Functional analysis of differentially expressed circular RNAs in sheep subcutaneous fat.

BACKGROUND: Circular RNAs (circRNAs), as important non-coding RNAs (ncRNAs), are involved in many biological activities. However, the exact chemical mechanism behind fat accumulation is unknown. In this paper, we obtained the expression profiles of circRNAs using high-throughput sequencing and investigated their differential expression in subcutaneous fat tissue of Duolang and Small Tail Han sheep. RESULTS: From the transcriptomic analysis, 141 differentially expressed circRNAs were identified, comprising 61 up-regulated circRNAs and 80 down-regulated circRNAs. These host genes were primarily enriched in the MAPK and AMPK signaling pathways which is closely associated with fat deposition regulation. We identified circRNA812, circRNA91, and circRNA388 as vital genes in fat deposition by miRNA-circRNA target gene prediction. The functional annotation results of target genes of key circRNAs showed that the signaling pathways mainly included PI3K-Akt and AMPK. We constructed the competing endogenous RNA (ceRNA) regulatory network to study the role of circRNAs in sheep lipid deposition, and circRNA812, circRNA91, and circRNA388 can adsorb more miRNAs. NC_040253.1_5757, as the source of miRNA response element (MRE) among the three, may play an important role during the process of sheep fat deposition. CONCLUSIONS: Our study gives a systematic examination of the circRNA profiles expressed in sheep subcutaneous fat. These results from this study provide some new basis for understanding circRNA function and sheep fat metabolism.

New Dawn for Atherosclerosis: Vascular Endothelial Cell Senescence and Death.

Endothelial cells (ECs) form the inner linings of blood vessels, and are directly exposed to endogenous hazard signals and metabolites in the circulatory system. The senescence and death of ECs are not only adverse outcomes, but also causal contributors to endothelial dysfunction, an early risk marker of atherosclerosis. The pathophysiological process of EC senescence involves both structural and functional changes and has been linked to various factors, including oxidative stress, dysregulated cell cycle, hyperuricemia, vascular inflammation, and aberrant metabolite sensing and signaling. Multiple forms of EC death have been documented in atherosclerosis, including autophagic cell death, apoptosis, pyroptosis, NETosis, necroptosis, and ferroptosis. Despite this, the molecular mechanisms underlying EC senescence or death in atherogenesis are not fully understood. To provide a comprehensive update on the subject, this review examines the historic and latest findings on the molecular mechanisms and functional alterations associated with EC senescence and death in different stages of atherosclerosis.

Genome-Wide Analysis of microRNAs Identifies the Lipid Metabolism Pathway to Be a Defining Factor in Adipose Tissue From Different Sheep.

microRNAs are a class of important non-coding RNAs, which can participate in the regulation of biological processes. In recent years, miRNA has been widely studied not only in humans and mice, but also in animal husbandry. However, compared with other livestock and poultry breeds, the study of miRNA in subcutaneous adipose tissue of sheep is not comprehensive. Transcriptome analysis of miRNAs in subcutaneous adipose tissue of Duolang sheep, and Small Tail Han sheep was performed using RNA-Seq technology. Differentially expressed miRNAs were screened between different breeds. Target genes were predicted, and then the joint analysis of candidate genes were conducted based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. Finally, the RNA-Seq data were verified by real-time quantitative polymerase chain reaction (qRT-PCR). Herein, we identified 38 differentially expressed miRNAs (9 novel miRNAs and 29 known miRNAs). In addition, a total of 854 target genes were predicted by miRanda software. GO and KEGG pathway analysis demonstrated that regulation of lipolysis in adipocytes plays a key role in the deposition of subcutaneous adipose tissue in Duolang sheep and Small Tail Han sheep. The miRNAs might regulate fat deposits by regulating genes involved in regulation of lipolysis in adipocytes. Specifically, NC_ 040278.1_ 37602, oar-mir-493-3p, NC_ 040278.1_ 37521 and NC_ 040255.1_ 11627 might target PTGS2, AKT2, AKT3, and PIK3CA, respectively, and then play critical regulatory role. In conclusion, all the results provide a good idea for further revealing the mechanism of subcutaneous adipose tissue deposition and improving the meat production performance of sheep, and lay a foundation for promoting the development of animal husbandry.

