Identification and analysis of differentially expressed microRNAs in endometrium to explore the regulation of sheep fecundity.

BACKGROUND: MicroRNAs (miRNAs) play an important regulatory role in mammalian reproduction. Currently, most studies are primarily concentrated on ovarian miRNAs, ignoring the influence of endometrial miRNAs on the fecundity of female sheep. To uncover potential regulators of sheep fecundity, RNA-seq was used to comparatively analyze miRNA expression profiles of endometrium between high prolificacy sheep (HP, litter size = 3) and low prolificacy sheep (LP, litter size = 1) with FecB genotype. RESULTS: Firstly, genomic features of miRNAs from endometrium were analyzed. Furthermore, 58 differentially expressed (DE) miRNAs were found in the endometrium of Hu sheep with different litter size. A co-expression network of DE miRNAs and target genes has been constructed, and hub genes related litter size are included, such as DE miRNA unconservative_NC_019472.2_1229533 and unconservative_NC_019481.2_1637827 target to estrogen receptor α (ESR1) and unconservative_NC_019481.2_1637827 targets to transcription factor 7 (TCF7). Moreover, functional annotation analysis showed that the target genes (NRCAM and NEGR1) of the DE miRNAs were significantly enriched in cell adhesion molecules (CAMs) signaling pathway, which was related to uterine receptivity. CONCLUSION: Taken together, this study provides a new valuable resource for understanding the molecular mechanisms underlying Hu sheep prolificacy.

Repeated Superovulation Accelerates Primordial Follicle Activation and Atresia.

For humans, ARTs (assisted reproductive technologies) have become the most effective method to treat subfertility/infertility in clinic. To obtain enough oocytes during ART, ovarian stimulation is performed by exogenous hormones, and some patients undergo several ovarian stimulation cycles. Although some adverse effects of ARTs on women and offspring are reported, few studies are focused on the effects of multiple superovulation on ovarian reserve. In the present study, we found that repeated superovulation significantly reduced primordial follicle number and the serum AMH. Compared to the decreased antral follicle number, the expression of genes related to primordial follicle activation, such as Foxo3, Akt, and Rptor, and the atretic follicle number in ovaries were increased by superovulation times. We further found that repeated superovulation reduced the plasma level of FSH, LH, and estradiol, and increased the expression of genes related to apoptosis (Bax, Casp3 (caspase-3), Casp8, and Casp9) in granulosa cells, providing evidence that repeated superovulation disrupted the balance between survival and death in granulosa cells. In summary, our results suggest that repeated superovulation has adverse effects on folliculogenesis.

Expression and functional analysis of the Follistatin-like 3 (FSTL3) gene in the sheep ovary during the oestrous cycle.

Follistatin-like 3 (FSTL3) is a regulator of cellular apoptosis and was previously identified via RNA-Seq to be associated with follicular development in mammalian ovaries. However, the mechanism underlying the FSTL3 regulation of oestrus in sheep remained poorly understood. In this study, the oestrogen (E2) and progesterone (P4) concentrations in blood were detected, and the expression level and functional analysis of FSTL3 in the ovary were studied during the different reproductive stage in Aohan fine wool sheep (seasonal breeding breed in China). The concentrations of E2 and P4 at the anestrus were significantly lower compared to dioestrus, proestrus and oestrus stages. Higher expression levels of FSTL3 were observed in the sheep ovary, hypothalamus, and thyroid. During different reproductive stages, higher expression levels were found during the stages of dioestrus and proestrus, while lower levels were found during the oestrus and anestrus stages. Functional analysis of FSTL3 was performed in primary granulosa cells (GCs) of sheep. The concentration of E2 increased significantly after RNAi interference of FSTL3, while the P4 level decreased. FSTL3 can decrease P4 levels, which might be involved in mediating oestrous cycle in sheep.

Eco-friendly and durable PCPS nanoparticles for the effective separation of oil-water emulsions.

