复合微量营养素对C57BL糖尿病小鼠Th1、Th2型细胞因子表达的调控作用

目的通过研究复合微量营养素(MC)对C57BL糖尿病小鼠Th1、Th2型细胞因子抗原表达的影响,探讨AM拮抗糖尿病的分子作用机制。方法应用小剂量多次注射STZ的方法(multiplelowdoseofstreptozotocin,MLDS)制备小鼠IDDM模型,经灌胃联合投予硒(Se)、维生素E(VE)、钒(V)、铬(Cr),应用流式细胞术观察表达Th1型细胞因子(TNFα、IFNγ)和Th2型细胞因子(IL4、IL10)抗原的淋巴细胞数百分比。结果IDDM小鼠外周血表达TNFα、IFNγ抗原的淋巴细胞数百分比明显升高(P<001),而外周血、脾组织表达IL10抗原的淋巴细胞数百分比明显降低(P<001);应用MC可显著降低IDDM小鼠外周血淋巴细胞TNFα的表达(P<001),显著升高外周血淋巴细胞IL10、脾淋巴细胞IL4的表达(P<001,P<005)。结论MC可通过下调Th1型细胞因子、上调Th2型细胞因子的基因表达,来调节IDDM时的免疫失衡,减轻胰岛自身免疫性损害,拮抗糖尿病的发生发展。     

玻璃体切除联合眼内填充治疗复杂视网膜脱离

目的　探讨复杂性视网膜脱离应用玻璃体切除联合眼内填充手术的效果及并发症的发生情况。方法　对2 9例伴有眼球破裂伤、眼内炎、玻璃体积血、白内障、PVR或糖尿病性视网膜病变等的复杂性视网膜脱离进行玻璃体视网膜手术的连续病例临床资料进行回顾性分析。结果　玻璃体切除联合玻璃体腔注气术12眼,1次手术视网膜复位8眼(66.67%) ;玻璃体切除联合玻璃体腔内硅油填充术2 0眼(包括注气失败的3眼) ,1次手术完全复位13眼(65 .0 0 %)。统计学检验两组1次手术视网膜复位率的差异无显著意义。术后视力较术前提高者2 1眼(72 .41%)。常见的并发症有复发性视网膜脱离、并发性白内障及继发性青光眼等。结论　玻璃体切除联合眼内填充能有效地治疗复杂性视网膜脱离,PVR仍然是术后视网膜脱离复发的主要因素。     

复杂视网膜脱离视网膜下膜超微结构研究

目的研究复杂视网膜脱离病例中视网膜下膜的超微结构特征,探讨视网膜下膜的细胞成分。方法对21例复杂孔源性视网膜脱离伴视网膜下膜者行玻璃体切割术加视网膜下膜切除术,将所获得的视网膜下膜经处理后于光镜下选择细胞较密集处做超薄切片,染色后行透射电镜观察并摄影。结果复杂视网膜脱离视网膜下膜中,色素上皮细胞多呈散在分布,未见明显基底膜供细胞附着,在条索或片状膜当中的色素上皮细胞形态存在变化。神经胶质细胞胞浆内有较丰富的细胞器和直径约10nm的中间型微丝形成。视网膜下膜的细胞间质含有大量胶原纤维。成纤维细胞形状不规则,活跃,胞浆中见大量直径4～6nm微丝。结论视网膜下膜主要由视网膜色素上皮细胞、神经胶质细胞、成纤维细胞和胶原纤维组成。视网膜下膜中色素上皮细胞、神经胶质细胞和成纤维细胞有转化与增殖能力。     

甘珀酸钠苦参碱包合作用的研究

 目的制备甘珀酸钠和苦参碱的包合物,研究甘珀酸钠和苦参碱的包合作用,测定了包合物的包合比。方法分别配制甘珀酸钠和苦参碱溶液,将两种溶液按不同比例混合,形成包合物。用X射线粉末衍射,红外光谱和核磁共振谱来表征包合物,用紫外可见分光光度法确定包合物的形成常数,用循环伏安法对包合物的电化学性质进行了研究。结果XRD,IR,¹H-NMR等分析数据及熔点、比旋光度的变化确证了包合物的形成。结论甘珀酸钠和苦参碱可以形成1∶1的包合物。     

枳实对大鼠小肠电活动影响的下丘脑作用机制研究

目的观察枳实对在体大鼠小肠电活动的影响,探讨其作用与下丘脑内脑肠肽胆囊收缩素(CCK)、生长抑素(SS)的相关性。方法采用消化道腔外电极记录法观察枳实组与生理盐水组移行性综合肌电(MMC)周期时程、活动期/周期比例、活动期每分钟快波数的变化；采用免疫组织化学法结合显微图像定量分析扫描系统检测下丘脑背内侧核(DMH)、腹内侧核(VMH)、下丘脑外侧区(LHA)CCK的相对含量,以及下丘脑VMH、LHA、室旁核(PVN)SS的相对含量。结果枳实组与生理盐水组比较MMC周期缩短、活动期/周期比例增大、活动期每分钟快波数增加,差异有显著性；与生理盐水组及空白对照组比较枳实组DMH、VMH、LHA的CCK阳性神经元表达降低；VMH、LHA、PVN的SS阳性神经元表达增高,与生理盐水组及空白对照组比较差异有显著性。结论枳实对大鼠小肠电活动有兴奋作用,这种兴奋作用可能与下丘脑内的CCK、SS有关。     

