Hypoxia and low temperature upregulate transferrin to induce hypercoagulability at high altitude.

Studies have shown significantly increased thromboembolic events at high altitude. We recently reported that transferrin could potentiate blood coagulation, but the underlying mechanism for high altitude-related thromboembolism is still poorly understood. Here, we examined the activity and concentration of plasma coagulation factors and transferrin in plasma collected from long-term human residents and short-stay mice exposed to varying altitudes. We found that the activities of thrombin and factor XIIa (FXIIa) along with the concentrations of transferrin were significantly increased in the plasma of humans and mice at high altitudes. Furthermore, both hypoxia (6% O2) and low temperature (0°C), 2 critical high-altitude factors, enhanced hypoxia-inducible factor 1α (HIF-1α) levels to promote the expression of the transferrin gene, whose enhancer region contains HIF-1α binding site, and consequently, to induce hypercoagulability by potentiating thrombin and FXIIa. Importantly, thromboembolic disorders and pathological insults in mouse models induced by both hypoxia and low temperature were ameliorated by transferrin interferences, including transferrin antibody treatment, transferrin downregulation, and the administration of our designed peptides that inhibit the potentiation of transferrin on thrombin and FXIIa. Thus, low temperature and hypoxia upregulated transferrin expression-promoted hypercoagulability. Our data suggest that targeting the transferrin-coagulation pathway is a novel and potentially powerful strategy against thromboembolic events caused by harmful environmental factors under high-altitude conditions.

Ovary Transcriptome Profiling in Broody and Egg-laying Chahua Chickens.

Broodiness in egg-laying hens (EHs) leads to ovarian atrophy, resulting in reduced egg-laying performance. However, the ovarian regulatory mechanisms in broody hens (BCs) remain elusive. Therefore, ovaries were removed from 300-day-old BCs and EHs for RNA sequencing. Ovarian morphology and histological characteristics of the BC and EH groups were compared and analyzed. The EH group had significantly more hierarchical follicles (HFs) and small yellow follicles (SYFs) than that of the BC group. Although several secondary follicles (SFs) and primary follicles were observed in the ovaries of the EH group, only a few SFs were observed in the ovaries of the BC group. Subsequently, RNA-sequencing analysis was conducted to determine the ovarian expression profiles of the two groups. Transcriptome sequencing identified 259 differentially expressed genes (DEGs) between the BC and EH groups. Of the 259 DEGs, 136 were upregulated and 123 were downregulated. The DEGs were mapped to 22 gene ontology terms and 4 Kyoto Encyclopedia of Genes and Genomes pathways for ovarian tissue. The analysis showed that matrix metalloproteinases 11/13 (MMP11/MMP13) were enriched in the extracellular matrix. The extracellular matrix mediated by MMP13 is affected by follicle-stimulating hormone, prolactin, and estrogen, which are critical signaling pathways that may affect ovarian follicle development to regulate the large yellow follicle reserve process and the ovulation cycle of broody Chahua chickens. These findings indicate that understanding differences in gene expression between the ovarian tissues of BCs and EHs could serve as a valuable reference point for enhancing egg-laying performance in Chahua chickens.

Effect of maternal lactoferrin supplementation on iron contents and anti-oxidant capacity in Dahe black Pig neonates.

