miR-128-3p Regulates Follicular Granulosa Cell Proliferation and Apoptosis by Targeting the Growth Hormone Secretagogue Receptor.

The proliferation and apoptosis of granulosa cells (GCs) affect follicle development and reproductive disorders, with microRNAs playing a crucial regulatory role. Previous studies have shown the differential expression of miR-128-3p at different stages of goat follicle development, which suggests its potential regulatory role in follicle development. In this study, through the Cell Counting Kit-8 assay, the EDU assay, flow cytometry, quantitative real-time polymerase chain reaction, Western blot, and the dual-luciferase reporter assay, we used immortal human ovarian granulosa tumor cell line (KGN) cells as materials to investigate the effects of miR-128-3p and its predicted target gene growth hormone secretagogue receptor (GHSR) on GC proliferation and apoptosis. The results show that overexpression of miR-128-3p inhibited the proliferation of KGN cells, promoted cell apoptosis, and suppressed the expression of proliferating cell nuclear antigen (PCNA) and B-cell lymphoma-2 (BCL2) while promoting that of Bcl-2 associated X protein (BAX). The dual-luciferase reporter assay revealed that miR-128-3p bound to the 3' untranslated region sequence of GHSR, which resulted in the inhibited expression of GHSR protein. Investigation of the effects of GHSR on GC proliferation and apoptosis revealed that GHSR overexpression promoted the expression of PCNA and BCL2, enhanced GC proliferation, and inhibited cell apoptosis, whereas the opposite effects were observed when GHSR expression was inhibited. In addition, miR-128-3p and GHSR can influence the expression of extracellular signal-regulated kinase 1/2 protein. In conclusion, miR-128-3p inhibits KGN cell proliferation and promotes cell apoptosis by downregulating the expression of the GHSR gene.

Expression profile and bioinformatics analysis of circRNA and its associated ceRNA networks in longissimus dorsi from Lufeng cattle and Leiqiong cattle.

This paper aims to explore the role of circRNA expression profiles and circRNA-associated ceRNA networks in the regulation of myogenesis in the longissimus dorsi of cattle breeds surviving under subtropical conditions in southern China by RNA sequencing and bioinformatics analysis. It also aims to provide comprehensive understanding of the differences in muscle fibers in subtropical cattle breeds and to expand the knowledge of the molecular networks that regulate myogenesis. With regard to meat quality indicators, results showed that the longissimus dorsi of LQC had lower pH (P < 0.0001), lower redness (P < 0.01), lower shear force (P < 0.05), and higher brightness (P < 0.05) than the longissimus dorsi of LFC. With regard to muscle fiber characteristics, the longissimus dorsi of LQC had a smaller diameter (P < 0.0001) and higher density of muscle fibers (P < 0.05). The analysis results show that the function of many circRNA-targeted mRNAs was related to myogenesis and metabolic regulation. Furthermore, in the analysis of the function of circRNA source genes, we hypothesized that btacirc_00497 and btacirc_034497 may regulate the function and type of myofibrils by affecting the expression of MYH6, MYH7, and NEB through competitive linear splicing.

Integrating Analysis to Identify Differential circRNAs Involved in Goat Endometrial Receptivity.

Endometrial receptivity is one of the main factors underlying a successful pregnancy, with reports substantiating the fact that suboptimal endometrial receptivity accounts for two-thirds of early implantation event failures. The association between circRNAs and endometrial receptivity in the goat remains unclear. This study aims to identify potential circRNAs and regulatory mechanisms related to goat endometrial receptivity. Therefore, the endometrial samples on day 16 of pregnancy and day 16 of the estrous cycle were analyzed using high-throughput RNA-seq and bioinformatics. The results show that 4666 circRNAs were identified, including 7 downregulated and 11 upregulated differentially expressed circRNAs (DE-circRNAs). Back-splicing and RNase R resistance verified the identified circRNAs. We predicted the competing endogenous RNA (ceRNA) regulatory mechanism and potential target genes of DE-circRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of these predicted target genes suggest that DE-circRNAs were significantly involved in establishing endometrial receptivity. Furthermore, Sanger sequencing, qPCR, correlation analysis and Fluorescence in Situ Hybridization (FISH) show that circ_MYRF derived from the host gene myelin regulatory factor (MYRF) might regulate the expression of interferon stimulating gene 15 (ISG15), thereby promoting the formation of endometrial receptivity. These novel findings may contribute to a better understanding of the molecular mechanisms regulating endometrial receptivity and promoting the maternal recognition of pregnancy (MRP).

