电泳法测定碱性磷酸酶同工酶的临床意义

目的　探讨碱性磷酸酶同工酶测定的临床意义。方法　用神经氨酸酶孵育血清快速电泳法检测 181例血清ALP同工酶。结果　健康成人ALP同工酶电泳主要为肝型和骨型 ,其所占比例分别为 34.5 %～ 6 2 .3%和 37.7%～6 5 .5 % .男女性别之间ALP无明显差异 (P >0 .0 5 ) .儿童ALP同工酶主要为骨型ALP ,占 85 %以上 .肝、骨ALP分离彻底 ,区带清晰。肝病 :主要为肝型ALP ,达 80 %以上 ;骨病 :主要为骨型ALP ,达 80 %以上 ;阻塞性黄疸 :为肝型和骨型ALP活性均增高 ,但以肝型ALP增高更为突出 ;佝偻病 :为骨型ALP轻、中度升高 ,不超过总活性的 70 % ;孕妇 :在肝和骨之间出现胎盘型ALP峰。结论　测定ALP总活性及其同工酶对引起ALP活性增高疾病的诊断和鉴别诊断有重要的临床意义。     

碱性磷酸酶同工酶在老年女性骨质疏松诊断中的应用

目的 探讨碱性磷酸酶（ALP）同工酶在老年女性骨质疏松诊断中的应用。方法 采用神经氨酸苷酶,用琼脂糖电泳法对体检正常者50例（对照组）和老年女性骨质疏松患者55例（骨质疏松组）血清中的ALP同工酶进行分离分析。结果 经神经氨酸苷酶处理后,琼脂糖电泳法可以分离肝和骨ALP;两组肝型ALP比较,差异无显著性意义（P〉0.05）;骨型ALP比较,差异有显著性意义（P〈0.05）;热失活法和电泳法检测骨ALP具有显著的相关关系（r=0.92,P〈0.05）。结论 骨型ALP可为临床诊断老年女性骨质疏松提供可靠的依据。     

未成年人血清碱性磷酸酶活性的生理变化

人血清碱性磷酸酶(ALP)活性测定可用于骨病、肝胆疾患等的辅助诊断和鉴别诊断。由于未成年人骨骼处在生长阶段,其骨型ALP同工酶活性显著升高,导致血清ALP活性高于成年人。我们在实     

改良琼脂糖电泳法测定BALP在前列腺癌骨转移诊断中的应用

目的探讨改良琼脂糖电泳法检测骨碱性磷酸酶同工酶(BALP)在前列腺癌(PCa)骨转移诊断中的应用。方法神经氨酸苷酶处理血清,采用改良的琼脂糖电泳法分离测定50例健康体检者和163例PCa患者血清中的ALP同工酶。结果 (1)普通的琼脂糖电泳法对肝和BALP的分辨率较低,改良琼脂糖电泳法提高了分辨率。(2)PCa组的总ALP和BALP均高于对照组,PCa骨转移组的总ALP和BALP活性均显著高于对照组和无骨转移组(P0.05),PCa无骨转移组的总ALP和BALP活性和对照组差异无统计学意义(P0.05)。结论 BALP与PCa骨转移存在一定的相关性,经神经氨酸苷酶处理血清后,采用改良琼脂糖电泳法分离ALP同工酶有助于临床辅助诊断PCa有无骨转移。     

改良琼脂糖电泳法分离碱性磷酸酶同工酶及其在肾脏疾病诊断中的应用

目的　用改良琼脂糖电泳法分离碱性磷酸酶同工酶并探讨在肾脏疾病诊断中的应用。方法　用改良琼脂糖电泳法对正常体检者和 16 2例肾脏疾病患者碱性磷酸酶 (ALP)同工酶进行分离和定量分析。结果　发现在慢性肾衰患者血清中存在胆汁型ALP ;肾小球肾炎和肾盂肾炎的ALP同工酶活性与正常者无显著性差异 ,慢性肾衰的ALP同工酶活性则显著高于正常体检者。在 6 9例慢性肾衰患者中 ,有 2 4例出现胆汁型ALP ,阳性率为 34 8%。结论　改良琼脂糖电泳法能定量分析肝、骨、胆汁等ALP同工酶 ,适合常规实验室应用     

碱性磷酸酶同工酶在肿瘤转移诊断中的应用

目的 探讨碱性磷酸酶(ALP)同工酶与肿瘤转移的相关性.方法 用琼脂糖电泳法对50例体检健康者和134例恶性肿瘤患者血清中的ALP同工酶进行分析.结果 肿瘤组的总ALP和同工酶活性均明显高于健康对照组;肿瘤组出现了胆汁型ALP,而健康对照组未出现,肿瘤组中有肝转移的胆汁型ALP活性高于其他各组;肿瘤组的骨型ALP活性高于健康对照组,而肿瘤组中有骨转移的骨型ALP活性高于其他各组.结论 胆汁型ALP与肿瘤肝转移有一定的相关性,骨型ALP与肿瘤骨转移有一定的相关性.     

