云芝糖肽的结构组成分析

目的:对云芝糖肽的结构与组成进行研究.方法:采用还原衍生化气相色谱,紫外、核磁、高碘酸氧化、Smith降解、琼脂糖凝胶电泳等方法.结果:云芝糖肽由葡萄糖、半乳糖、岩藻糖、甘露糖、木糖和鼠李糖组成,具有α,β型糖苷键,连接方式有(1→4)、(1→6)两种,含有结合态蛋白,糖肽键类型初步确定为N-连接.结论:首次对云芝糖肽的化学结构与组成进行了较为细致的研究.     

Immunization with glycosylated Kb-binding peptides generates carbohydrate-specific,unrestricted cytotoxic T cells

Cytotoxic T cells (CTL) recognize target proteins as short peptides presented by major histocompatibility complex (MHC) class I restriction elements. However, there is also evidence for peptide-independent T cell receptor (TCR) recognition of target proteins and non-protein structures. How such T cell responses are generated is presently unclear. We generated carbohydrate (CHO)-specific, MHC-unrestricted CTL responses by coupling di- and trisaccharides to K^b- or D^b-binding peptides for direct immunization in mice. Four peptides and three CHO have been analyzed with the CHO either in terminal or central positions on the carrier peptide. With two of these glycopeptides, with galabiose (Galα1-4Gal; Gal₂) bound to a homocysteine (via an ethylene spacer arm) in position 4 or 6 in a vesicular stomatitis virus nucleoprotein-derived peptide (RGYVYQGL binding to K^b), CTL were generated which preferentially killed target cells treated with glycopeptide compared to those treated with the core peptide. Polyclonal CTL were also found to kill target cells expressing the same Gal₂ epitope in a glycolipid. By fractionation of CTL, preliminary data indicate that glycopeptide-specific K^b-restricted CTL and unrestricted CHO-specific CTL belong to different T cell populations with regard to TCR expression. The results demonstrate that hapten-specific unrestricted CTL responses can be generated with MHC class I-binding carrier peptides. Different models that might explain the generation of such responses are discussed.     

薄芝糖肽注射液治疗儿童急性面神经炎疗效观察

目的：观察薄芝糖肽注射液治疗儿童急性面神经炎的疗效。方法将50例患儿随机分为对照组、实验组，对照组22例给予常规治疗，实验组26例在常规治疗基础上加用薄芝糖肽注射液治疗。2个疗程后观察2组临床疗效。结果对照组治愈7例（31.8％），好转8例（36.4％），无效7例（31.8％）；实验组治愈16例（61.5％），好转9例（34.6％），无效1例（3.8％）；实验组治愈率和总有效率均高于对照组，差异有统计学意义（ P＜0.05）。结论薄芝糖肽注射液能明显提高急性面神经炎患儿的治愈率和总有效率，值得临床应用。     

Targeted proteomic strategy for clinical biomarker discovery

The high complexity and large dynamic range of blood plasma proteins currently prohibit the sensitive and high‐throughput profiling of disease and control plasma proteome sample sets large enough to reliably detect disease indicating differences. To circumvent these technological limitations we describe here a new two‐stage strategy for the mass spectrometry (MS) assisted discovery, verification and validation of disease biomarkers. In an initial discovery phase N‐linked glycoproteins with distinguishable expression patterns in primary normal and diseased tissue are detected and identified. In the second step the proteins identified in the initial phase are subjected to targeted MS analysis in plasma samples, using the highly sensitive and specific selected reaction monitoring (SRM) technology. Since glycosylated proteins, such as those secreted or shed from the cell surface are likely to reside and persist in blood, the two‐stage strategy is focused on the quantification of tissue derived glycoproteins in plasma. The focus on the N‐glycoproteome not only reduces the complexity of the analytes, but also targets an information‐rich subproteome which is relevant for remote sensing of diseases in the plasma. The N‐glycoprotein based biomarker discovery and validation workflow reviewed here allows for the robust identification of protein candidate panels that can finally be selectively monitored in the blood plasma at high sensitivity in a reliable, non‐invasive and quantitative fashion.     

溶血葡萄球菌对抗菌药物的敏感性和mecA基因的检测

目的　了解溶血葡萄球菌的耐药情况 ,以指导临床合理用药。方法　对 133株临床分离的溶血葡萄球菌采用微量肉汤稀释法测定 16种抗菌药物的最低抑菌浓度 (MIC) ,多聚酶链反应 (PCR)法检测mecA基因 ,并与普通药敏试验比较 ,对不相符的菌株进行抑制、诱导试验。结果　 133株溶血葡萄球菌对利奈唑胺、万古霉素、去甲万古霉素无耐药 ,对替考拉宁的耐药率为 6 .0 % ;对阿米卡星、异帕米星、米诺环素、利福平、氟氧头孢的耐药性较低 ,耐药率 <2 0 % ;对青霉素、苯唑西林的耐药率 >90 %。除米诺环素、青霉素外 ,住院患者分离菌株的耐药率高于门诊患者分离的菌株。mecA基因阳性率为 90 .2 3% ,且PCR产物经克隆测序证实为mecA特异性产物。对 2株mecA基因阳性而苯唑西林MIC≤ 0 .2 5 μg/ml的菌株进行诱导试验后其MIC≥ 0 .5 μg/ml;对 3株mecA基因阴性而MIC≥ 0 .5 μg/ml的菌株进行抑制试验后其MIC值下降 8倍以上。 结论　溶血葡萄球菌对大多数抗菌药物均有较高的耐药性 ,对糖肽类抗生素的敏感性也在下降 ,需引起临床重视。     

