益阳橡机通过ISO 14001和OHSAS 18001认证

近日，益阳橡胶塑料机械集团有限公司顺利通过中国质量认证中心ISO14001环境管理体系和OHSAS18001职业健康安全管理体系认证，这是该公司继2000年通过ISO9000质量管理体系认证和2004年橡胶机械产品通过欧洲CE认证后又一次通过的两项认证。     

POY细旦丝上油均匀性的改进

通过油剂调配、输送的工艺优化和设备改造,提高POY上油均匀性。结果表明:通过调配槽的改造,提高了调配油剂乳化均匀性和缩短搅拌时间到30 min左右;通过特殊的高点排气装置,消除气泡引起的POY上油不均和油剂腐败引起的输送不畅。     

硬件设备安全运行与维护策略

服务器运行维护靠的是正确有效的方法和硬件的日常清洁。服务器运行维护技术是通过多年积累而不断成熟的,通过共享而广泛传播的。本文通过介绍服务器运维方法和日常注意事项,让服务器保持高效稳定的状态且发挥它的最大作用。     

碳纤维增强树脂基复合材料性能的研究

以不同含量的二乙烯三胺(DETA)固化的环氧树脂为基体,制备了碳纤维增强树脂基复合材料。通过扫描电镜和红外光谱分析了T-300型碳纤维表面形貌和基团组成。通过拉伸实验、冲击实验对复合材料的力学性能进行了表征;通过紫外老化实验对复合材料的耐候性进行了表征;通过扫描电镜和热重分析对复合材料的断面形貌和耐热性进行了表征。结果表明:碳纤维增强树脂基复合材料具有良好的耐候性、力学性能、而且还具有质量轻、高比强度等一系列优异的性能。     

接枝型含氟丙烯酸酯改性水性聚氨酯的合成

通过溶液聚合的方法合成了端羟基含氟丙烯酸酯大分子,通过HNMR和FT-IR表征了大分子结构,并通过控制链转移剂SHCH2CH2OH用量和加入时间来控制分子量。并以此作为封端剂合成接枝型的含氟丙烯酸酯改性水性聚氨酯(PUA)。通过FT-IR表征了聚氨酯(PUA)的结构。     

矿棉纤维的耐腐蚀性和耐高温性能研究

对炼铁高炉渣通过离心法生产的矿棉纤维进行耐酸碱腐蚀、耐高温性能研究。通过测定酸碱腐蚀前后纤维样品的质量,计算出其质量保留率,同时通过扫描电子显微镜(SEM)对其进行形貌观察和性能研究,探讨其腐蚀机理。通过X射线衍射(XRD)和热重分析(TGA)对高温处理前后的矿棉纤维进行了结构表征和性能测试。结果表明,纤维的耐酸性优于耐碱性,矿棉纤维具有良好的热稳定性能。     

循环水中磷分析方法的探讨

通过条件试验,对测定循环水中无机磷和总磷分析方法进行了改进,优化了操作,分析时间由90min缩短至30min。通过精密度试验和准确度试验,验证了改进后分析方法的准确度。通过方法对比试验,证明改进后的分析方法可用于实际生产。     

滤材多次通过试验装置的优化设计

滤材多次通过试验台关键部位之一是滤材试验装置,它直接决定了试验结果的准确性和可靠性。通过与滤芯多次通过试验结果进行对比,同时结合FLUENT软件对压力及速度分布进行分析,优化了滤材多次通过试验装置的设计。     

基于GSI的安全策略研究

Globus项目通过GSI来提供网格计算环境中的安全认证和安全通信能力。通过对基于GSI的安全技术分析以及任务提交和执行过程的描述，针对各个主体间如何进行身份确认和鉴定的问题，GSI从易用性、灵活性、可移植性和可靠性等方面提出了各种安全策略。通过这些安全策略有效地保证了网格的计算环境的安全性和方便性。     

基于STC单片机的抽油机便携式工况分析仪

以STC12C5A系列单片机为核心设计了一个抽油机便携式工况分析仪,给出了硬件电路设计和软件设计。分析仪通过电压和电流互感器采集抽油机电参数,通过红外光电反射式传感器采集抽油机电机转速,将采集到的数据通过曲线的形式在液晶屏显示并保存到分析仪的存储器上。同时分析仪通过计算给出平衡调整建议。此外,分析仪还可以通过串口与PC实现数据通信,把采集到的数据传输到上位机上。     

An Optimized Protocol for DNA Extraction from Wheat Seeds and Loop-Mediated Isothermal Amplification (LAMP) to Detect Fusarium graminearum Contamination of Wheat Grain

