In recent years, the meat and dairy value of buffaloes has become a major concern in buffalo breeding, and the improvement of buffalo beef quality is key to protecting buffalo germplasm resources and solving the problem of beef supply. MiRNAs play a significant role in regulating muscle development. However, the precise mechanism by which they regulate the development of buffalo skeletal muscles remains largely unexplored. In this study, we examined miRNA expression profiles in buffalo myoblasts during the proliferation and differentiation stages. A total of 177 differentially expressed miRNAs were identified, out of which 88 were up-regulated and 89 down-regulated. We focused on a novel miRNA, named bbu-miR-493-5p, that was significantly differentially expressed during the proliferation and differentiation of buffalo myoblasts and highly expressed in muscle tissues. The RNA-FISH results showed that bbu-miR-493-5p was primarily located in the cytoplasm to encourage buffalo myoblasts' proliferation and differentiation. In conclusion, our study lays the groundwork for future research into the regulatory role of miRNAs in the growth of buffalo muscle.

MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate several pathway intermediates and affect the skeletal muscle development in mice, pigs, sheep, and cattle. However, to date, only a small number of miRNAs have been reported in the muscle development of goats. In this report, the longissimus dorsi transcripts of one- and ten-month-old goats were analyzed by sequencing RNAs and miRNAs. The results showed that the ten-month-old Longlin goats had 327 up- and 419 down-regulated differentially expressed genes (DEGs) compared with the one-month-old. In addition, 20 co-up-regulated and 55 co-down-regulated miRNAs involved in the muscle fiber hypertrophy of goats were identified in ten-month-old Longlin and Nubian goats compared with one-month-old. Five miRNA-mRNA pairs (chi-let-7b-3p-MIRLET7A, chi-miR193b-3p-MMP14, chi-miR-355-5p-DGAT2, novel_128-LOC102178119, novel_140-SOD3) involved in the goat skeletal muscle development were identified by miRNA-mRNA negative correlation network analysis. Our results provided new insight into the functional roles of goat muscle-associated miRNAs, allowing a deeper understanding of the transformation of miRNA roles during mammalian muscle development.

Spermatogenesis is a complex process that requires many regulatory mechanisms to form healthy sperm. Numerous studies have also proved that m6A methylation modification and lncRNA are essential for normal spermatogenesis. However, the mutual regulation of m6A methylation and lncRNA in spermatogenesis is still unclear. In this study, we knocked down METTL3 in GC-1spg cells and found that a reduction in METTL3 increased cell proliferation. Further, we examined the lncRNA expression profiles of normal spermatogonia and spermatogonia with knocked down METTL3. We detected 30,924 lncRNAs, of which 34 were up-regulated and 77 down-regulated. The results of the MeRIP-qPCR experiment showed that ENSMUST00000186472, MSTRG.8019.3 and ENSMUST00000202148 had m6A methylation sites and were regulated by METTL3. We constructed ceRNA networks for these 3 lncRNAs. And we identified that these 3 lncRNAs might act as miRNA sponges to regulate some genes related to spermatogenesis. This study focuses on exploring the regulatory mechanisms of m6A methylation on lncRNAs in spermatogonia and provides some epigenetic theories for subsequent studies on the expression mechanisms of lncRNAs.

Abstract Background Buffalo is a characteristic species in Guangxi, whose meat is nutrient-dense and good for the human body because of its polyunsaturated fatty acids and conjugated linoleic acid. CircRNA is a type of non-coding RNA contributing to the growth of skeletal muscle via molecular sponges, protein scaffolds, and coded short peptides. Results Our prior RNA-seq results presented that the expression of circCOPS8 in the longissimus muscle of buffalo and cattle was considerably different. According to the results of western blotting and qPCR (P < 0.05), circCOPS8 improved the differentiation and suppressed the proliferation of buffalo myoblasts. However, the outcomes of the mouse muscle injury model showed that circCOPS8 hindered mice from repairing their damaged muscles. Further study on the mechanism of RIP-qPCR proved that circCOPS8 could bind to IGF2BP3 (P < 0.05). Transcriptome sequencing along with the outcomes of qPCR further demonstrated that circCOPS8 may also prevent cell growth by activating tumor suppressor genes (P < 0.05). Conclusion CircCOPS8 inhibits cell proliferation by binding to IGF2BP3 and promoting ATR gene expression. This study described the function and mechanism of circCOPS8, setting the theoretical groundwork for research of circRNAs regulating buffalo muscle growth.

