MicroRNA-9-5p mediates sympathetic overactivity by targeting KCNN3 in paraventricular nucleus of rats with T2D

AIM To investigate whether microRNA-9-5p （miR-9-5p） mediates sympathetic overactivity by targeting KCNN3 （potassium intermediate/small conductance calcium-activated channel, subfamily N, member 3） gene,which encoded small-conductance calcium-activated potassium channel 3 （SK3） protein, in paraventricular nucleus （PVN） of rats with type 2 diabetes mellitus （T2D）. METHODS A rat model of T2D was established by high-fat diet combined with intraperitoneal injection of 30 mg/kg streptozotocin. The levels of miR-9-5p and KCNN3 mRNA in PVN were detected by real-time PCR. The relationship between KCNN3 and miR-9-5p was predicted by TargetScan. Recombinant adeno-associated virus （rAAV）-miR-9-5p or KCNN3 were bilaterally microinjected into the PVN to observe the changes in plasma glucose levels and sympathetic drive indicators. The number of FosB and SK3 positive cells was measured by immunofluorescence staining. The protein expression of SK3 was determined by Western blot. The relationship between KCNN3 and miR-9-5p were confirmed by cell transfection and dual-luciferase reporter assay. RESULTS Compared with the rats in diabetes control （DC） group, the blood glucose, sympathetic drive indexes and the level of miR-9-5p in PVN were significantly increased, while the SK3 expression in PVN was obviously reduced in the diabetes mellitus （DM） rats. After microinjecion of rAAV-miR-9-5p in PVN, the sympathetic drive indexes, blood glucose, and the number of FosB-positive cells were increased significantly, but the SK3 protein expression was significantly reduced （P<0.05）. However, up-regulation of KCNN3 in PVN had the opposite effect. These responses were obviously enhanced in DM rats compared with DC rats. The results of cell transfection and dual-luciferase reporter assay demonstrated that miR-9-5p bound to the 3’-UTR of KCNN3 and inhibit its expression. CONCLUSION miR-9-5p was up-regulated in PVN of the rats with T2D, and it may mediate sympathoexcitation by targeting KCNN3.     

白居易的园林情结及其对传统私家园林的影响

本文阐述了唐朝诗人白居易的园林情结,剖析其园林思想,并归纳他的“中隐”思想和朴实造园手法对传统私家园林产生的深远影响。     

土著B[a]P降解菌群的富集及最佳降解条件研究

为了获得能够降解苯并[a]芘(BaP)的微生物菌群,为生物修复多环芳烃污染的土壤提供菌种资源,本试验从长期石油污染的土壤中,富集了一个能够降解BaP的微生物菌群,并研究了其最佳降解条件,通过高通量测序研究了其群落结构。该菌群可以在15 d内将30 mg·L^-1的BaP降解33.34%。高通量测序的结果表明,该菌群主要由Bacillus、Zobellella、Gordonia和Rheinheimera组成,其中Bacillus是主要的降解菌。试验结果表明,该菌群的最佳降解条件为1%盐度,酵母浓度为80 mg·L^-1,pH值7.0。该菌群对芘、菲、荧蒽等多环芳烃也有一定的降解效果,其中荧蒽的降解效果最好,降解率达到99%。试验结果表明,该菌群在实际应用中具有很大的潜力。     

