基于GF-6卫星影像多特征优选的酿酒葡萄精准识别

多源遥感信息和特征优选是提高农作物识别精度的重要支撑，高分六号（GF-6）卫星作为首次引入红边波段的国产卫星，其丰富的光谱信息为作物识别提供了新的思路和解决途径。该研究基于宁夏回族自治区银川市永宁县2018年6月-2019年3月的GF-6数据，充分利用红边优势提取光谱特征、纹理特征和植被指数特征，构建多种特征组合方案，并根据随机森林算法对特征重要性进行度量，选取最优特征组合对酿酒葡萄进行精准识别。结果表明，与单一特征相比，多源遥感特征的增加显著改善了酿酒葡萄分类效果，其中，植被指数贡献程度最大，光谱特征次之；基于随机森林的优选特征组合分类效果最佳，其中，总体分类精度为94.15%，酿酒葡萄用户精度为94.23%，制图精度为92.59%；以实地调查的4个酒庄为验证区，将酿酒葡萄提取结果与统计数据进行对比，面积相对精度均在70%以上，其中优选特征结果相对精度在90%以上，研究结果将为国产卫星红边波段在植被分类和识别方面的应用提供数据参考。     

m6A甲基化在鸡肌肉生长发育中的表达研究

试验旨在研究RNA m6A修饰相关基因去甲基化酶Alk B同源蛋白5（Alk B homologue 5，ALKBH5）、去甲基化酶肥胖相关蛋白（fat mass and obesity-associated protein，FTO）、甲基转移酶样蛋白3（methyltransferase like 3，METTL3）、甲基转移酶样蛋白14（methyltransferase like 14，METTL14）和成肾细胞瘤1-结合蛋白（Wilms’tumor 1-associating protein，WTAP）在鸡骨骼肌发育过程中的表达，分析其与骨骼肌m6A甲基化水平的相关性。首先，利用实时荧光定量PCR技术检测m6A甲基化相关基因在金茅花鸡12（E12）、14（E14）、16（E16）、18（E18）胚龄和1日龄腿肌和胸肌组织中mRNA表达水平，以及其在鸡成肌细胞50%、100%增殖期和1、2、3、4、5 d分化期的mRNA表达水平；随后，利用m6A甲基化试剂盒检测金茅花鸡E12和1日龄腿肌和胸肌组织中m6A甲基化修饰水平，与m6A甲基化相关基因表达水平进行相关性分析。结果显示，m6A去甲基化基因ALKBH5和FTO mRNA表达水平在骨骼肌发育过程中显著上调（P<0.05），即在E12、E14低表达，E16、E18逐渐上调，1日龄达到最高。m6A甲基化写入基因METTL14、METTL3和WTAP mRNA表达水平在E12、E14、E16逐渐上升，E18下降，随后至1日龄表达量回升。在细胞增殖过程中，ALKBH5、FTO、METTL14、METTL3和WTAP基因表达均上调；在细胞分化过程中ALKBH5和FTO基因表达水平显著上调（P<0.05），在分化第5天达到最高。METTL14、METTL3和WTAP基因mRNA表达水平在细胞诱导分化的1、2、3、4 d表达量呈下降趋势，而在诱导分化的第5天有所回升。甲基化水平检测结果显示，腿肌和胸肌m6A甲基化水平变化趋势一致，均在胚胎发育过程中显著下降（P<0.05），至1日龄达到最低。相关性分析结果显示，鸡骨骼肌RNA m6A甲基化水平与m6A去甲基化修饰基因ALKBH5、FTO mRNA表达水平呈显著负相关（P<0.05）。综合以上试验结果，推测m6A甲基化修饰与鸡骨骼肌发育相关，而去甲基化基因ALKBH5、FTO可能通过调控RNA m6A甲基化水平，影响鸡骨骼肌发育。本研究结果为进一步研究m6A甲基化修饰调控鸡骨骼肌生长发育的功能和分子机制提供理论依据。     

棉花根系和叶片质外体汁液分离方法的改进

质外体汁液(apoplastic washing fluid, AWF)在植物生长发育和抵抗生物及非生物逆境方面发挥着重要作用。目前普遍采用真空渗透离心法提取质外体汁液,但具体提取流程和条件却因植物培养条件、种类和器官等不同而不同。本试验以营养液培养的棉花幼苗为材料,改进了棉花根系和叶片AWF分离的取样方法、渗透条件和离心参数等。结果表明,相对于通常的非整体取样,改良后整体取样简单易操作且显著降低了质外体与共质体的苹果酸脱氢酶(malate dehydrogenase, MDH)活性比值,更适宜开展质外体汁液组分研究。根据真空渗透鲜重增加和AWF稀释因子,进一步证明根系不用真空渗透;而叶片需真空渗透,适宜真空强度/时间为-60 kPa/1min,真空后恢复到正常压强约110s。证明叶片颜色变深面积可作为判定真空渗透强度或时间是否适宜的一种简易可行方法。最后,AWF体积、质外体与共质体的可溶性蛋白含量比值和MDH活性比值综合分析表明,棉花根系适宜离心力大小/时间长度为800×g/10～20 min,叶片为400×g/5 min。本改进方法为棉花AWF组分,如蛋白质组学和代谢组学等研究结果的准确性和可靠性奠定了基础,为其他作物AWF分离方法的优化提供了参考。     