[Screening of high-yield PUFAs Mortierella isabellina strain].

The original strain Mortierella isabellina As3.3410 was treated by microwave and ultraviolet. Mutated strains were screened by acetyl salicylic acid and low temperature (15°C). A high-yield strain named as A35-4 was successfully selected. The biomass of this strain was 17.9 g/L, oil content was 67.8%, oil production was 12.12 g/L, polyunsaturated fatty acids (PUFAs) content was 20.2%, and production of PUFAs was 2.46 g/L, which increased 32.6%, 49.8%, 98.69%, 14.0%, and 125.7% compared with the original A0 stain, respectively. The continuous slope transmission experiments confirmed that the strain had a good genetic stability. The study is beneficial for cloning high efficiency genes for PUFAs and producing PUFAs in this stain, and lays the ground work for creation of transgenic plants containing high levels of PUFAs.

Biogeochemical transfer and dynamics of iodine in a soil-plant system.

Radioactive iodide (125I) is used as a tracer to investigate the fate and transport of iodine in soil under various leaching conditions as well as the dynamic transfer in a soil-plant (Chinese cabbage) system. Results show that both soils (the paddy soil and the sandy soil) exhibit strong retention capability, with the paddy soil being slightly stronger. Most iodine is retained by soils, especially in the top 10 cm, and the highest concentration occurs at the top most section of the soil columns. Leaching with 1-2 pore volume water does not change this pattern of vertical distributions. Early breakthrough and long tailing are two features observed in the leaching experiments. Because of the relatively low peak concentration, the early breakthrough is really not an environmental concern of contamination to groundwater. The long tailing implies that the retained iodine is undergoing slow but steady release and the soils can provide a low but stable level of mobile iodine after a short period. The enrichment factors of 125I in different plant tissues are ranked as: root > stem > petiole > leaf, and the 125I distribution in the young leaves is obviously higher than that in the old ones. The concentrations of 125I in soil and Chinese cabbage can be simulated with a dual-chamber model very well. The biogeochemical behaviors of iodine in the soil-cabbage system show that cultivating iodized cabbage is an environmentally friendly and effective technique to eliminate iodine deficiency disorders (IDD). Planting vegetables such as cabbage on the 129I-contaminated soil could be a good remediation technique worthy of consideration.

Graphene oxide and ABA cotreatment regulates root growth of Brassica napus L. by regulating IAA/ABA.

Previous studies have proven that graphene oxide (GO) regulates abscisic acid (ABA) and indole-3-acetic acid (IAA) contents and modulates plant root growth. To better understand the mechanism of plant growth and development regulated by GO and crosstalk between ABA and GO, Zhongshuang No. 9 seedlings were treated with GO and ABA. The results indicated that GO and ABA significantly affected the morphological properties and endogenous phytohormone contents in seedlings, and there was significant crosstalk between GO and ABA. ABA treatments combined with GO led to a rapid decrease in triphenyltetrazolium chloride (TTC) reduction intensity, and the inhibitory effect was enhanced with increasing ABA concentration. The treatments significantly affected the transcriptional levels of some key genes involved in the ABA, IAA, cytokinin (CTK), salicylic acid (SA), and ethane (ETH) pathways and increased the ABA and gibberellin (GA) contents in rapeseed seedlings. The effects of the treatments on the IAA and CTK contents were complex, but, importantly, the treatments suppressed root elongation. Correlation analysis also indicated that the relationship between root length and IAA/ABA could be described by a polynomial function: y = 88.11x2 - 25.15x + 4.813(R²â€¯= 0.912). The treatments increased the ACS2 transcript abundance for ETH biosynthesis and the ICS1 transcriptional level of the key genes involved in salicylic acid (SA) biosynthesis, as well as the downstream signaling genes CBP60 and SARD1. This finding indicated that ABA is an important factor regulating the effects of GO on the growth and development of Brassica napus L., and that ETH and SA pathways may be potential pathways involved in the response of rape seedlings to GO treatment.

