Whole genome sequencing identified genomic diversity and candidated genes associated with economic traits in Northeasern Merino in China.

Introduction: Northeast Merino (NMS) is a breed developed in Northeast China during the 1960s for wool and meat production. It exhibits excellent traits such as high wool yield, superior meat quality, rapid growth rate, robust disease resistance, and adaptability to cold climates. However, no studies have used whole-genome sequencing data to investigate the superior traits of NMS. Methods: In this study, we investigated the population structure, genetic diversity, and selection signals of NMS using whole-genome sequencing data from 20 individuals. Two methods (integrated haplotype score and composite likelihood ratio) were used for selection signal analysis, and the Fixation Index was used to explore the selection signals of NMS and the other two breeds, Mongolian sheep and South African meat Merino. Results: The results showed that NMS had low inbreeding levels, high genomic diversity, and a pedigree of both Merino breeds and Chinese local breeds. A total length of 14.09 Mb genomic region containing 287 genes was detected using the two methods. Further exploration of the functions of these genes revealed that they are mainly concentrated in wool production performance (IRF2BP2, MAP3K7, and WNT3), meat production performance (NDUFA9, SETBP1, ZBTB38, and FTO), cold resistance (DNAJC13, LPGAT1, and PRDM16), and immune response (PRDM2, GALNT8, and HCAR2). The selection signals of NMS and the other two breeds annotated 87 and 23 genes, respectively. These genes were also mainly focused on wool and meat production performance. Conclusion: These results provide a basis for further breeding improvement, comprehensive use of this breed, and a reference for research on other breeds.

Correction: Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus.

[This corrects the article DOI: 10.1371/journal.pone.0270708.].

Regulatory Effects of FGF9 on Dermal Papilla Cell Proliferation in Small-Tailed Han Sheep.

Fibroblast growth factor 9 (FGF9) is crucial for the growth and development of hair follicles (HFs); however, its role in sheep wool growth is unknown. Here, we clarified the role of FGF9 in HF growth in the small-tailed Han sheep by quantifying FGF9 expression in skin tissue sections collected at different periods. Moreover, we evaluated the effects of FGF9 protein supplementation on hair shaft growth in vitro and FGF9 knockdown on cultured dermal papilla cells (DPCs). The relationship between FGF9 and the Wnt/ß-catenin signaling pathway was examined, and the underlying mechanisms of FGF9-mediated DPC proliferation were investigated. The results show that FGF9 expression varies throughout the HF cycle and participates in wool growth. The proliferation rate and cell cycle of FGF9-treated DPCs substantially increase compared to that of the control group, and the mRNA and protein expression of CTNNB1, a Wnt/ß-catenin signaling pathway marker gene, is considerably lower than that in the control group. The opposite occurs in FGF9-knockdown DPCs. Moreover, other signaling pathways are enriched in the FGF9-treated group. In conclusion, FGF9 accelerates the proliferation and cell cycle of DPCs and may regulate HF growth and development through the Wnt/ß-catenin signaling pathway.

Reverse transcription recombinase-aided amplification assay for avian influenza virus.

Avian influenza virus (AIV) infection can lead to severe economic losses in the poultry industry and causes a serious risk for humans. A rapid and simple test for suspected viral infection cases is crucial. In this study, a reverse transcription recombinase-aided amplification assay (RT-RAA) for the rapid detection of all AIV subtypes was developed. The reaction temperature of the assays is at 39 °C and the detection process can be completed in less than 20 min. The specificity results of the assay showed that this method had no cross-reaction with other main respiratory viruses that affect birds, including Newcastle disease virus (NDV) and infectious bronchitis virus (IBV). The analytical sensitivity at a 95% confidence interval was 102 RNA copies per reaction. In comparison with a published assay for reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), the κ value of the RT-RAA assay in 384 avian clinical samples was 0.942 (p < 0.001). The sensitivity and specificity of the RT-RAA assay for avian clinical sample detection was determined as 97.59% (95% CI 93.55-99.23%) and 96.79% (95% CI 93.22-98.59%), respectively. The RT-RAA assay for AIV in this study provided an effective and practicable tool for AIV molecular detection.

Molecular epidemiology analysis of fowl adenovirus detected from apparently healthy birds in eastern China.