夏训对优秀皮划艇运动员铁调素等铁营养状况以及IL-6的影响

目的：探讨夏训对优秀运动员铁调素等铁营养状况以及IL-6的影响。方法：广东省优秀皮划艇运动员26人(男15人，女11人)参加测试，调查夏训前和夏训后血红蛋白、铁调素、可溶性转铁蛋白受体、血清铁蛋白、可溶性转铁蛋白受体/log铁蛋白、血清铁、总铁结合力、转铁蛋白饱和度以及白介素6变化情况。结果：夏训后，男运动员组铁调素和可溶性转铁蛋白受体极显著上升(P<0.01)、血清铁蛋白极显著下降(P<0.01)，可溶性转铁蛋白受体/log铁蛋白显著上升(P<0.05)，总铁结合力显著下降(P<0.05)，白介素6极显著上升(P<0.01)。女运动员组血红蛋白显著降低(P<0.05)，铁调素和可溶性转铁蛋白受体极显著上升(P<0.01)，血清铁蛋白极显著下降(P<0.01)，可溶性转铁蛋白受体/log铁蛋白极显著上升(P<0.01)，白介素6极显著上升(P<0.01)。结论：夏训可引起运动员炎症因子升高、铁调素升高，导致功能铁不足；铁调素介导的铁代谢紊乱可能是运动性低血色素发生的重要原因。     

艾纳香脱氢酶基因家族的生物信息学分析

采用RT-PCR和RACE（Rapid amplification of cDNA ends）技术,从艾纳香（Blumea balsamifera (L.) DC.）叶片中克隆到4条编码艾纳香脱氢酶（BbADH1、BbADH2、BbADH3、BbADH4）基因的cDNA序列,并对4条核苷酸及其编码的氨基酸序列进行生物信息学分析。结果显示:4条艾纳香脱氢酶序列开放阅读框均在900 bp左右,蛋白质等电点（pI）值在5.0~9.0之间,含量最多的氨基酸为亮氨酸（Leu）,最少的为色氨酸（Try）,具有明显的疏水区和亲水区,N端未发现信号肽,且无跨膜区;同源性比对结果显示,艾纳香BbADH蛋白与其他植物中ADH蛋白具有高度的相似性,且具有脱氢酶的特征功能域;系统发育分析表明,艾纳香（B. balsamifera (L.) DC.）BbADH1和BbADH3同处于一个分支,且与胡杨（Populus euphratica Oliv.）PeADH物种亲缘关系较近,而艾纳香BbADH4和BbADH2处于不同分支,且分别与葡萄（Vitis vinifera）VvADH和烟草（Nicotiana tabacum）NtADH物种亲缘关系较近。     

云南和广东省流行性出血病病毒血清6型毒株(EHDV-6)的分离与遗传特性

【目的】分离流行于我国南方地区的流行性出血病病毒血清6型毒株(EHDV-6),掌握分离毒株的遗传特征。【方法】对设立于我国云南和广东省的哨兵动物定期采血,进行EHDV感染情况监测与病毒分离培养。通过血清中和试验确定分离EHDV的血清型,采用一步法RT-PCR对分离的EHDV-6毒株基因节段2、3、6(Seg-2、-3、-6)进行扩增与序列分析。【结果】2012—2016年,从云南省和广东省的哨兵动物中分离出25株EHDV毒株,其中,11株为EHDV-6型。中国EHDV-6型毒株的Seg-2、-3与-6均属Eastern型,核酸序列相似性分别为99.1%、98.9%和98.8%,在系统发生树上分别聚为1个独立的中国支系。在日本引起疫病暴发的EHDV-6型毒株与中国EHDV-6型毒株具有最近的亲缘关系。日本EHDV-6型毒株在系统发生树上处于中国支系内部,Seg-2、Seg-3与中国毒株的核酸序列相似性分别为98.5%和93.9%。【结论】云南和广东省流行的EHDV-6型毒株核酸与氨基酸序列高度相似;日本和中国的EHDV-6型毒株具有最近的亲缘关系,提示中日之间可能存在EHDV的流动。研究结果为进一步开展我国EHDV-6型毒株流行病学分析、致病性、病原学诊断与疫苗制备等研究提供了基础。     

