与鸭C4结合蛋白互作的鸭疫里默菌外膜蛋白的筛选及鉴定

旨在筛选并鉴定与鸭C4结合蛋白（C4b-binding protein，C4BP）互作的鸭疫里默氏菌（Riemerella anatipestifer，R.anatipestifer）外膜蛋白。本研究将保存的R.anatipestifer复苏培养，提取外膜蛋白，以鸭C4BPα作为诱饵蛋白进行His pull-down及LC-MS/MS蛋白质谱鉴定，筛选与鸭C4BP可能发生互作的候选外膜蛋白；将各候选蛋白进行克隆、原核表达，免疫小鼠制备多克隆抗体，利用Far-western blot验证与鸭C4BP发生相互作用的R.anatipestifer外膜蛋白；针对候选蛋白及C4BPα各功能结构域进行克隆及原核表达，利用Far-western blot鉴定候选蛋白及与C4BP的相互作用位点；利用ELISA对补体因子C3b、C4b及C4BP在R.anatipestifer表面沉积情况进行测定，验证候选蛋白的功能。结果显示，经His pull-down及LC-MS/MS蛋白质谱分析，共筛选出3个与鸭C4BP发生相互作用的R.anatipestifer外膜蛋白，即ECE-1、SODs和Omp62；成功获得3个外膜蛋白多克隆抗体，ELISA检测3种多克隆抗体效价均超过1∶6 400，Western blot检测3种多克隆抗体可以与重组蛋白发生特异性反应；Far-western blot结果显示，仅ECE-1能够与C4BP发生相互作用，并且只有ECE-1全长能与鸭C4BP相互作用，而鸭C4BP与ECE-1的相互作用区域位于C4BPα的SCR 2和SCR 3；当健康鸭血清稀释度为3.125%时，ECE-1抗体能够显著促进补体因子C3b、C4b在R.anatipestifer表面的沉积作用（P<0.05），当健康鸭血清稀释度为6.25%时，ECE-1抗体能够显著抑制C4BP在R.anatipestifer表面的沉积作用（P<0.05）。本研究成功筛选并鉴定出1个与C4BP互作的R.anatipestifer外膜蛋白ECE-1，为进一步阐明R.anatipestifer免疫逃逸机制奠定了基础。     

马齿苋水提物对热应激小鼠空肠结构及吸收功能的影响

旨在探讨马齿苋水提物（aqueous extract of Portulaca oleracea L.）对热应激小鼠空肠结构及吸收功能的影响。将40只昆明种小鼠随机分为4组（n=10）：空白对照组（C）、热应激组（HS）、马齿苋水提物组（AEP）、维生素C组（Vc），HS、AEP和Vc组每天于（40±1）℃环境下处理0.5 h，连续热应激6 d后将小鼠转置于室温下给药治疗。给药7 d后眶窦采血，并采集小鼠空肠；测定小鼠血清中D-木糖、葡萄糖含量；HE染色观察空肠组织病理学变化；qRT-PCR法检测空肠黏膜ZO-1、SGLT1及GLUT2 mRNA的相对转录水平。结果显示，与C组比较，HS组小鼠空肠黏膜绒毛高度极显著降低（P<0.01）；血清中D-木糖的含量极显著降低（P<0.01）；空肠黏膜中ZO-1、SGLT1和GLUT2 mRNA的相对转录水平均极显著降低（P<0.01）；与HS组相比，AEP组小鼠空肠绒毛高度极显著提高（P<0.01）；血清中D-木糖及葡萄糖的含量均显著升高（P<0.01或P<0.05）；空肠黏膜中ZO-1、SGLT1和GLUT2 mRNA的相对转录水平均显著升高（P<0.05或P<0.01）。高温处理可导致小鼠空肠黏膜脱落及吸收功能下降，而在给予马齿苋水提物治疗后可通过提高空肠ZO-1 mRNA表达量修复空肠黏膜结构，通过促进空肠黏膜SGLT1和GLUT2 mRNA的转录量改善吸收功能。马齿苋水提物能在一定程度上修复热应激导致小鼠空肠黏膜结构损伤，改善热应激小鼠肠道的吸收能力。     