关节镜清理联合静脉抗生素治疗膝关节革兰阳性菌感染的疗效观察

目的探究关节镜清理、关节腔灌洗联合静脉应用糖肽类抗菌药物治疗革兰阳性菌所致膝关节感染的疗效。方法 2008年2月-2010年7月应用关节镜清理、关节腔灌洗联合静脉糖肽类抗菌药物治疗革兰阳性菌所致膝关节感染患者21例,其中耐甲氧西林金黄色葡萄球菌（MRSA）感染5例,耐甲氧西林表皮葡萄球菌（MRSE）感染6例,肠球菌感染4例,凝固酶阴性葡萄球菌（CONS）感染6例,药敏试验显示培养菌群均对糖肽类抗菌药物万古霉素及替考拉宁敏感,观察药物治疗过程中不良反应情况,评价临床疗效。结果治疗期间未发现药物过敏反应和其他与药物有关的不良反应记录,治疗有效率为100%。本组患者均获得随访,时间4-29个月,平均17.6个月,随访期间患者均无复发感染症状。结论关节镜清理、关节腔灌洗联合静脉应用糖肽类抗菌药物是治疗膝关节革兰阳性菌感染的一种有效方法。     

Glycopeptide susceptibility profiles of nosocomial multiresistant Staphylococcus haemolyticus isolates

We investigated 48 Staphylococcus haemolyticus isolates from patients and medical staff in terms of susceptibility to and in-vitro selection for vancomycin and teicoplanin in regard to their antibiotypes. On comparison of multiresistant S. haemolyticus isolates with non-multiresistant isolates, the geometric mean minimum inhibitory concentration (MIC) of vancomycin for multiresistant S. haemolyticus was 2.9 μg/ml, and that of teicoplanin was 18.0 μg/ml, both of which values were significantly greater than the corresponding mean MICs of vancomycin (2.0 μg/ml) and teicoplanin (4.7 μg/ml) for nonmultiresistant isolates. After agar selection, the mean of the highest teicoplanin concentration of selected plates for multiresistant S. haemolyticus was 97.1 μg/ml, which was significantly higher than that for nonmultiresistant isolates (57.8 μg/ml). However, the means' of the highest vancomycin concentrations after agar selection for multiresistant and nonmulti-resistant isolates were the same, at 7.4 μg/ml, with no colonies capable of growing in 32 μg/ml of vancomycin. There was no significant difference in glycopeptide susceptibility between oxacillin-resistant and oxacillin-susceptible isolates among nonmultiresistant S. haemolyticus. The geometric mean MICs of vancomycin for oxacillin-resistant and oxacillin-susceptible isolates were 2.1 μg/ml and 1.6 μg/ml, and those of teicoplanin were 4.4 μg/ml and 5.6 μg/ml, while the means of the highest concentrations of the selected plates of vancomycin were 8.6 μg/ml and 3.3 μg/ml, and those of teicoplanin were 52.8 μg/ml and 74.7 μg/ml, respectively. Multiresistant isolates showed significantly greater mean MICs of vancomycin and teicoplanin and higher teicoplanin concentration of the selected plates than nonmultiresistant isolates, irrespective of oxacillin resistance. These results indicate that methicillin resistance may not be related to reduced susceptibility to glycopeptide in S. haemolyticus, and that a multiresistant profile is associated more with a decreasing susceptibility to glycopeptides then with resistance to oxacillin. In this study, antibiotypes showed good concordance with pulsed-field gel electrophoresis typing results, with a sufficiently high discriminatory ability index, of 0.912. We consider that primary screening with antimicrobial susceptibility testing and antibiotyping, with attention to the multiresistant profile, would be useful for monitoring nosocomial S. haemolyticus colonization and infection. Received: August 9, 2000 / Accepted: February 5, 2001     

猪红细胞膜糖肽抗肿瘤作用的初步研究

目的:探讨猪红细胞膜糖肽的抗肿瘤转移作用;方法:通过腹腔注射给药测定其对小鼠肿瘤生长和转移的影响。结果:糖肽明显抑制S_(180)肿瘤及Lewis肺癌的生长,抑制率分别为52.62％和66.44％,对Lewis肺癌的自发性肺转移的抑制率为78.16％。结论:猪红细胞膜糖肽具有抗肿瘤生长及抗转移的生物学活性,是一很有发展前景的生化制剂。