Insulin-like growth factor (IGF)-I,-II and IGF-binding proteins in the cyst fluid of a patient with astrocytoma

Insulin-like growth factor (IGF)-I,-II and IGF-binding proteins (IGFBPs) were demonstrated in the cyst fluid of a patient with a hypothalamic astrocytoma. The astrocytoma cyst fluid was subjected to gel chromatography at low pH and the IGF-I and IGF-II levels were measured by specific radioimmunoassays. Immunoreactive IGF-I and IGF-II levels were 19 ng/ml and 78 ng/ml respectively. Several-fold higher IGF-II values were obtained when cyst fluid was not extracted or was extracted with acid ethanol before radioimmunoassay analysis. The immunoreactive IGFBP-1 concentration was 26 ng/ml. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and subsequent Western ligand blotting with [¹²⁵I]IGF-II revealed bands at 200, 34.5, 29.5, 24 and 21 kD as visualized by autoradiography. Binding studies demonstrated that these binding proteins bind specifically [¹²⁵I]IGF-I and [¹²⁵I]IGF-II. These observations suggest that IGFs as well as IGF-binding proteins are produced by astrocytoma cells and may act in a paracrine or autocrine fashion capable of modulating the growth of astrocytoma tumours.     

金黄色葡萄球菌NorA蛋白免疫检测方法的建立

目的建立金黄色葡萄球菌耐药转运蛋白NorA免疫检测方法。方法利用琼脂稀释法测定诺氟沙星（NFLX）对临床分离的6株金葡菌和标准菌株ATCC25923的MIC。对norA基因进行克隆、表达，提取表达产物包涵体，并进行了纯化。将纯化的表达蛋白作为免疫原免疫家兔，获取NorA蛋白的抗体血清。利用制得的抗体通过Western blotting检测临床分离的6株金葡菌的NorA蛋白表达水平。结果氟喹诺酮类药物耐药水平较高的临床分离金葡菌菌株的NorA蛋白表达水平一般较高，敏感菌株的耐药水平较低且接近。结论免疫检测方法可用于检测金葡萄菌耐药转运蛋白NorA的表达水平。     

1426例幽门螺杆菌抗体检测的临床价值探讨

目的:探讨I型和II型幽门螺杆菌(Helicobacter Pylori,简称 HP)与小儿腹痛的关系.方法:对1426例腹痛患儿,用免疫印迹法进行分型检测.结果:检出HP阳性590例,占41.37%;其中I型HP感染342例,占57.97%;II型248例,占42.03%.结论:免疫印迹法分型检测HP对临床诊治具有重要的指导意义.     

静脉麻醉药对脂多糖诱导的神经星形胶质细胞增殖的影响及其机制

目的 检测静脉麻醉药对脂多糖诱导的神经星形胶质细胞的影响及其机制。 方法 利用MTT实验、集落形成实验和Annexin V-FITC/PI双染色法检测静脉麻醉药对脂多糖作用后的神经星形胶质细胞增殖和凋亡的影响。Western blotting技术检测相关机制。 结果 静脉麻醉药可以逆转脂多糖诱导的神经星形胶质细胞增殖。经麻醉药处理后的神经星形胶质细胞p-Akt和p-mTOR蛋白表达下降，Caspase-3和Caspase-9表达水平升高。结论 静脉麻醉药可以通过抑制mTOR通路逆转脂多糖诱导的神经星形胶质细胞增殖。     

Presence of hBD-1 and hBD-2 in human cerumen and external auditory canal skin

Conclusion. Human β-defensin-1 (hBD-1) and human β-defensin-2 (hBD-2) antimicrobial peptides present in the cerumen, which is composed of exfoliated epithelial keratin and gland secretion, might provide the first line of defense against microbes in external auditory canal (EAC) skin. Objectives. Cerumen (earwax) plays a primary role in protecting the EAC skin and tympanic membrane. Even though the protection by antimicrobial peptides present in the skin secretion has been well established, little is known about the intrinsic role of the peptides in the EAC skin and cerumen. The aim of this study was to examine the presence of important antimicrobial peptides, hBD-1 and hBD-2, in the cerumen and EAC skin. Materials and methods. Cerumen was collected from 20 healthy adults, and the EAC skins were obtained from 12 patients who underwent middle ear surgery with canaloplasty. The presence of hBD-1 and hBD-2 was analyzed using immunohistochemistry and Western blotting. Results. In the immunohistochemical study of the EAC skin, expression of hBD-1 and hBD-2 was observed in both the epithelium and the glands. The presence of hBD-1 and hBD-2 peptides in the cerumen was confirmed by Western blotting.     