Polymorphisms in BMPR-IB gene and their association with litter size trait in Chinese Hu sheep.

Identification and utilization of sheep major fecundity genes offer opportunities for the increase in litter size, as well as the improvement of production efficiency in livestock industry. BMPR-IB gene belongs to the TGF-ß superfamily, and is also considered as a regulator for sheep reproductive performance due to its involvement in the mammalian gametogenesis pathway. This study aimed to detect the variations of BMPR-IB gene in Hu sheep (N = 934) and to evaluate their effects on the litter size trait. qRT-PCR results showed that the mRNA expression level of BMPR-IB in kidney was the highest. And in the tissues of ovary and pituitary, the expression levels of prolific group were significantly higher than that of non-prolific group (p < 0.05). Through DNA sequencing and PCR-RFLP, three SNPs were identified in the genomic region of BMPR-IB gene; the individuals with CC in g.29362047T > C, AA in g.29427689G > A and GG in FecB had better fecundity characterization. Additionally, association analysis indicated that two diplotypes of Hap2/2 and Hap2/4 showed larger litter size. Overall, our results verified several useful markers which would contribute to further development of sheep breeding strategies.

Genome-wide prediction and comparative transcriptomic analysis reveals the G protein-coupled receptors involved in gonadal development of Apostichopus japonicus.

The sea cucumber Apostichopus japonicus is dioecious, with seasonal reproduction. G protein-coupled receptor (GPCR)-mediated signaling systems might play critical roles in the reproductive control of A. japonicus. Here, we classified GPCR from the genome in silico and used transcriptomic analyses to further mine those that function in gonadal-development control. Totally, 487 GPCRs were predicted from A. japonicus, and 183 of these were further annotated to molecular pathways. Transcriptome analysis revealed 327 GPCRs expressed in gonads, and these were classified into four families and 19 subfamilies. Three pathways were apparently associated with reproduction, including neuroactive ligand-receptor interaction, the mTOR and Wnt signaling pathways. Seven and eight ovary- and testis-specific GPCRs were filtered, and the gene expression profiles were determined in multiple tissues and gonads at different developmental stages by qPCR. These results provide new insights into the discovery of GPCR-mediated signaling control in sea cucumber reproduction, especially in gonadal development control.

miR-450-5p and miR-202-5p Synergistically Regulate Follicle Development in Black Goat.

Follicle maturation is a complex biological process governed by numerous factors, and researchers have observed follicle development by studying the proliferation and apoptosis of follicular granulosa cells (GCs). However, the regulatory mechanisms of GCs proliferation and death during follicle development are largely unknown. To investigate the regulatory mechanisms of lncRNAs, mRNAs, and microRNAs, RNA sequencing (RNA-seq) and small RNA-seq were performed on large (>10 mm) and small follicles (<3 mm) of Leizhou black goat during estrus. We discovered two microRNAs, miR-450-5p and miR-202-5p, which can target GCs in goats and may be involved in follicle maturation, and the effects of miR-450-5p and miR-202-5p on ovarian granulosa cell lines were investigated (KGN). Using cell counting kit-8 (CCK-8) assays, 5-Ethynyl-2'-deoxyuridine (EdU) assay and flow cytometry, miR-202-5p overexpression could suppress the proliferation and induce apoptosis of GCs, whereas miR-450-5p overexpression induced the opposite effects. The dual-luciferase reporter assay confirmed that miR-450-5p could directly target the BMF gene (a BCL2 modifying factor), and miR-202-5p targeted the BCL2 gene. A considerable rise in phosphorylated Akt (p-AKT) protein was observed following the downregulation of BMF by miR-450-5p mimics. After BMF gene RNAi therapy, a notable elevation in p-AKT was detected. Mimics of miR-202-5p inhibited BCL2 protein expression, significantly decreasing p-AMPK protein expression. These results imply that during the follicular development in black goats, the miR-450-5p-BMF axis favored GC proliferation on a wide scale, while the miR-202-5p-BCL2 axis triggered GC apoptosis.

Identification of genetic markers associated with milk production traits in Chinese Holstein cattle based on post genome-wide association studies.

