5个转Bt基因抗虫水稻恢复系的品质配合力分析

使用5个转Bt基因水稻恢复系作为父本,以12个不育系作为母本,根据5×12(NCII)不完全双列杂交设计配制60个组合,分析了这些组合的9个稻米品质性状的配合力及遗传参数.结果表明:精米率主要受基因加性效应的影响,整精米率、垩白粒率、碱消值、胶稠度主要受基因非加性效应互作的影响;除精米率主要受母本影响外,其余8个性状均受双亲交互作用的影响;垩白粒率和垩白度在后代遗传中主要受基因遗传作用的影响,可以在早代直接选择.配合力方差分析结果表明:精米率、垩白度及碱消值受环境因素的影响较大;昌恢891T为一般配合力较好的父本,华1165S为一般配合力较好的母本;原香39A/昌恢891T、泰乡1209A/昌恢T025T、昌盛843A/昌恢T025T为较优的组合,其中原香39A/昌恢891T的特殊配合力的综合评价最好.     

河南油田注聚井聚合物堵塞机理及防治技术

为解决河南油田聚合物驱油注聚井堵塞导致采收率严重下降的问题,采用物理模型试验及扫描电镜(SEM)的方法对堵塞位置、堵塞因素进行研究.聚合物注入性试验结果表明,注入阻力越大,注入压力就越高,确定堵塞位置为近井地带,结果与实际矿场结果相符;扫描电镜结果表明,高渗岩心以吸附堵塞为主,低渗岩心以机械捕集为主;静态吸附试验显示防吸附剂质量分数、聚合物相对分子质量、矿化度、温度均对聚合物吸附有不同程度影响,为此研制出一种使砂岩表面润湿反转以减少吸附的防吸附剂(代号5807),并对注入参数进行优化:防吸附剂最优注入质量分数为0.5％;该防吸附剂耐盐性较好,不同矿化度(1500～10000mg/L)对该防吸附剂体系的防吸附率影响不大;防吸附剂注入体积数越大,聚合物防吸附率越高,最佳注入体积为0.6PV;防吸附剂处理地层的时间越长,聚合物防吸附率越高,处理时间在12h以上防吸附率大于70％;防吸附剂的最佳注入方式为:顶替液+0.6 PV防吸附剂段塞+1PV清洗剂.该研究对油田后续现场施工及动态调整具有重要指导意义,同时也可为河南油田提高聚合物驱技术应用效果提供理论支持.     

农村互联网发展对中国玉米全要素生产率的影响

采用DEA-mamlquist指数测算2004—2019年中国17个玉米主产省(区)的全要素生产率,运用联立方程组模型实证检验农村互联网发展对玉米全要素生产率的影响及其作用机理,并分区域探讨其差异性。结果显示:2004—2019年中国玉米全要素生产率年均增长0.2%,主要依靠技术进步的单轨模式驱动。农村互联网的发展显著(P<0.01)提升玉米全要素生产率,主要依靠技术进步和技术效率的协同作用驱动。分区域来看,农村互联网发展对玉米全要素生产率均具有显著(P<0.01)的促进作用,其影响程度由高到低依次为北方春播玉米区>黄淮海平原夏播玉米区>西北灌溉玉米区>西南山地玉米区。建议进一步提高农村互联网的配套设施建设,发挥互联网“连接经济”的优势,应用多元化互联网技术,促进不同生态类型区玉米生产效率的提升。     

