抗人B7-H1单克隆抗体的制备和鉴定

目的:采用杂交瘤技术制备抗人B7-H1单克隆抗体,并对其进行鉴定。方法:经抗原免疫的小鼠脾细胞与小鼠骨髓瘤细胞以常规方法融合;用间接ELISA法筛选分泌抗体的杂交瘤细胞株;阳性克隆用有限稀释法获得稳定分泌抗人B7-H1单克隆抗体的杂交瘤细胞株;扩增杂交瘤细胞注射进小鼠腹腔后制备腹水;纯化腹水中的单克隆抗体并对其亚型进行鉴定;用间接ELISA法测抗体效价;将肺癌组织制成石蜡切片,用抗人B7-H1抗体进行免疫组化染色。结果:获得1株稳定分泌抗人B7-H1单克隆抗体的杂交瘤细胞株,所分泌的单抗类型为IgG1;抗体效价为1×108,纯化后的抗体含量为6.76g/L;免疫组化实验中,单抗可与肺癌组织表面的B7-H1蛋白特异地结合。结论:制备了人B7-H1单克隆抗体,为B7-H1检测试剂盒的研制奠定了基础。     

关于茭白黑粉菌的正名问题

茭白黑粉菌在文献中出现Yenia esculenta(P.Henn.)Liou及Ustilago esculenta P.Henn.两个名字。现经形态发生学的研究以明确新名字是否有效。此菌冬孢子没有休眠期,孢子成熟后在适宜的环境中即可萌发。萌发的最适pH值为6;最适温度为25％;不需要外界光的诱导,在黑暗条件下也能正常萌发;培养基中的营养成份对孢子萌发的速率有一定影响。冬孢子在不同的环境条件下,其萌发形态及方式都是稳定的:孢子萌发时产生有隔的担子或先菌丝(promycelium)。该先菌丝起初可以是无隔的,但生长到一定程度,便产生分隔,分隔多为2—4个。初生的先菌丝和小孢子是单核的,但成熟的先菌丝、小孢子及短菌丝细胞内均是双核的。可以认为,该菌的双核期在整个生活史中占有相当长的时间,它只有短暂的单核期。没有发现先菌丝、小孢子彼此之间或相互间融合的现象;未见到双核的融合及减数分裂过程。茭白黑粉菌在人工组合培养基上生成厚壁的孢子,较自然界中的冬孢子为大,但萌发方式则相同。最后作者等认为茭白黑粉菌仍应保留在Ustilago属中,而Yenia esculenta(P.Henn.)Liou的名字是无效的。     

Membrane undulation induced by NS4A of Dengue virus: a molecular dynamics simulation study

Nonstructural protein 4A (NS4A) of Dengue virus (DENV) is a membrane protein involved in rearrangements of the endoplasmic reticulum membrane that are required for formation of replication vesicles. NS4A is composed most likely of three membrane domains. The N- and C-terminal domains are supposed to traverse the lipid membrane whereas the central one is thought to reside on the membrane surface, thus forming a u-shaped protein. All three membrane domains are proposed to be helical by secondary structure prediction programs. After performing multi nanosecond molecular dynamics (MD) simulations at various temperatures (300, 310, and 315.15?K) with each of the individual domains, they are used in a docking approach to define putative association motifs of the transmembrane domains (TMDs). Two structures of the u-shaped protein are generated by separating two assembled TMDs linking them with the membrane-attached domain. Lipid undulation is monitored with the structures embedded in a fully hydrated lipid bilayer applying multiple 200?ns MD simulations at 310?K. An intact structure of the protein supports membrane undulation. The strong unwinding of the helices in the domain-linking section of one of the structures lowers its capability to induce membrane curvature. Unwinding of the link region is due to interactions of two tryptophan residues, Trp-96 and 104. These results provide first insights into the membrane-altering properties of DENV NS4A.     

Matrix metalloproteinase-9 is up-regulated by CCL19/CCR7 interaction via PI3K/Akt pathway and is involved in CCL19-driven BMSCs migration

C–C chemokine receptor 7 (CCR7) and its ligands CCL19 contributes to the directional migration of certain cancer cell lines, but its role in the migration of BMSCs remains vague. The aim of this study was to determine the possible interaction between CCL19-induced conditions and matrix metalloproteinases-9 (MMP9) expression in BMSCs. Cell migration using Transwell assay indicated that activation of CCR7 by its specific ligand, exogenous chemokine ligand 19 (CCL19), was associated with a significant linear increase. Western blot and real-time PCR indicated that CCL19/CCR7 significantly upregulated expression of MMP9, which is related to metastasis-associated genes. The CCL19/CCR7 interaction significantly enhanced phosphorylation of Akt, as measured by Western blot. P-Akt and MMP9 protein expression exhibited a time-dependent pattern, and the peak was at 48 h. LY294002 significantly abolished the effects of exogenous CCL19. These results suggest that CCL19/CCR7 contributes to the migration of BMSCs by upregulating MMP9 potentially via the PI3K/Akt pathway.     

