Treatment with human metalloproteinase-8 gene delivery ameliorates experimental rat liver cirrhosis

BACKGROUND & AIMS: An extrahepatic human neutrophil collagenase complementary DNA (matrix metalloprotease-8) cloned in an adenovirus vector was used as a therapeutic agent in cirrhosis. METHODS: A high titer of clinical-grade AdMMP8 was obtained. RESULTS: HeLa cells transduced with AdMMP8 expressed recombinant matrix metalloprotease-8 messenger RNA and matrix metalloprotease-8 protein. Matrix metalloprotease-8 in culture sups showed enzymatic activity against native collagen type I, which was inhibited by ethylenediaminetetraacetic acid, 1,10-phenanthroline, and tissue inhibitor of metalloprotease-1. In vivo transduction showed matrix metalloprotease-8 activity, and studies to establish the efficacy of this characterized vector were performed in CCl(4) and bile duct-ligated cirrhotic rats. Transduction with 3 x 10(11) viral particles per kilogram resulted in hepatic detection of both messenger RNA and protein matrix metalloprotease-8. A consistent response in fibrosis reversal was observed in CCl(4) rats. Liver fibrosis in bile duct-ligated cirrhotic animals was decreased in 45%, along with diminished hydroxyproline content, after AdMMP8 treatment. The expression of matrix metalloprotease-2 and matrix metalloprotease-3 was up-regulated in AdMMP8 rats. Free tissue inhibitor of metalloprotease-1, as an indirect measurement of active uncomplexed matrix metalloproteases, was also increased in the AdMMP8 groups. Transforming growth factor-beta messenger RNA was diminished, and matrix metalloprotease-9 and hepatocyte growth factor increased. Treatment in both models correlated with improvements in ascites, functional hepatic tests, and gastric varices, indicating diminished intrahepatic blood pressure in animals injected with AdMMP8. CONCLUSIONS: Therefore, therapy with the matrix metalloprotease-8 gene is promising for use in a clinical setting.     

Gliomatosis cerebri evaluated by <Superscript>18</Superscript>Fα-methyl tyrosine positron-emission tomography

Gliomatosis cerebri is a rare condition in which an infiltrative glial neoplasm spreads through the brain with preservation of the underlying structure. CT and MRI show diffuse abnormal density or signal, without mass effect, and because these findings are nonspecific, it is difficult to make a definitive diagnosis. Our purpose was to assess the usefulness of a new tumour-detecting amino acid tracer for positron-emission tomography (PET), L-[3-¹⁸F] bp21u/xxlarge945.gif" alt="agr" align="BASELINE" BORDER="0">-methyl tyrosine (FMT), in patients with gliomatosis cerebri. We performed FMT PET, fluorodeoxyglucose FDG PET and MRI eight patients with gliomatosis cerebri and six with non-neoplastic disease, whose MRI also showed diffuse high signal on T2-weighted images. Standardised uptake (SUV) of FMT and FDG in the area of gliomatosis was obtained and the tumour-to-normal cortex (T/N) ratio of this was compared. The tumours were shown on FMT PET as areas of increased uptake, except in one patient with severe intracranial hypertension. There were significant differences between the SUV of FMT and the T/N ratio of FMT in patients and in controls (both P<0.01), and between the T/N ratio of FMT and FDG in patients (P <0.01). Increased uptake of FMT PET strongly suggests neoplasia. FMT PET is valuable for differentiating gliomatosis cerebri from non-neoplastic diseases showing similar diffuse high signal on T2-weighted images and little contrast enhancement.     

实时定量PCR分析健康人外周血线粒体DNA 4 977 bp缺失

目的：用实时定量PCR方法分析健康人外周血线粒体DNA4 977 bp的缺失情况。方法：收集27例年龄在17～44岁的健康人外周血,用荧光实时定量PCR方法对每个样品同时检测线粒体DNA 4 977 bp缺失的量和无4 977 bp缺失的量,进行定量分析。结果：在27例健康人外周血样品中,线粒体DNA4 977 bp缺失的阳性率为70.37%（19/27）;19例有线粒体DNA4 977 bp缺失的样品,其扩增产物的CT值在第35个循环到第39个循环之间;无线粒体DNA 4 977 bp缺失的扩增产物的CT值在第15个循环到第21个循环之间;不同年龄组间线粒体DNA 4 977 bp缺失和无线粒体DNA4 977 bp缺失的量均无统计学差异。结论：本文中超过2/3的健康人外周血含有线粒体DNA 4 977 bp的缺失;在有该缺失的样品中大约每10^6～10^7拷贝中携带有一个线粒体DNA4 977 bp缺失;随年龄的增长外周血中线粒体DNA 4 977 bp的缺失未显示出累积现象。     

