Targeting the HGF/SF receptor c-met using a hammerhead ribozyme transgene reduces in vitro invasion and migration in prostate cancer cells

BACKGROUND: Hepatocyte growth factor scatter factor (HGF/SF) elicits a number of biological activities including invasion and migration through activation of its tyrosine kinase receptor c-Met. Over expression of c-Met has been implicated in prostate cancer development and progression. This study examined the effect of a ribozyme transgene, designed to inhibit human c-Met expression, and its impact on in vitro invasion and migration in prostate cancer. METHODS: A transgene (Met 560) consisting of U1 snRNA, hammerhead ribozyme, and antisense was cloned into a modified pZeoU1-EcoSpe vector and transfected into DU-145 cells. The effect of HGF/SF was tested on prostate cancer cells whose expression of c-Met had been blocked by way of a ribozyme transgene. RESULTS: Met 560 stable transfectants (DU-145(+/+)) manifested a complete loss of c-Met expression at mRNA and protein levels. In contrast, control plasmid (DU-145(+/-)) and wild-type DU-145 cells (DU-145(-/-)) had similar levels of c-Met expression. HGF/SF significantly increased the in vitro invasiveness (mean 47.71 +/- SE 7.75; P < 0.01 vs. control 24.14 +/- 1.34), and migration (mean 48.44 +/- SE 3.51; P < 0.01 vs. control 22.95 +/- 1.47) of DU-145(-/-) cells, respectively. Similarly, HGF/SF also increased the invasion (62.33 +/- 6.34; P < 0.001 vs. control 24.5 +/- 2.35) and migration (46.14 +/- 2.26; P < 0.01 vs. control 21.82 +/- 1.62) of DU-145(+/-) cells. In contrast, DU-145(+/+) cells had lost its response to HGF/SF induced invasion (22.33 +/- 2.08; P > 0.05 vs. control 23.5 +/- 2.11) and migration (24.12 +/- 0.86; P > 0.05 vs. control 23.27 +/- 0.81). CONCLUSIONS: Targeting the HGF/SF receptor by way of a hammerhead ribozyme encoding antisense to c-Met, is an effective method to reduce the invasive or migration potential in prostate cancer, and may have important therapeutic implications.     

Australian implantable cardiac defibrillator recipients: quality-of-life issues

Implantable cardioverter defibrillators (ICDs) have become a well-established therapy for people experiencing potentially lethal dysrhythmias. Australian recipients' quality of life and adjustment to the device over time, device-related complications, shock and associated sensations, and potential sequelae have not been widely explored. This paper reports a longitudinal prospective study of Australian ICD recipients (n = 74) to determine their responses to the device, health-related quality of life over time and shock experiences. A questionnaire designed for the study and the Medical Outcomes Trust Quality of Life Instrument, the SF36, were completed by recipients prior to and at 3 and 12 months post insertion. Results show that quality of life decreased for general health and social function between 3 and 12 months. Nearly half (49%) of the recipients received shocks within 12 months and the majority (92%) of these experienced sequelae that could make driving hazardous. Half of the population (49%) were driving at 3 months and 69% by 12 months, including 67% of those who had been shocked. Twenty-seven percent were hospitalized with device-related complications. Driving, the shock experience and rehospitalization, the shock experience and driving behaviour are significant issues for those with the implanted device. While it is a limitation of the study that partners and carers were not included, these findings will also be of interest to them.     

Isodiospyrin as a novel human DNA topoisomerase I inhibitor

Isodiospyrin is a natural product from the plant Diospyros morrisiana, which consists of an asymmetrical 1,2-binaphthoquinone chromophore. Isodiospyrin exhibits cytotoxic activity to tumor cell lines but very little is known about its cellular target and mechanism of action. Unlike the prototypic human topoisomerase I (htopo I) poison camptothecin, isodiospyrin does not induce htopo I-DNA covalent complexes. However, isodiospyrin antagonizes camptothecin-induced, htopo I-mediated DNA cleavage. Binding analysis indicated that isodiospyrin binds htopo I but not DNA. These results suggest that isodiospyrin inhibits htopo I by direct binding to htopo I, which limits htopo I access to the DNA substrate. Furthermore, isodiospyrin exhibits strong inhibitory effect on the kinase activity of htopo I toward splicing factor 2/alternate splicing factor in the absence of DNA. Thus, these findings have important implications on naphthoquinone and its derivatives' cellular mode of actions, i.e. these novel DNA topoisomerase I inhibitors can prevent both DNA relaxation and kinase activities of htopo I.     

An empirical comparison of EQ-5D and SF-6D in liver transplant patients

There remains disagreement about the preferred utility-based measure of health-related quality of life for use in constructing quality-adjusted life years (QALYs). The recent development of a new measure, the SF-6D, has highlighted this issue. The SF-6D and EuroQol EQ-5D measure health-related utilities on a scale where 0 represents death and 1 represents full health, and both have utility scores generated from random samples of the general UK population. This study explored whether, in a large sample of liver transplant patients, the two instruments provide similar results. The empirical data highlight important variation in the results generated from the use of the two instruments. The data are consistent with a view that the SF-6D does not describe health states at the lower end of the utility scale but is more sensitive than EQ-5D in detecting small changes towards the top of the scale.     