Effects of DHRS3 in C2C12 Myoblast Differentiation and Mouse Skeletal Muscle Injury

Myoblast differentiation is an essential process during skeletal muscle development. C2 C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro. Dehydrogenase/reductase(SDR family) member 3(DHRS3) is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol. Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation. However, the effect of DHRS3 on mouse muscle cell differentiation was unclear. The objective of current study was to determine if DHRS3 affected muscle cell differentiation, and if DHRS3 was involved in muscle regeneration. Protein expression was determined by western blot and immunofluorescence analysis. The activation and inhibition of DHRS3 increased and decreased C2 C12 myoblast differentiation respectively, which indicated that DHRS3 could affect C2 C12 myoblast differentiation. DHRS3 expression was significantly changed during muscle regeneration, with the regeneration of muscle injury, the expression of DHRS3 tended to increase first and then decrease. It suggested that DHRS3 might be involved in muscle regeneration. In summary, this study confirmed the involvement of DHRS3 in C2 C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development.     

光响应性阳离子表面活性剂PCDA-C6-NH3+的合成及其对液滴蒸发行为的调控

农药的有效利用与农药液滴在植物叶片上的蒸发过程息息相关。传统表面活性剂虽然可以改变液滴的蒸发模式或速率,但并不能根据外界环境变化对蒸发过程进行主动调节。为实现"自主调控"农药液滴蒸发的目的,本文设计合成了一种含丁二炔官能团的光响应性阳离子表面活性剂(PCDA-C6-NH3+),利用紫外光刺激改变其在液滴中的聚集状态,调控液滴的蒸发过程。该表面活性剂以10,12-二十五碳二炔酸(PCDA)作为骨架,通过酰胺化反应引入末端氨基并质子化后得到。采用紫外-可见光谱研究了PCDA-C6-NH3+的拓扑化学聚合反应,并利用静态表面张力和动态接触角分析了其紫外光照前后的表面活性和蒸发行为。该研究为实现液滴"自主调控"蒸发过程提供了一种新思路,对提高农药利用率具有重要意义。     

民勤4种主要灌木植物δ13C季节变化及与气象因子的关系

对民勤4种主要灌木植物叶片/同化枝δ¹³C测定，研究不同荒漠灌木植物叶片/同化枝δ¹³C差异及其季节变化，并探讨植物叶片/同化枝δ¹³C与气象因子之间的关系。结果表明:1）4种灌木植物叶片/同化枝δ¹³C值不同月份之间存在极显著差异（P＜0.01），除梭梭外，其他3种植物δ¹³C值生长初期（5月）>生长中后期（6-10月）。4 种灌木植物叶片/同化枝δ¹³C值种间差异在5-10月都达到极显著水平（P＜0.01），沙拐枣＞梭梭＞柠条锦鸡儿＞白刺。2）4种主要灌木植物叶片/同化枝δ¹³C值受到物种、月份、物种×月份的影响，均达到极显著水平(P＜0.01)，但受物种的影响最大。3）4种灌木植物叶片/同化枝δ¹³C值与气象因子的关系因其物种而异；影响梭梭、沙拐枣、白刺、柠条锦鸡儿叶片/同化枝δ¹³C值的主要气象因子分别是平均气温、相对湿度、蒸发量。     

Spraying exogenous synthetic cytokinin 6‐benzyladenine following the waterlogging improves grain growth of waterlogged maize in the field

