL-17A, and TNF-α. Conclusion: Hed exerts significant anti-osteoporotic effects in OVX-induced osteoporosis through a dual mechanism involving the suppression of both osteoclastogenesis and innate immune signaling pathways. These findings highlighted Hed’s novel role in modulating immune-bone crosstalk, offering a promising strategy for treating osteolytic diseases without estrogenic side effects.

The quality of muscle is a significant economic trait in livestock, the development of which is contingent on the coordinated regulation of both coding and non-coding genes. The Anhui White Goat is a breed that is native to central China. It is characterised by its high levels of adaptability and tolerance, which enable it to thrive under extensive management conditions. Furthermore, it is renowned for producing meat that is particularly flavourful. However, in comparison with the growth rate and meat yield of introduced breeds such as the Boer goat, the local breed exhibits a lower level of productivity. In the present study, we conducted a comprehensive analysis of the transcriptomes of longissimus dorsi muscle samples from Anhui White Goats and Boer Goats. We identified 490 key candidate microRNAs associated with muscle growth and development, among which 19 were differentially expressed (|log₂FC| ≥ 1.5, P < 0.05), including 7 up-regulated and 12 down-regulated microRNAs. Further prediction of the target genes of differentially expressed miRNAs revealed that these miRNAs target genes such as BDNF, MAPT, MYB, STC1, and SMAD7. These target genes are primarily involved in signaling pathways including MAPK, GABAergic synapse, HIF-1, Toll-like receptor, PI3K/Akt, and cAMP. Subsequent refinement revealed five key candidate microRNAs potentially involved in this process: chi-miR-10b-5p, chi-miR-122, chi-miR-novel-7_9719, chi-miR-143-3p, and chi-miR-424-3p. These findings provide a theoretical foundation for further elucidating the molecular mechanisms underlying muscle growth and development in Anhui White Goats.

Black carbon has attracted significant attention because of its severe health hazards. Carbon black (CB), a commercially available standardized particulate material, is widely used as a surrogate model in toxicological studies. The voltage-gated proton channel Hv1, encoded by the Hvcn1 gene, is specifically expressed in lung epithelial cells and regulates the generation of reactive oxygen species. However, the role of Hv1 in lung homeostasis remains unclear. In this study, we constructed an Hv1 knockout (KO) mouse model via CRISPR/Cas9 technology to investigate the impact of channel deficiency on lung injury induced by exposure to CB particles. Our findings revealed that Hv1 deficiency significantly exacerbated lung injury caused by CB particles compared with that in wild-type (WT) mice. Specifically, Hv1 knockout mice presented significantly elevated levels of inflammatory and cytokine factors in bronchoalveolar lavage fluid (BALF). Further analysis demonstrated that Hv1 deficiency led to increased malondialdehyde content and decreased superoxide dismutase activity in the BALF of mice exposed to CB particles, indicating increased oxidative stress. Histopathological staining and immunohistochemical experiments confirmed that the absence of the proton channel resulted in thickened alveolar walls, exacerbated inflammatory cell infiltration, and increased fibrous protein deposition in lung tissues. Further immunohistochemical analysis revealed that, compared with WT mice, Hv1 KO mice presented significantly decreased E-cadherin expression and increased vimentin and α-SMA expression in lung tissue after CB particle exposure. Furthermore, exposure to CB particles significantly elevated transforming growth factor-beta 1 levels in the BALF of Hv1 KO mice relative to WT controls. Collectively, these findings demonstrate that Hv1 deficiency potentiates particulate matter-induced lung injury by exacerbating pulmonary inflammation, oxidative stress, and epithelial‒mesenchymal transition. This study establishes Hv1 as a critical protective factor against particulate matter-induced lung damage and highlights its potential as a therapeutic target for preventing and treating particulate matter-associated pulmonary disorders. Graphical abstract

