Effect of inhibiting nuclear factor-kappa B on TNF-α-induced expression of angiotensinogen and production of angiotensinⅡ in glomerular mesangial cells

AIM: To study the effect of inhibiting nuclear factor-kappa B (NF-κB) activity on the expression of angiotensinogen (AGT) and the production of angiotensinⅡ (AngⅡ) induced by tumor necrosis factor-α (TNF-α) in glomerular mesangial cells (MCs) of SD rats. METHODS: The MCs of SD rats were isolated and divided into three groups as follows: control; MCs treated with TNF-α, and the MCs treated with TNF-α + pyrrolidinedithiocarbamate (PDTC). The activity of nuclear factor-kappa B was measured by electrophoretic mobility shift assay. The expression of AGT was determined by RT-PCR for mRNA and Western blotting for protein. The concentration of angiotensinⅡ in supernatant was measured by RIA. RESULTS: The NF-κB activity in the MCs treated with TNF-α (20.67±9.14)×102 μg/cell was significantly higher than that in control cells ［(8.25±4.35)×102 μg/cell, P<0.01］ and the MCs treated with TNF-α+PDTC ［(7.20±4.57)×102 μg/cell, P<0.01］, and no significant difference was found between control and the MCs treated with TNF-α＋PDTC (P>0.05). The AGT mRNA level in the MCs treated with TNF-α (0.27±0.05) was higher than that in the control cells (0.20±0.05, P<0.05), and no significant difference was observed when compared with that in the MCs treated with TNF-α＋PDTC (0.22±0.06, P>0.05). The expression of AGT protein in the MCs treated with TNF-α (0.60±0.19) μg/cell was higher than that in the control ［(0.37±0.15)μg/cell, P＜0.05］ and the MCs treated with TNF-α＋PDTC ［(0.37±0.17)μg/cell, P<0.05］, and no significance was found between the MCs treated TNF-α＋PDTC and the control (P>0.05). The AngⅡ level in supernatant of cultured MCs treated with TNF-α ［(9.73±2.38)×10-5 ng·L-1/cell］ was significantly higher than that in the control ［(7.50±1.51)×10-5 ng·L-1/cell, P<0.05］ and in the MCs treated with TNF-α＋PDTC ［(6.94±1.46)×10-5 ng·L-1/cell, P<0.05］, however, the difference between the MCs treated with TNF-α＋PDTC and the control was of no significance (P>0.05). CONCLUSION: TNF-α activates the NF-κB in glomerular MCs, induces the AGT expression and the production of AngⅡ. Inhibition of NF-κB decreases the AGT expression and the production of AngⅡ. Therefore, the effects of TNF-α on AGT and AngⅡ may be mediated by NF-κB.     

Effects of cytokines on renovation of acute renal failure in mouse with bone marrow derived mesenchymal stem cell transplantation

AIM: To observe the effects of cytokines on renovation of acute renal failure (ARF) in mouse with bone marrow derived mesenchymal stem cell (MSC) transplantation. METHODS: ARF animal model was induced in mouse by subcutaneous injection of cisplatin. Mice were randomly assigned into 3 groups: normal control group, ARF group and MSC group. After 24 h cisplatin injection, animals were injected intravenously with MSC in MSC group. Animals were sacrificed at 1 d, 4 d, 7 d, 14 d and 28 d after cisplatin injection. The blood urea nitrogen (BUN) and serum creatinine (Scr) were measured. The renal morphologic changes were scored with Paller’s criterion on hematoxylin and eosin (HE) stained sections. The mRNA and protein expressions of HGF, BMP-7, TNF-α and IL-10 were detected by RT-PCR and immunohistochemistry method. RESULTS: After 4 d of cisplatin injection, the BUN and Scr values in MSC group were significantly lower than those in ARF group (P<0.01). After 7 d and 14 d, the values of BUN and Scr in MSC group were still lower than those in ARF group (P<0.01, P<0.05). The renal morphologic scores of MSC group were also lower than those of ARF group. After 7 d, the expressions of HGF, BMP-7 and IL-10 were higher in MSC group than those in ARF group, the expression of TNF-α in MSC group was lower than that in ARF group. CONCLUSION: MSC promotes the recovery of acute renal failure induced by cisplatin. The mechanism may partly depend on paracrine of growth factor and amelioration of inflammatory.     

干花染色技术研究

研究了云南省常见的几种漂白过的花材的染色工艺.结果表明:染液的浓度增加,染色效果显著,染液浓度为3‰时可获得满意效果.温度升高及时间增加,有利于花材的染色,当温度达到70～90℃时染色效果最佳,染液pH在4～6之间为佳.     

香菇中γ-谷氨酰转肽酶的分离纯化及其酶学性质研究

通过(NH4)2SO4沉淀、Phenyl Sepharose FF疏水层析分离纯化香菇中的γ-谷氨酰转肽酶(γ-Glutamyltranspeptidase,GGT),纯化后的GGT的比活力到达了19.59 U/mg.SDS-PAGE分析表明,GGT由分子量分别为28 kDa和60 kDa的两个亚基组成.酶学性质试验结果表明,GGT反应的最适温度为37 ℃,最适pH 值为7.6;金属离子Na+、K+和Ca2+对酶有激活作用,而Ag+、Cu2+、Zn2+和Fe3+则有抑制作用;试验范围内,GGT水解γ-glutamyl-p-nitroanilide的米氏动力学参数Km 值为2.601 μmol/mL,Vmax值为0.0287 μmol/min;组成的氨基酸中,Glu和Asp含量较高,Met和Cys含量较低.     

