我国婴儿型Ⅰ型多囊肾病（EPKD—Ⅰ型）调查分析

目的：了解我国ＩＰＫＤ－Ⅰ型的流行病学特点及基本状况，为进一步研究及治疗此类畸形提供一些参考资料。方法：１９８７～１９９２年中国出生缺陷监测网以医院为基础在全国对孕２８周至产后７天的围产儿进行监测，对３７４例ＩＰＫＤ－Ⅰ型患儿进行流行病学分析。结果：我国ＩＰＫＤ－Ⅰ型的发生率为０．８３／万，发生率上降趋势，城乡、男女发生率均无显著性差异。４４．３８％的ＩＰＫＤ－Ⅰ型患儿为低体重儿，５０．２６％的Ｅ     

抗人纤维蛋白单克隆抗体重链和轻链可变区基因克隆和表达

目的：为了获得能与人纤维蛋白结合的最小的活性分子。方法：采用逆转录－多聚酶链反应技术从分泌抗人纤维蛋白单克隆抗体的８Ｅ５杂交瘤细胞中分离出免疫球蛋白重链和轻链可变区基因（ＶＨ和ＶＫ）。通过基因重组构建了ＶＨ－ＶＫ的表达载体ｐＯＰＥ５１－８Ｅ５。转化大肠杆菌ＪＭ１０９后用异丙基－β－Ｄ－硫代半乳糖苷诱导表达。结果和结论：经聚丙烯酰胺电泳和免疫杂交证明产物表达于细菌的壁膜间隙和包涵体中，表达产物为分子量３００００的单链抗体分子，具有特异性识别和结合人纤维蛋白的活性。表达产量为转化菌总蛋白的２８．９％。     

淀粉酶同工酶琼脂糖电泳测定法

淀粉酶同工酶琼脂糖电泳测定法罗玲，赵月霞，杨振华，李云霞，贾天军（北京医院检验科，北京１００７３０）α－淀粉酶（ＥＣ．３．２．１．１，Ａｍｙ）的测定一直用于诊断胰腺疾病。但α－Ａｍｙ除来自胰腺外．还来自唾液腺，即胰型锭粉酶（Ｐ－Ａｍｙ），唾液型淀粉酶...     

神经元特异烯醇化酶的纯化方法

从牛脑制备ＮＳＥ纯品。纯化步骤依次为硫酸铵分级沉淀，ＤＥＡＥ－Ｓｅｐｈａｃｅｌ离子交换层析，Ｓｅｐｈａｃｒｙｌ－ｓ－３００凝胶过滤层析和ＤＥＡＥ－Ｓｅｐｈａｄｅｘ Ａ－５０离子交换层析。所得ＮＳＥ纯品，ＰＡＧＥ呈单一区带，ＳＤＳ－ＰＡＧＥ亚基分子量４６ＫＤ，ＩＥＦ－ＰＡＧＥ ｐＩ４．７～４．９，同工酶电泳仅显示γγ酶活性区带，酶比活性３０．１Ｕ／ｍｇ蛋白，得率３８．６％。     

人重组sIL—6R的纯化与鉴定

采用免疫吸附亲和层析，将抗ｓＩＬ－６Ｒ的单克隆抗体交联于ＣＮＢｒ活化的Ｓｅｐｈａｒｏｓｅ４Ｂ。收集ｓＩＬ－６Ｒ转基因细胞的培养上清。经预处理后上亲和柱，获得高纯度的ｓＩＬ－６Ｒ重组蛋白。经ＳＤＳ－ＰＡＧＥ电泳后出现一条区带，分子量为５５ｋＤ。     