In cis-regulatory elements, enhancers and promoters with complex molecular interactions are used to coordinate gene transcription through physical proximity and chemical modifications. These processes subsequently influence the phenotypic characteristics of an organism. An in-depth exploration of enhancers and promoters can substantially enhance our understanding of gene regulatory networks, shedding new light on mammalian development, evolution and disease pathways. In this review, we provide a comprehensive overview of the intrinsic structural attributes, detection methodologies as well as the operational mechanisms of enhancers and promoters, coupled with the relevant novel and innovative investigative techniques used to explore their actions. We further elucidated the state-of-the-art research on the roles of enhancers and promoters in the realms of mammalian development, evolution and disease, and we conclude with forward-looking insights into prospective research avenues.

[目的]研究鸡血藤总黄酮对环磷酰胺所致小鼠肝损伤的保护作用.[方法]将60只昆明系小鼠随机分为6组,每组10只,试验周期为7 d.对照组小鼠试验全程给予生理盐水.环磷酰胺(CTX)组小鼠1～4 d灌胃给予生理盐水;5～7 d腹腔注射80 mg/(kg·BW)CTX,每天给药1次.鸡血藤总黄酮(TFSD)100组、CTX+TFSD25组、CTX+TFSD50组、CTX+TFSD100组小鼠1～7 d分别灌胃给予100、25、50、100 mg/(kg·BW)TFSD,每天给药1次;试验5～7 d除TFSD100组小鼠腹腔注射生理盐水外,其余试验组腹腔注射80 mg/(kg·BW)CTX,每天给药1次.试验结束后对各组小鼠进行眼球采血及剖检;检测肝组织中超氧化物歧化酶(SOD)、髓过氧化物酶(MPO)、黄嘌呤氧化酶(XOD)以及谷胱甘肽过氧化物酶(GSH-Px)活力,测定血清中谷草转氨酶(AST)、谷丙转氨酶(ALT)和碱性磷酸酶(ALP)活力,检测血清中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和白介素-1β(IL-1β)含量.[结果]环磷酰胺可显著(P<0.05)降低小鼠肝脏中SOD、GSH-Px以及血清中ALT活力,显著(P<0.05)升高肝脏中XOD与MPO活力、血清中ALP活力以及炎症细胞因子IL-1β和IL-6释放水平.25、50、100 mg/(kg·BW)的TFSD处理均可显著(P<0.05)抑制由CTX引起的SOD活力降低,25 mg/(kg·BW)的TFSD处理可显著(P<0.05)提高CTX处理小鼠的GSH-Px活力;50 mg/(kg·BW)的TFSD处理可显著(P<0.05)抑制由CTX引起的XOD和MPO活力升高,且可显著(P<0.05)降低CTX处理小鼠血清中的AST活力;25、50、100 mg/(kg·BW)的TFSD处理可显著(P<0.05)抑制由CTX引起的血清中ALP活力升高,同时降低ALT活力.25、50、100 mg/(kg·BW)的TFSD处理均可显著(P<0.05)抑制由CTX引起的IL-1β水平升高.[结论]鸡血藤总黄酮可通过调节小鼠炎症因子释放以及肝组织中氧化还原酶水平对环磷酰胺所致肝损伤提供保护,推荐剂量为25 mg/(kg·BW).

