Gelatinases expression and their activity in rat alveolar macrophages induced by cigarette smoke medium

AIM: To study the effect of cigarette smoke medium (CSM) on the gene expression and activity of gelatinases from alveolar macrophages (AMs) in the rat, and then to explore their role in the pathogenesis of chronic obstructive pulmonary disease (COPD). METHODS: AMs were obtained from BALF of the rats that had smoked for 12 weeks. CSM was produced following the method of Wirtz and colleagues, and the cultured AMs were respectively stimulated for 24 h by 0%, 1%, 3%, 5%, 10%, 15% CSM. The mRNA levels of MMP-9 and MMP-2 were detected by semi-quantitative RT-PCR, and the enzyme activity was measured by Zymography. RESULTS: When the concentration of CSM was below 5%, the expression and activity of MMP-9 and MMP-2 signficantly increased with the concentration of CSM in a dose-depended manner (P<0.05). While the concentration of CSM exceeded 5%, the expression and activity of MMP-9 and MMP-2 correspondingly decreased with the increase in CSM concentrations (P<0.05), which possibly were related with the cytotoxicity of CSM. CONCLUSION: The expression and activity of MMP-9 and MMP-2 are induced by CSM. The gelatinases induced by smoking from AM may play an important role in the pathogenesis of COPD.     

Vaspin and insulin signaling pathways in obstructive sleep apnea-hypopnea syndrome animal model

AIM: To establish a suitable chronic intermittent hypoxia (CIH) model in rats for modeling the obstructive sleep apnea-hypopnea syndrome (OSAHS) and further to explore the relationship between vaspin and insulin signaling pathways at mRNA and protein levels. METHODS: Healthy male Wistar rats were divided into control group and CIH group. The rats in control group were raised under physiological condition, and the rats in CIH group were kept in the plexiglass chamber between 9:00~17:00 and underwent intermittent hypoxic challenge 8 h/d for 8 weeks. The blood pressure of arteria caudilis in the rats was measured by tail cannulation before and after study. Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), fasting insulin (FINS), vaspin and adiponectin levels were measured. The mRNA expression of vaspin was detected by real-time PCR. The protein levels of vaspin, Akt and p-Akt were determined by Western blotting.RESULTS: Before the experiment, the weight and blood pressure of the rats had no significant difference, while after the experiment the blood pressure in CIH group was obviously higher than that in control group. FPG, FINS, TG and TC in CIH group were also markedly higher than those in control group (P<0.05). Serum vaspin concentration was significantly correlated with FPG, FINS, HOMA-IR and TC. The expression of vaspin at mRNA and protein levels in the fat tissue of CIH group was evidently higher than that in control group. The protein levels of p-Akt decreased distinctly in CIH group. CONCLUSION: The levels of FINS, HOMA-IR, SaO2 and vaspin were markedly higher than those in control group, indicating that there was a close relationship between vaspin and insulin resistance in OSAHS.     

‘短柄樱桃’花芽休眠解除过程中差异表达基因的研究

 以中国樱桃的优良品种‘短柄樱桃’（Prunus pseudocerasus Lindl.‘Duanbing’）的花芽为材料,采用mRNA 差异显示技术,分析休眠解除的过程中相关差异表达基因。共克隆获得79 个差异cDNA片段;通过半定量RT-PCR 验证,确定18 个阳性差异cDNA 片段为休眠解除相关候选基因;休眠解除过程中上调的基因片段11 个,下调的7 个。测序及同源性比对结果显示：差异表达基因主要参与糖代谢、信号转导、跨膜转运及氧化还原等反应过程。这些基因在‘短柄樱桃’休眠解除阶段可能起到直接或间接的作用。     

气相色谱-嗅觉测量法鉴定蒌蒿中的香气化合物

 采用顶空固相微萃取法提取了蒌蒿样品中的挥发性成分, 利用气相色谱-嗅觉测量法(GC-O)并结合气相色谱质谱联用分析(GC - MS) , 鉴定了萎蒿中的香气化合物。共检出21个呈香化合物, 基于其嗅感强度, 反式- 2 - 己烯醛、水芹烯、顺式- 罗勒烯、3 - 崖柏酮、樟脑, 黄瓜醛、( - ) - 龙脑和( - ) - 乙酸龙脑酯被认定为形成蒌蒿香气的重要化合物。     

无锡地区鸡群禽流感疫苗免疫效果

对江苏省无锡地区鸡群使用禽流感H5和H9灭活疫苗或活疫苗后的免疫水平进行了监测。结果显示：母源抗体水平越低,抗体上升幅度越大,且抗体达到最高时所需的时间越短;H5单价灭活苗与H5＋H9二联灭活苗产生H5的抗体效果差异不大。通过对其他禽类和鸟类的临床样品检测发现：应加大对野禽及鸟类的监测力度,防止携带该病毒传播给家禽。这一监测结果对有效防控该病具有重要的实践价值。     

陆地棉一个丝氨酸/苏氨酸激酶蛋白基因的克隆与表达分析

通过RT-PCR获得一个编码棉花丝氨酸/苏氨酸激酶类似蛋白全长的cDNA片段,其编码的氨基酸序列与拟南芥ATPK3的相似度达到82%,这个基因暂被命名为GhPK1。基因全编码区包括1038个核苷酸,编码346个氨基酸的蛋白。GhPK1的编码产物在265~270个氨基酸处有一个跨膜区并可能结合在内质网膜上。GhPK1在种子和纤维中的表达水平较其它组织高。另外,GhPK1的表达量在受到盐胁迫后上升,说明GhPK1可能参与了盐胁迫反应。     