With the rapid development of dairy industry, the breeding process of dairy cows has been accelerated. In previous genome-wide association studies (GWAS), a large number of genetic markers have been reported which may contribute to the selection of Holstein populations with superior milk-producing traits, but they remain to be further verified before practical application. In this study, 90 single nucleotide polymorphisms (SNPs) were selected, which were reported to be significantly associated with five milk production traits, including 305-day milk yield (305MY), 305-day milk fat percent (305FC), 305-day milk protein percent (305PC), 305-day milk fat yield (305FY) and 305-day milk protein yield (305PY). Effective 305-day data and fresh DNA samples were obtained from 295 healthy cows with gestational age of 1-4. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) was used to perform precise genotyping of these loci, followed by site association and haplotype analysis. Results showed that 36 out of 90 loci were supported to be used as genetic markers. In particular, several novel and effective haplotypes were also presented. Overall, our results verified tens of useful markers and provided a basis for further development of breeding strategies.

Comprehensive analysis of mRNAs and miRNAs in the ovarian follicles of uniparous and multiple goats at estrus phase.

BACKGROUND: Fertility is an important economic trait in the production of meat goat, and follicular development plays an important role in fertility. Although many mRNAs and microRNAs (miRNAs) have been found to play critical roles in ovarian biological processes, the interaction between mRNAs and miRNAs in follicular development is not yet completely understood. In addition, less attention has been given to the study of single follicle (dominant or atretic follicle) in goats. This study aimed to identify mRNAs, miRNAs, and signaling pathways as well as their interaction networks in the ovarian follicles (large follicles and small follicles) of uniparous and multiple Chuanzhong black goats at estrus phase using RNA-sequencing (RNA-seq) technique. RESULTS: The results showed that there was a significant difference in the number of large follicles between uniparous and multiple goats (P < 0.05), but no difference in the number of small follicles was observed (P > 0.05). For the small follicles of uniparous and multiple goats at estrus phase, 289 differentially expressed mRNAs (DEmRNAs) and 16 DEmiRNAs were identified; and for the large follicles, 195 DEmRNAs and 7 DEmiRNAs were identified. The functional enrichment analysis showed that DE genes in small follicles were significantly enriched in ovarian steroidogenesis and steroid hormone biosynthesis, while in large follicles were significantly enriched in ABC transporters and steroid hormone biosynthesis. The results of quantitative real-time polymerase chain reaction were consistent with those of RNA-seq. Analysis of the mRNA-miRNA interaction network suggested that CD36 (miR-122, miR-200a, miR-141), TNFAIP6 (miR-141, miR-200a, miR-182), CYP11A1 (miR-122), SERPINA5 (miR-1, miR-206, miR-133a-3p, miR-133b), and PTGFR (miR-182, miR-122) might be related to fertility, but requires further research on follicular somatic cells. CONCLUSIONS: This study was used for the first time to reveal the DEmRNAs and DEmiRNAs as well as their interaction in the follicles of uniparous and multiple goats at estrus phase using RNA-seq technology. Our findings provide new clues to uncover the molecular mechanisms and signaling networks of goat reproduction that could be potentially used to increase ovulation rate and kidding rate in goat.

Design, synthesis and interaction of BRC4 analogous peptides with RAD51(241-260).

Breast cancer susceptibility gene 2 (BRCA2) is an important tumor suppressor, which is participated in repair of damaged DNA by its highly conserved BRC repeat motifs regulating RAD51 protein homologous recombination and thereby preventing cell carcinogenesis. In this study, the BRCA2(1524-1548)-RAD51(241-260) complex structure was obtained based on PDB bank data 1N0W, which provided the basis for site-specific mutation of BRCA2(1524-1548). The BRC4 and BRC4 analogous peptides were synthesized, and the interaction between BRC peptide and RAD51(241-260) was studied by fluorescence spectroscopy, circular dichroism spectroscopy and microscale thermophoresis (MST). The results of circular dichroism showed that the changes in secondary structures of RAD51(241-260) occurred after adding BRC4 analogous peptides, and the α-helix content increased significantly. Fluorescence spectral data demonstrated that the model of BRC peptide binding to RAD51(241-260) was static quenching, and the binding constants of BRC4, P1, P2, P4 with RAD51(241-260) were 1.647 × 10-4 L mol-1, 2.532 × 10-4 L mol-1, 3.161 × 10-4 L mol-1, 1.705 × 10-4 L mol-1, respectively. The results of MST indicated that P2 and RAD51(241-260) have better affinity for dissociation constant 44.286 µM. The strongest affinity between P2 and RAD51(241-260) indicated that the mutation of amino acid residue constituting BRC α-helix affects the structure and interaction of BRC peptide and RAD51(241-260).