DNA interactions of new antitumor platinum complexes with trans geometry activated by a 2-metylbutylamine or sec-butylamine ligand

The global modification of mammalian and plasmid DNAs by novel platinum compounds, trans-[PtCl(2)(NH(3))(Am)], where Am=2 -methylbutylamine or sec-butylamine was investigated in cell-free media using various biochemical and biophysical methods. These modifications were analyzed in the context of the activity of these new compounds in several tumor cell lines including those resistant to antitumor cis-diamminedichloroplatinum(II) (cisplatin). The results showed that the replacement of one amine group by 2-methylbutylamine or sec-butylamine ligand in clinically ineffective trans-diamminedichloroplatinum(II) (transplatin) resulted in a radical enhancement of its activity in tumor cell lines so that they are more cytotoxic than cisplatin and exhibited significant antitumor activity including activity in cisplatin-resistant tumor cells. Importantly, this replacement also markedly altered DNA binding mode of transplatin and reduced the efficiency of repair systems to remove the adducts of the new analogues from DNA. The results support the view that one strategy to activate trans geometry in bifunctional platinum(II) compounds including circumvention of resistance to cisplatin may consist in a chemical modification of the ineffective transplatin which results in an increased efficiency to form DNA interstrand cross-links.     

Role of Coagulation Factor Ⅶ in Pathogenesis of Ischemic Heart Disease

Ischemic heart disease (IHD) is a serious condition and its incidence and mortality are high and increase steadily over the past years. In recent years, researches showed that the activation of extrinsic coagulation path-way, which mainly involves FⅦ, might play an impor-tant role in thrombokinesis, and FⅦ seemed to be an independent risk factor for IHD[1]. This study was de-signed to explore the role of FⅦ activation in throm-bogenesis of IHD and provide information for early di-agnosi…     

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目的探讨儿童孤独症(CA)与多巴胺转运体(DAT1)440bp等位基因的关系。方法陕西省纺织医院等于2004年3~8月,采用PCR技术对来自西安市两所康复中心的汉族CA儿童与DAT1基因多态性进行遗传关联分析。结果(1)DAT1基因多态性中共观察到5种等位基因(320bp,360bp,440bp,480bp,520bp),6种基因型(480/480,480/320,520/480,480/360,480/440,440/440)。(2)使用相对危险度RR对CA与DAT1基因多态性的等位基因和基因型进行关联分析,显示480/440基因型和等位基因440与CA呈正关联,相对危险度分别为2·65和2·30,480/480基因型和等位基因480与CA呈负关联,相对危险度分别为0·64和0·77。但统计结果没有发现具有统计意义的差异。结论中国汉族CA儿童与DAT1440bp等位基因无遗传关联。也许等位基因440bp与基因型480bp/440bp这两个因素是发病的风险因素,还有待今后的进一步研究。     

New MspI polymorphism at +83 bp of the human apolipoprotein al gene: Association with increased circulating high density lipoprotein cholesterol levels

