The objectives of this study were to determine how dietary supplementation of N-carbamylglutamate (NCG) and rumen-protected L-arginine (RP-Arg) in nutrient-restricted pregnant Hu sheep would affect (1) maternal endocrine status; (2) maternal, fetal, and placental antioxidation capability; and (3) placental development. From day 35 to day 110 of gestation, 32 Hu ewes carrying twin fetuses were allocated randomly into four groups: 100% of NRC-recommended nutrient requirements, 50% of NRC recommendations, 50% of NRC recommendations supplemented with 20 g/day RP-Arg, and 50% of NRC recommendations supplemented with 5 g/day NCG product. The results showed that in maternal and fetal plasma and placentomes, the activities of total antioxidant capacity and superoxide dismutase were increased (P < 0.05); however, the activity of glutathione peroxidase and the concentration of maleic dialdehyde were decreased (P < 0.05) in both NCG-and RP-Arg-treated underfed ewes. The mRNA expression of vascular endothelial growth factor and Fms-like tyrosine kinase 1 was increased (P < 0.05) in 50% NRC ewes than in 100% NRC ewes, and had no effect (P > 0.05) in both NCG-and RP-Arg-treated underfed ewes. A supplement of RP-Arg and NCG reduced (P < 0.05) the concentrations of progesterone, cortisol, and estradiol-17β; had no effect on T 4 /T 3 ; and improved (P < 0.05) the concentrations of leptin, insulin-like growth factor 1, tri-iodothyronine (T 3 ), and thyroxine (T 4 ) in serum from underfed ewes. These results indicate that dietary supplementation of NCG and RP-Arg in underfed ewes could influence maternal endocrine status, improve the maternal-fetal-placental antioxidation capability, and promote fetal and placental development during early-to-late gestation.

Aim. To explore the pharmacological mechanism of Xiaoyao powder (XYP) on anovulatory infertility by a network pharmacology approach. Method. Collect XYP's active compounds by traditional Chinese medicine (TCM) databases, and input them into PharmMapper to get their targets. Then note these targets by Kyoto Encyclopedia of Genes and Genomes (KEGG) and filter out targets that can be noted by human signal pathway. Get the information of modern pharmacology of active compounds and recipe's traditional effects through databases. Acquire infertility targets by Therapeutic Target Database (TTD). Collect the interactions of all the targets and other human proteins via String and INACT. Put all the targets into the Database for Annotation, Visualization, and Integrated Discovery (DAVID) to do GO enrichment analysis. Finally, draw the network by Cytoscape by the information above. Result. Six network pictures and two GO enrichment analysis pictures are visualized. Conclusion. According to this network pharmacology approach some signal pathways of XYP acting on infertility are found for the first time. Some biological processes can also be identified as XYP's effects on anovulatory infertility. We believe that evaluating the efficacy of TCM recipes and uncovering the pharmacological mechanism on a systematic level will be a significant method for future studies.

As an important type of noncoding RNA molecules, long non-coding RNAs (lncRNAs) act as versatile players in various biological processes. However, little is known about lncRNA regulators during sheep muscle growth. To explore functional lncRNAs during sheep muscle growth, we systematically investigated lncRNAs using strand-specific Ribo-Zero RNA sequencing at three key developmental stages in Hu sheep. A total of 6924 lncRNAs were obtained, and the differentially expressed lncRNAs and genes were screened from (control vs. experiment) fetus vs. lamb, lamb vs. adult, and fetus vs. adult comparisons, respectively. The quantitative real-time polymerase chain reaction (qRT-PCR) analysis results correlated well with the sequencing data. Moreover, functional annotation analysis based on the Gene Ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) databases showed that the target genes of the differentially expressed lncRNAs were significantly enriched in organ morphogenesis, skeletal system development as well as response to stimulus and some other terms related to muscle. Furthermore, a co-expression network of the differentially expressed target genes and lncRNAs was constructed and well-known muscle growth regulators such as retrotransposon-like 1 and Junctophilin-2 were included. Finally, we investigated the expression profiles of seven lncRNAs and their target genes, and found that they played vital roles in muscle growth. This study extends the sheep muscle lncRNA database and provides novel candidate regulators for future genetic and molecular studies on sheep muscle growth, which is helpful for optimizing the production of mutton.

