基因工程菌E.Coli2426／pMN表达产物的纯化

用基因工程菌Ｅ．Ｃｏｌｉ２４２６／ｐＭＮ高效表达了麦芽糖结合蛋白—人神经生长因子（β）融合蛋白，菌体经超声波破碎后，上清液经Ａｍｙｌｏｓｅ亲和柱一步即可获得ＳＤＳ－ＰＡＧＥ纯蛋白质，回收率为１０％左右，按标准分子量计，该融合蛋白（５５ＫＤａ）确是麦芽糖结合蛋白（４２ＫＤａ）与人神经生长因子（β）（１３ＫＤ）的络合物     

神经生长因子

概述了神经生长因子的分子结构及作用机制，综述了神经生长因子的生理功能，神经生长因子不仅对神经细胞的存活，生长发育，分化，再生和功能维持起调控作用。同时也能诱导细胞凋亡。     

神经生长因子有助修复受损心肌

[科技日报]英国布里斯托尔人学的一项研究发现，提升心脏病患者心脏部位的神经生长因子（NGF）水水平，可帮助受损心肌快速修复，从而有效提高患者的生存几率。这一研究结果发表在最近出版的《循环研究》杂志上。     

神经的生长与再生

神经的生长与再生与胚胎学极其相似.在胚胎期神经的诱导是与可弥散的物质有关,这些物质可称为神经生长因子.神经的存活和生长是离不开神经生长因子的.在神经轴突膜上有一个能识别并能结合神经生长因子分子的特异受体,从而导致神经生长因子分子促使轴突生长和维持神经元存活起生化活动的作用.成年个体的神经损伤以后,在受伤的部位总要产生一些胶状渗出物,这些物质对损伤的神经起诱导作用,从而诱导神经再生.这就是神经的诱导性.     

分光光度法测定盐源山蛭素对凝血酶的抑制活性

报告了一实用的凝血酶抑制剂抑制活性的测定方法——发色底物分光光度法；以发色底物S—2238与标准凝血酶反应，测定未抑制的吸光度Ao，同时以发色底物S—2238与凝血酶抑制剂盐源山蛭素和标准凝血酶反应测定抑制吸光度As，然后以As／Ao计算盐源山蛭素的抑制活性a1．实验证明，这种对盐源山蛭素抑制活性的测定方法简便易行、重复性高、偶然误差小、可信度高．     

酚类化合物苯环碳原子的化学位移与其净电荷的关？…

用ＡＢ－ＩＮＩＴＩＯ法对只含酚羟基的７种酚化合物的苯环碳原子的净电荷做了计算，并测定或查阅了它们的＾１３ＣＮＭＲ谱，参考标准图谱找出了Ｃｉ对应的化学位移δｉ，发现Ｑｉ－δｉ间呈现较好的线性关系。每种化合物均有最负净电荷与最低化学位移（δｍｉｎ），Ｑｍｉｎ与δｍｉｎ出现在酚羟在邻位同一碳原子上，而这一碳原子具有最大的亲核－亲电反应活性。那些δｉ〉１３０的碳原子反应活性极小，以至不能发生亲核－亲电反应     

茯苓化学成分及生理活性研究

1:利用PC12细胞测定生理活性,以萃取物的生理活性为导向,采用硅胶柱色谱和高效液相色谱法分离纯化茯苓的Me OH提取物中的活性成分,并通过光谱数据解析对化合物进行结构鉴定。分离得到了7个羊毛甾烷型三萜化合物,分别为多孔蕈酸C(1),去氢松苓酸(2),茯苓酸(3)氢化松苓酸(4)去氢松苓酸甲酯(5)去氢茯苓酸(6)和土莫酸(7)。化合物1,2,5~7在0.3~30.0μM浓度范围内,对神经生长因子诱导的PC12细胞显示出神经伸展促进活性。同时,构效关系讨论结果表明7位和9位上连接的二烯键是这类化合物显示活性的关键基团。     