Design of BRC analogous peptides based on the complex BRC8-RAD51 and the preliminary study on the peptide structures.

BRCA2 is an important tumor suppressor gene that plays a critical role in preserving the stability of cellular genetic information, participating in DNA repair by engaging in binding interactions with RAD51 proteins. However, the lack of structural data on BRCA2 and RAD51 makes the study of their interaction mechanism still a great challenge. We characterize the structure of the BRC8-RAD51 complex using ZDOCK protein docking software and identify the potential non-conserved active site of BRC8 via virtual alanine scanning, utilizing the obtained results to synthesize BRC8, its six analogous peptides (BRC8-1 to BRC8-6), and critical peptide fragment of RAD51 (RAD51(231-260)) by Fmoc solid-phase synthesis. The analogous peptides are found to exhibit a secondary structure significantly different from that of BRC8 by circular dichroism spectroscopy, which indicates that mutation sites determined by computer-aided simulation correspond to key amino acid residues substantially affecting polypeptide structure. On the other hand, the secondary structure of RAD51(231-260) was also considerably influenced by its interaction with BRC8 and analogs, e.g., the fraction of the α-helical structure in RAD51(231-260) increased to 23.6, 15.1, and 13.5% upon interaction with BRC8-1, BRC8-3, and BRC8-6, respectively. The results show that the properties of C-terminal amino acid residues significantly influence peptide-peptide interactions, in agreement with the results of virtual alanine scanning. Therefore, computer-aided simulation was confirmed to be a technique that is useful for narrowing down the range of sites responsible for interactions between peptides or proteins, and provides new inspirations for the design of peptides with strong interactions.

Role of mRNAs and long non-coding RNAs in regulating the litter size trait in Chuanzhong black goats.

Fecundity improvement is one of the most important objectives for goat breeders as it can considerably greatly increase production efficiency. The molecular mechanisms underlying fecundity in goats remain largely unknown. To explore the molecular and genetic mechanisms related to the fecundities and prolificacies in Chuanzhong black goats, we performed high-throughput RNA sequencing to identify differentially expressed long non-coding RNAs (lncRNAs) and mRNAs (DElncRNAs and DEmRNAs, respectively) the ovaries of high-fecundity and low-fecundity goats; furthermore, we conducted functional annotation analyses to identify pathways of interest. Overall, 1,353 DEmRNAs and 168 DElncRNAs were identified. Quantitative real-time PCR (qRT-PCR) was performed to validate some randomly selected DElncRNAs and DEmRNAs. We found that two DElncRNAs ENSCHIT00000005909 and ENSCHIT00000005910 might positively influence the expression of the corresponding gene IL1R2 (upregulated in high-fecundity group), exerting co-regulative effects on the ovarian function, through which litter size might show variations. KEGG pathway analysis indicated that the DEmRNAs SRD5A2, LOC102191297 and LOC102171967 were significantly enriched in steroid hormone biosynthesis-this pathway was related to animal reproduction. To summarize, our findings expand the understanding pertaining to the biological functions of lncRNAs and contribute to the annotation of the goat genome; moreover, they should be helpful for further studying the role of lncRNAs in ovulation and lambing.

Lycopene supplementation regulates the gene expression profile and fat metabolism of breeding hens.

This study investigated the effects of lycopene on the gene expression profile and expression of genes related to fat metabolism of Xinghua breeding hens. Seven hundred and twenty healthy breeding hens were randomly assigned to four treatments; each treatment was replicated six times with 30 hens each. Broken rice and soybean meal were adopted for the basal diet and added with 0 (control group), 20, 40 and 80 mg/kg lycopene respectively. Gene expression profile of the liver induced by lycopene and expression of genes related to fat metabolism in hens liver and intestine were analysed after 42-day feeding trial including 7-day pre-feeding period and 35-day formal period. The genes involved in fat metabolism were analysed, and we found that lycopene significantly increased the expression of PGC1α, PPARα, RXRα and RARα in the liver, PPARγ, RXRα and RXRγ in the jejunum, and RARα in the duodenum (p < .05); reduced the expression of FABP1 and FABP10 in the liver, and FATP4 in the jejunum (p < .05). By analysing gene expression profile, 158 differentially expressed genes (DEGs) including 69 up-regulated genes and 89 down-regulated genes were obtained between control group and 40 mg/kg group. KEGG pathway analysis was performed on all DEGs, and 5 pathways were obtained. In conclusion, lycopene can affect the expression of related genes, and this may be one of the reasons that lycopene can regulate fat metabolism.