固态发酵生产微生物脂肪酶

脂肪酶在水相和非水相中都具有催化活性,在众多工业领域应用前景十分广阔.但脂肪酶的生产成本仍然过高,限制了其在某些工业领域的大规模使用.固体发酵因具有设备比较简单、能耗低、成本低、对环境危害小、易于推广等诸多优点,已逐渐成为微生物脂肪酶生产的一个重要方式.由于能源成本的抬高和人们环保意识的加强,自上世纪90年代以来,原来一直认为技术含量较低的固态发酵技术重新受到重视并得到了快速的发展.综述了固态发酵在脂肪酶生产中的应用研究,重点介绍了固态发酵生产脂肪酶的特点、脂肪酶固态发酵的影响因素及其生物反应器.     

Characterization and potential function of a novel pre‐implantation embryo‐specific RING finger protein: TRIML1

Members of the super‐class of zinc finger proteins are key regulators in early embryogenesis. Utilizing in silico mining of EST Databases for pre‐implantation Embryo‐Specific Zinc Finger Protein Genes, we characterized a novel zygotic mouse gene—tripartite motif family‐like 1 (TRIML1), which expresses in embryo before implantation. Knocking down of TRIML1 resulted in the fewer cell number of blastocysts and failture to give rise to neonates after embryo transfer. The binding partner of TRIML1, Ubiquitin‐specific protease 5 (USP5), was identified by yeast two‐hybrid screening assay. The interaction was confirmed by GST pull‐down and coimmunoprecipitation analysis. The role of TRIML1 in ubiquitin pathway during the development stage of mouse blastocyst was further discussed. Mol. Reprod. Dev. 76: 656–664, 2009. © 2009 Wiley‐Liss, Inc.     

光诱导角膜交联及检测技术的研究进展

由于圆锥角膜疾病导致越来越多的人患有近视,常见的矫正方法有佩戴近视眼镜、隐形眼镜等.随着科技的进步,利用光对近视等眼科疾病进行屈光矫正已经成为当前临床中常用的方法.使用光诱导角膜胶原蛋白发生交联,从而达到治疗圆锥角膜疾病、提高患者视力水平的目的,这是一种新型的光治疗眼睛疾病的方法.同时这种方法由于无侵入性、对操作者能力依赖性小等优势成为新的研究热点.本文阐述光诱导角膜交联的基本原理,并介绍其发展历程,分析现有的各种交联方法和角膜检测技术的原理,并对现有交联方法和检测方法的优缺点进行讨论.最后,本文对光诱导角膜交联和检测技术的最新进展进行系统的论述,并对未来的发展趋势进行展望.     

磷模型在千岛湖水体污染预测中的应用研究

运用国际上通用的多个经验统计磷模型预测千岛湖目前的磷浓度水平,其中第6个模型最适合用于千岛湖的预测预报．利用该模型预测了磷负荷量分别减少1/4、1/2、3/4情况下,千岛湖水体中总磷浓度的响应．同时针对千岛湖营养水平控制的短、长期目标,利用模型计算出了所允许的总磷负荷量及入湖磷浓度．     

合肥秋冬季茶园天敌对假眼小绿叶蝉和茶蚜的空间跟随关系

为了合理保护和利用天敌及科学地选取抽样方法,开展了合肥地区秋冬季4个品种茶园假眼小绿叶蝉Empoasca vitisGothe和茶蚜Toxoptera aurantii Boyer与其天敌之间空间关系研究,运用地学统计学方法求得天敌和害虫各自的变程,用灰色关联度分析方法分析害虫与天敌变程的关联度,关联度值越大的天敌在空间上对害虫的跟随关系越密切。分析了2010年9月28日至11月25日期间假眼小绿叶蝉和茶蚜数量最少的舒茶早茶园和二种害虫数量最多的平阳特早茶园天敌对害虫空间上的跟随关系,结果表明,二种茶园的假眼小绿叶蝉和茶蚜及其4种主要天敌均为聚集分布,舒茶早茶园与假眼小绿叶蝉空间上跟随关系密切的前二位天敌是斜纹猫蛛Oxyopes sertatus L.Koch(0.8594)和草间小黑蛛Erigonidium graminicolum Sundevall(0.8397),与茶蚜空间上跟随关系密切的前二位天敌是草间小黑蛛(0.7448)和斜纹猫蛛(0.7433);平阳特早茶园与假眼小绿叶蝉空间上跟随关系密切的前二位天敌是八斑球腹蛛Theridion ocomaculatum Bose.et Str(0.8207)和斜纹猫蛛(0.8104),与茶蚜空间上跟随关系密切的前二位天敌是八斑球腹蛛(0.8324)和斜纹猫蛛(0.7730)。其中,11月25日4种茶园假眼小绿叶蝉和茶蚜数量均较多。分析了该日另外二个茶树品种福云六号和龙井长叶茶园假眼小绿叶蝉和茶蚜与其天敌的空间关系,结果表明,二种茶园假眼小绿叶蝉和茶蚜及其天敌均为聚集分布,福云六号茶园与假眼小绿叶蝉变程值(3.8182)最接近的天敌是斜纹猫蛛(4.7222),与茶蚜变程值(6.5854)最接近的天敌是斜纹猫蛛(4.7222);龙井长叶茶园与假眼小绿叶蝉变程值(1.0000)最接近的天敌是八斑球腹蛛(1.0000),与茶蚜变程值(4.5000)最接近的天敌是斜纹猫蛛(7.6316)。总之,秋冬季4个品种茶园斜纹猫蛛在空间上是假眼小绿叶蝉和茶蚜跟随关系最密切的天敌,其次是八斑球腹蛛和草间小黑蛛。