绿色荧光蛋白转基因胎鼠神经干细胞的超顺磁性氧化铁标记

目的探讨超顺磁性氧化铁（SPIO）标记对绿色荧光蛋白（GFP）转基因胎鼠神经干细胞（NSCs）的标记效果及标记后对其生物学特性的影响.方法采用多聚赖氨酸（PLL）介导SPIO的方法标记NSCs,用普鲁士蓝染色观察标记后NSCs内铁,比较标记和未标记NSCs的GFP表达、活力、增殖、凋亡及多向分化能力.结果普鲁士蓝染色示标记NSCS胞质内见大量蓝色颗粒,细胞的活力、增殖、凋亡及多向分化能力检测均显示两组细胞间无明显差别.结论用PLL介导SPIO标记NSCs是一种可行、安全、有效的细胞内标记方法.     

Retinal ganglion cell line apoptosis induced by hydrostatic pressure

Cellular responses to changes in pressure are implicated in numerous disease processes. In glaucoma apoptosis of retinal ganglion cells (RGCs) is associated with elevated intra-ocular pressure, however, the exact cellular mechanisms remain unclear. We have previously shown that pressure can induce apoptosis in B35 and PC12 neuronal cell lines, using an in vitro model for pressure elevation. A novel RGC line allows us to study the effects of pressure on retinal neurons. 'RGC-5' cultures were subjected to elevated ambient hydrostatic pressure conditions in our model. Experimental pressure conditions were 100 mm Hg and 30 mm Hg, representing acute (high) and chronic (lower-pressure) glaucoma, and 15 mm Hg for normal intra-ocular pressure, set above atmospheric pressure for 2 h. Negative controls were treated identically except for the application of pressure, while positive controls were generated by treatment with a known apoptotic stimulus. Apoptosis was determined by a combination of cell morphology and specific TUNEL and Annexin V fluorescent markers. These were assessed simultaneously by laser scanning cytometry (LSC), which also enabled quantitative marker analysis. RGC-5 neurons showed a significantly increased proportion of apoptotic cells compared with controls; maximal at 100 mm Hg, moderate at 30 mm Hg and not statistically significant at 15 mm Hg. This graded response, proportionate to the level of pressure elevation, is representative of the severity of analogous clinical settings (acute, chronic glaucoma and normal). These results complement earlier findings of pressure-induced apoptosis in other neuronal cultures. They suggest the possibility of novel mechanisms of pressure-related mechanotransduction and cell death, relevant to the pathogenesis of diseases such as glaucoma.     

Sertoli cell proliferation in the fetal and neonatal rat testis: a continuous phenomenon?

Sertoli cell population kinetics, as evidenced by semi-quantitative immunolabeling for proliferating cell nuclear antigen (PCNA) and Ki-67, in developing Wistar rat male gonads of embryos and neonates [14.5 days post conception (dpc)-7 days post partum (dpp)], was investigated. Throughout the examined period a gradual increase of immunolabeled Sertoli cell number, associated with intense mitotic activity, was observed. PCNA labeling index of Sertoli cells increased from 66.67 (at 14.5 dpc) to 89.74 (at 18.5 dpc) and then dropped to 75.24 (at 20.5 dpc). At birth, the percentage of PCNA immunoreactive Sertoli cells reached 98.70% and remained high thereafter, attaining a peak value of 99.90% at 7 dpp. The percentage of Ki-67 immunoreactive Sertoli cells in the fetal testis increased from E14.5 (43.95%) to E20.5 (77.40%). The proliferation rate did not alter considerably in the neonatal testis until 5 dpp. At this point, a significant increase of the Ki-67 labeling index was observed and a peak value of 95.76% was reached at 7 dpp. The pattern of Sertoli cell proliferation with age and the establishment of the final Sertoli cell number in vivo established in the present study was compared to the results from earlier investigations reported in the literature and the observed fluctuation of dividing cell numbers, associated with immunolabeling results throughout the examined period, complements and extends existing data. An appraisal of the timing of Sertoli cell proliferation in other species, namely mouse and man, is presented. The current investigation may be useful in evaluating the potential influence of factors interfering with normal mitotic activity of Sertoli cells, including cell selection mechanisms, such as apoptosis, senescence, DNA repair and hormonal/paracrine growth modulation.     

