SPA方法快速检验食品志贺氏菌的研究

ＳＰＡ是某些金黄色葡萄球菌细胞壁的一种蛋白质成分，具有同人和多种哺乳动物ＩｇＧ分子Ｆｃ段结合的能力。本方法用含Ａ蛋白丰富的金黄色葡萄球菌菌体与志贺氏菌属抗血清在一定条件下混合致敏，让抗体结合到金黄色葡萄球菌细胞壁Ａ蛋白上，制成ＳＰＡ诊断试剂。     

人热休克蛋白73的基因重组、表达和纯化

目的　用基因重组的方法表达人热休克蛋白７３,并纯化表达产物,用于进一步分析。方法　用人 热休克类似蛋白ＨＳＰ７３基因构建原核细胞表达载体ｐＥＴＨＳＰ７３,在大肠杆菌ＢＬ２１中用半乳糖苷（ＩＰＴＧ）诱导表达,用 电泳法和 ＡＴＰ亲和层析法提取 ＨＳＰ７３。经ＳＤＳ－ＰＡＧＥ、用３ ａ３抗体 Ｗｅｓｔｅｒｎ ｂｌｏｔ作 ＥＣＬ检测。结果人 ＨＳＰ７３基因 构建的载体ｐＥＴＨＳＰ７３能很好地在大肠杆菌表达出相对分子质量为７３　０００的蛋白。两种蛋白提取方法均能有效提纯表达的蛋白,该蛋白具有人ＨＳＰ７３抗原特性。所得蛋白质氨基酸测定和Ｎ端测序的结果与有关的报道一致。结 论　ＨＳＰ７３基因重组、表达和纯化方法的建立,为研究ＨＳＰ７３的结构、功能与临床应用提供了必要条件。     

A群脑膜炎球菌多糖结合疫苗的研制

目的 制备安全有效的Ａ群脑膜炎球菌多糖结合疫苗。方法 通过碳二亚胺（ＥＤＡＣ）将Ａ群脑膜 炎球菌多糖－ＡＨ衍生物与破伤风类毒素（ＴＴ）蛋白共价结合,制备Ａ群脑膜炎球菌多糖结合疫苗。结果 所结合 成的多糖－蛋白结合物,具有Ａ群脑膜炎球菌多糖和破伤风类毒素抗原特异性。单独用多糖免疫小鼠未能诱导出 高水平的血清Ｐｓ抗体,而用结合物免疫小鼠却诱导出高水平的血清Ｐｓ抗体及ＴＴ抗体,并有免疫记忆性。结论 为进一步临床评价该疫苗的人群免疫效果提供了实验基础。     

聚乙烯醇纤维的形态结构及性能研究

用ＳＥＭ,ＩＲ,ＤＳＣ等研究了聚醋酸乙烯（ＰＶＡｃ）醇解纺丝得到的聚乙烯醇（ＰＶＡ）纤维的形态结构、分子结构及热性能。用纤维电子强伸仪研究了纤维的力学性能。证明由ＰＶＡｃ醇解纺丝可得到强度和模量分别为１３．２７ｃＮ／ｄｔｅｘ和３５０．１０ｃＮ／ｄｔｅｘ的高性能ＰＶＡ纤维。     

反应型增容剂PS-co-GMA的合成及反应动力学的研究

以过氧化苯甲酰（ＢＰＯ）为引发剂,利用改进的本体聚合方法制备了苯乙烯—甲基丙烯酸环氧丙酯共聚物（ＰＳｃｏＧＭＡ）,用红外光谱证实了ＰＳｃｏＧＭＡ共聚物的存在,并探讨了聚合温度,聚合时间对聚合物的分子量和转化率的影响,通过以上研究确定了最佳反应条件：反应温度为７０℃,反应时间为６ｈ时,可得到性能满意的ＰＳｃｏＧＭＡ共聚物。     

乙酸乙烯酯嵌段共聚物的合成研究

以ＡＩＢＮ为引发剂，用ＣＣｌ４调节乙酸乙烯酯聚合制备了带三氯甲基端基的ＰＶＡｃ大分子引发剂（ＰＶＡｃ－ＣＣｌ３）。以ＣｕＣｌ－ｂｐｙ为催化剂，用ＰＶＡｃ－ＣＣｌ３引发Ｓｔ，ＭＭＡ和ＢＭＡ等单体的ＡＴＲＰ聚合，得到了一系列相对分子质量可控、相对分子质量分布较窄的ＰＶＡｃ嵌段共聚物。     