2-Mercaptoethanol (2-ME) is often used as an antioxidant to optimize culture systems for in vitro oocyte maturation in livestock. However, the relationship between 2-ME and autophagy has not yet been elucidated. In this study, we hypothesized that 2-ME can promote porcine oocyte maturation in vitro by maintaining autophagy homeostasis. To test this hypothesis, we explored the effects of 2-ME on the maturation of porcine oocytes exposed to an autophagy activator (rapamycin) or an autophagy inhibitor (3-methyladenine, i.e., 3-MA) in vitro. Rapamycin-induced autophagy over-activation significantly increased autophagy- and apoptosis-related gene expression, oxidative stress, apoptosis rates, abnormal mitochondrial redistribution, and significantly decreased oocyte first polar body extrusion (PBE) rates, spindle/chromosome integrity and developmental competence. 3-MA-mediated autophagy inhibition exerted similar effects on all these parameters except the expression of genes that promote autophagy and inhibit apoptosis. Importantly, 2-ME supplementation significantly attenuated the detrimental effects of rapamycin and 3-MA. Interestingly, we observed that 44 h of coincubation with rapamycin/3-MA and 2-ME restored autophagy homeostasis in vitro. In conclusion, our study confirmed that 2-ME promotes porcine oocyte maturation and embryo development in vitro by maintaining autophagy homeostasis and lays a foundation for further research on the underlying mechanism.

[目的]研究miR-495-3p对山羊骨骼肌细胞增殖分化的影响及其在不同组织中的表达情况,为探究miRNA在肌肉发育中的调控机制提供理论基础.[方法]利用生物信息学方法预测miR-495-3p的靶基因;实时荧光定量PCR检测miR-495-3p及其靶基因在心脏、肝脏、脾脏、肺脏、肾脏、背最长肌、腿肌等组织中的表达水平.构建 miR-495-3p 的过表达(miR-495-3p mimic、miR-495-3p mimic NC)及抑制物(miR-495-3p inhibitor、miR-495-3p inhibitor NC),在山羊骨骼肌细胞汇合度达60％～70％时进行转染,并用含2％马血清的培养基进行诱导分化,用CCK-8检测细胞增殖活力,实时荧光定量PCR检测过表达和干扰效率及增殖分化相关基因配对盒基因(paired-box 7,Pax7)、细胞蛋白周期 E(Cyclin E)、肌细胞生成素(myogenin,myoG)、生肌因子5(recombinant myogenic factor 5,Myf5)的表达;构建miR-495-3p靶基因的野生型和突变型载体并转染至293T细胞,用双荧光素酶测定miR-495-3p与其靶基因的结合情况.[结果]生物信息学预测结果表明,miR-495-3p的靶基因为心肌肌动蛋白α1(alpha cardiacactin 1,ACTC1),且miR-495-3p和ACTC1在1和10月龄山羊背最长肌中表达量均差异极显著(P＜0.01).miR-495-3p和ACTC1在心脏、肝脏、脾脏、肺脏、肾脏、背最长肌、腿肌6种组织中均有表达,在背最长肌中表达量较高.细胞转染结果表明,miR-495-3p mimic极显著促进miR-495-3p的表达(P＜0.01),miR-495-3p inhibitor 极显著抑制 miR-495-3p 的表达(P＜0.01),说明 miR-495-3p mimic 和 miR-495-3p inhibitor 可用于后续试验.实时荧光定量PCR结果表明,过表达miR-495-3p促进骨骼肌细胞分化相关标志基因MyoG、Myf5的表达,抑制miR-495-3p则降低MyoG、Myf5基因的表达;过表达、干扰miR-495-3p对Pax7、Cyclin E基因的表达及细胞增殖活力均无显著影响(P＞0.05).双荧光素酶报告结果表明,miR-495-3p与ACTC1基因存在靶向关系.[结论]miR-495-3p对山羊骨骼肌细胞的增殖无显著影响,但可通过靶向ACTC1基因促进山羊骨骼肌细胞分化.