染色结肠镜对大肠隆起性黏膜病变的应用价值

目的：探讨普通结肠镜、染色结肠镜在大肠隆起性黏膜病变中临床应用价值。方法：对90例结肠镜检查发现的黏膜隆起性病变,分别行普通结肠镜、染色结肠镜诊断。所有病变均作病理学检查。比较普通结肠镜、染色结肠镜诊断结果与病理学诊断结果。结果：共发现192个隆起性黏膜病变,普通结肠镜诊断为炎性息肉、管状腺瘤、绒毛状腺瘤、大肠癌与病理诊断的符合率分别为68.8%、76.9%、66.7%和80.0%,总病理符合率为73.4%;染色结肠镜分别为86.7%、87.5%、85.7%和90.9%,总病理符合率为87.5%。结论：染色结肠镜能有效提高大肠隆起性黏膜瘤性与非瘤性病变的鉴别能力,具有较高的临床应用价值。     

FDA perspectives on health claims for food labels

The U.S. Food and Drug Administration's regulatory authority over health claims was clarified in 1990 legislation known as the Nutrition Labeling and Education Act (NLEA). This law established mandatory nutrition labeling for most foods and placed restrictions on the use of food label claims characterizing the levels or health benefits of nutrients in foods. NLEA set a high threshold for the scientific standard under which the U.S. Food and Drug Administration (FDA) may authorize health claims, this standard is known as the significant scientific agreement (SSA) standard. Subsequent legislation known as the Food and Drug Administration Modernization Act (FDAMA) provided an alternative to FDA review of the health claim where an U.S. government scientific body other than FDA concluded that there is SSA for a substance/disease relationship. Courts have since extended the scope of health claims to include qualified health claims (QHC) that are health claims not substantiated on evidence that meets the level of SSA standard, but include a qualifying statement intended to convey to the consumer the level of evidence for the claim. FDA has responded by developing an evidence-based ranking system for scientific data to determine the level of evidence substantiating a health claim. The following is an overview of FDA's regulations and evidence-based method for evaluating health claims.     

3D动脉自旋标记成像在神经胶质瘤术前分级中的应用价值

目的评估基于FSE的3D-动脉自旋标记(3D-ASL)在神经胶质瘤术前分级中的应用价值。方法选择35例疑似脑胶质瘤的患者为研究对象,术前均行MR常规、3D-ASL及MR脑灌注-动态磁敏感对比成像(DSC)。分别测得DSC和3D-ASL两种方法中的肿瘤实质部分最大相对血流量(r TBFmax)。将这两种方法得到的r TBFmax做差异分析(t检验);用两独立样本秩和检验分别比较ASL及DSC产生的r TBFmax在不同级别胶质瘤之间有无统计学差异。结果 35例患者中病理证实的脑胶质瘤患者高级别胶质瘤18例(WHOⅣ级11例,Ⅲ级7例),低级别胶质瘤13例(WHOⅡ级)。r TBFmax值在3D-ASL和DSC两种技术之间的差异无明显统计学意义(P0.05)。3 D-A S L法测得的r T B Fm a x值,高、低级别胶质瘤分别为(2.3 4±1.2 1)、(0.69±0.36),DSC法测得的r TBFmax值分别为(2.55±1.06)、(0.72±0.52),差异均有统计学意义(P0.01)。结论 3DASL可成为胶质瘤血流动力学的常规评估方法,将其与常规MRI扫描联合用于术前胶质瘤级别的评估会提高脑胶质瘤分级诊断的准确性。     

氧化型低密度脂蛋白诱导大鼠血管平滑肌细胞凋亡

为研究氧化型低密度脂蛋白引起血管平滑肌细胞凋亡及其凋亡的机制,采用细胞形态学ＤＮＡ末标记法,流式细胞仪,Ｗｅｓｔｅｎ ｂｌｔｔｉｎｇ观察氧化型低密度脂蛋白诱导培养的大鼠血管平滑肌细胞凋亡。结果发现,在氧化型低密度脂蛋白作用下,血管平滑肌细胞阻滞在分裂期的中期,并发生凋亡,ＤＮＡ末端标记法和流式细胞仪检测发现氧化型低密度脂蛋白引起细胞凋亡明显高于天然低密度脂蛋白（前者是后者的１０倍）。电镜下凋亡细胞     

The QUASAR reproducibility study, Part II: Results from a multi-center Arterial Spin Labeling test–retest study

