The purpose of this study was to establish an efficient pig cell gene editing technology system and lay a foundation for the application of PE system in pig genetics and breeding. Multiple candidate sites were selected from the coding region of VIM gene， and sgRNA was designed strictly according to the target site design principle of CRISPR/Cas9 system. The candidate sgRNAs were scored by bioinformatics tools， and the two sgRNA sequences with the highest scores were screened out. Based on the above two sgRNAs， the corresponding pegRNAs were further designed. The cells were treated with drugs hydroxyurea and nocodazole to stay in the G1/S or G2/M phase， and the nCas9-RT fusion protein editor vector and pegRNA expression vector were electro-transfected into the cells. After 48 h， the genomic DNA of the mixed cells was extracted， the target sites were amplified by PCR， and the next-generation sequencing was performed. CRISPResso2 was used to analyze the editing results. The results showed that the editing efficiency of PE2 system in PK15 cells was between 0.16% and 14.03%. The editing efficiency in IPEC-J2 cells was between 0.08% and 0.58%. The editing efficiency of PEmax system in PK15 cells was between 0.63% and 23.25%. However， the editing efficiency of PE4 and PE4max systems in PK15 cells was poor， with the highest editing efficiency of 0.7% and 0.36%， respectively. Regulating the cell cycle could improve the editing efficiency of the PE2 system for the editing types of single base substitution and single base deletion of porcine VIM gene. The editing efficiency of the former had increased by 1.4 times， and the editing efficiency of the latter had increased by 2.6 times. According to the results， PE technology can perform a variety of different types of precise editing of pig VIM gene and improve its editing efficiency through cell cycle regulation. It provides a technical reference for the next step to combine somatic cell cloning technology to prepare pig populations with improved production performance.

This study aimed to investigate the effects of Escherichia coli （E. coli）， Clostridium butyricum （C. butyricum）， and their metabolites on mitochondrial function and related metabolic pathways in porcine intestinal epithelial cells （IPEC-J2）， providing a theoretical basis for regulating intestinal health. Initially， bacterial suspensions were screened to ensure logarithmic-phase growth （3 h） with optimal viability for subsequent experiments. IPEC-J2 cells were then treated with varying concentrations of Escherichia coli， Clostridium butyricum， lipopolysaccharide （LPS）， and short-chain fatty acids （SCFAs）， with 3 replicates per group. Cell viability and morphological integrity were assessed to determine the optimal treatment duration and concentration. Key metrics included ATP levels， reactive oxygen species （ROS）， mitochondrial membrane potential， and the expression of genes related to antioxidant defense and mitochondrial function. The results showed that： 1） The optimal concentrations for treating IPEC-J2 cells were determined as 10⁸ CFU·mL^-1 for Escherichia coli and 10⁶ CFU·mL^-1 for C. butyricum， with a treatment duration of 1 h. The treatment duration of SCFAs and LPS were also treated for 1 h. 2） Propionic acid and E. coli significantly increased ROS levels （P<0.05）， whereas C. butyricum and LPS reduced them （P<0.05）. 3） C. butyricum upregulated the expression of ACC1， NRF1， TFAM， HO-1， APAF-1， PIG3， and p53 （P<0.05）； E. coli downregulated the expression of TFAM and CtyC （P<0.05） but upregulated the expression of p53 （P<0.05）； Propionic acid and acetic acid had no significant effects on genes expression. The results indicated that SCFAs enhanced cellular metabolism， E. coli and LPS triggered oxidative stress and apoptotic pathways. C. butyricum activated mitochondrial lipid metabolism， antioxidant defenses， and apoptosis in IPEC-J2 cells. These findings offer novel insights into mitochondria-targeted strategies for gut microbiota regulation.

