进化水平不同的动物和人血清乳酸脱氢酶同工酶电泳图谱的比较研究

应用聚丙烯酸胺垂直板梯度凝胶电泳分析了进化水平不同的17种动物和人血清乳酸脱氢酶(LDH)同工酶.17种动物是鲤鱼、中华大蟾蜍、蝮蛇、罗斯蛋鸡、山鸡、紫貂、银狐、貉、梅花鹿、马鹿、黑白花奶牛、新疆细毛绵羊、猪、马、大耳白兔、昆明小鼠和猴.在同一凝胶板、同一条件下,对这些动物和人血清LDH同工酶谱分析表明,LDH同工酶谱在各类动物之间存在明显差异.进化比较低级的动物(杂交鲤鱼除外),血清LDH同工酶谱带迁移率较小,接近阴极;反之,进化比较高级的动物,同工酶谱带迁移率较大,接近阳极.亲缘关系相近的动物,LDH同工酶谱相似.表明LDH同工酶谱带的差异与动物间的亲缘关系有关.同种不同个体的动物LDH同工酶谱差异不明显,只有杂交鲤鱼个体LDH同工酶谱存在明显差异.     

初生北京鸭4种组织LDH同工酶的分析

本文用薄层聚丙烯酰胺凝胶等电聚焦电泳对初生北京鸭的４种组织进行同工酶分析，结果表明初生北京鸭肝脏具有１１条区带，胸肌具有９条区带，肾脏具有１０条区带，心脏具有９条区带。从它们的Ａ亚基和Ｂ亚基的相对百分量的比值中可看出心脏和肾脏中的Ｂ亚基含量比肝脏中的多，相反，肝脏的Ａ亚基含量比心脏和肾脏含量多。胸肌的Ａ亚基和Ｂ基含量则是相近似的。     

Ontogenetic Development of Creatine Phosphokinase in Skeletal Muscles and Heart from Pigs

Creatine Phosphokinase (CPK) in striated muscles shows only small changes in activity before birth. After birth and during the first month of extrauterine life the activity increases rapidly. The largest increase is seen in muscles with a glycolytic energy metabolism (m. long, dorsi) and the smallest in muscles with an oxydative energy metabolism (m. flexor dig. ped. sup.). The differences between these groups of muscles are statistically significant. In heart tissue the increase in CPK activity is lower, the levels amounting to 40 to 47 % of those in striated muscles.Early in fetal life only the BB isoenzyme is found in striated muscles. Synthesis of M subunits of GPK starts between day 76 and 65 before birth and increases rapidly after this time leading to disappearance of the BB isoenzyme 24 days prior to birth and of the MB isoenzyme at birth. In muscles with an oxydative as well as in muscles with a glycolytic metabolism all GPK activity after birth is caused by the MM isoenzyme.All three isoenzymes are present in heart tissue at the earliest prenatal stage investigated, the pattern being dominated by the BB isoenzyme. During further differentiation the MM isoenzyme increases and the BB isoenzyme decreases. The development is completed during the first month after birth with a final isoenzyme composition of 81 % MM and 19 % MB isoenzyme. kw|Keywords|k]pigs; k]ontogenesis; k]creatine phosphokinase; k]activity; k]isoenzymes     

罗斯及印第安河鸡发育时期组织LDH同工酶变化

罗斯及印第安河鸡在胚胎第3d的混合胚中出现LDH_(1～3)3种同工酶(3条区带,梯度凝胶电泳),或LDH_(1～4)4种同工酶(6～7条区带,等电聚焦电泳);第7d检出心肌、胸肌和脑的5种同工酶(LDH_(1～5),5～7条区带);第12d检出肝脏的LDH_(1～5)5种同工酶(8或15条区带).组织LDH同工酶区带数随发育过程明显增多,同种鸡同一组织不同发育时期的LDH同工酶活性变化相差显著或极显著.在相同发育时期,印第安河鸡胸肌和肝脏的LDH_1相对活性均明显高于罗斯鸡(P<0.01).两种鸡心肌、胸肌、肝和脑组织LDH同工酶的等电点主要分布在pH6.0～8.2内.     