A simple, rapid, and efficient method for isolating genomic DNA from germinated seeds of wheat that is free from polysaccharides and polyphenols is reported. DNA was extracted, treated with RNase, measured and tested for completeness using agarose gel electrophoresis. DNA purification from wheat grains yielded abundant, amplifiable DNA with yields typically between 100 and 200 ng DNA/mg. The effectiveness and reliability of the method was tested by assessing quantity and quality of the isolated DNA using three PCR-based markers. Inter-simple sequence repeats (ISSRs) were used to assess the genetic diversity between different wheat varieties. Specific PCR primer pair Tox5-1/Tox5-2 and a loop-mediated isothermal amplification (LAMP) procedure were used to detect genomic DNA of Fusarium graminearum in contaminated wheat seeds. In this method there is no need to use liquid nitrogen for crushing germinated seedlings. The protocol takes approximately one hour to prepare high quality DNA. In combination with the LAMP assay it is a fast and cost-effective alternative to traditional diagnostic methods for the early detection of toxigenic fusaria in cereals.     

Stepwise Strategy for One-Pot Synthesis of Single-Stranded DNA Rings from Multiple Short Fragments

Cyclic rings of single-stranded (ss) DNA have various unique properties, but wider applications have been hampered by their poor availability. This paper reports a convenient one-pot method in which these rings are efficiently synthesized by using T4 DNA ligase through convergent cyclization of easily available short DNA fragments. The key to the present method is to separate all the splint oligonucleotides into several sets, and add each set sequentially at an appropriate interval to the solutions containing all the short DNA fragments. Compared with simple one-pot strategies involving simultaneous addition of all the splints at the beginning of the reaction, both the selectivity and the yields of target ssDNA rings are greatly improved. This convergent method is especially useful for preparing large-sized rings that are otherwise hard to obtain. By starting from six short DNA fragments (71–82 nt), prepared by a DNA synthesizer, a ssDNA ring of 452-nt size was synthesized in 35 mol % yield and in high selectivity. Satisfactorily pure DNA rings were obtainable simply by treating the crude products with exonuclease.     

壳聚糖与DNA作用的共振光散射特征及微量DNA的光散射测定

通过光谱法研究壳聚糖和核酸的作用 ,结果表明通过共振光散射法用壳聚糖检测微量的 DNA方法可靠、准确且简单。     

应用光敏生物素探针检测Vero细胞乙脑疫苗中细胞残余DNA

用国产光敏生物素,标记Veto细胞全DNA,通过与疫苗中同源DNA的狭缝杂交,检测Vero细胞乙脑疫苗中细胞残余DNA含量。检测灵敏度可达1pg。探针稳定长达一年。杂交反应特:异性高,结果重复性好。与同位素探针相比,生物素标记探针简便、经济、快速、稳定。由本法测得,Vero细胞乙脑粗制疫苗中,含DNA为200～400pg/0.5ml;超滤浓缩苗为4×10~4～6×10~6pg/0.5ml;通过柱层析和沉淀法提纯的疫苗则分别低于60pg/0.5ml和1pg/0.5ml。本法同样适用于传代细胞制备的其它生物制品中细胞残余DNA的检测。     

康柏西普制品中CHO细胞DNA残留量的检测

目的建立特异的CHO细胞DNA残留量荧光定量PCR(Q-PCR)检测方法。方法采用DNeasy Tissue Kit提取CHO细胞基因组DNA,制备DNA标准品;确定Q-PCR反应体系及反应条件,绘制标准曲线,建立CHO细胞DNA残留量Q-PCR检测方法,并验证其专属性、准确性及精密性;用建立的方法对1003b02、1008b05、1012b09、1104b04、1108b13、1112b24批康柏西普原液的CHO细胞DNA残留量进行检测。结果建立的标准曲线Ct值与模板DNA浓度的对数值之间呈良好的线性关系,相关系数为0.99以上;检测HUVEC、HEK293细胞的DNA残留量均无特异性扩增曲线;检测高、中、低浓度的DNA标准品(105、103、101pg/ml)的平均回收率均在Q-PCR方法可接受的回收率范围(50%²00%)内,批间变异系数分别为9.3%、21.8%、26.8%;检测100pg/ml浓度的DNA标准品的平均回收率为111.6%,变异系数为20.4%;康柏西普原液的DNA残留量远低于100 pg/剂量。结论已成功建立特异的CHO细胞DNA残留量Q-PCR检测方法,该方法专属性好,准确性及精密性高,可作为CHO宿主细胞DNA残留量的常规检测方法。     

Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding

Mutagenesis is an important technique for microbial mutation breeding. As the source of mutations, DNA damage extent is a key indicator for the effectiveness of mutagenesis. Therefore, a rapid and easy DNA damage quantification method is required for the comparison of mutagenesis effects and development of mutagenesis tools. Here, we used the umu-microplate test system to quantitatively compare the DNA damage strength caused by atmospheric and room-temperature plasma (ARTP) and other traditional mutagenesis methods including:ultraviolet radiation (UV), diethyl sulfate (DES) and 4-nitroquinoline-1-oxide (4-NQO). The test strain of Salmonella typhimurium TA1535/pSK1002 was used to monitor the time-course profile of β-galactosidase activity induced by DNA damage caused by different mutagenesis methods using a microplate reader. The umu-microplate test results showed that ARTP caused higher extent of DNA damage than UV and chemical mutagens, which agrees well with the result obtained by SOS-FACS-based quantification method as reported previously. This umu-microplate test is accessible for broad researchers who are lack of the expensive FACS instruments and allows the quick quantitative evaluation of DNA damage among living cells for different mutagenesis methods in the study of the microbial mutation breeding.     

