Effect of miR-19a on lipid catabolism in hepatocyte LO2

AIM: To observe the effect of microRNA-19a (miR-19a) on the lipid catabolism of hepatocyte LO2, and to explore the potential mechanism. METHODS: miR-19a was over-expressed or silenced by transfection of miR-19a mimics or miR-19a inhibitor into LO2 cells, then the mRNA level of miR-19a was detected by real-time PCR. The potential target of miR-19a was found by the method of bioinformatics through internet website. The effect of miR-19a on the 3' UTR of peroxisome proliferator-activated receptor α (PPARα) was measured by dual luciferase reporter assay, and the protein level of PPARα and its 2 major downstream rate-limiting enzymes involved in lipid catabolism, acyl-coenzyme a dehydrogenase (ACADM) and carnitine palmitoyltransferase 1A (CPT1A), were detected by Western blotting. Meanwhile, the effect of miR-19a on the generation of ketone body was measured by beta-hydroxybutyric acid (β-OHB) detection assay. RESULTS: The mRNA level of miR-19a was dramatically elevated by the transfection of miR-19a mimics, and sharply decreased by the transfection of miR-19a inhibitor (P<0.05). PPARα was found as a potential target of miR-19a, and dual luciferase reporter assay and Western blotting confirmed the regulatory effect of miR-19a on the expression of PPARα, with the protein level changes of ACADM and CPT1A. miR-19a mimics down-regulated, while miR-19a inhibitor up-regulated the concentration of β-OHB in LO2 cells (P<0.05). CONCLUSION: miR-19a regulates the lipid catabolism of hepatocytes by targeting the PPARα and its 2 downstream rate-limiting enzymes.     

Fusarium solani Upregulated Sesquiterpene Synthase Expression,Sesquiterpene Production and Allelopathic Activity in Piper betle L.

Activators of sesquiterpene synthase (STS) gene expression and sesquiterpene production in Piper betle L. were examined using quantitative real time PCR and gas chromatography mass spectrometry methods, and the allelopathic activity of untreated and Fusarium solani-treated betel extracts was tested on seed germination and on the shoot and root growth of Thai rice variety PSL2 (Oryza sativa cv. Phitsanulok 2) and three dominant paddy weeds (Eclipta prostrata, Echinochloa crus-galli and Chloris barbata). The results demonstrated that F. solani dramatically upregulated STS expression and productions of β-cubebene, β-caryophyllene and germacrene D sesquiterpene when compared with the untreated control, and that betel extracts had a greater inhibitory effect on weeds than on rice. The effects were more clearly detected on seed germination and root growth than on shoot growth, and they were found to be dose-dependent. It is also noted that F. solani-treated extract had stronger effects than the untreated extract. The species most sensitive to the allelopathic effects was C. barbata, germination of which was completely inhibited even at a dose of 0.1 mg/mL untreated extract. With regards to rice, although betel extract at 1.0 mg/mL showed no inhibition on germination, it affected the elongation of rice roots, in addition to those of the tested weeds. The obtained data suggested that F. solani has potential as an activator of sesquiterpene allelochemical production via STS expression, the latter leading to the treated betel extract having a stronger phytotoxic effect. These results were beneficial in the promotion of natural herbicide production using biotechnology.     

关于藏书倒架的研究与探讨——以中国矿业大学图书馆文昌科技图书阅览室为例

结合工作实际探讨藏书倒架的前期准备工作及倒架时的路线设计、人员组织安排和倒架后的环节完善。以期减轻图书馆人员在倒架工作中的劳动强度和工作量，提高工作效率。     

国内乡村生态旅游研究综述

乡村生态旅游作为推动乡村经济发展的新形式已经成为乡村发展研究的新热点。以近 20年内发表于SCI、CSSCI和核心期刊的 225篇文献为研究对象发现，国内乡村生态旅游相关研究划分为基础性研究和宏观性研究两大类。分别对其相关概念、理论基础、影响因素和发展模式做出基础性阐释，对其开发管理、可持续发展、利益相关者和相关评价体系进行理论性研究发现，现阶段的研究在研究领域、研究范围、研究方法方面得到发展，但在研究内容和案例选址方面仍存在缺陷，需平衡各个研究领域，不断完善相关研究，精细挑选案例地址，推动中国乡村生态旅游研究。     