Egg yolk immunoglobulin Y (IgY) has significant application potential in aquaculture as passive immunotherapy against various bacterial infections owing to its capacity for large-scale and cost-effective production. In this study, IgY antibodies of live or inactivated Aeromonas veronii were generated by laying hens immunization. Subsequently, passive immune protection experiments of the two IgY antibodies were conducted on goldfish (Carassius auratus) infected with A. veronii and Aeromonas hydrophila. The results indicated that both live and inactivated bacteria IgY antibodies provided significant passive protection rates (p < 0.05). Furthermore, ELISA tests demonstrated that the two IgY antibodies, as well as the serum of C. auratus, interacted with A. veronii or A. hydrophila (p < 0.05) in vitro. The bacterial loads in the kidneys of C. auratus immunized with the two IgY antibodies were decreased (p < 0.05), and C. auratus phagocytes had enhanced phagocytic activity. The expression levels of antioxidant factors (SOD, CAT, GSH-Px) and inflammatory factors mRNA (TNF-α, IL-1β, IL-6, IL-8) were down-regulated (p < 0.05). Additionally, histopathological analysis indicated that the renal, splenic, and intestinal tissue structures remained intact, and the immunofluorescence confirmed that apoptosis and DNA damage factors of p53 and γH2A.X reduced (p < 0.05), respectively. Thus, the IgY antibodies of live and inactivated A. veronii exhibit passive immune-protective effects against different pathogenic bacteria in C. auratus. Further, inactivated A. veronii immunization causes less damage to laying hens than that of live bacteria, which aligns more closely with welfare standards for laying hens, and the IgY of inactivated A. veronii is anticipated as a cross-protection against A. veronii and A. hydrophila infections in aquaculture.

The white button mushroom (Agaricus bisporus) is a widely cultivated edible and medicinal mushroom, which contains various active substances, and has application value against pathogenic bacteria in aquaculture. Firstly, A. bisporus water extract (AB-WE) was prepared. Through the detection kits, it was found that the polysaccharide, protein, and polyphenol components of AB-WE were 9.11%, 3.3%, and 1.5%, respectively. The 246 compounds were identified in AB-WE, and the major small-molecule components included L-Isoleucine, L-Tyrosine, L-Valine, and Linoleic acid by HPLC-Q Exactive-Orbitrap-MS. Secondly, the AB-WE was evaluated for its immunological activities through dietary administration and pathogen challenge (Aeromonas hydrophila and Vibrio fluvialis) in goldfish (Carassius auratus). The results showed that the levels of immune factors of acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) increased (p < 0.05) in goldfish, and the relative percentage survival of AB-WE against A. hydrophila and V. fluvialis were 80.00% (p < 0.05) and 81.82% (p < 0.05), respectively. The AB-WE reduced the bacterial content in renal tissue, enhanced the phagocytic activity of leukocytes, and exhibited antioxidant and anti-inflammatory effects by reducing the expression of antioxidant-related factors and inflammatory factors. Through histopathological and immunofluorescence techniques, it was found that AB-WE maintained the integrity of visceral tissues and reduced renal tissue apoptosis and DNA damage. Therefore, AB-WE exhibits immunoprotective activity against A. hydrophila and V. fluvialis infections in fish, and holds promise as an immunotherapeutic agent against major pathogenic bacteria in aquaculture.

Egg yolk immunoglobulin Y (IgY) possesses advantages such as low cost, easy availability, simple preparation, high antigen specificity, absence of drug residues, and compliance with animal welfare standards, making it an environmentally friendly and safe alternative to antibiotics. This research utilizes IgY antibody technology to develop a multivalent passive immune vaccine for major pathogenic bacteria in aquaculture. In this study, IgY antibodies against live Shewanella xiamenensis (LSX-IgY) and inactivated S. xiamenensis (ISX-IgY) were prepared by immunizing laying hens, and passive immunization protection experiments were conducted in Carassius auratus infected with S. xiamenensis and Aeromonas hydrophila. The passive immunization protection rates of LSX-IgY and ISX-IgY against S. xiamenensis were 63.64% and 72.73%, respectively, and the passive cross-protection rates against A. hydrophila were 50% and 71.43%, respectively. Further, C. auratus sera could specifically bind to S. xiamenensis or A. hydrophila in vitro, and the phagocytic activity of leukocytes was increased. LSX-IgY and ISX-IgY could reduce the bacterial load in the C. auratus kidneys. Meanwhile, they could significantly reduce the levels of antioxidant factors in serum and inhibit the mRNA expression of inflammation-related factors in the kidneys and spleens. Additionally, histopathology and immunofluorescence analysis showed that both IgY preparations preserved tissue integrity and reduced the expression of apoptosis factor (p53) and DNA damage factor (γH2A.X) of visceral organs, respectively. In summary, LSX-IgY and ISX-IgY can combat various bacterial infections, with no significant difference between the two. Additionally, inactivated bacterial immunization is more aligned with animal welfare standards for laying hens. Therefore, ISX-IgY is expected to serve as a multivalent vaccine against major aquaculture pathogens.