Intramuscular fat (IMF) deposition drives premium beef value, but phenotypic selection is costly and slow. Although genome-wide studies have linked the KIRREL3 region to meat quality, whether intronic variants directly affect carcass traits and retain predictive power across physiological stages remains unproven. We genotyped two intronic SNPs (g.29756010C>T and g.29756149C>T) in Woking cattle, a high-quality beef breed developed in Jilin Province, China (n = 473; 226 and 206 heifers at 36 and 48 months, respectively, plus an independent validation cohort of n = 41), assessed for fatty acid composition, meat quality, and multi-tissue gene expression. Both loci were polymorphic and in Hardy–Weinberg equilibrium. At 36 months, TT individuals showed higher dressing percentage, rib thickness, and meat grade than CC at both loci. By 48 months, the 63-bp locus retained broader associations with carcass traits, whereas the 203-bp locus effects weakened. In the validation group, CC animals had higher IMF and muscle KIRREL3 expression than TT, yet received lower meat grades. Five fatty acids differed significantly among genotypes after Benjamini–Hochberg correction (q < 0.05), whereas the dominant fatty acids (C16:0, C18:1n9c, C18:0, C18:2n6c) showed no proportional differences. KIRREL3 was most highly expressed in the kidney among eight tissues examined. These findings identify the 63-bp site as a candidate marker for meat quality in local Chinese cattle, with potential for marker-assisted selection pending confirmation in larger cohorts and functional studies.

Follicular fluid extracellular vesicles are beneficial for in vitro oocyte maturation and development; however, their effect on the expression profiles of oocyte microRNAs (miRNAs) and the roles of related miRNAs are unknown. In this study, we aimed to investigate miRNA expression in mature oocytes cultured in follicular fluid extracellular vesicles and the effect of miRNA-125a (miR-125a) on oocyte maturation. The expression profiles of the miRNAs were determined by microRNA sequencing, followed by target gene prediction analysis. We transfected miR-125a mimics and an miR-125a inhibitor to evaluate the effect of modulated miRNA-125a on cumulus expansion, oocyte maturation rate, changes in cytoplasmic maturation-related indicators, and changes in the expression of oocyte maturation-related, cumulus expansion-related, and predicted target genes. We found that miR-125a overexpression decreased the levels of cumulus expansion-related, oocyte maturation-related, and predicted target genes, adenosine deaminase RNA specific (ADAR), and lipid droplet number, and it increased the percentage of oocytes with abnormal cortical granule distribution. Inhibiting miR-125a increased the expression levels of oocyte maturation-related and target genes, number of lipid droplets, and endoplasmic reticulum function, and it decreased lipid droplet size. Mitochondrial membrane potential and reactive oxygen species levels were not significantly different between groups. In conclusion, our results suggest that extracellular vesicles may improve oocyte quality by modulating ADAR through regulating miR-125a.

Histone deacetylase 9 (HDAC9) is a histone deacetylase (HDAC) subtype IIa protein that deacetylates histone 3 (H3), histone 4 (H4), and nonhistone proteins in vivo to alter chromosomal shape and regulate gene transcription. There have been few studies on the regulatory influence of the HDAC9 gene on the differentiation of chicken embryonic stem cells (cESCs) into male germ cells, and the significance of HDAC9 is still unknown. Therefore, we explored the specific role of HDAC9 during differentiation of the cESCs of Jilin Luhua chickens through inhibition or overexpression. In medium supplemented with 10-5 mol/L retinoic acid (RA), cESCs were stimulated to develop into germ cells. HDAC9 and germline marker gene mRNA and protein levels were measured using qRT‒PCR and western blotting. During the differentiation of cESCs into male germ cells, overexpression of the HDAC9 gene greatly increased the mRNA and protein expression levels of the germline marker genes Stra8, Dazl, c-kit, and integrin ɑ6. The HDAC9 inhibitor TMP195 significantly decreased the mRNA and protein expression levels of the above markers. In summary, HDAC9 positively regulates the differentiation of cESCs.

