Testosterone derived from testicular Leydig cells (LCs) is important for male sheep, and the testis is susceptible to external temperature. The present study aimed to explore the alleviating effect of selenomethionine (Se-Met) on heat-induced injury in Hu sheep LCs. Isolated LCs were exposed to heat (41.5 degrees C, heat exposure, HE) or not (37 degrees C, nonheat exposure, NE), and cells in NE and HE were treated with 0 (C) or 8 mu mol/L (S) Se-Met for 6 h. Cell viability, testosterone level, and the expression of GPX1, HSD3B, apoptosis-related genes and p38 mitogen-activated protein kinase (p38MAPK)/heat shock protein beta-1 (HSPB1) pathway were examined. The results showed that Se-Met increased GPX1 expression (NE-S vs. NE-C: 2.28-fold; HE-S vs. HE-C: 2.36-fold, p < 0.05) and alleviated heat-induced decrease in cell viability (HE-S vs. HE-C: 1.41-fold; HE-C vs. NE-C: 0.61-fold, p < 0.01), although the viability was still lower than that in the NE-C cells (HE-S vs. NE-C: 0.85-fold) and Se-Met-treated cells (HE-S vs. NE-S: 0.81-fold). Se-Met relieved heat-induced decrease in testosterone level (HE-S vs. HE-C: 1.84-fold, p < 0.05) and HSD3B expression (HE-S vs. HE-C: 1.67-fold, p < 0.05). Se-Met alleviated heat-induced increase in Bcl2-associated protein X (BAX) expression (HE-C vs. HE-S: 2.4-fold, p < 0.05), and decrease in B-cell lymphoma-2 (BCL2) expression (HE-S vs. HE-C: 2.62-fold, p < 0.05), resulting in increased BCL2/BAX ratio in the HE-S cells (HE-S vs. HE-C: 5.24-fold, p < 0.05). Furthermore, Se-Met alleviated heat-induced activation of p-p38MAPK/p38MAPK (HE-C vs. HE-S: 1.79-fold, p < 0.05) and p-HSPB1/HSPB1 (HE-C vs. HE-S: 2.72-fold, p < 0.05). In conclusion, p38MAPK/HSPB1 might be involved in Se-Met-mediated alleviation of heat-induced cell apoptosis, cell viability and testosterone secretion impairments in sheep LCs.

To evaluate the effects of dietary quercetin on growth performance, blood parameters and testicular development of Hu sheep in summer, 96 lambs (25.51 +/- 0.23 kg) aged 4 months were randomly assigned to two groups (Con and Que, n=48), fed a basal diet supplemented without (Con) or with 3.75 g/d (Que) quercetin, respectively. Lambs were fed for 5 weeks including a pretrial adaption of 1 week. Feed intake and body weight were recorded, blood samples and testicular tissues were collected to detect the biological indexes, immunoglobulins, antioxidants, hormones, testicular spermatogenic epithelium and steroidogenic proteins. Results showed that quercetin supplement decreased feed intake very significantly (P<0.01) without affecting average daily gain; increased serum NEFA level significantly (P<0.05) without affecting concentrations of Glu, TC, TG, HDL, or LDL. Quercetin supplement increased the activity of serum GPX, and concentrations of plasma IgA, IgM, and IgG very significantly (P<0.01); However, it did not significantly affect T-AOC level or SOD activity. Quercetin supplement reduced T3 concentration numerically (P=0.050) without changing plasma T4 or cortisol concentration (P>0.05). Testicular spermatogenic cells in the Que group were arranged in a more organized manner with a thicker epithelium (P<0.01), and plasma testosterone increased numerically compared with that in the Con group (P=0.071). WB analysis showed that quercetin supplement increased testicular HSD3B expression very significantly (P<0.01) without affecting StAR, P450scc, or CYP17 expressions. In conclusion, dietary supplement of quercetin at 3.75 g/d in summer can decrease food intake without affecting the body weight, promote lipid metabolism, antioxidant capacity, immune response and improve the structure of testicular spermatogenic epithelium in Hu sheep, which provide evidences for further application of quercetin in sheep production.

