云南高黎贡山和怒山外担菌属和隔担菌属真菌

2005至2011年,在云南高黎贡山和怒山进行了野外科学考察,采集了大量真菌标本,经过研究,发现外担菌属18种,隔担菌属11种真菌,研究的标本保藏在中国科学院微生物研究所菌物标本馆.     

Rapid apoplastic pH measurement in Arabidopsis leaves using a fluorescent dye

In plants, the extracellular space (apoplast) is one of the main places where exchange of molecules occurs between cells. Not only is this compartment involved in the storage of multiple metabolites and ions, including calcium and protons, but it also plays a role in the transmission of signaling molecules for cell-to-cell communication. It has recently been shown multiple times that these two aspects are linked and can influence each other. In particular, apoplast pH was shown as a primary regulator of auxin (IAA) transport in Arabidopsis thaliana. To prove the role of apoplastic pH, we have developed a protocol for apoplastic fluid extraction from Arabidopsis leaves, followed by pH determination using the 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) fluorescent dye. This technique successfully allows one to monitor apoplastic pH variations among different plant lines and to link changes in apoplastic pH to cellular responses in the plant.     

RNA干扰茄病镰刀菌ALP基因及其基因沉默菌株的制备

目的构建RNA干扰质粒载体抑制茄病镰刀菌碱性丝氨酸蛋白酶(ALP)基因,通过检测ALP的表达及酶活性变化筛选出基因沉默菌株。方法构建2个ALP的干扰载体,转化茄病镰刀菌孢子,获得ALP基因沉默菌株△ALP1、△ALP2,通过RT-PCR检测△ALP1、△ALP2的ALP基因mRNA变化,并通过蛋白琼脂廓清率培养基检测ALP酶活性变化,对所得菌株的毒力进行判断。结果 RNA干扰后所获基因沉默菌株中,ALP的mRNA表达量显著低于标准茄病镰刀菌株(F=184.67,P<0.01),其中ΔALP2抑制效果较好、酶活性显著低于标准菌株及△ALP1(q=5.276、5.463,P<0.01)。结论通过LiAc转化法对茄病镰刀菌ALP进行RNA干扰,可有效抑制茄病镰刀菌ALP的表达;ALP基因沉默茄病镰刀菌株的获取,为进行动物体内实验、深入研究ALP在茄病镰刀菌性角膜炎发病机制中的作用、探索新型治疗方法提供思路。     

藜磷酸肌醇5-磷酸酶基因(CaP5P)的表达分析和胁迫响应检测

植物通过多种信号途径感知外界环境变化,从而引发一系列的信号级联反应,其中磷脂信号途径起着重要的作用。本研究从藜盐胁迫差减文库中获得了一个EST05序列,利用RT-PCR检测了其在非生物胁迫下表达趋势,显示EST05序列在非生物胁迫下的表达有升高。随后利用RACE技术获得其全长编码序列,测序结果显示其与蓖麻(Ricinus communis) 的磷酸肌醇5-磷酸酶基因(P5P)的同源性达56%,遂将其命名为CaP5P。最后过量表达CaP5P在拟南芥中初步验证了功能,转基因植株表现对盐胁迫耐受性降低的趋势,呈现负调控的特征。本研究对磷酸肌醇5-磷酸酶基因功能的初步研究为磷脂信号途径中负调控组分的分析提供了参考依据。     

额底纵裂入路手术对鞍上第三脑室底垂体瘤的疗效分析

目的：分析经额底纵裂入路治疗鞍上第三脑室底垂体瘤的疗效,探讨其临床适用性。方法：选择从2011年1月~2013年1月与我院行额底纵裂入路手术治疗的30例鞍上第三脑室底垂体瘤的患者,术中行单侧额或双侧额弧形切口,根据术中所见肿瘤位置,由终板、视神经一颈内动脉等存在的生理间隙处切除肿瘤,观察所有患者的手术疗效。结果：所有患者术中可见肿瘤位于鞍内鞍上,部分或全部突入第三脑室底,其中有6例患者伴有脑积水。术中肿瘤全切23例,次全切5例,大部切除2例,无手术死亡病例。术前25例患者视力减退,术后23例患者视力均获得不同程度改善,仅1例暂无明显变化。术后19例出现电解质紊乱,患者经治疗后均已纠正;12例出现不同程度的尿崩症,给予患者药物治疗后,病情得到缓解。术后随访6个月,23例肿瘤全切患者病灶无复发,另外4例次全切者病灶也无明显变化,仅1例次全切和2例大部切除患者于术后行伽马刀再次治疗。结论：经额底纵裂入路治疗鞍上第三脑室底垂体瘤可以达到视野清晰,直观下进行肿瘤切除,手术效果好,并发症较少,适合临床长期推广应用。     

