RNA versus DNA (NucliSENS EasyQ HIV-1 v1.2 versus Amplicor HIV-1 DNA test v1.5) for early diagnosis of HIV-1 infection in infants in Senegal

The objective of this study was to compare the performance of the NucliSENS EasyQ HIV-1 v1.2 platform (bioMérieux, France) to the Amplicor HIV-1 DNA test v1.5 (Roche Molecular Systems, Switzerland) in detecting HIV-1 infection in infants using venipuncture-derived whole blood in tubes and dried blood spots. A total of 149 dried blood spots and 43 EDTA-anticoagulated peripheral blood samples were collected throughout Dakar and other areas in Senegal from infants and children aged 3 weeks to 24 months who were born to HIV-1-infected mothers. Samples were tested using the NucliSENS and Amplicor technologies. The NucliSENS and Amplicor results were 100% concordant using either EDTA-anticoagulated peripheral blood or dried blood spots. Compared to Amplicor, the sensitivity and specificity of the NucliSENS test were 100%. The NucliSENS EasyQ HIV-1 RNA assay performed as well as the Amplicor HIV-1 DNA test in detecting HIV-1 infection in infants. In addition, this platform can give an indication of the viral load baseline. The NucliSENS EasyQ platform is a good alternative for early infant diagnosis of HIV-1 infection.     

Expression of decorin and collagens I and III in different layers of human skin in vivo: a laser capture microdissection study

Extracellular matrix (ECM) organization is a complex process that requires the coordinated efforts of many molecules. For the regulation of collagen fiber diameter, the proteoglycan decorin appears to be of major relevance. To investigate the role of decorin in the process of (photo-)aging in more detail, full-thickness punch biopsies were isolated from human buttock skin. Single exposure with two minimal erythemal doses of solar simulated irradiation caused down-regulation of decorin mRNA in young (n = 5) and old subjects (n = 5) after 24 h. Interestingly, decorin mRNA was elevated with age. To test the hypothesis that a decreased collagen-to-decorin-ratio impairs collagen structure we also investigated collagens I and III gene expression. Both were down-regulated with increasing age and after single UV-irradiation. As determined by laser capture microdissection-quantitative real time-Polymerase chain reaction (n = 11), decorin is mostly present in the reticular dermis while being absent from the papillary dermis. Minor expression was also observed in the epidermis. However, in contrast to full-thickness skin biopsies age-dependent changes in collagens I, III, and decorin expression could not be observed with this methodology indicating technical limitations. Together with our finding that collagens I and III mRNA are similarly expressed in the reticular and papillary dermis and are down-regulated by UV, our studies support the idea of a major role of decorin in ECM organization. Altered expression of decorin mRNA in the different dermal strata and a decrease in the collagen-to-decorin ratio inflicted by both age and ultraviolet irradiation possibly affect collagen bundle diameter and subsequently the mechanical properties of human skin.     

Tau protein, phosphorylated tau protein, and beta-amyloid 42 levels in patients with neurodegenerative diseases complicated by cognitive deficits A non-randomized, concurrent, case-control investigation

BACKGROUND： The differential diagnosis of many neurodegenerative disorders depends primarily on clinical symptoms together with imaging methods. Recently, increased importance has been placed on the use of biomarkers for diagnosing various neurodegenerative disorders. OBJECTIVE： To assess the feasibility of tau-protein, phosphorylated tau-protein, beta-amyloid 42 （Aβ42）, and 14-3-3 protein as biomarkers for diagnosing several neurodegenerative diseases complicated by cognitive deficits. DESIGN, TIME AND SETTING： A non-randomized, concurrent, case-control investigation was performed in three medical centers in the Czech Republic （Department of Neurology at the University Hospital in Hradec Kralove, Department of Neurology at the 2rd Medical Faculty, and the University Hospital Motol） between October 2000 and November 2006. PARTICIPANTS： Eighteen patients with probable AIzheimer＇s disease, 4 patients with Creutzfeldt-Jakob disease, 10 patients with frontotemporal dementia, 9 patients with clinically isolated syndrome suggestive of multiple sclerosis, and 7 patients with multiple sclerosis, as well as 38 race-, nationality-, and age-matched cognitively intact controls, were included in the study. Diagnoses were established based on the following criteria： the criteria for Alzheimer＇s disease proposed by the National Institute of Neurological and Communicative Disorders and Stroke/Alzheimer＇s Disease and Related Disorders Association, WHO criteria for Creutzfeldt-Jakob disease, Neary criteria for frontotemporal dementia, and McDonald＇s criteria for multiple sclerosis. All included patients were confirmed to suffer from various degrees of dementia. METHODS： Enzyme-linked immunosorbent assay was used to measure concentrations of tau-protein, phosphorylated tau-protein, and Aβ42 in cerebrospinal fluid （CSF） samples collected by standard lumbar puncture from each patient. Moreover, 14-3-3 protein was assessed by Western blot in CSF of Creutzfeldt-Jakob disease patients. Cognitive status was assessed using the Mini Mental Scale Examination （MMSE） in all subjects. MAIN OUTCOME MEASURES： Establishment of biomarkers with greatest specificity and sensitivity for the investigated disorders according to Receiver Operating Characteristic curves, which were based on values from patients and controls; correlation between concentrations of given biomarkers and demographic parameters, diagnosis, duration of disease, and level of cognitive deficit. RESULTS： Increased concentrations of total tau protein and phosphorylated tau protein, and decreased levels of Aβ42, in CSF of Alzheimer＇s disease patients reached the required sensitivity/specificity ratio of 80% or greater. A marked elevation in CSF concentrations of total tau protein showed even greater sensitivity than 14-3-3 protein in Creutzfeldt-Jakob disease. There was no association between selected biomarkers and frontotemporal dementia or multiple sclerosis. Phosphorylated tau-protein was the only biomarker that noticeably correlated with MMSE scores for Alzheimer＇s disease.CONCLUSION： Levels of total tau protein, phosphorylated tau protein, and A!342 in the CSF could differentiate patients with Alzheimer＇s disease and Creutzfeldt-Jakob disease from healthy controls and patients with other neurodegenerative disorders. The diversity of absolute values demonstrates the necessity to establish a specific standard for each laboratory.     