This study aimed to investigate the role of NR4A1 in forskolin (FSK)-induced granulosa cell (GC) differentiation and PGF2α-induced granulosa-lutein cell (GLC) regression. For experiment 1, primary porcine GCs were pre-cultured for 6 d before induced-differentiation by FSK with or without siNR4A1, and changes in GC proliferation, lipid droplets (LDs), and P4 level were detected. For experiment 2, the GLC model was established by FSK as in experiment 1, and then PGF2α was utilized to induce GLC regression with or without siNR4A1, changes in P4 secretion, apoptosis proteins, and associated signaling pathway members were detected. Results showed that in experiment 1, FSK up-regulated NR4A1 expression during GC differentiation and decreased GC proliferation activity, which was reversed by siNR4A1. siNR4A1 inhibited the FSK-induced decreases in Cyclin B1/D1 and CDK1/2 mRNA abundances, and increases in P21/P27 mRNA abundances, and FSK-induced LD accumulation. FSK up-regulated P4 secretion and StAR, CYP11A1 and HSD3B expression, decreased CYP19A1 expression, which were reversed by siNR4A1 except for StAR expression. In experiment 2, PGF2α induced NR4A1 expression and reduced GLC viability, which were reversed by siNR4A1. Compared with PGF2α group, the levels of P4 secretion and StAR expression were higher in PGF2α+siNR4A1 group, while CYP11A1 and HSD3B expressions held at low levels. siNR4A1 inhibited PGF2α-induced expression of apoptosis proteins (caspase3, Bax, Fas, TNFa), ATF3, and phosphorylated MAPKs (ERK1/2, P38, JNK). In summary, NR4A1 is involved in regulating porcine GC differentiation and GLC regression as well as the changes in cell proliferation, apoptosis, steroidogenesis, and MAPK pathways, which provide a theoretical basis for further understanding of the mechanism of porcine luteal formation and regression.

[目的]本文旨在通过对猪卵泡颗粒细胞进行转录组测序了解卵泡闭锁相关基因的表达谱并确定闭锁卵泡的潜在分子标记物.[方法]通过HE染色对卵泡及颗粒细胞进行形态学观察;抽取健康卵泡和晚期闭锁卵泡的颗粒细胞,进行转录组测序分析,最后采用实时荧光定量PCR检测差异表达基因的相对表达水平.[结果]转录组数据表明,与闭锁卵泡颗粒细胞相比,健康卵泡颗粒细胞中有771个差异基因,其中204个为上调基因,567个为下调基因;GO功能富集分析显示,差异基因显著富集到320个生物进程中,主要与细胞增殖和血管生成有关;KEGG通路分析表明,差异基因富集到48条信号通路中,主要参与氧化应激、激素合成和能量代谢等方面的信号通路;实时荧光定量PCR结果表明,CYP1B1、PDE10A、NUPR1、NOX4、LDLR、LHCGR和CYP19A1表达趋势与测序结果一致.[结论]氧化应激、血管生成、能量代谢以及激素合成等多方面因素综合调控猪卵巢中颗粒细胞的凋亡,继而影响卵泡闭锁现象的产生.

Context Bone morphogenetic proteins (BMPs) play an important role in the uteri. Repulsive guidance molecule b (RGMb; a.k.a. Dragon) has been confirmed as the coreceptor of BMPs to function through drosophila mothers against decapentaplegic protein (Smads) and mitogen-activated protein kinases (MAPK) pathways. We hypothesise that RGMb regulates the uterine function through the Smads and MAPK pathways. Aims This study aimed to investigate the expression of RGMb in goat uteri and the potential role of RGMb in the endometrial epithelial cells (EECs). Methods The localisation of RGMb in goat uterine tissues was detected by immunohistochemistry (IHC), EECs were isolated and transfected with siRNA to investigate the role of RGMb in proliferation, and apoptosis. The expression levels of Smads and MAPK members was measured by western blot (WB) and real-time PCR (RT-PCR). Key results IHC showed that RGMb was localised in goat endometrial luminal cells, glandular epithelial cells, and circular muscle fibres, but not in stromal cells. RT-PCR results showed that treatment with RGMb siRNA suppressed the expressions of proliferation-related genes cyclin D1 (CCND1, P = 0.0291), cyclin-dependent kinase 2 (CDK2 P = 0.0107), and proliferating cell nuclear antigen (PCNA, P = 0.0508), leading to the reduced viability of EECs (P = 0.0010). WB results showed that the expression ratio of cleaved-caspase 3/caspase 3 (P = 0.0013) was markedly increased after RGMb siRNA transfection. Likewise, the level of phospho-extracellular signal-regulated kinase 1/2 (p-ERK1/2, P = 0.0068) and p-Smad1/5/8 (P = 0.0011) decreased significantly, while there were no appreciable differences in the level of p-P38 MAPK expression (P > 0.05). Conclusions RGMb might participate in the regulation of cell proliferation and apoptosis through Smads and ERK signalling pathways in goat EECs. Implications RGMb is involved in regulating the proliferation and apoptosis in goat endometrial epithelial cells.

