He-Ne激光对小麦ROP GTPase经增强UV-B辐射造成的损伤的修复研究

采用He-Ne激光辐照对增强UV-B辐射后小麦幼苗ROP GTPase损伤修复作用进行了研究。采用了SDS-PAGE电泳法检测各组ROP GTPase的含量和激光共聚焦显微镜对小麦微丝进行FITC荧光强度标记的测定。研究结果表明：经增强UV-B辐射后,小麦幼苗的Rop GTPase含量降低,LCSM扫描细胞原生质体形状发生改变,细胞骨架受到破坏,其被标记的荧光变暗,强度减弱,在整个UV-B处理期间均低于对照组（CK）,再以He-Ne激光处理后,其含量,形状和强度均有所提高,但仍低于对照组。由此说明,UV-B辐射能使小麦幼苗的Rop GTPase含量下降,微丝骨架受到破坏,一定剂量的激光对UV-B辐射后小麦的蛋白含量和细胞骨架有一定修复作用。Rop GTPase参与了微丝骨架重组的过程。     

菰黑粉菌的分离鉴定及其发酵液中植物激素的检测

在显微镜下观察孢子形态,并观察单菌落的形态和菌株的微观形态,PCR扩增其ITS-5.8S rDNA序列,测序并在NCBI中进行同源比较,确定其种属。液体培养该菌株,通过高效液相色谱法检测发酵液中植物激素。结果表明：用菌落形态与孢子形态鉴定和分子生物学鉴定的方法,对茭白中分离的一个菌株鉴定为菰黑粉菌,且在其发酵液中检测到植物激素IAA、ABA和GA3,其中IAA含量为0.1306mg/L,ABA含量为0.01367mg/L。     

CSPGs在大鼠脊髓损伤后的表达及其与GFAP的关系

目的:观察脊髓损伤后CSPGs的表达及其与GFAP的关系。方法:成年雄性SD大鼠25只,随机分为对照组和损伤组,损伤组分脊髓挤压损伤后0h、72h、1w、4w组,运用免疫荧光双重染色方法观察CSPGs与GFAP的表达。结果:挤压伤后损伤部位的CSPGs和GFAP的表达均增高,但二者的变化趋势并不一样。其中CSPGs从损伤后表达开始增高,此后一直增加,并在1w至4w时逐渐稳定,主要分布逐渐集中于损伤部位;星形胶质细胞的免疫反应也逐渐增加,其分布逐渐集中于损伤区域的边缘,逐渐形成胶质瘢痕界膜。损伤1w至4w,损伤区域内几乎没有了星形胶质细胞表达,但仍留有大量的CSPGs。结论:早期抑制星形胶质细胞分泌CSPGs,可以防止在损伤部位沉积大量的CSPGs,从而减小其对再生纤维的抑制作用。     

基质及水位对入侵种互花米草生长的影响

为了揭示基质和水位对互花米草种群扩张过程的影响,通过室内模拟试验分析了不同基质和地下水位对互花米草幼苗存活率、高生长和生物量及分配等表型参数的影响。结果表明:基质、水位及其交互作用对互花米草幼苗生长影响显著;基质类型和地下水位的交互作用显著影响幼苗的存活率;同种基质中地下水位的降低显著促进了幼苗根长的增加;由于不同基质持水力和营养的差异,沙中生长的幼苗随地下水位的降低地上部分表现出显著的生长抑制,而沙土和壤土中的幼苗在中等水位生长最好;沙中幼苗根状茎的长度显著低于沙土和壤土,而随水位的降低,沙土和壤土根茎长度显著下降。由此可以推论,互花米草种群扩张强度因生境而异,在沙土和壤土基质中互花米草可能表现出更强的入侵性和扩散能力。     

