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。     

谷胱甘肽合成酶系基因协调表达体系的构建

通过 PCR获得 E.coli B谷胱甘肽合成酶系基因 ( gsh I,gsh II)片段 ,结合定点突变稀有起始密码子 ,设定 gsh I与 gsh II位置与距离 ,构建双顺反子重组表达载体 p Trc99A/gsh I- gsh II,建立GSHI、GSH- II蛋白表达体系。结果表明 :以 0 .0 8mmol/L IPTG于 2 8℃诱导工程菌 E.coli BL2 1( DE3) ( p Trc99A/gsh I- gsh II) ,GSH- I、GSH- II蛋白比为 4.5∶ 1 ( m∶ m)时谷胱甘肽合成能力最高 ,达到每克湿菌体 8.5 mg。通过构建单顺反子重组表达载体 p Trc99A/gsh I、p Trc99A/gsh II测定GSH- I与 GSH- II蛋白的最适配比 ,结果表明 :在 GSH- I、GSH- II蛋白总量恒定的情况下 ,要提高谷胱甘肽产率 ,两者比例以 3∶ 1～ 6∶ 1为宜     

激素替代治疗对绝经后Ⅱ型糖尿病和高血压患者肾脏微血管病变的影响

目的 :探讨激素替代治疗对绝经后Ⅱ型糖尿病和高血压患者肾脏微血管病变的影响。方法 :36例患有Ⅱ型糖尿病和高血压的绝经后妇女随机分为治疗组和安慰剂组各 18例 ,采用双盲法予口服克龄蒙或安慰剂 1片 /日 ,共 4 5个周期 ,测量用药前后各参数值并进行比较及线性相关分析。结果 :治疗组用药前后 2 4小时尿蛋白定量由 ( 0 4 45± 0 0 36 )g降到 ( 0 36 1± 0 0 32 ) g(P <0 0 1) ,内生肌酐清除率由 ( 92 0± 5 2 )ml/min增到 ( 99 1± 4 8)ml/min (P <0 0 5) ,空腹血糖由 ( 7 0 2± 0 4 3)mmol/L降到 ( 6 55± 0 31)mmol/L(P <0 0 5) ,血清总胆固醇由 ( 6 6 6±0 2 8)mmol/L降到 ( 5 71± 0 71)mmol/L(P <0 0 1) ,血压无明显改变。 2 4小时尿蛋白定量和内生肌酐清除率与其他参数间无显著相关性。安慰剂组用药前后各项指标无明显改变。结论 :激素替代治疗对绝经后Ⅱ型糖尿病和高血压患者肾脏微血管病变有改善作用     

日本血吸虫成虫67kD蛋白的分离与鉴定

目的 :分离日本血吸虫感染及免疫血清识别的成虫抗原 (AWA)中的特异蛋白带 ,为血吸虫病免疫诊断提供新的抗原分子。方法 :免疫印迹法分析AWA的特异蛋白带 ,电泳层析法分离靶抗原。结果 :获得了感染血清和免疫血清识别的 6 7kD蛋白。结论 :电泳层析法是一种分离血吸虫抗原的有效方法     

MMP-9和TIMP-1表达失衡与乳腺癌浸润、转移的相关性

目的 研究乳腺癌组织中基质金属蛋白酶（MMP－9）和金属蛋白酶组织抑制因子（TIMP－1）的表达变化与乳腺癌生物学行为及淋巴结转移的关系。方法 应用SP免疫组织化学方法检测85例乳腺癌组织MMP－9，TIMP－1及细胞增殖核抗原ki-67的表达情况。结果 MMP-9阳性染色率90．59％，MMP－9阳性表达与肿瘤浸润，淋巴结转移，ki-67指数及TNM分期呈正相关（Pearson列联系数分别为P=0.03,P=0.02,P=0.004和P＝0．0000，P＜0．05，0．01。TIMP－1阳性染色率为78．82％，TIMP－1阳性表达与肿瘤浸润，淋巴结转移及TNM分期浸润，转移及ki-67指数显著相关（P＜0．05，P＜0．01，P＜0．001）。结论 MMP－9和TIMP－1表达失衡与乳腺癌浸润及淋巴结转移密切相关。