抗醛糖还原酶单克隆抗体的制备与特性鉴定

目的:制备抗醛糖还原酶(AR)的单克隆抗体(mAb),并与本室制备的抗醛糖还原酶相似蛋白(ARL-1)mAb进行比较。方法:经RT-PCR获得AR基因,将基因插入pGEX-4T-1(His)6C中,构建重组质粒pGEX-4T-1(His)6C-AR,以重组质粒转化E.coliRosetta诱导表达GST-AR蛋白。以纯化的GST-AR蛋白免疫BALB/c小鼠,采用杂交瘤技术制备mAb。应用间接ELISA和Western blot方法对mAb进行筛选和鉴定。使用Clustalx和Antheprot软件,比较AR与ARL-1的同源性,表达GST-dAR[80~142氨基酸(aa)],与ARL-1差异较大;并分析AR的抗原性,表达GST-dA1(1~79aa)、GST-dA2(80~99aa)、GST-dA3(111~142aa)、GST-dA4(143~316aa)。利用AR全长及截短蛋白,采用Western blot分析制备的抗AR mAb识别AR抗原的部位。结果:获得3株稳定分泌抗AR mAb的杂交瘤细胞系ARB3、AR7B3G4和ARF10。3株抗GST-AR的mAb均为IgG1(κ型),腹水mAb效价为1∶4×105,细胞培养上清mAb效价为1∶1×104,3株mAb均可与胎盘组织中的AR蛋白起反应,而与GST-ARL-1和GST蛋白无交叉反应。它们分别为抗GST-dA1、GST-dA3和GST-dA4蛋白的mAb。结论:成功地制备了3株特异性抗AR mAb,可分别识别AR的1~79、111~142、143~316位氨基酸。将它们与抗ARL-1mAb联合应用,将有助于进一步研究AR与ARL-1蛋白的功能,并为深入探讨AR、ARL-1与相关疾病的关系及进行大规模的流行病学调查提供了有力的工具。     

膜表达热休克蛋白70真核表达载体的构建及其对肿瘤细胞的转染

目的构建普遍适合性的膜表达热休克蛋白70(HSP70)的真核表达载体,修饰并建立HSP 70表达并结合在细胞膜表面的肿瘤细胞.方法RT-PCR法从人胚肾中获取HSP 70的cDNA,扩增成带酶切位点的目的基因.选择载体pDisplay,将目的基因插入载体多克隆酶位点中.DNA测序证实后,以脂质体转染的方法,将载体转染到黑色素瘤细胞.用G418抗性筛选获得阳性克隆.结果酶切电泳和DNA测序证实载体构建正确;目的基因的上游带信号肽序列,下游带跨膜序列.RT-PCR产物电泳、Westem Blotting、激光共聚焦显微镜和流式细胞术证实大量HSP 70表达并结合在转染的黑色素瘤细胞膜表面.结论成功地构建了膜表达HSP 70的真核表达载体;转染到黑色素瘤细胞后,细胞膜表面表达丰富的HSP70.转染细胞可以进一步制备新型瘤苗.     

Bleeding from foveolar hyperplasia developing on a gastrointestinal stromal tumor of the stomach

A 52‐year‐old Japanese woman who presented with gastrointestinal (GI) bleeding underwent a proximal gastrectomy for a gastrointestinal stromal tumor (GIST) with a foveolar hyperplasia at the apex of the tumor, 4.5 cm in size, located in the upper body of the stomach. Although GIST are often asymptomatic and are found only incidentally, clinical symptoms such as bleeding, abdominal pain, or obstruction, occasionally lead to a premorbid diagnosis. When submucosal tumors present GI bleeding, the source of the bleeding usually is an ulceration of the mucosa over the tumor. However, in the present study, it was thought that the bleeding originated from the region of foveolar hyperplasia.     