CCM2 expression during prenatal development and adult human neocortex

Cerebral cavernous malformation (CCM) is one of the most common types of vascular malformations of the central nervous system, affecting nearly one in 200 people. CCM lesions are characterized by grossly dilated vascular channels lined by a single layer of endothelium. Genetic linkage analyses have mapped three CCM loci to CCM1, CCM2 and CCM3. All three causative genes have now been identified allowing new insights into CCM pathophysiology. We focused on the CCM2 protein that might take place in blood vessel formation; we report here the expression patterns of CCM2 in prenatal development and adult human neocortex by means of immunohistochemistry and Western blot analysis. CCM2 was obviously detected in vascular endothelium and neuroglial precursor cells during development and also observed in arterial endothelium, neurons, some of the glial cells in adult neocortex. The expression patterns suggest that it could be one of the arterial markers whether this is a cause or a consequence of an altered vascular identity. CCM2 might play a role during vasculogenesis and angiogenesis during human brain development. Furthermore, with this study, CCM2 have been described for the first time in developing human neocortex.     

豚鼠早期实验性近视眼视网膜色素上皮细胞bFGF表达变化的研究

目的：研究豚鼠早期实验性近视眼视网膜色素上皮(retinal pigment epithelial,RPE)细胞前部及后极部碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的改变,探索近视的发病机制。

方法：两周龄豚鼠30只随机分为A组、B组、C组,每组10只,再随机选取5只10眼正常两周龄豚鼠不作任何干预,作为正常对照眼。选取任意眼戴一-10.00D凹透镜,分别饲养6、15、30d后除去镜片,验光及测眼轴长度确定近视形成后,采用细胞酶消化法培养豚鼠的前部及后极部RPE细胞,取3～6代RPE细胞进行免疫细胞化学、实时荧光定量PCR、Western-blot蛋白印迹法检查前部及后极部RPE细胞中bFGF表达变化。

结果：bFGF的表达定位于细胞浆和细胞核。免疫细胞化学、实时荧光定量PCR、Western-blot蛋白印迹法结果均表明：A、B、C三组实验眼和对照眼前部及后极部RPE细胞均有bFGF的表达,实验组前部及后极部与对照组相应前部及后极部比较,实验组中bFGF的阳性率低于对照组,差异有统计学意义(P<0.05); 而且,随着诱导时间的推移,实验组的阳性率逐渐降低,差异有统计学意义(P<0.05),对照组的阳性率不变,差异无统计学意义(P>0.05); 但实验组和对照组各组自身前部及后极部比较,差异无统计学意义(P>0.05)。

结论：实验组前部及后极部与对照组相应前部及后极部比较,bFGF的表达显著低于对照组。     

骨肉瘤常用化疗药物作用的分子机制及用药方法选择

目的 研究常用骨肉瘤化疗药物作用的分子机制,对临床用药方式提供指导.方法 将骨肉瘤细胞混悬液与不同浓度吡柔比星、顺铂、卡铂、甲氨蝶呤、异环磷酰胺分别作用24、48、72 h后,用倒置显微镜观察细胞生长情况.用MTT法测定每一孔的吸光度值OD,制作生长曲线,并求出药物的IC50.用IC50浓度的药物分别作用于骨肉瘤细胞24、48和72 h后用流式细胞仪进行细胞周期分析,了解细胞的周期分布及细胞的增殖活性.免疫印迹技术观察细胞的PCNA、Bcl-2、Bax、cyclin D1、cyclin E的表达.结果 在倒置显微镜下观察不同药物浓度作用于的骨肉瘤细胞,比较细胞死亡数量于时间关系.MTT实验:不同时间组及不同浓度组与对照组相比,对MG-63细胞的生长抑制作用差异均具有显著性(P＜0.05),呈现不同剂量-时间效应关系.细胞周期分析:在细胞各期可见明显凋亡峰,细胞周期各期所占比例无明显变化,抑制作用呈剂量依赖性.蛋白免疫印迹:PCNA在不同药物组骨肉瘤表达与对照组相比明显不同,不同药物组Bax表达与对照组明显增加,Bcl-2表达减少.cyclin D1、cyclin E的表达明显减少.结论 吡柔比星对MG-63细胞的抑制率和时间关系密切,和浓度有一定的关系.同样剂量吡柔比星5d的效果比3d好.卡铂具有与顺铂类似的效应,但卡铂如果替代顺铂在化疗中的作用,卡铂的量必须是顺铂的25倍以上.甲氨蝶呤作用于骨肉瘤细胞MG-63后,随着药物浓度的增加和时间的延长,作用逐渐增强,尤其是用量和效果有明显关系.异环磷酰胺在体外对骨肉瘤细胞无明显抑制作用.