Effects of hysteresis on redistribution of soil moisture and deep percolation at continuous and pulse drip irrigation

The infiltration and redistribution of soil moisture under surface drip irrigation considering hysteresis were investigated in two soils (loamy sand and silt loam) of different texture. The effect of continuous versus intermittent application of 1, 2 and 4 l/h to the soils was evaluated in terms of wetting front advance patterns and deep percolation under the root zone. For this purpose, a cylindrical flow model incorporating hysteresis in the soil water retention characteristic curve, evaporation from the soil surface, and water extraction by roots was used. The results show that, compared with continuous irrigation, pulse irrigation slightly reduces the water losses under the root zone in both cases (with and without hysteresis). Also, at the total simulation time, in both types of irrigation, hysteresis reduces significantly the water losses under the root zone. Finally, the effect of hysteresis was found to be greater at higher discharge rate (4 l/h) and consequently at higher water content at the soil surface.     

锦鲤保护性组织RNA提取及引物扩增研究

从养殖锦鲤各种组织中提取完整的RNA, 是分子生物学研究锦鲤应激表达、体色变异、生长发育等功能基因表达的关键实验。在不伤害养殖锦鲤个体健康的情况下, 取用鱼体的鳍条、鳞片、鳃丝、血液4种保护性组织,可以最大程度地减少对珍贵锦鲤个体的损伤, 并达到研究相关基因表达的目的。本研究利用TR izo l法提取RNA, 获得的各样品RNA条带完整、纯度高。结果表明, 4 种保护性组织提取的RNA 经过逆转录反应, 得到了高质量的CDNA。内标基因( B- actin)、热激蛋白基因( hsp70)、金属硫蛋白基因(MT)扩增得到清晰的条带, 通过NCB I数据上的BLAST比对证明, 各序列的同源性在95%以上。利用本研究的方法, 可以充分保护养殖锦鲤个体的完好性, 用于进一步的分子生物学研究。     

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.     

Progress in roles of MNX1 antisense RNA 1 in malignant tumor

MNX1 antisense RNA 1 （MNX1-AS1） is a newly discovered long non-coding RNA （lncRNA）, which is highly dysregulated in various carcinomas and its expression level is closely related to the overall survival and prognosis of patients. MNX1-AS1 regulates the occurrence and development of carcinomas by endogenous competitive adsorption of miRNA, regulating cell cycle, inducing epithelial mesenchymal transformation and activating multiple signaling pathways. The in-depth study of the carcinogenesis of MNX1-AS1 is useful for the early diagnosis, targeted therapy and prognostic assessment of relevant carcinomas. This article reviews the roles of MNX1-AS1 in malignant tumor.     

Effect of lncRNA FEZF1-AS1 on viability and apoptosis of LPS-induced vascular endothelial cells by regulating miR-363-3p expression

AIM To investigate the mechanism of long noncoding RNA （lncRNA） FEZF1-AS1 regulating microRNA-363-3p （miR-363-3p） on the viability and apoptosis of lipopolysaocharide （LPS）-induced vascular endothelial cells. METHODS Human umbilical vein endothelial cells （HUVECs） were cultured in vitro. pcDNA-NC, pcDNA-FEZF1-AS1, anti-miR-NC, anti-miR-363-3p, miR-NC and miR-363-3p mimics were transfected into the HUVECs and LPS stimulation was applied for 24 h. RT-qPCR was used to detect the expression of FEZF1-AS1 and miR-363-3p. The cell viability was measured by MTT assay. The apoptotic rate was analyzed by flow cytometry. The dual-luciferase reporter experiment was used to verify the targeted regulation of FEZF1-AS1 and miR-363-3p. Western blot was used to determined the expression of cyclin D1, Ki67 and cleaved caspase-3. RESULTS Compared with control group, the expression level of FEZF1-AS1 in LPS group was significantly reduced （P<0.05）, and the expression level of miR-363-3p was significantly increased （P<0.05）. Compared with pcDNA-NC+LPS group, the cell viability in pcDNA-FEZF1-AS1+LPS group was significantly increased （P<0.05）, the apoptotic rate was significantly reduced （P<0.05）, the protein levels of cyclin D1 and Ki67 were significantly increased （P<0.05）, and the protein level of cleaved caspase-3 was significantly reduced （P<0.05）. Compared with anti-miR-NC+LPS group, the cell viability in anti-miR-363-3p+LPS group was significantly increased （P<0.05）, the apoptotic rate was significantly reduced （P<0.05）, the protein levels of cyclin D1 and Ki67 were significantly increased （P<0.05）, and the protein level of cleaved caspase-3 was significantly reduced （P<0.05）. Dual-luciferase reporter experiment confirmed that FEZF1-AS1 targeted miR-363-3p. Compared with miR-NC+pcDNA-FEZF1-AS1+LPS group, the cell viability in miR-363-3p+pcDNA-FEZF1-AS1+LPS group was significantly reduced （P<0.05）, the apoptotic rate was significantly increased （P<0.05）, the protein levels of cyclin D1 and Ki67 were significantly reduced （P<0.05）, and the protein level of cleaved caspase-3 was significantly increased （P<0.05）. CONCLUSION Over-expression of FEZF1-AS1 promotes the viability and inhibits apoptosis of LPS induced vascular endothelial cells by inhibiting the expression of miR-363-3p.     

