Quantitative PCR on 5 genes reliably identifies CTCL patients with 5% to 99% circulating tumor cells with 90% accuracy

We previously identified a small number of genes using cDNA arrays that accurately diagnosed patients with Sézary Syndrome (SS), the erythrodermic and leukemic form of cutaneous T-cell lymphoma (CTCL). We now report the development of a quantitative real-time polymerase chain reaction (qRT-PCR) assay that uses expression values for just 5 of those genes: STAT4, GATA-3, PLS3, CD1D, and TRAIL. qRT-PCR data from peripheral blood mononuclear cells (PBMCs) accurately classified 88% of 17 patients with high blood tumor burden and 100% of 12 healthy controls in the training set using Fisher linear discriminant analysis (FLDA). The same 5 genes were then assayed on 56 new samples from 49 SS patients with blood tumor burdens of 5% to 99% and 69 samples from 65 new healthy controls. The average accuracy over 1000 resamplings was 90% using FLDA and 88% using support vector machine (SVM). We also tested the classifier on 14 samples from patients with CTCL with no detectable peripheral involvement and 3 patients with atopic dermatitis with severe erythroderma. The accuracy was 100% in identifying these samples as non-SS patients. These results are the first to demonstrate that gene expression profiling by quantitative PCR on a selected number of critical genes can be employed to molecularly diagnosis SS.     

Carcinogens induce genome-wide loss of heterozygosity in normal stem cells without persistent chromosomal instability

Widespread losses of heterozygosity (LOH) in human cancer have been thought to result from chromosomal instability caused by mutations affecting DNA repair/genome maintenance. However, the origin of LOH in most tumors is unknown. The present study examined the ability of carcinogenic agents to induce LOH at 53 sites throughout the genome of normal diploid mouse ES cells. Brief exposures to nontoxic levels of methylnitrosourea, diepoxybutane, mitomycin C, hydroxyurea, doxorubicin, and UV light stimulated LOH at all loci at frequencies ranging from 1-8 x 10(-3) per cell (10-123 times higher than in untreated cells). This greatly exceeds the frequencies at which these agents have been reported to induce point mutations and is comparable to the rates of LOH observed in ES cells lacking the gene responsible for Bloom syndrome, an inherited DNA repair defect that results in greatly increased risk of cancer. These results suggest that LOH contributes significantly to the carcinogenicity of a variety of mutagens and raises the possibility that genome-wide LOH observed in some human cancers may reflect prior exposure to genotoxic agents rather than a state of chromosomal instability during the carcinogenic process. Finally, as a practical matter, chemically induced LOH is expected to enhance the recovery of homozygous recessive mutants from phenotype-based genetic screens in mammalian cells.     

VEGF is up-regulated by hypoxic stimulation and related to tumour angiogenesis and severity of disease in oral squamous cell carcinoma: in vitro and in vivo studies

The present study was aimed to speculate whether the up-regulation of VEGF in oral squamous cell carcinoma (OSCC) is associated with oxygen levels, tumor angiogenesis and severity of disease. Under different oxygen levels, VEGF protein production in two oral cancer cell lines was quantitatively documented by using ELISA kit. Correlations between expression of VEGF, microvessel density, and various clinico-pathologic factors were studied in forty patients with OSCC. VEGF production was continuously elevated in supernatants from both cell lines in respond to the drop of oxygen levels. When oxygen level decreased to 1%, there was a 2.1-fold and nearly a 2.9-fold elevation of VEGF production in TSCCa and GNM cell line, respectively. On hypoxia VEGF production also presented a time-dependent up-regulation in both oral cancer cell lines. VEGF positivity was correlated with regional lymph nodal involvement and clinical stage. Microvessel density was significantly higher in VEGF-positive tumors than in VEGF-negative tumors. The presence of hypoxia in oral cancers is partly responsible for the up-regulation of VEGF. The elevation of VEGF expression in OSCC tissues correlates with the increased microvessel density and severity of the disease.     

基本药物制度的利益相关者分析

目的分析基本药物制度的各个利益相关者,使这一制度能够获得可持续的发展。方法运用利益相关者理论进行分析。结果与结论只有正确处理各利益相关者的关系,引入利益相容机制和善治原则,并充分发挥媒体的作用,才能保证基本药物制度的可持续发展。     

Anti-HIV-1 activity of lignans from the fruits of <Emphasis Type="Italic">Schisandra rubriflora</Emphasis>

This study investigated the 70% aqueous acetone extract of the fruits of Schisandra rubriflora which led to the isolation of eight lignans, including a new isolate, rubrisandrin C (1), and seven known lignans (2–8). The structure of 1 was established by extensive 1D and 2D NMR spectroscopy and its absolute stereochemistry was determined by CD spectrum. Compounds 1–5 and 7–8 were evaluated for their anti-HIV-1 activity that showed inhibitory activity on HIV-1_IIIB induced syncytium formation with EC₅₀ values in the range of 2.26∼20.4 μg/mL. Compounds 1 and 7 exerted their obvious protection of HIV-1_IIIB inducted MT-4 host cells lytic effects with a selectivity index of 15.4 and 24.6, respectively.     

