外源海藻糖对NaHCO3胁迫下甘草幼苗生长调节及总黄酮含量的影响

本研究以甘草无菌苗为试验材料,采用植物组织培养的方法,分析50 mmol·L^-1 NaHCO₃ 胁迫下外源海藻糖对甘草幼苗生长量、叶绿素含量、渗透调节物质含量、抗氧化保护酶活性和总黄酮含量的影响。结果表明： 50 mmol·L^-1 NaHCO₃ 胁迫显著降低了甘草幼苗的生长量、叶绿体色素含量、K⁺ 浓度、抗氧化保护酶（SOD、POD、CAT）活性和总黄酮含量,显著提高了丙二醛、脯氨酸和可溶性糖含量以及Na⁺浓度;施加15 mmol·L^-1 海藻糖可显著提高甘草幼苗的生长量,提高叶绿素含量、K⁺ 浓度和总黄酮含量,降低丙二酸含量、脯氨酸含量、可溶性糖含量和Na⁺ 浓度,并且提高抗氧化保护酶活性。因此,NaHCO₃胁迫下施加外源海藻糖对甘草幼苗生长具有良好的调节作用,可以增强甘草的抗碱能力,促进甘草幼苗生长。本研究为外源海藻糖提高甘草耐碱性和揭示其调控机制提供理论依据。     

绒山羊皮肤毛囊兴盛前期组织形态、chi-miR-107-3p与RHBDF2基因表达分析

为了探究chi-miR-107-3p和菱形家族蛋白2（rhomboid family member 2,RHBDF2）基因在不同品种（系）和光控增绒绒山羊兴盛前期（5~7月份）皮肤毛囊重建时的表达差异,本试验在7月份采集内蒙古阿拉善型绒山羊、敏盖绒山羊和光控增绒技术处理的阿尔巴斯型绒山羊体侧皮肤毛囊组织,采用组织切片技术对组织形态比较分析,实时荧光定量PCR法检测chi-miR-107-3p和RHBDF2基因的表达量。结果显示,在皮肤毛囊兴盛前期,阿拉善型绒山羊次级毛囊正处于重建阶段,敏盖绒山羊和阿尔巴斯型绒山羊（光控增绒）次级毛囊重建已基本完成;在阿拉善型绒山羊皮肤组织中chi-miR-107-3p表达量极显著高于敏盖绒山羊和阿尔巴斯型绒山羊（光控增绒）（P<0.01）,而RHBDF2基因表达量极显著低于其他两品种（系）（P<0.01）,且RHBDF2基因表达量在敏盖绒山羊皮肤组织中显著低于阿尔巴斯型绒山羊（P<0.05）。不同品种（系）和光控增绒绒山羊皮肤毛囊兴盛前期组织显微结构与chi-miR-107-3p、RHBDF2基因在皮肤组织中的表达差异分析结果相一致,次级毛囊重建初期chi-miR-107-3p表达量较高,而RHBDF2基因表达量极低,随着次级毛囊重建完成,chi-miR-107-3p表达量明显降低,RHBDF2基因表达量逐渐增高,且在不同品种（系）内蒙古绒山羊绒毛生长发育过程中的表达机制基本相同。因此,chi-miR-107-3p和RHBDF2基因是绒山羊皮肤毛囊生长发育的重要调控因子。     

猪戊型肝炎病毒ORF3蛋白影响HepG2细胞核黄素代谢的lncRNA-mRNA调控网络的鉴定

为初步鉴定并挖掘出猪戊型肝炎病毒(Hepatitis E virus,HEV)ORF3蛋白影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络,本试验通过构建腺病毒过表达载体,制备高滴度过表达腺病毒,介导猪 HEV-ORF3在HepG2细胞中实现过表达。Western blotting检测ORF3蛋白过表达成功后,运用lncRNA高通量组学测序,筛选出差异表达的lncRNA并进行靶向差异基因预测,对lncRNA靶向差异基因进行GO功能和KEGG通路富集分析,初步鉴定出与核黄素代谢信号通路相关的lncRNA-mRNA调控网络。Western blotting结果显示,在约为12 ku处出现目的条带,说明成功实现腺病毒介导猪HEV-ORF3在HepG2细胞中过表达。lncRNA高通量组学测序结果显示,共发现102个显著差异表达lncRNAs表达量上调,80个显著差异表达lncRNAs表达量下调。GO功能和KEGG通路富集分析显示,初步挖掘出lncRNA(MSTRG.13995.2)和lncRNA(MSTRG.1960.1)可能是与核黄素代谢信号通路相关的显著差异表达lncRNA,分别通过顺式调控其靶向基因APC-5和FLAD1来影响核黄素代谢信号通路。本试验初步鉴定出lncRNA(MSTRG.13995.2)-APC5和lncRNA(MSTRG.1960.1)-FLAD1可能是影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络。     

