Recovery of cryopreserved 96-well plates from −80° C

Summary A simple method is described for the effective recovery of cells cryopreserved in 96-well plates.     

柯萨奇B4病毒非结构蛋白P2C重组质粒的构建

目的：构建柯萨奇B4病毒非结构蛋白P2C重组质粒。方法：用RT-PCR技术，从柯萨奇B4病毒中钓取非结构蛋白P2C cDNA片段，克隆入PUCan-T载体，转化大肠杆菌DH5α，构建重组质粒。结果：以柯萨奇B4病毒的总RNA为模板，扩增出987bp大小的片段，克隆入PUCan-T载体，用BamHⅠ、HindⅢ双酶切鉴定，与非结构蛋白P2C的PCR扩增产物片段大小一致。结论：成功地构建了柯萨奇B4病毒非结构蛋白P2C重组质粒。     

The Distribution of Synaptotagmin II in RBL-2H3 and Its Regulation on Exocytosis of Lysosomes in RBL-2H3

Synaptotagmin(Syt)constitutes a family of membrane-trafficking proteins,so far nearly 20 Syts have beendiscovered.Extensive work showed that synatotagmins were a potential Ca~(2+) sensor for regulated exocytosis.Thisstudy was to investigate the expression and location of synaptotagmin Ⅱ(Syt2)in RBL-2H3(RBL)and its role inregulating exocytosis of RBL.The expression of Syt2 in RBL was confirmed by Western blot.The recombinantexpression vector pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation,the stabletransfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated atplasma membrane with little detected in cytoplasm.To analyze the role of Syt2 during exocytosis of RBL,therelease of cathepsin D was assayed by immunoblotting.Compared with control,the release of cathepsin D byRBL-Syt2-S was markedly decreased.The results indicated that Syt2 played a negative regulation in exocytosis oflysosomes in RBL.Cellular & Molecular Immunology.2005;2(3):205-209.     

基于小波分解的多尺度医学图像融合技术

给出了一种基于小波分解的多尺度图像融合新方法。其基本思想是 ,先对源图像进行小波多尺度分解 ;其次 ,采用了基于区域特性量测选择的加权算子的融合规则进行小波系数融合 ;最后通过小波逆变换重构融合图像。实验结果表明 ,该融合方法十分有效 ,融合图像完好地显示了源图像各自的信息。     

蛋白转导域介导BCR/ABL抗原对CML患者T细胞的活化作用

目的：研究蛋白转导域(PTD)介导的BCR／ABL抗原对慢性髓细胞白血病(CML)患者T细胞的特异性活化作用。方法：利用基因工程技术，将PTD基因与CML b3a2 bcr／abl基因融合并原核表达。将纯化的PTD—BCR／ABL融合蛋白与CML患者外周血单个核细胞(PBMC)体外共孵育，用流式细胞仪分别检测CD4^ 、CD8^ T细胞上活化抗原CD25的表达。结果：终浓度为100mg／L的PTD—BCR／ABL抗原体外刺激4d后，10例CML患者中，5例表现为CD8^ T细胞活化，2例表现为CD4^ T细胞活化，其中有1例CD8^ 和CD4^ T细胞同时活化；而作为对照的BCR／ABL抗原刺激组无一例表现为CD8^ 或CD4^ T细胞活化。结论：PTD能将外源性BCR／ABL抗原转导入抗原呈递细胞内，加工呈递后激活抗原特异性CD8^ 及CD4^ T细胞，为CML特异性CD8^ 、CD4^ T细胞的体外活化及细胞免疫治疗开辟一条新的途径。     

Nasal mucosal gene expression in patients with allergic rhinitis with and without nasal polyps

BACKGROUND: Nasal polyps are a common problem that is difficult to diagnose and treat, in part because the cause of nasal polyposis is unknown. Although information on the pathogenesis of polyposis is lacking, there are reports suggesting that a genetic predisposition underlies this disorder. OBJECTIVE: We sought to better understand the basis of nasal polyposis associated with allergic rhinitis. We hypothesize that the expression of unique genes is associated with the nasal polyposis phenotype. METHODS: We examined 12000 human genes transcribed in the nasal mucosa of patients with allergic rhinitis with and without nasal polyps. Biopsy specimens of the mucosa of patients with and without polyps were obtained after the patients refrained from the use of topical or systemic steroid therapy for 2 weeks. RESULTS: Thirty-four genes were differentially expressed between the patient groups, including those for inflammatory molecules and putative growth factors. The greatest differential expression identified by the array analysis was for a group of genes associated with neoplasia, including mammaglobin, a gene transcribed 12-fold higher in patients with polyps compared with control patients with rhinitis alone. Quantitative RT-PCR confirmed this differential expression and documented that the number of mammaglobin mRNA copies is actually 64-fold greater in tissues of patients with polyps versus control patients. The specificity of mammaglobin protein expression was evaluated by means of immunohistochemistry, which showed specific staining in nasal polyp mucosal goblet cells only in patients with polyps. CONCLUSION: These data suggest that nasal polyposis involves deregulated cell growth, using gene activation in some ways similar to a neoplasm. In addition, mammaglobin, a gene of unknown function associated with breast neoplasia, might be related to polyp growth.     

Fusion of vestibular and podokinesthetic information during self-turning towards instructed targets

When observers step about their vertical axis ("active turning") without vision they dispose of essentially two sources of information that can tell them by how much they have turned: the vestibular cue which reflects head rotation in space and the "podokinesthetic" cue, a compound of leg proprioceptive afferents and efference copy signals which reflects the observer's motion relative to his support. We ask how these two cues are fused in the process leading to the perception of self-displacement during active turning. To this end we compared the performance of observers in three angular navigation tasks which differed with regard to the number and type of available motion cues: (1) Passive rotation, vestibular cue ( ves) only; observers are standing on a platform which is being rotated. (2) Treadmill stepping, podokinesthetic cue ( pod) only; observers step counter to the rotating platform so as to remain stable in space. (3) Active turning, ves and pod available; observers step around on the stationary platform. In all three tasks, angular velocity varied from trial to trial (15, 30, 60 degrees /s) but was constant during trials. Perception was probed by having the observers signal when they thought to have reached a previously instructed angular displacement, either in space or relative to the platform ("target"; range 60-1080 degrees ). Performance was quantified in terms of the targeting gain (displacement reached by the observer divided by target angle) and of the random error ( E(r)), which records an observer's deviation during single trials from his average performance. Confirming previous observations, E(r) was found to be significantly smaller during active turning than during passive turning, and we now complement these observations by showing that it is also significantly smaller than during treadmill stepping. This behaviour of E(r) is compatible with the idea that ves and pod be averaged during active turning. On the other hand, the observed characteristics of the targeting gain ( G(T)) support this idea only for the case of fast rotations (60 degrees /s); at lower velocities, the gain found during active turning was clearly not the average of the G(T) values recorded in the passive and the treadmill modes. We therefore also discuss alternative scenarios as to how ves and pod could interact, among these one based on the concept of a vestibular eigenmodel. A common denominator of these scenarios is that ves assumes the role of a prerequisite for an optimal use of pod during turning on a stationary support, without itself entering the calculation of displacement perception; this perception would be based exclusively on pod. Finally, it was a consistent observation that during passive rotations cognitive mechanisms fill in for the decaying vestibular signal in the context of the present navigation task, enabling observers to achieve large displacements surprisingly well although the duration of these movements exceeds by far the conventionally cited value of the central vestibular time constant (=20 s).