醋柳黄酮β-环糊精包合物的制备、鉴别与测定

目的:制备醋柳黄酮β-环糊精包合物,以增加醋柳黄酮的水溶性,改善其制剂制备性能。方法:用β-环糊精饱和水溶液法制备包合物,薄层色谱法对黄酮类有效成分进行鉴别并对形成的包合物进行验证,用紫外分光光度法测定原料药及包合物中药物的含量。结果:制备成淡黄色粉末状醋柳黄酮包合物,包合物的收得率为62.17%,平均包合率为9.49%。结论:醋柳黄酮中的黄酮类有效成分能被β-环糊精包合,具有较高的包合率。     

清肺化痰通络方对大鼠肺纤维化肺组织中iNOS表达的影响

目的：观察清肺化痰通络方对博莱霉素所致大鼠肺纤维化中诱导型一氧化氮合酶（iNOS）表达的影响。方法：通过气管内灌注博莱霉素A5（BLMA5）复制大鼠肺纤维化动物模型，于灌服该方后3、7、14、28天，分别观察肺组织病理变化及iNOS的表达。结果：清肺化痰通络方能显著改善大鼠的一般身体状况，减轻病变的程度，并抑制iNOS的表达。结论：清肺化痰通络方可能通过抑制iNOS的异常表达。对BLMA5致肺组织纤维化起到一定的防治作用。     

Quinolinic acid enhances permeability of rat brain microvessels to plasma albumin

Several studies have established that increased cerebrospinal fluid (CSF) levels of quinolinic acid (QUIN), a macrophage/microglia-derived excitotoxin with N-methyl-D-aspartate (NMDA)-receptor affinity, may reflect abnormal blood-brain barrier (BBB) function in patients with acquired immunodeficiency syndrome (AIDS) dementia complex, exhibiting a relationship to their clinical and neurological status. This study was aimed to evaluate whether QUIN (250 nmol/0.25 microl/ventricle) infused into both lateral cerebral ventricles permeates adult rat brain microvessels to plasma albumin. Possible BBB dysfunction was examined 4 days after the intracerebroventricular (i.c.v.) infusion of QUIN by measuring plasma albumin extravasation using rocket immunoelectrophoresis. The i.c.v. infusion of QUIN failed to increase the extracellular tissue concentration of albumin in the entorhinal cortex, but significantly higher levels were found in the hippocampus proper (but not in the subiculum region and dentate gyrus) and in the striatum. To evaluate the possible relationship between plasma protein extravasation and QUIN-induced tissue necrosis, we quantified neuronal death in the rat hippocampal formation (subiculum, CA1/CA3 areas of the hippocampus proper, dentate gyrus). We found significantly higher tissue levels of plasma albumin in the hippocampus proper, in which the CA1 area exhibited the highest neuronal loss while the low rate of neuronal death was not accompanied by significant albumin extravasation in the dentate gyrus. However, in case of the subiculum, in which the neuronal loss reached comparable values to those in the CA1 area, we did not find significant enhancement of plasma albumin leakage into this area. The regional differences in brain microvascular permeability may depend on the density of NMDA receptors in the multicellular capillary barrier, but the differences in neuronal death may also reflect an involvement of NMDA receptors in neuronal membranes. We conclude that increased CSF QUIN levels evoke a dysfunction of the BBB that may only partially be related to sites with pronounced neuronal damage in the rat brain regions susceptible to NMDA-receptor mediated toxicity.     

麦考酚酸高产菌株的选育

以短密青霉MA-4为出发菌株，应用紫外和亚硝基胍诱变，结合麦考酚酸与三氟乙酸抗性筛选，获得高产、稳定的突变株MA—H8。经过发酵培养基优化，突变株发酵水平为4500μg／ml。     

Improved detection of melanoma antigen-specific T cells expressing low or high levels of CD8 by HLA-A2 tetramers presenting a Melan-A/Mart-1 peptide analogue.

MHC class I tetramers containing peptide epitopes are sensitive tools for detecting antigen-specific CD8(+) T-cell responses. We demonstrate here that binding of HLA-A2 tetramers to CD8(+) T cells specific for the melanoma-associated antigen Melan-A/MART-1 can be fine-tuned by altering either the bound peptide epitope or residues in the alpha 3 domain of HLA-A2, which is important for CD8 binding. Antigen-specific T cells expressing high levels of CD8 could be detected using HLA-A2 tetramers containing the peptide AAGIGILTV, an epitope which is naturally processed and presented from Melan-A/MART-1. In contrast, low CD8-expressing, antigen-specific T cells could be detected efficiently only by using a mutated HLA-A2 tetramer with an altered CD8 binding site or, less efficiently, using the wild-type HLA-A2 tetramer loaded with the peptide analogue ELAGIGILTV, which is superior in stimulating antigen-specific T-cell responses. Our results suggest ways to optimize the identification and expansion of antigen-specific T cells with different requirements for the costimulatory CD8 molecule in facilitating T-cell receptor binding to peptide variants.