Poultry production was long plagued by coccidiosis, and the development of alternative therapies will make practical sense. In this work, 2 battery experiments were designed. In battery experiment 1, the best effect of 7 anticoccidial herbs (Sophora japonica Linn, Citrus aurantium L, leaf of Acer palmatum, bark of Magnolia officinalis, fruit peel of Punica granatum L., Eclipta prostrata L., and Piper sarmentosum Roxb.) against Eimeria tenella infection of 21-day-old male Chinese Guangxi yellow-feathered chickens were screened out by clinic indexes (bloody feces scores, cecal lesion scores, oocysts output, relative weight gain rate, and survival rate). According to the results from battery experiment 1 and other literature research, we selected 2 monomers which were extracted from fruit peel of Punica granatum L. for further battery experiment 2 which were similar with battery experiment 1. Clinic results showed that Punicalagin had better anticoccidial effect than Ellagic acid. The anticoccidial mechanism exploration results of Elisa, antioxidant test, and pathological observation showed that Punicalagin reduced the cecal inflammation, improved the expression of immunoglobulin in cecal tissue, improved cecal integrity, and restored its REDOX state. Results of 16S rRNA sequencing analysis showed that Punicalagin also maintained the fecal flora health during E. tenella infection through insignificantly increasing the proportion of Lactobacillus and Faecalibacterium as well as significantly reducing the proportion of pathogenic bacteria, Escherichia-Shigella. RNA-Seq analysis results suggested that Punicalagin may play a role in controlling E. tenella infection by interaction with cytochrome P450 family enzymes. Overall, Punicalagin has promising potential as an alternative therapy for chicken Eimeria tenella infection.

Myogenesis is an essential process that can affect the yield and quality of beef. Transcriptional studies have shown that histone deacetylase 11 (HDAC11) was differentially expressed in muscle tissues of 6 and 18 month old Longlin cattle, but its role in the regulation of myogenesis remains unclear. This study aimed to determine the role of HDAC11 in the proliferation and differentiation of bovine muscle stem cells (MuSCs). HDAC11 promoted MuSC proliferation by activating Notch signaling and inhibited myoblast differentiation by reducing MyoD1 transcription. In addition, overexpression of HDAC11 inhibited the repair regeneration process of muscle in mice. HDAC11 was found to be a novel key target for the control of myogenesis, and this is a theoretical basis for the development of HDAC11-specific modulators as a new strategy to regulate myogenesis.

Buffalo meat is of good quality because it is lean and tender, and could bring significant cardiovascular benefits. The underlying difference in muscle development and meat quality is a complex and precisely orchestrated process which has been demonstrated to be regulated by long non-coding RNAs (lncRNAs). However, the regulatory role of lncRNAs in the growth and development of buffalo skeletal muscle is still unclear. In this study, the Ribo-Zero RNA-Seq method was used to explore the lncRNA expression profiles of buffalo myoblasts during the proliferation and differentiation phases. A specific set of 9,978 lncRNAs was found. By comparing the expression profiles of lncRNAs, it was found that there were 1,576 differentially expressed lncRNAs (DELs) during buffalo myoblast differentiation. Twelve DELs were chosen and subsequently verified in eight different buffalo tissues during fetal and adult stages by using qPCR. Gene11007 was found to be one of the most down-regulated lncRNAs during buffalo myoblasts differentiation and it was subsequently characterized. EdU, CCK-8, qPCR and western blotting assays showed that gene11007 promoted the proliferation of buffalo myoblasts but it had no effect on cell differentiation. Our research may enrich the genome annotations of buffalo and provide a new molecular target for the in-depth understanding of the regulation of lncRNAs in skeletal muscle.

[目的]通过对1和60日龄广西麻鸡腿肌进行转录组测序(RNA-Seq),筛选出广西麻鸡生长发育关键基因.[方法]选择1和60日龄健康母鸡各3只,分别采集腿肌组织,利用Illumina NovaseqTM 6000平台进行mRNA转录组测序,利用DESeq2软件进行差异表达基因的分析,并对差异表达基因进行GO功能和KEGG通路富集分析,进一步筛选出生长发育相关基因,并用实时荧光定量PCR验证转录组测序结果.[结果]转录组测序结果显示,6个样本分别获得34 296 198～41 647 642条clean reads.与1日龄腿肌相比,60日龄腿肌共获得2 304个差异表达基因,其中998个基因上调,1 306个基因下调.GO功能富集分析发现,共获得富集条目572条,其中生物过程381条,细胞组分70条,分子功能121条.KEGG通路富集分析发现,共获得34个显著富集的信号通路,其中心肌收缩、MAPK信号通路、紧密连接、肌动蛋白细胞骨架信号通路、心肌细胞中的肾上腺素能信号传导等与生长发育相关.通过GO功能和KEGG通路富集分析获得5个生长发育相关基因,分别为肌球蛋白重链10(MYH10)、肌球蛋白链15(MYH15)、成纤维细胞生长因子10(FGF10)、成纤维细胞生长因子16(FGF16)、肌肉生长抑制素(GDF8)基因,其中MYH10、MYH15、FGF10为上调差异表达基因,FGF16和GDF8为下调差异表达基因.所选差异表达基因实时荧光定量PCR结果与转录组测序的基因表达水平一致,说明转录组测序结果可靠.[结论]本试验通过对广西麻鸡1和60日龄2个不同生长阶段进行转录组测序分析,获得MYH10、MYH15、FGF10、FGF16、GDF8共5个与生长发育相关的关键基因,为后续进一步探讨广西麻鸡生长发育的分子调控机制提供参考.