番茄不孕病毒BJ株系基因组测定与侵染性克隆

番茄不孕病毒(Tomato aspermy virus,TAV)可侵染包括藜科(Chenopodiaceae)、茄科(Solanaceae)等在内的24个双子叶家族和3个单子叶家族的100多种植物,是具有重要经济价值的植物病毒之一.为研究TAV BJ株系(Tomato aspermy virus,TAV-BJ)的基因组功能,本实验对TAV-BJ基因组克隆测序,并构建侵染性克隆.以TAV-BJ侵染心叶烟(ic otiana glutinosa)的总RNA为模板,RT-PCR获得其RNA2和RNA3;以TAV-BJ的dsRNA为模板,RT-PCR获得全长RNA1,目的片段PCR产物克隆测序获得TAV-BJ基因组全序列信息.RNA1全长3 409 nt,编码994个氨基酸的1a蛋白;RNA2全长3 023 nt,含2个开放阅读框(open reading frame,ORF),2a ORF编码829个氨基酸的2a蛋白,2b ORF编码78个氨基酸的2b蛋白;RNA3全长为2 216 nt,包含2个ORF,3a ORF编码247个氨基酸的3a蛋白,外壳蛋白(coat protein,CP)ORF 编码219个氨基酸的CP蛋白(TAV-BJ基因组RNA1、2和3 GenBank登录号分别为HQ424163,HQ424164和HQ424165).TAV-BJ基因组cDNA克隆体外转录成RNA并接种于心叶烟上,结果表明转录产物在寄主上的症状反应和TAV-BJ病毒粒子RNA的接种相一致,TAV-BJ基因组cDNA侵染性克隆具有活性.由TAV-BJ各个基因片段与缺失2b基因的黄瓜花叶病毒Fny株系(Cucumber mosaic virus,CMV-Fny△2b)构建的假重组病毒接种于心叶烟,结果显示TAV-BJ的RNA2和RNA3能恢复CMV-Fny△2b在寄主上症状反应.嵌合型RNA3F3aTcp和RNA3T3aFcp的症状反应结果表明,F1F2△2bRNA3 T3aFcp在寄主上产生花叶症状与F1 F2△2bT3相一致.本研究获得TAV-BJ的基因组序列,成功构建侵染性克隆,同时发现TAV-BJ的3a基因具有CMV-Fny的2b基因的某些功能.     

城市绿地土壤理化性质和Pb、Cd含量空间分布——以重庆市北碚区为例

城市绿地土壤作为城市生态系统的重要组成部分,对城市的可持续发展有重要的意义。为了更好地了解城市绿地土壤、充分发挥其功能,该文对重庆市北碚城区土壤容重、pH、有机质、CEC等理化性质和Cd、Pb含量的空间分布特征进行了分析。结果表明:西南部的土壤容重和pH值高于东北部;有机质整体偏高,且东北部高于西南部;西南部地区绿地土壤CEC含量略高于东北部;东北部地区土壤Cd和Pb含量高于西南部地区。绿地土壤在短时间内受到人为干预较为显著,但随着绿地土壤利用年限的增加,有向其所在地自然土壤特性发展的趋势。与土壤环境质量标准（GB15618-1995）相比,Pb、Cd含量均以二级标准为主,虽然其对人类和环境的危害性现在还没有体现出来,但随着时间推移,其危害性会逐渐体现出来,应引起我们的高度重视。     

蚯蚓（Eiseniafetida）细胞色素P450及抗氧化酶系对环境浓度苯并（a）芘的响应

以赤子爱胜蚓（Eiseniafetida）为供试生物,草甸棕壤为供试土壤,以蚯蚓微粒体细胞色素P450含量、抗氧化酶系以及谷胱甘肽转移酶活性为指标,进行了土壤中苯并（a）芘[B（a）P]暴露与酶活性的量-效关系研究。结果表明,在接近沈抚灌区实地污染状况B（a）P（0．1～2．0mg·kg-1）暴露下,第1、3、7以及第14d取样时,P450和SOD有较好的响应：SOD在第1、3d显著升高,而在第7、14d时降低;P450总体表现为低浓度B（a）P诱导下降低,高浓度下升高的趋势。CAT、POD以及GST的敏感性相对较差。比照其他4个指标,蚯蚓P450的敏感性更为优异,具有较好的应用前景。指标敏感性总体表现为：P450〉SOD〉CAT／POD〉GST。     

纳米ZnO对浮萍（Lemna minor L.）的生态毒性及其聚集性和溶解性分析

纳米颗粒的大量生产和应用增加了其环境释放风险,为此以不同浓度的纳米ZnO（0、1、10mg·L-1和50mg·L-1）处理浮萍（LemnaminorL.）7d,分析了纳米颗粒对植物D665/D665a值（叶绿素与脱镁叶绿素的比率）以及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、过氧化物酶（POD）和Na＋K＋-ATP酶（Na＋K＋-ATPase）活性的影响,并对纳米ZnO的聚集性与溶解性进行了测试。研究结果显示,浓度为50mg·L-1的纳米颗粒显著抑制D665/D665a值和Na＋K＋-ATPase活性,而抗氧化酶活性则显著升高;纳米颗粒在培养液中易发生聚集作用而沉积,12h后基本上完全沉积到底部。结果说明,50mg·L-1的纳米ZnO对浮萍产生了显著的胁迫作用,其对浮萍的毒性作用主要来源于其溶出的Zn2＋。     

