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.     

基于GIS与RS的北方防沙屏障带生态系统格局演变

研究北方防沙屏障带生态系统格局演变特征,对加强屏障带建设、改善生态环境具有重要意义。以北方防沙屏障带为研究区,根据2005、2010、2015年MODIS遥感影像,将北方防沙屏障带生态系统类型划分为森林、草地、湿地、农田、城镇、荒漠和裸地7类。采用空间统计、转移矩阵、景观指数分析法、PNTIL模型等方法,对北方防沙屏障带2005—2015年生态系统类型时空演变特征进行分析。结果表明,2005—2015年北方防沙屏障带生态系统整体呈稳定状态;从分屏障带看,内蒙古防沙屏障带荒漠面积明显减少,塔里木防沙屏障带森林面积迅速增加,而河西走廊防沙屏障带草地面积明显增加;从正/逆转换方向来看,屏障带内农田、城镇、荒漠及裸地生态系统正向转换率高于逆向转换率。10年间,在景观水平上,北方防沙屏障带斑块数量、斑块密度呈减少趋势;在类型水平上荒漠景观整体呈现破碎化萎缩的趋势特征,森林、草地向形状简单化的方向发展。研究表明,2005—2015年,北方防沙屏障带生态总体向好,治沙效果明显,但局部仍面临较大压力。     

高致病性猪繁殖与呼吸综合征病毒株JXA1反向遗传平台的构建

【目的】为高致病性猪繁殖与呼吸综合征病毒(Highly pathogenic porcine reproductive and respiratory syndrome virus,HP-PRRSV)的结构功能和致病机理的研究奠定基础。【方法】运用反向遗传技术将HP-PRRSV JXA1株的全基因组分段克隆至改造过的低拷贝载体pOKq上,并在病毒基因组两端分别添加CMV启动子和BGH终止信号肽以及在病毒全基因组第510位核苷酸突变引入Fse I酶切位点,作为遗传标记位点。采取基于DNA-launched途径进行病毒拯救,并对拯救的病毒进行生物学特性分析。【结果】构建的PRRSV JXA1毒株的全长cDNA克隆具有感染性;成功拯救了病毒,命名为rJXA1;成功引入了拯救病毒的遗传标记;拯救病毒与亲本病毒的生长曲线相似,二者达到最高滴度的时间均为感染后72 h。【结论】成功构建了JXA1株反向遗传平台,为进一步研究HP-PRRSV的致病机理、基因功能以及新型疫苗研发奠定了基础。     

行星轮系水稻钵苗移栽机构正反求设计方法研究

为了解决现有行星轮系水稻钵苗移栽机构较难同时具备理想的移栽轨迹和姿态的问题,提出了正向设计与局部轨迹微调的反求设计相结合的设计方法。以非圆齿轮行星轮系水稻钵苗移栽机构的设计为例,阐述该方法的实现过程。建立行星轮系水稻钵苗移栽机构设计要求,并分析其工作原理,在前期正向设计的基础上,局部调整其移栽静轨迹,通过反求设计进一步优化设计移栽机构。进行局部反求设计的运动学分析和运动学建模,基于Matlab平台开发计算机辅助设计软件,通过人机交互的方式得到移栽机构的机构参数,使其不仅满足移栽臂移栽的姿态要求,同时获得更理想的移栽工作轨迹。根据最终得到的机构参数设计移栽机构结构,完成三维建模、虚拟仿真试验,加工、装配移栽机构物理样机,完成了高速摄像运动学试验。将移栽机构理论计算、虚拟仿真、样机试验得到的移栽静轨迹比较,同时逐一将轨迹相关参数、取秧角、推秧角、角度差等设计目标参数与相应数值化目标进行对比,结果显示,均满足要求,验证了设计方法的正确性。与以往单一的正向设计、反求设计方法相比,该方法设计的移栽机构不仅具有更优的移栽工作轨迹,也可满足较好的移栽姿态要求。     

土地确权对农户地权稳定性感知的影响——基于村庄内土地不平等视角的分析

稳定的农地产权能够激励农户农地投资,有助于促进农地资源可持续利用和农业可持续发展。土地确权是稳定农地产权的重要正式制度安排,即稳定地权是新一轮土地确权承载的重要功能。基于诱致性制度变迁理论和社会心理学理论,以黄土高原苹果优势区为研究对象,利用实地调研数据,采用Logit和Ologit模型,从村庄内家庭土地不平等视角分析土地确权对农户地权稳定性感知的影响及其作用机制,探讨新一轮农地确权的地权稳定效应。结果表明,在研究区域,33.79%的农户已完成新一轮土地确权颁证,62.64%的农户认为研究地块在未来不会被重新调整,即感知的地权稳定性水平较高。新一轮土地确权显著强化了农户的地权稳定性感知,且在控制村庄内土地不平等的情境中具有更高的强化效果;村庄内家庭土地不平等对农户地权稳定性感知具有显著的弱化效应,且土地不平等在土地确权对农户地权稳定性感知的影响中具有负向调节作用。因此,在"三权分置"产权制度改革中,政府应重视规范土地确权程序,加强产权制度改革宣传和信息披露,同时,注重以政策调整和市场配置相结合的方式,降低土地不平等造成的负面影响,强化土地产权制度改革的政策效果。     