地佐辛联合丙泊酚用于无痛肠镜诊疗的临床观察

目的：观察地佐辛联合丙泊酚用于无痛肠镜诊疗的有效性和安全性。方法：90例患者随机分为三组：地佐辛组（D组）、芬太尼组（F组）和丙泊酚组（P组）,观察患者给药前（T0）、插镜时（T1）、停药时（T2）和睁眼时（T3）的收缩压（SBP）、舒张压（DBP）、心率（HR）和血氧饱和度（SpO2）;手术时间及术中丙泊酚的用药总量、苏醒时间和离院时间;停药后20 min内Ramsay评分;术中和术后的不良反应,术后患者和术者的满意度。结果：D组的血流动力学指标比F组和P组稳定;P组的丙泊酚用量明显大于F组和D组（P〈0.01）;P组的苏醒时间长于F组（p〈0.05）和D组（P〈0.01）,且离院时间也明显长于F组和D组;停药10 min时,D组的Ramsay评分小于P组（P〈0.05）,停药15 min时,D组和F组的Ramsay评分均小于P组（P〈0.05）。结论：地佐辛联合丙泊酚用于无痛胃肠镜诊疗更安全有效,值得临床推广。     

A Novel Balanced-Lethal Host-Vector System Based on glmS

During the last decade, an increasing number of papers have described the use of various genera of bacteria, including E. coli and S. typhimurium, in the treatment of cancer. This is primarily due to the facts that not only are these bacteria capable of accumulating in the tumor mass, but they can also be engineered to deliver specific therapeutic proteins directly to the tumor site. However, a major obstacle exists in that bacteria because the plasmid carrying the therapeutic gene is not needed for bacterial survival, these plasmids are often lost from the bacteria. Here, we report the development of a balanced-lethal host-vector system based on deletion of the glmS gene in E. coli and S. typhimurium. This system takes advantage of the phenotype of the GlmS⁻ mutant, which undergoes lysis in animal systems that lack the nutrients required for proliferation of the mutant bacteria, D-glucosamine (GlcN) or N-acetyl-D-glucosamine (GlcNAc), components necessary for peptidoglycan synthesis. We demonstrate that plasmids carrying a glmS gene (GlmS⁺p) complemented the phenotype of the GlmS⁻ mutant, and that GlmS⁺p was maintained faithfully both in vitro and in an animal system in the absence of selection pressure. This was further verified by bioluminescent signals from GlmS ⁺pLux carried in bacteria that accumulated in grafted tumor tissue in a mouse model. The signal was up to several hundred-fold stronger than that from the control plasmid, pLux, due to faithful maintenance of the plasmid. We believe this system will allow to package a therapeutic gene onto an expression plasmid for bacterial delivery to the tumor site without subsequent loss of plasmid expression as well as to quantify bioluminescent bacteria using in vivo imaging by providing a direct correlation between photon flux and bacterial number.     

The Australian giant fleas Macropsylla Rothschild, 1905 (Siphonaptera: Macropsyllidae: Macropsyllinae), their identification,evolution, ecology,and conservation biology

Systematic Parasitology - Macropsylla Rothschild, 1905 is an endemic Australian flea genus represented by two species: M. hercules Rothschild, 1905 and M. novaehollandiae Hastriter, 2002. However,...     

碳源影响水稻根际假单胞菌PA1201藤黄绿菌素的生物合成

【背景】假单胞菌PA1201是一株水稻根际促生菌,其产生的次生代谢物藤黄绿菌素(pyoluteorin,Plt)能够有效抑制多种植物病原真菌和细菌的生长,但在常规培养条件下Plt产量极低。【目的】研究碳源对Plt生物合成的影响,为提高Plt的产量以及应用提供理论基础。【方法】将基本培养基(minimal medium,MM)中甘露醇替换为不同的碳源及碳源组合作为PA1201的培养基,生长过程中不同时间点取样提取Plt,利用高效液相色谱(HPLC)法分析Plt的产量变化。【结果】建立了基于HPLC定性和定量检测Plt的方法;比较了PA1201菌株在不同培养基中菌株生长和Plt的产量,发现果糖和甘露醇促进Plt生物合成;果糖和甘露醇对Plt生物合成没有增效作用;在含有甘露醇或果糖作为唯一碳源的培养基中,添加葡萄糖或琥珀酸抑制Plt生物合成。【结论】果糖和甘露醇促进水稻根际假单胞菌PA1201合成藤黄绿菌素,这为提高藤黄绿菌素的生物合成效率和促进藤黄绿菌素的应用奠定了基础。