为了探究小肌肉蛋白(Small Muscle Protein X-link,SMPX)在隆林山羊中的蛋白结构和组织表达水平,本研究克隆隆林山羊SMPX,并进行生物信息和组织表达谱分析.从NCBI查找山羊SMPX基因序列设计引物,根据TRlzol法提取1月龄隆林山羊心、肝、脾、肺、肾、背肌的总RNA,反转录成cDNA,并克隆SMPX,通过qRT-PCR分析SMPX在不同组织中的表达水平,进行生物信息学分析.SMPX基因编码区序列全长253 bp,共编码255个氨基酸,与NCBI上山羊编码区序列同源性达100％.SMPX蛋白的分子质量20.61 ku,等电点为5.29,是酸性蛋白,其在1～22个氨基酸处具有信号肽.SMPX有11个磷酸化位点,在隆林山羊不同组织中的表达量不一样,推测它可能在不同组织中发挥不同的生物学作用.

为寻找影响广西麻鸡不同生长时期,胸肌和腿肌组织间的差异基因和相关信号通路,以120日龄的广西雌性麻鸡胸肌和腿肌为研究对象,利用RNA-Seq技术对广西麻鸡不同生长时期胸肌和腿肌组织中差异表达基因进行筛选.转录组测序使用illumina NovaseqTM 6000测序,生物学重复3次,以P<0.05为标准筛选差异表达基因,结果发现,肌肉组织中有1953个差异基因,其中上调基因1415个,下调基因538个.通过对KEGG(P<0.05)通路分析发现,MAPK信号通路、粘着斑、PPAR信号通路和心肌细胞的肾上腺素能信号参与整个发育阶段;GO功能富集分析发现,在生物过程(BP),FHL、RBP7、MYBPC1、MUSTN1、TGF-β3、MY01C、IGF2和ADAM12对广西麻鸡的生长调控影响较大.

Buffalo holds an excellent potential for beef production, and circRNA plays an important role in regulating myogenesis. However, the regulatory mechanism of circRNAs during buffalo skeletal muscle development has not been fully explored. In this study, circRNA expression profiles during the proliferation and differentiation stages of buffalo myoblasts were analysed by RNA-seq. Here, a total of 3,142 circRNAs candidates were identified, and 110 of them were found to be differentially expressed in the proliferation and differentiation stages of buffalo myoblast libraries. We focused on a 347 nt circRNA subsequently named circCLTH. It consists of three exons and is expressed specifically in muscle tissues. It is a highly conserved non-coding RNA with about 95% homology to both the human and the mouse circRNAs. The results of cell experiments and RNA pull-down assays indicated that circCLTH may capture PLEC protein, promote the proliferation and differentiation of myoblasts as well as inhibit apoptosis. Overexpression of circCLTH in vivo suggests that circCLTH is involved in the stimulation of skeletal muscle regeneration. In conclusion, we identified a novel noncoding regulator, circCLTH, that promotes proliferation and differentiation of myoblasts and skeletal muscles.