潜流型菖蒲人工湿地不同C/N对污染物的去除效率

选取炉渣和砾石为基质,以无植被为对照,分别设置低、中、高浓度的3个碳水平(C1、C2、C3)和3个氮水平(N1、N2、N3)处理,研究潜流型菖蒲人工湿地在不同C/N下净化生活污水中COD、总氮(TN)、总磷(TP)的效果。结果表明,在不同C/N下,菖蒲人工湿地对污水中COD、TN的去除效果显著高于无植被的人工湿地,菖蒲植被能增加人工湿地COD去除率10.53%,增加TN去除率6.73%;而对于TP的去除,有无植被无显著差异。随着进水N、P浓度及C/N的变化,菖蒲湿地对COD、TN和TP的去除率分别为67.57%～75.85%、20.91%～56.82%和7.15%～17.78%;同时,菖蒲植株对N、P的积累量也相应的变化,其地上部的N、P积累量为4.44～14.79和1.11～3.37g.m-2,平均占湿地N、P去除率的6.91%和10.67%;地下部的N、P积累量分别为2.35～5.20和0.74～1.41g.m-2,平均占湿地N、P去除率的2.69%和6.02%。植物地上器官对湿地N、P的积累量大于地下部,有利于通过收割作用去除湿地系统中的N、P。     

Probabilistic Daily ILI Syndromic Surveillance with a Spatio-Temporal Bayesian Hierarchical Model

Background

For daily syndromic surveillance to be effective, an efficient and sensible algorithm would be expected to detect aberrations in influenza illness, and alert public health workers prior to any impending epidemic. This detection or alert surely contains uncertainty, and thus should be evaluated with a proper probabilistic measure. However, traditional monitoring mechanisms simply provide a binary alert, failing to adequately address this uncertainty.

Methods and Findings

Based on the Bayesian posterior probability of influenza-like illness (ILI) visits, the intensity of outbreak can be directly assessed. The numbers of daily emergency room ILI visits at five community hospitals in Taipei City during 2006–2007 were collected and fitted with a Bayesian hierarchical model containing meteorological factors such as temperature and vapor pressure, spatial interaction with conditional autoregressive structure, weekend and holiday effects, seasonality factors, and previous ILI visits. The proposed algorithm recommends an alert for action if the posterior probability is larger than 70%. External data from January to February of 2008 were retained for validation. The decision rule detects successfully the peak in the validation period. When comparing the posterior probability evaluation with the modified Cusum method, results show that the proposed method is able to detect the signals 1–2 days prior to the rise of ILI visits.

Conclusions

This Bayesian hierarchical model not only constitutes a dynamic surveillance system but also constructs a stochastic evaluation of the need to call for alert. The monitoring mechanism provides earlier detection as well as a complementary tool for current surveillance programs.     

Intracranial Orthotopic Allografting of Medulloblastoma Cells in Immunocompromised Mice

Medulloblastoma is the most common pediatric tumor of the nervous system. A large body of animal studies has focused on cerebellar granule neuron precursors (CGNPs) as the cell-of-origin for medulloblastoma^1-4. However, the diverse clinical presentations of medulloblastoma subtypes in human patients (nodular, desmoplastic, classical and large cell/anaplastic), and the fact that medulloblastoma is found in a subset of human patients with no ectopic expression of CGNP marker⁵, suggest that the cellular and molecular origins of medulloblastoma are more complex and far from being completely deciphered. Therefore, it is essential to determine whether there is an alternative medulloblastoma tumor cell-of-origin based on which cell-type specific therapeutic modality can be developed. To this end, intracranial orthotopic allografting of genetically marked tumor cell types followed by subsequent analyses of secondary tumor development in recipients will allow determination of the cellular origin of tumor-initiating cells. Here we describe the experimental protocol for intracranial orthotopic allografting of medulloblastoma cells derived from primary tumor tissue, and this procedure can also be used for transplanting cells from established cell lines.Download video file.^{(31M, mov)}     