6‐Benzyladenine (6‐BA, a synthetic cytokinin) could improve the ability to resist adversity. However, very little attention has been given to its role in alleviating waterlogging damages on grain growth. A field experiment was performed to investigate effects of exogenous 6‐BA after waterlogging for 6 days at the third leaf stage (V3‐WL + 6‐BA) or the sixth leaf stage (V6‐WL + 6‐BA) on grain filling, and endogenous hormone of summer maize hybrids DengHai605 (DH605) and ZhengDan958 (ZD958), which were planted popularly in China. Exogenous 6‐BA alleviated waterlogging damages on grain filling by improving grain weight and volume, which was beneficial to yield increase. Grain yield of DH605 under V3‐WL + 6‐BA and V6‐WL + 6‐BA increased by 16% and 12%, respectively, compared with that of V3‐WL (waterlogging at the third leaf stage) and V6‐WL (waterlogging at the sixth leaf stage), while the corresponding values for ZD958 increased by 20% and 15%, respectively. Moreover, exogenous 6‐BA alleviated waterlogging damages on grain endogenous hormone content by increasing levels of endogenous indole‐3‐acetic acid, zeatin riboside, and gibberellic acid, and decreasing level of abscisic acid in grain of waterlogged summer maize during grain‐filling periods. Clearly, exogenous 6‐BA improved grain‐filling characteristics, and endogenous hormone, resulting in a significant yield increase of waterlogged summer maize.     

Effect of dietary vitamin B6 supplementation on growth and intestinal microflora of juvenile golden pompano (Trachinotus ovatus)

In order to investigate the dietary vitamin B₆ (VB₆) requirement for juvenile golden pompano, an experiment of six different diets with six dietary VB₆ levels (0, 3.75, 7.47, 10.6, 13.7 and 18.5 mg/kg) was conducted. The results indicated that the content of dietary VB₆ significantly increased weight gain rate (WGR), specific growth rate (SGR) and feed conversion ratio (FCR; p < 0.05). Golden pompano fed diet containing 7.47 mg/kg VB₆ achieved the maximum ALT, AST, GPx, hepatic VB₆ concentrations as well as the minimum MDA, however, the ALT, AST, GPx and hepatic VB₆ concentrations decreased while MDA increased with a further increase in dieatry VB₆. The highest values of GR and POD occurred at the 10.6 and 13.7 mg/kg dietary VB₆ (p < 0.05) respectively. A diet supplemented with 7.47 mg/kg VB₆ increased intestinal Na⁺, K⁺‐ATPase, Chymotrypsin, γ‐glutamyl transpeptidase, creatine kinase and amylase activities (p < 0.05). The relative abundance of Tenericutes and Bacteroidetes decreased while Proteobacteria and Firmicutes improved with an increase in dietary VB₆ levels up to 7.47 mg/kg. Quadratic regression analysis on WGR, hepatic VB₆ concentrations, AST and ALT indicated that the optimum dietary VB₆ levels for juvenile golden pompano were 8.84–9.28 mg/kg.     

Effects of dietary vitamin C on growth,flesh quality and antioxidant capacity of juvenile golden pompano Trachinotus ovatus

To investigate the effects of dietary vitamin C on growth, flesh quality and antioxidant capacity of juvenile golden pompano Trachinotus ovatus, a 56‐day feeding trial with five graded levels of dietary VC (D1: 11.69, D2: 34.89, D3: 59.10, D4: 114.26 and D5: 227.93 mg VC per kg of diet) was performed on 375 fish (triplicate groups of 25 fish per diet, initial weight 13.57 ± 0.09 g). Results showed that fish of D3 group exhibited the maximum specific growth rate (SGR) and the highest liver enzymatic activities of catalase (CAT) and glutathione peroxidase (GSH‐PX), which were consistent with the expression levels of cat and gsh‐px. Besides, the D3 group also showed higher contents of protein and lipid, and lower cooking loss, drip loss and malondialdehyde content in muscle than D1 group. The docosahexaenoic acid proportion in muscle increased with increasing dietary VC levels. Furthermore, the lowest expression levels of carnitine palmitoyltransferase1 (cpt1) and peroxisome proliferator‐activated receptor α (pparα) were detected in livers of D3 group. The optimum dietary VC level was 49.73 mg/kg from the broken‐line analysis based on the SGR, in which better growth performance, antioxidative ability and flesh quality were observed in T. ovatus juveniles.