电炉钢渣活性激发研究

对电炉钢渣（熔炼渣和精炼渣）在机械激发、热激发和化学激发作用下的活性特征进行研究．研究表明：标准养护条件下，电炉钢渣活性指数随比表面积增加而显著增加；热激发作用下，电炉钢渣活性指数有较大幅度提高；热激发作用下钢渣的活性指数也随着比表面积的增加而提高；常温养护和蒸压养护条件下，精炼渣都没有表现出比熔炼渣更高的活性；无论是标准养护还是蒸压养护，电炉钢渣在各种碱存在的情况下，大多表现为强度下降，仅Na2s0。或NaOH在蒸压养护条件下对熔炼渣有激发作用，提高了熔炼渣的活性．     

老年痴呆症的药物治疗进展

老年痴呆症（ＡＤ）是一种神经退行性疾病，至今病因未明，且尚无特效药，近年来，临床运用促智药、胆碱酯酶抑制剂、乙酰胆碱释放增强剂、ＮＳＡＩＤ、神经生长因子等药物，对改善症状、延缓疾病表一定的作用。此病的病因及药的治疗有待作更深入的研究 。     

低分子量肝素的制备及其结构与活性差异

低分子肝素作为一种抗血栓的多糖药物在临床中已应用了二十多年，目前已作为外科预防血栓形成药物，并在治疗急性静脉栓塞紊乱方面取代了末分级肝素。因肝素的来源和制备的方法不同使低分子肝素的精细结构不同，低分子肝素结构的复杂性，使得各产品的生物活性，例如抗蛋白酶活性不同，从而导致其临床使用的标准不同。该文将对低分子肝素的制备方法及其结构和抗蛋白酶活性的差异进行报导。     

NGF诱导PC12细胞神经元样细胞分化

目的:建立PC12细胞的神经元样细胞分化模型,并探讨ERK蛋白在PC12细胞神经元样细胞分化中的可能机制.方法:以10、20、50 g/L的NGF(NGF溶于PBS,培养基中PBS的终浓度不超过2%)培养PC12细胞,应用倒置相差显微镜、显微镜测微尺及流式细胞仪鉴定PC12细胞的分化,以确定NGF使用剂量.运用免疫印迹检测不同浓度、不同作用时间时ERK蛋白在PC12细胞中的表达,并进行统计学分析.结果:随NGF剂量的升高,PC12细胞的体积、最长突起长度和突起数目均会增大或增多。统计学分析证明50 g/L的NFG作用48 h足以诱导PC12细胞的交感神经样改变。尽管ERK总蛋白水平在NFG作用前后无明显改变,但在NFG作用5 min后磷酸化的ERK蛋白水平即显著升高,达到峰值,并持续约1 h.50 g/L的NFG作用于PC12细胞48 h可使细胞发生明显的G1期阻滞.结论:PC12细胞可在NGF作用下出现交感神经样改变,并且细胞的分化程度依赖于NGF的使用剂量和作用时间.在NGF诱导的PC12细胞神经元样分化中ERK蛋白的磷酸化对NGF呈现剂量和时间依赖性,提示ERK蛋白在分化的早期发挥重要作用.     

多壁碳纳米管-神经生长因子复合物的制备及生物活性测定

为探讨采用羧基化多壁碳纳米管（MWCNTs—COOH）非共价接枝神经生长因子（NGF）制备碳纳米管神经生长因子（MWCNTsNGF）复合物,考察复合物的生物活性。采用透射电子显微镜（TEM）表征MWCNTs—NGF复合物的微观形貌,酶联免疫吸附法（ELISA法）测定MWCNTs—NGF复合物载带NGF的量,MTT法测定了MWCNTs—NGF复合物的对嗜铬细胞瘤细胞（PCI2细胞）的毒性,PCI2细胞培养法评价复合物的生物活性,TEM表征复合物与细胞的分布情况。结果：TEM图像表明NGF连接到了MWCNT上,EI．ISA法测得MWCNTsNGF复合物载带NGF的量为797．63pg／mg,MWCNTs—NGF复合物对PCI2细胞有一定的毒性,生物活性试验表明NGF浓度相同的情况下,MwCNTs—NGF复合物组PCI2细胞的分化率明显高于NGF组。TEM图像表明碳纳米管能进入细胞。结论：碳纳米管能载带NGF进入细胞,使NGF能更好的表达生物活性。     