三种药物对猪棘头虫LDH同工酶的抑制试验

根据猪棘头虫与宿主（猪）血清的ＬＤＨ同工酶的差异，用０．１％硝硫氰醚、０．１％丙硫咪唑和０．１％吡喹酮分别对猪棘头虫雄虫、雌虫和猪血清的ＬＤＨ同工酶进行了抑制试验，结果除硝硫氰醚对蛭形巨吻棘头虫雄虫、雌虫的ＬＤＨ同工酶有抑制作用外，其余的试验组与对照组均无差异。硝硫氰醚对雄虫ＬＤＨ同工酶的第２、３、４带和雌虫ＬＤＨ同工酶的第２、３带有不同程度的抑制作用，特别是雌虫ＬＤＨ同工酶的第２带最敏感，在０．１％的浓度下，该酶带被完全抑制，即没显示出来，硝硫氰醚对雄虫和雌虫ＬＤＨ同工酶的极弱带（它们各自的第１带）均无抑制作用；临床实验也显示出硝硫氰醚对治疗猪棘头虫病有一定疗效，这些表明硝硫氰醚抑制蛭形巨吻棘头虫的ＬＤＨ同工酶活力可能是该药物对该虫体的作用机理之一。     

绵羊组织器官LDH同工酶特异性研究

本研究利用不连续聚丙烯酰胺凝胶电泳法对绵羊的８种组织器官ＬＤＨ同工酶的分布及相对活性进行了分析测定。比较ＬＤＨ同工酶的酶谱特征。结果表明，绵羊的ＬＤＨ同工酶分布特征不同于其它动物，其相对活性有显著差异。骨骼肌中ＬＤＨ５活性比ＬＤＨ１强，Ｍ亚基比例大于Ｈ亚基；肝脏和心脏中ＬＤＨ１的活性最强。     

Vergleich der histologischen Struktur der Kompakta der langen Röhren-knochen bei Maus, Hamster, Ratte, Meerschweinchen, Kaninchen, Katze und Hund während der Altersentwicklung

This paper reports on histological examinations of the diaphyseal structure of the long bones (humerus, radius, ulna, os femoris and tibia) of mouse, hamster, rat, guinea pig, rabbit, cat, and dog using six animals in three different age groups. There are considerable differences in structure between the species. The results show that with increasing height in the evolution of the species and period of life, as well as advancing age, the differentiation and complexity of bone structures increases. The differences in structure between the localization of bones within the individual species are not considerable. The species comparison results in great similarities between the bone structures of each of mouse and hamster, of rat and guinea pig, as well as of cat and dog. The bone structure of the animals examined becomes more similar to the human structure from mouse, hamster, rat, guinea pig, rabbit, and cat to dog.     

Measurement of certain plasma and serum enzymes in the horse

The study of the use of serum or plasma enzymes in the horse is reviewed.

Analyses of such enzymes in the horse have been performed mainly for the detection and monitoring of abnormalities which affect liver tissue and muscle fibres. Examples of muscle disorders are post-anaesthetic forelimb lameness and the clinical myopathy syndrome which varies from a mild “tying-up” to a severe paralytic myoglobinuria (EPM). Measurements have also been made during training periods in an endeavour to assess the effects of exercise on muscle tissue. One of the benefits of such a study could be the prediction of the period of optimum performance.

The two enzymes most frequently used for. muscle studies have been glutamic oxaloacetic transaminase (GOT) (aspartate aminotransferase) and creatine Phosphokinase (CPK). The former, while not muscle specific and less sensitive than CPK to small amounts of muscle tissue damage, has a slower clearance rate from blood than CPK. Determination of both enzymes together has some advantages in determining the prognosis in cases with severe muscle damage. Later studies have included aldolase, which is a sensitive skeletal muscle specific enzyme and lactic dehydrogenase (LDH), which is widely distributed by various tissues at high concentrations. Most of the work on serum LDH has centred on its separation into 5 isoenzymes, and on relating the isoenzyme pattern to the various tissues. More studies on this, and on the isoenzymes of CPK, are required.

Apart from cost, one of the major problems in enzymology has been lack of a standardisation of laboratory methods which has led to a wide range of published normal values. Reliance on such values may lead to difficulties in interpretation, particularly when serum changes are small.     

Lactate dehydrogenase and its isoenzymes in the tissues and sera of clinically normal dogs

The total activity of lactate dehydrogenase (LDH) and the percentage distribution of its isoenzymes in the tissues and sera of clinically normal adult dogs are presented. Total LDH activity was greatest in skeletal muscle followed by heart muscle, kidney, small intestinal mucosa, liver, lung, pancreas and bone. Each tissue had a unique isoenzyme pattern and the proportions of the isoenzymes in serum suggested that liver is the source of normal serum LDH. The tissue isoenzyme patterns were similar to those obtained by other authors in human beings, horses, cattle, sheep and cats although in liver, differences between ruminants and monogastric animals including dogs were evident. The data presented provide a basis for the interpretation of serum LDH isoenzyme patterns in canine disease.     