生物制药方向细胞工程课程建设刍议

分析国内外生物制药方向细胞工程课程的教学情况,根据国内生物制药业发展对细胞工程专业知识的要求和现阶段生物制药人才的实际需求,结合细胞工程研究和应用的内在特点,对生物制药方向的细胞工程课程教学实践提出建议,为生物制药学科建设提供参考。     

双炔失碳酯α、β单体体外抗前列腺癌作用比较

目的比较双炔失碳酯（ANO）的两个差向异构体β、β-ANO体外抗前列腺癌作用的差别。方法氧化铝柱层析法分离纯化仅、B单体,薄层分析法定性检测所分离的产物;在常规培养液中,以4～32μmol·L^-1的β、β-ANO分别作用于人前列腺癌非激素依赖型细胞DU145、PC－3和激素依赖型细胞LIA、LNCaP、RV1,作用时间分别为3d,5d,SRB法测细胞生存率;在去除激素培养液中,以不同浓度的α、β－ANO分别作用于激素依赖型细胞,同时加入雄激素拟似剂（R1881）,SRB法测细胞生存率;以32μmol·L^-1的α、β-ANO分别作用于LNCaP、DU145细胞,相差显微镜观察细胞形态和数量的变化。结果对于非激素依赖型和激素依赖型细胞,α-ANO作用均强于α—ANO,且二者作用差距具有时间和剂量依赖性;α—ANO对抗雄激素的促细胞生长作用强于β—ANO,具有时间和剂量依赖性;相差显微镜观察结果表明,α-ANO作用后,细胞形态及数量变化较之β—ANO更为显著。结论α-ANO体外抗前列腺癌作用强于β—ANO。     

重组人干扰素a-2b栓的质量研究和稳定性研究

目的本次试验研究的目的是将重组人干扰素α-2b制备成通过直肠局部给药达到全身作用的栓剂。方法应用脂肪酸甘油酯suppcioreBM为基质,以Triton X-100为促吸收剂,制备重组人干扰素α-2b栓。以稳定性为指标,考察重组人干扰素α-2b栓的处方。结果用此方法制备的重组人干扰素α-2b栓稳定性强,在2～8℃条件下可保存24个月。结论重组人干扰素α-2b栓的应用可增强患者的顺应性,在临床中应用性强。     

尼莫地平含药脑脊液对体外PC12细胞损伤的保护作用

目的观察尼莫地平含药脑脊液对多种PC12细胞损伤模型的保护作用,对脑脊液药理实验条件的规范化及其应用进行探讨。方法采用噻唑蓝（MTT）法,观察不同取样时间、不同添加量、不同灭活方法处理的尼莫地平大鼠含药脑脊液对氯化钾损伤的PC12细胞存活力的影响,确定尼莫地平大鼠含药脑脊液的制备条件。并进一步考察该条件制备的尼莫地平大鼠含药脑脊液对过氧化氢、连二亚硫酸钠及谷氨酸钠等因素引起的PC12细胞损伤的保护作用。结果尼莫地平含药脑脊液对氯化钾致PC12细胞损伤的保护率随添加量的增加而提高;相同添加量多次给药（2次·d^-1×3.5d）含药脑脊液的保护率均高于单次给药的含药脑脊液;热灭活、乙醇灭活及丙酮灭活含药脑脊液对PC12细胞均有显著保护作用,其保护率与未灭活含药脑脊液无显著性差异;根据确定条件制备的尼莫地平大鼠含药脑脊液对过氧化氢、连二亚硫酸钠及谷氨酸钠等因素损伤的PC12细胞均具有显著的保护作用。结论尼莫地平含药脑脊液的制备以连续多次给药方案,无需灭活处理,体外添加终体积的10%为宜;对氯化钾等多种原因引起的PC12细胞损伤均具有明显保护作用。