In this study, we aimed to investigate the effect of the expression of miR-155-5p and its target genes on oocyte maturation. We analyzed the expression of miR-155-5p and its target genes in cumulus cells and oocytes using quantitative real-time reverse-transcription polymerase chain reaction. Using carboxyfluorescein, porcine cumulus cells were transfected with mimics and inhibitors of ssc-miR-155-5p to induce in vitro maturation, and subsequently, cumulus expansion, oocyte maturation, and cleavage rate were measured. We found that miR-155-5p expression in cumulus cells at the metaphase II stage was significantly higher than that at the germinal vesicle (GV) stage, whereas Cx43 expression was significantly lower than that at the GV stage (P < 0.05). Compared with those in the negative control group, the cumulus diffusion area of cumulus oocyte complexes; oocyte maturation rate; cleavage rate; HAS2, PTGS2, CD44, PTX3, and TNFAIP6 expression in cumulus cells; and GDF9, BMP15, CyclinB1, and CDK1 expression in oocytes were significantly increased in the miR-155-5p mimics group (P < 0.05), whereas the mRNA and protein expression of CX43 were significantly decreased (P < 0.05). Compared with that in the negative control group, the protein expression of CyclinB1 and p-CDK1 (Thr14, Tyr15) in the miR-155-5p mimics group was significantly increased (P < 0.05). These results suggest that miR-155-5p regulates maturation promoting factor activity by targeting Cx43, which improves the in vitro maturation and cleavage rate of porcine oocytes.

Context Muscle fibre characteristics are important internal factors that can directly affect pork quality. Especially muscle-fibre types can interconvert with the influence of certain factors.Aims The purpose of this experiment was to study the differences in meat quality among Songliao black pigs, Jilin Hua pigs and Dongliao black pigs, and the molecular mechanism of the differences.Methods First, the conventional meat-quality traits of each breed were determined. Second, the muscle-fibre characteristics of different muscles were analysed by the haematoxylin-eosin staining and immunohistochemistry. Next, the activity of several key oxidative/glycolytic enzymes was detected with kits. Finally, quantitative polymerase chain reaction was used to analyse the expression abundance of myosin heavy-chain (MyHC) genes and key genes related to muscle fibre-type transformation.Key results The results of meat-quality measurement showed that Songliao black pigs were superior to Jilin Hua pigs and Dongliao black pigs in water-holding capacity, tenderness and intra-muscular fat; and Jilin Hua pigs were superior to Dongliao black pigs in water-holding capacity. The diameter and area of muscle fibres were the smallest, the density was the largest, the proportion of oxidative muscle fibres, oxidative enzyme activity and expression abundance of oxidative genes were the highest in Songliao black pigs, followed by Jilin Hua pigs and Dongliao black pigs. The proportions of oxidative muscle fibres ranged from 10.37% to 33.6% in Songliao black pigs, from 6.96% to 26.42% in Jilin Hua pigs, and from 5.86% to 17.42% in Dongliao black pigs. The psoas major exhibited the smallest muscle-fibre diameter, the highest density, followed by triceps brachii, biceps femoris and longissimus thoracis. The oxidative muscle fibre proportions of the psoas major and triceps brachii were significantly greater than those of the biceps femoris and longissimus thoracis. The expression abundances of AMP-activated protein kinase (AMPK), silencing information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1 alpha) (Sirt1/AMPK/PGC-1 alpha) pathway and mitochondrial function-related genes were the highest in Songliao black pigs, followed by Jilin Hua pigs and Dongliao black pigs.Conclusions The results showed that Songliao black pigs had the best meat quality, followed by Jilin Hua pigs and Dongliao black pigs. The meat quality of psoas major and triceps brachii was significantly greater than that of biceps femoris and longissimus thoracis. This experiment suggests that transformation of oxidative muscle fibres can be promoted through the Sirt1/AMPK/PGC-1 alpha pathway.Implications By analysing the meat-quality traits and the molecular mechanism of meat-quality differences, it can provide data reference and direction for further meat-quality improvement of three breed pigs. We systematically studied the differences in meat quality among Songliao black pigs, Jilin Hua pigs and Dongliao black pigs, from the perspective of meat quality, muscle-fibre characteristics and Sirt1/AMPK/PGC-1 alpha signalling pathways. In addition, muscle-fibre characteristics of different parts (longissimus dorsi, psoas major, triceps brachii and biceps femur) of each breed were comprehensively studied instead of a single tissue. The results of oxidised muscle fibres were verified by immunohistochemistry, enzyme-activity assay and gene-expression methods.