[目的]本研究旨在探究夏季日粮添加酵母硒对湖羊血液指标和睾丸发育的影响,解析酵母硒影响睾丸功能的潜在分子机制.[方法]将96只4月龄雄性湖羊按体重相近原则分为对照组(Con,基础日粮)和酵母硒组(Se,基础日粮+酵母硒0.4 g·kg-1).预试期1周,正试期4周,记录体重变化;收集血液检测生化、抗氧化与免疫相关指标;采集睾丸组织,测质量和大小及其硒浓度,HE染色观察组织学形态(HE),转录组测序(RNA-seq)分析差异基因.[结果]Se组平均日增重显著高于Con组(P<0.05),2组间睾丸的大小、质量及系数无显著差异(P>0.05),但Se组睾丸中硒浓度极显著高于Con组(P<0.01).与Con组相比,Se组血清游离脂肪酸(NEFA)浓度显著升高(P<0.05),血浆IgA、IgG和IgM浓度极显著升高(P<0.01),血清和睾丸组织中GSH-Px活性极显著升高(P<0.01).HE结果显示生精小管中有精子出现,Se组生精细胞排列更紧密且生精上皮厚度极显著增加(P<0.01).RNA-seq分析鉴定出2321个差异表达基因,包含671个上调基因和1650个下调基因,通过GO和KEGG功能分析,差异基因主要富集在电子转移链、呼吸链复合体、呼吸链、线粒体膜部分、NADH脱氢酶(泛醌)活性、NADH脱氢酶(醌)活性和产热等通路.[结论]夏季日粮添加酵母硒可促进湖羊的脂质代谢,改善其抗氧化能力和免疫功能,改变湖羊睾丸发育相关基因的表达,为进一步阐明酵母硒缓解夏季高温对湖羊睾丸损伤的分子机制提供科学依据.

猪附红细胞体寄生于红细胞表面或游离于血浆、组织液及脑眷液中,引起猪发热、溶血性贫血和黄疽等主要症状.本试验用猪附红细胞体抗原和弗氏佐剂乳化制备免疫原,免疫试验兔,并通过酶联免疫吸附试验(ELISA)检测血清的效价,当测得血清的OD值与阴性对照组的血清值之比(P/N)分别为8.02(P/N＞ 2.1)、7.16(P/N＞ 2.1),采集兔血分离阳性血清.本试验为进一步研究猪附红细胞体的粘附阻抑以及该病的诊断、治疗提供了试验材料,并探索出了兔抗猪附红细胞体阳性血清的制备方法.

To explore the possibility of heterologous blood transfusion to dog after removing surface antigen of pig red blood cells,the osmotic fragility of pig red blood cells modified by a galactosidase glycosidase and mPEG-SC in vitro was detected,and experiment of matching of blood was done.The clinical indicators of dog were observed,and blood and urine routine was detected and analyzed continuously before and after blood transfusion.The results showed that the modification reduced erythrocyte osmotic fragility and the blood did not appear to agglutination in vitro.The dog had no obvious clinical abnormal reactions after transfusion,and its indexes of blood and urine were normal.Conclusion,It is feasible that pig red blood cells modified are heterologous transfused to dog firstly,and could be used for veterinary clinic as general all purpose blood.