Iron levels are closely related to animals' growth performance and anti-oxidant function. Lactoferrin (LF) is an iron-binding glycoprotein, which can promote the absorption of iron and regulate immune function. This study aimed to clarify the effect of maternal LF supplementation on the iron metabolism of Dahe piglets. Sixty sows (Dahe black, parity 3-4, no significant differences in body weight) were randomly assigned to five groups: control (basal diet with no iron supplementation), supplemented 100 (LF1 group), 200 (LF2 group), or 300 (LF3 group) mg LF/kg in the basal diet, and the basal diet supplemented with 100 (Fe-Gly group) mg Fe/kg as ferrous glycine (Fe-Gly). The serum anti-oxidant parameters of the sows and neonatal piglets were determined. The iron contents, anti-oxidant gene expression levels, and Fe-acquisition genes were detected in the liver, heart, spleen, and other neonatal organs. The results indicated that (1) the LF3 group of sows had the highest serum and colostrum iron contents (P < 0.05). The maternal LF significantly promoted the iron stores in the heart, liver, spleen, and lung of piglets compared with Fe-Gly. (2) The maternal LF increased serum glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) activities of sows. Compared with other groups, the total anti-oxidant capacity (T-AOC) activity of LF2 groups increased significantly (P < 0.05). (3) LF significantly increased piglet serum GSH-Px, T-SOD, and T-AOC activities (P < 0.05). (4) Gene expression levels of GSH-Px, and SOD in the duodenum and jejunum of the LF2 group were significantly higher than in the Fe-Gly group (P < 0.05), while the expression levels in the liver and heart were lower (P < 0.05). (5) The expression levels of hepcidin and LF in the liver and duodenum of the LF2 group were significantly higher than in the Fe-Gly group (P < 0.05). In conclusion, maternal LF supplementation showed remarkable effects on iron storage in neonatal piglets, and exhibited strong antioxidant activities, it is helpful to prevent the occurrence of iron deficiency, and improves the immune function of animals.

Correlation analysis of muscle amino acid deposition and gut microbiota profile of broilers reared at different ambient temperatures.

OBJECTIVE: Temperature could influence protein and amino acid deposition as well as gut microbiota profile and composition. However, the specific effects of ambient temperature on amino acids deposition and gut microbiota composition remain insufficiently understood. METHODS: A total of 300 one-day-old Avian broilers were randomly divided into three groups and reared at high, medium, and low temperature (HT, MT, and LT), respectively. Breast muscle and fecal samples were collected for amino acid composition analysis and 16S rRNA gene sequence analysis. RESULTS: Our data showed that compared to the MT group, there was a decrease of muscle leucine and tyrosine (p<0.05), as well as an increase of methionine in the HT group (p<0.05) and a decrease of serine in the LT group. Examination of microbiota shift revealed that at genus level, the relative abundance of Turicibacter and Parabacteroides was increased in the HT group (p<0.05) and that the relative abundances of Pandoraea, Achromobacter, Prevotella, Brevundimonas, and Stenotrophomonas in the LT group were higher than those in the MT group (p<0.05). In addition, there were substantial correlations between microbes and amino acids. In the HT group. Turicibacter was negatively correlated with aspartic acid and tyrosine, whereas Parabacteroides was positively correlated with methionine (p<0.05). In the LT group, there were multiple positive correlations between Achromobacter and arginine, isoleucine or tyrosine; between Prevotella and cysteine or phenylalanine; between Brevundimonas and cysteine; and between Stenotrophomonas and cysteine as well as a negative correlation between Stenotrophomonas and serine. CONCLUSION: Our findings demonstrated that amino acid content of breast muscle and intestinal microbiota profile was affected by different ambient temperatures. Under heat exposure, augmented abundance of Parabacteroides was correlated with elevated methionine. Low temperature treatment may affect muscle tyrosine content through the regulation of Achromobacter.

[Effect of interactions of chromatin regulatory elements with different promoters on the regulation of gene expression].

The effect of altering the promoter region of ubiquitous chromatin-opening element (UCOE) and matrix attachment region (MAR) on stable and efficient expression of genes was investigated. Four different promoters were tested, namely, oct4 containing an enhancer region, sox2 having a CpG island, nanog having no regulatory elements, and CMV containing a CpG island and an enhancer region. Eight reporter plasmids were constructed: pOCT4-UCOE, pOCT4-MAR, pSOX2-UCOE, pSOX2-MAR, pNANOG-UCOE, pNANOG-MAR, pCMV-UCOE, and pCMV-MAR. Stable and efficient expression was observed when UCOE combined with the oct4 promoter, whereas the sox2 was the best promoter suited for MAR. Comparison of the stable clones of oct4-UCOE and sox2-MAR showed that UCOE-regulated expression is more stable and efficient than MAR-regulated expression. When CpG island-containing promoter is linked with UCOE, stable and efficient expression could be observed. These data suggest that an enhancer region in the promoter leads to high, yet unstable expression when combined with UCOE, whereas CpG islands stabilize expression. In conclusion, UCOE and MAR interact with regulatory elements on the promoter by altering the chromatin open state and chromatin loop to regulate gene expression.