基于K_means硬聚类算法的葡萄病害彩色图像分割方法

为了提高植物病害图像的分割精度与效果,根据植物病害症状及图像的特点,提出了一种基于K_means硬聚类算法（HCM）的葡萄病害彩色图像非监督性分割处理方法。该方法是在L*a*b*颜色空间模式下利用ab二维数据空间的颜色差异,以平方欧式距离作为像素间的相似度距离、以均方差作为聚类准则函数对颜色进行二分类聚类,并通过数学形态学运算对聚类结果进行校正。利用该方法对3种葡萄病害彩色图像进行分割的结果表明,该方法能够较为准确地将病斑区域从彩色图像中分割出来,对葡萄病害彩色图像的分割处理比较理想,鲁棒性好,分割准确率     

一品红不同品种叶片叶绿素荧光特性比较

 利用叶绿素荧光技术测定了一品红4个品种‘金多利’、‘圣诞天使’、‘索诺拉飞雪’、‘旗帜’叶片光强依赖的叶绿素荧光特性。结果表明: ‘金多利’初始荧光( F_o ) 、最大荧光( F_m ) 和可变荧光( F_v ) 值极显著高于其它3个品种。在各个光强( PAR, 0～2 171μmol·m^{- 2} ·s^{- 1} ) 下, 表观电子传递速率( rETR) 、实际光化学效率(Yield) 和光化学淬灭( q_P ) 的变化趋势一致, 均为‘金多利’ > ‘圣诞天使’ > ‘索诺拉飞雪’ > ‘旗帜’, 在高PAR下这种差异更明显。拟合参数最大潜在相对电子传递速率( rETR_max ) 、半饱和光强( Ik) 和光抑制参数(β) 也同rETR等具有一致变化。说明‘金多利’有较高的PSⅡ活性, 耐强光; 而‘旗帜’则光化学效率较弱, 高光强下易受光抑制。     

Sinomenine reduces MPP+-induced damage in SK-N-SH cells via ANRIL/miR-626 signaling pathway

AIM To investigate the effect of sinomenine (SN) on the damage of human neuroblastoma SK-N-SH cells induced by 1-methyl-4-4 phenylpyridine (MPP⁺) and its mechanism for exploring the pathogenesis of Parkinson disease. METHODS SN was used to treat MPP⁺-induced SK-N-SH cells. The levels of malondialdehyde (MDA) and glutathione (GSH) in cell culture supernatants were measured by ELISA. The apoptosis was analyzed by flow cytometry. The protein expression levels of Bcl-2 and Bax were determined by Western blot. The expression levels of long noncoding RNA ANRIL and microRNA-626 (miR-626) were detected by RT-qPCR. Dual-luciferase reporter assay was used to evaluate the relationship between ANRIL and miR-626. After ANRIL small interfering RNA was transfected into SK-N-SH cells, the effects of ANRIL expression knock-down on MPP⁺-induced SK-N-SH cell apoptosis, the protein expression levels of Bcl-2 and Bax, and the levels of MDA and GSH in cell culture supernatants were examined. RESULTS After treatment with MPP⁺, the apoptotic rate, Bax protein level and ANRIL expression in SK-N-SH cells were increased (P<0.05), and the Bcl-2 protein level and miR-626 expression were decreased (P<0.05). The level of MDA in cell culture supernatants was increased (P<0.05), and the level of GSH was decreased (P<0.05). After SN treatment or ANRIL expression knock-down, decreased apoptotic rate, Bax protein level and ANRIL expression (P<0.05), and increased Bcl-2 protein level and miR-626 expression in MPP⁺-induced SK-N-SH cells were observed (P<0.05). The level of MDA in the cell culture supernatants was decreased (P<0.05), and the level of GSH was increased (P<0.05). CONCLUSION SN attenuates MPP⁺-induced damage in SK-N-SH cells by regulating ANRIL/miR-626 signaling pathway.