Poly-L-Lysine玻片在寡核苷酸芯片制备中的改进

目的 为了制得适合固定未修饰寡核苷酸的芯片，提高检测灵敏性，对Patrick Brown实验室的多聚左旋赖氨酸包被玻片的方法进行改进。方法 玻片经清洗后用缩水甘油-丙氧基三甲氧基硅烷进行硅烷化，然后应用Poly-L-Lysine在玻片表面形成聚合物涂层，经次亚苯基二异硫氰酸盐表面活化后可使寡核苷酸共价连接在芯片表面。设计了各种实验考察方法改进前后芯片表面的性能，并将改进后的玻片初步应用于SARS冠状病毒寡核苷酸芯片检测中。结果 方法改进后芯片表面性能优良：固定效率高、点的同一性好、杂交效率和热稳定性好、寡核苷酸结合牢固、芯片可以重复利用。结论 利用共价连接，方法改进后的芯片表面适合固定未修饰的寡核苷酸，解决了寡核苷酸与玻片之间物理结合不稳定、易剥离的缺陷，提高了芯片检测的灵敏性。     

Reduced expression of CD69 and adhesion molecules of T lymphocytes in asthmatic children receiving immunotherapy

T lymphocytes play a fundamental role in the initiation and regulation of chronic inflammatory responses in patients with asthma. CD69 is an early marker of T‐cell activation. The levels of intercellular adhesion molecule‐1 (ICAM‐1, CD54) and L ‐selectin have been reported to increase in patients with allergic diseases and asthma. The present study was therefore undertaken to investigate the expression of CD69, CD54, and L ‐selectin by T lymphocytes of children with asthma, before and after immunotherapy. Eighteen children newly diagnosed with asthma, 11 good and nine poor responders to immunotherapy, and 16 normal subjects, were enrolled in this study. The percentages of CD69⁺, CD54⁺, and CD62L⁺ cells in T lymphocytes were measured by using flow cytometry. The levels of CD69, CD54, and CD62L in serum and culture supernatants were determined by using enzyme‐linked immunosorbent assay (ELISA). The expression of CD69 and CD54 on CD3⁺ T lymphocytes was significantly higher in children with asthma than in control patients. All the patient groups expressed (spontaneously and following stimulation with phorbol myristate acetate and ionomycin together with mite‐extract proteins) greater amounts of CD69 and CD54 than did control subjects. With long‐term immunotherapy, the percentages of CD69⁺ and CD54⁺ T lymphocytes were significantly lower in patients with a good response to immunotherapy. Our results also showed significantly lower serum L ‐selectin levels following immunotherapy. In conclusion, successful immunotherapy resulted in decreased expression and production of CD69 and CD54. These results may explain, in part, the clinical efficacy of immunotherapy.     

固相板酶免法测定人血清游离三碘甲状腺原氨酸

目的 :探讨游离三碘甲状腺原氨酸 (FT3)的酶免疫测定法对于诊断甲状腺功能亢进和监测甲状腺激素补充治疗效果及预后情况的重要意义。方法 :采用聚苯乙烯96孔板作为固相载体 ,利用亲和纯化的兔抗T3 抗体和T3 - β-半乳糖苷酶标记物 ,建立了固相板竞争法。结果 :可测范围为0.48～45.0ng/L ,灵敏度0.13ng/L ,平均批内变异系数为 (n=20)6.4 % ,平均批间变异系数为 (n=8)12.4% ,正常参考范围4.14±0.97ng/L。正常人与甲亢病人能明显分开。结论 :酶免法测定血清FT3,其灵敏度、精密度均达到放免水平 ,且可避免同位素污染     

失眠症患者心理防御机制与社会支持的相关研究

目的　探讨失眠症患者心理防御特征和社会支持 ,并分析两者的关系。方法　采用心理防御方式问卷 (DSQ )、社会支持评定量表 (SSRS)对失眠症患者和正常健康者各 5 0例进行评定。结果　患者组心理防御方式 (不成熟型、中间型防御因子和掩饰因子的因子分高 ,成熟防御因子分低 )和社会支持 (社会支持总分、主观支持分、对支持利用度低 )与对照组相比差异有显著性 (P <0 .0 5或 0 .0 1) ;相关分析发现 ,患者组不成熟防御机制与SSRS的支持总分、主观支持分、对支持利用度呈负相关 (P均 <0 .0 5 ) ,中间型防御机制与SSRS的主观支持呈负相关 (P <0 .0 5 )。结论　失眠症患者心理防御机制的成熟程度较正常人低 ,社会支持差 ,两者存在一定的关系。     

消化道肿瘤血浆胃动素测定及其意义

作者报告经病理诊断消化道恶性肿瘤患者30例及手术切除肿瘤患者15例的血浆胃动素水平,并以健康成人31例作为对照。消化道恶性肿瘤的胃动素水平373.4±123.7ng/L,显著高于正常对照组(162.3±52.3ng/L,P<0.001),而消化道恶性肿瘤患者术前及术后胃动素水平无显著差异。消化道恶性肿瘤患者胃动素水平增高的原因可能是肿瘤刺激神经所致。胃癌和结肠癌细胞可产生胃泌素样物质,它可刺激胃动素的释放。     

