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。     

海南文昌汉族人群中两种葡萄糖-6-磷酸脱氢酶基因突变的研究

目的:分析海南文昌人群中葡萄糖-6-磷酸脱氢酶基因1376G→T、95A→G突变。方法:应用硝基四氮唑蓝定量法进行G6PD缺乏症的筛查,用等位基因特异PCR检测1376G→T、95A→G突变。结果:在358位海南文昌汉族人中,发现G6PD缺乏症患者20例,其中9例患者有1376G→T突变,3例患者有95A→G突变。结论:1376G→T、95A→G突变是文昌人群中常见的突变。     

REGULATING FUNCTION OF COENZYMIZATION AND DECOENZYMIZATION OF THE LACTATE DEHYDROGENASE ISOZYMES IN THE MOUSE TISSUES DURING HYPOXIA

Objective. To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.Methods. Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by the specific staining technique. LDH (1 -5) relative percentage enzymatic activity (RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner.Results. The RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymes LDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH,(H4), LDH2(H3M) increased, while RPEA of LDH5(M4) in various tissues decreased prominently except the cardiac muscle, and that of LDH4(HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypoxia treated cardiac muscle specimen was made, activity subbands originated regularly in the isozyme patterns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands), LDH4 (1-3 subbands), LDH5 (2-4 subbands).     

Regulation of neuronal differentiation in neuron-enriched primary cultures from embryonic rat cerebra by platelet activating factor and the structurally related glycerol ether lipid, dodecylglycerol.

Primary cultures enriched in neurons dissociated from embryonic rat cerebra were used to demonstrate that platelet activating factor and the structurally related ether glycerolipid, dodecylglycerol, are readily taken up in small amounts by neurons and that they stimulate the differentiation of neurons. The stimulation of neuronal differentiation was observed as a precocious development of axon-like extensions which correlated with a concentration-dependent increase in neuronal-specific enzyme activities. This stimulation of morphological and neurochemical factors by either platelet activating factor or dodecylglycerol was almost completely abolished by triazolam, a known inhibitor of platelet activating factor function. Neither platelet activating factor nor dodecylglycerol at the concentrations used to achieve stimulation of neuronal differentiation compromised the plasma membrane, as indicated by the lack of leakage of cytoplasmic lactic acid dehydrogenase.     

Multidimensional analysis of metabolism contributions involved in running track tests

It is difficult to interpret the training induced changes in middle-distance running, since numerous aerobic and anaerobic determinants of the performance are interdependent. Several aerobic and anaerobic tests are available but their results, particularly those from anaerobic tests, may be discordant, not providing univocal interpretation of training. The purpose of this study is to use a multidimensional approach to distinguish aerobic and anaerobic capacities assessed by two running tests on a track: the maximal anaerobic running test (MART) and V(O2max) tests. Eleven runners carried out two maximal tests on a synthetic track before and after a 4-week training period: (i) a maximal test to determine V(O2max), the velocity associated with V(O2max) (vV(O2max)) and the velocity at the lactate threshold (v(LT)), (ii) a maximal anaerobic running test to estimate anaerobic capacity. An all-out test run at v(LT)+50% of the difference between v(LT) and vV(O2max), known to be affected by both aerobic and anaerobic energy production, was used to test this approach. A principal components analysis (PCA) shows that two components (i.e., aerobic and anaerobic) explained 79% of the variation in the physiological variables. The PCA suggests that V(O2max) and MART tests assess the aerobic and the anaerobic capacities, respectively. In contrast, the performance in the all-out test is affected by both aerobic and anaerobic energy production. The PCA shows that v(LT) and DeltaP (difference between the maximal power of the MART and V(O2max)) are clear markers of the long-term endurance and the anaerobic capacity, respectively. This multidimensional approach can be a useful way to disentangle the aerobic and anaerobic components of track tests.     