BACKGROUND: Fowl adenovirus is of major concern to the poultry industry worldwidely. In order to monitor the prevalent status of Fowl adenovirus in China, a total of 1920 clinical samples from apparently healthy birds in the 25 sites of poultry flocks, Slaughterhouse and living bird markets from 8 provinces in eastern China were collected and detected by PCR, sequencing, and phylogenetic analysis. RESULTS: The epidemiological survey showed that Fowl adenoviruses were detected in living bird markets, and circulating in a variety of fowl species, including chickens, ducks, goose and pigeons. Among the 1920 clinical samples, 166 samples (8.65%) were positive in the fowl adenovirus PCR detection. In this study, totally all the 12 serotypes (serotypes of 1, 2, 3, 4, 5, 6, 7, 8A, 8B, 9, 10 and 11) fowl adenoviruses were detected, the most prevalent serotype was serotype 1. Phylogenetic analysis indicated that 166 FAdVs of 12 serotypes were divided into 5 fowl adenovirus species (Fowl aviadenovirus A, B, C, D, E). CONCLUSIONS: In the epidemiological survey, 8.65% of the clinical samples from apparently healthy birds were positive in the fowl adenovirus PCR detection. Totally all the 12 serotypes fowl adenoviruses were detected in a variety of fowl species, which provided abundant resources for the research of fowl adenoviruses in China. The newly prevalent FAdV serotypes provides valuable information for the development of an effective control strategy for FAdV infections in fowls.

Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus.

In order to develop an appropriate method for high-throughput detection of avian metapneumovirus, a quadruple real-time reverse-transcription polymerase chain reaction assay was established with four pairs of specific primers and four specific probes based on the G or M gene of aMPV-A, aMPV-B, aMPV-C and aMPV-D. Its specificity and sensitivity were evaluated, and clinical samples were tested by the method. The results showed that all the four subgroups of avian metapneumovirus can be detected in the quadruple real-time RT-PCR assay simultaneously, with a detection limit of 100-1000 cRNA copies/reaction. The other common poultry viruses were negative. In the avian clinical sample detection, 39 out of 1920 clinical samples collected from 8 provinces were positive. Compared with published RT-PCR assays, the κ value of the quadruple real-time RT-PCR assay in 1920 avian clinical samples was 1.000 (P < 0.001). The established method could be used for the rapid detection of the four subgroups of avian metapneumovirus with high specificity and high sensitivity.

Immunomodulatory effects of Inonotus obliquus polysaccharide on splenic lymphocytes infected with Toxoplasma gondii via NF-κB and MAPKs pathways.

CONTEXT: As a medicinal and edible fungus, Inonotus obliquus has been traditionally used to prevent and treat various ailments. Inonotus obliquus polysaccharide (IOP) isolated from I. obliquus processes many biological activities, our series of in vivo studies have shown that IOP protects against Toxoplasma gondii infection. OBJECTIVE: This study aimed to investigate the in vitro immunomodulatory effects and its mechanisms of IOP on mouse splenic lymphocytes infected with T. gondii. MATERIALS AND METHODS: Mouse splenic lymphocytes were infected with T. gondii tachyzoites, and treated with different concentrations of IOP. The levels of cytokines and chemokines were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The expression of toll-like receptor 2 (TLR2) and TLR4, and the modulation of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) signaling pathways were determined by Western blot. RESULTS: IOP significantly decreased the over-release of cytokine interleukin-1 beta (IL-1ß), IL-4, IL-6, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) in supernatant from T. gondii-infected splenic lymphocytes. IOP also effectively inhibited the overexpression of cytokines and chemokine macrophage inflammatory protein-1 (MIP-1) and monocyte chemoattractant protein-1 (MCP-1) mRNA. Furthermore, IOP down-regulated TLR2 and TLR4 expressions and inhibited the over-phosphorylation of NF-κB p65 and inhibitor κBα (IκBα) in NF-κB signaling pathway and p38, c-Jun N-terminal kinase (JNK) in MAPKs signaling pathway. By observing the effect of IOP on TNF-α secretion after pretreatment with specific inhibitors, it was further confirmed that IOP was involved in the regulation of NF-κB, p38, and JNK signaling pathways. CONCLUSIONS: These data indicate that IOP can inhibit the excessive inflammatory response caused by T. gondii infection through modulating NF-κB, p38, and JNK signaling pathways, and thus plays the in vitro anti-T. gondii role.

Curcumin Activates ROS Signaling to Promote Pyroptosis in Hepatocellular Carcinoma HepG2 Cells.