饲养密度和硫辛酸对肉鸡血清指标、行为及骨骼发育的影响

本试验旨在研究高饲养密度条件下饲粮中添加硫辛酸(LA)对肉鸡血清指标、行为及骨骼发育的影响。试验选取健康状况良好、体重相近的1530只21日龄爱拔益加(AA)肉鸡,随机分成3组,即常规饲养密度组(ND组,15只/m^2,饲喂基础饲粮)、高饲养密度组(HD组,18只/m^2,饲喂基础饲粮)和高饲养密度+LA组(HD+LA组,18只/m^2,在基础饲粮在添加300 mg/kg LA),每组6个重复。试验期为3周。结果表明:1)与ND组相比,HD组肉鸡血清钙含量和碱性磷酸酶(ALP)活性显著下降(P<0.05),血清皮质酮(CORT)含量以及还原型辅酶Ⅱ氧化酶(NOX)和髓过氧化物酶(MPO)活性显著上升(P<0.05)。2)与ND组相比,HD组肉鸡行走、梳翅、采食频次与时间占比显著降低(P<0.05),而饮水频次以及饮水和趴卧时间占比显著升高(P<0.05)。3)与ND组相比,高饲养密度显著降低肉鸡胫骨与股骨直径、重量、密度、强度以及钙、磷、灰分和矿物盐含量(P<0.05)。4)与HD组相比,HD+LA组肉鸡血清钙含量和ALP活性显著升高(P<0.05),血清CORT含量以及NOX和MPO活性显著下降(P<0.05);采食频次和时间占比显著增加(P<0.05),饮水频次和趴卧时间占比显著降低(P<0.05);胫骨质量以及钙、磷含量与股骨密度、强度和钙含量显著上升(P<0.05)。综上所述,高饲养密度致使肉鸡发生氧化应激,改变了肉鸡行为,并使肉鸡骨骼质量下降,而饲粮中添加LA能在一定程度上缓解高饲养密度对肉鸡造成的损伤。     

油茶皂苷的制备及提纯工艺研究

为研究油茶皂苷的制备及提纯工艺,为中试化生产提供科学的技术支撑,通过正交试验得到最佳工艺为乙醇体积分数75%,料液比1∶2,提取时间120 min,提取温度60℃,得到纯度为55.65%的油茶皂苷粗品,得率16.12%。油茶皂苷粗品配置成质量分数3%水溶液,加入10%体积的1%壳聚糖絮凝剂絮,经凝沉、淀除去杂质,澄清液经AB-8型大孔树脂吸附90 min,用质量分数2%氢氧化钠溶液洗脱杂质,再用体积分数70%乙醇溶液洗脱油茶皂苷,将洗脱液旋转蒸发,除去乙醇并烘干,即得到纯度85.36%的油茶皂苷。     

甘蔗热带种金属硫蛋白家族基因的克隆及响应重金属胁迫的表达分析

金属硫蛋白是一类富含巯基的低分子量蛋白,在植物的重金属解毒及细胞氧化还原调控等方面起重要的作用。本研究以甘蔗热带种Badila组培苗为材料,分别测定了其在CdCl2、ZnSO4和CuCl2水溶液培养条件下地上部和地下部的重金属含量,结果显示其对上述3种重金属有较强的耐受与富集能力。继而克隆了ScMT1(登录号为KJ504373)、ScMT2-1-5(登录号为MH191346)和ScMT3(登录号为KJ5043704)3个金属硫蛋白家族基因,它们分别属于植物MT亚家族中的MT1、MT2和MT3型基因。ScMT1含有1个内含子和2个外显子,开放阅读框(Open Reading Frame,ORF)长228 bp,编码75个氨基酸;ScMT2-1-5含有2个内含子和3个外显子,ORF长246 bp,编码81个氨基酸;ScMT3含有1个内含子和2个外显子,ORF长198 bp,编码65个氨基酸。RT-qPCR显示,Cd2+胁迫下,在甘蔗地上部和地下部,ScMT2-1-5均连续显著上调表达,而ScMT1的上调应答出现延迟。ScMT3在地上部的上调应答出现延迟,在地下部呈“扬-抑”趋势,提示甘蔗响应Cd^2+胁迫过程中ScMT2-1-5起更积极的作用,ScMT1参与胁迫后期的分子响应,而ScMT3不起主导作用。Cu^2+胁迫下,地上部ScMT1连续显著上调表达,ScMT2-1-5和ScMT3呈总体上调的表达趋势;地下部,ScMT1和ScMT2-1-5的上调表答均出现延迟,仅在胁迫后期显著上调表达,而ScMT3仅在胁迫前期显著上调表达。该结果提示了ScMT1、ScMT2-1-5和ScMT3在Cu2+胁迫响应过程中的协作关系,三者共同参与了地上部的胁迫响应,其中ScMT1起更积极的作用;此外三者还先后参与了地下部对Cu^2+胁迫的分子响应。Zn^2+胁迫下,ScMT1和ScMT3分别仅在地上部和地下部显著上调表达;ScMT2-1-5在地上部和地下部均呈“扬-抑”的应答趋势;提示了在甘蔗响应Cd^2+胁迫应答过程中ScMT1和ScMT3分别在地上部和地下部起主要作用,ScMT2-1-5参与了胁迫前期的分子响应。ScMT1、ScMT2-1-5和ScMT3在甘蔗不同组织中及在重金属(Cd^2+、Zn^2+或Cu^2+)不同累积水平下呈现出相似或互补的应答特性,提示上述甘蔗MT家族不同成员在重金属解毒及细胞氧化还原调控等方面产生了功能分化,且三者在应对过量Cd^2+、Zn^2+或Cu^2+对甘蔗组织造成伤害的过程中存在时空上的协同作用。该研究为深入理解多倍体植物甘蔗中MT家族各成员基因在重金属耐受过程中的协同作用机制奠定了基础。     