为探索组蛋白去乙酰化酶9（Histone Deacetylase 9，HDAC9）基因的功能，明确其在芦花鸡不同组织中的表达差异，实验采集芦花鸡各组织，克隆获得HDAC9基因完整CDS区，进行生物信息学分析及组织表达分析。结果显示，芦花鸡HDAC9基因CDS区长度为3 210 bp，编码1 069个氨基酸；芦花鸡HDAC9基因氨基酸序列与號鹦鹉、绿头鸭及雉鸡位于一个分支，同源性较高，亲缘关系较近。HDAC9蛋白分子质量约为118.0 ku，分子式为C 5145 H 8261 N 1499 O 1613 S 33 ，理论等电点为6.33，半衰期为30 h，HDAC9蛋白为水溶性蛋白；HDAC9蛋白存在信号肽，为分泌蛋白；共存在67个潜在的磷酸化位点；存在3个N-糖基化潜在位点。HDAC9蛋白二级结构由α-螺旋、β-折叠、β-转角和无规则卷曲组成，三级结构预测结果与其一致。实时荧光定量PCR结果显示，HDAC9基因在芦花鸡不同组织中表达水平存在显著差异，其中在胸肌中表达量最高，其次为睾丸。本实验结果为进一步探索HDAC9基因生物学功能奠定基础。

1了解猪冷冻精液 1.1猪冷冻精液应用的优点 猪精液冷冻保存的最大优势是可以延长猪精液的使用期限.常温下新鲜精液使用时间有限,只能在1d内使用.在17℃下用常温稀释液处理的精液,猪精子的存活时间可达2周以上.利用液氮等为冷源,保存用冷冻稀释液处理的精液,只要精液处于液氮内,理论上可以无限期保存.

【Objective】 The aim of this study was to analyze the genetic diversity of forkhead box A2（FOXA2） gene exon 3 polymorphism in local pig breeds and its correlation with reproductive traits in Songliao Black pigs.【Method】 The single nucleotide polymorphisms（SNPs）（genotype frequency, gene frequency, genetic homozygosity, genetic heterozygosity, number of effective alleles, polymorphic information content） of FOXA2 gene exon 3 were screened by direct sequencing in ear tissues of 90 healthy Songliao Black pigs, and the correlation between SNP of FOXA2 gene eoxn 3 and reproductive performances（total number of litter, number born alive, number of nipples, birth weight, 3-week weight, weaning weight and weaned piglets） were analyzed using SPSS 26.0 software.【Result】 The results obtained that SNPs sites（C1337A,C1343G,C1626T）were found of FOXA2 gene exon 3 in Songliao Black pig.Genotypes of CC,CA and AA were contained in C1337A,genotypes of CC,CG and GG were contained in C1343G,and genotypes of CC,CT and TT were contained in C1626T.The chi-square fitness test showed that SNPs（C1337A,C1626T） were in Hardy-Weinberg equilibrium（P＞0.05）,C1343G was not in Hardy-Weinberg equilibrium（P＜0.05）.The calculation of polymorphism information content（PIC） showed that the genetic heterozygosities of C1337A and C1626T were in moderate polymorphism（0.25＜PIC＜0.50）,the genetic heterozygosity of C1343G was in the highly polymorphism（PIC＞0.50）.The results of association analysis showed that the total number born of CC genotype with C1337A locus was significantly higher than that of CA genotype（P＜0.05）,the 3-week weight and weaning weight of piglets born by AA genotype individuals were extremely significantly higher than that of CC and CA genotype（P＜0.01）;The 3-week weight and weaning weight of piglets born by CC genotype individuals with C1626T locus were significantly or extremely significantly higher than that of CT and TT genotype（P＜0.05 or P＜0.01）,the number of nipples of CC genotype was significantly higher than that of CT genotype（P＜0.05）.【Conclusion】 There were three SNPs sites（C1337A,C1343G,C1626T） in exon 3 of FOXA2 gene, which had a significant correlation with the total number born, 3-week weight, weaning weight and number of nipples of Songliao Black pig.