BACKGROUND/AIM: Curcumin is a polyphenol that exerts a variety of pharmacological activities and plays an anti-cancer role in many cancer cells. It was recently reported that gasdermin E (GSDME) is involved in the progression of pyroptosis. MATERIALS AND METHODS: HepG2 cells were treated with various concentrations of curcumin and cell viability was examined using MTT assay, apoptosis was analysed using flow cytometry, reactive oxygen species (ROS) levels using dihydroethidium, LDH release using an LDH cytotoxicity assay, and protein expression using western blot. RESULTS: Curcumin increased the expression of the GSDME N-terminus and proteins involved in pyrolysis, promoted HspG2 cell pyrolysis and increased intracellular ROS levels. Moreover, inhibition of the production of intracellular ROS with n-acetylcysteine (NAC) improved the degree of apoptosis and pyrolysis induced by curcumin. CONCLUSION: Curcumin induces HspG2 cell death by increasing apoptosis and pyroptosis, and ROS play a key role in this process. This study improves our understanding of the potential anti-cancer properties of curcumin in liver cancer.

Effects of E2 binding enzyme UBC9 on porcine oocyte maturation, apoptosis and embryo development.

SUMOylation is a dynamic post-translational modification process. However, the function of small ubiquitin-like modifiers (SUMOs) in the maturation of porcine oocytes and embryo growth is not well known. Therefore, the aim of this study was to investigate the effect of E2 binding enzyme UBC9 on the expression of SUMO-1 protein during the in vitro maturation of porcine oocytes and embryo development after in vitro fertilization. Four groups were used: 0 (Control), 5, 10 and 15 µg/ml UBC9. Western blotting, flow cytometry and RT-qPCR were used to detect the in vitro maturation of porcine oocytes, SUMO-1 content, viability and the expression of apoptotic genes. Compared to those in the control treatment, the maturation rate (p < .05) and viability (p < .01) of oocytes in the 5 µg/ml treatment group decreased significantly. SUMO-1 protein markers appeared at 59 and 71 kDa and the content of SUMO-1 protein in the 10 µg/ml treatment group decreased significantly (p < .05). In the expression of apoptosis-related genes, Bcl-2 gene expression was significantly downregulated in the 10 µg/ml treatment group (p < .05). However, Bax and Caspase-3 were significantly upregulated in the 5 µg/ml treatment group (p < .05). During embryonic development, the cleavage rate of oocytes in the 10 µg/ml treatment group was significantly reduced (p < .05), whereas blastocyst formation rate in the 5 µg/ml treatment group was significantly reduced. UBC9 regulates SUMO-1 content in mature pig oocytes in vitro, which affects oocyte maturation rate, viability, apoptotic genes expression and embryo development after fertilization.

Thyroid antibody status exerts insignificant effect on lymph node metastasis of thyroid cancer.

BACKGROUND: To investigate whether high thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb) levels are associated with increased risk of lymph node metastasis (LNM) of thyroid cancer. METHODS: Data of 2,352 patients who committed thyroidectomy from January 2018 to December 2018 at our institution were retrospectively reviewed. Of which, 806 patients diagnosed with thyroid cancer with available data of both TPOAb and TgAb were finally included, and were divided into four groups: (I) TPOAb-/TgAb- (control, n=493), (II) TPOAb+/TgAb- (n=96), (III) TPOAb-/TgAb+ (n=104), and (IV) TPOAb+/TgAb+ (n=113). The demographic and clinicopathological data were analyzed. RESULTS: Compared to control, significantly less extrathyroidal invasions were identified in TPOAb+ and/or TgAb+ patients (P<0.05), while no significant differences for tumor size, multifocality, or central/lateral neck LNM rate were found for TPOAb+ and/or TgAb+ groups (all P>0.05). Compared to control, significantly more lymph nodes were removed during neck dissection (P<0.05), but there were no significant differences for the number or size of lymph nodes involved (all P>0.05) for TPOAb+ and/or TgAb+ patients. TPOAb+ and/or TgAb+ were not identified as risk factors or protect factors of LNM of thyroid cancer in Logistic regression analyses. CONCLUSIONS: In the present study, we demonstrated that anti-thyroid peroxidase and thyroglobulin antibodies are not associated with increased risk of lymph node metastasis of thyroid cancer.

Genome sequence characterization of canine parvoviruses prevalent in the Sichuan province of China.