Mechanism of iodine uptake by cabbage: effects of iodine species and where it is stored.

Iodine-enhanced vegetable has been proven to be an effective way to reduce iodine deficiency disorders in many regions. However, the knowledge about what mechanisms control plant uptake of iodine and where iodine is stored in plants is still very limited. A series of controlled experiments, including solution culture, pot planting, and field experiments were carried out to investigate the uptake mechanism of iodine in different forms. A new methodology for observing the iodine distribution within the plant tissues, based on AgI precipitation reaction and transmission electron microscope techniques, has been developed and successfully applied to Chinese cabbage. Results show that iodine uptake by Chinese cabbage was more effective when iodine was in the form of IO(3) (-) than in the form of I(-) if the concentration was low (<0.5 mg L(-1)), but the trend was opposite if iodine concentration was 0.5 mg L(-1) or higher. The uptake was more sensitive to metabolism inhibitor in lower concentration of iodine, which implies that the uptake mechanism transits from active to passive as the iodine concentration increases, especially when the iodine is in the form of IO(3) (-). The inorganic iodine fertilizer provided a quicker supply for plant uptake, but the higher level of iodine was toxic to plant growth. The organic iodine fertilizer (seaweed composite) provided a more sustainable iodine supply for plants. Most of the iodine uptake by the cabbage is intercepted and stored in the fibrins in the root while the iodine that is transported to the above-ground portion (shoots and leaves) is selectively stored in the chloroplasts.

Cloning and molecular characterization of a functional flavonoid 3'-hydroxylase gene from Brassica napus.

A flavonoid 3'-hydroxylase (F3'H) gene, denoted BnF3'H-1, was cloned from oilseed rape (Brassica napus). The gene of 3038 base pairs (bp) contains 3 introns. The complementary DNA (cDNA) consists of 1820bp and has an open reading frame of 1536bp encoding a polypeptide of 511 amino acids with a molecular weight of 56.62kDa and an isoelectric point of 7.08. BnF3'H-1 shows high homology to known F3'H genes, especially F3'H from Arabidopsis thaliana. Untranslated regions (UTRs) may play important roles in regulating the expression of BnF3'H-1. Besides containing a Kozak sequence, the first 77-bp region is C-rich but G-poor, and the 26-bp 5'-UTR contains 3 sites of ACCACT-like sequences. Alternative polyadenylation in the 3'-UTR is adopted by this gene to generate heterogeneous transcripts. Conserved domain search and motif characterization identified BnF3'H-1 as a cytochrome P450. All F3'H-featured motifs, VVVAAS, GGEK and VDVKG, are unchanged in BnF3'H-1. The N-terminal signal peptide/anchor and 3 transmembrane helices were predicted in BnF3'H-1, and its subcellular localization is most probably at the endoplasmic reticulum. Since 16 phosphorylation sites could be predicted, phosphorylation may be a necessary post-translational modification of BnF3'H-1. The secondary structure is dominated by alpha-helices and random coils. Most helices are located in the middle region, while extended strands mainly intersperse in terminal regions. DNA gel blot analysis indicated that 2 different F3'H genes might exist in B. napus. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and RNA gel blot analysis showed that flowers have the highest F3'H expression, followed by pericarp and seed, and lower levels in some other organs. This species-featured expression pattern is in obedience to multiple functional roles that F3'H gene(s) play(s) in various organs of B. napus. The BnF3'H-1 coding region was expressed in Escherichia coli, and enzyme activity of the His-tagged protein was demonstrated by monitoring the conversion of the substrate naringenin using high-performance liquid chromatography (HPLC), suggesting that BnF3'H-1 is catalytically functional. RT-PCR analysis suggests that transcription level of the F3'H gene(s) is not the reason for the different seed colorations found in near-isogenic lines (black-seeded L1 and yellow-seeded L2) of B. napus.

