运用基质辅助激光解吸电离飞行时间质谱技术建立鼻咽癌特异血清多肽谱诊断模型

目的 探讨鼻咽癌患者与正常健康者血清蛋白谱的差异,建立特异的鼻咽癌血清多肽谱诊断模型.方法 收集治疗前的鼻咽癌患者和正常健康对照者的血清标本,通过特异的液体磁珠分选后,进行基质辅助激光解吸电离飞行时间质谱技术(MALDI-TOF MS)分析,得到鼻咽癌患者的血清多肽谱.采用专用的生物信息学分析软件ClinProtTM进行差异分析,运用遗传算法,建立鼻咽癌的血清多肽谱诊断模型.通过盲法检验模型的灵敏度和特异度.结果 检测出鼻咽癌患者与正常健康者的差异蛋白峰有99个,选取两组间差异有统计学意义的、相对分子质量为808.99、834.61、3954.82和8141.88的蛋白峰,建立鼻咽癌诊断模型,该模型鼻咽癌的识别率为90.0%,预测能力为84.3%,盲法榆验模型的灵敏度为80.0%,特异度为64.0%.结论 鼻咽癌患者与正常对照者的血清间存在蛋白表达的差异,建立的特异的血清多肽谱诊断模型在鼻咽癌的诊断中有一定的应用前景,为寻找特异的鼻咽癌血清肿瘤标记物提供了一定的依据.

Abstract：

Objective To determine the specific serum peptide profile by comparing the serum differences between nasopharyngeal carcinoma patients (NPC) and normal control subjects, and to provide a diagnostic model of nasopharyngeal carcinoma. Methods Pre-treatment serum samples of NPC and normal control subjects were collected and assayed by MALDI-TOF MS analysis. The peptides were extracted with magnetic beads coated with WCX. Mass spectrographic data were analyzed with ClinProtTM software. The specific serum peptide model of NPC was established by using genetic algorithms. The sensitivity and specificity of model were tested by blind testing. Results The serum peptidome patterns of nasopharyngeal carcinoma was obtained. Differential expression of 99 peptide peaks was deteced, and the 808.99 Da,834.61 Da, 3954.82 Da, 8141.88 Da peptide peaks showing statistically significant differences between the two groups, were used to establish the diagnostic model for nasopharyngeal cancer. The recognition rate and predictive power of the model were 90.0% and 84.3%, respectively. The sensitivity and specificity of the model were 80. 0% and 64. 0% determined by blind testing, respectively. Conclusions Significant differences of serum peptide peaks are detected between NPC and normal control groups. The established specific serum peptide model may have certain application in the diagnosis of nasopharyngeal carcinoma, and provides the basis for discovering specific tumor markers of nasopharyngeal carcinoma.     

精子抗原肽P10G和YLP12的合成及在ELISA检测中的意义

目的 人工合成精子肽P10G和YLP12,以此为抗原建立检测抗精子抗体(AsAb)的酶联免疫吸附测定法(ELISA).方法 用PS3全自动合成仪合成精子肽P10G和YLP12,经反相高效液相色谱纯化和质谱鉴定.分别包板制备ELISA试剂盒,用以检测血清中的AsAb.结果 成功合成两种目的肽,质谱结果显示其相对分子质量均与理论相对分子质量相吻合,高效液谱法(HPLC)结果显示其纯度为97.5％和97.8％.以该两种合成肽为抗原,建立AsAb的ELISA,与商品化试剂盒比较均显示极好的一致性(Kappa值为0.77和0.79,＞0.75).结论 全自动固相多肽合成技术可用于合成高纯度的精子抗原肽,所合成的P10G和YLP12均具有较好的抗原活性,可作为包被抗原用于AsAb的ELISA检测.     