老年2型糖尿病患者与一氧化氮等氧自由基关系的变化

目的：研究老年２型糖尿病患与一氧化氮等氧自由基的关系,方法：检测５０例老年２型糖尿患（患组）和５０例健康人（对照组）血浆一氧化氮（Ｐ－ＮＯ）,维生素Ｃ（Ｐ－ＶＣ）,维生素Ｅ（Ｐ－ＶＥ）、β－胡萝卜素（Ｐ－β－ＣＡＲ）含量及红细胞超氧化物坡化酶（Ｅ－ＳＯＤ）,过氧化氢酶（Ｅ－ＣＡＴ）、谷胱甘肽过氧化物酶（Ｅ－ＧＳＨ－ＰＸ）活性。结果：与对照组比较,患组除Ｐ－ＮＯ值显升高（Ｐ〈０．０１）外,余各检测值均显降低（Ｐ均〈０．０１）,结论：老年２型糖尿病患ＮＯ代谢异常,氧化、过氧化损伤加剧。     

血小板衍生的内皮细胞生长因子

血小板衍生的内皮细胞生长因子是一种分子量为４５ＫＤａ的阴离子蛋白质，主要分布于人血小板和胎盘，具有Ｅ．ｃｏｌｉ的胸苷磷酸化酶化酶的活性，在体外可刺激内皮细胞有丝分裂和ＤＮＡ合成，在体内可促进血管的发生和生长，本文就ＰＤ－ＥＣＧＦ的提纯产生和分布，特性与分子结构，基因结构与定位以及生物学特性等问题进行综述。     

肠内营养支持对ICU患者细胞免疫的影响

目的：研究肠内营养（ＥＮ）支持对ＩＣＵ患者的细胞免疫功能的影响。方法：对１１例健康人和１２例ＩＣＵ患者经ＥＮ支持前后的细胞免疫功能进行检测。结果：ＩＣＵ患者的免疫功能明显低于健康,表现为自然杀伤（ＮＫ）细胞活性和Ｔ细胞亚群ＣＤ４,ＣＤ４／ＣＤ８明显降低。经一段时期的ＥＮ支持后,细胞免疫功能有回升,表现为ＣＤ３,ＣＤ４,ＣＤ４／ＣＤ８和ＮＫ细胞活性明显升高,白介素－α（ＩＬ－α）浓度及ＩＬ－２分泌细     

重组抗原在戊型肝炎病毒感染诊断中的应用

目的探讨重组抗原在戊型肝炎病毒（ＨＥＶ）感染诊断中的应用。方法用ＨＥＶ重组抗原建立酶免疫试验（ＥＩＡ）检测肝病患者和健康人群血清中抗戊型肝炎病毒抗体（抗－ＨＥＶ）。结果５５份用新加坡ＤＢＬ公司抗－ＨＥＶ诊断试剂盒检测为抗－ＨＥＶ阳性的肝炎患者血清中，５３份（９６．４％）用本方法检测也为阳性；４５份ＤＢＬ试剂盒检测为抗－ＨＥＶ阴性的血清中，本法检测有３份为抗－ＨＥＶ阳性。检测甲、乙、丙型肝炎患者血清各３０份，抗－ＨＥＶ阳性率分别为１３．３％（４／３０）、１３．３％（４／３０）和１０．０％（３／３０）。健康人抗－ＨＥＶ阳性率为５．０％（８／１６０）。重组抗原与合成肽抗原混合用于ＥＩＡ，可以检出两种抗原单独阳性的抗－ＨＥＶ。结论以重组抗原为基础的ＥＩＡ特异性强、灵敏度高、快速简便，是戊型肝炎血清学诊断及流行病学调查的一种可靠方法     

尿液中α-淀粉酶的提纯及性质研究

为了制备淀粉酶测定的参考品，从正常人尿液中提取了α－淀粉酶（Ａｍｙ）并对其特性进行了研究。该提制品淀粉酶的经活性达８４．３ｋＵ／ｇ蛋白，几乎不含其它杂酶（测不到ＡＬＴ、ＳＡＴ、ＬＤ、ＣＫ、ＧＧＴ、ＣｈＥ、ＡＬＰ活性浓度）。ＳＤＳ－聚丙烯酰胺凝胶电脉显示一条Ａｍｙ区带。其催化特性和人血清Ａｍｙ非常相似，用亚乙基封闭的Ｇ７－ＰＮＰ（ＥＰＳ）作底物，偶联多功能α－葡萄糖苷酶测定其米氏常数为０．１８９ｍｍ     