妊高征患者胰岛素样生长因子水平及对胎儿生长发育的影响

目的：探讨妊娠高血压综合征(妊高征)患者血清胰岛素样生长因子(IGF-1)和胰岛素样生长因子结合蛋白-3(IGFBP-3)水平与疾病程度及新生儿出生体重之间的关系。方法：用放射免疫方法测定33例妊高征和32例正常血压妊娠妇女的血清IGF-1、IGFBP-3的水平。结果：重度妊高征组IGF-1显著低于正常组，IGFBP-3在各组间的水平浓度差异均无显著性，但IGF-1与IGFBP-3之间呈正相关。新生儿出生体重随妊高征严重程度的加剧而降低，各组间比较差异有显著性(F=5．453，P=0．002)。结论：妊高征患者的发病及严重程度与IGF-1有明显的关系，IGF-1与胎儿的发育及新生儿出生体重有明显的相关性。     

高压氧对冠心病患者C-反应蛋白的影响

目的　了解高压氧(HBO)治疗对冠心病患者C 反应蛋白 (CRP)的影响。方法　冠心病患者共 60例,随机分为HBO治疗组(HBO组)30例和对照组 30例。HBO治疗压力 0. 2MPa,每日 1次,每周 6次,共治疗 4周。比较治疗前后两组CRP的测定结果。结果　HBO组治疗后CRP与治疗前差异有显著性意义(P<0. 001);治疗后两组CRP差异有显著性意义(P<0. 001);治疗前两组CRP差异无显著性意义(P>0. 05);对照组治疗前后CRP差异无显著性意义 (P>0. 05)。结论　HBO治疗可显著降低冠心病患者的CRP含量。     

急性冠脉综合征患者循环内皮祖细胞与炎性相关因子的研究

目的:探讨急性心肌梗死(AMI)、不稳定性心绞痛(UA)患者外周血中循环内皮祖细胞(EPCs)与炎性相关因子C反应蛋白质(CRP)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的相关性。方法:取AMI(25例)UA患者(28例)及健康体检者(对照组32例)静脉血,采用密度梯度离心法从外周血获得单个核细胞;激光共聚焦显微镜鉴定FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPCs,并在倒置荧光显微镜下计数;酶联免疫法检测各种炎性相关因子。结果:AMI组及UA组的EPCs数量和血清CRPI、L-6、TNF-α水平显著高于正常对照组。结论:AMI及UA患者EPCs数量增加与炎性相关因子增加有关。     

Post–traumatic Alzheimer''s disease: preponderance of a single plaque type

The cause of Alzheimer's disease is unknown. Several factors have been proposed including head trauma. At present, the link between head injury and a subsequent neurodegenerative process is largely circumstantial, except in the case of dementia pugilistica (punch drunk syndrome) found in boxers. Recent studies have shown that the brains of boxers with this syndrome contain large numbers of 'diffuse' beta-protein immunoreactive plaques. We supposed that this plaque type might be associated with trauma induced Alzheimer-like degeneration. In order to test this hypothesis we have re-investigated a previously reported case of post-traumatic premature Alzheimer's disease. Immunocytochemistry using antibodies to amyloid beta-protein revealed large numbers of 'diffuse' non-Congophilic plaques with little or no neuritic component. A similar preponderance of this plaque type is present in the brains of boxers with dementia pugilistica. Our observations support the idea of a trauma induced Alzheimer-like degenerative process and indicate that such a condition is associated with a marked preponderance of 'diffuse' plaques.     

Identification of a Membrane Receptor for Retinol-Binding Protein Functioning in the Cellular Uptake of Retinal

Retinol-binding protein (RBP) is the transport protein that carries retinol in the circulation from the liver to its target tissues. The existence of a cell-surface receptor on the target cells, which mediates the uptake of retinol from RBP, has been known since 1975. Recently, it was identified as an integral transmem-brane protein named STRA6 that is inducible by retinoic acid in certain cancer cells. The receptor was found to be highly specific for RBP, with high affinity, and to be localized in all tissues known to require retinol for their function, particularly the pigment epithelium of the eye.     

Stereoselective plasma protein binding of ibuprofen enantiomers

Summary We have developed a novel and reproducible method for determining the plasma protein binding of the two ibuprofen enantiomers in the presence of each other. The method involves the use of radiolabelled racemic ibuprofen, equilibrium dialysis, derivatization of the enantiomers to diastereomeric amides, high-performance liquid chromatography, and radiochemical analysis.We have determined the plasma protein binding of R(–)- and S(+)-ibuprofen in 6 healthy male volunteers after the oral administration of 800 mg racemic ibuprofen.The mean time-averaged percentage unbound of the R(–)-enantiomer, 0.419 was significantly less than that of the S(+)-enantiomer, 0.643, consistent with stereoselective plasma protein binding.The percentage unbound of each ibuprofen enantiomer was concentration-dependent over the therapeutic concentration range and was influenced by the presence of its optical antipode.