人格拉利素新抗菌肽的抗菌作用研究

目的:进一步证实人格拉利素(hGlyrichin)抗菌肽的抗菌功能.方法:根据人格拉利素的氨基酸序列,设计并合成了19个氨基酸长度的多肽(pCM-19),采用试管法和菌落生成法分析该肽对靶细菌生长的影响.结果与结论:pCM-19溶液与不同靶细菌共孵育后,明显抑制了细菌的生长,对实验室常用的BL-21工程菌和多种致病菌(包括杆菌、球菌)的生长都有抑制作用.更为有趣的是对耐甲氧西林金黄色葡萄球菌的生长有明显抑制作用,显示了其在抗耐药菌感染中的良好前景.     

新基因LRP15的功能预测及其编码蛋白定位

目的：确定白血病复发相关新基因LRP15编码蛋白的亚细胞定位，利用生物信息学探索网上预测基因功能的新方法。方法：以人类基因组资源(HGR)数据库为基础，进行LRPl5启动子区特征分析。在Prosite数据库中进行有生物学意义的保守性氨基酸修饰位点搜索。通过RPS-BLAST程序预测其蛋白质结构及功能。以增强绿色荧光蛋白(EGFP)为报告基因，利用激光共聚焦显微镜等实验方法验证生物信息学的分析结果。结果：LRP15编码蛋白含有cAMP依赖的蛋白激酶磷酸化位点及酪蛋白激酶Ⅱ磷酸化位点，其N端有一个富含亮氨酸重复序列，定位于细胞核内。结论：生物信息学是预测基因功能的有效方法；利用EGFP可以将LRPl5蛋白定位于细胞核内；LRPl5基因可能是一个参与细胞发育调控的新基因。     

99Tcm标记血管活性肠肽衍生物的制备及其生物分布研究

目的观察99Tcm标记血管活性肠肽(VIP)衍生物VIPSN3在生物体内的分布和肿瘤摄取情况。方法通过化学合成法合成含SN3类配体的VIPSN3。在VIP的羧基端连接4个氨基酸(其中1个含巯基),以形成含有SN3的四齿状结构作为99Tcm的强螯合基团,即VIPSN3。引入γ氨基丁酸(Aba)作为隔离物以消除空间阻碍。采用配体交换法进行99Tcm标记,测定99TcmVIPSN3的生物活性和功能、体外稳定性及其在荷结肠癌裸鼠的体内分布,并行γ显像。结果VIPSN3纯度经HPLC分析证实>99%。99TcmVIPSN3标记率为(90±8)%,体外活性分析证实其活性损失小,标记物体外稳定。动物实验示99TcmVIPSN3从血液清除快速,经肾、膀胱排除。荷结肠癌裸鼠肿瘤在注射99TcmVIPSN3后24h显像清晰,除肾外,肿瘤摄取最高,与99TcmVIPN4相比,肾脏摄取明显减少。体内分布结果示99TcmVIPSN3注射后24h,肿瘤每克组织百分注射剂量率(%IDg)为058±015,比99TcmVIPN4(026±001)高,瘤血比值>2,瘤肌肉比值>10。结论VIPSN3具有天然VIP的生物活性,99TcmVIPSN3肾脏摄取较低,肿瘤显像清晰,是一个较有希望的肿瘤显像剂。     

低分子量尿激酶与血纤蛋白粘附肽(12肽)融合表达及特性分析

人工合成血纤蛋白粘附肽（１２肽）ｃＤＮＡ，与低分子量单链尿激酶基因重组，获得血纤蛋白粘附肽与低分子量单链尿激酶的融合基因。将此融合基因重组入ｐＢＶ２２０表达载体并在大肠杆菌中表达，表达产物具有与天然尿激酶相同的抗原性和溶纤活性，同时兼具血纤蛋白粘附肽（１２肽）的抗血纤蛋白单体聚合的活性     

介导感觉生理功能的瞬时受体电位通道蛋白(TRP)家族研究进展

瞬时受体电位通道蛋白(TRP)家族由一类特殊的阳离子通道蛋白组成,在神经细胞及其他非兴奋细胞中有重要作用,其中在介导多种感觉生理功能方面的作用尤其显著.TRP结构与功能的深入研究为阐明感觉生理功能的分子机制提供了重要线索.本文综述TRP家族在温度感受、机械刺激感受、光感受和化学信号感受等方面的研究进展.     

Imaging of infection in rabbits with radioiodinated interleukin-1 (α and β), its receptor antagonist and a chemotactic peptide: a comparative study