大豆质核互作雄性不育系NJCMS1A与其保持系NJCMS1B的生理生化特性比较分析

以大豆质核互作雄性不育系NJCMS1A及其保持系NJCMS1B的花芽为试验材料,利用试剂盒法和紫外分光光度计测定分析了糖分含量(可溶性糖和淀粉)和总ATPase、蔗糖磷酸合成酶(SPS)、磷酸烯醇式丙酮酸羧激酶(PEPCK)、抗坏血酸氧化酶(AO)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)以及过氧化物酶(POD)等酶活性的变化情况。结果表明:相较于保持系NJCMS1B,总ATPase、PEPCK、AO和CAT活性在不育系NJCMS1A中显著下降;糖分含量、SPS和SOD活性在不育系NJCMS1A中下降水平不显著;POD活性在不育系NJCMS1A中显著上升。根据结果分析推测,与能量代谢或胁迫响应等相关的物质或酶活性在不育系NJCMS1A中的亏损现象可能是NJCMS1A花粉败育的主要原因。     

水稻齿叶矮缩病毒Pns10蛋白在水稻原生质体内的表达

【目的】水稻齿叶矮缩病毒(Rice ragged stunt virus,RRSV)Pns10蛋白在介体昆虫细胞内可形成类似病毒原质(viroplasm)的内含体,是RRSV侵染介体所必需。然而Pns10蛋白在水稻寄主中是否具有类似功能及其表达情况如何未见报道。【方法】利用大肠杆菌系统表达Pns10蛋白,免疫家兔制备多克隆抗体;通过水稻原生质体病毒侵染体系,利用免疫荧光技术分析Pns10蛋白在水稻原生质体内的分布情况,利用实时定量PCR技术和Western blot技术分别检测Pns10 RNA和Pns10蛋白在水稻原生质体内的积累情况。【结果】将Pns10基因克隆到Gateway系统原核表达载体p DEST17中,IPTG诱导表达成功后,制备融合蛋白抗血清。Western blot检测显示,该抗血清可检测感病水稻叶片中的Pns10蛋白。病毒侵染水稻原生质体后,Pns10蛋白可形成类似病毒原质的内含体;Pns10 RNA在病毒接种8 h后开始积累,24 h后达到最大值,随后开始下降;Pns10蛋白在24 h后开始表达,之后维持较高水平,60 h后略有下降。【结论】成功获得了Pns10抗血清;Pns10在水稻原生质体内成功表达,可形成类似病毒原质的内含体,并且Pns10 RNA的表达先于其蛋白的表达。     

寄主诱导的基因沉默提高植物真菌病害抗性研究进展

寄主诱导的基因沉默(host-induced gene silencing,HIGS)以病原菌生长发育、产孢繁殖、侵染或致病过程中的关键基因作为靶点,在寄主植物中表达针对靶基因的RNAi构建体,在病原菌侵染植物的过程中,摄入相应的ds RNA或si RNA分子,通过识别、结合特异核苷酸序列,干扰病菌靶基因表达,从而抑制病菌侵染和扩展,使植物表现抗病。这项技术为培育基于病原菌特异序列的植物抗病性奠定了基础,显示了巨大的应用潜力。本文综述了利用HIGS技术提高植物真菌病抗性的最新研究进展,总结了国内外利用这项技术改良植物真菌病害抗性的主要策略、技术路线,展望了发展应用前景。     

Effects of CENP-W down-regulation on human glioma U87 cells

AIM: To study the effect of centromere protein W (CENP-W) down-regulation on human glioma U87 cells.METHODS: Small interfering RNA (siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot. The proliferation of the cells was analyzed by MTT assay, BrdU staining and colony formation experiment. Transwell chamber assay was used to detect the invasion ability of the cells. The cell migration ability was measured by a scratch test. The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry.RESULTS: The results of MTT assay, colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group. The results of Transwell and scratch experiments showed that the migration and invasion abilities in experimental group were weaker than those in blank control group and negative control group. The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G₀/G₁ phase. The percentage of apoptotic cells in experimental group was higher than that in control group (P<0.05).CONCLUSION: Down-regulation of CENP-W expression inhibits the proliferation, migration and invasion of human glioma cells and promotes the apoptosis of the cells, suggesting that CENP-W may be a potential target of gene therapy for human glioma.