大肠侧向发育型肿瘤细胞株双向电泳技术成功建立

目的:建立侧向发育型肿瘤细胞(LST)株双向电泳技术.方法:提取LST细胞株总蛋白质,采用不同pH范围IPG胶条进行LST细胞株总蛋白质双向电泳,并对实验步骤进行一系列的调整优化.结果:获得了较为清晰的LST细胞株双向电泳图谱,采用pH 3～10线性IPG胶条分离出1356±43个蛋白质斑点;pH 4～7 IPG胶条两种上样量(250μg和150μg)分别分离出了1285±51和989个蛋白质斑点.结论:通过相关条件的调整优化,成功建立了双向电泳技术,为研究LST特殊性生物学行为相关蛋白质奠定了坚实的基础.     

Familial homozygous hypercholesterolemia: clinical and cardiovascular features in 18 patients

Homozygous familial hypercholesterolemia (HFH) is a very rare autosomal dominant disease characterized by accelerated severe atherosclerosis. We examined 18 patients from 9 families with HFH. The age range was 6-30 years (mean = 16 years). Male to female ratio was equal. All patients had huge, multiple tuberous xanthomas on the skin and tendons. Mean +/- standard deviation of plasma cholesterol, triglycerides, low-density lipoproteins (LDL), and high-density lipoproteins (HDL) cholesterol levels were 608 +/- 89, 122 +/- 39, 550 +/- 88, and 26 +/- 8 mg/dl, respectively. Five patients (28%) had angina pectoris, two sustained a myocardial infarction, and one died at the age of 15 years. Two-dimensional echocardiography demonstrated supravalvular aortic stenosis in 3 of the 13 patients (23%). Coronary arteriography performed in 11 patients demonstrated significant obstruction in 6 patients, 2 each with single-, double-, and triple-vessel disease. Left main stenosis was present in 3 patients (27%). Supravalvular aortic narrowing was demonstrated in 6 patients (54%) and was associated with a gradient in 2 (25 and 35 mmHg, respectively). Segmental contraction abnormalities were detected in 2 of the 11 patients (18%). It is concluded that coronary artery disease is prevalent in patients with HFH and, based on the data presented, we recommend the performance of noninvasive technique, coronary arteriography and supravalvular aortography at an early age to detect and to follow the progression of the disease.     

L-肌肽对大鼠硒性白内障的影响

Objective To explore the effect of L-canosine in preventing and treating rat cataract induced by sodium selenite. Methods This was an experimental study. Cataract was induced in the rats by sodium selenite. Rats were divided into 3 groups and each group has 3 ones. L-canosine eye drops (50 g/L or 20 g/L) and 0.9% normal saline were respectively instilled to the rat eye for 3 weeks and examined. L-canosine was added to the medium of cultured rat cataract lens at 1.00, 0.10 and 0.01 g/L for 1 week and then examined. Cataract lens were studied by using two-dimensional eletrophoresis. Results One week after instillation of L-canosine, the scores of rat cataract were 2.22±0.65, 2.39±0.98 and 2.83±0.38 in 50 g/L, 20 g/L L-canosine and control groups, respectively. The lesion in 50 g/l, L-canosine group was lighter than that of the control group. There was significant difference between these two groups (P=0.013). On the 2nd and 3rd weeks, there was no difference among these three groups. After 1 week for culturing cataract lens, there was no difference between these three groups. The results of two-dimensional eletrophoresis showed that the protein number was 182 and 161 in the L-carnosine and control groups, respectively. High molecular weight protein decreased and low molecular weight protein increased in L-carnosine group. Conclusions L-carnosine at 50 g/L could restrain the development of early stage rat cataract. L-camosine could modulate the rat lens protein in vitro, but could not affect lens cataract scores.     

挫伤性屈光不正的眼超声生物学检测

对１１２例挫伤性屈光不正患者进行了超声生物学检测。结果提示，挫伤性近视患者眼球解剖结构存在两大变，即晶体前移和厚度增加，超声前房深度、晶本厚度在伤后的检测结果与对照组相比具显著性差异。并对挫伤屈光不正的发生机制进行了探讨。     

Polypeptide composition of normal and neoplastic human breast tissues and cells analyzed by two-dimensional gel electrophoresis

Summary The protein populations of epithelial cells cultured from two neoplastic and five non-neoplastic human breast tissues were resolved and displayed by two-dimensional polyacrylamide gel electrophoresis and silverstaining. With a computer-based image analysis system, we identified eight polypeptides which are present in both of the neoplastic cell lines, but absent from all five of the cultures of non-neoplastic breast cells. The eight polypeptides are not unique to cells cultured from neoplastic breast, because they are also found in cells cultured from non-breast tissues, both neoplastic and non-neoplastic. Two of the eight polypeptides ( M_r 25,000/pI 4.4 and M_r 31,000/pI 5.5) are present in the patterns of whole tissue samples from infiltrating ductal carcinomas and absent in most normal breast tissue.     

