运用组织芯片技术检测COX-2在结直肠癌中的表达及其临床意义

目的 探讨结直肠癌组织中COX-2的表达及其与各临床病理因素的关系，评价COX-2在结直肠癌预后判断中的价值。方法 应用组织芯片技术结合免疫组织化学SABC法，检测126例早中期结直肠癌组织中COX-2的表达情况，回顾性分析COX-2与各临床病理因素及预后之间的关系。结果 根据免疫组化染色强度，所有病例被分为COX-2高表达组和低表达组，其中高表达组有32例（25．4％），低表达组有94例（74．6％）。COX-2在结直肠癌中表达情况与年龄、性别、肿瘤大小、肿瘤部位、组织学类型、浸润深度、淋巴是否转移、Dukes分期均无相关。然而，高表达的COX-2与肿瘤复发、特别是血行转移显著相关（P〈0．05）。两组之间生存率具有显著差异（P=0．0067），COX-2高表达组术后五年生存率显著低于低表达组。多因素回归模型分析结果显示．在潜在的预后因素中（年龄、性别、肿瘤大小、肿瘤部位、组织学类型、淋巴是否转移、Dukes分期、COX-2表达），COX-2表达和Dukes分期可作为是结直肠癌根治术后独立预后因素，COX-2的检测可作为结直肠癌患者预后判断的一个有价值的指标。结论 高表达的COX-2与肿瘤复发、特别是血行转移显著相关，COX-2的检测町作为结直肠癌患者琐后判断的一个有价值的指标：应用组织芯片高效检测临床组织样本具有快速、方便、经济、准确的优点。     

SEN病毒D亚型ORF1-C端基因的克隆、表达和鉴定

目的:获得SENV-D亚型ORF1-C端蛋白基因序列,并进行表达,为进一步研究及诊断、治疗应用奠定基础.方法:用PCR法从SENV阳性血清中获得SENV-DORF1C端基因,测序验证后,构建了pQE30-SENV-D重组表达质粒,并经过转化E.coli M15,IPTG诱导表达,Western blot分析表达结果.结果:获得了正确序列的SENV-D亚型ORF1-C端蛋白基因序列,表达出Mr约为38×103的蛋白.表达蛋白能与患者血清中相应的抗体结合,发生抗原抗体反应.结论:成功获得并表达了SENV-D-ORF1-C端蛋白,并经Western blot印迹法证实能与阳性血清中的抗体发生抗原抗体反应,有可能用于检测SENV-D抗体.为今后进一步研究有助于疫情监测及早期发现感染者的抗体奠定了坚实的基础.     

Neurophysiological investigation of effects of the D-1 agonist SKF 38393 on tonic activity of substantia nigra dopamine neurons

The effects of the D-1 agonist SKF 38393 on tonic activity of rat substantia nigra pars compacta dopamine neurons were studied using extracellular, single-unit recording techniques. Unlike nonselective D-1/D-2 dopamine agonists or the D-2 agonist quinpirole, SKF 38393 did not inhibit dopamine neuronal activity when applied iontophoretically or when administered intravenously in doses up to 20 mg/kg to chloral hydrate-anesthetized rats. Moreover, pretreatment with SKF 38393 did not alter the inhibitory response of these neurons to apomorphine or the D-2 agonist quinpirole. However, in locally anesthetized, gallamine-treated, artificially respired rats, dopamine cell activity was significantly altered by i.v. administration of SKF 38393; firing rate increases and decreases were observed. Administration of the inactive enantiomer of SKF 38393, S-SKF 38393, did not induce similar changes in parallel experiments. These results support the idea that unlike D-2 autoreceptor stimulation, D-1 receptor stimulation does not exert a direct local effect on dopamine neurons in the substantia nigra pars compacta and suggest that D-1 receptor stimulation at sites postsynaptic to the dopamine cells may indirectly affect the activity of some dopamine neurons through long-loop feedback mechanisms.     

新抗喘药物的研究——茶碱衍生物及卤代苯乙醇胺化合物的合成

本文根据β_2-受体兴奋剂和茶碱类药物的作用机制,结合两类药物的基本结构和构效关系,运用新药设计中的拼合原理及生物电子等排原理,设计扦合成了两类共十六个尚未见于文献报导的化合物。经初步药理实验证明均具有抗哮喘作用,其中I_(5-8)活性较高,且对心脏副作用轻微,可望成为治疗哮喘的新药。     