鞭毛蛋白及鸡GM-CSF重组新城疫病毒对鸡的免疫增强作用

为评价鼠伤寒沙门氏菌鞭毛蛋白(FliC)以及鸡GM-CSF(chGM-CSF)对新城疫病毒(NDV)弱毒疫苗的免疫增强效果,本实验通过PCR扩增fliC,并合成chGM-CSF编码基因,分别构建重组质粒pBrClone30-fliC和pBrClone30-GM-CSF,拯救获得重组病毒rClone30-fliC和rClone30-GM-CSF。RT-PCR和ELISA检测结果表明:fliC和chGM-CSF基因正确插入重组病毒,并获得稳定表达;重组病毒免疫7 d后重组NDV HI抗体效价检测显示,实验组平均HI效价分别达到4.5 log2和4.8 log2,而r Clone30组和空白组HI效价仅为3 log2和1 log2;免疫7 d后攻毒,空白组SPF鸡均死亡,rClone30组有两例发病,实验组SPF鸡全部存活并且无任何临床症状。本研究结果表明,FliC和chGM-CSF作为佐剂能够有效提高弱毒疫苗的免疫效果。     

PA-X基因的长度变化对H3N2犬流感病毒复制能力和致病性的影响

H3N2犬流感病毒(canine influenza virus, CIV)已在中国多地的犬群中流行,是禽流感跨宿主感染并形成新分支的近期案例。研究表明,PA-X基因与甲型流感病毒适应新宿主的能力相关,且其长度能够影响甲型流感病毒的复制及致病能力。为了解PA-X基因的长度变化对H3N2 CIV复制能力及致病力的影响,本研究利用H3N2 CIV的8质粒操作系统,拯救了三株重组H3N2 CIV毒株：PA-X基因表达大小为232个氨基酸多肽的亲本病毒CIV＿PA-X＿232;对PA编码区第191、192位氨基酸的密码子进行改造,PA-X基因不表达蛋白的重组病毒CIV＿PA-X＿Knock;对PA+1编码区第232位氨基酸进行突变,PA-X基因表达大小为252个氨基酸多肽的重组病毒CIV＿PA-X＿252。通过比较3株重组病毒的聚合酶活性,在MDCK细胞中的复制效率及对小鼠致病性的差异,来评价表达不同长度的PA-X基因对H3N2 CIV的影响。结果显示,CIV＿PA-X＿252和CIV＿PA-X＿Knock的聚合酶活性显著(P<0.05)高于CIV＿PA-X＿232,且CIV＿PA-X＿...     

TILLING技术及其在农作物中的应用现状

TILLING技术是一种全新的反向遗传学研究方法。文章简要介绍了TILLING技术的原理、操作流程、技术要点及其特点,并总结了TILLING技术在麦类、玉米及水稻研究中的应用现状。     

Evidence of sedimentation inequality along riparian areas colonised by Impatiens glandulifera (Himalayan balsam)

Soil loss from riparian areas supporting the annual invasive weed, Impatiens glandulifera (Himalayan balsam), was measured and compared with equivalent values recorded at nearby, topographically similar areas supporting perennial vegetation over a cumulative seven-year period, along sections of two separate river systems; one in Switzerland, and one in the UK. Soil loss from colonised locations was significantly greater than from reference locations in four of the seven measurement periods. Despite contrasting results, standard deviations, based on soil losses and gains, were predominantly higher for colonised areas at both rivers over most monitoring periods. These findings indicated that areas colonised by Himalayan balsam experience higher sediment flux in comparison with areas free of invasion. Here, we test those original interpretations by reinterrogating the datasets using a more robust analysis of inequality. Nine datasets were tested, five of which (i.e. 56%) showed that sediment flux was significantly greater at Himalayan balsam-invaded areas than at reference areas. Three datasets showed no difference in sediment flux between invaded and reference areas (33%), and one (11%) showed higher sediment flux at reference areas. Most results uphold our original interpretations and support our hypothesis that hydrochory probably dictates where colonisation initially occurs, by depositing Himalayan balsam seeds in slack or depressional areas along river margins. Once Himalayan balsam becomes established and sufficient perennial vegetation is displaced, seasonal die-off and depleted vegetation cover may increase the risk that some areas will experience significantly higher sediment flux.