重组人巨细胞病毒gp52蛋白的纯化及鉴定

目的 对大肠杆菌表达的人巨细胞病毒ｇｐ５２蛋白纯化及鉴定。方法 将表达ｇｐ５２蛋白的菌体超 声裂解后,离心取上清,经ＤＥＡＬＥ－Ｓｅｐｈａｒｏｓｅ ＦＦ阴离子柱纯化,收集纯化的ｇｐ５２蛋白,再上Ｓ－Ｓｅｐｈａｒｏｓｅ　ＦＦ阳离子柱纯 化,收集纯化的ｇｐ５２蛋白,经ＳＤＳ－ＰＡＧＥ检测其纯度,用Ｗｅｓｔｅｍ　ｂｌｏｔ和 ＥＩＬＳＡ检测其抗原性。结果 纯化的ｇｐ５２蛋 白纯度达８５％以上,并有较好的抗原性和特异性。结论 制备的重组ｇｐ５２蛋白可用于人巨细胞病毒抗体的检测 等研究。     

百日咳疫苗血清学效力试验与小鼠保护力试验结果的相关性

目的 了解百日咳疫苗血清学效力试验（ＰＳＰＴ）与小鼠保护力试验（ＭＰＴ）结果的相关性,为该法替代ＭＰＴ积累资料。方法 用不同剂量的ＤＴＰ疫苗免疫小鼠,４ｗ后采血,用ＥＬＩＳＡ测定小鼠血清中抗百日咳菌表面 抗原的总抗体滴度。用平行线法计算疫苗的效力,与ＭＰＴ测定结果进行相关性分析。结果１５批疫苗分别用 ＰＳＰＴ和ＭＰＴ法测定的效力单位相关性显著（ｒ＝ ０．９３５８,ｐ＜０．００１）,二者差异无显著意义（Ｐ＞０．０５）。与 ＭＰＴ法相 比,ＰＳＦＦ法有更高的重复性,而且９５％可信限较小。结论ＰＳＰＴ法有望取代ＭＰＴ。     

γ-ray引发的VAc种子乳液聚合的研究

用辐射法制备了以ＰＳｔ为核以ＰＶＡｃ为壳的乳胶粒子。电观察结果表明,种子聚合后乳胶粒子半径增加８倍。对反应的机理及动力学进行了研究,为ＰＶＡｃ乳液的改性作了基础实验方面的探索研究 。     

一项微生物能力验证的多种方法检测结果分析

为确保微生物能力验证结果的准确报出,本实验室采用了多种方法进行CNAS T 0710的铜绿假单胞菌(Pseudomonas aeruginosa)和金黄色葡萄球菌(Staphylococcus aureus)的能力验证比对检验,结果表明:以上多种检测方法的测试结果相吻合,均显示5份样品中有2份检出铜绿假单胞菌,2份检出金黄色葡萄球菌。最终5份样品测试均取得满意结果,在分析比较中也总结出各种检验方法的优缺点。     

Zinc(II) coordination complexes as membrane-active fluorescent probes and antibiotics

Molecular probes with zinc(II)-(2,2'-dipicolylamine) coordination complexes associate with oxyanions in aqueous solution and target biomembranes that contain anionic phospholipids. This study examines a new series of coordination complexes with 2,6-bis(zinc(II)-dipicolylamine)phenoxide as the molecular recognition unit. Two lipophilic analogues are observed to partition into the membranes of zwitterionic and anionic vesicles and induce the transport of phospholipids and hydrophilic anions (carboxyfluorescein). These lipophilic zinc complexes are moderately toxic to mammalian cells. A more hydrophilic analogue does not exhibit mammalian cell toxicity (LD(50) >50 microg mL(-1)), but it is highly active against the Gram-positive bacteria Staphylococcus aureus (MIC of 1 microg mL(-1)). Furthermore, it is active against clinically important S. aureus strains that are resistant to various antibiotics, including vancomycin and oxacillin. The antibiotic action is attributed to its ability to depolarize the bacterial cell membrane. The intense bacterial staining that was exhibited by a fluorescent conjugate suggests that this family of zinc coordination complexes can be used as molecular probes for the detection and imaging of bacteria.     