Activities of enzymes in the malate-aspartate shuttle and the isoenzyme pattern of lactate dehydrogenase in plasma and peripheral leukocytes of lactating Holstein cows and riding horses

The activities of the enzymes involved in the malate-aspartate shuttle and lactate dehydrogenase (LDH) and the pattern of the isoenzymes of LDH were determined in plasma and peripheral leukocytes of lactating Holstein cows and thoroughbred riding horses as representative herbivorous animals. In the horse plasma, LDH activities were significantly lower and AST activities were significantly higher than those in the cow plasma. The specific activities of cytosolic malate dehydrogenase (MDH), LDH and AST in the horse leukocytes were higher than those in the cows. The cytosolic ratio of MDH/LDH activity (ML ratio) in the horse leukocytes was significantly lower than that in the cow leukocytes owing to significantly higher activities of LDH. The ML ratio was considered to reflect the difference in energy metabolism in leukocytes between cows and horses. The plasma LDH isoenzyme patterns of cow and horse showed the characteristic as herbivorous animals with dominance of LDH-1, -2 and -3. The LDH isoenzyme patterns with dominance of LDH-3 and -4 in the horse leukocytes were remarkably different from those in the cow leukocytes. There were significant differences in activities of malate-aspartate shuttle enzymes, ML ratio and LDH isoenzyme patterns in the cytosolic fractions of leukocytes between the lactating cows and the riding horses.     

马关节滑液、血清乳酸脱氢酶及其同工酶和它们在疾病状态下的变化

用king氏法和聚丙烯酰胺凝胶电泳法测得马关节滑液和血清乳酸脱氢酶(LDH)及其同工酶正常值。滑液和血清5种LDH同工酶电泳迁移率相同。各种LDH同工酶的分子量和热稳定性均有明显差异。抗尿素作用,血清和滑液LDH同工酶基本相似。Mg~(++)是LDH激活剂,最佳激活浓度为10~(-3)M;Zn~(++)、Ca~(++)、Cu~(++)、Hg~(++)、谷胱甘肽、半胱氨酸、碘乙酸、EDTA等为LDH抑制剂。在四肢的急性或亚急性关节和骨疾病时,滑液LDH明显升高,其同工酶谱随疾病发生部位不同而不同,并与血清一致,但慢性关节疾病时LDH及其同工酶变化则不一致。     

LDH and CK isoenzyme patterns in the blood plasma of horses with elevated CK, LDH and AST activities

The distribution of LDH and CK isoenzymes in blood plasma of ten clinically sound Thoroughbreds with reasonable performance and without elevated clinico-chemical blood variables (reference group) was compared with 57 Thoroughbreds, which had histories of mild locomotor disturbances and/or poor performance and had elevated CK, LDH and/or AST activities (trial group). The trial group was subdivided according to the number of altered blood variables and in the groups with two as well as three altered blood variables also according to the extent of alteration of the total CK activity. The pattern of LDH and CK isoenzyme distribution in the blood plasma of the reference group was the following: 22% LDH1, 36% LDH2, 34% LDH3, 6% LDH4 and 2% LDH5 as well as 75% CK1 and 15% CK2. The remaining 10% of the plasma electropherogram could not be alloted to any one of the two CK bands. All trial groups built showed a similar pattern of changes in their isoenzyme distribution independent on kind and combination of altered enzyme activities. The shares of CK1, LDH4 and LDH5 were significantly increased whilst the shares of CK2, LDH1 and LDH2 decreased. A multiple analysis of variance demonstrated that only increased total CK activities had a pronounced effect on distribution of LDH and CK isoenzyme patterns in the trial group (p less than 0.01 for LDH2, LDH3, LDH4, CK1 and p less than 0.05 for CK2). The conclusion of the study was that the altered distribution pattern of LDH and CK isoenzymes of the trial group signalized an increased skeletal muscle membrane leakage.     