肝型及骨型碱性磷酸酶的简易定量分离法

肝型和骨型碱性磷酸酶(ALP)的电泳迁移率很近似,用各种电泳方法都难以将两者分开。虽然它们的耐热性有所不同,但当两者混合一起或与其它同工酶共同存在时,仍不易鉴别。作者使用敏感度较好而且操作简便的醋酸纤维板(Titan-111)电泳,测定标本加热前后的ALP活性并算出加热后的活性残存率,根据两种方法的结果建立联立方程式,可以分别算出肝型及骨型ALP所占的活性比值。将血清标本在56℃加热10分钟,加热前后用King-King氏法测定ALP总活性。标本在加热前后还要分别进行电泳;使用醋酸纤维板为支持物,并用pH8.8及离子强度0.075的Tris-巴比妥缓冲液,4℃,180V,30分钟。电泳完毕后用5ml基质染色液〔内含α-萘酚ASXM磷酸钠盐2.5mg,0.1M MgCl,     

碱性磷酸酶同工酶对原发性胆汁性肝硬化的诊断价值初探

目的初步探讨血清碱性磷酸酶(ALP)同工酶对原发性胆汁性肝硬化(PBC)的辅助诊断价值.方法用Sebia琼脂糖凝胶电泳分离30名正常人、23例PBC患者和25例其他肝胆疾病患者的血清ALP同工酶,经光密度扫描仪扫描后,分析各型同工酶的相对含量.结果血清ALP同工酶经电泳后可分为肝型、胆汁型、肠型和骨型4个区带.PBC患者的肝型ALP的相对含量最高,约为56%～70%,显著高于正常对照及乙肝后肝硬化患者(P＜0.05),而其胆汁型ALP的相对含量显著低于肝外阻塞性黄疸患者(P＜0.05).结论ALP同工酶分析可辅助诊断PBC.     

碱性磷酸酶同工酶对原发性胆汁性肝硬化的诊断价值初探

目的　初步探讨血清碱性磷酸酶(ALP)同工酶对原发性胆汁性肝硬化(PBC)的辅助诊断价值。方法 　用Sebia琼脂糖凝胶电泳分离30名正常人、23例PBC患者和25例其他肝胆疾病患者的血清ALP同工酶,经 光密度扫描仪扫描后,分析各型同工酶的相对含量。结果　血清ALP同工酶经电泳后可分为肝型、胆汁型、肠 型和骨型4个区带。PBC患者的肝型ALP的相对含量最高,约为56%～70%,显著高于正常对照及乙肝后肝硬 化患者(P<0.05),而其胆汁型ALP的相对含量显著低于肝外阻塞性黄疸患者(P<0.05)。结论　ALP同工酶 分析可辅助诊断PBC。     

胆总管囊肿患儿血清肝型碱性磷酸酶同工酶的分析

目的分析胆总管囊肿患儿血清碱性磷酸酶（ALP）同工酶[肝型ALP（L-ALP），特别是L2-ALP]的临床价值。方法用Sebia琼脂糖凝胶电泳分离25名胆总管囊肿患儿和48名对照儿童的血清ALP同工酶，经光密度扫描仪扫描后，分析各型同工酶的相对含量。结果血清ALP同工酶经电泳后可分为肝型（L1和L2）、胎盘型（P1和P2）、肠型（I1、I2、I3）和骨型（B）4个部分。通过这种方法，L-ALP和B-ALP得到了很好的分离。儿童血清中L2-ALP的水平与胆总管囊肿的发生具有明显的相关性（P〈0．01）；胆总管囊肿患儿术后L2-ALP水平显著降低。结论胆总管囊肿患儿血清L-ALP同工酶的测定具有一定的临床价值。     

Alkaline phosphatase isoenzyme patterns in malignant disease.

Early treatment of patients with malignant disease and liver or bone metastasis may increase their survival time. We have used the activity patterns of liver and bone isoenzymes of alkaline phosphatase (ALP), separated by agarose gel electrophoresis, to detect early metastasis. We studied ALP isoenzyme patterns in a background population of 101 patients with no evidence of any disease that might influence this pattern; a healthy reference population (n = 330); and the following three groups of patients: 143 with malignant disease, 47 with nonmalignant liver disease, and 22 with nonmalignant bone disease. Cutoff and predictive values of liver ALP, high-molecular-mass (high-M(r)) ALP, and bone ALP were established for detecting liver and bone metastasis. The positive predictive value of liver and high-M(r) ALP was higher than that of total ALP in detecting liver metastasis, but liver and high-M(r) ALP did not enable us to differentiate between malignant and nonmalignant liver disease. Total ALP activity was of slightly more value than liver and high-M(r) ALP in enabling us to rule out liver metastasis. From bone ALP activity we could not distinguish between nonmalignant bone disease and bone metastasis. The negative predictive value of bone ALP in the diagnosis of bone metastasis was low, but its positive predictive value was high and superior to that of total ALP.     