Isolation,Enrichment, and Maintenance of Medulloblastoma Stem Cells

Brain tumors have been suggested to possess a small population of stem cells that are the root cause of tumorigenesis. Neurosphere assays have been generally adopted to study the nature of neural stem cells, including those derived from normal and tumorous tissues. However, appreciable amounts of differentiation and cell death are common in cultured neurospheres likely due to sub-optimal condition such as accessibility of all cells within sphere aggregates to culture medium.Medulloblastoma, the most common pediatric CNS tumor, is characterized by its rapid progression and tendency to spread along the entire brain-spinal axis with dismal clinical outcome. Medulloblastoma is a neuroepithelial tumor of the cerebellum, accounting for 20% and 40% of intracranial and posterior fossa tumor in childhood, respectively¹. It is now well established that Shh signaling stimulates proliferation of cerebellar granule neuron precursors (CGNPs) during cerebellar development ^2-4. Numerous studies using mouse models, in which the Shh pathway is constitutively activated, have linked Shh signaling with medulloblastoma ^5-9.A recent report has shown that a subset of medulloblastoma cells derived from Patched1^LacZ/+ mice are cancer stem cells, which are capable of initiating and propogating tumors ¹⁰. Here we describe an efficient method to isolate, enrich and maintain tumor stem cells derived from several mouse models of medulloblastoma, with constitutively activated Shh pathway due to a mutation in Smoothened (¹¹, hereon referred as SmoM2), a GPCR that is critical for Shh pathway activation. In every isolated medulloblastoma tissue, we were able to establish numerous highly proliferative colonies. These cells robustly expressed several neural stem cell markers such as Nestin and Sox2, can undergo serial passages (greater than 20) and were clonogenic. While these cultured tumor stem cells were relatively small, often bipoar with high nuclear to cytoplasmic ratio when cultured under conditions favoring stem cell growth, they dramatically altered their morphology, extended multiple cellular processes, flattened and withdrew from the cell cycle upon switching to a cell culture medium supplemented with 10% fetal bovine serum. More importantly, these tumor stem cells differentiated into Tuj1+ or NeuN+ neurons, GFAP+ astrocytes and CNPase+ oligodendrocytes, thus highlighting their multi-potency. Furthermore, these cells were capable of propagating secondary medulloblastomas when orthotopically transplanted into host mice.Download video file.^{(40M, mov)}     

泰山虫草C-2最优液体培养条件筛选

在泰山虫草资源调查过程中,采集、分离获得一株泰山虫草菌(Cordycepssp.)。对其进行人工驯化栽培,发现其生长周期短,发菌快,适应性强。利用正交试验设计法筛选其最佳液体发酵培养基及条件,用SAS数据处理软件系统对菌丝生物量、多糖进行方差分析,并测定发酵液中超氧化物歧化酶(SOD)、过氧化物酶(POD)等酶的活性。结果表明,泰山虫草最优的菌丝体发酵培养基组合是:马铃薯20%、蔗糖2%、豆粉2%、胶体几丁质0.5%、无机盐0.1%KH2PO41、2 h光照。     

抗体催化的臭氧生成及其在动脉粥样硬化发病中的潜在作用

本文采用靛蓝胭脂红褪色反应检测抗体是否能催化经佛波酯(PMA)激活的THP-1单核细胞产生臭氧(O_3),并借助荧光分光光度法检测细胞内胆固醇的积聚,以及高效液相色谱法(HPLC)分析细胞内胆固醇的臭氧氧化产物5,6-secosterol生成的情况,探讨抗体催化水的氧化作用对动脉粥样病变形成潜在的影响。结果显示,THP-1单核细胞经人IgG、PMA共同孵育后,显著产生了O_3的化学信号,对靛蓝胭脂红具有明显的褪色作用,过氧化氢酶能加强这种褪色反应,而乙烯苯甲酸具有显著的抑制作用;THP-1单核细胞经人IgG、PMA及低密度脂蛋白(LDL)共同孵育后,其细胞内积聚的总胆固醇(TC)、游离胆固醇(FC)、胆固醇酯(CE)及胆固醇酯占总胆固醇的比值均明显增多,并显著产生了胆固醇的臭氧氧化产物5,6-secosterol,乙烯苯甲酸对它们均有抑制效应。以上结果表明：激活的THP-1单核细胞在抗体的催化下具有产生臭氧的能力,可能是动脉粥样硬化形成过程中一种新的重要机制。