Sarcoma viruses carrying ras oncogenes induce differentiation-associated properties in a neuronal cell line

The growth-promoting and/or differentiation-blocking activities of Kirsten (Ki-MSV) or Harvey murine sarcoma virus (Ha-MSV) on various types of cells in vitro are well documented. Here we report an unexpected effect of these viruses on a rat phaeochromocytoma cell line, PC12. PC12 cells, which multiply indefinitely in growth medium, are known to respond to nerve growth factor (NGF) by cessation of cell division and expression of several properties resembling those of differentiated sympathetic neurones. We have found that Ki- and Ha-MSV mimic some, if not all, of the activities of NGF in PC12 cells, and there is evidence that the viral oncogenes, v-Ki-ras and v-Ha-ras, are responsible for this phenomenon. This system may be of value for studying the mechanism of action of the v-ras genes as well as the regulatory mechanism of growth and differentiation in neuronal cells.     

Role of ion flux in the control of c-fos expression

There has been much interest in the biochemical and biophysical processes that couple extracellular signals to alterations in gene expression. While many early events associated with the treatment of cells with growth factors have been described (for example, ion flux and protein phosphorylation), it has proved difficult to establish biochemical links to gene expression. Recently, the study of such genomic control signals has been facilitated by the demonstration that the c-fos proto-oncogene is rapidly and transiently induced by treatment of several cell types with polypeptide growth factors and other growth modulating substances. In one particular system it has been shown that nerve growth factor (NGF) causes a transient induction of c-fos in the phaeochromocytoma cell line PC12, within 15 min. Furthermore, the magnitude of this induction can be modulated with pharmacological agents such as peripheral-type benzodiazepines (BZDs). Thus, the study of c-fos expression in PC12 cells could yield valuable clues to the coupling mechanisms linking cell surface activation to genomic events. Here we demonstrate that c-fos is induced in PC12 cells either by receptor-ligand interaction or by agents or conditions that effect voltage-dependent calcium channels.     

红藻氨酸致癫痫大鼠不同时点神经生长因子变化研究

摘要: 目的 探讨红藻氨酸( Kainic Acid,KA) 致癫痫大鼠癫痫发作过程中海马组织神经生长因子( nerve growth factor,NGF) 不同时间点表达量的变化。方法 腹腔注射 KA 建立大鼠癫痫模型。采用 TaqMan 探针实时定量 PCＲ ( Ｒeal-time quantitative PCＲ) 技术,检测注射 KA 后不同时点大鼠海马组织中 NGF 表达量的变化。结果 与 NGF 表达量与生理盐水对照组( NS) 相比,6 ～ 12 h NGF 表达量明显低于 NS 组( P ＜ 0. 05) 、48 ～ 72 h NGF 表达量开始升高并高于 NS 组,在 24、72 h 时其表达量显著高于 NS 组( P ＜ 0. 05) 。结论 NGF 对致癫大鼠的海马具有修复与保护作用。     