The microenvironment in the seminiferous tubules of buffalo changes with age, which affects the self-renewal and growth of spermatogonial stem cells (SSCs) and the process of spermatogenesis, but the mechanism remains to be elucidated. RNA-seq was performed to compare the transcript profiles of pre-pubertal buffalo (PUB) and adult buffalo (ADU) seminiferous tubules. In total, 17,299 genes from PUB and ADU seminiferous tubules identified through RNA-seq, among which 12,271 were expressed in PUB and ADU seminiferous tubules, 4,027 were expressed in only ADU seminiferous tubules, and 956 were expressed in only PUB seminiferous tubules. Of the 17,299 genes, we identified 13,714 genes that had significant differences in expression levels between PUB and ADU through GO enrichment analysis. Among these genes, 5,342 were significantly upregulated and possibly related to the formation or identity of the surface antigen on SSCs during self-renewal; 7,832 genes were significantly downregulated, indicating that genes in PUB seminiferous tubules do not participate in the biological processes of sperm differentiation or formation in this phase compared with those in ADU seminiferous tubules. Subsequently, through the combination with KEGG analysis, we detected enrichment in a number of genes related to the development of spermatogonial stem cells, providing a reference for study of the development mechanism of buffalo spermatogonial stem cells in the future. In conclusion, our data provide detailed information on the mRNA transcriptomes in PUB and ADU seminiferous tubules, revealing the crucial factors involved in maintaining the microenvironment and providing a reference for further in vitro cultivation of SSCs.

雌性动物卵泡发育是一个动态的、受众多激素及调控因子直接或间接参与的复杂生物学过程,其中卵巢颗粒细胞的增值和分化能调节卵泡的生长和成熟,颗粒细胞的凋亡,影响卵泡闭锁.环氧合酶-2(cyclooxygenase 2,COX-2)在卵巢中主要在颗粒细胞中表达,可通过对下游代谢产物的调节,以不同的方式抑制颗粒细胞凋亡,参与PI3 K/AKT/NOTCH/Wnt信号通路的调节,从而作为排卵诱导因子诱导排卵.

为了降低植入式神经电极引起的炎性组织反应,通过层叠法构建可电刺激释放抗炎药物地塞米松的碳纳米管与聚吡咯双层导电生物膜(MWCNTs/Dex@PPy/Dex).并采用场发射扫描电镜(FESEM)、傅里叶变换红外光谱(FT-IR)、紫外分光光度法(UV)、循环伏安扫描(CV)和交流阻抗(EIS)对生物膜表面形貌、成分组成、药物释放特性和电化学性能进行考察.FESEM结果显示,MWCNTs/Dex@PPy/Dex生物膜呈现纤维状纳米结构,FT-IR光谱中含有与地塞米松类似的特征吸收峰.经过循环伏安电刺激释放地塞米松后,与PPy/Dex生物膜相比,MWCNTs/Dex@PPy/Dex导电生物膜阻抗更低,释放Dex总量更多,且释放时间更久.此外,PPy/Dex膜在205个循环发生脱落,而经过300个循环后,MWCNTs/Dex@PPy/Dex生物膜无明显肉眼可见变化,稳定性好,为提高电极的使用性能提供了思路.

近年来,国家出台多项政策推动草食动物产业发展,广西肉牛肉羊产业在自治区党委和政府的支持下也取得了明显成效.本文基于对广西全区14个市辖区牛羊养猪主要县(区)的调研,系统总结了广西肉牛肉羊产业的基本现状,包括广西牛羊养殖的规模、品种、经营模式、产业发展的区域优势、产业扶贫及方式、产业发展遇到的问题等情况,最后提出加快广西牛羊产业发展的对策,为下一步推动广西全区肉牛肉羊产业高质量发展提供参考.

目的 探讨发酵桑树全部或部分代替豆粕日粮对陆川肉猪血液生化指标及屠宰性能影响效果.方法 试验选取36头陆川育肥肉猪随机分为3组,每组12头,分别饲喂发酵桑树代替全部豆粕(试验A组)或50％豆粕(试验B组)与对照组(全价日粮),对各组猪生化指标和屠宰性能进行测定,试验期90 d.结果 饲喂添加不同水平发酵桑树日粮的试验日粮与对照组比较,试验A组血液中葡萄糖、胴体膘厚和脂率含量与试验B组、对照组比较显著降低,而猪瘦肉率显著提高.结论 陆川育肥猪饲喂发酵桑树代替全部豆粕日粮,可提高猪胴体瘦肉率.