Testicular seminoma with human chorionic gonadotropin production

Testicular seminoma with elevated serum human chorionic gonadotropin level (hCG-positive seminoma) is regarded as more malignant than marker-negative seminoma, although Its prognosis is still unclear. To clarify the malignant potential of seminoma with hCG production, the serum levels of the beta subunit of hCG (β-hCG) and lactic acid dehydrogenase (LDH) were examined in 35 and 40 patients, respectively, and the Immunohistochemical expression of β-hCG examined in 45 tumors. The elevation of the LDH serum level correlated to the Invasive status, metestatic status and poor outcome, while that of the serum β-hCG level correlated only to the metastatic status. Immunohistochemical expression of β-hCG was observed in syncytiotrophoblastic giant cells in 11 tumors and a few mononuclear seminoma cells In 36 tumors. Expression was not associated with the malignancy potential, except where the expression In mononuclear cells Inversely correlated to the invasive status. These results suggest that most seminomas produce a slight amount of hCG; that an elevated hCG serum level Indicates the pressnce of metastatic tumors and mainly reflects an increase in tumor volume but not in cellular malignancy potential; and that the LDH serum level, rather than hCG, is more useful as a prognostic indicator for patients with seminoma.     

谷氨酸脱氢酶,胆碱氧化酶和葡萄糖-6-磷酸酶对溶酶体中~(67)Ga积累的影响(英文)

目的:比较正常肝组织与肝癌AH 109A,吉田肉瘤中谷氨酸脱氢酶,胆碱氧化酶和葡萄糖-6-磷酸酶的活力对~(67)Ga摄取与积累的影响;方法:制备~(67)Ga枸橼酸溶液给大鼠静注后处死大鼠,制备亚细胞悬液,液闪计数器测定放射活度.结果:~(67)Ga的放射活性在正常肝组织溶酶体中(55％积聚)显著高于肝癌AH109A(32％积聚)和吉田肉瘤(18％)积聚.谷氨酸脱氢酶的活力在正常肝组织,肝癌和吉田肉瘤分别是1830±s 320 U·L~(-1),23±s 6 U·L~(-1)和7±s 2 U·L~(-1);胆碱氧化酶的活力分别是46±s 10 U·L~(-1),25.0±s 0.4 U·L~(-1),2.0±0.4 U·L~(-1);葡萄糖-6-磷酸酶活力分别是2550±s 180 U·L~(-1),84±s 14 U·L~(-1),78±s13 U·L~(-1).结论:正常肝组织中溶酶体酶活力很强,对~(67)Ga的积累起较大作用.癌变组织酶活力降低而作用减弱.吉田肉瘤细胞无肝细胞特点,其溶酶体对~(67)Ga积累作用不大.     

乳酸脱氢酶实验检测细胞新陈代谢的实验研究

目的 探索LDH实验检测细胞活力的可行性。方法 原代培养骨髓细胞和软骨细胞，用LDH实验测定上述两组细胞的活力，并与镜下活体观察到细胞的生长状况相比较。与目前比较成熟的测定细胞活力的MTS实验的测得的值相比较。结果 LDH实验对上述两组细胞的活力的测定结果与镜下活体观察到的结果相符合。与MTS实验的测得的结果经统计学处理无显著差异。结论 LDH实验可用于细胞活力的直接测定，而对活细胞的生存、繁殖无影响。     

邻苯二甲酸二乙基己酯对新生小鼠睾丸及Leydig细胞影响的实验研究

目的:探讨邻苯二甲酸二乙基己酯(DEHP)对新生小鼠睾丸及Leyd ig细胞形态结构及功能的影响。方法:DEHP分别以低、中、高3组剂量[100、200、500 mg/(kg.d)]灌胃作用于怀孕12 d到产后3 d(GD12~PND3)的KM母鼠,观察DEHP对新生雄性仔鼠体重、睾丸重量、Leyd ig细胞形态结构和3β-羟基类固醇脱氢酶(3-βHSD)活性、酶反应面积的影响。结果:DEHP作用于母鼠后,其雄性子代幼鼠体重和睾丸重量减轻,睾丸Leyd ig细胞形态、超微结构发生改变;高剂量组Leyd ig细胞数量明显增多;低、中剂量组睾酮合成关键酶3β-HSD酶活性下降,酶反应面积减小,但高剂量组在仔鼠出生后15 d时酶活性降低[(吸光度值(0.154±0.011)vs空白对照组(0.222±0.013),P<0.01],而酶反应面积增大[(6 303.0±745.6)μm2vs空白对照组(5 091.4±214.4)μm2,P<0.01)]。结论:DEHP能影响新生雄性小鼠体重、睾丸重量、Leyd ig细胞的形态结构和3β-HSD活性,具有抗雄激素效应。