cAMP 参与烟草悬浮细胞的ABA信号转导

ABA诱导型启动子(rd29A)重组到报告基因(GUS)的上游构建表达载体.通过农杆菌介导转化烟草,获得转基因植株.将转基因植株诱导愈伤组织,建立稳定、均一的转rd29A-GUS融合基因的悬浮培养细胞系.用ABA处理悬浮细胞24 h后GUS活性显著升高,说明外源ABA能够诱导rd29A启动子的表达,获得了用于ABA信号转导研究的实用细胞系.在ABA激活表达的细胞介质中加入尼克酰胺(cADPR合成酶的抑制剂)或U73122(PLC抑制剂)只能部分抑制ABA的效应,但如果加入蛋白激酶抑制剂K252a,抑制效果达95%以上.用可跨膜的cAMP的类似物8-Br-cAMP处理细胞发现,它能代替ABA的作用;当介质中加入1 mmol/L IBMX(磷酸二酯酶的抑制剂)增加cAMP的稳定性,发现低浓度的8-Br-cAMP与ABA相同的效应.以上结果表明cAMP参与了烟草悬浮细胞中ABA信号的传递.     

Palmin诱导烟草氧化猝发的机制

为探讨植物对病原微生物的防御机制和激发子启动植物体内的信号转导应答过程 ,本文研究了Phytoph thorapalmi激发子palmin诱导其非寄主亲和性烟草的叶片和悬浮细胞系产生氧化猝发的分子机理。利用生化分析和激光共聚焦显微扫描技术动态观察palmin诱导烟草过敏反应中O·- 2 和H2 O2 的形成、胞间转移及引起细胞死亡的特性。结果表明 :palmin诱导激活了烟草细胞内NADPH氧化酶 ,产生大量的O·- 2 ;O·- 2 在SOD催化下迅速转变成H2 O2 ,并且H2 O2 在一定范围的细胞间转移和积累 ,最后诱发烟草细胞的过敏性坏死反应。palmin诱导氧化猝发过程还有Ca2 和蛋白激酶的参与。     

植物逆境胁迫耐受性功能基因组研究进展

为了更加高效地利用基因工程技术提高植物对逆境胁迫的耐受性,需要在全基因组水平上对植物逆境胁迫耐受性的复杂机制进行整合性研究.植物逆境胁迫耐受性功能基因组的研究可概括为：利用胁迫特异性的表达序列标签(EST)及cDNA微阵列（或基因芯片）技术筛选与胁迫相关的候选基因,然后利用反向遗传学等技术对候选基因的功能进行研究,利用酵母双杂交、正向遗传学等技术对基因及基因产物间的相互关系进行研究.通过这些研究可以全面地了解植物对胁迫（渗透、干旱、极端温度）响应的复杂机制和相互作用以及相应的信号转导途径,从而为更加高效地利用基因工程技术提高植物对逆境胁迫的耐受性奠定基础.     

The Arabidopsis inositol 1,3,4-trisphosphate 5/6 kinase, AtItpk-1, is involved in plant photomorphogenesis under red light conditions, possibly via interaction with COP9 signalosome.

The COP9 signalosome (CSN) plays important roles in multifaceted cellular processes. Study has shown that inositol 1,3,4-trisphosphate 5/6 kinase (5/6 kinase) interacts with CSN in mammalian cells. However, the biological function of the interaction still remains unknown. Here, we report that the Arabidopsis inositol 1,3,4-trisphosphate 5/6 kinase (AtItpk-1) is also associated with CSN and involved in photomorphogenesis under red light (RL) conditions, as demonstrated by co-immunoprecipitation of AtItpk-1 with CSN and characterization of the atitpk-1 mutants. Expression analysis showed that AtItpk-1 had the same sub-cellular localization and organ expression pattern as CSN. Furthermore, autophosphorylation analysis showed that AtItpk-1 has protein kinase activity. Under RL, the atitpk-1 mutants exhibited phenotype slightly similar with that of the csn mutants, indicating that 5/6 kinase might be involved in the same developmental pathway as CSN. This study suggests that AtItpk-1 may function as a protein kinase that is involved in photomorphogenesis possibly via interaction with COP9 signalosome under red light.     

豌豆根胞质V1-ATPase的鉴定

利用免疫印迹、免疫电镜和ATP水解活性的测定对豌豆(Pisum sativum L.)根细胞胞质中V1-ATPase复合物的存在进行鉴定.用兔抗绿豆V-type H+-ATPase 的A、B亚基的抗体进行的immuno-blotting和胶体金电镜结果都表明,胞质中存在有A、B亚基.活性测定结果进一步表明胞质具有ATP水解活性.这些结果说明豌豆根胞质具有有活性的V1-ATPase复合物.这是首次直接证明植物中有胞质V1-ATPase的存在.     