人晶状体上皮细胞蛋白质组的双向电泳和质谱鉴定

目的： 建立人晶状体上皮细胞蛋白质组研究体系，探讨双向电泳和质谱鉴定技术在人晶状体上皮细胞蛋白质组研究中的作用。 方法: 体外培养人晶状体上皮细胞株，用两种不同方法提取总蛋白，进行固相pH梯度(IPG)等电聚焦双向凝胶电泳，凝胶通过GS-800扫描仪（Bio-Rad）获取图像并使用PDQuest专业图像分析软件分析。在此基础之上，胰酶消化蛋白质斑点并进行质谱分析。 结果: 获得了重复性较好的人晶状体上皮细胞蛋白质组电泳图谱。晶状体蛋白质斑点在等电点pH值为4-7、相对分子质量为17-72 kD之间均有分布。其中高丰度蛋白点主要分布于分子量19-50 kD、PI 5-7范围内。2个蛋白点通过质谱分析和数据库的检索得到了初步的鉴定。 结论: 建立起了一个稳定的分析人晶状体上皮细胞蛋白质组学实验体系；为进一步研究人类晶状体在生理状态及白内障等病理条件下的改变提供了蛋白质组学的研究方法和途径。     

Biochemical characterization of I-Ak proteins from mutant antigen-presenting B-cell--B-lymphoma hybridomas

Chemically induced mutants of an I-Ak,d expressing antigen-presenting B-cell--B-lymphoma hybridoma have recently been generated by immunoselection in vitro and were found to possess alterations in some of their serologically and functionally defined I-Ak region dependent functions. In order to identify at the structural level the origin of the differences in serological and functional properties of these mutants, I-Ak molecules from several of these mutant hybridomas were compared biochemically to wild-type I-Ak polypeptides by two-dimensional gel electrophoresis and high-pressure liquid chromatographic tryptic peptide analyses. Two-dimensional gel electrophoresis indicated that no major structural alterations, resulting in changes in mol. wt or charge, had occurred in the Ak alpha or Ak beta polypeptides from the mutant cells. Likewise, Ak alpha peptide maps of the mutants were indistinguishable from the normal Ak alpha peptide maps. However, two of the three mutants studied did exhibit one additional peptide in their Ak beta peptide maps. These results suggest that the major deficiencies in T-cell-activating functions of these mutants are a result of a limited alteration in the Ak beta polypeptide primary structure.     

人胃黏膜组织蛋白质组二维电泳图像的获得

目的初步建立并优化人类胃黏膜组织蛋白质组分析所需的双向凝胶电泳技术，提高其分辨率及重复性。方法刮取手术胃黏膜组织，对以固相pH梯度为第一向的双向凝胶电泳的关键因素与环节，如样品处理、上样量、电泳参数、凝胶浓度和SDS凝胶电泳染色方法等进行一系列的优化。以固相pH梯度——IPG胶条(pH=3—10)进行第一向等电聚焦，以SDS均一胶(13％)的垂直电泳为第二向。结果成功地得到了胃黏膜组织的双向凝胶电泳图谱。     

皮肤角朊细胞与表皮干细胞双向电泳图谱的建立及差异分析

目的 建立皮肤角朊细胞与表皮干细胞的双向电泳图谱,并分析二者表达蛋白质的差异,为进一步研究体外调控表皮十细胞的增殖、分化提供线索.方法酶消化法获取单个表皮细胞悬液.Ⅳ型胶原快速贴壁法分选表皮干细胞.利用舣向电泳技术(2-DE)建立两种细胞的蛋白质表达图谱,并用Imagemaster 2D elite 5.0软件分析两种细胞的差异表达蛋一点.结果两种细胞的2-DE蛋白表达谱具有良好的重复性和可比性.皮肤角朊细胞与表皮干细胞的电泳图谱平均蛋白质点数分别为(982 ±18)个和(930 ±15)个,匹配点数为(850±13)个和(798±11)个,匹配率是86.56%和85.81%;两种细胞蛋白质间匹配点数是(886 ±8)个,匹配率是76.98%.找到差异蛋白点11个,其中只在表皮下细胞中表达或高表达的有8个;只在皮肤角朊细胞中表达或高表达的有3个.结论利用双向电泳法得到了分辨率较高儿重复性好的皮肤角朊细胞与表皮干细胞蛋白质组图谱,且二者存在有一定的差异.