IntroductionThe number of litters is an important reproductive trait, which is one of the main indicators reflecting the production level and economic benefit of the pig farm. As an important reproductive organ of female mammals, the ovary is controlled by a complex transcription network of coding and non-coding genes to undergo a series of biological processes during each estrus cycle, thereby regulating the reproductive capacity of the sow. However, these molecular regulation mechanisms affecting sow litter size are still unclear.MethodsRegarding the non-coding molecular regulatory mechanisms in ovarian function with smaller and larger litter size (SLS and LLS), we investigated the expression profile of lncRNA in pig SLS and LLS samples. Total RNAs from porcine ovaries were used to construct libraries using Ribo-Zero RNA sequencing method.ResultsHere we profiled the expression of lncRNA in porcine ovaries with SLS and LLS, and identified a total of 3,556 lncRNA candidates, of which 96 were upregulated lncRNA and 206 were downregulated lncRNA when comparing LLS to SLS. In addition, a competitive endogenous RNA (ceRNA) network was constructed, and it was found that lncRNAs LOC100513133 and LOC102168075 may serve as ceRNAs containing potential binding sites for miR-26b, let-7g and miR-125b.DiscussionThese results demonstrate that lncRNAs may play roles in modulating porcine litter size.

Reproductive disorders are a concern in the pig industry. Successful gestation processes are closely related to a suitable endometrial microenvironment, and the physiological mechanisms leading to failed pregnancy during the peri‐implantation period remain unclear. We constructed single‐cell transcriptome profiles of peri‐implantation embryo loss and successful gestation endometrial tissues and identified 22 cell subpopulations, with epithelial and stromal cells being the predominant endometrial cell types. The two tissues showed marked differences in cell type composition, especially among epithelial cell subpopulations. We also observed functional differences between epithelial and stromal cells in tissues from embryonic loss and successful gestation, as well as the expression levels and differentiation trajectories of genes associated with embryo attachment and endometrial receptivity in epithelial and stromal cells. The results of cell communication interactions analysis showed that ciliated cells were more active in endometrial tissue with embryo loss, and there were differences in the types of endometrial cells with major roles in embryo loss and embryo implantation successful tissues for bone morphogenic protein, insulin‐like growth factor, and transforming growth factor‐β signaling networks associated with embryo implantation. In addition, we compared the functional differences in immune cells between the two tissue types and the expression levels of genes related to the inflammatory microenvironment. Overall, the present study revealed the molecular features of endometrial cell transcription in embryo‐lost endometrial tissues, providing deeper insights into the endometrial microenvironment of reproductive disorders, which may inform the etiological, diagnostic, and therapeutic studies of reproductive disorders.

Low sperm motility is a significant contributor to male infertility. beta-defensins have been implicated in host defence and the acquisition of sperm motility; however, the regulatory mechanisms governing their gene expression patterns and functions remain poorly understood. In this study, we performed single-cell RNA and spatial transcriptome sequencing to investigate the cellular composition of testicular and epididymal tissues and examined their gene expression characteristics. In the epididymis, we found that epididymal epithelial cells display a region specificity of gene expression in different epididymal segments, including the beta-defensin family genes. In particular, Defb15, Defb18, Defb20, Defb25 and Defb48 are specific to the caput; Defb22, Defb23 and Defb26 to the corpus; Defb2 and Defb9 to the cauda of the epididymis. To confirm this, we performed mRNA fluorescence in situ hybridisation (FISH) targeting certain exon region of beta-defensin genes, and found some of their expression matched the sequencing results and displayed a close connection with epididimosome marker gene Cd63. In addition, we paid attention to the Sertoli cells and Leydig cells in the testis, along with fibroblasts and smooth muscle cells in the epididymis, by demonstrating their gene expression profile and spatial information. Our study provides a single-cell and spatial landscape for analysing the gene expression characteristics of testicular and epididymal environments and has important implications for the study of spermatogenesis and sperm maturation.

生猪产业是我国畜牧业中的主导产业,是满足人们日益增长的肉食需求的主要来源,在生猪产业中,种猪育种重要性占 40%以上.该文以陕西正能集团育种工作的实践,简要阐述了种猪档案管理、种猪性能测定、表型选择、联合育种、分子育种和大数据技术等方法的基本流程;通过组建育种核心群,选择目标性状,采用配合力测定等技术的应用,使正能集团的长白种猪和大白种猪各项性能稳步提升,尤其在繁殖性能上取得了显著的进展,以期为其他种猪企业提供可参考的案例.

