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.     

Presence of hepcidin-25 in biological fluids: Bile,ascitic and pleural fluids

AIM: To examine body fluids such as ascitic fluid (AF),saliva,bile and pleural effusions for the presence of hepcidin using a novel radioimmunoassay (RIA).METHODS: Serum samples were collected from 25 healthy volunteers (mean age: 36 ± 11.9 years,11 males,14 females).In addition bile was obtained from 12 patients undergoing endoscopic retrograde cholangiopancreatography (mean age: 66.9 ± 16.7 years,M:F = 5:7).Saliva was collected from 17 healthy volunteers (mean age: 35 ± 9.9 years,M:F = 8:9).Pleural and AF...     

葵花粕中绿原酸的提取及其抗氧化研究

采用乙醇回流提取法提取葵花粕中的绿原酸,探讨乙醇溶液体积分数、料液比、提取温度、提取时间、pH值对绿原酸提取量的影响,采用正交试验对绿原酸的提取工艺进行优化,并对葵花粕绿原酸的抗氧化活性进行了研究。结果表明:绿原酸的最佳提取工艺条件为乙醇溶液体积分数55%、提取温度60℃、提取时间1.5h、料液比1∶12(g.mL-1)、pH值6、提取1次。     

4种促性腺激素释放激素抗体的制备、鉴定和纯化

用3-马来酰亚胺基苯甲酸琥珀酰亚胺酯(MBS)法将人工合成的4种不同来源的促性腺激素释放激(GnRH)(即章鱼GnRH、海鞘GnRH-Ⅰ、七鳃鳗GnRH-Ⅰ和七鳃鳗GnRH-Ⅲ)分别与匙孔戚血蓝蛋白(KLH)偶联后作为抗原,免疫新西兰大白兔获得GnRH抗体.用间接ELISA方法检测抗血清效价,并用Western blot鉴定其特异性.制备抗原肽亲和纯化柱,纯化多克隆抗体.结果显示,4种多克隆抗体效价高达1∶500 000(体积比),并能和相应的GnRH多肽特异性结合,纯化后的抗体在SDS-PAGE显示为单一条带.通过上述方法,获得4种高效价、高纯度的GnRH抗体,为低等动物GnRH的研究提供了理论依据.     

使用伪氨基酸组成和BP神经网络预测类弹性蛋白多肽的相变温度

根据获得的16条ELP序列及相变温度的数据,利用伪氨基酸组成方法提取其序列特征值.将伪氨基酸组成中的相关系数部分作为类弹性蛋白的特征向量,从类弹性蛋白序列出发,利用最小中位方差回归,找出与其序列相关系数的最佳阶数.运用均匀设计法,分别对支持向量机与BP神经网络参数进行优化.结果表明:BP神经网络获得的预测模型最佳,相变...     

铁离子还原法(FRAP)测定中国被毛孢菌丝体提取物抗氧化活性

FRAP法测定了6批人工培养中国被毛孢(Hirsutella sinensis)菌丝体热水和乙醇提取物的抗氧化活性.结果表明:1 mg/mL人工培养中国被毛孢丝体水提物和乙醇提取物的平均抗氧化活性分别相当于(327.5±42.1)μmol/mL Fe2+和(107.1±13.5)μmol/mL Fe2+;方法学研究显示重现性为3.8%;精密度为3.1%;可靠性相关系数R2为1.0000;回收率分别为(105.4±5.8)%(水提物)和(104.2±8.8)%(醇提物);该方法在30 min内结果的变异系数为3.0%.这些结果表明:中国被毛孢菌丝体热水和乙醇提取物均具有一定的抗氧化活性,用FRAP法测定具有良好的重现性、精密度、可靠性、回收率和稳定性.     