Expression and mechanisms of hsa_circ_087631 in patients with primary biliary cholangitis

AIM To explore the expression and mechanisms of circular RNA hsa_circ_087631 in the patients with primary biliary cholangitis （PBC）. METHODS RT-qPCR was used to detect the expression of hsa_circ_087631 in PBC patients and healthy controls. Hsa-miR-346-overexpressing lentiviral vector pLenti-EF1a-EGFP-F2A-Puro-CMV-MCS was constructed and transfected into human acute T cell leukemia Jurkat cells， and then the expression levels of hsa_circ_087631， Bcl-6 mRNA and interleukin-21 （IL-21） mRNA were detected by RT-qPCR. Dual-luciferase reporter assay was performed to identify the interactions between hsa_circ_087631 and hsa-miR-346. RESULTS The expression of hsa_circ_087631 in the PBC patients was significantly increased compared with the healthy subjects. Hsa-miR-346-overexpressing lentiviral vector pLenti-EF1a-EGFP-F2A-Puro-CMV-MCS was successfully constructed. The expression of hsa-miR-346 was significantly increased after the hsa-miR-346-overexpressing lentiviral vector was transfected into the Jurkat cells， while the expression levels of hsa_circ_0087631， Bcl-6 mRNA and IL-21 mRNA were significantly decreased. After wild-type or mutant hsa_circ_087631 vector and hsa-miR-346 mimics were transfected into 293T cells， the results of dual-luciferase reporter assay showed that hsa-miR-346 significantly decreased the luciferase activity of wild-type hsa_circ_087631 （P<0.01）， but the regulation did not change significantly after mutation of the predicted binding site. CONCLUSION Peripheral blood hsa_circ_087631 level is elevated in the PBC patients. The hsa_circ_087631/hsa-miR-346/Bcl-6 signaling may take effect in human T cells. Hsa-miR-346 significantly reduces the expression of hsa_circ_087631， but it may not be regulated by predicted binding sites.     

Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

Uneven ripening in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) berries in India is linked to seed viability

Uneven ripening (UR) is a physiological disorder of unknown origin in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) leading to wine of inferior quality. A preliminary study found wide variations in total dehydrogenase activity (TDH) of seeds from unevenly ripe berries. In our experiments, gibberellin (GA₃) applied to young grapes increased seed TDH activity and reduced the incidence of uneven ripening to 2% compared with 35% in the control. In contrast, paclobutrazol (PBZ) decreased TDH activity and increased the incidence of the disorder to 58%. GA₃-treated berries had higher concentrations of sucrose and TDH activity in seed representing mature seeds with high viability. In contrast, PBZ-treated and control berries had higher concentrations of glucose and lower TDH activity, indicating immature seeds with low viability. These results suggested that competition among developing berries can lead to differences in seed gibberellin content, seed viability and the rate of berry growth resulting in green, purple, and black berries at harvest. The study established the role of seed viability in uneven ripening and demonstrated that the incidence of the disorder is reduced by the application of GA₃ to immature berries.     

黄瓜低霜霉威残留性相关基因SDH克隆及表达

通过Solexa技术和比较基因组学方法鉴定霜霉威胁迫下黄瓜果实差异表达基因,筛选黄瓜低霜霉威残留性相关基因SDH。研究利用PCR克隆获得SDH基因全长,命名为Cs SDH。构建亚细胞定位表达载体,利用农杆菌浸润法注射烟草叶片后激光共聚焦显微镜下观察;分别用荧光定量RT-PCR方法和酶联免疫法研究高、低农残品系D9320和D0351中Cs SDH基因在农药霜霉威处理后,不同时间点、不同组织部位时空表达特征和琥珀酸脱氢酶活性。结果表明,Cs SDH基因在烟草叶片细胞中被定位在细胞膜上。黄瓜低农残品种D0351中,Cs SDH基因在果实受霜霉威胁迫后反应迅速,主要在处理前期响应表达强烈,琥珀酸脱氢酶活性显著增加,且该基因表达量显著高于其在高农残品种D9320中表达量。Cs SDH果、叶中表达量较茎中高,且各组织部位表达量高低顺序稳定,为叶果茎。黄瓜高农残品种D9320中,Cs SDH基因表达量在0.5、1、3、12 h呈现上调表达,6、9 h基因表达量差异不显著,48 h出现上调表达高峰。说明克隆得到的Cs SDH基因属农药处理前期响应基因,且该基因可能在降低黄瓜农药残留过程中发挥重要作用。试验通过研究黄瓜琥珀酸脱氢酶基因(SDH)在霜霉威胁迫下表达情况,为挖掘黄瓜低农药残留性功能基因以及探明其作用分子机制奠定基础。