人血清载脂蛋白A-I的分离、纯化及其测定

用超离心法制备人血清高密度脂蛋白(HDL);甲醇/氯仿脱脂;经Sephadex G-150层析及DEAE-纤维素(DE-52)离子交换得纯化载脂蛋白A-I(Apo A-I),免疫新西兰兔,获得效价为1:32的特异性抗人Apo A-I抗血清。用本室建立的单向火箭免疫电泳法测定了158名健康中年人与9名老人的血清Apo A—I含量,结果发现:β-脂蛋白<5g/L,总胆固醇<2mmol/L的人血清Apo A-I含量男女分别为2.06±0.16g/L和2.07±0.15g/L;而β脂蛋白≥5g/L,总胆固醇≥2mmol/L的人血清Apo A-I含量男女分别为1.75±0.15g/L和1.82±0.1g/L,由此可见,血脂正常的男性和女性的Apo A—I含量均显著高于血脂高的男性和女性。80岁以上老年人的血清Apo A-I含量为2.1±0.1g/L,也在正常范围。与文献报道比较,我国人的血清Apo A-I含量高于外国人,这与我国人HDL水平较外国人高一致。     

老年人血清胆红素水平与冠状动脉病变程度的关系

目的观察老年人血清胆红素与冠状动脉病变的关系。方法经冠状动脉造影确诊的冠心病患者80例,其中冠状动脉轻度狭窄组15例,中度狭窄组32例,重度狭窄组33例;另外选取冠状动脉造影正常者40例做对照。采用重氮苯磺酸法测定血清胆红素水平,并对胆红素与各因素进行直线相关分析。结果冠心病组血清胆红素水平明显低于正常对照组,血清胆红素水平与冠状动脉病变程度成显著负相关关系(r=-0.26,P<0.05)。结论冠心病患者血清胆红素水平较正常人明显降低。低血清胆红素可能是冠心病的一种危险因素。血清胆红素水平与冠状动脉狭窄程度有明显相关性。     

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.     

大鼠气管上皮细胞体内—体外转化模型的研究

本研究建立了大鼠气管上皮细胞体内－体外转化模型，大鼠气管内滴注苯并芘，三天后处死大鼠，消化气管上皮细胞，接种于无血清完全培养基。细胞形成集落后，换为选择培养基继续培养五周，统计转化率。结果显示，２５ｍｇ／ｋｇ和５０ｍｇ／ｋｇ的苯并芘可诱导大鼠气管上皮细胞转化及微核增加，用同样方法研究了煤焦沥青提取物，结果表明，剂量为８ｍｇ／ｋｇ和２５ｍｇ／ｋｇ的煤焦沥青提取物能明显诱导大鼠气管上皮细胞转化。     

庆大霉素经乳汁的排泄

采用荧光偏振免疫法测定１４例剖宫产术后应用肌注或静点庆大霉素的产妇乳汁及血清中庆大霉素的浓度，结果表明肌注或静点庆大霉素后３￣５小时，乳汁中浓度达峰值，血清中庆大霉素的浓度于给药后０．５小时达峰值。提示：无论肌注或静脉给庆大霉素均可经乳汁排泄，乳母不宜应用庆大霉素，即使产后已应用庆大霉素，应停药１３小时后方可哺乳。     

血清磷、肌酸激酶及其同工酶的测定在肠梗阻诊断中的应用

肠梗阻是临床常见的急腹症，绞窄性肠梗阻的死亡率可高达４５％～３０％，我们自１９９５年８月至１９９７年４月共收治肠梗阻５３例，其中单纯性肠梗阻２８例，经手术证实为绞窄性肠梗阻２５例（其中肠坏死８例），在治疗前均测定血清磷、肌酸激酶及其同工酶，其结果：本组病例中血清磷升高者１４例，其中以８例肠坏死为显著，肌酸激酶及其同功酶的升高也以绞窄性肠梗阻为明显，其中又以发病时间在４８小时以内者和８例肠坏死者为著，因此，我们认为：血清磷、肌酸激酶及其同工酶的测定可以作为判断肠管是否发生绞窄及坏死的辅助诊断指标     

银屑病患者血清促进皮肤成纤维细胞增生的实验研究

进行期寻常型银屑病患者混合血清以5%的比例加入培养基中,作用于体外培养的正常人皮肤成纤维细胞.结果发现,对该细胞的增生具有明显的促进作用.提示银屑病不仅是表皮细胞的异常增生,真皮成纤维细胞的异常和系统性因素在其病理生理方面也占有重要地位.     

