脂肪储存小滴蛋白5 重组腺病毒的制备

目的:利用AdEasy腺病毒表达系统构建含有小鼠脂肪储存小滴蛋白5(LSDP5)基因的重组腺病毒。方法:从小鼠肝脏cDNA克隆出LSDP5基因全长,克隆至pMD18-T载体中,酶切测序。回收酶切产物,连接到腺病毒穿梭载体pShuttle-CMV,构建pShuttle-CMV-LSDP5重组质粒,经PmeI酶切线性化后转化至含有腺病毒骨架质粒pAdEasy-1的BJ5183中。筛选阳性克隆,提取重组质粒,PacI酶切线性化并转染AD293细胞进行包装,提取病毒DNA,鉴定重组病毒并检测病毒滴度。结果:LSDP5基因克隆经测序证实与Genebank公布一致,双酶切重组pMD18-T载体得到1400 bp左右的片段。重组穿梭载体经Kpn I和Sal I双酶切后得到预期片段。PacI酶切得到30 Kb大片段和4.5 Kb小片段。转染AD293细胞后收集病毒,经PCR鉴定,获得理想的目的片段。取病毒上清反复感染AD293细胞以扩增病毒,最后所得病毒滴度为2.5×109pfu/ml。结论:成功构建了携带脂肪储存小滴蛋白5基因的重组腺病毒载体,为进一步研究LSDP5基因功能奠定基础。     

80%门静脉结扎后肝脏组织中MMP-9 的表达与肝再生作用 的相关性研究

目的:探讨基质金属蛋白酶-9(MMP-9)在大鼠80%门静脉分支结扎模型中大鼠增生肝脏组织中的表达及其与肝再生作用的关系。方法:健康SD雄性大鼠48只,随机平均分成假手术对照组(Sham)和门静脉结扎实验组(PVL)。观察术后1、3、7和14d保留侧肝叶重量/体重比值;下腔静脉采血后检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)值的变化;光镜下观察保留侧肝脏组织的病理形态变化;用免疫组化法检测增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)的表达,用免疫印记法检测MMP-9的表达,并进行统计分析。结果:80%门静脉分支结扎后,结扎侧肝叶呈进行性萎缩,保留侧肝叶重量/体重比值逐渐增加,7d达"平台期";与对照组明显不同,PVL组的ALT、AST的值在1h达到高峰,7d后回到正常水平;保留侧肝脏组织中PCNA阳性细胞计数与对照组比较,3d开始表达增强(P<0.05),7d以后逐渐恢复至正常水平(P>0.05);保留侧肝叶MMP-9蛋白的表达在术后3d开始增加,术后7d达到高峰。结论:MMP-9蛋白的表达在80%门静脉结扎后大鼠肝再生过程中发挥重要作用。     

The structural features of Trask that mediate its anti-adhesive functions

Trask/CDCP1 is a transmembrane protein with a large extracellular and small intracellular domains. The intracellular domain (ICD) undergoes tyrosine phosphorylation by Src kinases during anchorage loss and, when phosphorylated, Trask functions to inhibit cell adhesion. The extracellular domain (ECD) undergoes proteolytic cleavage by serine proteases, although the functional significance of this remains unknown. There is conflicting evidence regarding whether it functions to signal the phosphorylation of the ICD. To better define the structural determinants that mediate the anti-adhesive functions of Trask, we generated a series of deletion mutants of Trask and expressed them in tet-inducible cell models to define the structural elements involved in cell adhesion signaling. We find that the ECD is dispensable for the phosphorylation of the ICD or for the inhibition of cell adhesion. The anti-adhesive functions of Trask are entirely embodied within its ICD and are specifically due to tyrosine phosphorylation of the ICD as this function is completely lost in a phosphorylation-defective tyrosine-phenylalanine mutant. Both full length and cleaved ECDs are fully capable of phosphorylation and undergo phosphorylation during anchorage loss and cleavage is not an upstream signal for ICD phosphorylation. These data establish that the anti-adhesive functions of Trask are mediated entirely through its tyrosine phosphorylation. It remains to be defined what role, if any, the Trask ECD plays in its adhesion functions.     