High-performance voltammetric sensor for dichlorophenol based on ß-cyclodextrin functionalized boron-doped graphene composite aerogels.

2, 2-methylenebis (4-chlorophenol) (dichlorophenol, Dcp) is a priority pollutant that poses a serious health threat to the public. Thus, the sensitive analysis of Dcp is of great significance. Heteroatom-doped carbon nanomaterials modified electrodes have been proven to be good electrocatalysts for electrochemical sensing application. ß-cyclodextrin (ß-CD) as a signal amplifier has also been utilized in biosensors. Inspired by these, in this study, a new composite of ß-CD and three-dimensional (3D) boron-doped graphene aerogels (BGAs/ß-CD) has been designed as a high-performance electrochemical sensing platform for Dcp determination. Graphene aerogels possess high specific surface area, large pore volume and good conductivity, which ensure rapid mass transfer and accelerated electron transfer. Besides, boron doping causes uneven charge distribution on the graphene lattice surface, producing a large amount of flowing π electrons, which provide abundant active sites for the catalytic oxidation reaction of Dcp. In addition, Dcp molecules could be captured into ß-CD through host-guest recognition, which can effectively amplify the detection signal. Combining the merits of BGAs and ß-CD, the BGAs/ß-CD based sensor achieved sensitive detection of Dcp. Under optimized experimental conditions, the oxidation currents and the concentration of Dcp had a good linear relationship within 1.0 nM âˆ¼ 21 µM. The detection limit was estimated as 0.33 nM (S/N = 3). This study might provide a new basis for the fabrication of 3D BG-based aerogel architectural material and its application in Dcp detection.

Ratiometric electrochemical sensor for sensitive detection of sunset yellow based on three-dimensional polyethyleneimine functionalized reduced graphene oxide aerogels@Au nanoparticles/SH-ß-cyclodextrin.

Electrochemical methods have been deemed effective strategies for the detection of dye additive sunset yellow (SY) owing to their low cost, good stability, and high sensitivity. However, the application of the existing sensors with single electrical signal response is limited by their inadequate sensitivity and large background interference. Herein, a ratiometric electrochemical strategy with a dual signal was developed to detect SY. The strategy had an intrinsic built-in correction to the effects from the system, and thus reduced the influence of environmental change. 3D polyethyleneimine functionalized reduced graphene oxide aerogels@Au nanoparticles/SH-ß-cyclodextrin (PEI-rGAs@AuNPs/SH-ß-CD) was used as the sensing material due to its 3D macroporous microstructure with high specific surface area and excellent electronic conductivity. Guest molecule methylene blue (MB) was chosen as a probe molecule, which formed an inclusion host-guest complex with a SH-ß-CD host in advance. The target molecule SY displaced MB from the CD cavities, resulting in the decrease of MB current and the increase of SY current. With the logarithmic value of ISY/IMB as the readout signal, the detection limit of the developed ratiometric electrochemical sensor reached as low as 0.3 nM, confirming the excellent sensitivity. Furthermore, this strategy exhibited good selectivity and repeatability, and could be used for the detection of SY in a real sample.

A CuO-CeO2 composite prepared by calcination of a bimetallic metal-organic framework for use in an enzyme-free electrochemical inhibition assay for malathion.

An enzyme-free electrochemical method is described for the determination of trace levels of malathion. It is based on a nanostructured copper-cerium oxide (CuO-CeO2) composite prepared by calcination of a Cu(II)/Ce(III) metal-organic framework. The morphology, crystal structure and elemental composition of composite was studied by scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction and X-ray photoelectron spectroscopy. The principle for malathion determination is based on the fact that the redox signal of CuO (best measured at around -0.1 V vs. SCE) (at 100 mV/s) is inhibited by malathion due to affinity between CuO and the sulfur groups of malathion. The introduction of CeO2 into the composite system further improves the analytical performance. This is attributed to the unique microstructure and the synergistic effect between CuO and CeO2. Experimental parameters like solution pH value, Cu/Ce molar ratio, accumulation potential, accumulation time, and CuO-CeO2 volume on the electrode were optimized. The assay has a linear range of 10 fM to 100 nM and a 3.3 fM detection limit (at S/N = 3). The electrode is selectively inhibited by malathion even in the presence of potentially interfering substances. Graphical abstract A sensitive and effective enzyme-free electrochemical sensor has been developed for the detection of malathion based on CuO-CeO2 composite derived from bimetallic metal-organic frameworks.

