晚期非小细胞肺癌组织与血浆EGFR基因表达差异及临床意义

[目的]探讨DNA表皮因子生长受体(epidermal growth factor receptor,EGFR)基因在晚期非小细胞肺癌(no-small cell lung cancer,NSCLC)患者组织和血浆中的表达差异及临床意义.[方法]选取2015年3～6月在本院初诊的NSCLC患者65例,提取血浆及组织标本DNA,检测DNA EGFR基因突变情况并比较.[结果]65例患者组织EGFR基因突变检出23例,突变率为35.4％,循环DNA中检出1 2例,突变率为18.5％.以组织标本检测为准,血浆DNA中EGFR基因突变检测的特异性为93.8％;灵敏度为43.5％.在组织标本检测中,44例男性EGFR基因突变率为22.7％;21例女性患者EGFR基因突变率为61.9％,两者比较有统计学差异(P＜0.05).41例吸烟患者EGFR基因突变率为17.1％(7/41);24例不吸烟患者EGFR突变率为66.7％(16/24),两者比较有统计学差异(P＜0.05).41例腺癌患者EGFR突变率为46.3％;24例非腺癌患者EGFR突变率为16.7％,两者比较有统计学差异(P＜0.05).血浆中EGFR基因突变的分布与肿瘤组织检测结果相符,在不同性别、不同吸烟情况及不同病理类型中差异有统计学意义(P＜0.05),在肿瘤分期方面无显著性差异(P＞0.05).[结论]对于晚期NSCLC患者,血浆循环DNA EGFR基因突变检测特异性高、敏感性低,可用于其补充诊断.     

磷脂转运蛋白在毕赤酵母中的高效表达

为深入了解磷脂转运蛋白的结构和功能，从中国人胎肝组织总RNA成功克隆了磷脂转运蛋白成熟肽基因，所得磷脂转运蛋白基因通过同源重组整合至毕酵母细胞染色体的氧化酶AOX1基因中，甲醇诱导后经SDS聚丙烯酰胺凝胶电泳显示诱导表达蛋白分子量约为75kDa，薄层扫描显示表达蛋白中酵母蛋白总量的28%，为大量制备重组磷脂转运蛋白奠定了基础。     

Urinary Nucleic Acid TSPAN13-to-S100A9 Ratio as a Diagnostic Marker in Prostate Cancer

The potential use of urinary nucleic acids as diagnostic markers in prostate cancer (PCa) was evaluated. Ninety-five urine samples and 234 prostate tissue samples from patients with PCa and benign prostatic hyperplasia (BPH) were analyzed. Micro-array analysis was used to identify candidate genes, which were verified by the two-gene expression ratio and validated in tissue mRNA and urinary nucleic acid cohorts. Real-time quantitative polymerase chain reaction (qPCR) was used to measure urinary nucleic acid levels and tissue mRNA expression. The TSPAN13-to-S100A9 ratio was selected to determine the diagnostic value of urinary nucleic acids in PCa (P = 0.037) and shown to be significantly higher in PCa than in BPH in the mRNA and nucleic acid cohort analyses (P < 0.001 and P = 0.013, respectively). Receiver operating characteristic (ROC) analysis showed that the area under the ROC curve was 0.898 and 0.676 in tissue mRNA cohort and urinary nucleic acid cohort, respectively. The TSPAN13-to-S100A9 ratio showed a strong potential as a diagnostic marker for PCa. The present results suggest that the analysis of urine supernatant can be used as a simple diagnostic method for PCa that can be adapted to the clinical setting in the future.     

Design of self-assembling peptide hydrogelators amenable to bacterial expression

Hydrogels formed from self-assembling peptides are finding use in tissue engineering and drug delivery applications. Given the notorious difficulties associated with producing self-assembling peptides by recombinant expression, most are typically prepared by chemical synthesis. Herein, we report the design of a family of self-assembling β-hairpin peptides amenable to efficient production using an optimized bacterial expression system. Expressing peptides, EX1, EX2 and EX3 contain identical eight-residue amphiphilic β-strands connected by varying turn sequences that are responsible for ensuring chain reversal and the proper intramolecular folding and consequent self-assembly of the peptide into a hydrogel network under physiological conditions. EX1 was initially used to establish and optimize the bacterial expression system by which all the peptides could be eventually individually expressed. Expression clones were designed to allow exploration of possible fusion partners and investigate both enzymatic and chemical cleavage as means to liberate the target peptide. A systematic analysis of possible expression systems followed by fermentation optimization lead to a system in which all three peptides could be expressed as fusions with BAD-BH3, the BH3 domain of the proapoptotic BAD (Bcl-2 Associated Death) Protein. CNBr cleavage followed by purification afforded 50, 31, and 15 mg/L yields of pure EX1, EX2 and EX3, respectively. CD spectroscopy, TEM, and rheological analysis indicate that these peptides fold and assembled into well-defined fibrils that constitute hydrogels having shear-thin/recovery properties.     

