miR-206a-3p suppresses the proliferation and differentiation of chicken chondrocytes in tibial dyschondroplasia by targeting BMP6.

The poultry skeletal system serves multiple functions, not only providing structural integrity but also maintaining the balance of essential minerals such as calcium and phosphorus. However, in recent years, the consideration of skeletal traits has been overlooked in the selective breeding of broilers, resulting in an inadequate adaptation of the skeletal system to cope with the rapid increase in body weight. Consequently, this leads to lameness and bone diseases such as tibial dyschondroplasia (TD), which significantly impact the production performance of broilers. Accumulating evidence has shown that microRNAs (miRNA) play a crucial role in the differentiation, formation, and disease of cartilage. However, the miRNA-mediated molecular mechanism underlying chicken TD formation is still poorly understood. The objective of this study was to investigate the biological function and regulatory mechanism of miRNA in chicken TD formation. Based on transcriptome sequencing of tibial cartilage in the healthy group and TD group, miR-206a-3p was found to be highly expressed in TD cartilage. The function of miR-206a-3p was explored through the transfection test of miR-206a-3p mimics and miR-206a-3p inhibitor. In this study, we utilized qRT-PCR, CCK-8, EdU, western blot, and flow cytometry to detect the proliferation, differentiation, and apoptosis of chondrocytes. The results revealed that miR-206a-3p suppressed the proliferation and differentiation of TD chondrocytes while promoting their programmed cell death. Furthermore, through biosynthesis and dual luciferase assays, it was determined that BMP6 was the direct target gene of miR-206a-3p. This finding was further supported by rescue experiments which confirmed the involvement of BMP6 in the regulatory pathway governed by miR-206a-3p. Our results suggest that miR-206a-3p can inhibits the proliferation and differentiation promote apoptosis through the target gene BMP-6 and suppressing the Smad2/3 signaling pathway in chicken TD chondrocytes.

Thiram exposure induces tibial dyschondroplasia in broilers via the regulation effect of circ_003084/miR-130c-5p/BMPR1A crosstalk on chondrocyte proliferation and differentiation.

Thiram, an agricultural insecticide, has been demonstrated to induce tibial dyschondroplasia (TD) in avian species. Circular RNA (circRNAs), a novel class of functional biological macromolecules characterized by their distinct circular structure, play crucial roles in various biological processes and diseases. Nevertheless, the precise regulatory mechanism underlying non-coding RNA involvement in thiram-induced broiler tibial chondrodysplasia remains elusive. In this study, we established a broiler model of thiram exposure for 10 days to assess TD and obtain a ceRNA network by RNA sequencing. By analyzing the differentially expressed circRNAs network, we id entify that circ_003084 was significantly upregulated in TD cartilage. Elevated circ_003084 inhibited TD chondrocytes proliferation and differentiation in vitro but promote apoptosis. Mechanistically, circ_003084 competitively binds to miR-130c-5p and prevents miR-130c-5p to decrease the level of BMPR1A, which upregulates the expression of apoptosis genes Caspase 3, Caspase 9, Bax and Bcl2, and finally facilitates cell apoptosis. Taken together, these findings imply that circ_003084/miR-130c-5p/BMPR1A interaction regulated TD chicken chondrocyte proliferation, apoptosis, and differentiation. This is the first work to reveal the mechanism of regulation of circRNA-related ceRNA on thiram-induced TD, offering a key reference for environmental toxicology.

Probiotic mediated intestinal microbiota and improved performance, egg quality and ovarian immune function of laying hens at different laying stage.