Functionalisation of Inorganic Material Surfaces with Staphylococcus Protein A: A Molecular Dynamics Study

Staphylococcus protein A (SpA) is found in the cell wall of Staphylococcus aureus bacteria. Its ability to bind to the constant Fc regions of antibodies means it is useful for antibody extraction, and further integration with inorganic materials can lead to the development of diagnostics and therapeutics. We have investigated the adsorption of SpA on inorganic surface models such as experimentally relevant negatively charged silica, as well as positively charged and neutral surfaces, by use of fully atomistic molecular dynamics simulations. We have found that SpA, which is itself negatively charged at pH7, is able to adsorb on all our surface models. However, adsorption on charged surfaces is more specific in terms of protein orientation compared to a neutral Au (111) surface, while the protein structure is generally well maintained in all cases. The results indicate that SpA adsorption is optimal on the siloxide-rich silica surface, which is negative at pH7 since this keeps the Fc binding regions free to interact with other species in solution. Due to the dominant role of electrostatics, the results are transferable to other inorganic materials and pave the way for new diagnostic and therapeutic designs where SpA might be used to conjugate antibodies to nanoparticles.     

葡萄球菌蛋白A的纯化新工艺

目的建立一种高效、简便、实用的分离纯化葡萄球菌蛋白A(SPA)的工艺。方法采用超声波破菌和IgG-Sepharose4B亲和层析分离纯化SPA;以Lowry法检测蛋白含量;双向琼脂免疫扩散法测定效价和特异性;用还原和非还原SDS-PAGE法检测纯度和相对分子质量。结果此法制得的3批SPA的蛋白含量分别为4·7、4·4和5·4mg/ml,收率分别为2·70、2·64和3·78g/100g湿菌。对人IgG的免疫双扩散效价均为1:64;与正常人、豚鼠、家兔和小鼠的血清反应均出现一条沉淀线,而与鸡和羊不出现沉淀线。经非还原SDS-PAGE检测只呈现一条蛋白带,相对分子质量约为160000～180000;经还原SDS-PAGE检测呈现两条蛋白带,相对分子质量分别约为67000和34000。结论已成功建立了分离纯化SPA的新工艺。     

Mikrobiologische Untersuchungen an ozonisiertem Olivenöl

Microbiological Investigations with Ozonized Olive Oil The general bactericide effect of ozonized olive oil was proved for various strains of bacteria. The most significant destruction effect was found for staphylococcus aureus; cultures of this strain, 24 hours old, showed a relative destruction effect of 99.9% after application of 1% inhibitor concentration with 90 min. duration of action. It was found that the staphylococcus aureus cells during the exponential growth of the period have a decadic rate of dying-back of about one per hour. The destruction effects, caused by freshly ozonized olive oil, differed only to a low extent from those of the ozonized olive oil stored for eight years.     

抗人IgG-抗HRP双特异性单克隆抗体的研制及初步应用

用8－AG（8－氮杂鸟嘌呤）驯化的HRP－MCAb（抗辣根过氧化物酶单克隆抗体）杂交瘤细胞HAT（次黄嘌呤氨基喋呤胸苷）敏感株与人IgG免疫的BALB／c小鼠脾细胞进行二次融合，获得5株能稳定分泌抗人IgG－抗HRP的BsMcAb（双特异性单克隆抗体）杂交－杂交癌细胞，用其混合腹水经HRP亲和层析纯化的BsMcAb制备BsMcAbPAP（双特异性单抗过氧化物酶-抗过氧化物酶复合物）试剂代替丙肝诊断试剂盒中酶标二抗，比较结果初步表明两者差异无显著意义，将为IgG的检测提供有用试剂。     

淋病SPA协同凝集快速诊断试剂的研究

采用吸附解离法获得高度纯化的抗淋球菌血清,用以致敏金葡菌Cowan—Ⅰ株,制成诊断淋病用试剂。本试剂与多种其他细菌及多株淋球菌作协同凝集试验,证明具有高度的特异性和敏感性。可直接用于检测患者尿(阴)道分泌物中的淋球菌或其抗原成分,经110例急慢性淋病患者检材试用,阳性率达94.55％,显著优于培养法(61.82％)。     