Oil pollution is one of the main environmental problems that is attracting increasing attention from people. In this study, a new composite, namely a PNIPAm-Clay-γPGA-SiO2 (PCPS) nanomaterial, was prepared through chemical modification. The material exhibited excellent separation efficiencies for both oil-in-water (O/W) and water-in-oil (W/O) emulsions. The maximum separation efficiency of the W/O emulsion reached 98.70%, while that of the O/W emulsion was 99.23%, and the average separation fluxes were 107.44 L m-2 h-1 and 1529.34 L m-2 h-1, respectively. The superhydrophobicity of the PCPS nanoparticles could be maintained under strong acid/alkali conditions for over 30 days. The high separation efficiency could be maintained even after 7 cycles, indicating the long-term availability of the material. Furthermore, the PCPS nanoparticles showed excellent biocompatibility due to the presence of γ-polyglutamic acid (γPGA) and poly(N-isopropylacrylamide) (PNIPAm). The properties of strong acid/alkali and thermal stabilities, recyclability, and biocompatibility gave the material great potential for applications.

Strip rotary tillage with a two-year subsoiling interval enhances root growth and yield in wheat.

Excessive tillage and soil compaction threaten the sustainable farmlands in the Huang-Huai-Hai Plains of China. Our study explores tillage practices to improve soil and root ecology and promote productivity in the winter wheat fields. We tested the impact of plowing, rotary, strip rotary tillage and strip rotary tillage with a two-year subsoiling interval (SRS) on wheat yield and root quality. SRS decreased soil bulk density compared with other treatments, resulting in lower soil penetration resistance. Root morphology and weight density decreased with the increased soil depth and was higher in SRS. Moreover, SRS increased the indoleacetic acid and trans zeatin riboside levels corresponding to greater TTC reduction activities, the total and active absorption root area. SRS increased the superoxide dismutase and catalase activities and soluble protein concentration and decreased the malondialdehyde concentration. The first two factors extracted using 11 root attributes in various soil layers through principal component analysis were selected as the integrated indicators for the minimum data set, and their integrated score was calculated to quantify the root quality. Our study suggests that SRS could significantly improve root morphology and enhance the root activity in subsoil layers, thus, delaying root senescence and increasing winter wheat yield.

Identification of differentially expressed genes affecting hair and cashmere growth in the Laiwu black goat by microarray.

Goats are an important source of fibers. In the present study microarray technology was used to investigate the potential genes primarily involved in hair and cashmere growth in the Laiwu black goat. A total of 655 genes differentially expressed in body (hair­growing) and groin (hairless) skin were identified, and their potential association with hair and cashmere growth was analyzed. The majority of genes associated with hair growth regulation could be assigned to intracellular, intracellular organelle, membrane­bound vesicle, cytoplasmic vesicle, pattern binding, heparin binding, polysaccharide binding, glycosaminoglycan binding and cytoplasmic membrane­bound vesicle categories. Numerous genes upregulated in body compared with groin skin contained common motifs for nuclear factor 1A, Yi, E2 factor (E2F) and cyclic adenosine monophosphate response element binding (CREB)/CREBß binding sites in their promoter region. The promoter region of certain genes downregulated in body compared with groin skin contained three common regions with LF­A1, Yi, E2F, Collier/Olfactory­1/early B­cell factor 1, peroxisome proliferator­activated receptor α or U sites. Thus, the present study identified molecules in the cashmere­bearing skin area of the Laiwu black goat, which may contribute to hair and cashmere traits.

Characterization and comparative profiling of ovarian microRNAs during ovine anestrus and the breeding season.