为了比较不同品种猪氨基酸组成的差异,试验采集松辽黑猪、新吉林黑猪、民猪背最长肌测定氨基酸组成和含量,并采用主成分分析和聚类分析比较三个品种猪氨基酸组成的差异.结果表明:新吉林黑猪必需氨基酸含量显著高于民猪(P＜0.05),松辽黑猪必需氨基酸含量与新吉林黑猪和民猪均差异不显著(P＞0.05);新吉林黑猪非必需氨基酸含量极显著高于民猪(P＜o.01),与松辽黑猪差异不显著(P＞0.05),松辽黑猪非必需氨基酸含量显著高于民猪(P＜0.05);松辽黑猪、新吉林黑猪呈味氨基酸、氨基酸总量均显著高于民猪(P＜0.05),新吉林黑猪与松辽黑猪间差异不显著(P＞0.05);三个品种猪必需氨基酸/氨基酸总量差异不显著(P＞0.05);民猪呈味氨基酸/氨基酸总量极显著高于新吉林黑猪(P＜0.01),显著高于松辽黑猪(P＜0.05).三个品种猪各种氨基酸之间存在显著(P＜0.05)与极显著正相关(P＜0.01).新吉林黑猪综合得分最高(2.68分),然后依次是松辽黑猪(0.84分)和民猪(-3.51分);松辽黑猪与新吉林黑猪聚为一类,民猪单独为一类.说明猪只肌肉氨基酸组成和含量受品种影响,新吉林黑猪氨基酸营养与风味好于松辽黑猪与民猪.

松辽黑猪2009年通过了国家畜禽遗传资源委员会的审定,获得猪新品种证书.该猪是吉林省农业科学院三代科学家经过23年,从1985年开始培育而成的具有自主知识产权的猪新品种.

The differentiation of embryonic stem cells (ESCs) into germ cells in vitro could have very promising applications for infertility treatment and could provide an excellent model for uncovering the molecular mechanisms of germline generation. This study aimed to investigate the differentially expressed miRNAs (DEMs) during the differentiation of chicken ESCs (cESCs) into male germ cells and to establish a profile of the DEMs. Cells before and after induction were subjected to miRNA sequencing (miRNA-seq). A total of 113 DEMs were obtained, including 61 upregulated and 52 downregulated DEMs. GO and KEGG enrichment analyses showed that the target genes were enriched mainly in the MAPK signaling pathway, HTLV infection signaling pathway, cell adhesion molecule (CAM)-related pathways, viral myocarditis, Wnt signaling pathway, ABC transporters, TGF-beta signaling pathways, Notch signaling pathways and insulin signaling pathway. The target genes of the miRNAs were related to cell binding, cell parts and biological regulatory processes. Six DEMs, let-7k-5p, miR-132c-5p, miR-193a-5p, miR-202-5p, miR-383-5p and miR-6553-3p, were assessed by qRT-PCR, and the results were consistent with the results of miRNA-seq. Based on qRT-PCR and western blot verification, miR-383-5p and its putative target gene STRN3 were selected to construct an STRN3 3'-UTR dual-luciferase gene reporter vector and its mutant vector. The double luciferase reporter activity of the cotransfected STRN3-WT + miR-383-5p mimics group was significantly lower (by approximately 46%) than that of the other five groups (p < 0.01). There was no significant difference in luciferase activity among the other 5 groups. This study establishes a DEM profile during the process of cESC differentiation into male germ cells; illustrates the mechanisms by which miRNAs regulate target genes; provides a theoretical basis for further research on the mechanisms of the formation and regulation of male germ cells; and provides an important strategy for gene editing, animal genetic resource protection and transgenic animal production.