Role of miR-511 in the Regulation of OATP1B1 Expression by Free Fatty Acid

MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter. We sought to evaluate the effect of miR-511 on the regulation of OATP1B1 expression by free fatty acids. When using free fatty acids to stimulate Chang liver cells, we found that the expression of miR-511 increased significantly while the expression of OATP1B1 decreased. We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay. Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells. Our study indicates that miR-511 may play an important role in the regulation of OATP1B1 expression by free fatty acids.     

诱癌过程中鼠肝组织一氧化氮合酶的表达与改变

目的：研究ＮＯ合酶在肝癌发生过程中的表达与改变。方法：以化学致癌剂２－ＦＡＡ喂饲ＳＤ大鼠，对鼠肝细胞在癌变过程中的组织学改变、一氧化氮合酶（ＮＯＳ）的表达和动态变化进行了研究。结果：当肝细胞轻度损伤时，肝组织中ＮＯＳ活性与对照组相比明显下降；当肝细胞表现为癌前病变和肝细胞完全癌变时，肝组织中ＮＯＳ的合成增加其比活性仍低于正常对照鼠；肝组织中ＮＯＳ与ＮＯ水平呈明显正相关（ｒ＝０５７，Ｐ＜００１），而与总ＲＮＡ浓度无相关性。结论：研究资料提示ＮＯＳ在诱癌早期的表达处于抑制状态以对肝细胞起保护作用；而肝癌形成阶段，ＮＯＳ加速表达以杀伤肿瘤细胞     

Characterization of Monocyte-Macrophage-Lineage Cells Induced from CD34<Superscript>+</Superscript> Bone Marrow Cells In Vitro

We characterized the expression of cell surface antigens and cytokine-secreting ability of monocyte-macrophage-lineage cells induced in vitro from CD34+ bone marrow cells. After cultivation for 3 weeks, we observed 2 distinct cell fractions: a floating small, round cell fraction and an adherent large, protruding cell fraction. Both cell fractions expressed myelocyte-monocyte-lineage antigens, but mature-macrophage markers such as CD206 were expressed only by the adherent cells. An assessment of cells cultured for 5 weeks revealed spontaneous secretion of interleukin 8 (IL-8) and IL-6, and lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-alpha) secretion in both fractions, but only the adherent cell fraction secreted IL-10 after LPS stimulation. In contrast, both fractions of cells cultured for 3 weeks spontaneously secreted low levels of IL-8, but none of the other cytokines. Upon LPS stimulation, the cells secreted IL-6 and TNF-alpha, but not IL-10. We also assessed the effect of granulocyte colony-stimulating factor (G-CSF) pretreatment on TNF-alpha secretion by each cell fraction and found that G-CSF reduced TNF-alpha secretion only in the adherent fraction of cells cultured for 3 weeks. Monocyte-macrophage-lineage cells induced in vitro should provide an ideal model for functional analysis of monocyte-macrophage cells.     

朱震亨从脾胃病探析误诊误治

[目的]通过探析《局方发挥》之脾胃病误诊误治,以期指导与改进现代医家临床治疗。[方法]对《局方发挥》之脾胃病误诊误治进行整理分析,揭示朱震亨从脾胃病探析误诊误治的特色。[结果]朱震亨从脾胃病论误诊误治的特色:1)《和剂局方》具有误导性脾胃病治疗的中医知识,导致当时医者与患者误诊误治脾胃病。2)《和剂局方》引起的脾胃病误诊误治问题,主要可分成两类:辨证失误和治法失误。3)辨证失误所导致的误诊误治包括忽视审证求因、认为"诸寒为病"、误辨寒热虚实等。4)治法失误包括"一方通治诸病"、滥用辛香燥热药组方、利用砒丹巴硇行积垢、修改汤剂为丸剂、用峻剂盲目祛邪及错误滋补等。[结论]朱震亨在《局方发挥》指出当时脾胃病的误诊误治问题,显示出了其独特见解,对现代医家治疗脾胃病,具有一定的指导作用与参考价值。     

日本血吸虫重组抗原基因的高效表达和特性鉴定

为了获得有效的保护性抗原分子，我们对已构建的日本血吸虫成虫ｃＤＮＡ库的免疫筛选中所获得的阳性克隆λＳｊ５１４的。ＤＮＡ插入片段进行ＰＣＲ扩增，扩增产物亚克隆入高表达载体ｐＧＥＸ－１λｔ，得到高效表达克隆ｐＧＳｊ２４。经诱导表达生产出约２０ｋＤａ的分离表达产物，该特异性蛋白可被日本血吸虫免疫血清、感染兔血清和病人血清特异地识别，而且具有较强的免疫原性，可刺激动物产生较强的抗体反应。