豆状囊尾蚴与细颈囊尾蚴和宿主（兔）的同工酶,蛋白质比较研究

用聚丙烯酰胺凝胶圆盘电泳分离出豆状囊尾蚴囊液乳酸脱氢酶同工酶4～5条酶带,酯酶同工酶1条酶带,蛋白质6～9条区带;豆状囊尾蚴囊壁和头节乳酸脱氢酶同工酶3～4条酶带,酯酶同工酶无,蛋白质17条区带;细颈囊尾蚴囊液乳酸脱氢酶同工酶1～4条酶带,酯酶同工酶1～3条酶带,蛋白质7～10条区带;宿主——兔腹水乳酸脱氢酶同工酶5条酶带,酯酶同工酶3～5条酶带,蛋白质13～16条区带。并作了豆状囊尾蚴与细颈囊尾蚴囊液同工酶、蛋白质和豆状囊尾蚴与宿主的同工酶、蛋白质相互关系的详细分析,结果表明豆状囊尾蚴与细颈囊尾蚴的同工酶和蛋白质具有差异和两者可能存在交叉免疫;豆状囊尾蚴的酶蛋白和蛋白质来源独立于宿主,但在一定程度上受宿主的影响。     

Growth and Development of Testes in Domesticated and Hybrid(wild × domesticated)Yak Bulls

Testicular tissue was studied in domesticated yak and in wild yak × domesticated yak(F1 and F2)bulls at 6,12,18 and 24 months of age by stereology for quautitative histology and by comparative studies on the lactate dehydrogenase(LDH)isoenzyme spectrum, LDH activity and the percentage content of LDH isoenzymes in testes. The results indicated that all three types of yak were similar in both the characteristics of ultrastructure of testicular tissue and the degree of germ cell development. The process of spermatogenesis was initiated and sperm were produced for the first time at the age of 12months in all three yak types. All yak types exhibited similar age-related, increases in weight of testes,volume density of both the seminiferous tubules and seminiferous epithelium and height of seminiferous epithelium but all these indexes were slightly affected by season. The number of LDH isoenzyme bands after electrophoresis varied with age as follows: four bands were present at 6 months of age, five bands at 12 and 18 months and six bands at 24 months of age. The sixth band was LDH -x ,which appeared between the LDH4 and LDH5 band. These results indicated that yak bulls reach sexual maturity at 24months of age and that puberty is not affected by infusion of wild strains.     

Lactate dehydrogenase isoenzyme patterns in bovine liver tissue

Liver lactate dehydrogenase (LDH) isoenzymes in normal and diseased cows were analyzed electrophoretically. This method (histoelectrophoresis) was improved for the direct analysis of tissue LDH isoenzymes. The mean values of LDH 1 through LDH 5 in the livers of normal cows were 31.7, 24.8, 27.3, 12.8, and 3.3%, respectively. In cases with hydropic degeneration of the liver, the patterns revealed increases of LDH 1 and LDH 2 as compared to normal cows. The patterns showed a decrease of LDH 1 and an increase of LDH 2 in fatty change of the liver. Congestion of the liver alone decreased LDH 1 and increased LDH 3, LDH 4 and LDH 5. Necrosis of the liver decreased LDH 1 and LDH 2, and increased LDH 3, LDH 4 and LDH 5. It was suggested that the functional hepatocellular damage due to anoxia might be a important factor of the change of liver LDH isoenzyme patterns. We have attempted to standardize the LDH isoenzyme patterns by using a computer under various conditions. In cases with hydropic lesions, the diagnostic sensitivity (DS) was 53% (28 of 53 cases) and the predictive value of positive tests (PVPT) was 100% (28 cases of 28 cases selected). In cases with congestive or necrotic lesions, DS was 41% (9 of 22 cases) and PVPT was 69% (9 cases of 13 cases selected).     

LDH and LDH Isoenzymes of Synovial Fluid in the Horse

LDH is an intracellular enzyme, which when cells degenerate is released to the extracellular spaces and body fluids. Cells and organs in the mammalian body differ from each other with respect to their LDH isoenzyme patterns. These circumstances have led to the use of LDH isoenzyme determinations in laboratory diagnostic work. In the present investigation total LDH activity and LDH isoenzyme distribution in equine synovial fluid from healthy joints, joints with serous arthritis, osteochondrosis dissecans and arthrosis, were determined. The fluids from the diseased joints differed from normal synovial fluid with respect to total LDH activity, and the different joint diseases each seemed to give rise to a characteristic isoenzyme pattern. In order to examine possible sources of the increased LDH activity and altered isoenzyme patterns, blood plasma, red and white blood cells, synovial membrane and articular cartilage were also studied. It was found that LDH4 and LDH5 were present in high amounts in articular cartilage, and an increase in these isoenzymes was the most characteristic feature in synovial fluid from joints with arthrosis. The results were discussed in view of possible diagnostic value of isoenzyme determinations on synovial fluid.     