We recently showed that loss of a MspI restriction site in the 5′-end (intron 1) of the apolipoprotein (apo) AI gene is due to a C to T transition (+83 bp) and/or a G to A transition (+84 bp). Since this region may be relevant to the regulation of apo AI gene expression and therefore to plasma high density lipoprotein cholesterol (HDL-C), we explored the association between this MspI polymorphic site and circulating HDL-C levels in 243 healthy Caucasians. There were 143 aged 18–67 years (60 males and 83 females) and 100 aged 6–12 years (58 males and 42 females). We also compared this association with a known MspI polymorphic site, a G to A transition at −75 bp of the apo AI gene. The rare allele (−) frequency for the polymorphism at +83 bp was 4.1% and 22.1% for the polymorphism at −75 bp. Subjects heterozygous for the loss of the MspI restriction site at +83 bp (genotype: M2+−, n = 20) had higher HDL-C levels than M2++ subjects (mean ± SD: 1.73 ± 0.31 vs. 1.41 ± 0.39 mmol/l, P < 0.05 for adults; 1.71 ± 0.33 vs. 1.34 ± 0.29 mmol/l, P < 0.01 for children). Adults with the G to A substitution at −75 bp also had higher HDL-C levels (1.56 ± 0.36 mmol/l for AA, 1.53 ± 0.38 mmol/l for GA, and 1.36 ± 0.38 mmol/l for GG, P < 0.05); this difference was not observed in the children. The MspI polymorphisms at both sites were in linkage disequilibrium. Their joint effect on the HDL-C levels was also significant and individuals with rare alleles (−) at both sites had the highest HDL-C levels. In an analysis of variance, the MspI polymorphism at +83 bp, and at −75 bp and gender independently accounted for 6.5%, 1.7%, and 5.9%, respectively, of the variance in circulating HDL-C levels when age was controlled as a covariate. We conclude that loss of the MspI site by the C to T (+83 bp) and/or the G to A (+84 bp) transitions is highly associated with increased HDL-C levels. The association appears to be more significant than that of the G to A transition at −75 bp. © 1996 Wiley-Liss, Inc.     

Genetic variation in Transaldolase 1 and risk of squamous cell carcinoma of the head and neck

Background: The Pentose Phosphate Pathway (PPP) is involved in the body's protection against oxidative stress and resistance/susceptibility to apoptosis and thus has been implicated in tumor development and progression. Here we present data examining the association of genetic variation in one of the key enzymes of the PPP, Transaldolase 1 (TALDO1) with squamous cell carcinoma of the head and neck (SCCHN). Methods: We performed sequencing analysis to identify common genetic variations in TALDO1 and then investigated their association with SCCHN using samples from a population-based case/control study with both European American (EA) and African American (AA) former and current smokers. Results: We identified three polymorphisms in TALDO1 that were associated with SCCHN risk in our EA study population. Specifically the 5′ upstream variant −490C > G or T (rs10794338), which we identified as tri-allelic, showed a reduced risk compared with any presence of the common allele, odds ratio (OR) [95% confidence interval (95% CI)]: 0.57 (0.38-0.86). Additionally two intronic high frequency polymorphisms demonstrated a positive association with disease, with the presence of the variant IVS1 + 1874T > A (rs3901233), 1.76 (1.19-2.61) and IVS4 + 2187A > C (rs4963163), 1.71 (1.16-2.53). Conclusion: These results provide preliminary evidence that genetic polymorphisms in TALDO1 are associated with SCCHN.     

The p53 Arg72Pro and Ins16bp polymorphisms and their haplotypes are not associated with breast cancer risk in BRCA-mutation negative familial cases

Background: Germline disease-causing mutations in BRCA1 and BRCA2 genes confer high risk of breast and ovarian cancer, but account approximately for only 15% of familial cases. Theoretical models and experimental observations have indicated that the remaining familial aggregations would be explained by low-penetrance alleles. Moreover, alleles acting as genetic modifiers would modulate the breast cancer risk in carriers of BRCA mutations. The Ins16bp and Arg72Pro polymorphisms of p53 were implicated in breast cancer and recently it has been shown that these polymorphisms could have an effect when combined as specific haplotypes. Here, we investigated the possible role of the Ins16bp and Arg72Pro polymorphisms and their haplotypes as low-penetrance alleles in familial breast cancer. Methods: The Ins16bp and Arg72Pro polymorphisms were genotyped in a total of 350 familial index cases affected with breast cancer and negative for mutations in BRCA genes, and 352 controls. The Ins16bp and Arg72Pro polymorphisms were studied separately, and as haplotypes and haplotypes combinations. Results: None of the performed analyses resulted statistically significant. Conclusions: These observations suggested that neither the Ins16bp or Arg72Pro polymorphisms considered separately, nor any related haplotype, were associated with breast cancer risk in BRCA-mutation negative familial cases.