Liangpeng Ge

Background: Pigs are economically critical livestock andan optimal model for investigating the development and diversification of antibody repertoires. The Rongchang (RC) pig, a nationally protected indigenous breed in China, possesses unique genetic characteristics, yet genomic-level research on its antibody repertoire remains limited, as most porcine antibody studies have focused on Landrace pigs. Methods: To decipher the genetic features of the antibody library in RC pigs, we used immunogenomic high-throughput sequencing to systematically analyze the antibody repertoires of five healthy purebred pig breeds, including two indigenous breeds (RC and BM) and three commercial breeds (Yorkshire, Duroc, and Landrace), with a focus on comparing conserved patterns and breed-specific differences in V(D)J gene utilization between Rongchang pigs and the other four breeds. Results: All five breeds exhibited a strong preference for a conserved subset of core IGHV genes. Notably, this study detected functional IGHJ3 expression (0.40.8%) in all examined breeds, contradicting the conventional view that only IGHJ5 is functional in porcine antibody repertoires. Among them, RC pigs showed the lowest IGHJ3 frequency. Furthermore, RC pigs ranked second in antibody repertoire diversity among the five breeds, implying abundant antigen-binding specificity, and exhibited the lowest CDR3 proportion, indicating breed-specific V(D)J recombination preferences. Conclusions: These results clarify the conserved and breed-specific characteristics of RC pig antibody repertoires, establishing a basis for exploring the genetic regulation of V(D)J-mediated antibody repertoire changes under varied immune conditions. This work also provides genomic support for the rational utilization of RC pig genetic resources.

The growing need for rapid and cost-effective antibody production highlights the potential of Komagataella phaffii (syn. Pichia pastoris) as an alternative

The commensal microbiota provides immunomodulatory signals during the development, differentiation and activation of immune cells, and is crucial for maintaining host immune homeostasis. However, the systematic effects of commensal microbiota on host immunity based on large animal model at the single-cell level remain to be resolved. Here, we utilized single-cell RNA sequencing (scRNA-seq) to analyze the transcriptome profiling containing 57,720 cells from three important immune tissues [Peyer’s patches (PP), mesenteric lymph node (MLN), and spleen] of germ-free (GF) and specific pathogen-free (SPF) piglet. We presented detailed description of the dataset and preliminarily identified the major cell types including immune and non-immune cells, and further annotated the immune cell subsets. This dataset provides a data mining resource for researchers involved in microbe-host interactions, and enables in-depth analysis of cell map alterations caused by the microbiota colonization during early immune development. As the first single-cell transcriptomics dataset for immune tissue of GF and SPF piglet, this provides a valuable data resource for the study of commensal microbe-host immunity regulation.

Tendons connect animal skeletons to skeletal muscles, playing a crucial role in weight-bearing and maintaining motor functions. After decellularization, tendon extracellular matrix (tECM) retains the physicochemical characteristics similar to those of native tendons. This has made tECM a promising biomaterial in the fields of tissue engineering and regenerative medicine in recent years. This paper summarizes the origin, structure, and ECM components of animal tendons, reviews decellularization methods, and discusses recent advancements in the research and applications of decellularized tendons. Furthermore, it explores future development trends of xenogeneic decellularized tendon materials, aiming to provide a reference for fundamental research and the development of biomaterials related to decellularized tendons.

The gut microbiota plays a crucial role in the growth performance, health status, and welfare of pigs. Breast milk is a key factor in the colonization of gut microbiota and the overall health of newborn piglets. With advancements in breeding technology, formula milk has been widely adopted as a substitute for breast milk. This study aims to investigate the effects of sow feeding (natural breastfeeding) and formula milk feeding on the gut microbiota of specific pathogen-free (SPF) Bama pigs. Using metagenomic sequencing technology, we analyzed 114 fecal samples to uncover the impacts of different feeding methods on gut microbial diversity, dominant microbial populations, metabolic functions, carbohydrate-active enzymes (CAZymes), and antibiotic resistance genes (ARGs). The results revealed significant differences in the structure and function of gut microbiota between the breast milk (BM) group and the formula milk (FM) group at day 21. The BM group exhibited higher gut microbial diversity compared to the FM group, along with more extensive metabolic functions at both the gene and species levels. Notably, the FM group demonstrated higher activity in galactose metabolism and glycan metabolism, particularly at day 21. Additionally, the FM group showed significantly higher levels of ARGs against glycopeptide antibiotics at days 21 and 28 compared to the BM group. This study also found that breastfeeding and formula feeding differentially regulate the metabolic activity of gut microbiota and the expression of related enzymes, which may have long-term effects on nutrient absorption and disease resistance in pigs. These findings provide new insights into how different feeding methods shape the gut microbiota of pigs and offer a scientific basis for optimizing feeding strategies and improving breeding efficiency.