精液品质是公畜选种的重要指标,也是影响公畜遗传潜能发挥的重要因素,对于畜禽生产具有十分重要的意义.肠道微生物作为机体最大的微生态系统,参与物质和能量代谢,对机体健康有重要的影响.研究发现肠道微生物不仅与肥胖、炎症、癌症等生理病理过程息息相关,而且可以调控雄性动物精液品质和繁殖性能.本文就肠道微生物对公畜精液品质及其繁殖性能影响的相关研究进展进行综述,以期为提高公畜精液品质、促进优良基因传播提供一些思路.

母猪繁殖性能对我国生猪产业的持续发展极其重要,其主要受到营养、内分泌激素、饲养管理等因素影响.肠道微生物作为宿主内最大的微生物系统,能通过脑-肠轴等多个代谢途径与宿主相互作用,广泛参与宿主机体活动.目前,已有研究发现肠道微生物与母猪生殖激素、血清代谢物以及炎症反应等之间的相关性,本文对肠道微生物影响母猪繁殖性能的相关研究进行综论,为后续通过肠道微生物提高母猪健康和繁殖性能、促进生猪产业发展提供理论基础.

Intramuscular fat content is an important factor that determines meat quality in pigs. In recent years, epigenetic regulation has increasingly studied the physiological model of intramuscular fat. Although long noncoding RNAs (lncRNAs) play essential roles in various biological processes, their role in intramuscular fat deposition in pigs remains largely unknown. In this study, intramuscular preadipocytes in the longissimus dorsi and semitendinosus of Large White pigs were isolated and induced into adipogenic differentiation in vitro. High-throughput RNA-seq was carried out to estimate the expression of lncRNAs at 0, 2 and 8 days post-differentiation. At this stage, 2135 lncRNAs were identified. KEGG analysis showed that the differentially expressed lncRNAs were common in pathways involved with adipogenesis and lipid metabolism. lnc_000368 was found to gradually increase during the adipogenic process. Reverse-transcription quantitative polymerase chain reaction and a western blot revealed that the knockdown of lnc_000368 significantly repressed the expression of adipogenic genes and lipolytic genes. As a result, lipid accumulation in porcine intramuscular adipocytes was impaired by the silencing of lnc_000368. Overall, our study identified a genome-wide lncRNA profile related to porcine intramuscular fat deposition, and the results suggest that lnc_000368 is a potential target gene that might be targeted in pig breeding in the future.

Abstract Spermatozoa produced in the testes acquire motility and maturation processes in the epididymis, but little is known about the mechanism of sperm development in the testicular and epididymal environment. In this study, we performed single-cell RNA and spatial transcriptome sequencing to study the mechanism of mouse spermatogenesis. We investigated the cell compositions of testicular and epididymal tissues and examined their gene expression characteristics. In the epididymis, we found that epididymal epithelial cells display a specific gene expression profile in different regions of the epididymis. Therefore, we divided the entire epididymis into 12 segments according to spot cluster and structural characteristics and investigated their gene expression profiles. Moreover, we investigated the detailed expression characteristics of β-defensin family genes in the mouse epididymis. In addition, we also analyzed the gene expression characteristics and spatial information of stromal cells associated with spermatogenesis, including Sertoli cells and Leydig cells in the testis and fibroblasts and smooth muscle cells in the epididymis. Our study provides a single-cell and spatial landscape to analyze the gene expression characteristics of the testicular and epididymal environments in which spermatogenesis and maturation occur and has important implications for the study of spermatogenesis and maturation.