In order to investigate the effects of dietary probiotics supplementation on laying performance, egg quality, serum hormone levels, immunity, antioxidant, and gut microbiota of layers at different laying stages, a total of 168 Tianfu green shell laying hens (28-day-old) were randomly divided into 2 treatments: a non-supplemented control diet (NC), and diet supplemented with 10 g/kg of probiotics, respectively. Each treatment had 6 replicates with 14 hens per replicate. The feeding trial lasted for 54 weeks. The results showed that the supplementation of probiotics significantly increased the average egg weight, improved egg quality (p < 0.05) and ovarian development. Meanwhile, probiotics increased the serum hormone levels of E2 and FSH, and antioxidant indices T-AOC and T-SOD (p < 0.05) of laying hens at different laying stages (p < 0.05), decreased the expression of proinflammatory factors including IL-1, IL-6 and TNF-α (p < 0.05). Furthermore, using 16S rRNA sequencing, we observed that the addition of probiotics increased the distribution of Firmicutes, Bacteroidota and Synergistota at early laying period. Meanwhile, Bacteroidota, Actinobacteriota, Verrucomicrobiota and Deferribacterota showed an increasing trend at the peak of egg production. The relative abundance of Firmicutes, Desulfobacterota and Actinobacteriota were significantly increased at the late laying period. Moreover, PICRUSt2 and BugBase analysis revealed that at the late laying period, the probiotics supplementation not only enriched many significant gene clusters of the metabolism of terpenoids and polyketide, genetic information processing, enzyme families, translation, transcription, replication and repair, and nucleotide metabolism, but also decreased the proportion of potential pathogenic bacteria. To sum up, these data show that the addition of probiotics not only improves the performance, egg quality, ovarian development and immune function of laying hens at different laying period, but also improves the gut microbiota of layers, thus enhances production efficiency.

Comprehensive analysis of differently expression mRNA and non-coding RNAs, and their regulatory mechanisms on relationship in thiram-induced tibial dyschondroplasia in chicken.

Thiram pollution is one of the main causes of tibial dyschondroplasia (TD) induced by feed sources. Several studies have speculated that miRNA, circRNA and lncRNA may have significant impact on the development of TD, however, the specific mRNAs and noncoding RNAs and their respective regulatory mechanisms and functions in the development of TD have not been explored. Therefore, in this present study, we screened the differentially expressed mRNA, miRNA, circRNA and lncRNA by whole-transcriptome sequencing (RNA-seq) and differentially expressed genes (DEGs) enrichment, as well as constructed the interaction network among the mRNA-miRNA, mRNA-lncRNA and mRNA-miRNA-circRNA. The sequencing results were verified by fluorescence real-time quantitative PCR (RT-qPCR). The results obtained in this study, revealed that the cells were atrophied and disordered in the TD group, and the expression of BMP6, TGF-ß and VEGF were significantly reduced. A total of 141 mRNAs, 10 miRNAs, 23 lncRNAs and 35 circRNAs of DEGs were obtained (p<0.05) Theses DEGs were enriched in the adhere junction and insulin signaling pathways. In addition, the mRNA-miRNA-circRNA network suggested that several pivotal ceRNA showed a regulatory relationship between the transcripts with miRNA, circRNA or lncRNA. Taken together, the results in the present study, represent an insight for further functional research on the ceRNA regulatory mechanism of TD in broilers.

Effect of BMP6 on the proliferation and apoptosis of chicken chondrocytes induced by thiram.

The development of skeleton system is a complex biological process and be regulated by many transcription factors. Previous studies have shown that BMP6 is involved in skeleton development and other cells transforming to chondrocytes, but it is still not known whether do something to tibial dyschondroplasia (TD) broilers chondrocytes. In this study, RT-PCR revealed that the expression level of BMP6 in TD broiler chondrocytes at 7 days age was significantly decreased compared with normal group (P < 0.05). CCK-8 and EdU assay showed that the proliferation of cells transfected with interference BMP6 was significantly decreased compared with control siRNA, while cell proliferation was significantly increased after overexpression of BMP6. Meanwhile, the proportion of G0/G1 phase cells was significantly increased and the proportion of G2/M phase cells was significantly decreased after interference of BMP6 for 48 h in TD chicken chondrocytes (P < 0.05). In addition, flow cytometry analysis exhibited that interference BMP6 significantly increased apoptosis rate and necrotizing rate of cells. In conclusion, these results suggest that BMP6 plays a positive role in the growth and development of TD broiler chondrocytes. Our findings reveal a new target for TD prevention in broiler chickens.

