菜薹BrNAP克隆及其在采后叶片衰老中的功能分析

在菜薹(Brassica rapa var. parachinensis)中克隆Br NAP1并分析其功能。Br NAP1编码区全长813 bp,编码270个氨基酸,具有NAP转录因子特有的保守结构域,属于NAP亚家族成员。Br NAP1表达量与叶片衰老程度呈正相关且受ABA诱导表达上调。亚细胞定位试验表明Br NAP1定位于细胞核。互补试验显示Br NAP1能使拟南芥atnap滞绿表型回复至野生型,过表达则能引起采后叶片早衰。双荧光素酶试验表明Br NAP1能够激活Br SAG113表达。这些说明Br NAP1是菜薹采后叶片衰老的正调控基因。     

Caveolae-mediated endocytosis of VE-Cad contributes to vascular hyperpermeability after LPS treatment

AIM: To observe caveolae-mediated endocytosis of vascular endothelial cadherin (VE-Cad) after lipopolysaccharide (LPS) treatment, and its role in vascular hyperpermeability. METHODS: Human vascular endothelial cell line CRL-2922 was used in the experiment. Western blot, co-immunoprecipitation and immunocytochemistry were adopted to observe the protein expression of caveolin-1 (Cav1), a main structural protein of caveolae, after LPS treatment. Caveolae-mediated endocytosis of VE-Cad after LPS treatment, and the effects of caveolae inhibitor on caveolae-mediated endocytosis of VE-Cad, the protein expression of VE-Cad and monolayer cell permeability after LPS treatment were determined. RESULTS: After LPS treatment, the protein expression of Cav1 did not show a significant change, while the phosphorylation (Tyr14) of Cav1 was significantly increased (P<0.05). No co-immunoprecipitation or co-localization of VE-Cad with Cav1 was observed in the normal control, and that was increased time-dependently after LPS treatment (P<0.05). Filipin, an inhibitor of caveolae, at concentration of 5 mg/L significantly reduced the co-immunoprecipitation of VE-Cad with Cav1 (P<0.05), increased the expression of VE-Cad (P<0.05) in the membrane, and improved the monolayer cell permeability at 4 h after LPS treatment (P<0.05). CONCLUSION: Caveolae-mediated endocytosis of VE-Cad contributes to the internalization of VE-Cad and the monolayer cell hyperpermeability after LPS treatment.     

Time Effect Experimental Analysis of the Soil Arch Effect of Anti sliding Pile

Soil arching effect is important for stabilizing the soil behind anti sliding pile. The creep of soil mass will inevitably lead to the formation of soil arching, which has time effect. In this analysis, an indoor thrust pile model test is proposed. The characteristics of stress distribution in the soil and the time effect of soil arching behind anti sliding pile are analyzed. The experimental data acquired by the soil pressure cells arranged along the normal direction of the thrust show that soil arching effect is enhanced with the increase of thrust.The thickness of soil arch increases with time.And the data obtained by soil pressure cells arranged along the direction of the thrust show that the horizontal soil arching effect increases and then gradually extends.The scope of stress dispersion has very large growth along the normal direction as time goes on.     

Performance analysis of StimGun technology

With further exploration and development of oil and gas wells, it is found that the unconventional oil-gas bed occupies a major proportion in gas reserves. However, it is difficult to obtain satisfactory control by using conventional perforation technology because of its characteristics of low pore, low permeability and compact lithology in the unconventional oil-gas bed. The StimGun tech (synergize compound perforation) is described as following: after the perforating gun is fired, propellant burns with high-energy gas produced. Then the high-energy gas enters the perforating hole and forms multi-path cracks around perforated hole, which connects with the natural cracks and improves the permeability of oil and gas bed. The application and advantage of StimGun are introduced. The Stimgun is successfully applied in Sichuan LG oilfield. It can control the fracture heights effectively and remove contamination formation with no water producing, and it is also a great help to increase productivity of the oil-gas bed with low permeability.     

越橘葡萄座腔菌枝枯病的病原菌鉴定

 通过形态学观察、诱导产孢、致病力测定和分子生物学研究,对引起辽宁地区越橘枝枯病 的病原菌进行鉴定,确定其为葡萄座腔菌（Botryosphaeria dothidea）。病原菌在越橘植株上接种后表现枝 条枯萎、木质部变色等症状,与田间自然发病植株一致;在PDA 培养基上菌落初期为白色,后转为灰黑 色,分生孢子纺锤形或梭形,无色,单胞,内含多个油滴;水琼脂 + 麦芽提取物、苹果枝条和苹果幼果 3 种病原菌培养方法均可诱导产孢,苹果幼果是诱导病原菌产孢的最佳基质。供试菌株LB1 的rDNA-ITS 和β-tubulin 基因序列与GenBank 中已有的Botryosphaeria dothidea 序列相似性分别达到99%与100%。     