This study aimed to estimate the genetic parameters and genomic breeding values of “Min Gai” cashmere goat hair traits， compare the effects of GBLUP and Bayes methods on the accuracy of genomic breeding value prediction of hair traits of “Min Gai” cashmere goat， and provide a reference basis for optimising the genetic selection and breeding strategy of “Min Gai” cashmere goats. This study took the fleece traits of 847 “Min Gai” cashmere goats as the research object and used GBLUP， Bayes A， Bayes B， Bayes C， and Bayes LASSO to estimate the genetic parameters and genomic breeding values of fleece traits （fleece length， cashmere length， cashmere diameter， and cashmere yield）. Furthermore， the accuracy of the genomic breeding value estimates was evaluated using five-fold cross-validation. The results showed that the heritability of fleece length estimated by the 5 methods was 0.11-0.32， the heritability of cashmere length was 0.14-0.33， the heritability of cashmere diameter was 0.17-0.49， and the heritability of cashmere yield was 0.16-0.32. There was a strong positive genetic correlation between cashmere length and fleece length， with a correlation coefficient of 0.48-0.84. The genetic correlation between other traits was relatively small， ranging from -0.06 to 0.46. The genomic breeding value accuracy estimated by the GBLUP， Bayes A， Bayes B， Bayes C， and Bayes LASSO methods for fleece length， cashmere length， cashmere diameter， and cashmere yield were 0.623 4-0.950 9， 0.626 2-0.953 0， 0.830 8-0.953 1， and 0.741 8-0.961 4， respectively. The genomic prediction accuracy of each trait estimated by the Bayes method was significantly higher than that estimated by the GBLUP method， and the Bayes B method had the highest genomic prediction accuracy， reaching 0.950 9-0.961 4. In summary， the Bayes B method is recommended for the genomic selection of “Min Gai” cashmere goats， which may accelerate the genetic progress of the population.

This study aimed to develop a haplotype-segment matching-based maternal grandsire discovery approach applicable to different SNP array densities and segment window sizes in the absence of dam genotypes， thereby providing a reliable technical approach for pedigree reconstruction. The study utilized genomic data from 6 117 dairy cows to construct datasets of varying SNP densities， including low-density （5K， 10K） and medium-density （20K， 50K） SNP panels. For each dataset， the genome was partitioned into fixed-length haplotype segments of 5， 10， 35， 75， and 120 SNPs， followed by haplotype inference. Based on the known genotypes of the offspring's sire， paternal haplotypes were excluded to infer maternal haplotypes， which were then compared with those of candidate bulls on a segment-by-segment basis to calculate the haplotype matching rate （HMR）. The bull with the highest HMR was designated as the putative maternal grandsire （MGS）. A total of 345 individuals with complete and validated pedigree information were used to verify method accuracy. Furthermore， genomic relatedness coefficients and homozygosity levels were calculated to investigate the potential effects of kinship structure and inbreeding on identification outcomes. The haplotype-segment matching method achieved more than 90% assignment accuracy across most combinations of SNP-array density and segment window length. The 20K array with a 35-SNPs window yielded the highest accuracy at 98%， demonstrating robust performance. Nonetheless， the optimal decision thresholds for the haplotype match rate and the resulting accuracies varied substantially with parameter settings， indicating that SNP density and window length were key determinants of performance. Additional analyses showed that kinship structure and inbreeding levels significantly affected matching performance and assignment accuracy， suggesting that the population genetic background should be considered in practical applicatons. The haplotype-segment matching （HSM）-based method demonstrated high accuracy in maternal grandsire identification under conditions of missing maternal genotype data， making it a valuable tool for pedigree correction in large-scale commercial herds. Optimizing SNP density and haplotype window length based on the genetic background of the population is essential to enhance identification efficiency and robustness in practical applications.

This study aimed to systematically identify expression short tandem repeats （eSTRs） in dairy cattle that are significantly associated with gene expression， in order to explore their regulatory roles in gene expression and complex economic traits， thereby providing functional variant resources for molecular breeding. A total of 108 Chinese Holstein cows were sampled for tail vein blood collection. Genomic DNA and total RNA were extracted and subjected to 30×whole-genome resequencing and transcriptome sequencing， respectively. After quality control， 105 samples were retained for further analysis. STRs were annotated and genotyped at the population scale using TRF and HipSTR. Gene expression levels were quantified after quality filtering with Trimmomatic， alignment using STAR， and normalization with edgeR. Based on a ±1 Mb window around transcription start sites （TSS）， a linear model was used to assess the association between STR dosage and gene expression to identify cis-eSTRs. Weighted correlation network analysis （WGCNA） and Gene Ontology （GO） were further applied to explore the functional relevance of cis-eSTRs in complex traits. A total of 25 154 significant cis-eSTR-gene expression associations were identified. These cis-eSTRs were predominantly located in intronic regions and near TSSs. WGCNA revealed that cis-eSTR regulated genes clustered into several functional modules， some of which were significantly associated with traits related to milk production and immune. GO enrichment analysis revealed that these modules are broadly involved in biological processes such as immune response， lipid metabolism， cell cycle， DNA replication and stability. Several key genes （e.g.， IRF7， BoLA-DRB， CCT2） might play important functional roles in immune regulation and mammary gland function. cis-eSTRs play important roles in regulating gene expression and complex traits in dairy cattle. The cis-eSTR resource generated in this study provides a valuable foundation for the functional annotation of complex traits and offers potential molecular markers for breeding applications.