Surface modification by cold-plasma technique for dental implants—Bio-functionalization with binding pharmaceuticals

Since biomaterials contact many different tissues, those materials must have optimum surface compatibility with the host bone tissue and soft tissue, as well as anti-microbial properties on an exposed region of the mucosa. Such materials can be created under well-controlled conditions by modifying the surfaces of materials that contact those tissues. This paper is focused on the surface modification of biomaterials for developing “Bio-functional dental implants”, which are compatible with all host tissues, using a cold-plasma technique.At the bone tissue/implant interface, a thin calcium phosphate coating and rapid heating with infrared radiation were effective in controlling the dissolution without cracking the coating. These thin calcium phosphate coatings may directly promote osteogenisis, but also enable immobilization and subsequent drug delivery system (DDS) of bisphosphonates. Simvastatin is also an effective candidate that is reported to increase the expression of BMP-2. The thin-film of hexamethyldisiloxane (HMDSO) was plasma-polymerized onto titanium, and then HMDSO surface was activated by O₂-plasma treatment. A quartz crystal microbalance (QCM-D) technique demonstrated that simvastatin was immobilized on the plasma-treated surfaces due to introduction of O₂-functional groups. At the soft tissue/implant interface, multi-grooved surface topographies and utilizing the adhesive proteins such as fibronectin or laminin-5 may help in providing a biological seal around the implant. At the oral fluid/implant interface, an alumina coating, F⁺-implantation and immobilization of anti-microbial peptides were responsible for inhibiting the biofilm accumulation.     

阻断钙调磷酸酶/激活T细胞核因子通路对聚甲基丙烯酸甲酯颗粒抑制骨祖细胞向成骨细胞分化的影响

目的 探讨VIVIT肽阻断钙调磷酸酶(Cn)/激活T细胞核因子(NFAT)信号通路对聚甲基丙烯酸甲酯(PMMA)颗粒抑制骨祖细胞向成骨细胞分化的影响. 方法 体外分离培养Sprague-Drawley大鼠胎鼠颅骨原代细胞(包含大量骨祖细胞),根据处理条件不同分为4组:对照组、PMMA组、PMMA/VIVIT组和VIVIT组.细胞培养2、4、7和14 d用MTT法检测细胞增殖情况,7 d和14d用碱性磷酸酶(ALP)定量反映细胞分化;细胞培养14 d后菏素红染色观察细胞矿化,RT-PCR法观察ALP、骨钙素、Ⅰ型胶原、Fra-2(与成骨细胞分化有关的转录因子)、NFATc1的基因表达,Western Blot法检测细胞核和细胞质中NFATc1蛋白的表达. 结果 PMMA组较对照组NFATc1基因和蛋白表达增加,并伴有NFATc1蛋白转位入核明显增加,成骨细胞分化、矿化和相关基因表达明显降低.PMMA/VIVIT组较PMMA组细胞分化、矿化和相关基因表达增加,但低于VIVIT组,NFATc1基因表达降低,转位入核的NFATc1蛋白明显减少.各组细胞增殖差异均无统计学意义(P>0.05). 结论 PMMA颗粒抑制骨祖细胞向成骨细胞分化与Cn/NFAT信号通路激活有关,VIVIT肽阻断Cn/NFAT信号通路可促进PMMA颗粒抑制的骨祖细胞向成骨细胞分化.     

The outcome of cross‐priming during virus infection is not directly linked to the ability of the antigen to be cross‐presented

The initiation of CD8⁺ T cell (CTL) immune responses can occur via cross‐priming. Recent data suggested a relationship between cross‐presentation and immunodominance of epitope‐specific T cells. To test this association, we evaluated the efficacy of cross‐presentation for several virus epitopes in vitro and examined if this can be extrapolated in vivo. Employing lymphocytic choriomeningitis virus (LCMV), we demonstrate that the cross‐presentation and cross‐priming of LCMV antigens were dominated by NP396, but not NP205 when analyzing the LCMV‐NP. Although with LCMV‐GP, cross‐presentation was dominated by GP276, and cross‐priming was dominated by GP33. Importantly, although NP396 was significantly more efficient than GP33 in cross‐presentation, cross‐priming of their specific CTL was comparable. In a subsequent virus challenge after cross‐priming, GP33‐specific CTL dominated the response. Accordingly, based on our data, the ability of viral epitopes to be cross‐presented in vitro does not entirely reflect what would occur in cross‐priming. Thus, weak cross‐presenting antigens may still cross‐prime an efficient CTL response depending on other in vivo elements such as the naïve T‐cell precursor frequencies.     