Effects of FKBP12.6 gene on canine heart failure transfected by ultrasound mediated destruction of microbubbles

AIM: To investigate the effect of gene transfection of FKBP12.6 (FK12.6 binding protein) on the canine failing heart induced by rapid ventricular pacing. METHODS: Three weeks after onset of rapid ventricular pacing (250 bpm), 28 canines were divided into 4 groups. Either pcDNA3.1-FKBP12.6 plasmid encoding human FKBP12.6 gene (transfection groupⅠ,Ⅱ; observed at the 4th or 14th days respectively after transfection) or empty vector (controlⅠ,Ⅱ) was transferred into myocardium under ultrasonic microbubble destruction. After transfection, maintenance pacing at a reduced rate (190 beats/min) was continued until end-point of experiment. Echocadiographic, hemodynamic and plasmic ANP (atrial natriuretic peptide), BNP (brain natriuretic peptide) data were collected three times (before pacing, before transfection and endpoint). Semiquantative RT-PCR was used to identify FKBP12.6 expression in myocardium. RESULTS: Gene transfection of FKBP12.6 elevated expression of FKBP12.6 mRNA 3.5 folds at day 4, and 1.7 folds at day 14 respectively, compared with control group. Significant improvements of cardiac function, hemodynamics and plasmic concentrations of ANP, BNP were observed in transfection groupⅠ and these effects were stable for at least 2 weeks. Cardiac output (CO), ejection fraction (EF) and fractional shortening (FS) were still lower than pre-pacing although they increased in transfection groups. Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-diastolic volume (LVEDV) decreased at day 14 but LVEDD remained unchanged at day 4 compared with pre-transfection. CONCLUSION: Gene transfection of FKBP12.6 improves cardiac functions in the failing heart and reverses myocyte remodeling. This might be a novel therapeutic application for treating human heart failure.     

Role of Rho kinase in the dysfunction of myocardium contraction in hemorrhagic shock rats

AIM: To explore the role of Rho kinase in myocardium contraction in hemorrhagic shock rats. METHODS: The rat cardiac papillary muscle and rat heart at different time points during hemorrhagic shock were isolated. The contraction of papillary muscle in response to isoprenaline and the hemodynamic parameters of the isolated hearts in vitro including left intraventricular systolic pressure (LVSP), the maximal change rate of left intraventricular pressure (±dp/dtmax) were measured. RESULTS: The activity of Rho kinase, the contractile response of cardiac papillary muscle to isoprenaline and the hemodynamic parameters of the hearts were decreased gradually at the 1 h, 2 h and 4 h after shock. Compared to the normal controls, the activity of Rho kinase, the contractile response of cardiac papillary muscle and the hemodynamic parameters of the hearts were decreased significantly. The change of Rho kinase was positively correlated with the contractile response and the hemodynamic parameters of the heart. Stimulation with U-46619 at concentration of 10-8 mmol/L increased the activity of Rho kinase and improved the contractile response of cardiac papillary muscle to isoprenaline and the hemodynamic parameters of the isolated heart. The effects of U-46619 on the above parameters were abolished by Rho kinase specific inhibitor Y-27632 at concentration of 10-5 mol/L. CONCLUSION: Rho kinase plays an important role in the regulation of cardiac contractility during hemorrhagic shock.     

重庆宜居城市建设中的法律保障体系探析

宜居城市的建设是一个包括自然、经济、社会在内的、复杂的综合系统工程。城市的资源和环境容量是有限的,宜居城市的建设和发展必然会受到环境和资源条件的制约,为了实现可持续发展、建设社会主义新型和谐社会,在宜居城市的建设中必须要考虑环境和资源的承载力,而法律的引导、规划、强制和保障必不可少。文中拟以重庆市为例,从环境保护和城市规划两个方面着手,分析重庆宜居城市建设中的法律保障体系的脉络并提出一定的建议,希望能够对重庆宜居城市的建设有所裨益。     

丘陵山区基本农田空间布局优化及评价

划定基本农田、实行永久保护已上升为国家战略;但基本农田划定和质量评定工作依然面临许多技术挑战。以重庆荣昌县为研究区,在地理信息系统技术支持下,通过构建耕地质量综合评价模型和引入景观格局分析方法,探讨了丘陵山区开展基本农田划定和质量评定工作的技术方法与手段。研究结果表明:①丘陵山区基本农田布局调整更具综合性,不仅要结合土地资源条件,还要考虑经济社会发展因素和生态建设需求;②通过基本农田空间布局调整和综合质量评价,荣昌县划定基本农田面积48 717.40hm2,基本农田质量分值较调整前增加19.73;③从景观格局来看,调整后基本农田的斑块破碎度降低、连片性提高,为发展现代农业和规模经营创造了条件。研究认为,依托地理信息系统技术平台,综合耕地质量评价和景观格局分析方法,能够为丘陵山区基本农田划定和质量评定工作提供科学支撑。