Effect of the Substrates on the Production of Engrafted Vine Cuttings in Heated Greenhouses

This article highlights the results of a long-term research project on the production of grape stalks engrafted by the desk method. The integrated impact of rootstock varieties, modes of stratification, and different substrates on vegetating vine cuttings, raised in heated greenhouses, and the capacity of their acclimatization after planting was studied. The results of the research show that rootstock variety and modes of stratification influence the output of engrafted vegetating cuttings of vines grown in pots using various substrates. In particular, the engrafted cuttings of variety Rkatsiteli produced on two rootstocks, 5 BB and 101-14, passed stratification (a) in sawdust with local electroheating, (b) on the water with its periodic change, and (c) in the layer of perlite. After preplanting preparations they were planted in pots with six different substrates: (1) soil (control); (2) perlite; (3) sawdust; (4) rice husk + soil + sand (1:1:1); (5) peat + soil + sand (1:1:1); and (6) mold + soil + sand (1:1:1). The cuttings were grown with a covered root system in a heated greenhouse for 35–40 days. Forty-day-old vegetating cuttings, after hardening, were planted into the open ground. Specialties were established during the root and shoot formation on vegetating nursery plant grafts during the rooting period. Optimal substrates for growing engrafted cuttings with a covered root system in heated greenhouses for each rootstock and stratification mode were determined.     

教学督导听课技巧与评课策略分析——督导听课心得

通过3位优秀教师的讲课风格诠释了授课艺术,通过自己督导期间听课实践总结出听课技巧;吸纳前人经验,提出评课策略及应该注意的问题。     

microRNA-146a通过靶向MAPK4和Myosin Va基因调控羊驼黑色素细胞增殖、迁移

旨在研究microRNA-146a(miR-146a)对羊驼黑色素细胞增殖和迁移的调控及其分子机制。本研究使用双荧光素酶试验验证MAPK4和Myosin Va是miR-146a的靶基因;利用荧光定量PCR和蛋白质免疫印迹试验检测在羊驼黑色素细胞中过表达miR-146a对相关下游基因表达的影响;利用CCK8和Transwell检测miR-146a过表达对羊驼黑色素细胞增殖和迁移的影响。结果显示,与对照组相比,将miR-146a和MAPK4或Myosin Va共转染293T细胞,双荧光素酶活性分别极显著下降36%和30%(P<0.001);MAPK4和Myosin Va基因转录水平分别极显著下调67%和47%(P<0.001,P<0.01);蛋白质水平的表达量分别显著或极显著下调38%和69%(P<0.05,P<0.01);增殖和迁移相关的基因CREB、MITF、MLPH和Rab27a在转录水平和蛋白水平的表达均极显著下调(P<0.01,P<0.001);CCK8和Transwell结果显示,过表达miR-146a使羊驼黑色素细胞的增殖和迁移能力极显著下调(P<0.01)。综上所述,miR-146a通过靶向调控MAPK4和Myosin Va,使增殖和迁移相关的基因MEK1、CREB、MITF、MLPH和Rab27a的表达下调,从而对羊驼黑色素细胞的增殖和迁移起抑制作用。     

AR regulates porcine immature Sertoli cell growth via binding to RNF4 and miR-124a

Sertoli cells are the only somatic cells in the seminiferous epithelium which directly contact with germ cells. Sertoli cells exhibit polarized alignment at the basal membrane of seminiferous tubules to maintain the microenvironment for growth and development of germ cells, and therefore play a crucial role in spermatogenesis. Androgens exert their action through androgen receptor (AR) and AR signalling in the testis is essential for maintenance of spermatogonial numbers, blood–testis barrier integrity, completion of meiosis, adhesion of spermatids and spermiation. In the present study, we demonstrated that AR gene could promote the proliferation of immature porcine Sertoli cells (ST cells) and the cell cycle procession, and accelerate the transition from G1 phase into S phase in ST cells. Meanwhile, miR-124a could affect the proliferation and cell cycle procession of ST cells by targeting 3′-UTR of AR gene. Furthermore, AR bound to the RNF4 via AR DNA-binding domain (DBD) and we verified that RNF4 was necessary for AR to regulate the growth of ST cells. Above all, this study suggests that AR regulates ST cell growth via binding to RNF4 and miR-124a, which may help us to further understand the function of AR in spermatogenesis.