【Objective】 This study was aimed to investigate the effects of Clostridium butyrate（C.butyricum） and sodium butyrate on semen quality and testicular tissue of mice stimulated by high fat diet, so as to provide experimental basis and theoretical support for the treatment of male reproductive diseases.【Method】 In experiment 1,32 8-week-old C57BL/6 male mice were randomly divided into 4 groups.The mice in control group（NC） were fed a basal diet, in high fat diet group（HFD）, C. butyricum treatment group（HFD+C.butyricum） and sodium butyrate treatment group（HFD+Sodium butyrate） were fed high fat diet containing 60% fat.HFD+C.butyricum group was gavaged with 1×10~8 CFU/kg C.butyricum,while HFD+Sodium butyrate group was gavaged with 300 mg/kg sodium butyrate, once a day.In experiment 2,16 8-week-old C57BL/6 male mice were randomly divided into two groups.The normal diet of control group was supplemented with normal saline, and the experiment group was supplemented with C.butyricum solution 1×10~8CFU/kg by gavage, once a day.After 12 weeks, the animals were sacrificed for sampling.The semen quality was detected by CASA,and the pathological changes of testicular tissue were observed by HE staining.The total antioxidant capacity（T-AOC）,the activities of superoxide dismutase（SOD） and glutathione peroxidase（GSH-Px）,the content of malondialdehyde（MDA） in testicular tissue and serum testosterone（T）, and the activities of alkaline phosphatase（AKP）,acid phosphatase（ACP）,lactate dehydrogenase（LDH） and gamma glutamyltransferase（γ-GT） in testicular tissue were detected by corresponding kits.【Result】 In experiment 1,compared with high-fat group, the body weight of mice in C.butyricum treatment group was extremely significantly decreased（P＜0.01）,the sperm motility, the number of A+B grade sperm, sperm linear motility and the velocity of average path of mice were significantly increased（P＜0.05）,and the straight-line velocity of mice was extremely significantly increased（P＜0.01）.The activity of AKP in testicular tissue of mice was significantly increased（P＜0.05）.Compared with high-fat group, the body weight of mice in sodium butyrate treatment group was extremely significantly decreased（P＜0.01）,the testis index, sperm linear motility, sperm forward motility, straight-line velocity and swing of sperm were significantly increased（P＜0.05）,and the sperm motility, the number of A+B grade sperm, the velocity of average path and the activity of γ-GT in testicular tissue were extremely significantly increased（P＜0.01）.In addition, sodium butyrate significantly increased the content of serum testosterone and the activity of ACP（P＜0.05）,and significantly decreased the content of MDA in testicular tissue of mice（P＜0.05）.The pathological section of testicular tissue showed that there was no significant difference in testicular tissue morphology of mice in each group.In experiment 2,compared with control group, the testicular coefficient of mice treated with C.butyricum were extremely significantly increased（P＜0.01）,and the level of serum testosterone was significantly increased（P＜0.05）,but there were no significant change in semen quality and antioxidant indexes of testicular tissue（P＞0.05）.【Conclusion】 C.butyricum and sodium butyrate could significantly increase the content of serum testosterone in mice, improve the antioxidant capacity of testicular tissue, improve the quality and quantity of sperm, protect the structure of testicular tissue, and effectively protect the reproductive system injury caused by obesity in male mice.

为研究复合益生菌发酵饲料对大白公猪精液质量的影响,选择12头大白种公猪,并随机分成两组,开展了复合益生菌发酵饲料饲喂试验,测定公猪血液生理生化指标、免疫性能和精液质量等指标.结果表明,饲喂发酵饲料60 d后,试验组公猪血清尿素氮含量、中间细胞数量以及中间细胞比例明显下降(P<0.05),血液白细胞数和平均红细胞体积显著升高(P<0.05),IgA和IgM含量呈现出上升的趋势(P>0.05).与对照组相比,试验组公猪的精子活力与密度极显著提高(P<0.01),顶体和质膜完整性以及线粒体膜电位活性显著提高(P<0.05).试验第120 d,试验组公猪精液中丙二醛的含量和活性氧水平极显著降低(P<0.01),精浆中果糖含量极显著提高(P<0.01).综上所述,复合益生菌发酵饲料能够显著提高精子密度、活率、质膜和顶体完整性以及精浆果糖含量,改善大白种公猪的免疫力,降低精液的氧化损伤,进而改善精液品质.

The number of live births in a litter is an important reproductive trait, and is one of the main indicators which reflect the production level and economic benefit of a pig farm. The ovary is an important reproductive organ of the sow, and it undergoes a series of biological processes during each estrous cycle. A complex transcriptional network containing coding and non-coding RNAs in the ovary closely regulates the reproductive capability of sows. However, the molecular regulation mechanisms affecting sow litter size are still unclear. We investigated the expression profiles of microRNAs (miRNAs) in porcine ovaries from sows with smaller than average litter sizes (SLS) and those with larger litter sizes (LLS). In total, 411 miRNAs were identified, and of these 17 were significantly down-regulated and 16 miRNAs were up-regulated when comparing sows with LLS and SLS, respectively. We further characterized the role of miR-183 which was one of the most up-regulated miRNAs. CCK-8, EdU incorporation and western blotting assays demonstrated that miR-183 promoted the proliferation of granulosa cells (GCs) in pig ovaries. Moreover, miR-183 inhibited the synthesis of estradiol in GCs and promoted the synthesis of progesterone. These results will help in gaining understanding of the role of miRNAs in regulating porcine litter size.