The aim of this study was to explore the impact of molecular networks constructed by DE-circRNAs related to milk fat synthesis on milk fat synthesis in dairy cows through in-depth analysis. In this study， we used the circRNA library and transcriptome sequencing data of Holstein dairy cows with high and low milk fat percentage constructed previously to screen and identify DE-circ RNAs（circPRKAA2） related to milk fat synthesis， and further predicted their ceRNA regulatory network using Targetscan and miRanda software. Real-time fluorescence quantitative PCR， RNA pull down assay and dual luciferase reporter assay were used to verify the targeting relationship. Oil red O and BODIPY staining were used to investigate the effect of target miRNA on lipid droplet secretion in BMECs. The results indicated that the regulatory relationship of the ceRNA （circPRKAA2/miR-22-3p/SLC27A1） network was successfully predicted and verified， and 9 miRNAs and 10 candidate mRNAs were predicted， and it was clarified that circPRKAA2 and miR-22-3p were mainly distributed in the cytoplasm of BMECs. circPRKAA2 negatively regulated miR-22-3p during the process of milk fat synthesis in BMECs. miR-22-3p inhibited the secretion of lipid droplets， the production of triglycerides and cholesterol in BMECs， and negatively regulated SLC27A1. In conclusion， this study analyzed and verified the targeting relationship of circPRKAA2/miR-22-3p/SLC27A1 and its regulatory effect on milk fat synthesis， with the hope of providing new insights for improving milk quality.

The aim of this study was to investigate the differential expression of genes in the blood of Yili horses before and after the 5 000 m race， and the correlation between cardiac parameters and the differentially expressed genes. Six elite Yili horses， which were among the top 8 horses in the 5 000 m speed race at the national level， were selected for the study. Echocardiograms were collected from the resting state of the horses before the 5 000 m speed race and the blood of the horses before and after the race by using the Myriad M6 Veterinary Portable Color Doppler Ultrasound System， which was used for transcriptome sequencing. The results showed that： 1） A total of 889 differentially expressed genes were screened， including 751 up-regulated genes and 138 down-regulated genes.GO and KEGG enrichment analyses revealed that the differentially expressed genes were mainly involved in the biological processes of cellular response to stimuli and immune regulation， as well as the MAPK and Jak-STAT signaling pathways involved in cellular immune response and inflammation. 2） CCL3， CSF1， CSF3， IFNG， IL4， IL10 were screened as key genes. In the correlation between the post-competition expression of key genes and cardiac structural indexes， CCL3 was extremely significantly positively correlated with PADs （P<0.01）， significantly positively correlated with LVIDd （P<0.05）， and significantly negatively correlated with RWTd （P<0.05）， CSF1 was extremely significantly positively correlated with PADd （P<0.01）， significantly negatively correlated with RWTd （P<0.05），CSF3 was significantly negatively correlated with IVSd and EDV （P<0.05）， IFNG was significantly positively correlated with PADd （P<0.05）， significantly negatively correlated with IVSd， MWTd， and RWTd （P<0.05）， IL10 was extremely significantly positively correlated with LVLD （P<0.01）， significantly positively correlated with MVD （P<0.05）， and significantly negatively correlated with EDV （P<0.05）. In the correlation between the post-race expression of key genes and cardiac function indexes， CSF3 was significantly negatively correlated with SV， SI， CI， and VTI （P<0.05）， IFNG was significantly negatively correlated with SI （P<0.05）， and IL10 was significantly negatively correlated with CO， CI， and VTI （P<0.05）. Moreover， CSF1 and IFNG post-race expression were significantly negatively correlated with the 5 000 m exercise performance of Yili horses （P<0.05）. Cardiac indexes and CCL3， CSF3， IL4， IL10 regulatory mechanisms play important roles in the 5 000 m race of Yili horses， and the expression levels of CSF1 and IFNG can be used as candidate genes to assess the athletic performance of Yili horses.

