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Using a data set from our laboratory, we assessed the effects of several factors on pig cloning efficiency. The results demonstrated that cells at high confluence (>90%) used as donor cell resulted in higher pregnancy rate, delivery rate and overall cloning efficiency (number of live offspring born per reconstructed embryo transferred to recipients) compared with the cells at 60% to 79% confluence and 80% to 89% confluence. Cells with four, five and six passages compromised the pregnancy and delivery rates compared with first passage cells. The number of blastocysts transferred by somatic cell nuclear transfer (SCNT) did not significantly affect the cloning efficiency, but transfer of blastocyst derived from in vitro culture 5 d after SCNT achieved a significantly higher pregnancy rate compared with one to two cell SCNT embryos from overnight culture. The highest pregnancy rate, delivery rate and the largest litter size were obtained when Bama Miniature pig fibroblasts were used as donor cells and Landrace/Yorkshire hybrid gilts were used as recipients. Recipients treated with chemicals for estrus synchronization had higher pregnancy rates compared with untreated recipients. Our data might be helpful for improving SCNT efficiency in pigs.

[Objective] The structure and function of yak heart fatty acid binding protein 3 (FABP3) gene were analyzed and its expression levels in adult and fetal yak were detected to provide references for xploring biological functions of the gene in yak breeding. [Method] The FABP3 gene was amplified by PCR from heart tissue of Meiren yak, and bioinformatics analysis was carried out after the CDS region was obtained. The expression levels of FABP3 in heart, liver, spleen, lung, kidney, and muscle tissues of adult and fetal yak were detected by real-time quantitative PCR (RT-qPCR). [Result] The length of the coding egion of FABP3 was 402 bp, encoding 133 amino acids. The advanced structure of the protein included mainly β-turn and extended chain. There was no transmembrane region or signal peptide on yak FABP3 protein, indicating it was a stable protein with certain hydrophilicity. Homology analysis of the nucleotide and amino acid sequences of the CDS region showed that the FABP3 gene was conserved in cattle. Accordng to the phylogenetic tree, yak had the closest genetic relationship with wild yak and zebu, and the farhest genetic relationship with chicken. RT-qPCR results showed that the FABP3 gene was expressed in heart, liver, spleen, lung, kidney, and muscle tissues of adult and fetal yak. The expression in liver, spleen, kidney, and muscle of fetal cattle was significantly or extremely significantly higher than that of adult yak, and the expression level in lung of adult yak was significantly higher than that of fetal yak. [Conclusion] The FABP3 gene of yak was cloned, and its tissue expression pattern was explored. This study provided asic data for further study on the role of FABP3 in fat deposition. [ABSTRACT FROM AUTHOR]

Cattle-yak is a hybrid F1 generation of cattle and yak, which has a history of more than 3000 years and has shown better production performance and higher economic benefits than those of yaks. However, up to now, there has been no study on the transcriptome-wide m6A methylation profile of bovine skeletal muscle and its potential biological function during muscle development. Here, we observed significant changes in the expression levels of muscle-related marker genes and methylation-related enzymes during the development of cattle-yak, and the overall m6A content in the Longissimus dorsi muscle of 18-month-old cattle-yak decreased significantly. A total of 36,602 peaks, 11,223 genes and 8388 lncRNAs were identified in the two groups, including 2989 differential peaks (427 up-regulated peaks and 2562 down-regulated peaks), 1457 differentially expressed genes (833 up-regulated genes and 624 down-regulated genes) and 857 differentially expressed lncRNAs (293 up-regulated lncRNAs and 564 down-regulated lncRNAs). GO and KEGG analysis revealed that they were significantly enriched in some muscle-related pathways (Wnt signaling pathway and MAPK signaling pathway) and high-altitude adaptation-related pathway (HIF-1 signaling pathway). Moreover, m6A abundance was positively correlated with gene expression levels, while it was negatively correlated with lncRNA expression levels. This indicates that m6A modification played an important role in the Longissimus dorsi muscle development of cattle-yak; however, the regulation mechanism of m6A-modified mRNA and lncRNA may be different. This study was the first report of transcriptome-wide m6A-modified mRNAs and lncRNAs atlas in the Longissimus dorsi muscle development of cattle-yak, one which will provide new perspectives for genetic improvement in bovines. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the involvement of long non-coding RNA (lncRNA) lung adenocarcinoma transcript 1 (LUADT1) LUADT1 in diabetic retinopathy (DR). We found LUADT1 may interact with miR-383 by RNA interaction prediction. QPCR analysis showed that lncRNA LUADT1 was downregulated, and miR-383 was upregulated in DR. However, correlation analysis revealed no significant correlation between them. In retinal pigment epithelial cells (RPEpiC, h1RPE7 from Sigma-Aldrich), overexpression of LUADT1 and miR-383 failed to affect the expression of each other. However, LUADT1 overexpression led to increased and miR-383 overexpression led to decreased expression level of peroxiredoxin 3 (PRX3). Cell apoptosis analysis showed that LUADT1 and PRX3 overexpression resulted in the decreased cell apoptosis. MiR-383 played an opposite role and reduced the effects of LUADT1 and PRX3 overexpression. Therefore, LUADT1 regulates PRX3 by serving as the endogenous sponge of miR-383 in DR to regulate cell apoptosis. [ABSTRACT FROM AUTHOR]