Syntheses and Functional Studies of Self‐Adjuvanting Anti‐HER2 Cancer Vaccines

The 9‐mer peptide MFCH401 (N: 165–173: DTILWKDIF), which is located in the extracellular domain of HER2, has been predicted to be a novel epitope. Self‐adjuvanting anti‐HER2 vaccine constructs were designed and synthesized via covalently attaching MFCH401 or its linear tandem repeats (2×MFCH401, 3×MFCH401) to a lipopeptide Pam₃CSK₄ via iterative condensation reaction. The in vivo results showed the Pam₃CSK₄‐MFCH401 vaccine construct can induce higher antibody titers of IgG and IgM than those of other conjugates, and the analysis of changes in plasma cytokines level indicate the activation of Th1 cells and NK cells. In addition, the Pam₃CSK₄‐MFCH401 vaccine conjugate induced a specific immune response to HER2‐overexpressing human BT474 cells. Our data clearly indicated that MFCH401 is a promising epitope; moreover, its linear tandem repeats were unsuitable for anticancer vaccine design when conjugating with Pam₃CSK₄, which provided useful evidence for developing further anti‐HER2 cancer vaccines.     

Analysis and dynamical behavior of fractional-order cancer model with vaccine strategy

In recent year, the world has witnessed the arrival of deadly diseases like cancer over all the global levels. To fight back this disease or control the spread, mankind relies on modeling and medicine to control, cure, and behavior of the cancer diseases. We developed the fractional-order immunotherapy bladder cancer model and used the BCG vaccine for treatment by using the Caputo fractional derivative operator φɛ(0,1] . A mathematical model has four variables B,E, T_i, T_u which represent the vaccine for the immune system, effector cells, total population of affected, and unaffected cells, respectively. In this model, we have two cases according to the growth rate of cells. The fractional-order system is stable in both cases and gives the solution infeasible region for uniqueness, positivity, and boundedness to illustrate the treatment of cancer. The effect of fractional parameter on our obtained solutions is presented, and a comparison is made with the classical ordinary derivative operator. It is worthy to observe that fractional derivatives show significant changes and memory effects as compared with ordinary derivatives to control the disease at the initial stage to overcome the risk of living with cancer.     

Four-step approach to efficiently develop capillary gel electrophoresis methods for viral vaccine protein analysis

Vaccines against infectious diseases are urgently needed. Therefore, modern analytical method development should be as efficient as possible to speed up vaccine development. The objectives of the study were to identify critical method parameters (CMPs) and to establish a set of steps to efficiently develop and validate a CE-SDS method for vaccine protein analysis based on a commercially available gel buffer. The CMPs were obtained from reviewing the literature and testing the effects of gel buffer dilution. A four-step approach, including two multivariate DoE (design of experiments) steps, was proposed, based on CMPs and was verified by CE-SDS method development for: (i) the determination of influenza group 1 mini-hemagglutinin glycoprotein; and (ii) the determination of polio virus particle proteins from an inactivated polio vaccine (IPV). The CMPs for sample preparation were incubation temperature(s) and time(s), pH, and reagent(s) concentration(s), and the detection wavelength. The effects of gel buffer dilution revealed the CMPs for CE-SDS separation to be the effective length, the gel buffer concentration, and the capillary temperature. The four-step approach based on the CMPs was efficient for the development of the two CE methods. A four-step approach to efficiently develop capillary gel electrophoresis methods for viral vaccine protein analysis was successfully established.     

壳聚糖及其衍生物作为疫苗佐剂的研究进展

疫苗佐剂能够增强机体对抗原的免疫应答反应或改变免疫应答反应类型,延长疫苗在体内作用时间,提高疫苗效力。壳聚糖能有效地将疫苗递送到靶抗原递呈细胞或组织,激活抗原提呈细胞,诱导产生免疫应答,促进Th1/Th2应答反应的平衡,因此,壳聚糖作为疫苗佐剂具有一定的潜力。为了解决壳聚糖在中性和碱性溶液中溶解性差,以及进一步提高其黏膜黏附性和靶向性等问题,通过对壳聚糖进行化学改性,生成一系列壳聚糖衍生物,提高其佐剂性能。本论文就近年来有关壳聚糖及其衍生物作为疫苗佐剂和递送系统在疫苗中的应用进行了综述,总结并提出了壳聚糖及其衍生物在疫苗佐剂应用领域所面临的问题以及其未来的发展方向,使读者对其有全面的了解。     