Characterization of amylases produced by tumors

Amylases were purified and characterized from three amylase-producing human tumors. The relative molecular mass of the amylases was estimated to be 54 000 on sodium dodecyl sulfate/polyacrylamide gel electrophoresis, different from human salivary amylase (61 000 and 64 000) and human pancreatic amylase (60 000). The tumor amylases had completely identical antigenicities with human salivary and pancreatic amylases against antibody to human salivary amylase, while the intensity of the tumor amylases was less than 20% of that of human salivary and pancreatic amylases on single radial immunodiffusion. On isoelectric focusing, two of the three tumor amylases showed a major peak at pH 6.4, which corresponded to a major peak of human salivary amylase, the other showed a major peak at pH 6.4, which corresponded to a minor peak of human salivary amylase. The three tumor amylases showed similar amino acid composition, different from those of human salivary and pancreatic amylases. These findings suggest that tumor amylases have a tertiary structure similar to that of normal human amylases, but differ from them in amino acid composition.     

Human amylase isoenzymes separated on concanavalin A--Sepharose.

Human salivary amylase and pancreatic amylase were purified and characterized. These amylases gave two bands and one band, respectively, each staining for both protein and sugar, after electrophoresis on sodium dodecyl sulfate--polyacrylamide gel. The relative molecular mass (Mr) or pancreatic amylase was calculated to be 60 000; for the two components (A and B) of salivary amylase the Mr were 61 000 and 64 000. The two salivary amylases were separated by chromatography on concanavalin A--Sepharose; only component B bound to concanavalin A. The carbohydrate content of pancreatic amylase was 1.61 +/- 1.02% (SD), and of salivary amylases A and B 2. 18 +/- 0.71% and 8.77 +/- 2.28%, respectively. The salivary and pancreatic amylases had completely identical antigenicities against antibody to either. On isoelectric focusing, pancreatic amylase showed one peak at pH 7.0, salivary amylase A showed a major peak at pH 6.4 WITH A TRACE OF MATERIAL At pH 5.9, and salivary amylase B a major peak at pH 5.9 and one minor peak at pH 6.4. Serum amylase was separated into two major peaks with isoelectric points (pl) of 6.4 and 7.0, respectively, and one minor peak, with a pl of 5.9. Only a small part of the serum amylase with a pl of 5.9 combined with concanavalin A; the two other serum amylases did not.     

Immunoreactivities of alpha-amylase of humans and rats.

The immunoreactivities of amylase from human saliva and pancreatic juice and rat parotid and pancreas were investigated. Antisera were prepared in rabbits against each of the human and rat amylase. Human salivary and pancreatic amylases reacted similarly with the antibodies to both human salivary and pancreatic amylases. Rat parotid and pancreatic amylases reacted differently with the antibodies to both rat parotid and pancreatic amylases.     

Suitability of control materials in the differential inhibition assay for human pancreatic and salivary amylase

We investigated the behavior of 26 quality-control sera with the inhibitor method for differential amylase (EC 3.2.1.1) assay. We also studied the sensitivity to the wheat-derived inhibitor of pancreatic amylases from 10 different animals in comparison with human pancreatic and salivary amylase. The results indicate that only control materials containing human amylases can be measured accurately. The animal amylases (bovine, equine, porcine) used in many quality control sera are relatively insensitive to the inhibitor as compared with human pancreatic and salivary amylase.     