Non-vitamin K antagonist oral anticoagulants versus warfarin in AF patients ≥ 85 years.

BACKGROUND: Atrial fibrillation (AF) prevalence and its risk of stroke rise with ageing. We aimed to investigate the outcomes of NOAC and warfarin in AF patients aged ≥ 85 years. METHODS: This is a retrospective study using Taiwan National Health Insurance Research Database. A total of 15,361 patients aged ≥ 85 years with AF on oral anticoagulants were identified. The end points included ischaemic stroke, intracranial haemorrhage (ICH), major bleeding, all-cause mortality and composite adverse events (ICH or major bleeding or all-cause mortality). Clinical outcomes were compared between each NOAC and warfarin after propensity matching. RESULTS: Before propensity matching, patients taking warfarin were older, more female with more comorbidities than NOACs users. After propensity matching, baseline characteristics did not differ significantly between matched subjects receiving warfarin and each NOAC. Compared to warfarin, dabigatran was associated with a lower risk of ICH (hazard ratio [HR] 0.496), mortality (HR 0.558) and adverse events (HR 0.628), while rivaroxaban was associated with a lower risk of ischaemic stroke (HR 0.781), ICH (HR 0.453), mortality (HR 0.558) and adverse events (HR 0.636). Apixaban was associated with a lower risk of mortality (HR 0.488) and adverse events (HR 0.557) compared to warfarin. (all P < .05). CONCLUSION: For the efficacy, NOACs were associated with a comparable or lower risk of ischaemic stroke compared to warfarin. For adverse events, NOACs were associated with a lower risk of all-cause mortality and composite adverse events. In the elderly AF population, NOACs could be a more favourable choice for stroke prevention.

Screening of Deletion Variants within the Goat PRDM6 Gene and Its Effects on Growth Traits.

By genome-wide association studies, the PRDM6 gene has been shown to affect multiple, apparently unrelated inherited traits, including bone density and body mass index. Therefore, it is considered a potentially pleiotropic gene. In this study, we identified a 12 bp deletion variant (NC_030814.1:rs651603667, g: 79985625-79985636delTTGACTGATCCA) within the PRDM6 gene in a large sample (SBWC goats; n = 1044). All goat samples were collected in Shaanxi province in July 2018. The frequency of the wt allele was higher than the frequency of the del allele, and this mutation polymorphism confirmed to be consistent with the Hardy-Weinberg equilibrium (p > 0.05). Further results showed that in a group of goats in the yearling period (18 months old, n = 567), this deletion variant of the PRDM6 gene was associated with heart girth (p = 0.027), cannon circumference (p = 0.008), chest depth (p = 2.10 × 10-5), chest width (p = 0.004), body height (p = 0.032), body length (p = 0.044) and hip-width (p = 0.014). For adult SBWC goats (36 months old, n = 477), the effects of the 12 bp variation on growth-related traits were found to make no difference. These findings show that the 12 bp deletion within the goat PRDM6 gene plays an important role in the early growth and development of goats. Using the 12 bp mutation, breeders can quickly and effectively select excellent individual goats at an early stage.

Level set contouring for breast tumor in sonography.

The echogenicity, echotexture, shape, and contour of a lesion are revealed to be effective sonographic features for physicians to identify a tumor as either benign or malignant. Automatic contouring for breast tumors in sonography may assist physicians without relevant experience, in making correct diagnoses. This study develops an efficient method for automatically detecting contours of breast tumors in sonography. First, a sophisticated preprocessing filter reduces the noise, but preserves the shape and contrast of the breast tumor. An adaptive initial contouring method is then performed to obtain an approximate circular contour of the tumor. Finally, the deformation-based level set segmentation automatically extracts the precise contours of breast tumors from ultrasound (US) images. The proposed contouring method evaluates US images from 118 patients with breast tumors. The contouring results, obtained with computer simulation, reveal that the proposed method always identifies similar contours to those obtained with manual sketching. The proposed method provides robust and fast automatic contouring for breast US images. The potential role of this approach might save much of the time required to sketch a precise contour with very high stability.