A Universal Fluorescent Immunochromatography Assay Based on Quantum Dot Nanoparticles for the Rapid Detection of Specific Antibodies against SARS-CoV-2 Nucleocapsid Protein

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the pathogenic agent leading to COVID-19. Due to high speed of transmission and mutation rates, universal diagnosis and appropriate prevention are still urgently needed. The nucleocapsid protein of SARS-CoV-2 is considered more conserved than spike proteins and is abundant during the virus’ life cycle, making it suitable for diagnostic applications. Here, we designed and developed a fluorescent immunochromatography assay (FICA) for the rapid detection of SARS-CoV-2-specific antibodies using ZnCdSe/ZnS QDs-conjugated nucleocapsid (N) proteins as probes. The nucleocapsid protein was expressed in E.coli and purified via Ni-NTA affinity chromatography with considerable concentration (0.762 mg/mL) and a purity of more than 90%, which could bind to specific antibodies and the complex could be captured by Staphylococcal protein A (SPA) with fluorescence displayed. After the optimization of coupling and detecting conditions, the limit of detection was determined to be 1:1.024 × 10⁵ with an IgG concentration of 48.84 ng/mL with good specificity shown to antibodies against other zoonotic coronaviruses and respiratory infection-related viruses (n = 5). The universal fluorescent immunochromatography assay simplified operation processes in one step, which could be used for the point of care detection of SARS-CoV-2-specific antibodies. Moreover, it was also considered as an efficient tool for the serological screening of potential susceptible animals and for monitoring the expansion of virus host ranges.     

真养产碱杆菌的化学破胞过程

１引言近年来，利用真养产碱杆菌（Ａｌｃａｌｉｇｅｎｅｓｅｕｔｒｏｐｈｕｓ）生产生物可降解高分子聚？羟基丁酸酯（ｐｏｌｙβｈｙｄｒｏｘｙｂｕｔｙｒａｔｅ，ＰＨＢ）的研究已引起人们的极大兴趣，其中出现了很多破碎真养产碱杆菌的技术［１］。前人研究了机械...     

Efficiency enhancement of graphene/silicon-pillar-array solar cells by HNO3 and PEDOT-PSS

A single-layer graphene film was grown on copper foil by chemical vapor deposition and transferred onto a silicon-pillar-array (SPA) substrate to make a Schottky junction solar cell. The SPA substrate was specifically designed to suppress reflectance and enhance light absorption. The energy conversion efficiency of the prepared graphene/SPA solar cells achieved a maximum of 2.90% with a junction area of 0.09 cm(2). HNO(3) was employed to dope the graphene in the solar cells, and the time dependence of HNO(3) treatment on the cell performance was studied. Poly(3,4-ethylenedioxythiophene) polystyrenesulfonic acid (PEDOT-PSS) was also introduced in graphene/SPA solar cells by spin coating on top of the graphene film, and its modification on the cell performance was characterized. The results show that both HNO(3) and the PEDOT-PSS film could enhance the energy conversion efficiency of graphene/SPA solar cells.     

化妆品防腐剂的短期抑菌效能评价

分别将金黄色葡萄球菌、大肠杆菌的菌悬液与适当浓度的杰马BP、GVL和尼泊金甲酯（以下简称尼甲）3种防腐剂的水溶液或添加防腐剂的化妆品混合均匀,作用4h、8h和24h后进行菌落总数测定,以细菌总数的杀灭对数值作为评价指标。结果显示,尼甲按1：1000-3：1000的比例加入水中后,对大肠杆菌、金黄色葡萄球菌的抑菌作用弱。浓度提高到5：1000～15：1000加入化妆品中后,对大肠杆菌的抑菌效果明硅增强．但对金黄色葡萄球菌的抑杀作用反而减弱。在膏状化妆品中尼甲对大肠杆菌的抑杀作用明显强于液状化妆品。GPL和杰马BP在0．05％～0．15％的水溶液中对2种致病菌产牛明显甚至强烈的抑杀作用,且对2种菌的抑杀强度基本相同,浓度提高到O．25％～0．75％并加入化妆品中后,抑菌效果明显下降。在膏状化妆品中GPL和杰马BP对大肠杆菌的抑杀作用强于金黄色葡萄球菌。膏状和液状化妆品中GPL和杰马BP对大肠杆菌的抑杀作用基本相当。