新生儿脑病S-100蛋白监测与预后的研究进展

目的:通过监测新生儿缺氧缺血性脑病(HIE)患儿血清中S-100蛋白含量的动态变化,建立对HIE患儿的早期科学诊断及预后评估技术指标。并探讨S-100蛋白浓度变化与病情的严重程度及转归之间的关系。方法:采用酶联免疫吸附试验、双抗体夹心法检测,对120例HIE有症状组患儿(HIE轻度症状组40例,HIE中度症状组40例,HIE重度症状组40例)和40例新生儿窒息组患儿与40例对照组健康新生儿血清S-100蛋白含量进行检测,并进行动态观察。结果:①HIE有症状组不同时期血清S-100蛋白含量均显著高于对照组(P<0.01);②新生儿窒息组不同时期的血清S-100蛋白含量也明显高于对照组(P<0.01);③HIE有症状组血清S-100蛋白含量明显高于新生儿窒息组(P<0.05)。结论:HIE有症状组患儿病情越重,其血清S-100蛋白含量则越高,提示血清S-100蛋白可作为判断HIE病情程度的客观依据和早期诊断以及预后评估HIE的主要指标。     

Kringle 5的真核植物细胞穿梭表达载体的构建

目的：构建含有目的基因kring5的真核植物细胞穿酸表达载体。方法：应用RT－PCR方法，从正常人肝脏组织中获得kringle5基因，测序后，再通过DNA重组技术，将kringle5基因片段克隆到真核植物细胞表达载体pBI121和pCAMBIA3301上。结果：得到的kringle5基因序列测定结果与文献相符，重组载体pB1k5和pC33k5经酶切鉴定正确，含有植物高效表达CaMV35S启动子。结论：重组载体pB1k5和pC33k5不仅可以在大肠杆菌中稳定复制，而且可以在植物细胞中表达。     

硬膜外腔阻滞对胸部手术应激反应的影响

目的 观察硬膜外腔阻滞对胸部手术应激激素和细胞因子的影响。方法20例食管癌手术病人,随机分为两组,每组10例,即全麻复令硬膜外腔阻滞(GEA)组和全麻(GA)组,分别测定麻醉诱导前、手术2h、手术4h、术毕、术后1d及术后3d的血浆去甲肾上腺素、肾上腺素、血清促肾上腺皮质激素(ACTH)、皮质醇、C-反应蛋白、IL-6及IL-10的水平。结果 血浆去甲肾上腺素和血清皮质醇GEA组术中术后无显著改变,但GA组术毕和术后1d显著升高(P<0.05),术后3d恢复至术前水平,组间比较前者有显著差异(P<0.05)。两组血浆肾上腺素、IL-10术中术后均无显著变化。两组血清ACTH、IL-6及CRP术中术后均显著升高(P<0.05),组间比较无显著差异。结论 硬膜外腔阻滞可以减轻胸部手术的应激反应。IL-6是较CRP更灵敏的反映组织损伤的炎性指标。     

Species- and sex-specific variations in binding of ochratoxin A by renal proteins in vitro

The mycotoxin ochratoxin A (OTA) is a potent renal carcinogen in rodents and induces renal fibrosis in pigs. Furthermore, OTA has been associated with the development of renal tumors and nephropathies in humans. Large species- and sex-differences are observed in sensitivity toward OTA-mediated toxicity and carcinogenicity, yet neither the mechanism(s) resulting in OTA toxicity nor the reasons for the observed species- and sex-specificities are known. This paper investigated variations in OTA handling viz binding to renal proteins which could possibly explain the observed differences in OTA susceptibility in vivo and in vitro. The results obtained via a modification of a standard receptor-binding assay demonstrated the presence of at least one homogeneous binding component in renal cortical homogenates from pig, mouse, rat and humans. This component was shown to bind OTA in a specific and saturable manner. A range of compounds selected for their affinity for steroid receptors and/or for various known organic anion transporters were employed in a competition assay to answer the question whether this homogenous OTA binding component represents a steroid-like receptor component or one of the known organic anion transporters of the kidney. Although many of the compounds were able to compete with OTA for protein-binding, the competition patterns displayed a distinct species specificity and did not correspond to the competition patterns associated with presently known organic anion transporters of the kidney in the mouse, rat or human. The data thus suggests the presence of a new organic anion transporter or more likely, a cytosolic binding component of unknown function with high affinity and capacity for OTA binding in humans, rats, mice and possibly pigs.     

High-protein Weight-loss Diets: Are They Safe and Do They Work? A Review of the Experimental and Epidemiologic Data

Recommendations for increased consumption of protein are among the most common approaches of popular or fad diets. This review summarizes the effects of dietary protein on satiety, energy intake, thermogenesis, and weight loss, as well as its effect on a variety of health outcomes in adults. In short-term studies, dietary protein modulates energy intake via the sensation of satiety and increases total energy expenditure by increasing the thermic effect of feeding. Whereas these effects did not contribute to weight and fat loss in those studies in which energy intake was fixed, one ad libitum study does suggest that a high-protein diet results in a greater decrease in energy intake, and therefore greater weight and fat loss. In terms of safety, there is little long-term information on the health effects of high-protein diets. From the available data, however, it is evident that the consumption of protein greater than two to three times the U.S. Recommended Daily Allowance contributes to urinary calcium loss and may, in the long term, predispose to bone loss. Caution with these diets is recommended in those individuals who may be predisposed to nephrolithiasis or kidney disease, and particularly in those with diabetes mellitus.