脑内移植NGF诱导的PC-12细胞治疗大鼠帕金森病的研究

实验利用SD大鼠复制6-羟多巴胺(6-hydroxydopamine,6-OHDA)完全损伤型帕金森动物模型,将神经生长因子(nerve growth factor,NGF)处理后的肾上腺嗜铬细胞瘤细胞(plleoclxromocytoma cells,PC-12)移植人模型大鼠纹状体内,观察模型动物行为改善和移植PC-12细胞的存活情况.经行为学检测,6-OHDA动物模型复制成功率达55.1%.免疫组织化学、蛋白印迹检测结果显示模型动物黑质多巴胺能神经元数目减少,黑质酪氨酸羟化酶含量降低证明模型动物稳定、可靠.PC-12细胞经NGF(50 μg/L)连续诱导7 d,细胞逐渐平展、细胞膜变皱褶等神经元形态学特征出现后,于纹状体内行细胞移植术,经阿普吗啡(apomorphine,APO)诱导旋转行为有明显改善,蛋白印迹检测也发现移植侧具有显著的酪氨酸羟化酶免疫阳性信号.因此,NGF诱导后的PC-12细胞可以作为治疗帕金森的一种细胞供体.     

Inhibition of growth factor-induced differentiation of PC12 cells by microinjection of antibody to ras p21

The protein products (p21) of the ras cellular proto-oncogenes are thought to transduce membrane signals necessary for the induction of cell division. However, there is uncertainty as to the precise role of ras p21 in mediating ligand-membrane receptor signals leading to cell differentiation. Treatment of rat phaeochromocytoma cells (PC12) with nerve growth factor (NGF) results in the induction of a number of phenotypic characteristics of sympathetic neurones, including cessation of cell division and outgrowth of neuronal processes (neurites). Here we report that microinjection of antibody to ras p21 into PC12 cells inhibited neurite formation and resulted in temporary regression of partially extended neurites, an effect which was observed up to 36 h after initiation of NGF treatment. Neurite formation induced by cyclic AMP was unaffected by injection of anti-p21 antibody. These results indicate that p21 is involved in the initiation phase of NGF-induced neurite formation in PC12 cells and has a role in hormone-mediated cellular responses distinct from cell proliferation.     

Differentiation of PC12 phaeochromocytoma cells induced by v-src oncogene

PC12 rat phaeochromocytoma cells are a model system that can be used to study both neuronal differentiation and the mechanism of action of nerve growth factor (NGF). PC12 cells respond to NGF protein by shifting from a chromaffin-cell-like phenotype to a neurite-bearing sympathetic neurone-like phenotype. Here we present data on the effect of infection of PC12 cells with retroviruses carrying the src oncogene of Rous sarcoma virus. Previous studies have demonstrated that the expression of src severely affects the synthesis and accumulation of differentiated cell products in a variety of cell types. We show that in the PC12 cell system, expression of v-src appears to have an inductive effect on differentiation that resembles the action of a 'physiological' growth factor.     

Patterning the neuronal cells via inkjet printing of self-assembled peptides on silk scaffolds

The patterning of neuronal cells and guiding neurite growth are important for neuron tissue engineering and cell-based biosensors. In this paper, inkjet printing has been employed to pattern self-assembled I₃QGK peptide nanofibers on silk substrates for guiding the growth of neuron-like PC12 cells. Atomic force microscopy (AFM) confirmed the dynamic self-assembly of I₃QGK into nanofiber structures. The printed self-assembled peptide strongly adheres to regenerated silk fibroin (RSF) substrates through charge-charge interactions. It was observed that in the absence of I₃QGK, PC12 cells exhibited poor attachment to RSF films, while for RSF surfaces coated or printed with peptide nanofibers, cellular attachment was significantly improved in terms of both cell density and morphology. AFM results revealed that peptide nanofibers can promote the generation of axons and terminal buttons of PC12 cells, indicating that I₃QGK nanofibers not only promote cellular attachment but also facilitate differentiation into neuronal phenotypes. Inkjet printing allows complex patterning of peptide nanofibers onto RSF substrates, which enabled us to engineer cell alignment and provide an opportunity to direct axonal development in vitro. The live/dead assay showed that printed I₃QGK patterns exhibit no cytotoxicity to PC12 cells demonstrating potential for future nerve tissue engineering applications.