Sialic acid concentrations in the urine of men with and without renal stones

Summary It has been suggested that urinary sialidase may play a role in the formation of renal stones. The present study was therefore undertaken to compare spectrophotometrically the different types of sialic acid concentrations and sialidase activities in fresh first morning urine specimens of men (21–65 years) with (13) and without (9) calcium oxalate renal stones. Although the free urinary sialic acid concentrations of the two groups of men were statistically about the same (P=0.0614), the total (P=0.003) and bound (P=0.0012) urinary sialic acid concentrations differed significantly. Both the total and bound sialic acid concentrations were lower in the urine specimens of the stone patients than in their healthy counterparts. This decrease in urinary sialic acid concentrations was firstly thought to be the result of elevated breakdown enzymes of sialic acid, which would favour the production of pyruvate. However, spectrophotometric determinations of the endogenous pyruvate concentrations of the two types of urine specimens did not differ significantly (P=0.0708). Secondly, the decrease in total urinary total sialic acid concentration of stone patients, could be attributed to less sialic acid synthesis or less renal excretion. Therefore, the same experiments were repeated using serum of 13 patients and 9 healthy men. Conversely, the total (P=0.4425) and bound (P=0.2850) serum sialic acid concentrations were found to be similar in the two types of subjects. However, the free serum sialic acid concentration of stone patients was significantly lower than in the healthy subjects (P=0.0062). This phenomenon is also reflected in the average ratio for serum free: bound sialic acid in healthy and stone patients, 1:7.9 and 1:18.7 respectively (P=0.0009). The lower free serum sialic acid concentration may lead to lower renal excretions of sialic acid. This may explain the decrease in total urinary sialic acid concentration in stone patients. The lower bound urinary sialic acid concentrations in patients was also reflected in the urinary free: bound sialic acid ratio for healthy (1:2.3) and stone patients (1:1.3). The difference between these two groups of men was highly significant (P=0.0001). This phenomenon might be explained by the urinary sialidase activities, which was spectrophotometrically determined at 334 nm at 37°C of 11 patients with stones and 17 healthy men. The ages of both groups of men were the same (P=0.326). An increase in urinary sialidase activity was observed with the stone patients (P=0.00001) when compared to specimens of healthy men. This might explain the decrease in urinary bound sialic acid concentration of the stone group. It seems from these results that the urinary concentration of sialic acid and the activity of urinary sialidase, may play a role in the pathogenesis of the multifactorial disease, urolithiasis.     

~(125)碘-绒毛膜促性腺激素的电泳纯化及分析

标记抗原在贮存过程中,由于受幅射分解和脱碘影响而使免疫活性和生物活性降低。我们对自制的~(125)碘-绒毛膜促性腺激素进行了实验观察。结果显示:贮存1月后,其免疫活性和生物活性分别降低了33.1％和44.0％;经电泳纯化后的标记物的免疫活性和生物活性明显提高,分别达到了最初的90.5％和95.3％。     

质粒pGH/BL_(21)在大肠杆菌中表达条件的研究

目的　寻找质粒 pGH/BL2 1在大肠杆菌中的最佳表达条件。方法　当它在大肠杆菌中表达时 ,考查不同IPTG诱导浓度及不同的诱导时间对表达产物的影响 ,表达产物的量以SDS -PAGE电泳方法分析。结果　该基因的最佳表达条件为 :IPTG诱导浓度 1.0mmol/L ,诱导时间 4h。结论　IPTG对不同的基因在不同的表达体系中需不同的诱导条件 ,只有找到它的最佳诱导条件才能保证终产物的最大产率