Role of PERK pathway in morphine-induced myocardial protection of H9c2 cells

AIM: To explore whether morphine protects oxidative stress-damaged myocardial cells by inhibiting the PERK pathway to reduce endoplasmic reticulum stress and prevent mitochondrial permeability transition pore (mPTP) opening. METHODS: Rat myocardial H9c2 cells were cultured to establish an oxidative stress model, and then randomly divided into control group, H₂O₂ group, H₂O₂+morphine group, H₂O₂+morphine+PERK pathway inhibitor GSK2656157 group, morphine group and GSK2656157 group. Immunohistochemical method was used to detect the effects of morphine on expression of glucose-regulated protein (GRP) 78 and GRP94 induced by oxidative stress. The protein levels of PERK signaling pathway-related molecules were determined by Western blot. Confocal microscopy was used to observe the effects of morphine on mPTP opening and endoplasmic reticulum induced by oxidative stress. Cellular toxicity was detected by lactate dehydrogenase (LDH) kit and cell viability was measured by MTT assay. RESULTS: Compared with control group, GRP78 and GRP94 proteins in H₂O₂ group were strongly expressed, and the brown-yellow particles were significantly increased, but morphine significantly inhibited this process. Compared with control group, the phosphorylation of PERK was significantly reduced with GSK2656157 treatment at different concentrations, among which 2 μmol/L had the most significant effect (P < 0.05). Oxidative stress significantly increased the protein levels of GRP78, GRP94, p-PERK and CHOP, but significantly decreased p-GSK-3β level. These changes were inhibited by morphine, and the effects of morphine were further enhanced by GSK2656157 (P < 0.05). Compared with control group, oxidative stress significantly reduced the fluorescence intensity of mitochondrial TMRE and ER-Tracker Red. Morphine significantly inhibited this effect even when mitochondrial membrane potential was reduced, mPTP was open, and endoplasmic reticulum was damaged, while GSK2656157 further enhanced the effect of morphine (P < 0.05). Compared with control group, H₂O₂ significantly increased cellular toxicity and decreased the cell viability. Morphine inhibited this effect and GSK2656157 significantly enhanced the effect of morphine (P < 0.05). CONCLUSION: Morphine protects cardiac H9c2 cells under oxidative condition by inhibiting endoplasmic reticulum stress through PERK pathway and preventing the mPTP opening via GSK-3β inactivation.     

Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.     

Pyroptosis mediates high glucose-induced inflammation and injury in MC3T3-E1 osteoblasts

AIM To investigate whether pyroptosis contributes to the inflammation and injury in mouse embryonic osteoblastic cell line MC3T3-E1 induced by high glucose (HG; 45 mmol/L glucose). METHODS The cell viability was measured by CCK-8 assay. The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 (CASP1) were determined by Western blot. The secretion levels of interleukin-18 (IL-18) and IL-1β were measured by ELISA. The intracellular level of reactive oxygen species (ROS) was detected by 2',7'-dichlorodihydrofluorescein diacetate staining followed by photofluorography. Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorography. The alkaline phosphatase (ALP) activity was determined using the ALP kit, and the number of mineralized nodules was detected by alizarin red S staining. RESULTS After the MC3T3-E1 osteoblasts were treated with HG for 24 h, the protein expression levels of NLRP3 and CASP1, and the secretion levels of IL-18 and IL-1β were significantly increased. The decrease in cell viability, and the increases in ROS generation and MMP loss were also observed. Moreover, the differentiation and mineralization of MC3T3-E1 osteoblasts were inhibited, evidenced by decreases in both ALP activity and mineralized nodule number. Knockdown of CASP1 by siRNA attenuated the HG-induced osteoblast inflammation and injury mentioned above. CONCLUSION Pyroptosis mediates HG-induced inflammation and injury in MC3T3-E1 osteoblasts.