BACKGROUND: Seasonal estrus is a critical limiting factor of animal fecundity, and it involves changes in both ovarian biology and hormone secretion in different seasons. Previous studies indicate that two classes of small RNAs (miRNAs and piRNAs) play important regulatory roles in ovarian biology. To understand the roles of small RNA-mediated post-transcriptional regulation in ovine seasonal estrus, the variation in expression patterns of ovarian small RNAs during anestrus and the breeding season were analyzed using Solexa sequencing technology. In addition, reproductive hormone levels were determined during ovine anestrus and the breeding season. RESULTS: A total of 483 miRNAs (including 97 known, 369 conserved and 17 predicated novel miRNAs), which belong to 183 different miRNA families, were identified in ovaries of Tan sheep and Small Tail Han (STH) sheep. Compared with the three stages of the breeding season, 25 shared significantly differentially expressed (including 19 up- and six down-regulated) miRNAs were identified in ovine anestrus. KEGG Pathway analysis revealed that the target genes for some of the differentially expressed miRNAs were involved in reproductive hormone related pathways (e.g. steroid biosynthesis, androgen and estrogen metabolism and GnRH signaling pathway) as well as follicular/luteal development related pathways. Moreover, the expression of the differentially expressed miRNAs and most of their target genes were negatively correlated in the above pathways. Furthermore, the levels of estrogen, progesterone and LH in ovine anestrus were significantly lower than those in the breeding season. Combining the results of pathway enrichment analysis, expression of target genes and hormone measurement, we suggest that these differentially expressed miRNAs in anestrus might participate in attenuation of ovarian activity by regulating the above pathways. Besides miRNAs, a large and unexpectedly diverse set of piRNAs were also identified. CONCLUSIONS: The miRNA profiles of ovine ovaries in anestrus were presented for the first time. The identification and characterization of miRNAs that are differentially expressed between ovine anestrus and the breeding season will help understanding of the role of miRNAs in the regulation of seasonal estrus, and provides candidates for determining miRNAs which could be potentially used to regulate ovine seasonal estrus.

[Internal fixation treatment of multiple rib fractures with absorbable rib-connecting-pins].

OBJECTIVE: To study the indications, methods, and therapeutic effect of absorbable rib-connecting-pins fixation in the treatment of multiple rib fractures. METHODS: Between March 2007 and September 2009, 40 patients with multiple rib fractures received internal fixation with absorbable rib-connecting-pins, including 8 one-side flail chest and 1 two-side flail chest. There were 32 males and 8 females with an average age of 39.8 years (range, 25-72 years). The injury was caused by traffic accident in 32 cases, falling from height in 6 cases, and blunt hitting in 2 cases. Preoperatively, imaging data of the chest X-ray or spiral CT three-dimensional (3D) examination showed that all patients had multiple ribs fractures and displacement. The number of fractured ribs was 4-10 (median, 6), and the fracture location ranged from the 2nd to the 10th ribs. Of them, 28 cases were accompanied by hemathorax, pneumothorax or hemopneumothorax; 5 cases by thoracic organ injury; and 10 cases by other part trauma. The time from injury to hospitalization was less than 1 day in 26 cases, 1-3 days in 12 cases, and 3-6 days in 2 cases, and the time from hospitalization to operation was 3 hours to 3 days (mean, 1.2 days). RESULTS: The median fixation rib number was 5 (range, 3-8). The mean operative time, the time in bed, and hospitalization days were 32 minutes (range, 15-50 minutes), 4.5 days (range, 2-7 days), and 11.2 days (range, 5-18 days), respectively. All incisions healed by first intention. No pulmonary infection, pulmonary atelectasis, intrathoracic infection or other complications occurred. All cases were followed up 6-12 months (mean, 8 months). PaO2 [(86.6 +/- 2.2) mmHg (1 mm Hg = 0.133 kPa)] and SpO2 (97.2% +/- 0.6%) at 2 hours after operation were obviously improved when compared with preoperative ones (PaO2 (53.6 + 4.7) mm Hg and SpO2 (86.2% + 1.8%)], showing significant differences (t = 2.971, P = 0.005; t = 2.426, P = 0.020). The chest X-ray films or spiral CT 3D indicated that fracture of rib healed within 3-6 months (mean, 4.5 months) after operation. CONCLUSION: Severe collapsed chest wall or flail chest caused by fracture of multiple ribs should be treated by absorbable rib-connecting-pins, which is a simple, firm, and effective method.