Canine parvovirus 2 (CPV-2) infection is responsible for large numbers of animal deaths worldwide and is one of the most dangerous infectious diseases in young puppies. Twenty-four rectal swabs were collected from dogs with clinical signs of vomiting and haemorrhagic diarrhoea and were initially verified to be infected with CPV-2 using colloidal gold test strips. From the 24 CPV-positive samples, complete genome of 5050-5054 nucleotides was sequenced with a next-generation sequencing platform. Characteristics of the Open Reading Frames from different CPV-2 strains detected in this study were analyzed. Several VP2 point mutations were discovered, and demonstrated the co-circulation of new CPV-2a, new CPV-2b and CPV-2c in Sichuan province of China. The analysis results of the Chinese CPV-2 retrieved from the NCBI nucleotide, showed that new CPV-2a has become the predominant variant in some provinces of China. Phylogenetic analysis of global VP2 and NS1 nucleotide sequences revealed certain correlations among geographical regions, types and circulating time, which lays the foundation for further research concerning the epidemiology, genetic variation, vaccination and molecular evolutionary relationships of the CPV-2 identified at different times and from different regions.

Graphene oxide encapsulated polyvinyl alcohol/sodium alginate hydrogel microspheres for Cu (II) and U (VI) removal.

In this work, a novel sodium alginate (SA)/polyvinyl alcohol (PVA)/graphene oxide (GO) hydrogel microspheres were prepared by a simple method. Sodium alginate was physically crosslinked by Ca2+; GO was encapsulated into the composite to strengthen the hydrogels; PVA played a significant role in well dispersing of GO in SA. The SA/PVA/GO (SPG) hydrogels were employed as an efficient adsorbent for removal of Cu (II) and U (VI) from aqueous solution. Batch experiments with the subject of the pH, initial metal ion concentration, competing ions and contact time were investigated. Structure characterization was successfully conducted by FTIR, SEM, EDX, BET and XPS. Furthermore, the sorption kinetics of Cu2+ and UO22+ followed pseudo-second order model and exhibited 3-stage intraparticle diffusion model. Equilibrium data were best described by Langmuir model and the obtained maximum adsorption capacities of SPG hydrogel microspheres for Cu2+ and UO22+ were 247.16 and 403.78 mg/g, respectively. The difference in adsorption capacity can be confirmed by the percentage of elements in EDX spectra and the intension of peak of elements in XPS spectra. The SPG sorbent exhibited excellent reusability after 5 adsorption-desorption cycles. All results suggested that the prepared adsorbents could be considered as effective and promising materials for removal of Cu (II) and U (VI) in wastewater.

Preparation of a novel polyacrylic acid and chitosan interpenetrating network hydrogel for removal of U(vi) from aqueous solutions.

A clean and simple method has been developed for preparation of interpenetrating polymer networks using polyacrylic acid (PAA) and chitosan (CS) for extraction of uranium from polluted water. The peak of Fourier transform infrared spectroscopy (FTIR) occurred at 928 cm-1 indicating combination of uranium and PAA/CS. The energy dispersive X-ray (EDX) and the scanning electron microscope (SEM) studies illustrated the formation of a crosslinking structure and excellent binding ability of uranium on PAA/CS. The maximum adsorption capacity was 289.6 mg g-1 calculated using the equation of the Langmuir model. The adsorption capacity reached a plateau at pH 4 and the sorption process fits the pseudo-second-order model well. The PAA/CS composite has stability of reuse, with the adsorbent capacity decreasing slowly with increasing usage frequency. The experimental results confirm that the PAA/CS hydrogel could be a novel alternative for highly efficient removal of uranium from wastewater.

A comparison of transcriptomic patterns measured in the skin of Chinese fine and coarse wool sheep breeds.

We characterised wool traits, and skin gene expression profiles of fine wool Super Merino (SM) and coarse wool Small Tail Han (STH) sheep. SM sheep had a significantly higher total density of wool follicles, heavier fleeces, finer fibre diameter, and increased crimp frequency, staple length and wool grease (lanolin) production. We found 435 genes were expressed at significantly different levels in the skin of the two breeds (127 genes more highly in SM and 308 genes more highly in STH sheep). Classification of the genes more highly expressed in SM sheep revealed numerous lipid metabolic genes as well as genes encoding keratins, keratin-associated proteins, and wool follicle stem cell markers. In contrast, mammalian epidermal development complex genes and other genes associated with skin cornification and muscle function were more highly expressed in STH sheep. Genes identified in this study may be further evaluated for inclusion in breeding programs, or as targets for therapeutic or genetic interventions, aimed at altering wool quality or yield. Expression of the lipid metabolic genes in the skin of sheep may be used as a novel trait with the potential to alter the content or properties of lanolin or the fleece.