Application of surface plasmon resonance in screening adsorbents and explaining adsorption phenomena using model polymers

A surface plasmon resonance (SPR) technique was developed as an online screening method to identify effective adsorbents for the peptide NH₂-Cys-Thr-Trp-Trp-COOH (CW-4). To validate the SPR technique, several linear polymers containing amino acid residues were synthesized as models of their corresponding adsorbents. SPR screening of all the linear polymers demonstrated that the linear polymer containing phenylalanine residues (PPhe) exhibited the highest affinity for CW-4 among the polymers tested. In accordance with the screening results from SPR, the adsorbent containing phenylalanine (APhe) exhibited the highest adsorption capacity for CW-4. An interesting observation was that the amounts of CW-4 adsorbed on 3 adsorbents were quite small, even though both SPR and isothermal titration calorimetry (ITC) experiments indicated the presence of interactions between CW-4 and the three polymers. A kinetic analysis performed using SPR technology suggested such a phenomenon was ascribed to the poor stability of the CW-4/polymer complexes; that is, CW-4 associates with the polymers quickly, but it also dissociated quickly. The combined results suggested that SPR was a promising tool to identify the optimal adsorbent for peptides, analyze the interactions contributing to adsorption and explain adsorption phenomena using polymer models.     

胃癌高发区不同人群血浆总抗氧化力及谷脱甘肽过氧化物酶活性的研究

用微弱化学发光法和酶联分光光度技术研究胃癌高发区－－甘肃省武威地区不同人血浆总抗氧化力（TRAC）和谷胱甘肽过氧化物酶（GSH－Px）活性，结果表明：中度表浅性胃炎18例、21例明显降低(p-≤0．01），但各组间血浆GSH－Px活性无显著性差异（P＞0．05），提示当地胃癌高发可能与血产供销 TRAC水平的降低有关，而与GSH－Px活性改变无明显相关。     

水解度对蚕蛹蛋白酶解物的物化特性和抗氧化能力的影响

对蛋白质进行适度酶解是改善动植物蛋白原料的物化性质和提高其生理活性的常用方法.研究了水解度对蚕蛹蛋白的溶解性、乳化性、起泡性等物化性质的影响,以及水解度对蚕蛹蛋白的还原力,对DPPH自由基、超氧阴离子自由基和羟基自由基的清除能力等抗氧化特性的影响.实验结果表明,蚕蛹蛋白经适度酶解后溶解性得到显著改善,且水解度越大溶解性越好.蚕蛹蛋白经酶解后乳化性与乳化稳定性均降低,但起泡性和起泡稳定性增强,水解度增至5%时,起泡性和起泡稳定性达到最高值,继续增大水解度,反而下降.蚕蛹蛋白经酶解后抗氧化性显著提高,水解度为5%时,还原力和对DPPH自由基和羟基自由基的清除能力最高,水解度为15%时,对超氧阴离子自由基的清除能力最高,继续增加水解度,抗氧化能力反而下降.这说明,对蛋白质原料进行酶解改性时,不能简单的追求高水解度,而应根据产品的要求控制合适的水解度,过度水解是无益的.     

文蛤蛋白水解制备抗氧化活性肽的研究

以文蛤蛋白为原料,通过蛋白酶水解制备抗氧化活性肽.利用体外清除DPPH活性评价酶解产物的抗氧化活性,采用超滤、葡聚糖凝胶柱层析和HPLC对相应的活性肽进行分离纯化,通过HPLC-MS进行结构鉴定.结果表明:木瓜蛋白酶水解产物的DPPH清除活性明显高于碱性蛋白酶、风味蛋白酶、中性蛋白酶和复合蛋白酶,产物质量浓度为1 mg·mL-1时,DPPH清除率为55.74%;以DPPH清除率为筛选指标,对木瓜蛋白酶酶解产物进行逐级分离,得到4个具有抗氧化活性的目的肽段,氨基酸序列分别为LNFNLEKSR(1120.3 u)、SWLRPR(814.4 u)、RIGNIISQY(1063.3 u)和LNFNLEK(877.2 u).