采用蛋白组学技术分析质粒介导siRNA的“Off-target”效应

siRNA的"脱靶效应"(off-target effects)是RNA干扰应用研究领域的热点问题.采用蛋白组学技术对质粒介导的siRNA稳定沉默原癌基因c-myc可能存在的"off-target"效应进行初步研究,为siRNA靶向特异性的系统评价奠定理论与实验基础.构建靶向c-myc的siRNA真核表达质粒p-Mat01-1及其错配质粒p-Mis09-1,空质粒pEGFP-C1为对照,并稳定转染MCF-7人乳腺癌细胞.通过RT-PCR和Western印迹分析结果显示p-Mat01-1稳定转染克隆中c-myc/c-MYC的表达降低.采用2-DE及LC-ESI-MS/MS等方法,研究了p-Mat01-1与pEGFP-C1稳定转染克隆的蛋白组表达差异.结果显示,p-Mat01-1稳定转染克隆中有47个c-myc非调控蛋白点表达升高或降低,约占423个随机检测蛋白点的11.1%.这些蛋白涉及细胞骨架、代谢、增殖、信号传导、分子伴侣、氧化还原等多条途径.实验结果表明,质粒介导靶向c-myc的siRNA在MCF-7细胞中存在明显的"off-target"效应,提示在设计siRNA实验及应用研究时应系统考察其靶向特异性.     

Addressing long-term physical healthcare needs in a forensic mental health inpatient population using the UK primary care Quality and Outcomes Framework (QOF): an audit

Objectives This audit aims to evaluate the effectiveness of delivering an equivalent primary care service to a long-term forensic psychiatric inpatient population, using the UK primary care national Quality and Outcomes Framework (QOF).Method The audit compares the targets met by the general practitioner with special interest (GPwSI) service, using local and national QOF benchmarks (2005-2006), and determines the prevalence of chronic disease in a long-term inpatient forensic psychiatry population.Results The audit results show that the UK national QOF is a useful tool for assessment and evaluation of physical healthcare needs in a non-community based population. It shows an increased prevalence of all QOF-assessed long-term physical conditions when compared to the local East London population and national UK population, confirming previously reported elevated levels of physical healthcare need in psychiatric populations.Conclusions This audit shows that the UK General Practice QOF can be used as a standardised instrument for commissioning and monitoring the delivery of physical health services to in-patient psychiatric populations, and for the evaluation of the effectiveness of clinical interventions in long-term physical conditions. The audit also demonstrates the effectiveness of using a GPwSI in healthcare delivery in non-community based settings. We suggest that the findings may be generalisable to other long-term inpatient psychiatric and prison populations in order to further the objective of delivering an equivalent primary care service to all populations.The QOF is a set of national primary care audit standards and is freely available on the British Medical Association website or the UK Department of Health website. We suggest that primary care workers in health economies who have not yet developed their own national primary care standards can access and adapt these standards in order to improve the clinical standards of care given to the primary care populations that they serve.