Different patterns in the histology and autofluorescence of the Harderian glands of the Syrian Hamster, rat, mouse, Mongolian gerbil and guinea pig

The purpose of this study was to compare the natural fluorescence in the Harderian glands of the Syrian hamster, rat, mouse, Mongolian gerbil and guinea pig (both sexes). For each species, 10 animals (five males and five females) were used. Histological autofluorescence studies were performed using a fluorescence microscope (450-490 nm filter). Two different types of fluorescent cells were observed in both hamster (type AFI high intensity and type AFII, low fluorescence) and rat (type AFI, low fluorescence and type AFII, high fluorescence) Harderian glands. The fluorescence was basally located in all mice cells, whereas it was observed near the epithelial cell nuclei in the Mongolian gerbil (occupying two-thirds and one-third of the cells in males and females, respectively). A high intensity of fluorescence was present throughout the acinar cells in the guinea pig. The patterns of fluorescence identified exhibited a sexual dimorphism in all species studied. These results demonstrate that the Harderian glands of the animal species examined exhibit a variety of histological autofluorescence patterns.     

Serum total amylase activity and its isoenzyme fractions in newborn pigs with relation to growth and iron dextran-treatment.

Serum amylase activity was analyzed chromogenically (blue starch method) and its isoenzyme was separated by cellulose acetate membrane electrophoresis in newborn pigs comparing with those in adult pigs. Serum amylase activity in newborn pigs was about one half of adults' and increased with age. Serum amylase isoenzymes were separated into 4 fractions from the cathode. The isoenzymes were also classified in 5 types on the basis of the combination of these 4 fractions. Incidences of the fractions and the types were not significantly different between adults and newborns. After the iron dextran treatment, serum amylase activity as well as levels of erythrocytes, hemoglobin and packed cell volume increased. The rates of increase in hemoglobin and packed cell volume were more significant in the double treatment group than in the single treatment group, while serum amylase activity increased similarly in these 2 groups. The increase in serum amylase activity seemed to indicate that the development of amylase-producing organs was reinforced by the iron compound treatment. It is thought that serum amylase activity may be an indicator of healthy growth in newborn pigs.     

Lactate dehydrogenase isoenzymes in the lungs of sheep with acute and chronic pneumonia

Total lactate dehydrogenase and the absolute and percentage levels of its isoenzymes were measured in lung lesions and macroscopically normal areas of lung from lambs with chronic proliferative exudative pneumonia and acute pasteurella pneumonia. Lung lesions had a higher total enzyme activity which was associated mainly with increases in the activity of the LDH4 and LDH5 isoenzymes, particularly in chronic pneumonia, and gave lung lesions a considerable potential for altering the serum isoenzyme distribution. Thus, the nature of any changes in the serum isoenzyme distribution will depend on whether the isoenzymes are released from abnormal or normal areas of lung. This appears to be the first report on lactate dehydrogenase isoenzymes in ovine pneumonia.     

Alkaline phosphatase and its isoenzymes in the tissues and sera of normal dogs

The total alkaline phosphatase (AP) activity and the pattern of its isoenzymes were studied in the tissues and sera of normal adult dogs. Small intestine mucosa showed the greatest total AP activity followed by kidney, bone, pancreas, liver, lung, skeletal muscle and heart muscle. After separation by agarose gel electrophoresis, each tissue showed only one isoenzyme except lung which showed two. The tissue isoenzymes, in decreasing order of migration distance towards the anode, were as follows: fast lung isoenzyme, liver or slow lung isoenzyme, the group consisting of skeletal muscle, bone, small intestine and pancreas isoenzymes and, finally, the kidney isoenzyme. Two isoenzymes occurred in serum. The major band corresponded to liver and the slow lung isoenzyme, while the minor band was considered to be the corticosteroid-induced isoenzyme, previously thought to be absent from normal serum.The AP isoenzyme patterns in lung and skeletal muscle and the presence of an isoenzyme migrating an identical distance to the corticosteroid-induced isoenzyme do not appear to have been reported before in normal dogs.