Arterial Spin Labeling (ASL) is a method to measure perfusion using magnetically labeled blood water as an endogenous tracer. Being fully non-invasive, this technique is attractive for longitudinal studies of cerebral blood flow in healthy and diseased individuals, or as a surrogate marker of metabolism. So far, ASL has been restricted mostly to specialist centers due to a generally low SNR of the method and potential issues with user-dependent analysis needed to obtain quantitative measurement of cerebral blood flow (CBF).Here, we evaluated a particular implementation of ASL (called Quantitative STAR labeling of Arterial Regions or QUASAR), a method providing user independent quantification of CBF in a large test–retest study across sites from around the world, dubbed “The QUASAR reproducibility study”. Altogether, 28 sites located in Asia, Europe and North America participated and a total of 284 healthy volunteers were scanned. Minimal operator dependence was assured by using an automatic planning tool and its accuracy and potential usefulness in multi-center trials was evaluated as well.Accurate repositioning between sessions was achieved with the automatic planning tool showing mean displacements of 1.87 ± 0.95 mm and rotations of 1.56 ± 0.66°. Mean gray matter CBF was 47.4 ± 7.5 [ml/100 g/min] with a between-subject standard variation SD_b = 5.5 [ml/100 g/min] and a within-subject standard deviation SD_w = 4.7 [ml/100 g/min]. The corresponding repeatability was 13.0 [ml/100 g/min] and was found to be within the range of previous studies.     

Predictive Value of Measuring p53 Labeling Index at the Invasive Front of Oral Squamous Cell Carcinomas

Many studies have revealed the frequency of p53 abnormalities in oral cancer. However, it reports only on the relation between clinicopathological findings and p53 expression, and there is no study to examine the relation to the p53 labeling index (p53-LI). The purposes of this study were to examine the correlation between p53 labeling index (p53-LI) at the invasive front of oral squamous cell carcinomas (OSCC) and clinicopathological findings by immunohistochemical staining, and to evaluate clinical significance of measuring p53-LI at the invasive front of OSCC. Sixty-six biopsy specimens of OSCC were randomly selected. Patient age, gender, primary sites, T category, N category, degree of differentiation and mode of cancer invasion were analyzed. p53 expression did not correlate significantly with the clinical findings. However, significant differences were found between p53-LI and the degree of cell differentiation (p < 0.05). The p53-LI of high-grade invasive tumors was significantly larger than that of low-grade invasive tumors (p < 0.05). The overall survival rate (OS) among low-scoring p53-LI cases was 75.5% whereas that for high-scoring p53-LI cases was 40.6%. The disease-free survival rate (DFS) among low-scoring p53-LI cases was 39.5% whereas that for high-scoring p53-LI cases was 76.1%. Patients with low-scoring p53-LI had a significantly worse prognosis than those with among high-scoring p53-LI (p < 0.05). Consequently, the measurement of p53-LI at the invasive front of OSCC is significant as one of the indicators of prognosis.     

A fast,effective filtering method for improving clinical pulsed arterial spin labeling MRI

Purpose

To evaluate the effectiveness of a fully automated postprocessing filter algorithm in pulsed arterial spin labeling (PASL) MRI perfusion images in a large clinical population.

Materials and Methods

A mean and standard deviation‐based filter was implemented to remove outliers in the set of perfusion‐weighted images (control – label) before being averaged and scaled to quantitative cerebral blood flow (CBF) maps. Filtered and unfiltered CBF maps from 200 randomly selected clinical cases were assessed by four blinded raters to evaluate the effectiveness of the filter.

Results

The filter salvaged many studies deemed uninterpretable as a result of motion artifacts, transient gradient, and/or radiofrequency instabilities, and unexpected disruption of data acquisition by the technologist to communicate with the patient. The filtered CBF maps contained significantly (P < 0.05) fewer artifacts and were more interpretable than unfiltered CBF maps as determined by one‐tail paired t‐test.

Conclusion

Variations in MR perfusion signal related to patient motion, system instability, or disruption of the steady state can introduce artifacts in the CBF maps that can be significantly reduced by postprocessing filtering. Diagnostic quality of the clinical perfusion images can be improved by performing selective averaging without a significant loss in perfusion signal‐to‐noise ratio. J. Magn. Reson. Imaging 2009;29:1134–1139. © 2009 Wiley‐Liss, Inc.