Progress toward re‐engineering non‐ribosomal peptide synthetase proteins: a potential new source of pharmacological agents

Many important pharmaceutical agents, including vancomycin, bleomycin, cyclosporin, and several antibiotics, are produced by non‐ribosomal peptide synthetase (NRPS) enzymes in microorganisms. The NRPS pathway produces an extensive library of products using multienzyme complexes acting in an assembly‐line fashion. Engineering an NRPS system to produce an even greater variety of products, some of which may also have beneficial therapeutic value, would be an enormous advantage. Several approaches have been successful in generating novel NRPS products: mutational biosynthesis during which nonnatural substrates are fed to an organism; domain and module swapping between different species to generate hybrid enzymes; and rational site‐directed mutagenesis, based either on phylogeny or computational prediction, intended to switch substrate specificity and produce altered products. This review will highlight the progress in these areas and describe research in the future that will extend the capacity for re‐engineering NRPS systems. Drug Dev. Res. 66:9–18, 2006. © 2006 Wiley‐Liss, Inc.     

脐动脉血PCT、CRP检测在早产儿早期感染中的作用

目的:为进一步提高早产儿重症感染的早期诊断率探讨一种快速、可靠的方法。方法:用LUMITestPCT方法检测我院出生的有感染可能的早产儿的脐动脉血清降钙素原(PCT)。同时检测脐动脉血C反应蛋白(CRP);由新生儿专业的医师对其病情进行观察,在不知道PCT值时做出病情的临床诊断,52例病例分为:重症感染组(n=28)和对照组(n=24)。进行回顾性对比分析。结果:早期开始的早产儿感染与CRP、PCT浓度在出生时的增加相关,差异有极显著性(P<0.001);脐动脉血中,PCT浓度的增加与CRP水平变化呈正相关(r=0.88)。PCT的敏感性(96.4%)和特异性(95.8%)均高于CRP(敏感性75%,特异性91.6%)。结论:PCT可作为早产儿早期感染的早期、快速检测指标,与CRP相比,PCT敏感性、特异性均高。本研究方法对早产儿无创伤。     

Spindle-cell glioblastoma or gliosarcoma?

'Gliosarcomas' have long been considered to be mixed gliomas and sarcomas. The present study failed to define criteria which clearly delineate 'gliosarcomas' from glioblastoma multiforme and suggests that 'gliosarcomas' should be considered as spindle cell glioblastomas. A total of six cases originally diagnosed as 'gliosarcomas' were compared with four cases of glioblastoma multiforme. No clinical or prognostic features were defined which would clearly separate 'gliosarcomas' from glioblastoma multiforme. Macroscopically, biopsies from 'gliosarcomas' ranged from firm, apparently well-circumscribed tumours to poorly circumscribed lesions with a soft consistency resembling glioblastoma multiforme. Histology revealed a continuous spectrum in which 'gliosarcomas' with large reticulin-rich areas of spindle cells merged with typical glioblastomas containing only small islands of spindle cells and reticulin staining. Immunocytochemistry for glial fibrillary acidic protein (GFAP); S100 protein and alpha-smooth muscle actin (ASMA) showed that the majority of cells in reticulin-poor areas of 'gliosarcoma' and glioblastomas expressed S100 protein and GFAP; many expressed ASMA and some expressed both GFAP and ASMA. Spindle cells in reticulin-rich areas of 'gliosarcomas' and glioblastomas most frequently expressed ASMA but many cells also expressed S100 protein and GFAP; some cells expressed both GFAP and ASMA. The results of this study and a review of the literature suggests that there is a clinical, radiological and pathological continuum with glioblastoma and 'gliosarcoma' at different ends of the spectrum. It is suggested, therefore, that most, if not all, 'gliosarcomas' be redesignated as spindle cell glioblastomas and not be considered as a mixture of glioma and sarcoma.     

胃癌组织p16基因蛋白表达的意义

目的　检测 p1 6基因蛋白在胃癌组织、癌旁组织中的表达及其分布特点 ,分析其与胃癌临床病理学特征及预后的关系 .方法　采用 S- P免疫组织化学法对 53例胃癌组织及 35例癌旁组织进行 p1 6蛋白的定位观察 .结果　各病理类型胃癌组织、癌旁组织均有 p1 6基因蛋白表达 .阳性率分别为 62 .3% (33/53)和 88.6% (31 /35) ,阳性细胞的棕黄色颗粒主要位于细胞核 .胃癌 p1 6基因蛋白表达与性别、年龄、肿瘤部位在统计学上无差异 (P>0 .0 5) ;而与组织学类型、病理分级、淋巴结转移、临床病理分期在统计学上有差异 (P<0 .0 5) .p1 6蛋白阳性者 5年生存率 51 .0 %高于 p1 6蛋白阴性者 2 0 .0 % (P<0 .0 5) .结论　 p1 6基因缺失和表达水平的改变与胃癌发生、发展密切相关 ,检测 p1 6基因蛋白表达可作为辅助临床判断胃癌的生物学行为及推测预后的指标     

胃癌组织中P16、Cyclin D1、Rb表达及其生物学行为的研究

目的研究P16、Cyclin、D1Rb基因在胃癌中的表达及其与胃癌生物学行为的关系.方法采用免疫组化方法检测10例正常胃黏膜、30例胃癌组织中P16、Cyclin D1、Rb蛋白的表达,并结合其临床资料进行分析.结果P16、Cyclin D1、Rb蛋白阳性表达,正常胃粘膜分别为90%、10%、90%;胃癌分别为36.67%、53.33%、50%,胃癌纽与正常对照组间差异均有显著性(P＜0.05),P16、Cyclin D1蛋白与胃癌分化程度、淋巴结转移与否、远处转移与否呈一定的相关性.结论P16、Cyclin D1、Rb基因与胃癌发生有关,P16、Cyclinp蛋白检测有助于胃癌预后的判断.     