水肥调控对二月兰和后茬花生养分累积及土壤肥力的影响

摘 要：田间试验条件下，研究不同水肥处理对二月兰生长，及其翻压后后茬花生产量和养分累积的变化。结果表明，灌溉和施肥可显著促进二月兰生长。在绿肥季，不论施肥与否，灌溉处理均可显著提高二月兰的生物量和N、P、K养分含量， NPW（绿肥季施氮磷肥和灌溉）和CKW处理（绿肥季不施肥，只进行灌溉处理）的二月兰生物量和N、P、K养分含量分别比相应的未灌溉处理提高了66.47%和63.97%、76.95%和32.36%、88.31%和9.80%、21.71%和15.56%。二月兰翻压的养分还田量为91.04～260.23 kg/hm2，约占花生季化肥总养分的27.59%～78.86%。与冬闲处理（CF）相比，不同施肥和灌溉处理的绿肥翻压均促进了花生产量和养分累积，及土壤养分含量的提高，其中以EN处理的提升效果最明显。周年等养分条件下，花生季35.00%氮和/或42.86%磷肥料前移至绿肥季，可明显促进绿肥养分还田量的增加，后茬花生产量不同程度增加（增幅22.82%～41.18%）。综上，在适量灌溉和施肥条件下，二月兰生物量明显增加，进而促进后茬花生产量增加及养分累积。研究结果可为我国绿肥农田应用及化肥减施提供数据支撑和实践依据。     

4种水生植物深度净化村镇生活污水厂尾水效果研究

通过设置动态模拟试验,持续进水、出水条件下分析比较了漂浮植物凤眼莲和水浮莲、沉水植物轮叶黑藻和挺水植物黄菖蒲对村镇生活污水厂(一级A标准)尾水深度净化效果,筛选出具有去污效果优势的水生植物,为优化水生植物生态修复工程技术在尾水深度净化中的应用提供依据。结果表明:经水生植物深度净化后,尾水水质得到明显改善,漂浮植物凤眼莲和水浮莲对尾水氮、磷的净化效果优于挺水植物黄菖蒲和沉水植物轮叶黑藻。试验周期内,污水厂尾水总氮、总磷和高锰酸盐指数(CODMn)平均浓度为12.22 mg?L-1、0.38mg?L-1和3.88 mg?L-1,凤眼莲、水浮莲、轮叶黑藻、黄菖蒲和对照各系统的总氮平均去除率分别为46.25%、45.74%、43.41%、38.39%和29.22%,总磷去除率分别为36.84%、34.21%、31.58%、28.95%和26.32%,CODMn去除率分别为42.27%、30.93%、32.47%、32.47%和37.89%。凤眼莲、水浮莲、黄菖蒲和轮叶黑藻生物量净增长率分别为550.5%、418.8%、210.6%和80.3%,凤眼莲生物量净增率最大。各处理系统内凤眼莲、水浮莲、黄菖蒲和轮叶黑藻对尾水氮富集量分别为7.36 g、2.33 g、5.12 g和4.46 g,对磷的富集量分别为0.60 g、0.19 g、0.33 g和0.78 g,凤眼莲富集氮能力优于另外3种水生植物,轮叶黑藻磷富集量高于另外3种水生植物。凤眼莲、水浮莲、黄菖蒲和轮叶黑藻植株吸收作用对尾水总氮去除的表观贡献率分别为15.29%、4.90%、11.17%和11.34%,对尾水总磷去除的表观贡献率分别为50.34%、17.17%、35.24%和76.34%。因此,可利用漂浮植物凤眼莲和沉水植物轮叶黑藻立体复合种养的方式深度净化生活污水厂尾水。     

白粉菌(Erysiphe graminis)侵染诱导表达的小麦糖苷水解酶基因TaGlc2的克隆与鉴定

真菌病害是世界小麦生产中的最重要的病害之一。目前,在小麦中已经鉴定出一批小麦病菌侵染诱导基因,包括病程相关基因和抗真菌水解酶基因（葡聚糖酶基因和几丁质酶基因）。最近的研究表明,植物1,3-β-葡聚糖酶参与对真菌侵染的防卫。本研究从小麦cDNA文库中克隆出一个编码小麦1,3-β-葡聚糖酶的新基因,命名为 TaGlc2。该基因编码的氨基酸序列与糖苷水解酶基因家族17高度同源。采用实时定量PCR分析方法对TaGlc2基因在白粉菌侵染(Erysiphe graminis)后小麦叶片中的表达模式进行研究,发现TaGlc2基因在白粉菌侵染6 h后表达明显增强,至24 h达到峰值,说明该基因受白粉菌侵染诱导表达。还获得了TaGlc2基因5'上游调控区序列,发现存在与病菌侵染响应有关的顺式元件。小麦TaGlc2基因在小麦白粉病抗性上可能具有重要作用。     

东方粘虫中肠V-ATP酶A亚基突变体TSCA的原核表达及复性纯化

根据得到的东方粘虫[Mythimna separate(Walker)]中肠V-ATPase A亚基(VHA-A)基因(VATPA),预测其三维结构,构建定点突变并复性。利用Swiss-Model在线预测VHA-A三维结构,确定突变位点,通过重折叠PCR(Overlap-PCR)技术构建突变基因并将其整合到pET-22b(+)载体上,得到重组质粒pET-22b(+)-TSCA,转入大肠杆菌BL21中表达,最后通过亲和层析纯化包涵体蛋白质并进行透析复性。结果表明,成功构建突变基因和融合表达载体,融合蛋白质以包涵体形式表达,并得到大量纯化的可溶性蛋白。