为了研究中国草原红牛F-框蛋白32(FBXO32)基因多态性与肉质性状之间的相关性,试验选取120头中国草原红牛为研究对象,采用PCR产物直接测序法检测FBXO32基因内含子4的多态性,利用SPSS 19.0软件对多态性位点与离心失水率、滴水损失率、压榨水分、肌肉嫩度、肌内脂肪含量、初水分含量、蛋白质含量、大理石花纹评分、熟肉率、眼肌面积、pH24值指标进行相关性分析.结果表明:在FBXO32基因第4内含子g.16336603处检测到一个C/T突变位点,TT型为优势基因型,等位基因T为优势基因,该位点在所测群体中处于哈迪-温伯格平衡状态(P＞0.05);CC基因型离心失水率显著低于TT和TC基因型(P＜0.05),肌内脂肪含量和大理石花纹评分显著高于TT和TC基因型(P＜0.05),pH24值高于TT和TC基因型,其中与TC基因型差异显著(P＜0.05);但3个基因型的滴水损失率、压榨水分、肌肉嫩度、初水分含量、蛋白质含量、眼肌面积及熟肉率均差异不显著(P＞0.05).说明FBXO32基因可作为中国草原红牛品种选育标记.

为探究中国草原红牛介体复合物亚基4(MED4)基因多态性及其对草原红牛肉质性状的影响,实验以118头20月龄的草原红牛为研究对象,采用PCR测序的方法对草原红牛MED4基因第7外显子的多态性进行研究,利用SPASS 20.0软件分析MED4基因SNP位点与肉质性状的相关性.结果表明:草原红牛MED4基因第7外显子存在A268G、A420G突变,经卡方检验发现2处突变位点均处于Hardy-Weinberg不平衡状态,2处突变位点的遗传杂合度(He)相对较低,多态信息含量(PIC)介于0.25和0.5之间,属中度多态;对不同基因型对应个体的生产性状进行关联分析,发现A268G位点与肉质性状无显著相关,而A420G位点不同基因型个体间肌肉嫩度、pH和蛋白质含量存在显著差异.本实验可为MED4基因标记在草原红牛生产中的利用提供初步参考依据.

为了研究大白猪芳香烃受体(AHR)多态性与生长性状的关联,采用高分辨率熔解曲线(HRM)分型技术进行了基因分型,结合生长性状进行了相关性分析.提取246头大白猪耳组织DNA,进行混池测序,筛查多态性位点,针对发现的多态性位点重新设计引物,利用HRM分型法对个体进行多态性检测,结合已测定的生长性能数据进行关联分析.结果显示:检测到一个多态性位点38 332 C＞T,群体内存在C和T两个等位基因,CC、CT、TT三种基因型;x2检验表明该位点的基因型频率和等位基因频率在种群中分布差异显著,不符合Hardy-Weinberg平衡定律(P＞0.05);群体遗传学分析表明该位点在所检测群体中处于中度多态(0.25≤PIC≤0.5);关联分析发现该位点对6月龄体重和日增重的影响较为显著(P＜0.05).综上,AHR基因上的该位点多态性对大白猪的生长发育具有一定程度的影响,可为后续选育提供参考.

[目的]研究中国草原红牛丙酮酸脱氢酶激酶4(pyruvate dehydrogenase kinase 4,PDK4)基因多态性与肉质性状的关系.[方法]挑选120头中国草原红牛为研究对象,采用Sanger测序法检测PDK4基因第1～11外显子的单核苷酸多态性(SNP)位点,分析SNP位点的基因型频率、基因频率及群体遗传参数等.利用SPSS 21.0软件对中国草原红牛PDK4基因SNP位点多态性与肉质性状(熟肉率、肉嫩度、失水率、pH、滴水损失、肌内脂肪含量、初水分含量、蛋白质含量)进行关联分析.[结果]在中国草原红牛PDK4基因外显子8和外显子11上共检测到3个SNPs;PDK4基因第8外显子57 bp处存在1个SNP位点(G57C),且引起编码氨基酸的改变,存在GG、GC和CC 3种基因型;PDK4基因第11外显子在330和389 bp处存在2个SNPs位点(G330T和C389T),在G330T位点上存在GG、GT和TT 3种基因型;在C389T位点上存在CC、CT和TT 3种基因型.卡方适合性检验结果显示,中国草原红牛第8外显子G57C位点偏离Hardy-Weinberg平衡状态(P＜0.05),第11外显子的G330T和C398T符合Hardy-Weinberg平衡状态(P＞0.05);群体遗传参数分析发现,第8外显子G57C突变位点属于低度多态性位点(PIC＜0.25),等位基因数为1.1429,表明其在中国草原红牛中的变异较小;第11外显子G330T和C398T属于中度多态性位点(0.25＜PIC＜0.5),等位基因数分别为1.6431和1.6447,说明该遗传标记能够提供遗传信息.关联分析结果表明,PDK4基因第8外显子中G57C位点GG和CC基因型个体肌内脂肪含量显著高于GC基因型(P＜0.05);第11外显子G330T位点GG基因型滴水损失和初水分量显著高于GT基因型(P＜0.05);GG基因型肉嫩度显著高于TT基因型(P＜0.05);GT基因型肌内脂肪含量显著高于TT基因型(P＜0.05),C389T处CT基因型失水率显著低于TT基因型(P＜0.05).[结论]PDK4基因多态性与中国草原红牛肉质性状相关,可作为肉质性状的候选基因.