Graphene oxide modulates root growth of Brassica napus L. and regulates ABA and IAA concentration.

Researchers have proven that nanomaterials have a significant effect on plant growth and development. To better understand the effects of nanomaterials on plants, Zhongshuang 11 was treated with different concentrations of graphene oxide. The results indicated that 25-100mg/l graphene oxide treatment resulted in shorter seminal root length compared with the control samples. The fresh root weight decreased when treated with 50-100mg/l graphene oxide. The graphene oxide treatment had no significant effect on the Malondialdehyde (MDA) content. Treatment with 50mg/l graphene oxide increased the transcript abundance of genes involved in ABA biosynthesis (NCED, AAO, and ZEP) and some genes involved in IAA biosynthesis (ARF2, ARF8, IAA2, and IAA3), but inhibited the transcript levels of IAA4 and IAA7. The graphene oxide treatment also resulted in a higher ABA content, but a lower IAA content compared with the control samples. The results indicated that graphene oxide modulated the root growth of Brassica napus L. and affected ABA and IAA biosynthesis and concentration.

[Radial artery recurrent branch of styloid process bone flap transfer for treatment of scaphoid nonunion].

OBJECTIVE: To investigate the clinical efficacy of radial artery recurrent branch of the styloid process bone flap transfer for treatment of scaphoid nonunion. METHODS: From 2002 to 2007, 18 patients with scaphoid nonunion included 12 males and 16 females with an average age of 31.2 years old ranging from l8 to 51 years. Time from injury to operation was 8 to 26 months (averaged 12.5 months). Radial bone flap blood supply by radial artery recurrent branch of styloid process was embedded in retrograde scaphoid shift distance at both ends, Kirschner wire fixed on the bone flap and bone fractures, and the radial styloid process was resected. Cast gypsum immobilization in a neutral place of wrist were performed for 6 to 8 weeks, the rehabilitation of physical therapy and functional exercise were performed after removal. Kirschner wire were removed after X-ray showed fracture healing. RESULTS: Among them, 15 cases were followed-up for from 8 to 25 months (means 13 months). All fractures healed. The healing time was 8 to 12 weeks. According to adult forearm fracture evaluation of Anderson the results were excellent in 14 cases, good in 1. CONCLUSION: The radial artery recurrent branch of styloid process bone flap has a wealth of blood circulation, can make free grafting of the creeping substitution process directly into the healing process.

[Characteristics of iodine uptake and accumulation by vegetables].

With seaweed iodine and KI as exogenous iodine sources, a pot experiment was conducted to study the characteristics of iodine uptake and accumulation by pakchoi cabbage, celery, capsicum, and radish. The results showed that the iodine content in the edible parts of test vegetables increased with increasing amount of exogenous iodine, but the iodine accumulation rate differed with the kinds of vegetables, in the order of pakchoi > celery > radish > capsicum. The majority of iodine was accumulated in roots, with lesser amount transferred to shoots. The distribution of iodine in vegetables was commonly in the order of root > leaf > stem > fruit, but the iodine in radish is lower in its rhizome than in its shoot. Low concentrations (0-25 mg x kg(-1)) of exogenous iodine had little effects on the growth of vegetables, while high concentrations (> or = 50 mg x kg(-1)) of it had inhibitory effects, resulting in a decreased vegetable biomass. The sensitivity of test vegetables to the adverse effect of exogenous iodine was in the order of capsicum > pachoi > celery > radish. Compared with seaweed iodine, KI decreased the biomass of first cutting significantly (P < 0.05), but for the second cutting, little difference was observed between these two iodine sources. The uptake and accumulation of these two iodine sources by vegetables also differed with cuttings, i.e., the first cutting vegetables absorbed more KI, while the second cutting vegetables absorbed more seaweed iodine (P < 0.05), suggesting that seaweed iodine had a longer efficacy than KI.