Sexual dimorphism in rat cerebrum and cerebellum: different patterns of catalytically active creatine kinase isoenzymes during postnatal development and aging

During postnatal development, maturation and aging the Wistar rat cerebrum and cerebellum synthesize, in a different sex-dependent manner, catalytically active dimeric cytosolic (c) muscle-type (MM) and heart-type (MB) creatine kinase (CK), besides the supposedly sole type brain-specific (BB) CK. In both sexes, typical and atypical neuromuscular cCK isoenzymes were present during the study for 26 months. As in rat heart, females showed more cerebral cCK variants (41%) in comparison to males. Female rats exhibited about 93% more cerebellar variants of cCK isoenzymes as compared to males. The male cerebellum showed predominantly BB- and MB-CK during the whole study in comparison to the female one that contained all neuromuscular cCK variants. Only female rats showed decreases and increases of cerebral CK specific activity. In contrast to males, coinciding with the weaning period, cerebral female CK activity decreased 45% from 14 to 21 days and increased about 3-fold in female rats and only 1.3-fold in males from 21 to 45 days of age. Contrary to the remarkable 4-fold increase of chicken brain CK specific activity exhibited at old age, the rat did not show another cerebral CK activity increase during senescence in either sex. However, sex differences of CK specific activity appeared in the cerebellum at all ages. From the sex-specific plateau phase at 45–60 days until 2.2 years of age, about a 41% independent increase of cerebellar CK specific activity was observed in both sexes. After puberty, the differential cerebellum–cerebrum values of CK specific activity were higher for female rats than males during youth, adulthood and senescence. The present work shows that in rat cerebrum and cerebellum, production of ATP through anaerobic transphosphorylation by the CK/PC system is sex-and age-specific, especially in the cerebellum, when glycolysis and the Krebs cycle lose capacity. As in rat heart, under physiological conditions at all ages the several cCK isoenzymes do participate in a gender-specific manner, in favor of females, in diverse functions of the different cell compartments of glial and neuronal cells with regard to their high and fluctuating energy demands not completely covered by anaerobic and aerobic glycolysis.     

Isoelectric focusing on cellulose acetate membrane: a separation procedure for alkaline phosphatase isoenzymes

An isoelectric focusing technique for the separation of alkaline phosphatase isoenzymes on cellulose acetate membrane is described. Optimal conditions for isoelectric focusing were established by changing ampholine concentration and focusing conditions. Bone, liver, intestinal, and placental isoenzymes can be resolved into vacious sub-bands in a pH range of 4.1 to 5.2. These sub-bands were correlated with the findings of electrophoretic isoenzyme separation. The whole procedure proves very simple to perform and comparatively time saving (4 h). This procedure may help clarify the problems of ALP isoenzyme differention when electrophoretic patterns are unresolved.     

常用急性心肌梗死诊断标志物临床应用价值比较

①目的　比较几种常用的急性心肌梗死 (AMI)诊断标志物的临床应用价值。②方法　检测 46例AMI病人及 62例心绞痛 (AP)病人血清中的心肌肌钙蛋白Ⅰ (cTnⅠ )、肌红蛋白 (Mb)、肌酸激酶 (CK)、肌酸激酶同工酶 (CK MB)含量 ,并进行对比分析。③结果　AMI组cTnⅠ、Mb、CK、CK MB各指标阳性率分别为 93 .48%、97.83 %、65 .2 2 %、76 .0 9% ;AP组各指标阳性率分别为 1 .68%、48.39%、32 .2 6 %、1 4 .52 %。Mb与cTnⅠ对AMI诊断灵敏度差异无显著性 (χ2 =0 .345 ,P >0 .0 5) ;两者高于CK、CK MB对AMI诊断灵敏度 (χ2 =4.1 2 8～ 9.552 ,P <0 .0 1、0 .0 5)。cTnⅠ与CK MB对AMI诊断特异度分别为 90 .32 %、85 .48% ,两者比较差异无显著意义 (χ2 =0 .683 ,P >0 .0 5) ;两者高于Mb与CK对AMI诊断特异度 (χ2 =9.538～ 2 2 .545 ,P <0 .0 1 )。④结论　cTnⅠ是诊断AMI较好指标     

安氟醚对体外循环下心内直视手术心肌缺血再灌注损伤的影响

目的：研究安氟醚对体外循环心内直视手术期间心肌缺血再灌注损伤的影响。方法：选择择期行心瓣膜置换的风湿性心脏病患者30例，随机分为3组，Ⅰ组、Ⅱ组在主动脉阻断前分别吸入1．0MAC、0．5MAC的安氟醚，Ⅲ组为对照组静脉给芬尼太。在围术期不同时点作血肌酸磷酸激酶(CK—MB)、脂质过氧化产物(MDA)、超氧化物歧化酶(SOD)测定。结果：3组患者CK—MB、MDA、SOD值均于主动脉开放即刻升高，与麻醉前、块血前比较差异有显著性(P均＜0．05)，于主动脉开放后10、30min达最大值，并高于主动脉开放即刻(P＜0．05)，术后24h虽低于主动脉开放10min时(P＜0．05)，但仍高于麻醉前。在主动脉开放即刻、10min，Ⅰ组患者CK—MB值显著高于Ⅱ组和Ⅲ组(分别P＝0．0220、0．0108、0、0202和0．0295)；主动脉开放60min时，Ⅰ组患者CK—MB值也高于Ⅲ组(P＜0．0001)；在主动脉开放10、30min时，Ⅰ组MDA值明显高于Ⅱ组(分别P＝0．0060、0．0364)；在主动脉开放10min时，Ⅰ组SOD值也高于Ⅱ、Ⅲ组(分别P＝0．000l和0．0084)。各组CK—MB值与主动脉阻断时间至正相关(r＝0．55—0．8l，P均＜0．05)；CK—MB值与SOD、MDA值之间无明显相关性；MDA与SOD间Ⅰ组和Ⅱ组均呈正相关(r＝0．8l、0．7l，P均＜0．01)。结论：体外循环下心内直视手术期间，存在心肌缺血再灌注损伤，并伴有氧自由基产生增加和机体抗氧化的储备提高。大剂量安氟醚吸入可能对心肌酶谱及氧自由基的产生有一定的影响。     