滇龙胆不同部位裂环烯醚萜类物质含量比较分析

采用高效液相色谱法,对云南临沧云县滇龙胆不同部位龙胆苦苷、獐牙菜苦苷、当药苷含量进行分析.结果显示:(1)龙胆苦苷、獐牙菜苦苷、当药苷在滇龙胆根、茎、叶、花中均有分布;龙胆苦苷、獐牙菜苦苷在根部含量较高,分别为(35.88±5.89)mg/g和(1.35±0.33)mg/g;当药苷在花部含量最高,为(0.97±0.24)mg/g.(2)不同部位有效成分间具有显著(P<0.05)或极显著(P<0.01)的相关性.(3)相同海拔条件下,不同栽培方式下滇龙胆根部龙胆苦苷含量差异不显著(P>0.05),但茎部、叶部獐牙菜苦苷含量差异显著(P<0.05).     

游离植皮联合负压封闭引流治疗骨科创面的临床研究

目的:观察游离植皮联合负压封闭引流(vacuumsealingdrainage,后文简称VSD)对骨科创面的疗效,并与植皮后传统加压包扎相比较,为临床实践提供更好的治疗方法。方法:对广州中医药大学第一附属医院创伤骨科2008年3月至2010年2月收治的65例感染创面病例采取手术清创后予VSD引流,合理应用抗生素,创面感染得到控制后,创面干净,肉芽生成良好,外露的肌腱、骨膜表面有新鲜的肉芽组织覆盖,达到植皮的要求后,随机分成两组,其中30例(实验组)采用游离植皮联合VSD法闭合创面,35例(对照组)采用游离植皮加压包扎植皮区,对两组术后的平均换药次数、创面平均愈合时间、植皮成活率情况、平均住院时间(植皮后)、平均抗生素应用次数(植皮后)进行统计学分析,采用t检验和卡方检验,对此两种方法进行评价。结果:植皮联合VSD组与植皮加压包扎组,在平均换药次数、创面平均愈合时间、植皮成活率、平均住院时间(植皮后)、平均抗生素应用次数(植皮后)的对比,有显著性差异(P<0.05)。结论:创面达到游离植皮条件后,游离植皮联合VSD负压引流可以促使皮片黏附,保持创面洁净,避免皮下渗液积聚,有利于皮片的存活,与植皮加压包扎组相比,减少了平均换药次数,缩短创面平均愈合时间及平均住院时间(植皮后),减少抗生素平均应用次数,提高了植皮成活率,说明游离植皮联合VSD组优于游离植皮加压包扎组,游离植皮联合VSD法治疗骨科创面有显著疗效。该手术方法操作简单,术后护理方便,是一种较理想的植皮后的固定方法,有利于创面的愈合,值得临床推广应用。     

流行性出血热病毒R22株cDNA克隆及其特异性鉴定

用家鼠型流行性出血热病毒R22株RNA,经polyA接尾,以Oligo-dT做引物,合成cDNA。用pUC18为载体转染E.coli Mc1061,建立cDNA克隆。再经菌落杂交,选择病毒特异性的5个阳性克隆制成缺口翻译探针,与病毒RNA3个片段进行反杂交,确定RNA片段的特异性。结果表明,3个克隆为中(M)片段的cDNA,另两个分别为大(L)和小(S)片段cDNA。核苷酸序列分析证明,克隆的DNA中含病毒特异的核苷酸序列。     

中国东北香蒲属植物花粉的营养价值及开发潜力分析

香蒲属（Typha）系香蒲科（Typhaceae）一单属,水生或沼生。全国共11种［‘］,东北地区约7种。香蒲属植物是重要的经济植物和珍贵的药材,香蒲花粉用于止血,效果很好;叶片可用于纺织、造纸等;幼叶基部和根状茎先端可作蔬菜食用;雌花序成熟后可作枕芯和坐垫的填充物．从营养学角度来看,目前对香蒲花粉还缺乏较系统的研究和开发利用．作者着重从香蒲花粉的营养价值的角度来展示其具有巨大开发潜力的野生资源．l材料与方法选取三种香蒲,即宽叶香蒲TyphalatdoliaL．（No．l）、水烛TangustifoliaL．（No．2）和无苞香蒲Tla。anniiLe…     