Stochastic dynamics of actin filaments in guard cells regulating chloroplast localization during stomatal movement

Actin filaments and chloroplasts in guard cells play roles in stomatal function. However, detailed actin dynamics vary, and the roles that they play in chloroplast localization during stomatal movement remain to be determined. We examined the dynamics of actin filaments and chloroplast localization in transgenic tobacco expressing green fluorescent protein (GFP)-mouse talin in guard cells by time-lapse imaging. Actin filaments showed sliding, bundling and branching dynamics in moving guard cells. During stomatal movement, long filaments can be severed into small fragments, which can form longer filaments by end-joining activities. With chloroplast movement, actin filaments near chloroplasts showed severing and elongation activity in guard cells during stomatal movement. Cytochalasin B treatment abolished elongation, bundling and branching activities of actin filaments in guard cells, and these changes of actin filaments, and as a result, more chloroplasts were localized at the centre of guard cells. However, chloroplast turning to avoid high light, and sliding of actin fragments near the chloroplast, was unaffected following cytochalasin B treatment in guard cells. We suggest that the sliding dynamics of actin may play roles in chloroplast turning in guard cells. Our results indicate that the stochastic dynamics of actin filaments in guard cells regulate chloroplast localization during stomatal movement.     

Overexpression of the Arabidopsis ��-expansin gene AtEXPA1 accelerates stomatal opening by decreasing the volumetric elastic modulus

Guard cell walls of stomata are highly specialized in plants. Previous research focused on the structure and anatomy of guard cell walls, but little is known about guard cell regulation during stomata movement. In this work, we investigate the possible biological role of the Arabidopsis expansin gene AtEXPA1 in stomatal opening. The AtEXPA1 promoter drove the expression of the GUS reporter gene specifically in guard cells. Light-induced stomatal opening was accelerated in 35S::AtEXPA1 lines, whereas the anti-AtEXPA1 antibody decelerated light-induced stomatal opening. The inhibition of the anti-AtEXPA1 antibody on stomatal opening was largely dependent on the environmental pH. The volumetric elastic modulus (ε) was measured as an indicator of changes in the cell wall. The ε value of guard cells in 35S::AtEXPA1 lines was smaller than in the wild types. The putative role of AtEXPA1 as controller of stomatal opening rate and its regulation are discussed.     

解剖型锁定钢板治疗Pilon骨折的临床疗效分析

目的:探讨解剖型锁定钢板治疗Pilon骨折的临床疗效和安全性。方法:选择2009年1月~2012年12月采用解剖型锁定钢板内固定治疗30例Pilon骨折患者为研究对象,所有患者在无明显手术禁忌情况下,均行开放复位钢板内固定术。术后采用Mazur评分系统评价手术效果。结果:本组全部患者均获得随访,骨折平均愈合时间为4.1个月;参照Mazur踝关节功能评分系统:优14例,良13例,可2例,差1例,优良率90%。仅1例开放性Pilon骨折患者发生皮肤感染坏死,局部缺损,经清创、VSD覆盖负压吸引,待新鲜肉芽组织形成后,行带蒂皮瓣移植术,愈合良好。结论:解剖型锁定钢板的解剖形态好,成角稳定,用于治疗Pilon骨折能够恢复关节面的完整性、稳定性,减少并发症的发生,临床效果好,安全性高。     

Influence of Low Temperature on Spatial-temporal Changes of Ca2+ in Winter or Spring Wheat

Stained with Fluo-3/AM and investigated under the laser scanning confocal microscope (LSCM), the spatial-temporal changes of ［Ca2+］cyt(the free Ca2+ concentration in the cytoplasm) in the protoplasts of mesophyll cells of wheat (Triticum aestivum L.) cultivars with different cold-hardiness under resting and temperature decreasing conditions were compared. The results showed that under the resting condition, the fluorescence intensities in the protoplasts of both cold-sensitive and cold-resistant wheat manifested no significant changes, implying that ［Ca2+］cyt could keep at a stable level under the resting condition although different wheat cultivars were different. However, different dynamics appeared with temperature decreasing from 15℃ to 2℃. In particular, for the cold-resistant wheat, ［Ca2+］cyt level at first increased, followed by drop to the resting level at 2℃, and then increased further when the temperature was below 2℃. On the contrary, the ［Ca2+］cyt level of the cold-sensitive spring wheat increased continuously to the maximal level within the whole range of temperature changes. Based on the above observations, it could be reasonably inferred that different dynamics of ［Ca2+］cyt determines decisively different cold acclimation abilities of plants. Furthermore, the results provide a new supporting evidence for the hypothesis that Ca2+ plays a key role as a primary physiological transducer upon chilling”.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细