酶组化显示睾丸LDH－X方法的再探讨

对小鼠睾丸冰冻切片ＬＤＨ－Ｘ的显示方法作了进一步探讨。实验表明，以Ｌ－２－羟基－３－甲基戊酸（ＨＭＶ）为特异性底物，能相当有效地显示睾丸ＬＤＨ－Ｘ的分布，且该底物的配制方法简便易行。在显示ＬＤＨ－Ｘ的孵育液中采用萘酚蓝（ＮＢ）作中间递电子体较吩嗪甲基硫酸酯更具优越性。     

血清电解质、CKBB及血乳酸水平与新生儿缺氧缺血性脑病严重程度的相关性分析

分析血清电解质、肌酸激酶同工酶(CKBB)及血乳酸水平与新生儿缺氧缺血性脑病严重程度的相关性。80例缺氧缺血性脑病患儿中轻度患儿27例、中度患儿26例、重度患儿27例。以本院同期健康的足月新生儿60例为对照组。结果显示与对照组新生儿比较,观察组新生儿氯离子及钙离子水平明显降低(P<0.05),CKBB和血乳酸水平明显升高(P<0.05);重度患儿的氯离子及钙离子水平明显低于中度患儿和轻度患儿(P<0.05),CKBB和血乳酸水平明显高于中度患儿和轻度患儿(P<0.05);血清中氯离子和钙离子与缺氧缺血性脑病严重程度呈现负相关性(P<0.05),但CKBB和血乳酸与缺氧缺血性脑病严重程度呈明显的正相关性(P<0.05)。结果说明血清氯离子和钙离子、CKBB及血乳酸水平与新生儿缺氧缺血性脑病严重程度呈明显的相关性。     

脑型肌酸激酶同工酶测定在非心源性疾病的应用

目的 探讨脑型肌酸激酶同工酶(CK-BB)在肿瘤、自身免疫性疾病的筛查以及器官损伤定位中的意义. 方法 免疫抑制法检测血清标本心肌型肌酸激酶同工酶(CK-MB)、肌酸激酶(CK)活性,计算二者比值.随机选取CK-MB/CK＞50%的非心源性疾病的血清标本90份及30份对照血清,用CK同工酶琼脂糖凝胶电泳进行分离、鉴定. 结果 血清CK-MB假性增高的标本多见于肿瘤、自身免疫病,经电泳分离及可见光扫描后,肿瘤和自身免疫性疾病组的CK-BB含量明显高于对照组,差别具有统计学意义(P＜0.01),血清CK-BB增高情况与其在组织器官中的分布相符. 结论 血清CK-BB的检测在肿瘤、自身免疫性疾病的辅助诊断以及器官损伤定位中具有重要意义.     

不同宿主来源的华支睾吸虫同工酶比较观察

目的:通过对不同宿主华支睾吸虫的同工酶研究,了解其在不同宿主中的变化情况.方法:应用浓度梯度聚丙烯酰胺凝胶板状电泳(CG-PAGE)对50 d虫体的乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、酯酶(EST)、过氧化物酶(PO)进行电泳观察,通过凝胶成像软件对酶(蛋白)带的迁移率、相对浓度分布、光密度等结果进行分析.结果:来自不同宿主(大鼠、犬、豚鼠、兔、猫)的虫体主要酶带特征基本相同,但次要酶带存在一定差异.结论:不同宿主的华支睾吸虫同工酶存在着一定的变化,实验结果为华支睾吸虫在不同宿主中的发育、生理、生化提供实验资料.     

心肌肌钙蛋白Ⅰ的临床应用研究

探讨心肌肌钙蛋白Ⅰ在缺血性心脏病中的诊断与鉴别方面的临床应用价值。结果表明：心肌肌钙蛋白Ⅰ对急性心肌梗死的诊断灵敏度高于肌酸激酶、肌酸激酶同工酶ＭＢ，乳酸脱氢酶、谷草转氨酶及α－羟丁酸脱氢酶；其特异性在稳定型心绞痛组与肌酸激酶和肌酸激酶同工酶ＭＢ相当，而较α－羟丁酸脱氢酶、乳酸脱氢酶和谷草转氨酶为高。此外，心肌肌钙蛋白Ⅰ对不稳定型心绞痛也有一定的诊断价值。提示心肌肌钙蛋白Ⅰ是一种心肌损伤的特异性标志物