The potential biological functions of circular RNAs during the initiation of atresia in pig follicles.

The specific expression profile and function of circular RNAs (circRNAs) in mammalian ovarian follicles, especially during the atresia process, are unclear. In this study, genome-wide deep circRNA sequencing was applied to screen circRNAs in healthy and early atretic antral follicles in pig ovaries. A total of 40,567 distinct circRNAs were identified in follicles, among which 197 circRNAs (108 upregulated and 89 downregulated) were significantly shifted during the early atresia process. Most differentially expressed circRNAs (DECs) lacked protein-coding potential. Annotation analysis of the DECs revealed 162 known host genes, or noncoding RNAs, and 10 intergenic regions. The key pathways in which these host genes are involved include the focal adhesion-PI3K-Akt-mTOR signaling pathway, vascular endothelial growth factor A (VEGFA)-vascular endothelial growth factor receptor 2 signaling pathway and transforming growth factor-beta signaling pathway. Further comparison analysis between host genes of DECs and the differentially expressed linear messenger RNA transcripts revealed the cotranscription of circRNAs and their linear mRNAs in inhibin beta units (INHBA and INHBB), glutathione S-transferase (GSTA1), and VEGFA. In addition, we predicted 196 pairs of potential circRNA-micro RNA (miRNA) interactions among 77 DECs and 101 porcine miRNAs. We have identified 16 functional miRNAs by comparing the 101 miRNAs to the functional miRNAs reported in mammal ovarian follicle atresia and granulosa cell apoptosis studies. Our study adds new knowledge to circRNA distribution profiles in pig ovarian follicles, offers a valuable reference for transcriptomic profiles in the initiation of follicular atresia, highlights warranted circRNAs for further functional investigation, and provides possible biomarkers for ovarian dysfunctions.

Downregulation of miR-1294 associates with prognosis and tumor progression in epithelial ovarian cancer.

OBJECTIVE: Increasing evidence has revealed that microRNAs (miRNAs) act as key players in the regulation of tumor growth and metastasis in epithelial ovarian cancer (EOC). However, the clinical role and functional effects of miR-1294 in EOC remain unknown. PATIENTS AND METHODS: We examined the expression of miR-1294 in 69 cases of EOC tissues and cell lines by quantitative Real-time polymerase chain reaction (qRT-PCR). The associations of miR-1294 expression with clinicopathologic features and overall survival of EOC patients were analyzed. Biological functional effects of miR-1294 expression on cell growth were analyzed using Cell Counting Kit-8 (CCK8) assays and flow cytometry assays in vitro. RESULTS: In the present study, we identified that miR-1294 expression was lower in 76 specimens of EOC compared to adjacent normal tissues. Lower miR-1294 expression was related to FIGO stage, lymph node metastasis and shorter overall survival rate in EOC patients. Multivariate Cox analysis demonstrated that miR-1294 expression was an independent prognostic indicator of EOC patients. Gain function assays showed that miR-1294 overexpression inhibited cell proliferation and cell cycle progression in EOC. CONCLUSIONS: Our results indicated that miR-1294 acted as a prognostic biomarker and potential target of EOC treatment.

Effects of first feed intake time on growth performance, nutrient apparent metabolic rate and intestinal digestive enzyme activities in broilers.

OBJECTIVE: This experiment studied the effects of first feed intake time post-hatch on growth performance, nutrient apparent metabolic rate and intestinal digestive enzyme activities in broilers. METHODS: Two thousand five hundred and twenty LingNan Yellow broilers were randomly allotted to seven treatments with six replicates of 60 each. The only experimental factor was the first feed intake time which was 18, 24, 30, 36, 42, 48, and 54 hours after hatching. The whole experiment lasted for 21 days. RESULTS: During the whole period, the 30 h treatment had the best body weight and average daily gain (p<0.05), followed by the 24 h group performance optimization. Also, the 30 h group was observed to have the best apparent metabolic rate for ether extract (p<0.05) and crude protein (p<0.05) and the highest activities of amylase, lipase and trypsin in small intestine. And the 24 h group was second only to the 30 h group in terms of the above two measures. CONCLUSION: These results indicated that the appropriate first feeding time of LingNan Yellow broilers was 24 to 30 hours after hatching.

Adsorptive separation of hemoglobin by molecularly imprinted chitosan beads.

A simply hemoglobin (Hb) molecularly imprinted polymer (MIP) was prepared using Hb as the imprinted molecule, acrylamide as the functional monomer and cross-linked chitosan beads as the supporting matrix. The MIP was achieved by entrapment of the selective soft polyacrylamide gel in the pores of the cross-linked chitosan beads by letting acrylamide monomer and the protein diffuse into the pores of chitosan beads before starting the polymerization. The chitosan beads were freed from the surrounding polyacrylamide gel by washing. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms. Langmuir analysis showed that an equal class of adsorption was formed in the MIP and the adsorption equilibrium constant and the maximum adsorption capacity were evaluated. The MIP has much higher adsorption capacity for Hb than the non-imprinted polymer with the same chemical composition, and the MIP also has a higher selectivity for the imprinted molecule. The MIP can be reused in an easy way and the reproduction coefficient was approximately 100% at low concentration.

Pulsed electrospray for mass spectrometry.

Pulsed electrospray has been developed and been reported for the first time. This new technique is based on the principle of pulsed ion sources, combined with the conventional electrospray. The pulsed ion spray was realized by a homemade pulsed HV circuit and was monitored by a digital microscope and an oscilloscope. Results show that the pulsed ESI device can be operated under proper conditions for a clear on-and-off spray process and that the device was kept in good electric contact for electrospray when pulsed HV was on. A pulsed ion current and pulsed mass spectra can be achieved with this pulsed ESI device. Furthermore, it has been noted that, under the same conditions (i.e., shape and size of sprayer tip, distance from sprayer tip to sampling nozzle, and other parameters for mass spectrometer), stable electrospray could be obtained for lower flow rates with a pulsed spray device. This experimental fact indicates the possible reduction in the total sample consumed could be realized by exploiting this novel design.