Homeostasis of gut microbiota is a critical contributor to growth and health in weaned piglets. Fish oil is widely reported to benefit health of mammals including preventing intestinal dysfunction, yet its protective effect during suckling-to-weaning transition in piglets remains undetermined. Low (30 g/d) and high (60 g/d) doses of n-3-rich fish oil were supplemented in sows from late gestation to lactation. Serum indicators and gut microbiota were determined to evaluate the effects of maternal fish oil on growth performance, immunity and diarrhea of piglets. DHA and EPA in the colostrum as well as serum of suckling and 1-week post-wean piglets were significantly and linearly increased by maternal supplementation of fish oil (P < 0.05). IGF1 and T3 in nursing and weaned piglets were significantly elevated by maternal fish oil (P < 0.05), and the increase of IGF1 was concerning the dosage of fish oil. Colostrum IgG, plasma IgG, IgM in suckling piglets, IgG, IgM and IgA in weaned piglets were significantly increase as maternal replenishment of fish oil increased (P < 0.05). Additionally, cortisol was significantly reduced in weaned pigs (P < 0.05), regardless of dosage. 16S rRNA sequencing revealed that α-diversity of fecal microbiota in nursery piglets, and fecal Lactobacillus genus, positively correlated with post-weaning IgA, was significantly increased by high dosage. Collectively, maternal fish oil during late pregnancy and lactation significantly promoted growth, enhanced immunity, and reduced post-weaning diarrhea in piglets, therefore facilitated suckling-to-weaning transition in piglets, which may be partially due to the altered gut microbial community.

MicroRNAs (miRNAs) are emerging as an important regulator during adipose development. Previous studies have revealed that miR-370 is related to lipid metabolic homeostasis, however, its roles in lipid accumulation remain to be established. In this study, we demonstrated that ssc-miR-370 was highly conserved across domestic animals, and relatively enriched in adipose tissues and skeletal muscles. Overexpression of ssc-miR-370 significantly promoted the proliferation of porcine preadipocytes through facilitating G1/S phase transition. Meanwhile, ssc-miR-370 mimics dramatically suppressed adipogenic differentiation, indicated by reduced triglyceride deposition as well as downregulated PPARγ and aP2 expressions. Furthermore, ssc-miR-370 was demonstrated to repress FoxO1 expression via directly targeting FoxO1 3'-UTR using dual luciferase activity assay. Our data evaluates miR-370 as a novel adipogenic modulator, which may be a potential target to reduce backfat thickness in pigs and fight obesity in humans.

试验旨在探究母猪日粮分别添加亚麻油和鱼油对母猪和仔猪血清生化指标、免疫指标和生产性能的影响.选取27头体况相近、2胎次的长白母猪随机分为对照组、2％亚麻油组和2％鱼油组,每组9个重复,每个重复1头母猪,试验期从妊娠第85天开始至断奶结束.结果表明:亚麻油和鱼油对母猪生产性能和血清生化指标无显著影响,亚麻油可提高仔猪血清中总蛋白含量(P<0.05),鱼油可提高仔猪血清中谷草转氨酶/谷丙转氨酶比例(P<0.05);亚麻油和鱼油均提高了母猪初乳血清和仔猪血清中免疫球蛋白和炎症因子水平(P<0.05),对母猪和仔猪血清中氧化指标均无显著影响.综上,母猪妊娠后期日粮添加亚麻油和鱼油可改善仔猪血清生化指标,提高免疫球蛋白和炎症因子水平.

miRNAs are a small class of noncoding RNAs that perform biological functions by regulating the stability or translation of target genes in various biological processes. This study illustrated the role of miR-10a-5p, which is relatively enriched in adipose tissues, using primary mouse preadipocytes as model. With elevated miR-10a-5p expression, the proliferative ability of mouse preadipocytes was significantly enhanced, indicated by increased EdU+ cells and G1/S transition, accompanied by upregulated Cyclin B, Cyclin D and PCNA and downregulated p21 and p27. Meanwhile, the adipogenic differentiation was significantly attenuated by elevated miR-10a-5p, supported by Oil Red O staining and suppressed PPARγ and aP2 expression. Furthermore, Map2k6 and Fasn were predicted to be the target genes of miR-10a-5p in silico, and dual luciferase reporter assay confirmed the direct targeting effects. Western blot analysis results showed that miR-10a-5p specially reduced Map2k6 expression at the proliferative stage without affecting Fasn expression, while significantly restrained Fasn expression with unchanged Map2k6 expression during adipogenic differentiation. Taken together, these results revealed a potential role of miR-10a-5p in adipogenesis and in the treatment of obesity.