Compensatory adaptation of the heart to chronic rate overload: increase in calcium transport ATPase activity of myocardial sarcoplasmic reticulum

This study was undertaken to test the hypothesis that a compensatory response of the heart to a chronic and continuous, metabolic and heart rate overload was an increase in the calcium sequestering activity of the myocardial sarcoplasmic reticulum. Calcium sequestering activity was estimated by determination of the calcium-dependent ATPase (Ca2+-ATPase) activity of isolated microsomes. Chronic rate overload was modelled by comparing: dysthyroid and control rats; control swine and swine with implanted cardiac pacemakers set at 180 beats/min; and different species of mammals with widely different heart rates. The myocardial sarcoplasmic reticulum Ca2+-ATPase pump activity was significantly increased by 39% for hyperthyroid rats compared to control rats and by 87% for control rats compared to thyroidectomized rats; by 63% for paced swine compared to control swine; and by 43% for rats compared to guinea pigs, by 140% for guinea pigs compared to dogs and by 120% for dogs compared to cows. These data indicate that calcium sequestering activity of myocardial sarcoplasmic reticulum increases in equivalent proportion to the chronotropic demand and that heart rate is a hemodynamic correlate of the sarcoplasmic reticulum Ca2+-ATPase activity.     

病理性损伤因素对肾小管上皮细胞表型转化的影响

目的 :观察低血清、高糖、高蛋白刺激下肾小管上皮细胞向间叶性细胞的表型转化 ,探讨损伤的肾小管上皮细胞在肾小管间质纤维化中的作用。方法 :以体外培养的人近端肾小管上皮细胞为对象 ,分别用含低血清(0 .2mol·L-1小牛血清 )、高葡萄糖 (4.5 g·L-1)、高白蛋白 (15 g·L-1)的培养液培养 7d ,透射电镜检测细胞形态变化 ;免疫组化和Westernblot检测细胞表型改变 ,包括CK、Vimentin、α SMA、Ⅰ和Ⅲ型胶原 ,原位杂交检测Ⅰ型胶原mRNA表达 ,同时检测细胞TGFβ1蛋白表达。 结果 :低血清、高糖、高蛋白直接刺激下 ,肾小管上皮细胞出现明显形态学改变 ,包括细胞变为长梭形 ;透射电镜下细胞内线粒体明显减少 ,粗面内质网明显增加 ,并出现actin样微丝。免疫组化染色和Westernblot显示CK表达明显减弱 ,Vimentin、Ⅰ、Ⅲ型胶原、α SMA和TGFβ1表达增强。原位杂交显示Ⅰ型胶原mRNA表达增强。结论 :低血清、高糖、高蛋白可直接引起人近端肾小管上皮细胞TGFβ1表达增加 ,并发生向间叶性细胞的表型转化     

自体骨髓多次注射治疗家兔骨缺损的实验研究

目的：观察骨髓多次注射的成骨性能。方法：利用家兔实验性骨缺损模型，通过大体、组织学观察，生物力学测定，评价骨髓不同移植次数促进骨缺损愈合的情况。结果：骨髓单次移植能够促进骨缺损的愈合，然而，骨髓二次移植组骨折愈合程度明显优于骨髓单次移植组，但骨髓三次移植组与骨髓二次移植组骨折愈合无明显差异。结论：在骨折愈合早期增加骨髓移植次数，能明显增强骨髓的成骨能力，加快骨折愈合     

授权影印西文书的特点及其机读目录的著录方法

对授权影印书的特点进行了分析，并对其机读目录著录方法和需要注意的问题进行了论述。     

逆行灌注心脏不停跳心肌保护的实验研究

目的：探讨逆行灌注心脏不停跳心肌保护的效果。方法：中国大白兔32只随机分为4组，每组8只，A组（对照组），B组（心脏不停跳组，血温34℃），C组（温血心脏麻痹液组，血温34℃），D组（冷血心脏麻痹液组，血温4℃）。采用离体兔心左室做功模型，经冠状静脉窦逆行灌注稀释血液或血停搏液60min。比较观察心功能、冠脉排出液心肌酶谱、心肌组织乳酸、心肌细胞内游离钙浓度、心肌细胞膜ATP酶活性、心肌细胞膜脂质过氧化产物丙二醛含量的变化。结果：B组心功能恢复、心肌细胞膜ATP酶活性显著优于C组和D组，C组优于D组；B组心肌酶漏出、乳酸生成量、心肌细胞内游离钙浓度、心肌细胞膜丙二醛（MDA）含量低于C组和D组，C组低于D组。结论：常温氧合血逆灌心脏不停跳良好的心肌保护效果与其减轻自由基损伤，较好地保持细胞内外Ca^2 平衡，减轻细胞内钙超载，保护细胞膜酶功能有关。