厄贝沙坦及其复方制剂中痕量杂质N-亚硝基-N-甲基-4-氨基丁酸残留方法学研究

采用超高效液相色谱-三重四级杆质谱(UPLC-MS/MS)建立厄贝沙坦及其复方制剂中N-亚硝基-N-甲基-4-氨基丁酸(NMBA)的测定方法。采用Waters ACQUITY UPLC○RHSS T3色谱柱进行分离,以0.005 mol/L甲酸铵(用甲酸调pH值至3.0)为流动相A,甲醇为流动相B,梯度洗脱,质谱检测器,流速为0.3 mL/min。结果显示NMBA的线性范围为0.156 6~3.132 6 ng/mL,相关系数r>0.99,方法检出限浓度为0.047 0 ng/mL(相当于样品浓度0.01μg/g),定量限浓度为0.156 6 ng/mL(相当于样品浓度0.03μg/g),样品加标回收率在80%~130%。说明该方法可以快速、简便、灵敏、准确地测定厄贝沙坦及其复方制剂中NMBA含量。     

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究

试验旨在研究伪狂犬病病毒(PRV)在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞(3D4/21)p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。     

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究

试验旨在研究伪狂犬病病毒（PRV）在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞（3D4/21）p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。     

抗旱、耐盐碱、矮秆陆地棉数量性状的遗传相关及主成份分析

对从国内外引进的抗旱、耐盐碱、矮杆棉花资源材料 ,经抗旱、耐盐碱、矮杆性鉴定和比较 ,选取了其中表现较好的 1 9份材料 ,进行了遗传相关和主成份分析 ,结果表明 :在 5 0 mmol· L- 1Na Cland1 0 0 mmol· L- 1Na Cl胁迫下的出苗率间呈正相关 ,抗盐性与产量间呈负相关 ;第一棉铃高度与纤维品质性状呈正相关。抗旱、耐盐碱、矮杆棉花材料的前 6个因子贡献率达 86.5 % ,其分别为第一棉铃高度因子、果枝数因子、单株结铃数因子、矮杆因子、单铃重因子等     

灌溉设备产品信息与报价系统的开发

本文介绍了开发灌溉设备产品信息与报价系统的必要性及功能 ,较系统地阐述了该软件系统的实现思路 ,并对其应用前景作了初步预测     

Research Progress on Function of Pig Bactericidal/Permeability-increasing Protein (BPI) and Its Application in Resistance Breeding

Bactericidal/permeability-increasing protein (BPI) has a strong effect on sterilization (mainly for G^- bacteria),neutralizing the activity of lipopolysaccharide (LPS) and enhancing the phagocytosis of mononuclear cells and neutrophils to pathogenic bacteria.The biological functions of BPI have been researched widely in recent years,which is known as "super antibiotic" and has been explored by many scholars as a candidate gene for resistance.This author summarized the research progress and application prospect of the BPI gene in the pig resistant breeding by introducing the structure,biological function of BPI gene and its relationship with the resistance,which was aimed to provide theoretical references and basis for the function research of pig BPI gene and its practical application in resistance breeding in future.     

Effects of different dl‐selenomethionine and sodium selenite levels on growth performance,immune functions and serum thyroid hormones concentrations in broilers

This trial was conducted in a 2 × 3 + 1 factorial arrangement based on a completely randomized design to evaluate the effects of different dl ‐selenomethionine (dl ‐Se‐Met) and sodium selenite (SS) levels on growth performance, immune functions and serum thyroid hormones concentrations in broilers. A total of 840 Ross 308 broilers (7 days old) were allocated by body weight to seven treatments (three replicates of 40 birds each treatment) including (1) basal diet (containing 0.04 mg of selenium (Se)/kg; control) without supplementary Se; (2, 3 and 4) basal diet + 0.05, 0.15 or 0.25 mg/kg Se as SS; (5, 6 and 7) basal diet + 0.05, 0.15 or 0.25 mg/kg Se as dl ‐Se‐Met. The experiment lasted 42 days. The results revealed that dietary Se supplementation improved (p < 0.05) average daily gain, feed efficiency, immune organ index, serum immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM) and triiodothyronine (T₃) concentrations and decreased (p < 0.01) thyroxine (T₄)/T₃ ratio in serum compared with the control. Broilers receiving the dl ‐Se‐Met‐supplemented diets had higher (p < 0.05) feed efficiency, thymus index, the amounts of IgA, IgG, IgM and T₃ as well as lower (p < 0.05) serum T₄ concentrations and T₄/T₃ ratio than those consuming the SS‐supplemented diets. Serum IgA and IgM levels of broilers fed 0.15 mg Se/kg were significantly higher (p < 0.05) than those of broilers fed 0.05 or 0.25 mg Se/kg. In summary, we concluded that dl ‐Se‐Met is more effective than SS in increasing immunity and promoting conversion of T₄ to T₃, thus providing an effective way to improve the growth performance of broilers. Besides, based on a consideration of all experiment indices, 0.15 mg Se/kg was suggested to be the optimal level of Se supplementation under the conditions of this study.