Sensitive determination of nitrite by using an electrode modified with hierarchical three-dimensional tungsten disulfide and reduced graphene oxide aerogel.

Nanosheets of tungsten disulfide (WS2) were used to improve the physicochemical properties of reduced graphene oxide aerogel (rGA). The nanosheets were directly integrated into 3D hybrid architecture of rGA by a solvothermal mixing method by which the WS2 sheets were assembled onto the conductive graphene network. WS2 with highly exfoliated and defect-rich structure made the WS2/rGA composite possess plentiful active sites, and this enhanced the electrocatalytic capability of the composite. The introduction of poorly conductive WS2 into 3D rGA system decreases the background current of rGA when used as electrode material. This is advantageous in terms of signal to-noise ratio and analytical performance in general. The WS2/rGA electrode, best operated at a potential of 0.68 V (vs. SCE) has a linear response in the 0.01 to 130 µM nitrite concentration range with a low detection limit of 3 nM (at S/N = 3). It is selective, reproducible, stable and is successfully applied to the determination of nitrite in spiked bacon samples. Graphical Abstract Schematic presentation of an electrochemically modified electrode for the detection of nitrite based on 3D tungsten disulfide/reduced graphene oxide aerogel (WS2/rGA).

Evaluation of pigeon pea leaves (Cajanus cajan) replacing alfalfa meal on growth performance, carcass trait, nutrient digestibility, antioxidant capacity and biochemical parameters of rabbits.

A 30-day experiment was performed to determine the effect of pigeon pea leaves (PPL) on growth performance, carcass trait, meat quality, nutrient digestibility, antioxidant capacity and biochemical parameters of growing rabbits. In a completely randomized design, PPL replaced alfalfa meal at the level of 0%, 10%, 20% and 30%, which were named PPL0 (control), PPL10, PPL20 and PML30 respectively. Two hundred New Zealand white rabbits at 6 weeks with similar weight (870.23 ± 15.98 g) were allocated to four dietary groups with five replicates containing 10 rabbits/per replicate (male). The results showed that: (a) PPL powder contained 24.26% crude protein, 4.34% crude fat, 17.86% crude fibre, 7.05% ash, 1.35% calcium, 0.28% phosphorus, 1.09% lysine and 0.20% methionine, and the chemical compositions are on DM basis; (b) the ratio of feed to gain of rabbits fed diet PPL10 was significantly better (p < 0.05) than those fed other three diets; (c) the content of longissimus dorsi (LD) moisture in the rabbits fed diets without PPL (control group) was 12% lower than that in the PPL30 diets (60.1 vs. 72.1; p < 0.05). In PPL10, PPL20 and PPL30 diets, the leg muscle (LM) b*(yellowness) value was 33%, 30% and 22.6% higher than the control group respectively. The rabbits fed diets PPL0 had lower (p < 0.05) LM crude protein and ash and higher (p < 0.05) crude fat of LD and LM as compared with those fed other diets; (d) crude protein and energy digestibility of PPL0 and PPL10 diets were significantly higher (p < 0.05) than PPL30 diets; and (e) serum glutathione peroxidase (GSH-Px) activity of the rabbits fed PPL10 and PPL30 diets was significantly higher (p < 0.05) than that fed PPL20 diets. Liver total antioxidant capacity (T-AOC) activity of the PPL30 groups was 1.3% higher (p < 0.05) than the PPL10 group. Additionally, the control group (PPL0) had the highest (p < 0.05) blood urea nitrogen (BUN), total cholesterol (TCHO) and low-density lipoprotein cholesterol (LDLC) content compared with the groups supplemented with PPL. The PPL30 group had the highest (p < 0.05) triiodothyronine (T3 ) and tetraiodothyroxine (T4 ) value among the dietary groups.

The role of regulator FucP in Edwardsiella tarda pathogenesis and the inflammatory cytokine response in tilapia.

