Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.     

成都旅游节庆活动同质化问题及其对策研究

从目前各地节庆项目的举办情况出发,分析了成都发展旅游节庆项目的优势,探讨了部分节庆活动在主题选择、活动内容、操作方式等方面存在的问题,并从挖掘文化、注重调研、整合资源、适度引导和加强宣传等方面提出旅游节庆活动进一步发展的思路。     

辐射诱变对不同遗传背景大豆杂交后代蛋白质和脂肪含量改良

高蛋白材料匮乏及选育方法单一是辽宁省高蛋白大豆育种中的两个突出问题。为探索高蛋白大豆种质资源创新有效途径,本研究以辽宁省农业科学院选育的辽04Q086、辽08020、辽05086和辽06Q003高蛋白品系为母本,与河北省沧州市农林科学院选育的高蛋白品种沧豆10号进行有性杂交,并对亲本及杂交F_2代进行不同剂量的辐射诱变处理,分析了不同剂量辐射诱变对不同受体的籽粒蛋白质和脂肪含量的改良效果。结果表明:4个大豆品系与沧豆10号杂交衍生的F_3代植株中,低蛋白高脂肪类型所占比例最高,达到了45.0%。4个大豆品系经辐射诱变(~(60)Coγ-14000和~(60)Coγ-16000)衍生的M_1代中,低蛋白高脂肪类型所占比例也分别达到42.5%和40.0%。然而,以杂交F_2代种子为受体进行不同剂量辐射诱变衍生的M_1代植株中,高蛋白低脂肪类型所占比例最高,分别为35.0%和27.5%,蛋白质含量最高达47.0%。本研究表明,以不同遗传背景的高蛋白大豆杂交,并结合辐射诱变选育高蛋白材料是可行的。     

茶树蛋白激酶基因CsCIPK的克隆与表达特性分析

以茶树品种‘龙井43’作为材料,利用RT-PCR方法,从茶树的cDNA中克隆得到1个编码蛋白激酶的基因,命名为CsCIPK。序列分析表明,CsCIPK开放阅读框长度为1 341 bp,编码446个氨基酸,蛋白质分子量为414234。蛋白功能域预测和多重对比显示,CsCIPK蛋白含有1个保守的N端激酶结构域和1个相对不保守的C端调节结构域,即丝氨酸/苏氨酸激酶结构域和NAF结构域。理化性质、亲/疏水性、无序化分析显示,CsCIPK属于疏水性蛋白,理论等电点为7.04,有4段无序化区域,其二级结构分析显示主要由α螺旋、不规则卷曲组成。通过实时荧光定量PCR对‘龙井43’和‘安吉白茶’中的CsCIPK表达特性进行分析。结果显示‘龙井43’中CsCIPK的相对表达量在高温、干旱及盐处理4 h、低温处理24 h时达到最高。‘安吉白茶’中CsCIPK的相对表达量在高温及盐处理4 h、低温及干旱处理1h时达到最高。CsCIPK在‘龙井43’的根中,‘安吉白茶’茎中表达量最高。不同浓度的GA和IBA处理‘龙井43’茶苗,结果显示0.2 mmol·L-1 GA处理后,CsCIPK表达量先升高后下降,6 d时处理组为对照组的62倍;0.6 mmol·L-1 IBA处理后,CsCIPK的表达量在3 d时显著高于对照组;不同浓度GA和IBA处理后,9 d时CsCIPK表达量均显著低于对照。     

1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

基于形态学及多基因位点的枸杞褐斑病病原鉴定

枸杞Lycium barbarum Mill.是一种药食两用的名贵中药材,褐斑病是2010年在甘肃省枸杞种植区发生的由无性态类真菌新种枸杞小黑梨孢Stigmella lycii引起的新病害,据2015年-2018年调查,该病害已扩展至甘肃省各枸杞种植区,常年发病率45%～65%,严重度2～3级。本研究首次对枸杞小黑梨孢有性态形态进行描述,并结合分子生物学方法,确定该病病原菌的系统发育地位,为该病的防治提供一定的理论依据。枸杞褐斑病菌在离体培养条件下和越冬病叶上均可形成有性态。子囊壳球形至亚球形,大小185.6μm×176.6μm,有喙,喙的大小为(35.7～53.6)μm×(32.0～53.55)μm;子囊袋状,大小(103.2～165.7)μm×(15.7～22.4)μm,含8个子囊孢子;子囊孢子砖格状,大小(25.9～36.5)μm×(9.4～14.1)μm,平均31.2μm×12.3μm,具(4～)6～7个纵隔和1～3个横隔。通过ITS、LSU、RPB2和EF1-α多基因位点联合构建系统发育树,确定该菌为子囊菌门格孢腔菌目格孢腔菌科真菌。秋末冬初清洁田园,减少来年初侵染源,是有效防治枸杞褐斑病的关键措施。     

智能氮肥在干湿交替灌溉下对水稻产量与水分利用率的影响

为探究智能氮肥(pH响应性控释氮肥)在干湿交替灌溉模式下的适宜性,设置3种程度的干湿交替灌溉W1(灌水下限100％饱和含水率)、W2(灌水下限90％饱和含水率)、W3(灌水下限70％饱和含水率),3种施肥形式智能氮肥(SF)、普通无机氮肥(MF)以及不施氮肥(NF)的桶栽试验,分析其对水稻产量及水分利用效率的影响.结果表明:SF×W1处理下下产量最高,为64.84 g/桶,高出常规氮肥21.8％(W1)、13.9％(W2).SF×W1处理下水分生产效最高,为3.17 kg/m3,SF×W2处理下灌水生产率最高,为1.20 kg/m3.智能氮肥在与适度干湿交替灌溉结合下,能达到节水、增产的目的,是实现水稻节水、高产、控肥的有效途径.     

优化提取裸花紫珠总黄酮含量及对表皮伤口易感染菌抑菌性的研究

为了选出裸花紫珠总黄酮提取的最优方法,比较了ASE、超声、微波萃取和多功能浓缩4种提取方法在裸花紫珠总黄酮提取率上的差异,并利用微孔刃天青法测定总黄酮对表皮易感染菌的MIC值。结果发现ASE法总黄酮提取率最高,约为1.26%,其次为超声和微波萃取法,总黄酮提取率分别约为1.03%和0.98%,多功能浓缩法提取率最低,约为0.36%,且耗时最长。而且研究发现,ASE的MIC值最低。最终得出ASE提取效率最高,且提取物有较好的抑菌性,是裸花紫珠总黄酮提取的最优方法。     

水稻对稻飞虱抵御机制研究

为更好地利用水稻自身抵御能力遏制稻飞虱为害,综述了水稻抵御稻飞虱的物理、生化效应和机制的研究现状。水稻抵御稻飞虱的物理机制包括宏观的水稻叶脉间距、宽度,钩毛和纤细毛密度、长度,硅含量和微观的粗纤维、胼胝质含量及叶绿体膜稳定性;生化机制涉及水分、可溶性糖、叶绿素、游离氨基、酶物质活性及挥发物的含量。国内外虽广泛涉及水稻自身固有的物理结构及体内生理生化物质对稻飞虱的抵御效应研究,但其分子机理研究较为匮乏,尚需深入系统的研究。