Previous studies have reported the favourable characteristics of chemotactic peptides and interleukins for imaging of infection and inflammation. In the present study, the potential of two species of interleukin 1 (IL-1), IL-1α and IL-1β, the IL-1 receptor antagonist (IL-1ra) and the synthetic chemotactic peptide N-formyl-methionyl-leucyl-phenylalanyl-lysine (fMLFK) were directly compared in a rabbit model of infection. IL-1α, IL-1β, IL-1ra and fMLFK were labelled with iodine-123 according to the Bolton-Hunter method. Twenty-four hours after induction of Escherichia coli abscesses in the left thigh muscle, rabbits were injected intravenously with 0.5 mCi of ¹²³I-labelled agent. Gamma camera images were obtained at 5 min and 1, 4, 8 and 20 h p.i. Biodistribution was determined at 20 h p.i. Although all agents rapidly cleared from the blood, at 20 h p.i. blood levels and the levels in most organs of ¹²³I-fMLFK were significantly lower than those of the other three agents (P<0.05). The abscesses were clearly visualized with all agents from 4 h p.i. onwards. After 1 h p.i., the abscess uptake of ¹²³I-IL-1β was significantly higher than that of the other agents (P<0.05), with the highest uptake observed at 8 h p.i. (1.3%±0.3%). After 20 h p.i., the highest abscess-to-contralateral muscle ratios were obtained with ¹²³I-IL-1β, i.e. 39.0±11.5 vs 18.7±5.4, 18.1±2.3 and 29.9±7.0 for ¹²³I-IL-1α, ¹²³I-IL-1ra and ¹²³I-fMLFK, respectively. In conclusion, all agents localized in the infectious focus. The potential of radiolabelled IL-1β for imaging of infection was better than that of the other agents: higher absolute uptake in the infection and higher abscess-to-contralateral muscle ratios were obtained. The observation of localization of radiolabelled IL-1ra in infection was important since this protein can be administered to humans without any side-effects. Received 11 October and in revised form 27 December 1997     

F-labeling and biodistribution of the novel fluoro-quinolone antimicrobial agent, trovafloxacin (CP 99,219)

[¹⁸F]CP 99,219 [(1,5,6)-7-(6-amino-3-azabicyclo [3.1.0]hex-3-yl)-1-(2,4-difluorophenyl)-6-fluoro-1, 4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid] was prepared by ¹⁸F for ¹⁹F exchange followed by reverse-phase HPLC purification. Studies of the effects of reaction time and temperature on ¹⁸F incorporation demonstrated that heating 1.0 mg of CP 99,219 in 0.5 cc of DMSO with 4.5 mg of K₂CO₃ and 24 mg of Kryptofix for 15 min at 160 °C results in the optimal compromise between radiochemical yield and purity. This method routinely provides radiochemical yields of 15–30% [EOS] with radiochemical purities of >97%. Varying the concentration of CP 99,219 in the reaction mixture had no effect on yield. Biodistribution studies in rats demonstrated that significant concentrations of drug accumulate in most tissues. The tissues with the highest concentrations of drug were intestine, liver, kidney, and stomach.     

Subtyping of Y-chromosomal haplogroup E-M78 (E1b1b1a) by SNP assay and its forensic application

The continual discovery of new single-nucleotide polymorphisms (SNPs) has led to an increased resolution of the Y chromosome phylogeny. Some of these Y-SNPs have shown to be restricted to small geographical regions and therefore may prove useful in the forensic field as tools for the prediction of population of origin of unknown casework samples. Here, we describe a system for the molecular dissection of haplogroup E-M78 (E1b1b1a), consisting of multiplex polymerase chain reaction and minisequencing of M78 and nine population-informative Y-SNPs (M148, M224, V12, V13, V19, V22, V27, V32, V65) in a single reaction. Sensitivity and admixture studies demonstrated that the SNP protocol allows robust genotyping from as little as 50 pg of male DNA, even in the presence of 500-fold amounts of female DNA. In order to evaluate the suitability of E1b1b1a, subhaplogrouping for population-of-origin prediction, the distribution of E-M78 and its derived variants was determined in an Italian population sample (n = 326). Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

美托洛尔治疗心衰时脑利钠肽的动态变化

目的观察慢性充血性心力衰竭(CHF)患者接受选择性β受体阻滞剂美托洛尔治疗后血浆脑利钠肽(BNP)水平的动态变化,探讨美托洛尔在心衰预后中的临床意义。方法65例CHF患者随机分为美托洛尔组(34例)和对照组(31例)。两组均给予常规纠正心衰治疗(休息、限钠、洋地黄、利尿剂、ACEI和醛固酮拮抗剂),美托洛尔组在常规纠正心衰治疗基础上加服美托洛尔。观察两组治疗前及治疗5个月后临床症状、体征、心输出量(CO)、心脏指数(CI)、左室舒张末期内径(LVEDD)及左室射血分数(EF)等的改变情况,并采用免疫放射分析(IRMA)测定血浆BNP水平。结果治疗5个月后,两组症状、体征、CO、CI、LVEDD、EF均明显改善,血浆中BNP水平均较治疗前下降,且美托洛尔组血浆BNP水平(200.6±12.1ng/ml)明显低于对照组(120.7±14.1ng/ml,P<0.01)。结论CHF患者接受选择性β受体阻滞剂治疗后,可显著改善心脏内分泌功能,可能有助于改善心衰预后。     

低氧应激肽氢谱特征及急进高原的含量变化分析

目的分析血清低氧应激肽的特征和健康成年人急进高原前后血清低氧应激肽的改变,探讨其在高原病治疗和预后的作用。方法采用质谱和核磁波谱方法鉴定高原病患者体内存在特征性的应激肽,采集40例高原世居健康者为对照组,以40例健康成年人急进高原前、急进高原后24、72h及1周后血清,用化学显色试剂对应激肽含量进行测定。结果40例世居对照组应激肽含量为(729±98)mg/L;实验组健康成年人急进高原前、急进高原后24、72h和1周后应激肽含量分别为(735±117)mg/L、(908±103)mg/L、(917±101)mg/L和(828±95)mg/L。健康成年人急进高原前与急进高原后24h及72h之间有显著性差异(P<005),急进高原后24h人群与健康世居组之间有显著性差异(P<005),急进高原后1周与急进高原后24h之间有显著性差异(P<005)。结论应激肽作为一种特殊的应激蛋白,在急进高原前后有较大的改变,具有一定的检测价值。