膀胱癌患者手术治疗前后血清SE-cad、SF和TSGF检测的临床意义

目的：探讨了膀胱癌患者手术治疗前后血清SE-cad、SF和TSGF水平的变化及临床意义。方法：应用放免法、生化法和酶免法对36例膀胱癌患者进行了血清SE-cad、SF和TSGF检测,并与30名正常健康人作比较。结果：在手术前血清SE-cad、SF和TSGF水平非常显著地高于正常人组（P〈0.01）,手术治疗后3个月则与正常人比较无显著性差异（P〉0.05）。结论：检测膀胱癌患者血清中SE-cad、SF和TSGF水平的变化对诊断、治疗和预后观察均具有重要的临床价值。     

SF器械在胸腰椎骨折治疗的临床应用研究

自2001～2003年,我们应用SF内固定器[1](苏州大学第一附属医院洪天禄设计,江苏康辉医疗器械厂生产)治疗胸腰椎骨折32例,取得了满意效果.现总结报告如下.     

SF-36问卷应用于老年人群生命质量的研究

目的：利用SF－36问卷调查社区老年人群的生命质量，探讨影响生命质量的因素。方法：面对面访谈调查生命质量及影响因素，利用Logistic逐步回归分析调查资料。结果：社区老年人群的生理健康评分较好，但心理健康较差。年龄、性别、婚姻状况和医疗费用是生理健康的主要影响因素；年龄、性别、经济收入和子女状况是心理健康的主要影响因素。结论：一些人口社会学特征影响老年人的生命质量，因此，家庭和社会应该更多地关注老年人的生活环境和医疗条件，提高老年人的健康水平。     

The ACMG SF v3.0 gene list increases returnable variant detection by 22% when compared with v2.0 in the ClinSeq cohort

PurposeThe American College of Medical Genetics and Genomics (ACMG) recommends the return of pathogenic and likely pathogenic (P/LP) secondary findings from exome and genome sequencing. The latest version (ACMG secondary finding [SF] v3.0) includes 14 additional genes. We interrogated the ClinSeq cohort for variants in these genes to determine the additional yield in unselected individuals.MethodsExome data from 1473 individuals (60% White, 34% African American or Black, 6% other) were analyzed. We restricted our analyses to coding variants; +1,+2,–1, and –2 splice site variants; and the pathogenic GAA variant, NM_000152.5:c.-32-13T>G. Variants were assessed with slightly modified ACMG/Association of Molecular Pathology guidelines.ResultsA total of 25 P/LP variants were identified. In total, 7 individuals had P/LP variants in genes recommended for return of heterozygous variants, namely HNF1A (1), PALB2 (3), TMEM127 (1), and TTN (2). In total, 4 individuals had a homozygous variant in a gene recommended for biallelic variant return, namely HFE, NM_000410.3(HFE):c.845G>A p.Cys282Tyr. A total of 17 P/LP variants were identified in the heterozygous state in genes recommended only for biallelic variant reporting and were not returned. The frequency of returnable P/LP variants did not significantly differ by race.ConclusionUsing the ACMG SF v3.0, the returnable P/LP variant frequency increased in the ClinSeq cohort by 22%, from 3.4% (n = 50, ACMG SF v2.0) to 4.1% (n = 61, ACMG SF v3.0).     

Functional characterization of five NR5A1 gene mutations found in patients with 46,XY disorders of sex development

Steroidogenic factor‐1 (SF1), encoded by the NR5A1 gene, is a key regulator of steroidogenesis and reproductive development. NR5A1 mutations described in 46,XY patients with disorders of sex development (DSD) can be associated with a range of conditions of phenotypes; however, the genotype–phenotype correlation remains elusive in many cases. In the present study, we describe the impact of five NR5A1 variants (three novel: p.Arg39Cys, p.Ser32Asn, and p.Lys396Argfs*34; and two previously described: p.Cys65Tyr and p.Cys247*) on protein function, identified in seven patients with 46,XY DSD. In vitro functional analyses demonstrate that NR5A1 mutations impair protein functions and result in the DSD phenotype observed in our patients. Missense mutations in the DNA binding domain and the frameshift mutation p.Lys396Argfs*34 lead to both, markedly affected transactivation assays, and loss of DNA binding, whereas the mutation p.Cys247* retained partial transactivation capacity and the ability to bind a consensus SF1 responsive element. SF1 acts in a dose‐dependent manner and regulates a cascade of genes involved in the sex determination and steroidogenesis, but in most cases reported so far, still lead to a sufficient adrenal steroidogenesis and function, just like in our cases, in which heterozygous mutations are associated to 46,XY DSD with intact adrenal steroid biosynthesis.     

MCV、RDW、LDH和SF联合检测对骨髓增生异常综合征与巨幼细胞贫血鉴别诊断的临床意义

目的探讨MCV、RDW、乳酸脱氢酶（LDH）和铁蛋白（SF）联合检测对骨髓增生异常综合征（MDS）和巨幼细胞贫血（MA）鉴别诊断的临床意义。方法选取临床确诊的29例巨幼细胞贫血和34例骨髓增生异常综合征的初诊患者,严格按照操作规程,应用德国罗氏公司生产的RocheMODU-LARPPI自动生化分析仪采用酶速率法原理测定血浆LDH,用全自动电化学发光免疫分析仪E170采用电化学发光原理检测血浆铁蛋白及应用美国雅培CD3700全自动血常规仪测定全血各项参数并自动演算出MCV和RDW数值。两组之间比较采用t检验。结果 MDS组和MA组的MCV、RDW、LDH和SF比较均存在显著性差异（P〈0.05）。结论联合检测MCV、RDW、LDH和SF对鉴别诊断MDS和MA具有临床意义,并有显著的社会效应,值得临床推广应用。