无公害露地蔬菜诊断施肥技术研究

本试验选择我区大面积露地种植的四大蔬菜品种,进行不同氮、磷、钾肥施用量及施用时期的实验,通过室内肥残检测,探讨氮、磷、钾肥在我区无公害蔬菜生产上合理用量和使用时期,实现降低成本和肥残,提高产量,达到食品安全的目的。     

新型挂面包装机的输送和称重设计

在许多领域的应用中,自动称重扮演一个重要组成部分,在交易中心大量的商品和原料的处理,散装到散装称重过程使用自动皮带秤或总计料斗秤,数量较小的商品则通过重力灌装机械或分检秤自动称重和灌装到终端用户．由车辆运送或经铁路运输的商品往往分别用运动道路称重装置或自动轨地秤称重。一种新型挂面包装机采用带式输送装置、称重传感器,使挂面包装机结构简洁,输送稳定,称重精确,易于实现自动化控制,具有良好的性价比。     

二氧化硅改性处理桉树生材的材性分析

采用溶胶-凝胶法,对含水率142％～146％的速生桉树生材进行改性处理,采用电镜SEM、X-射线能谱仪DAX及红外光谱FTIR分析,观察木材微观结构和SiQ分布状态,并检测理化性能.结果表明:浸渍时间不同,改性材中的SiO2含量和分布不同;改性材的增重率最大达45％,气干密度可提高至0.6 g/cm3左右,抗弯强度、硬度及耐光老化性能等均相应提高.     

茶毛虫核型多角体病毒不同分离株的毒力水平与遗传结构关系分析

4个茶毛虫核型多角体病毒（EpNPV）分离株对茶毛虫的毒力测定结果表明,浙江分离株的毒力最强,其LC₅₀为2.5×10⁶PIB/mL;湖南分离株的毒力最弱,其LC₅₀为13.36×10⁶PIB/mL。不同分离株的毒力强弱依次为浙江分离株>贵州分离株>湖北分离株>湖南分离株,但贵州分离株与湖北分离株的毒力相差不大,LC₅₀分别为9.5×10⁶PIB/mL和7.31×10⁶PIB/mL。测定了EpNPV 4个分离株的5个与病毒复制、病毒—宿主关系、毒力水平相关的功能基因序列,分析其不同株系间的遗传结构,发现湖北分离株和贵州、湖南两分离株的相似度最高（99.7%）;浙江分离株和湖南分离株间的相似度最低（99.4%）。基于5个基因拼接序列构建的系统进化树显示,湖南（HN）和湖北（HB）两个株系最先聚类并为一支,再与贵州（GZ）分离株聚类,最后才与浙江分离株（ZJ）聚合,说明浙江分离株与其他3个分离株的遗传距离均较远。试验结果初步表明,EpNPV不同株系间的毒力水平差异与其遗传结构差异明显相关。     

水稻条纹叶枯病的为害损失及防治指标

2004-2006年在浙江北部系统地调查了水稻条纹叶枯病发病动态和为害损失。单季晚稻秧田期介体灰飞虱有效虫量（X2）与水稻株发病率（Y1）的关系式为Y1=0.0873 + 1.0757X2,晚稻本田株发病率（X）与产量损失率（Y）总关系式为Y=0.1254+0.7551X。在现有生产条件下,经济允许水平以损失率表示为2.0%~2.5%,制订了水稻条纹叶枯病防治指标为水稻秧苗期和本田前期介体灰飞虱有效虫量2~3头/m2,该指标已在生产上大面积推广应用。     

一个小麦AP2/ERF转录因子家族单独亚族基因的克隆及分析

AP2/ERF家族转录因子是植物中重要的一类转录因子,广泛参与植物各类生理过程.为了给该家族转录因子的深入研究提供基础,利用小麦EST数据库(http://www.ncbi.nlm.nih.gov/UniGene/UGOrg.cgi?TAXID= 4565),以拟南芥AP2/ERF家族转录因子单独亚族成员AtAP2-Soloist的氨基酸序列为探针,搜索到一个小麦UniGene数据,经拼接得到1个全长cDNA序列(TaAP2-Solosit),通过RT-PCR方法从"扬麦15"的cDNA中克隆了该转录因子基因(YM15AP2-Solosit).克隆的转录因子核苷酸序列与电子克隆的序列只有1个碱基不同,两者编码氨基酸序列一致.从cDNA序列、氨基酸序列的相似性、组成成分、理化性质、疏水性/亲水性、进化树、功能域、二级和三级结构等方面进行了分析,结果显示"扬麦15"中的YM15AP2-Solosit转录因子属于AP2/ERF家族中的单独亚族,是亲水性蛋白,在蛋白质的三级结构上与拟南芥的AtAP2-Soloist相似.EST丰度分析显示,YM15AP2-Solosit在根、茎、叶和花中均有表达.