谷物膳食纤维对高胆固醇饮食小鼠脂质代谢及肠道菌群的影响

本研究旨在探讨谷物膳食纤维(DF)对高胆固醇饮食小鼠的脂质代谢和肠道菌群的影响。选取C57BL/6J小鼠30只,并根据体重随机分为4组,分别饲喂基础饲粮(对照,CO组,n=8)、基础饲粮+5 g/kg DF(CO+DF组,n=8)、高脂高胆固醇饲粮(HF组,n=7)及高脂高胆固醇饲粮+5 g/kg DF(HF+DF组,n=7)。试验期8周,测定各组小鼠生长性能、血清和肝脏生化指标、粪便总胆固醇(TC)、短链脂肪酸(SCFA)、相关肝脏胆固醇和胆汁酸(BA)蛋白表达及肠道菌群。结果表明:1)CO+DF组的最终体重显著低于CO组(P<0.05),与HF组相比,CO组和HF+DF组的4周和最终体重均显著降低(P<0.05),肝脏和脂肪的重量显著减少(P<0.05)。且与HF组相比,DF+HF组摄食效率显著降低(P<0.05)。2)CO+DF组血清TC、甘油三酯(TG)含量及天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)活性以及肝脏TC和TG含量显著低于CO组(P<0.05),CO组和HF+DF组显著低于HF组(P<0.05),粪便TC含量仅在HF+DF组显著低于HF组(P<0.05)。3)DF对肝脏胆固醇调节元件结合蛋白-2(SREBP-2)、3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)、肝脏X受体α(LXRα)、微管相关蛋白1轻链3-Ⅰ(LC3-Ⅰ)和微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)蛋白表达没有显著影响(P>0.05)。CO+DF组肝脏胆固醇7-羟化酶(Cyp7a1)的蛋白表达水平显着低于CO组(P<0.05),但HF组和HF+DF组之间没有显著差异(P>0.05)。4)HF+DF组的粪便总SCFA和丙酸盐含量显著高于HF组(P<0.05)。与相应的对照组相比,CO+DF组和HF+DF组的粪便丁酸盐含量显著增加(P<0.05)。5)在门水平上,DF组厚壁菌门(Firmicutes)相对丰度减少,HF+DF组的拟杆菌门(Bacteroidetes)相对丰度高于HF组。梭状芽孢杆菌属ⅪⅤa(ClostridiumⅪⅤ)相对丰度在CO+DF组和HF+DF组显著增加(P<0.05)。此外,与其他组相比,仅HF+DF组中阿克曼菌属(Akkermansia)相对丰度显著增加(P<0.05)。由此可见,每千克饲粮添加5 g DF可降低饲喂含脂45%饲粮小鼠的体重和肝脏脂肪含量,降低血清TC、TG含量及AST和ALT活性以及肝脏TC和TG含量,降低胆固醇合成相关蛋白Cyp7a1的表达水平,提高总SCFA含量及肠道有益菌群和优势菌群相对丰度。     

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。     

不同剂量环磷酰胺对昆明小鼠肝肾组织的损伤作用研究

为研究不同剂量的环磷酰胺对小鼠肝肾组织及功能的损害程度,试验选取40只5周龄健康雌性昆明小鼠,随机分为5组,除对照组外,其余4组小鼠连续3 d分别腹腔注射40、80、120及160 mg/（kg·BW）环磷酰胺溶液,注射后第7天采集肝脏和肾脏组织样品及血清,制备石蜡组织切片及透射电镜切片,观察肝脏和肾脏的组织学变化,检测血清中甘胆酸（CG）、胆碱酯酶（ChE）、血清前白蛋白（PA）、总胆汁酸（TBA）、谷丙转氨酶（ALT）、谷草转氨酶（AST）、尿素氮（BUN）、血清肌酐（SCr）、尿酸（UA）含量/活性。结果显示,环磷酰胺可引起小鼠肝肾组织病理损害,呈剂量依赖性,肝脏比肾脏更早出现损伤。石蜡切片及透射电镜切片观察结果显示,环磷酰胺对肝脏的毒性作用主要表现在肝索紊乱、肝血窦扩张、微胆管淤胆、门管区及中央静脉周围炎性细胞浸润及纤维增生、组织间出血、肝细胞出现脂肪变性及气球样变性,凋亡及坏死细胞增多。肾脏组织损伤主要表现在出血及纤维增生,肾小管上皮细胞胞浆空泡样变、线粒体损伤、核固缩,上皮细胞基底膜增厚。环磷酰胺对膀胱的损害不严重。与对照组相比,40~160 mg/（kg·BW）剂量组小鼠血清中ChE活性及PA水平,120~160 mg/（kg·BW）剂量组AST活性,80~160 mg/（kg·BW）剂量组BUN水平及80 mg/（kg·BW）剂量组UA水平均显著升高（P<0.05）;其余生化指标与对照组差异不显著（P>0.05）。以上结果表明,环磷酰胺注射剂量在80~120 mg/（kg·BW）时可引起昆明小鼠肝脏和肾脏组织及细胞明显的病理损伤。该研究结果可为选择合适的环磷酰胺注射剂量用以制备动物肝脏免疫抑制模型提供试验依据。     

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     

基于梨枣轻微损伤的可见/近红外光谱判别研究

利用可见/近红外光谱技术对梨枣轻微损伤的分类判别建模方法进行研究。分别采用PLS-LDA(线性)和LS-SVM(非线性)建立判别模型,分析比较不同预处理方式和建模波段对模型精度的影响。结果表明:经9点平滑预处理后的短波近红外(780~1100nm)PLS-LDA模型判别效果最佳,校正集和预测集的正确识别率分别达到97.8%和96.7%。     

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.     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.