干扰TP53INP2抑制牛成肌细胞分化

【背景】肌肉可以维持哺乳动物运动功能并调节机体代谢,它的数量和分布对肉质有重要影响,骨骼肌的生长发育及其遗传特性在很大程度上影响甚至决定家畜的产肉量和肉品质,研究骨骼肌的生长发育具有重要意义。TP53INP2对自噬的调控作用以及对前体脂肪细胞分化的调控机制已有研究,而对于牛成肌细胞分化的影响尚未报道。【目的】探究TP53INP2对秦川牛成肌细胞分化的影响,为肉牛肉用性状分子育种工作提供理论依据。【方法】利用实时荧光定量PCR（real-time fluorescence quantitative PCR,RT-qPCR）技术检测了TP53INP2在24月龄秦川牛不同组织中的表达特性,同时分析了其在体外培养的牛骨骼肌成肌细胞不同分化时期的表达规律;合成TP53INP2基因siRNA并转染秦川牛成肌细胞,转染后12h进行诱导分化,观察成肌细胞表型变化,并利用RT-qPCR技术和Western Blot技术分别检测其在诱导分化第四天时分化标志基因和蛋白的表达情况。【结果】1.RT-qPCR结果显示,TP53INP2在成年秦川牛背最长肌组织中表达量最高,在小肠组织中表达量最低。TP53INP2在成年牛心脏、肝脏、肾脏、网胃、瘤胃中表达量较高,在其余组织中表达量较低（与背最长肌相比）。2.随着成肌细胞的分化,该基因在第0—4天表达量呈上升趋势且在第4天达到峰值,随后表达量呈下降趋势。3.在成肌细胞中干扰TP53INP2后,试验组肌管的数量和长度均显著低于对照组。4.干扰TP53INP2并提取细胞总RNA,RT-qPCR结果显示,成肌细胞分化标志基因肌细胞生成素（MYOG）和肌球重链蛋白3（MYH3）相对于对照组表达量显著降低。5.干扰TP53INP2并提取细胞总蛋白,Western Blot结果显示,试验组MYOG、MYH3、MYOD的蛋白表达量均降低且与对照组相比差异极显著。【结论】干扰TP53INP2对牛成肌细胞的分化具有抑制作用,提示该基因可能对秦川牛肌肉组织的生长发育具有重要的调控作用,可对其进行深入的功能研究以期用于肉牛的分子育种实践中。     

β-carotene attenuates weaning-induced apoptosis via inhibition of PERK-CHOP and IRE1-JNK/p38 MAPK signalling pathways in piglet jejunum

Weaning may cause oxidative injury, immune response impairment, apoptosis and other injuries in piglets. Oxidative and endoplasmic reticulum stress (ERS) can elicit inflammatory responses, and persistent oxidative and ERS also may lead to apoptotic cascades, which is associated with the pathogenesis of multiple diseases. β-carotene, a natural carotenoid, has potential anti-inflammatory and antioxidant functions. However, the effect of β-carotene on apoptosis in weaned piglets and the detailed molecular mechanism remain unclear. In this study, we found that β-carotene decreased malondialdehyde (MDA) levels and increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in piglet serum. β-carotene could inhibit the mRNA levels of caspase-3 significantly, but had no significant inhibitory effect of the mRNA levels of caspase-9 and caspase-12 in the piglet jejunum. In addition, β-carotene decreased the activation of GRP78, CHOP, and JNK/p38 MAPK and the ratio of Bax/Bcl-2. Furthermore, β-carotene had a significant influence on the activation of ERS and apoptosis-related signals in TG-induced IPEC-J2. In the present study, β-carotene pre-treatment attenuated the ratio of Bax/Bcl-2 and prevented TG-induced increases in the level of PERK-CHOP and IRE1-JNK/p38 MAPK pathway activation in a dose-dependent manner. Overall, these findings indicate that β-carotene may protect weaning-induced apoptosis through inhibiting ERS.     

适用于转录组测序的大白菜小孢子总RNA提取方法筛选

为了找到适用于转录组测序要求的大白菜小孢子总RNA 提取方法，以出胚率较高的吉红82 为试材，以热激诱导处理后的小孢子为研究对象，比较分析了TRIzol 法、改良CTAB 法及超纯试剂盒法3 种方法的RNA 提取效果。结果表明：TRIzol法（磨样）和改良CTAB 法提取的RNA 浓度较高，但完整性较差；超纯RNA 提取试剂盒法提取的RNA 质量最高，且完整性好，能够满足转录组测序的要求。     

山核桃天然群体家系种实和幼苗性状变异分析

基于岛石天然林山核桃自由授粉产生的44个单株果实与种子的表型及由种子实生繁殖1年生幼苗生长性状11个指标的数据,分别计算了各指标的特征值,并进行了方差分析,研究其自然变异.结果表明,果实和种子各表型变异程度不一,其中以果实质量的变异系数最大,为11.22％;其次是种子质量和出籽率,分别为8.64％和7.66％;种实各指标在采样单株间的差异均极显著.除果实质量与果形指数和种形指数之间无显著相关外,其余各指标两两间相关均极显著.1年生苗木苗高和地径变异系数分别达13.99％、10.28％,且各家系间差异极显著;苗高与地径的Pearson相关系数为0.552,相关性极显著.2个苗木生长指标与9个种实表型指标间相关性不显著.     

