Ubiquitin and heat shock protein expression in amyotrophic lateral sclerosis

The expression of two heat shock proteins, HSP72 and p57, in addition to ubiquitin, has been studied immunocytochemically in nine amyotrophic lateral sclerosis (ALS) cases and 10 age-matched controls. HSP72 and p57 antibodies did not identify the characteristic ubiquitin-immunoreactive inclusions present in anterior horn cells in ALS spinal cord. Antibodies to HSP72, but not to p57 or ubiquitin, strongly labelled structures corresponding to polyglucosan bodies in spinal grey matter. Such immunoreactive profiles were more abundant in ALS cases, although they were also present in control material. They were sometimes identified by haematoxylin and eosin and periodic acid Schiff reaction, but were not labeled by phosphotungstic acid haematoxylin or by antibodies to glial fibrillary acidic protein. Although ubiquitin, HSP72 and p57 are stress-induced proteins, they are expressed differently and might therefore have different significance in neuronal degeneration.     

肥胖患者血清C-反应蛋白水平变化与代谢综合征的关系

目的　比较肥胖患者急性期标志 -血清C 反应蛋白 (CRP)水平的变化并探讨与代谢综合征的关系。方法　选择 15 3例肥胖患者和 4 7例正常体重者 ,测定血清CRP水平 ,同时检测体重指数 (BMI)、腰围 (W )、腰臀比 (WHR)、血压、糖脂代谢参数、空腹胰岛素 (FIN)、胰岛素原 (PI)、胰岛素原 /胰岛素比值 (P/I) ,并对导致血清CRP改变的因素进行相关分析研究。结果　 ( 1)肥胖组血清CRP高于体重正常组 (P <0 .0 0 1) ,单纯肥胖组与肥胖伴糖代谢异常组之间无明显差异 (P =0 .4 13 ) ;( 2 )肥胖组相关分析显示血清CRP与W、WHR、SBP、TG、PI、HOMA IR、P/I呈正相关 ;( 3 )矫正年龄和BMI后 ,血清CRP与SBP、WHR和RI仍存在相关性 ,与其他变量的相关性消失。结论　 1、肥胖患者血清CRP水平明显高于非肥胖者 ,与SBP、TG、HOMA IR呈正相关 ,提示肥胖患者血清CRP水平升高与代谢综合征、胰岛素抵抗有密切关系 ;( 2 )血清CRP水平与P/I也呈正相关 ,提示血清CRP升高与 β 细胞功能缺陷可能也有一定关系 ;( 3 )BMI、WHR、SBP和PI是血清CRP升高的主要相关因素     

胞嘧啶脱氨酶的原核表达和多克隆抗体的制备

目的：在原核系统中表达并纯化大肠杆菌胞嘧啶脱氨酶（cytosine deaminase,CD)，制备鼠抗大肠杆菌CD多克隆抗体，方法：亚克隆CD基因到原核表达载体pMAL-c2和pBV222中，并转化入大肠杆菌DH5α内，诱导表达并纯化MBP-CD和6his-CD融合蛋白，用纯化的MBP-CD融合蛋白免疫小鼠制备多克隆抗体。结果：通过重组质粒的酶切筛选出重组阳性克隆，成功地表达和纯化出MBP-CD和6his-CD融合蛋白，用纯化的MBP-CD成功制备了鼠抗CD多克隆抗体，并用6his-CD和GST-CD重组蛋白进行Western印迹分析，证实了抗体的正确性，结论：应用多克隆抗体可以检测体内外CD基因的表达，为临床前和临床上深入开展CD基因的生物治疗研究提供重要的实验材料。     

Folding and function of the myelin proteins from primary sequence data.

