LncRNA TUG1 influences osteoblast proliferation and differentiation through the Wnt/ß-catenin signaling pathway.

OBJECTIVE: Long non-coding ribonucleic acids (lncRNAs) play a vital role in bone development, but the function of lncRNA taurine up-regulated gene 1 (TUG1) in osteoblast proliferation and differentiation is still unknown. MATERIALS AND METHODS: The expression of TUG1 and the messenger RNA (mRNA) expressions of the Wnt/ß-catenin signaling pathway markers [Runt-related transcription factor 2 (Runx2), Frizzled-2, axis inhibition protein 2 (Axin 2) and ß-catenin] at 0 d, 1 d, 7 d and 14 d after in vitro culture of osteoblasts were detected, respectively, by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The effects of TUG1 on the Wnt/ß-catenin signaling pathway markers and osteoblast proliferation and differentiation were studied through the silencing of TUG1 by short hairpin TUG1 (shTUG1). Furthermore, the effects of the Wnt/ß-catenin signal on osteoblast proliferation and differentiation was verified by Wnt/ß-catenin signal inhibitors. RESULTS: With the continuous differentiation of osteoblasts, the level of TUG1 was significantly increased. The mRNA levels of the Wnt/ß-catenin signaling pathway markers (Runx2, Frizzled-2, Axin 2 and ß-catenin) also showed the same increasing trend. ShTUG1 notably reduced the activity of alkaline phosphatases (ALPs), the levels of osteocalcin and osteopontin and osteoblast proliferation activity. In addition, the silencing of TUG1 by shTUG1 resulted in significant reductions in the proteins of the Wnt/ß-catenin signaling pathway markers (Runx2, Frizzled-2, Axin 2 and ß-catenin), and Wnt/ß-catenin inhibitors markedly reduced osteoblast proliferation and differentiation activity. CONCLUSIONS: LncRNA TUG1 inhibition can suppress the Wnt/ß-catenin signaling pathway and reduce osteoblast proliferation and differentiation.

Effect of in ovo feeding of vitamin C on antioxidation and immune function of broiler chickens.

Hypoimmunity and numerous stresses are two major challenges in broiler industry. Nutrient intervention at the specific time of embryonic stage is a feasible way to improve animal performance. This study was conducted to investigate the possible effects of in ovo feeding (IOF) of vitamin C at embryonic age 15th day (E15) on growth performance, antioxidation and immune function of broilers. A total of 240 broiler fertile eggs were randomly divided into two groups (0 and 3 mg injected dose of vitamin C at E15), and new-hatched chicks from each treatment were randomly allocated into six replicates with 10 chicks per replicate after incubation. The results indicated that in ovo vitamin C injection improved the hatchability (P < 0.05) and increased immunoglobulin M (IgM) (at the broiler's age 1st day, D1), IgG and IgM concentrations (D21), as well as lysozyme activity (D21, P < 0.05) and total antioxidant capacity (D42, P < 0.01) in plasma of broilers. On D21, the splenic expression level of DNA methyltransferase 1 (DNMT1) was up-regulated in vitamin C (VC) group, whereas interleukin (IL)-6, interferon-γ, ten-eleven translocation protein 1 and thymine-DNA glycosylase were down-regulated (P < 0.05). On D42, in ovo vitamin C injection up-regulated splenic expression levels of DNMT1, DNA methyltransferase 3B (DNMT3B) and growth arrest and DNA-damage-inducible protein beta (P < 0.05), whereas down-regulated splenic expression levels of IL-6, tumour necrosis factor-α and methyl-CpG-binding domain protein 4 (P < 0.05). Our findings suggested that IOF of 3 mg vitamin C at E15 could improve, to some extent, the antioxidant activity and immune function in plasma, corresponding with the lower expression of pro-inflammatory cytokines in spleen. However, IOF of vitamin C leading to the changes in the expression of DNA methyltransferases and demethylases may suggest an increased trend of DNA methylation level in spleen and whether DNA methylation variation is associated with the lower expression of pro-inflammatory cytokines in spleen warrants future study.

Novel anti-HIV lancilactone C and related triterpenes from Kadsura lancilimba.

Three new triterpene lactones, lancilactones A (1), B (2), and C (3), together with the known kadsulactone A (4), were isolated from the stems and roots of Kadsura lancilimba. Their structures and stereochemistries were determined primarily from mass and NMR spectral data. Compound 3 inhibited HIV replication with an EC50 value of 1.4 microg/mL and a therapeutic index of greater than 71.4.