延边黄牛MYH3和MYH8基因多态性及其与生长性状的关联分析

本研究旨在探索延边黄牛肌球蛋白重链3（MYH3）和肌球蛋白重链8（MYH8）基因多态性及其与生长性状的关联性,应用DNA直接测序法和高分辨率熔解曲线分型技术对120头24月龄延边黄牛MYH3、MYH8基因的遗传变异进行分析,并结合延边黄牛生长性状数据进行了关联分析。结果显示,在MYH3基因的29602748位点检测到一处T>G突变,包含2个等位基因：T和G,存在3种基因型：TG、TT和GG;在MYH8基因的29406042位点检测到一处C>T突变,包含2个等位基因：C和T,存在3种基因型：CT、CC和TT。关联分析显示,MYH3基因29602748位点T>G突变显著影响24月龄延边黄牛十字部高和腰角宽,表现为GG基因型十字部高显著高于TT基因型（P<0.05）,腰角宽显著高于TT和TG基因型（P<0.05）。MYH8基因29406042位点C>T突变显著影响24月龄延边黄牛体重和胸围,表现为CT和TT基因型体重显著高于CC基因型（P<0.05）;TT基因型胸围显著高于CC基因型（P<0.05）。本研究结果表明,MYH3、MYH8基因多态性与延边黄牛的部分生长性状相关,可作为现代肉牛育种中分子标记辅助选择的候选基因。     

3D打印技术在水利水电工程施工中应用研究

对3D打印技术进行应用研究,阐述其精度、强度和速度等关键性能指标合理性,能够有效避免特殊形式结构部位的施工带来的成本、工期及质量问题,增加工程效益,并对溢流堰面特殊部位计算表面曲线轮廓,进行实验3D打印,最后总结3D打印在水利水电工程施工中的优势。     

Isolation and Identification of BPIV3 Genotype C

To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg²⁺ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.     

Expression Analysis of THBS3 Gene in Different Tissues and Skeletal Muscles at Different Periods from Landrace and Tongcheng Pigs

To study the expression pattern of THBS3 gene in different tissues and during skeletal muscle development, the THBS3 gene expression in different tissues and skeletal muscles during prenatal periods (33, 45, 65, 70 and 90 d) and postnatal periods (0, 9, 30, 60, 120 and 160 d) from Landrace and Tongcheng pigs were detected by Real-time quantification PCR.The results showed that THBS3 gene widely expressed in all tissues examined, exhibiting similar spatial expression patterns with expression peaks in lung in both pig breeds except in stomach and intestine.Moreover, although THBS3 gene showed a significant higher expression level in gestation than after birth in Landrace and Tongcheng pigs (P<0.05), it exhibited different expression patterns between Landrace and Tongcheng pigs, the expression peak was detected at gestation day 45 in Landrace pig, while was detected at gestation day 65 in Tongcheng pig.The results suggested that THBS3 gene involved in skeletal muscle growth and development in pigs, as well as the regulation of asynchronization of skeletal muscle development in different pig breeds.     

Basic characterization of avian β‐defensin genes in the Japanese quail,Coturnix japonica

In this study, we identified a cluster of 14 avian β‐defensins (AvBD; approximately 66 kbp) in the Japanese quail, Coturnix japonica. Except for AvBD12 (CjAvBD12) and ‐13, the CjAvBDs coding sequences exhibited greater than 78.0% similarity to the respective orthologous chicken AvBD genes (GgAvBD). The putative amino acid sequence encoded by each CjAvBD contained six cysteine residues and the GXC (X1‐2) motif considered essential for the β‐defensin family. Each CjAvBDs also formed a sub‐group with the respective orthologous genes of various bird species in a phylogenetic tree analysis. Synteny between the CjAvBD cluster and GgAvBD cluster was confirmed. The CjAvBD cluster was mapped on the long‐arm end of chromosome 3 by linkage analysis based on single nucleotide polymorphisms (SNPs) of CjAvBD1 and CjAvBD12 (approximately 46kbp), as well as GgAvBD cluster. We also confirmed that CjAvBD1, ‐4, ‐5, ‐9, and ‐10 are transcribed in 20 tissues, including immune and digestive tissues. However, our experimental data indicated that the CjAvBD cluster lacks the AvBD3 and ‐7 loci, whereas the CjAvBD101α, ‐101β, and ‐101θ loci arose from gene duplication of the AvBD6 orthologous locus in the CjAvBD cluster after differentiation between Coturnix ‐ Gallus.     

Increased humoral antibody response of foot-and-mouth disease virus vaccine in growing pigs pre-treated with poly-γ-glutamic acid

This study was conducted to determine if humoral antibody response of foot-and-mouth disease (FMD) vaccine improved in 8-week-old growing pigs born to well-vaccinated sows pre-treated with 60 mg of poly-γ-glutamic acid (γ-PGA) three days before vaccination. Antibody against FMD virus serotype O was measured 0, 2, 4 and 6 weeks post-vaccination, using a PrioCHECK FMDV type O ELISA kit. The results showed that positive antibody reactions against FMDV serotype O antigen among a component of the vaccine significantly increased in response to pre-injection with γ-PGA.