Topiramate: a new agent for patients with alternating hemiplegia of childhood

Alternating hemiplegia of childhood is a rare syndrome characterized by the onset, before 18 months of age, of frequent attacks of alternating paralysis. Here we report the efficacy of topiramate in four patients with alternating hemiplegia of childhood (AHC) that did not respond to flunarizine, as well as in two newly diagnosed patients. Following treatment with topiramate, the frequency and duration of hemiplegic attacks significantly improved in all patients. Additional symptoms such as seizures, migraine, involuntary movements, autonomic symptoms, and impaired mental development also improved. Topiramate is worth trying when treating patients with AHC as a first trial, or a substitute for flunarizine once the latter agent loses effect.     

Bypass to the left coronary artery system may accelerate left main coronary artery negative remodeling and calcification

Aims

This study aimed to use intravascular ultrasound (IVUS) data to reveal the mechanism of lesion progression in the native coronary circulation proximal to bypass grafts after coronary artery bypass grafting (CABG).

Methods and results

We reviewed IVUS images in 86 patients with an angiographically significant left main coronary artery (LMCA) stenosis. Overall, 41 patients underwent CABG more than 6 months (mean 8.2 ± 6.1 years) previously and had at least one patent graft to the left coronary artery system. The number of patent grafts to the left coronary artery was 1.4 ± 0.7. Comparing patent graft vs. non-CABG groups, external elastic membrane and lumen areas and remodeling index at the minimum lumen area (MLA) site trended smaller with no difference in the plaque & media area. In addition, patients in the patent graft group had more LMCA calcium whether defined by cross-sectional (arc at the MLA site of 141 ± 109° vs. 88 ± 108°, P = 0.025) or longitudinal measurements (calcium length index, calculated as LMCA calcium length divided by total LMCA length, 0.69 ± 0.38 vs. 0.50 ± 0.42, P = 0.035).

Conclusions

Negative remodeling may be the main mechanism of lesion progression proximal to a patent bypass graft, and more calcium was found in LMCA after CABG compared with non-CABG patients.     

PI3K/Akt抑制剂对人食管癌EC109细胞的生长抑制及促凋亡作用研究

目的探讨磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)特异性抑制剂LY294002对人食管癌EC109细胞生长抑制及凋亡的影响。方法通过不同浓度的LY294002作用于人食管癌EC109细胞,MTT法检测EC109细胞抑制率,流式细胞仪检测细胞凋亡率,Hoechst荧光染色和倒置相差显微镜观察EC109细胞形态学变化。结果经LY294002作用后,EC109细胞凋亡率增加,其生长明显受到抑制,经40μmol/L的LY294002作用72h后,荧光染色和相差显微镜观察到EC109细胞呈现典型细胞凋亡改变。且LY294002对食管EC109细胞的抑制率与LY294002的浓度及作用时间呈正相关(r为0.942、0.963,P<0.01),凋亡率亦与LY294002的浓度及作用时间均呈显著正相关(r为0.975、0.938,P<0.01)。结论 LY294002可抑制食管癌EC109细胞生长,并诱导其凋亡。     

Penetration of different molecule sizes upon ultrasound combined with microbubbles in a superficial tumour model

Abstract

Ultrasound combined with microbubbles (USMB) has been extensively applied to enhance drug and gene targeting delivery. However, the accumulation and distribution of particle size in the range of 5–30?nm (nano drug) to the tumour and the effects of intratumoral vascular density on permeability have been rarely reported. This study investigated Evans blue (EB) and fluorescein isothiocyanate-labelled dextran (FITC-dextran) distribution in tumour tissue upon USMB with various molecular sizes (3.7?nm and 30.6?nm). USMB increased the penetration of molecules with sizes of 5-20?nm in the whole tumour tissue, especially on the rim. For a molecule with sizes of 30.6?nm, USMB only increased penetration around the rim of the tumour with minimal improvement in the central of tumour. USMB enhanced the permeability of tumour tissue and increased tumour cells dose exposure without affecting tumour blood perfusion or microvessel density. The current study served as the foundation of parameter preference for therapeutic USMB drug delivery.