The animal intestine provides a competitive environment for the microbiota. Successful colonization by pathogens requires sensing chemical signals to regulate the expression of virulence genes. Some bacteria rely on a two-component chemical signal transduction system, named FusKR, to regulate virulence genes expression by intestinal fucose. Here we construct the fucP gene deletion strain prove FucP regulation of the T3SS in E. tarda. The result showed that the mutant strain had down-regulated significantly the gene expression of FusKR and T3SS compared to the wild-type strain (P < 0.05). This mutant strain significantly increased LD50 in zebrafish compared to the wild-type strain (P < 0.05), and significantly decreased penetration and motility in mucin than the wild-type strain (P < 0.05). Meanwhile, tilapia infected with mutant strain show significantly reduced E. tarda adherence and colonization than those infected with the wild-type strain (P < 0.05). Fish infected with EIB202 and ΔfucP showed significantly higher (P < 0.05) gene expression of IL-1ß, TNF-α, IFN-γ, TGF-ß and HSP-70 in head kidney than fish treated with PBS in the whole observed period; however CPP-3 did not show significant differences (P > 0.05) in all groups. Fish infected with EIB202 showed significantly higher (P < 0.05) gene expression of TGF-ß in head kidney than fish treated with ΔfucP in the whole observed period; however other cytokines did not show significant differences (P > 0.05) in the whole observed period. In addition, Fish infected with EIB202 showed significantly higher (P < 0.05) gene expression of IL-1ß, TNF-α and TGF-ß in spleen than fish treated with ΔfucP in the whole observed period, however IFN-γ, CPP-3, and HSP-70 did not show significant differences (P > 0.05) in the whole observed period. Although the gene expression of cytokines was induced similarly by both strains, all results indicate that the fucP gene deletion down-regulates the key gene expression of FucKR and T3SS, reduces the pathogenicity of E. tarda in fish, particularly decreases inducing the gene expression of TGF-ß in the head kidney and IL-1ß, TNF-α and TGF-ß in the spleen.

Expression profiling reveals genes involved in the regulation of wool follicle bulb regression and regeneration in sheep.

Wool is an important material in textile manufacturing. In order to investigate the intrinsic factors that regulate wool follicle cycling and wool fiber properties, Illumina sequencing was performed on wool follicle bulb samples from the middle anagen, catagen and late telogen/early anagen phases. In total, 13,898 genes were identified. KRTs and KRTAPs are the most highly expressed gene families in wool follicle bulb. In addition, 438 and 203 genes were identified to be differentially expressed in wool follicle bulb samples from the middle anagen phase compared to the catagen phase and the samples from the catagen phase compared to the late telogen/early anagen phase, respectively. Finally, our data revealed that two groups of genes presenting distinct expression patterns during the phase transformation may have important roles for wool follicle bulb regression and regeneration. In conclusion, our results demonstrated the gene expression patterns in the wool follicle bulb and add new data towards an understanding of the mechanisms involved in wool fiber growth in sheep.

Effect of salinity on the toxicokinetics, oxidative stress, and metallothionein gene expression in Meretrix meretrix exposed to cadmium.

To understand the effect of salinity on the toxicokinetics, oxidative stress, and detoxification of cadmium-exposed Meretrix meretrix, M. meretrix were acclimatized to different salinities (8, 14, 20, 26, and 32 ppt) for 14 d, exposed to 10 µg/L Cd for 7 d, followed by a 28-day depuration period. The internal Cd concentration was determined, and the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST)), and the malondialdehyde (MDA) content were measured. The mRNA expression levels of antioxidant enzyme (Cu/Zn SOD, CAT) and detoxification-related genes metallothionein (MT) were analyzed. The mean concentrations of Cd in M. meretrix tissues were in the order gill > digestive gland > mantle > axe foot. The Cd uptake rate in the four tissues decreased with increasing salinity (range: 14-26 ppt). The Cd elimination half-lives were the highest at 8 ppt and 14 ppt salinity. Cadmium activated the four oxidative stress-related related enzymes in the gills. At the end of accumulation period, Cd exposure at 20 ppt salinity significantly increased the expression of Cu/Zn SOD. CAT expression was significantly inhibited at 20 ppt salinity, but was induced at 32 ppt. MT mRNA expression was only induced under Cd at 20 ppt salinity. At the end of depuration period, Cu/Zn SOD expression was inhibited at salinities of 8, 14, and 26 ppt. The results indicated that SOD, CAT, GST, MDA, Cu/Zn SOD, CAT, and MT were sensitive to cadmium in a water environment, and can be used as indicators of marine heavy metal pollution.