[目的]探究葛根素是否通过AMPK通路促进松辽黑猪前体脂肪细胞分化,以期为在饲粮中添加葛根素改善猪肉肉质提供参考.[方法]待松辽黑猪前体脂肪细胞汇合度达90％时进行为期4 d的诱导分化.诱导分化时,将细胞分为对照组(Con)、葛根素组(Pue)、葛根素+AMPK激活剂AICAR组(AICAR)及葛根素+AMPK抑制剂CompoundC组(CompoundC),对照组诱导液中不添加葛根素,Pue组添加40 μmol/L葛根素,AICAR组添加40 μmol/L葛根素+500 μmol/L AICAR,CompoundC 组添加 40 μmol/L 葛根素+20 μmol/L CompoundC.诱导分化结束,收集各组细胞,用甘油三酯(TG)酶法检测细胞中甘油三酯浓度;实时荧光定量PCR检测AMPK各亚基以及成脂分化相关基因的表达水平;用Western blotting检测过氧化物酶体增殖物激活受体γ(PPARγ)、AM P依赖的蛋白激酶(AMPK)、p-AMPK(Thr-172)、乙酰辅酶A羧化酶(ACC)的蛋白表达水平;通过Autodock Vina 1.20对葛根素、AMPKα亚基做分子对接预测.[结果]甘油三酯测定结果显示,与Con组相比,Pue组甘油三酯浓度显著增加(P＜0.05);与Pue组相比,AICAR组甘油三酯浓度显著降低(P＜0.05).实时荧光定量PCR结果显示,与Con 组相比,Pue 组 PPKAG2、PPKAB1、PPKAB2、PPKAA1、PGC-1α 表达量均显著降低(P＜0.05),Pue 组C/EBPα、PPARγ、ACC 表达量均显著增加(P＜0.05);与 Pue 组相比,AICAR 组 PPKAG1、PPKAG2、PPKAB1、PPKAB2、PPKAA1表达量显著增加(P＜0.05),AICAR组C/EBPα、PPARγ、ACC表达量均显著降低(P＜0.05),CompoundC 组 PPKAG1、PPKAG2、PPKAB2 表达量均显著降低(P＜0.05),CompoundC 组 C/EBPα、ACC 表达量均显著增加(P＜0.05).Western blotting结果显示,与Con组相比,Pue组PPARγ、ACC蛋白表达量显著增加(P＜0.05),AMPK、p-AMPK蛋白表达量显著下降(P＜0.05),且p-AMPK/AMPK显著下降(P＜0.05);与Pue组相比,AICAR组PPARγ、ACC蛋白表达量显著下降(P＜0.05),AMPK、p-AMPK蛋白表达量显著增加(P＜0.05),CompoundC 组 PPARγ、ACC 蛋白表达量显著增加(P＜0.05),AMPK、p-AMPK 蛋白表达量及 p-AMPK/AMPK显著下降(P＜0.05).[结论]40 μmol/L葛根素能够显著增加脂肪细胞中甘油三酯含量,显著上调成脂分化基因PPARγ、C/EBPα、ACC的表达量,显著下调AMPK各亚基的表达量,AMPK激活剂AIRCR可显著抑制葛根素的作用,说明葛根素可通过抑制AMPK通路调节松辽黑猪前体脂肪细胞成脂分化.