This study aimed to investigate the anti-aging effects and underlying molecular mechanisms of reserve mesenchyme cells （RMCs） derived from the velvet antler of the sika deer on hydrogen peroxide-induced senescent human dermal fibroblasts （HDFs）. An HDF senescence model was established by treatment with 200 µmol·L^-1 H₂O₂ and confirmed via SA‑β‑galactosidase staining. Senescent HDFs were then co‑cultured with RMCs， and subsequent assays evaluated cell proliferation， migration， superoxide dismutase （SOD） activity， and malondialdehyde （MDA） content. The 4D-DIA proteomics technology was used to identify differential proteins between co-culture medium and senescent HDF medium， and bioinformatics enrichment analysis was performed. Functional analyses revealed that， compared with the model group， RMCs co‑culture significantly enhanced HDFs proliferation and migration （P<0.01）， increased SOD activity （P<0.05）， and decreased MDA levels （P<0.05）. Using 4D‑DIA proteomics， 289 differentially expressed proteins were identified in the conditioned media of the co‑culture versus senescent HDF alone， with key molecules such as ATP1A1， COL7A1， IGF2R， and MMP3 showing significant expression changes. KEGG enrichment analysis highlighted the proteasome， PPAR signaling pathway， extracellular matrix-receptor interaction， and focal adhesion pathways as principal regulatory axes. These findings suggest that velvet-antler-derived RMCs from the sika deer alleviate oxidative stress-induced dysfunction in senescent HDFs. It was speculated that RMCs may exert their effects by enhancing proteasome‑mediated clearance of damaged proteins， activating PPARγ to suppress inflammation and oxidative stress， promoting COL7A1‑dependent dermal-epidermal junction reconstruction， and inhibiting MMP3‑mediated extracellular matrix degradation， thus maintaining HDF homeostasis and function. This work provides experimental evidence and a theoretical foundation for developing deer antler-derived secretome-based strategies for skin anti‑aging.

This study aimed to investigate the impact of heat stress on the antioxidant capacity of porcine sertoli cells and to further elucidate the protective role and underlying mechanism of resveratrol （RES） in alleviating oxidative stress. Porcine sertoli cells were isolated and cultured primarily. The cells were divided into 3 groups： control group （NC， 34 ℃）， heat stress group （HS， 43 ℃ for 30 min）， and heat stress+resveratrol group （HS+200 nmol·L^-1 RES）. The levels of reactive oxygen species （ROS）， the content of malondialdehyde （MDA） and advanced glycation end products （AGEs）， as well as the activities of superoxide dismutase （SOD） and catalase （CAT） were measured. The expression levels of RAGE， NOX4， and NF-κB genes were determined using real-time fluorescence quantitative PCR. Compared with the control group， heat stress significantly increased ROS level （P<0.05）， MDA content （P<0.05）， and AGEs content （P<0.01）， whereas the activities of SOD and CAT were significantly decreased （P<0.01）， indicating that heat stress successfully induced oxidative stress in porcine sertoli cells. Pretreatment with 200 nmol·L^-1 RES significantly attenuated the heat stress-induced elevations in ROS， MDA， and AGEs levels （P<0.05）， and restored the activities of SOD and CAT （P<0.05）. Moreover， RES significantly downregulated the expression levels of RAGE， NOX4， and NF-κB genes induced by heat stress （P<0.05）. Heat stress disrupts redox homeostasis in porcine sertoli cells， leading to oxidative stress. Resveratrol alleviates this heat stress-induced oxidative stress by suppressing the AGEs-RAGE/NOX4/NF-κB signaling pathway. The findings provide a theoretical basis for utilizing resveratrol to improve boar semen quality during summer.

The objective of this study was to investigate the imprinting status and to search for potential DNA differentially methylated regions （DMRs） of the DLK1-DIO3 domain in bovine placentas. In this study， whole genome sequencing （WGS） was performed on 3 bovine placentas and their parents to mining the informative single nucleotide polymorphisms （SNPs）. The imprinting status of genes in the DLK1-DIO3 domain were determined by the combined analysis of WGS and transcriptome sequencing （RNA-seq） results. The DMRs were identified by combination of WGS and whole genome bisulfite sequencing （WGBS） data. WGS and RNA-seq results showed that MEG3， MEG8 and MEG9 genes were maternally expressed imprinted genes and DLK1， RTL1 and DIO3 genes were paternally expressed imprinted genes. The WGBS data analysis showed that 4 DMRs， Dlk1-DMR， IG-DMR， Gtl2-DMR and Meg8-DMR were conserved in human and mouse， and 5 new DMRs were identified， DLK1-DMR0 and DLK1-DMR1 located upstream of the DLK1 gene， MEG9-DMR in MEG9 promoter region， and DIO3-DMR1 and DIO3-DMR2 located upstream of DIO3 genes， respectively. The results indicate that the imprinted status and differentially methylated regions of bovine DLK1-DIO3 imprinted region are highly conserved to human and mouse. In addition， 5 potential DMRs were identified in bovine DLK1-DIO3 region， and their regulating mechanism on the DLK1-DIO3 imprinted region needs to be further investigated.