Climatic Factors Drive Population Divergence and Demography: Insights Based on the Phylogeography of a Riparian Plant Species Endemic to the Hengduan Mountains and Adjacent Regions

Quaternary climatic factors have played a significant role in population divergence and demography. Here we investigated the phylogeography of Osteomeles schwerinae, a dominant riparian plant species of the hot/warm-dry river valleys of the Hengduan Mountains (HDM), Qinling Mountains (QLM) and Yunnan-Guizhou Plateau (YGP). Three chloroplast DNA (cpDNA) regions (trnD-trnT, psbD-trnT, petL-psbE), one single copy nuclear gene (glyceraldehyde 3-phosphate dehydrogenase; G3pdh), and climatic data during the Last Interglacial (LIG; c. 120–140 ka), Last Glacial Maximum (LGM; c. 21 ka), and Current (c. 1950–2000) periods were used in this study. Six cpDNA haplotypes and 15 nuclear DNA (nDNA) haplotypes were identified in the 40 populations of O. schwerinae. Spatial Analysis of Molecular Variance, median-joining networks, and Bayesian phylogenetic trees based on the cpDNA and nDNA datasets, all suggested population divergence between the QLM and HDM-YGP regions. Our climatic analysis identified significant heterogeneity of the climatic factors in the QLM and HDM-YGP regions during the aforementioned three periods. The divergence times based on cpDNA and nDNA haplotypes were estimated to be 466.4–159.4 ka and 315.8–160.3 ka, respectively, which coincide with the time of the weakening of the Asian monsoons in these regions. In addition, unimodal pairwise mismatch distribution curves, expansion times, and Ecological Niche Modeling suggested a history of population expansion (rather than contraction) during the last glaciation. Interestingly, the expansion times were found being well consistent with the intensification of the Asian monsoons during this period. We inferred that the divergence between the two main lineages is probably caused by disruption of more continuous distribution because of weakening of monsoons/less precipitation, whilst subsequent intensification of the Asian monsoons during the last glaciation facilitated the expansion of O. schwerinae populations.     

消化道氮能神经的研究进展

人体消化道的神经构成复杂、功能多样,向来是学术界的研究热点之一。与其相关的新观点、新概念、新物质的报道层出不穷。20世纪末期,基于对一氧化氮(NO)生物学作用广泛而深入地研究,人们发现并明确了NO是一种新型神经递质。由于此类以NO为递质的神经具有重要的生物学效应,学术界提出了氮能神经的概念从而为我们深入了解非肾上腺素能非胆碱能神经开辟了全新的视角,同时也彰显出氮能神经在生物体中潜在的重大作用。本文将针氮能神经在胃、肠道、食管等主要消化道器官的分布情况、对消化道正常生理功能调控、以及与相关消化道疾病病理联系等方面展开综述,并设想了氮能神经可能基于一氧化氮合酶的各种异构体的相关性而在消化道正常、疾病两态之间的平衡机制中存在某种作用和联系。     

Involvement of a Magnaporthe grisea serine/threonine kinase gene, MgATG1, in appressorium turgor and pathogenesis

We isolated an MgATG1 gene encoding a serine/threonine protein kinase from the rice blast fungus Magnaporthe grisea. In the DeltaMgatg1 mutant, in which the MgATG1 gene had been deleted, autophagy was blocked; the mutant also showed fewer lipid droplets in its conidia, lower turgor pressure of the appressorium, and such defects in morphogenesis as delayed initiation and slower germination of conidia. As a result of lower turgor pressure of the appressorium, the DeltaMgatg1 mutant lost its ability to penetrate and infect the two host plants, namely, rice and barley. However, normal values of the parameters and infective abilities were restored on reintroducing an intact copy of the MgATG1 gene into the mutant. Autophagy is thus necessary for turnover of organic matter during the formation of conidia and appressoria and for normal development and pathogenicity in M. grisea.