CHO宿主细胞DNA残留量检测方法的建立

目的建立检测生物制品中CHO宿主细胞DNA残留量的方法,制备检测CHO细胞DNA残留量的内部参考品,为国家相关标准的制定和修订提供依据。方法分别采用酚:氯仿:异戊醇抽提、DNA提取试剂盒和高盐抽提3种方法提取CHO细胞DNA,比较3种方法的提取效果;用地高辛标记最佳方法提取的CHO细胞DNA探针,并优化标记体系,直接法检测探针的标记效率;采用3种方法处理CHO细胞DNA内部参考品(1组不作处理;2组加入牛血清白蛋白和蛋白酶K;3组按2组方法处理后,增加DNA抽提步骤),通过点杂交法,用标记的探针检测上述样品的灵敏度。结果高盐抽提法提取的CHO细胞DNA效果较好;体系4(DNA10μg,Vial416μl,总体积64μl)标记效率最高;经不同方法处理的1、2、3组CHO细胞DNA内部参考品的检测灵敏度分别为1pg、10pg和1ng。结论建立的地高辛标记CHO细胞DNA探针-点杂交检测CHO细胞DNA残留量的方法稳定可靠,可用于生物制品中CHO宿主细胞DNA残留量的检测。     

重组人肠道病毒71型病毒样颗粒疫苗外源DNA残留量检测方法的建立

目的建立重组人肠道病毒71型(enterovirus 71,EV71)病毒样颗粒(virus-like particles,VLPs)疫苗原液中昆虫细胞宿主DNA残留的地高辛探针杂交检测方法。方法抽提昆虫细胞sf9基因组DNA,经分子筛纯化后作为阳性对照品;分别以1 000~5 000 bp和100~1 000 bp的基因组DNA超声随机小片段为模板,制备地高辛探针,与阳性对照进行杂交试验。同时验证方法的检测范围、灵敏度和特异性,并采用该方法对3批重组EV71 VLPs疫苗原液中宿主DNA残留量进行测定。结果纯化后DNA样品浓度为47. 5 ng/μL,A260/A280值为1. 86,可作为阳性对照。以100~1 000 bp DNA超声随机小片段作为模板,探针标记效率更高,杂交显色更清晰。该法检测范围为10 pg^10 ng,灵敏度达1 pg,特异性强。3批重组EV71 VLPs疫苗原液中每人份剂量的宿主细胞DNA残留均<50 pg。结论本实验建立的方法特异性强,灵敏度高,可用于重组EV71 VLPs疫苗制备过程中的原液检定及质量控制。     

A simple and rapid method for DNA isolation from xylophagous insects

Published methods to isolate DNA from insects are not always effective in xylophagous insects because they have high concentrations of phenolics and other secondary plant compounds in their digestive tracts. A simple, reliable and labor-effective cetyltrimethylammonium bromide-polyvinylpyrrolidone (CTAB-PVP) method for isolation of high quality DNA from xylophagous insects is described. This method was successfully applied to PCR and restriction analysis, indicating removal of common inhibitors. DNA isolated by the CTAB-PVP method could be used in most molecular analyses.     

Surfactant-Assisted Purification of Hydrophobic DNA Nanostructures

Purification of functional DNA nanostructures is an essential step in achieving intended functions because misfolded structures and the remaining free DNA strands in a solution can interact and affect their behavior. However, due to hydrophobicity-mediated aggregation, it is difficult to purify DNA nanostructures modified with hydrophobic molecules by conventional methods. Herein, we report the purification of cholesterol-modified DNA nanostructures by using a novel surfactant-assisted gel extraction. The addition of sodium cholate (SC) to the sample solution before structure folding prevented aggregation; this was confirmed by gel electrophoresis. We also found that adding sodium dodecyl sulfate (SDS) to the sample inhibited structural folding. The cholesterol-modified DNA nanostructures prepared with SC were successfully purified by gel extraction, and their ability to bind to the lipid membrane surfaces was maintained. This method will facilitate the purification of DNA nanostructures modified with hydrophobic molecules and expand their applicability in the construction of artificial cell-like systems.