HPAEC‐PAD method for the analysis of alkaline hydrolyzates of Haemophilus influenzae type b capsular polysaccharide

A gradient method has been devised for the rapid analysis of alkaline hydrolyzates of Haemophilus influenzae type b (Hib) capsular polysaccharide‐based vaccines by high‐performance anion exchange chromatography with pulsed amperometric detection (HPAEC‐PAD). As compared with published procedures, peak shape and sensitivity were significantly improved with this approach, analysis time was short and there was little interference from impurities. The limits of detection and quantification were established with a purified reference polysaccharide. We propose this method as a practical alternative for the analysis of minute amounts of Hib polysaccharide, which can be lower than with the conventional approaches. Copyright © 2013 John Wiley & Sons, Ltd.     

Urinary metabolite profiling provides potential differentiation to explore the mechanisms of adjuvant‐induced arthritis in rats

To explore the pathogenesis of rheumatoid arthritis (RA) from the perspective of metabolomics, gas chromatography time‐of‐flight mass spectrometry (GC‐TOF/MS) technology was used to observe changes in the metabolic profiles of urine output from rats with adjuvant‐induced arthritis (AA). Sprague–Dawley rats were randomly divided into a control group and an experimental group, with eight in each. Rats in the experimental group were induced by intracutaneous innoculation of 0.1 mL Freund's complete adjuvant to right paws. On day 20 after immunization, the metabolic profiles between rat control and experimental groups were compared by combining GC‐TOF/MS technology with multivariate statistical approaches, including principal component analysis, partial least squares discriminant analysis and orthogonal projections to latent structures–discriminant analysis. Nine potential biomarkers were identified, including 2,2‐dimethylsuccinic acid, tartronic acid, dehydroshikimic acid, hippuric acid, adenine, phenaceturic acid, l ‐dopa, 1,4‐dihydroxy‐2‐naphthoic acid and melibiose. The findings indicate that the rats with AA are disturbed in metabolism of purine, amino acid, fat and energy. This study also demonstrates that the dysfunction in a range of biosynthetic and catabolic pathways, which leads to increased oxygen free radicals and inflammation, could cause underlying pathogenesis of RA. Copyright © 2016 John Wiley & Sons, Ltd.     

高效液相色谱法测定黑色素瘤小鼠尿液中达卡巴嗪

达卡巴嗪是治疗恶性黑色素瘤的一线化疗药物。它在体内主要经肝脏代谢,同时部分药物仍以原药形式经尿液排出。这就意味着可以通过监测尿液中达卡巴嗪的含量评估其在人体内的利用率和转化率,进而对其治疗效果进行评价。针对达卡巴嗪,人们发展了多种分析方法,但多基于高效液相色谱-质谱平台。然而达卡巴嗪为强极性弱碱性化合物,采用常规反相色谱法分析时会出现出峰时间过早、峰形拖尾的现象,导致定量不准确。基于此,该文建立了一种测定尿液中达卡巴嗪含量的高效液相色谱方法以克服上述问题。小鼠尿液经丙酮沉淀法去除蛋白后,采用Shimadzu-GL ODS柱（150 mm×4.6 mm,5 μm）分离,色谱条件如下：流动相为甲醇/乙腈（1：1,v/v）-0.01 mol/L磷酸二氢钠溶液（pH=6.5）（20：80,v/v）,流速1 mL/min,检测波长280 nm,柱温35℃,洗脱时间15 min。在该色谱条件下,达卡巴嗪保留时间为5.3 min且峰形良好。其在0.25~1000 μg/mL范围内线性关系良好（r ² =0.999）。基于信噪比（S /N ）=3和S /N =10,计算出检出限和定量限分别为0.12 μg/mL和0.25 μg/mL。在低、中、高（50.0、375、500 μg/mL）3个添加水平下,加标回收率分别为98.9%、102%、99.1%,相对标准偏差（RSD）分别为3.2%、1.3%、1.2%（n =5）。日内与日间RSD分别小于3.8%和4.4%。将该法应用于不同发展阶段的黑色素瘤C57BL/6小鼠尿液中达卡巴嗪的监测,结果表明该方法操作简便,结果可靠,便于推广。