Mutton has recently been identified to be a consumer favorite, and intermuscular fat is the key factor in determining meat tenderness. Long-chain acyl-CoA synthetase 1 (ACSL1) is a vital subtype of the ACSL family that is involved in the synthesis of lipids from acyl-CoA and the oxidation of fatty acids. The amplification of the ACSL1 gene using rapid amplification of cDNA ends revealed that the alternative polyadenylation (APA) results in two transcripts of the ACSL1 gene. Exon 18 had premature termination, resulting in a shorter CDS region. In this study, the existence of two transcripts of varying lengths translated normally and designated ACSL1-a and ACSL1-b was confirmed. Overexpression of ACSL1-a can promote the synthesis of an intracellular diglyceride, while ACSL1-b can promote triglyceride synthesis. The transfection of ACSL1 shRNA knocks down both the transcripts, the triglyceride content was significantly reduced after differentiation and induction; and lipidome sequencing results exhibited a significant decrease in 14–22 carbon triglyceride metabolites. The results of the present study indicated that the ACSL1 gene played a crucial role in the synthesis of triglycerides. Furthermore, the two transcripts involved in various interactions in the triglyceride synthesis process may be the topic of interest for future research and provide a more theoretical basis for sheep breeding.

为研究中国草原红牛丙酮酸脱氢酶激酶4(Pyruvate Dehydrogenase Kinase 4,PDK4)基因功能,本研究克隆了PDK4基因编码区序列(CDS),从mRNA水平检测PDK4基因在中国草原红牛各组织及其前体脂肪细胞成脂分化过程中的表达差异,最后进行生物信息学分析.根据NCBI公布的牛PDK4基因序列(登录号:NM_001101883.1)设计引物,PCR扩增和克隆测序,获得草原红牛PDK4基因完整CDS序列,应用生物信息学软件分析蛋白理化性质和结构,预测蛋白信号肽和磷酸化位点及构建系统进化树;通过实时荧光定量PCR检测PDK4基因在中国草原红牛各组织中mRNA的相对表达量及PDK4基因在前体脂肪细胞分化过程中mRNA水平上的表达情况.结果显示:本研究成功获得中国草原红牛PDK4基因CDS序列,长度为1223 bp,编码407个氨基酸,与野牦牛同源性较高(99.8％),与埃及果蝠的同源性较低(89.7％).PDK4基因在草原红牛心脏、肝脏、脾脏、肺脏、肾脏、胃、十二指肠、背最长肌和脂肪中均显著表达,其中在背最长肌中的表达量显著高于其他组织.成脂分化结果表明,PDK4基因mRNA表达量在0～4 d呈上升趋势,4～6 d之间无显著差异.

Dermal papilla cells (DPCs) are the key dermal component of the hair follicle that directly regulates hair follicle development, growth and regeneration. Successfully isolated and cultured DPCs from Small-tail Han sheep could provide a good model for the study of hair follicle development mechanism in vitro. DPCs were isolated using enzyme digestion and dissecting microscope from Small-tail Han sheep. Adherent cells were identified by cell characteristics, particular gene expression, differentiation capability to adipocyte and osteoblast using specific differentiation mediums. Additionally, flow cytometry was used to detect the cell cycle of DPCs. Cells originating from the dermal papilla showed the morphological appearance of mesenchymal cells (fibroblast-like cells). Purified DPCs were positive for α-SMA (α smooth muscle actin) and vimentin; in addition to their strong proliferation abilities in vitro, these DPCs can be differentiated into adipocyte and osteoblasts lineage under appropriate culture condition. DPCs were successfully isolated and subcultured from Small-tail Han sheep, which exhibited progenitor cell features and multiple differentiation potency. It provides a material for studying the molecular mechanism of hair follicle development and hair cycle, which will promote wool production in the future.

A continuing effort in the development of non-peptide delta-opioid agonists as possible safe and effective pain relief agents has been seen in the last years. Novel potent and selective delta-opioid agonists based on the pirrolomorphinan framework have been designed, synthesised and biologically characterised in our laboratories. In particular, compound 7 (SB 235863) proved the concept that a selective delta-opioid agonist may be a valuable pain relief agent free of the unwanted side-effects usually associated with narcotic analgesics such as morphine.