Cellulose acetate electrophoretic determination of bone alkaline phosphatase activity in healthy subjects and diabetic patients with and without osteopenia.

We adapted the electrophoretic method of bone alkaline phosphatase (ALP) determination using neuraminidase from Vibrio cholerae to separate bone and liver ALP on cellulose acetate membrane. Treatment of separator plus serum (1:8, neuraminidase 111 U/l in final) for 10 min at room temperature (25 +/- 1 degree C) and subsequent electrophoresis made it possible to quantify bone ALP activity simply and rapidly. The precision of the data was at the level of CV of 1.6% (within-day) and 4.7% (day-to-day), with recovery rates of 97-103%. The normal range of bone ALP activity depended on age and sex. Seventy-eight diabetes mellitus (DM) patients, excluding those with renal failure, were divided into two groups of those with and without osteopenia with matching of age (+/- 3 years) and sex. Bone ALP (P < 0.001) and total ALP (P < 0.05) activities and urine calcium/creatinine ratio (P < 0.05) were significantly higher in DM with osteopenia than in DM without osteopenia. Therefore, bone formation and absorption may be accelerated in DM with osteopenia in comparison with DM without osteopenia.     

Alkaline phosphatase isoenzymes of liver and bone origin are incompletely resolved by wheat-germ-lectin affinity chromatography

We used wheat-germ-lectin affinity chromatography as a tool to investigate the structure of alkaline phosphatase (ALP, EC 3.1.3.1) and to obtain fractions enriched in either bone or liver ALP activity. Liver and bone isoenzymes in serum samples were incompletely resolved except that the activity in the nonretained fraction (fraction 1) always represented pure liver isoenzyme and constituted a larger percentage of total activity in pooled sera with increased liver ALP activity than in pooled sera with increased bone activity. In contrast, a more avidly retained ALP activity, presumably with high glycosylation, was found in human serum with high activity of bone ALP. Using a solid-phase immunoassay, we examined the fractions obtained from the wheat-germ-lectin-Sepharose 4B column to determine whether the isoenzyme preference of the monoclonal antibody was markedly influenced by the degree of glycosylation. Whether samples contained high proportions of liver or of bone isoenzyme activity, the nonretained fraction contained a higher percentage of liver ALP, whereas the more strongly bound fraction contained a higher percentage of bone ALP. Except for eluted fractions that either contained no detectable N-acetylglucosamine or the highest percentage of it, the avidity of the liver-isoenzyme-specific monoclonal antibody for ALP seemed to be independent of the degree of glycosylation, suggesting that the epitope for monoclonal antibody may be expressed in some structure other than the carbohydrate moieties.     

Age and sex distribution of alkaline phosphatase isoenzymes by agarose electrophoresis

We separated isoenzymes of alkaline phosphatase (ALP; EC 3.1.3.1) in 1383 sera of normal individuals (ages 4-65 years) by agarose electrophoresis with the Isopal system (Analis). As expected, the predominant isoenzyme in children was of bone origin, and almost all (99%) of the children had low activities of a second bone fraction, "bone variant" ALP. The "bone variant" disappeared after age 17 in girls and after age 20 in boys. The highest (median) bone ALP activity was reached at age 9 to 10 in girls and at age 13 to 14 in boys, followed by a gradual decline in girls and a steep decline in boys. During adulthood, activity of the bone fraction was constant and no significant differences were observed between sexes, neither for bone nor for liver ALP activity. The latter remained almost unchanged throughout life. We observed no high-Mr ALP activity in children, whereas sera from 60% of the adults contained low activities of high-Mr ALP. Intestinal ALP (soluble form) and "intestinal variant" ALP (hydrophobic form) were frequently present, in 21% and 37% of all samples, respectively. No significant differences were observed between age groups and sexes for the intestinal isoenzymes.     

血清肝型碱性磷酸酶糖链分析方法的改进及其临床应用

本文建立了直接用唾液酸酶切除血清碱性磷酸酶（ＡＬＰ）糖链末端的唾液酸后，用蔓陀萝凝集纱（ＤＳＡ）一支亲和层析柱分析其糖链结构的简单方法，用该法分析了１０例正常人和４９例不同肝病患者血清肝型碱性磷酸酶糖链结构，发现只有肝癌血清ＡＬＰ含有与ＤＳＡ强结合的“多天线（ｍｕｌｔｉａｎｔｅｎｎａｒｙ）结构，这一结果与以臆繁琐的序列凝集亲和层析法所得的结果相同，本文不仅再次证实了ＤＳＡ分析血清ＡＬＰ糖链结构在区