C-反应蛋白水平对急性缺血性脑卒中预后影响的系统评价

目的 应用Meta分析,评价C-反应蛋白(C-reactive protein,CRP)水平对急性缺血性脑卒中预后的影响.方法 计算机检索PubMed、MEDLINE(1991-2006)、中国学术期刊全文数据库(1979-2006)等.搜集CRP水平对急性缺血性脑卒中预后影响非随机对照研究,逐篇评价纳入文献的质量,采用RevMan4.2版软件进行Meta分析.结果 共纳入7篇文献(外文4篇,中文3篇),涉及841例患者.在急性缺血性脑卒中患者中,3个月以上随访截尾时间的死亡率和再发血管事件与CRP水平相关,CRP增高组发生率比CRP正常组分别高2.71和1.81倍[OR合并分别为3.71和2.81,95%CI分别为(2.40,5.73)和(1.42,5.53)]均有统计学意义(P＜0.05);CRP测量值与缺血性脑卒中复发和死亡亦相关,在复发患者中,CRP测量值平均高于未复发者为3.52 mg/L[WMD=3.52,95%CI(-0.01,7.05)],具有一定的临床意义;在死亡患者中,CRP测量值平均高于生存者为8.02 mg/L[WMD=8.02,95%CI(5.68,10.37)],具有统计学意义(P＜0.05).结论 发病时CRP的水平可以初步评价急性缺血性脑卒中患者的预后.     

银杏叶提取物联合脑蛋白水解物治疗脑梗塞68例

目的观察银杏叶提取物与脑蛋白联用治疗脑梗塞的疗效及对血液流变学指标的影响。方法132例脑梗塞患者随机分为治疗组及对照组。治疗组给舒血宁20 mL,脑蛋白注射液60 mg,1次/d静滴,连用14 d观察治疗前后疗效及血流变指标变化。结果与对照组比较治疗前后疗效及血流变指标均有显著改善。结论银杏叶提取物(舒血宁)合用脑蛋白注射液治疗脑梗塞,可降低血粘度,促进神经功能改善。     

Influence of Genetic Variation in the C-Reactive Protein Gene on the Inflammatory Response During and After Acute Coronary Ischemia

The aim of this research was to assess whether common genetic variants within the C-reactive protein gene ( CRP ) are related to the degree of acute rise in plasma C-reactive protein (CRP) levels following an acute coronary syndrome (ACS). While polymorphisms within CRP are associated with basal CRP levels in healthy men and women, less is known about the relationship of such genetic variants and the degree of CRP rise during and after acute ischemia. Plasma CRP is associated with increased rates of recurrent coronary events. We evaluated seven common genetic variants within CRP and assessed their relationship to the degree of rise in CRP levels immediately following an acute coronary syndrome in 1827 European American patients. Variants in the putative promoter region, −757T > C and −286C > T > A, were associated with the highest CRP elevations after ACS. Patients with two copies of the A allele of SNP −286C > T > A had median CRP values of 76.6 mg/L, compared to 11.1 mg/L in patients with no copies of the rare variant (p-value <0.0001), post ACS. The lowest CRP values were found for patients with minor alleles of the exonic 1059G > C and the 3'untranslated region 1846G > A SNPs. For example, patients homozygous for the minor allele of 1059G > C had 71% lower median CRP values than those homozygous for the major allele [3.5 vs 12.0 mg/L, p < 0.0001]. These trends persisted in the chronic stable phase after ischemia had resolved, and after adjustment for infarct size by peak creatinine kinase levels and clinical status by Killip class. Assessment of CRP genetic variants identified patients with higher and lower CRP elevation after acute coronary syndrome.     

Immunohistochemical and immunoelectron microscopical characterization of cerebrovascular and senile plaque amyloid in aged dogs'' brains

Immunohistochemical and immunoelectron microscopical studies were carried out on 28 aged dogs' brains. Amyloid deposits were seen in the arteries and capillaries in the leptomeninges and in superficial areas of the cortices in 19 (67.9%) of the 28 dogs (10-22 years of age). Immunohistochemically, these amyloid deposits were reactive for anti-beta/A4 antibody. Additionally, a variable number of parenchymal deposits with diffuse beta/A4-immunoreactivity (diffuse plaques) was also noted throughout the cerebral cortex in 24/28 dogs (85.7%). However, these plaque lesions were undetectable in Congo red staining. Electron microscopically, amyloid fibrils, measuring 10 nm in width, were located mainly in the tunica media of the arteries, and in less involved vessels they tended to be present among collagen fibres in the adventitia and smooth muscle cells in the outer layer of the media. The plaque lesions appeared to contain sparse aggregations of amyloid fibrils. In immunoelectron microscopical examinations, all amyloid fibrils in both blood vessels and plaques were selectively labelled by gold particles. These findings indicate that aged dogs can provide a useful experimental model for research into the beta/A4-type of cerebral amyloidosis commonly seen in Alzheimer's disease.