Use of a salivary alpha-amylase inhibitor in the saliva biochemical study

Amylolytic activity of oral fluid, saliva, and serum is maintained by alpha-amylases of different origin. Salivary and nonsalivary (mainly pancreatic) amylases can be recognized by reliable and simple in vitro tests with human salivary alpha-amylase inhibitor.     

Amylolysis of a chromogenic substrate, Cibachron Blue F3GA-amylose: kinetics and mechanism

We compared the modes of action of human pancreatic, human salivary, and porcine pancreatic amylases on Cibachron Blue F3GA-amylose. Both human enzymes showed similar catalytic activity with almost equal Vmax but dissimilar apparent Km's. The ratios of soluble dyed oligosaccharides to reducing substances were identical. Porcine pancreatic amylase exhibited less than half the Vmax of the human enzymes and a smaller apparent Km. Reducing substances were formed faster than were the soluble dyed products. These differences in amylolytic action can be explained by differences in the degree of the "multiple attack" mechanism. Introduction of dye substituents into the amylose molecule did not alter the substrate characteristics of amylose toward human serum amylase.     

Agarose electrophoresis and inhibitor tests for isoamylase determination can give complementary clinical information

We report the presence of an extremely high proportion of "aged" amylase in the serum and cyst fluid of a patient with a pancreatic pseudocyst. A salivary amylase inhibitor test helped us to differentiate these "aged" pancreatic amylases from salivary fractions having a similar electrophoretic mobility.     

Rapid quantitative, specific measurement of pancreatic amylase in serum with use of a monoclonal antibody

In this rapid quantitative assay for pancreatic alpha-amylase (EC 3.2.1.1) in serum, we precipitate salivary amylases by 10-min incubation with monoclonal anti-salivary amylase antibody immobilized on particles of polyvinylidene fluoride. We then centrifuge the serum mixture and measure the pancreatic amylase activity remaining in the supernate by a kinetic method. The assay requires 50 microL of serum and the standard curve is linear to at least 1300 U of pancreatic amylase per liter of serum. CVs were 1.3% within-run, 6-8% day-to-day. Apparent analytical recovery of pancreatic amylase activity added to serum was 101% +/- 2%. Addition of purified salivary amylase, 356 U/L, to sera gave a value for apparent pancreatic amylase of less than 4 U/L, or 1% of the added salivary amylase activity. This assay correlated well with an electrophoretic method (slope, 0.97-0.99; intercept, 0.5 to -4 U/L; correlation coefficient, 0.946-0.990; and standard error of the estimate 3-5 U/L). Estimated normal reference intervals with maltotetraose as substrate were: total amylase, 39-118 U/L; pancreatic amylase, 11-50 U/L; and salivary amylase, 18-79 U/L.     

Characterization of two phospholipases A2 in serum of patients with sepsis and acute pancreatitis.

Pancreatic phospholipase A2 and non-pancreatic ascitic phospholipases A2 were studied in sera of healthy individuals and of patients suffering from sepsis or acute pancreatitis. In gel filtration experiments, immunoreactive ascitic phospholipase A2, as determined in serum by a time-resolved fluoroimmunoassay, eluted either unassociated with an apparent M(r) of 10,000-14,000 or associated with proteins of high molecular mass. Catalytically active ascitic phospholipase A2 was associated with high molecular weight proteins. In acute pancreatitis the catalytically active and immunoreactive pancreatic phospholipase A2 eluted mainly as a protein of M(r) of 14,000. The results of the gel filtration experiments indicate that pancreatic phospholipase A2 is not associated with other proteins in human serum, whereas ascitic phospholipase A2 is associated with protein(s) of relative high molecular weight, or exists in different polymeric forms. We also purified phospholipase A2 from sera of healthy individuals by ion exchange chromatography and HPLC. The enzyme was homogenous, displayed an M(r) of approximately 13,500 as judged by SDS-polyacrylamide gel electrophoresis, and reacted with an antibody raised against ascitic phospholipase A2.