上海市小白菜高效安全生产现状与建议

上海市设施土壤栽培小白菜生产管理现状调查表明,本地区小白菜生产存在种子成本高、水肥管理粗放、生产效率低下等问题,建议继续加强小白菜高效安全生产模式建设,推广种子加工处理、水肥一体化、病虫害绿色防控关键生产环节机械化等技术的应用。     

Ultrastructural impairment of islet microvascular endothelial cells in STZ-induced type 1 diabetic mice

AIM: To investigate the ultrastructural changes of islet microvascular endothelial cells in STZ-induced type 1 diabetic mice. METHODS: BALB/c mice were randomly divided into diabetic group and control group. The expression of insulin and platelet-endothelial cell adhesion molecule-1 (CD31) in islet microvessels was detected by immunohistochemical staining. The ultrastructural changes of islet β cells and islet microvessels were observed under transmission electron microscope. RESULTS: Compared with control group, the number of islet β cells, ratio of β cells/α cells, average number of secretory granules in β cells and insulin expression area per islet in diabetic group were significantly decreased (P<0.01). Besides, diabetic group had fewer microvessels with lower expression of CD31 (P<0.01). Mitochondria in islet microvascular endothelial cells and pericytes in diabetic group were swelling. The basement membrane of islet microvessels became thicker in diabetic group (P<0.01). CONCLUSION: Islet microvascular endothelial cells were impaired in type 1 diabetic mice.     

长期培肥我国典型黑土玉米氮肥效应的演变趋势

【目的】依托中国东北典型农田设置的公主岭黑土长期培肥试验,研究长期培肥条件下黑土地区玉米产量和氮肥效应的时间演变趋势,阐明氮肥利用率和土壤养分之间的相关性,为指导我国建立长期培肥模式和提高氮肥利用率提供依据。【方法】本文选择不同有机肥用量(M0、M30和M60)下不施肥处理(CK)、施氮肥(N)、施磷钾肥(PK)和平衡施肥(NPK),共计12个处理,研究长期培肥对玉米的氮肥农学效率、氮肥当季利用率和氮肥累积利用率的影响。【结果】有机肥料和化肥(NPK)对玉米均具有显著增产效果,有机肥化肥配施可降低玉米产量的变异系数(CV)和提高可持续性指数(SYI)从而保证玉米的持续稳产高产。平衡施肥条件下的氮肥农学效率、当季利用率和累积利用率随着施肥年限的延长呈上升趋势并且达到显著或极显著水平;常量有机肥区和高量有机肥区的土壤肥力得到显著提升,氮肥效应随时间演变趋势分别为变化不显著和显著降低趋势。氮肥利用率与土壤有机碳、土壤全氮、土壤全磷和土壤速效钾呈显著负相关。【结论】在相对较高的肥力条件下,黑土地区玉米氮肥效应呈显著下降或变化不显著的趋势将为高肥力条件下有机肥部分替代化肥提供科学依据。     

Preliminary Study on NF-κB Signaling Pathway Regulating Brucella Intracellular Survival Molecular Mechanisms

By the infection of Brucella virulent strain and attenuated strain in mice macrophage RAW264.7,the assay was aimed to explore the relationship between NF-κB signaling pathways and Brucella virulent strain and attenuated strain in intracellular survival.Use different MOI Brucella (2308,RB51,16M and M5) to infect mice macrophage RAW264.7,after 0,4,8 and 24 h infected,cracking cell and collecting supernatant,we detected the effect of Brucella on activation of NF-κB signaling pathway by Western blotting.Different concentrations of NF-κB signaling pathway inhibitor were incubated with mice macrophage RAW264.7,with different multiplicities of infection (MOI) of Brucella infecting cells,ELISA kits to detect the expressions of TNF-α,IL-1β and IL-6 cytokine;At the same time,count the number of intracellular bacteria of CFU.The results showed that rough cattle Brucella strains RB51 could strongly activate NF-κB signaling pathway,smooth cattle Brucella strains 2308 was weak in the activation;At the same time,the activation of NF-κB signaling pathway was concentration dependent.When the MOI was 80,infection time was 8 h,NF-κB activation degrees of rough cattle Brucella strains RB51 and smooth cattle Brucella strains 2308 were the strongest,and this pathway was involved in producing TNF-α and IL-6;NF-κB signaling pathway inhibitor BAY11-7082 affected Brucella intracellular survival.So rough cattle Brucella strains RB51 intracellular survival and NF-κB signaling pathway activity were closely related.The results laid the foundation for the further study of Brucella intracellular pathogenesis,also provided scientific basis for the research of new drugs to Brucella,and prevention and treatment of brucellosis.     

西藏查吾拉地区不同性别牦牛产肉性能和肉营养成分的比较

为了研究性别对查吾拉地区牦牛的产肉性能及肉营养成分的影响,随机选取8岁健康的查吾拉牦牛10头,公母各半,对不同组别间牦牛进行屠宰试验比较肉营养品质的差异。结果表明,母牦牛屠宰率、净肉率、胴体率、骨肉比均高于公牦牛,但无统计学差异。肉营养成分分析结果显示,母牦牛肉中维生素A、E、B₁₂、半胱氨酸、组氨酸、精氨酸、钙、硒、脂肪酸含量高于公牦牛(P<0.05)。研究结果表明,查吾拉牦牛富含较高的营养成分,可为人类提供优质健康的动物蛋白,具备作为优质牦牛肉源生产的潜力。相比较,母牦牛肉优于公牦牛。