To explain how the myelin proteins are involved in the organization and function of the myelin sheath requires knowing their molecular structures. Except for P2 basic protein of PNS myelin, however, their structures are not yet known. As an aid to predicting their molecular folding and possible functions, we have developed a FORTRAN program to analyze the primary sequence data for proteins, and have applied this to the myelin proteins in particular. In this program, propensities for the secondary structure conformations as well as physical-chemical parameters are assigned to the amino acids and the pattern of these parameters is examined by calculating their average values, autocorrelation functions and Fourier transforms. To compare two proteins, their sequences are aligned using a unitary scoring matrix, and homologies are searched by plotting a two-dimensional map of the correlation coefficients. Comparison of the corresponding myelin basic proteins (MBP) and P0 glycoproteins (P0) for rodent and shark showed that the conserved residues included most of the amino acids which were predicted to form the alpha or beta conformations, while the altered residues were mainly in the hydrophilic and turn or coil regions. In both rodent and shark the putative extracellular domain of P0 glycoprotein displayed consecutive peaks of beta propensity similar to that for the immunoglobulins, while the cytoplasmic domain showed alpha-beta-alpha folding. To trace the immunoglobulin fold along the P0 sequence, we compared the beta propensity curve of P0 with that of the immunoglobulin M603, whose three-dimensional structure has been determined. We propose that the flat beta-sheets of P0 are orientated parallel to the membrane surface to facilitate their homotypic interaction in the extracellular space. An extra beta-fold in the extracellular domain of shark P0 compared with rodent P0 was found, and this may result in a greater attraction between the apposed extracellular surfaces and may account for a smaller extracellular space as measured by x-ray diffraction. A computer search of the myelin protein sequences for functional motifs revealed sites for N-glycosylation, phosphorylation, nucleotide binding, and certain enzyme activities. We note especially that there are potential nucleotide binding sites in proteolipid protein (PLP), MBP and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP). This is consistent with the experimental observations that PLP acts like an ionophore or proton channel when reconstituted into planar lipid bilayers, MBP binds GTP, and CNP catalyzes in vitro the hydrolysis of 2',3'-nucleotides into corresponding 2'-nucleotides.     

牛骨形态发生蛋白的提取及异位诱导成骨的研究

本研究从1kg牛长骨中提取出部分纯化的牛骨形态发生蛋白(bBMP)160mg.电泳显示其主要区带分子量在33.7～13.4ku(34.0～13.5kd)之间,包括bBMP的生物活性成份18.1ku(18.2kd)蛋白质。将bBMP植入NIH小鼠肌肉内,5天诱导出软骨细胞,10天形成骨组织,14天出现骨髓。软骨或骨的诱发率87.5%。作者观察到bBMP诱导成骨的方式是软骨内成骨,其过程可分为间质细胞增生期、软骨细胞演变期和骨形成期。     

全膀胱切除回肠膀胱术15年总结(附196例报告)

目的　评价全膀胱切除治疗膀胱肿瘤的疗效及回肠膀胱术的远期效果。　方法　回顾性分析 1985年 1月至 2 0 0 0年 1月膀胱肿瘤行全膀胱切除回肠膀胱术 196例的临床资料。　结果　膀胱肿瘤累及膀胱颈部或膀胱三角区者 12 6例 (6 4.3 % ) ,浸润性膀胱癌 145例 (74.0 % ) ,移行细胞癌183例 (93.4% )。术后发生近期并发症 19例 (9.7% ) ,远期并发症 10例 (7.4% )。 135例随访 1～ 15年 ,平均 6 .6年 ,5年生存率 6 6 .2 %。　结论　全膀胱切除是浸润性膀胱癌首选治疗方法 ,回肠膀胱术简单易行 ,长期随访显示其并发症少 ,疗效确切 ,仍是一种较为理想的尿流改道方式。     

全去带乙状结肠原位可控膀胱术25例随访报告

目的：探讨全去带乙状结肠原位可控膀胱术的临床效果。方法：对25例施行该术式术后患者的可控性、尿动力、血清电解质、肾功能、生活质量等进行随访观察。结果：25例随访2－15个月，平均11．3个月。白天完全自控排尿25例（100％）；夜间完全自控排尿24例（96．0％），1例偶有遗尿；贮尿囊容量220－370ml，平均320ml；贮尿囊内压力1．86－3．92kPa，平均2.44kPa。IVU及贮尿囊造影示单侧贮尿囊输尿管反流2例（8．0％），无肾积水；膀胱镜检查2例（8．0％）贮尿囊内出现细沙样结石，1例（4．0％）贮尿囊后尿道吻合口狭窄，经尿道直视内切开术治愈。15例（60．0％）男性患者保留性功能，肾功能正常，1例（4．0％）出现一过性高氯血症。结论：全去带乙状结肠原位可控膀胱术手术成功率高，疗效可靠，患者生活质量高，是一种较理想的尿路分流术，值得推广。