Neonatal mammals must rapidly adapt to significant physiological changes during the transition from the intrauterine to extrauterine environments. This adaptation, particularly in the metabolic and respiratory systems, is essential for survival. MicroRNAs (miRNAs) are small noncoding RNAs that regulate various physiological and pathological processes by binding to the 3′ untranslated regions of mRNAs. This study aimed to identify miRNAs involved in the early extrauterine adaptation of neonatal piglets and explore their functions. We performed small RNA sequencing on six tissues (heart, liver, spleen, lung, multifidus muscle, and duodenum) from piglets 24 h before birth (day 113 of gestation) and 6 h after birth. A total of 971 miRNA precursors and 1511 mature miRNAs were identified. Tissue-specific expression analysis revealed 881 tissue-specific miRNAs and 164 differentially expressed miRNAs (DE miRNAs) across the tissues. Functional enrichment analysis showed that these DE miRNAs are significantly enriched in pathways related to early extrauterine adaptation, such as the NFκB, PI3K/AKT, and Hippo pathways. Specifically, miR-22-3p was significantly upregulated in the liver post-birth and may regulate the PI3K/AKT pathway by targeting AKT3, promoting gluconeogenesis, and maintaining glucose homeostasis. Dual-luciferase reporter assays and HepG2 cell experiments confirmed AKT3 as a target of miR-22-3p, which activates the AKT/FoxO1 pathway, enhancing gluconeogenesis and glucose production. Furthermore, changes in blood glucose and liver glycogen levels in newborn piglets further support the role of miR-22-3p in glucose homeostasis. This study highlights the importance of miRNAs, particularly miR-22-3p, in the early extrauterine adaptation of neonatal piglets, offering new insights into the physiological adaptation of neonatal mammals.

Ebola disease is a lethal viral hemorrhagic fever caused by ebolaviruses within the Filoviridae family with mortality rates of up to 90%. Monoclonal antibody (mAb) based therapies have shown great potential for the treatment of EVD. However, the potential emerging ebolavirus isolates and the negative effect of decoy protein on the therapeutic efficacy of antibodies highlight the necessity of developing novel antibodies to counter the threat of Ebola. Here, 11 fully human mAbs were isolated from transgenic mice immunized with GP protein and recombinant vesicular stomatitis virus-bearing GP (rVSV-EBOV GP). These mAbs were divided into five groups according to their germline genes and exhibited differential binding activities and neutralization capabilities. In particular, mAbs 8G6, 2A4, and 5H4 were cross-reactive and bound at least three ebolavirus glycoproteins. mAb 4C1 not only exhibited neutralizing activity but no cross-reaction with sGP. mAb 7D8 exhibited the strongest neutralizing capacity. Further analysis on the critical residues for the bindings of 4C1 and 8G6 to GPs was conducted using antibodies complementarity-determining regions (CDRs) alanine scanning. It has been shown that light chain CDR3 played a crucial role in binding and neutralization and that any mutation in CDRs could not improve the binding of 4C1 to sGP. Importantly, mAbs 7D8, 8G6, and 4C1 provided complete protections against EBOV infection in a hamster lethal challenge model when administered 12 h post-infection. These results support mAbs 7D8, 8G6, and 4C1 as potent antibody candidates for further investigations and pave the way for further developments of therapies and vaccines.

Background: Angiogenesis is essential for various physiological and pathological processes, such as embryonic development and cancer cell proliferation, migration, and invasion. Long noncoding RNAs (lncRNAs) play pivotal roles in normal homeostasis and disease processes by regulating gene expression through various mechanisms, including competing endogenous RNAs (ceRNAs) of target microRNAs (miRNAs). The lncRNA MYU is known to promote prostate cancer proliferation via the miR-184/c-Myc regulatory axis and to be upregulated in vascular endothelial cells under hypoxic conditions, which often occurs in solid tumors. In the present study, we investigated whether MYU might affect cancer growth by regulating angiogenesis in vascular endothelial cells under hypoxia. Methods: The expression of MYU-regulated miR-23a-3p and interleukin-8 (IL-8) in HUVEC cell lines was examined using qRT-PCR. The CCK-8 assay, EdU assay, wound-healing assay, and tube-formation assay were used to assess the effects of MYU on cell proliferation, migration, and tube formation of HUVEC cells in vitro. The dual-luciferase reporter assay was performed to examine the effects of miR-23a-3p on MYU and IL-8 expression. Results: We found that the overexpression of MYU and knockdown of miR-23a-3p in human umbilical vein endothelial cells (HUVECs) under hypoxia promoted cell proliferation, migration, and tube formation. Mechanistically, MYU was shown to bind competitively to miR-23a-3p, thereby preventing miR-23a-3p binding to the 3′ untranslated region of IL-8 mRNA. In turn, increased production of pro-angiogenic IL-8 promoted HUVEC proliferation, migration, and tube formation under hypoxia. Conclusion: This study identified a new role for lncRNA MYU as a ceRNA for miR-23a-3p and uncovered a novel MYU–miR-23a-3p–IL-8 regulatory axis for angiogenesis. MYU and/or miR-23a-3p may thus represent new targets for the treatment of hypoxia-related diseases by promoting angiogenesis.