Excess intramyocellular lipids are often accompanied by muscle insulin resistance (IR) and type 2 diabetes. The mechanism of the formation of intramyocellular lipids is unclear yet. In this study, we optimized the cellular model of intramyocellular lipids from differentiated C2C12 cells and identified that the expression of insulin-like growth factor-binding protein 5 (IGFBP5) is diminished in this process. Then, we added exogenous recombinant IGFBP5 during myocyte triglyceride (TAG) formation and found decreased lipids accumulation. In addition, IGFBP5 could promote lipolysis when added to the cellular model after the formation of intramyocellular lipids. Moreover, IGFBP5 could enhance myocyte insulin sensitivity by inhibiting the expression of the thioredoxin-interacting protein (TXNIP) and arrestin domain-containing 4 (ARRDC4), which are a negative regulator of insulin signaling in both cases. Meanwhile, IGFBP5 also inhibited the expression of glycerol-3-phosphate acyltransferase (GPAM) and diglyceride acyltransferase 2 (DGAT2), which were involved in TAG synthesis from a fatty acid. IGFBP5 also reduced TAG storage by promoting lipolysis. Therefore, IGFBP5 may play a role in the excess accumulation of lipid in muscle cells of diabetic patients and serve as a reference for further research and treatment of muscle IR and diabetes.

为探究miR-106a-5p对成肌分化的作用及潜在调控靶点,在线预测miR-106a-5p潜在靶基因,通过RT-qPCR及Western blot检测miR-106a-5p对靶基因的调控作用,利用双荧光素酶报告系统验证miR-106a-5p与靶基因之间的互作,并以C2C12细胞系为实验模型,采用过表达技术,在细胞形态学水平初步探索其对成肌分化的作用.在线预测发现E2F3和SP-1可能是miR-106a-5p的潜在靶基因;RT-qPCR分析表明,miR-106a-5p与E2F3和SP-1的表达模式相反;Western blots结果发现,miR-106a-5p抑制E2F3和SP-1蛋白翻译;双荧光素酶报告载体系统表明,E2F3和SP-1是miR-106a-5p调控成肌分化的靶基因;此外,超表达miR-106a-5p抑制C2C12细胞分化,肌管数目显著减少.结果 表明,E2F3和SP-1是miR-106a-5p抑制C2C12细胞成肌分化的靶基因.

Circular RNAs (circRNAs) have been emerging as an important regulator in mammalian reproduction via acting as miRNA sponges. However, the circRNAs in porcine ovaries related with litter size remains largely unknown. In this study, porcine ovaries with smaller or larger litter size (LLS) were subjected to high-throughput RNA sequencing. In total, 38,722 circRNAs were identified, of which 1,291 circRNAs were commonly expressed in all samples. There were 56 circRNAs significantly down-regulated and 54 circRNAs up-regulated in LLS pig (|log2 (fold change) | > 1, FDR < 0.05). Bioinformatics predicted that most of circRNAs harbored miRNA binding sites, and the expression patterns of circRNAs and their putative binding miRNAs were validated by qPCR. Moreover, the expression of circ-TCP11/miR-183 was significantly reversely correlated and their direct interaction was confirmed by dual-luciferase assay. Our study indicates that circRNAs may play potential effects on modulating porcine litter size.

试验选择出生日龄相近、健康、体重约25 kg的太蕨猪（太湖猪×蕨麻猪）和巴蕨猪（巴克夏×蕨麻猪）去势公猪各20头,测定其体尺、生长性能、胴体品质及肉质,以筛选出适合生产需要的杂交组合。结果表明,在2月龄、4月龄、6月龄巴蕨猪体高和体长指标均显著高于太蕨猪,胸围、腹围、胸深、管围两品种间差异不显著;与太蕨猪相比,巴蕨猪日增重显著提高18.64%,料重比显著降低11.40%,眼肌面积极显著高71.66%,瘦肉率显著高12.68个百分点,剪切力显著降低18.71%;但是太蕨猪肉色优于巴蕨猪。综合比较发现,巴克夏猪与蕨麻猪杂交在生长性能和肉质性状等指标的改良效果均优于太湖猪与蕨麻猪杂交,本试验条件下巴克夏猪与蕨麻猪的杂交组合更适合实际生产需要。

为鉴定、验证、评价分光光度测量法在试管凝集试验结果判定中的实用价值,按照《动物布鲁氏菌病诊断技术》(GB/T 18646-2002)试管凝集试验操作方法检测72份已知凝集效价血清,比较分光光度测量法和比浊法判定结果差异.结果显示,分光光度测量法和比浊法判定72份血清效价与已知效价符合率分别为88.89％(64/72)、83.33％(60/72)(P>0.05),判定结果一致率为94.44％,Kappa值0.77>0.75,但分光光度测量法判定效价与已知效价符合率高于比浊法,且对试验结果的判定更为客观、准确,因此,分光光度测量法判定试管凝集试验结果适宜在布鲁菌病监测中推广应用.