To investigate the pharmacological mechanism of Guizhi Fuling Wan (GFW) in the treatment of uterine fibroids, a network pharmacology approach was used. Information on GFW compounds was collected from traditional Chinese medicine (TCM) databases, and input into PharmMapper to identify the compound targets. Genes associated with uterine fibroids genes were then obtained from the GeneCards and Online Mendelian Inheritance in Man databases. The interaction data of the targets and other human proteins was also collected from the STRING and IntAct databases. The target data were input into the Database for Annotation, Visualization and Integrated Discovery for gene ontology (GO) and pathway enrichment analyses. Networks of the above information were constructed and analyzed using Cytoscape. The following networks were compiled: A compound-compound target network of GFW; a herb-compound target-uterine fibroids target network of GWF; and a compound target-uterine fibroids target-other human proteins protein-protein interaction network, which were subjected to GO and pathway enrichment analyses. According to this approach, a number of novel signaling pathways and biological processes underlying the effects of GFW on uterine fibroids were identified, including the negative regulation of smooth muscle cell proliferation, apoptosis, and the Ras, wingless-type, epidermal growth factor and insulin-like growth factor-1 signaling pathways. This network pharmacology approach may aid the systematical study of herbal formulae and make TCM drug discovery more predictable.

Bone marrow mesenchymal stem cells (BMSCs), which are well characterized and widely utilized adult stem cells, encompass the capacity to commit to a variety of cell types. This study was conducted to develop an effective way to induce goat BMSCs (gBMSCs) to transdifferentiate toward putative male germ cells by overexpressing STRA8 (stimulated by RA-8), BOULE (also called BOLL), and DAZL (deleted in azoospermia-like). First, we found that the expression levels of these 3 genes gradually increased during development of the goat testis from 10 days postnatal to 8 months old. Therefore, we hypothesized that overexpressing these genes might contribute to the transdifferentiation of gBMSCs toward germ cells. We then overexpressed, separately and in combination, STRA8, BOULE, and DAZL in gBMSCs. Our results showed that a small population of transfected gBMSCs transdifferentiated into early goat germ cell-like cells and that these cells expressed primordial germ cell specification genes STELLA (also known as DPPA3, developmental pluripotency associated 3) and C-KIT (tyrosine kinase receptor) as well as premeiotic genes MVH (mouse vasa homolog), DAZL, BOULE, STRA8, PIWIL2 (piwi-like RNA-mediated gene silencing 2), and RNF17 (ring finger protein 17). Importantly, results from quantitative reverse transcription polymerase chain reaction, immunofluorescence, and Western blot analysis showed that the meiotic marker synaptonemal complex protein 3 (SCP3) significantly increased in transfected cells compared to untransfected control cells ( P < .05). Additionally, the co-overexpression group cells had the highest SCP3 messenger RNA and protein expression levels, which indicated that 3-gene co-overexpression had the highest potential to transdifferentiate gBMSCs to germ cells. Taken together, these results demonstrate that the overexpression of STRA8, BOULE, and DAZL was able to promote the transdifferentiation of gBMSCs to early goat germ cell-like cells in vitro, which probably enhanced maturation and progression through meiosis. This approach would be important to generating gametes for future basic science as well as for potential clinical applications.

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Cancer stem cells (CSCs) are a subpopulation of cancer cells believed to be implicated in cancer initiation, progression, and recurrence. Here, we report that ectopic expression of zinc finger E-box binding homeobox 1 protein (ZEB1) results in the acquisition of CSC properties by breast cancer cells, leading to tumor initiation and progression in vitro and in vivo. The neurogenin 3 gene (Ngn3) is a bona fide target of ZEB1, and its repression is a key factor contributing to ZEB1-induced cancer cell stemness. ZEB1 suppressed Ngn3 transcription by forming a ZEB1/DNA methyltransferase (DNMT)3B/histone deacetylase 1 (HDAC1) complex on the Ngn3 promoter, leading to promoter hypermethylation and gene silencing. The rescue of Ngn3 expression attenuated ZEB1-induced cancer stemness and symmetric CSC division. Immunohistological analysis of human breast cancer specimens revealed a strong inverse relationship between ZEB1 and NGN3 protein expression. Thus, our findings suggest ZEB1-mediated silencing of Ngn3 is required for breast tumor initiation and maintenance. Targeted therapies against the ZEB1/Ngn3 axis may be highly valuable for the prevention and treatment of breast cancer.