海洋胶原肽的分子组成及其降血脂和抗氧化作用研究

目的 测定海洋胶原肽(marine collagen peptide,MCP)的相对分子质量,观察其对实验性高脂血症大鼠的血脂水平和抗氧化酶活性及脂质过氧化产物丙二醛(MDA)含量的影响.方法 采用葡聚糖凝胶层析、高效液相和基质辅助激光解吸电离飞行时间质谱方法对MCP组分进行分离和相对分子质量测定.选用50只健康雄性SD大鼠分成正常对照、高脂模型对照和1.0、3.0、9.0 g/kg MCP组,正常对照组大鼠喂饲普通饲料,高脂模型对照组和MCP组大鼠喂饲高脂饲料(79%基础饲料+10%猪油+10%蛋黄粉+1%胆固醇),MCP组大鼠以灌胃方式连续给予不同剂量的MCP(2 ml/100 g)45 d,测定血脂水平及血清谷胱甘肽过氧化物酶(GSH-Px)活力、超氧化物歧化酶(SOD)活性和MDA含量.结果 MCP的相对分子质量范围为100～860,摄人MCP 45 d后1.0、3.0和9.0 g/kg剂量组大鼠血清总胆固醇(TC)分别为(1.89±0.29)mmol/L、(2.07±0.39)mmol/L和(1.99±0.29)mmol/L,与高脂模型对照组(3.37±0.24)mmol/L相比明显降低;血清低密度脂蛋白胆固醇(LDL-C)水平分别为(0.83±0.16)mmol/L、(1.01±0.35)mmol/L和(0.91±0.26)mmol/L,明显低于高脂模型对照组(2.20±0.34)mmol/L;血清SOD活性分别为(218.6±33.2)U/ml、(242.7±21.4)U/ml和(242.1±44.8)U/ml,与高脂模型对照组(119.7±47.8)U/ml相比有显著升高;动脉粥样硬化指数(AI)分别为1.14±0.22、1.16±0.27和0.99±0.31,明显低于高脂模型对照组(2.27±0.55);抗动脉粥样硬化指数(AAI)分别为0.47±0.04、0.47±0.06和0.51±0.09,与高脂模型对照组(0.31±0.05)相比显著增高;3.0和9.0 g/kg剂量组大鼠血清甘油三酯(TG)水平分别为(0.90±0.15)和(0.86±0.12)mmol/L,明显低于高脂模型对照组(1.18±0.18)mmol/L;9.0g/kgbw MCP组大鼠血清MDA含量(7.1±4.1)nmol/ml明显低于高脂模型对照组(15.9±9.9)nmol/ml.结论 MCP具有降血脂和抗氧化的生物活性,对高脂血症的形成和动脉粥样硬化的发生具有一定的预防作用.     

Reports on Dietary Intervention in Autistic Disorders

Autism is a developmental disorder for which no cure currently exists. Gluten and/or casein free diet has been implemented to reduce autistic behaviour, in addition to special education, since early in the eighties. Over the last twelve years various studies on this dietary intervention have been published in addition to anecdotal, parental reports. The scientific studies include both groups of participants as well as single cases, and beneficial results are reported in all, but one study. While some studies are based on urinary peptide abnormalities, others are not. The reported results are, however, more or less identical; reduction of autistic behaviour, increased social and communicative skills, and reappearance of autistic traits after the diet has been broken.