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61.
Summary Techniques and protocols are described for the generation of genetically modified cells that can be used for gene therapy. Primary fibroblast cultures are established from skin biopsies, maintained in culture, frozen for long-term storage, and retrieved when necessary. Retroviral packaging cell lines are generated by transfection of DNA into retroviral packaging cells by calcium-phosphate precipitation method or by lipofection method. To generate cell lines expressing high titer virus, individual colonies of cells are cloned and the virus titer is determined. Virus collected from packaging cells expressing high titer virus is then used to infect primary fibroblasts. To obtain fibroblast cell lines expressing high amounts of transgenes, individual cells can be cloned to generate clonal cell lines. Although the methods described here are for fibroblasts, the same methods or modification of the methods can be used for other cell types. 相似文献
62.
The intracerebroventricular administration of the 17 amino acid peptide nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the NOP receptor (previously referred to as ORL-1 or OP4 receptor), reduces voluntary 10% ethanol intake in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats. Studies aimed at the pharmacological characterization of the receptor, which mediates the effect, have shown that the C-terminal 13 amino acid sequence is crucial for activity and that the selective NOP receptor antagonist [Nphe(1)]N/OFQ(1-13)NH(2) blocks the effect of N/OFQ on ethanol drinking. In place conditioning studies, N/OFQ abolishes the conditioned place preference (CPP) induced by ethanol in msP rats, or by morphine in nonselected Wistar rats; these findings suggest that N/OFQ is able to abolish the rewarding properties of ethanol and morphine. Moreover, N/OFQ inhibits reinstatement of alcohol-seeking behavior induced to electric footshock stress, as well as reinstatement of alcohol-seeking behavior induced by ethanol-paired cues. Together, these findings suggest that N/OFQ and its receptor may represent an interesting target for pharmacological treatment of alcohol abuse. 相似文献
63.
Alcohol dehydrogenase II (ADH II, structural genealcB) was purified from a strain H1035,biA1; alcE1; alc500 alcD1, which produces 100-times more ADH II activity than thealcAalcR deletion strain (alc500). Antibodies were raised against this ADH, and were used to screen a cDNA library in gt11. We have isolated the gene for an ADH which is over-expressed in H1035, and which we believe to be thealcB gene; cDNA and genomic clones were sequenced. The sequence contains three introns and encodes a protein of 367 amino acids. This protein shows a clear level of identity to a range of alcohol dehydrogenases, but is no more closely related to the ADH I and ADH III previously described inA. nidulans than to the ADHs ofS. pombe andS. cerevisiae. The significance of consensus sequences found in the 5 region of the gene is discussed in relation to the regulation of the gene. 相似文献
64.
Juhani Juntunen Heikki Teräväinen Kalervo Eriksson Andreo Larsen Matti Hillbom 《Virchows Archiv : an international journal of pathology》1979,383(3):241-252
Summary The effects of variable dietary thiamine concentrations (deficient, normal, surplus) on the development of alcoholic neuromyopathy in rats exposed for 36 weeks to 10–25% (v/v) ethanol or water (control group) as the sole drinking fluid were studied by histological and electrophysiological methods.Abnormalities in the structure of the sciatic nerve (phagocytosis, myelin abnormalities, increase in nonspecific cholinesterase activity) and tibial muscles (angular atrophic fibers, group atrophy, fibre necrosis) developed more frequently in animals on diets deficient in thiamine than in animals on diets with normal or surplus thiamine, and more frequently in animals drinking alcohol and water than in those drinking water alone. No differences were observed between the different groups in the number of perivascular sympathetic nerves, in the motor nerve conduction velocities and in the muscle fibrillation potentials.Thus, thiamine deficiency, established as a significant reduction of red blood cell transketolase activity, seems to have a deleterious effect on the peripheral nerves and muscles. The effect is enhanced by the simultaneous consumption of ethyl alcohol. 相似文献
65.
帕罗西汀合并认知疗法治疗酒依赖伴发抑郁及对戒酒的影响 总被引:1,自引:0,他引:1
目的评价帕罗西汀合并认知疗法治疗酒依赖伴发抑郁的疗效及对戒酒的影响。方法将98例酒依赖伴发抑郁的患者随机分为研究组和对照组,分别用认知疗法合并帕罗西汀、单用帕罗西汀治疗4个月。用汉密顿抑郁量表(HAMD)评定疗效;用复饮率评定戒酒效果。结果治疗1个月后,研究组HAMD评分16.79±6.50与对照组相近17.88±6.59(P>0.05)。治疗4个月后HAMD评分10.76±5.32、明显低于对照组14.54±5.12(P<0.01);临床疗效(痊愈率44.44%、显效率22.22%、有效率33.34%)明显高于对照组(21.74%、21.74%、56.52%)(P<0.05);复饮率(24.44%)低于对照对照组(45.56%)(P<0.05)。结论认知疗法合并帕罗西汀治疗酒依赖伴发抑郁疗效较好,戒酒效果较好。 相似文献
66.
丹参提取物F对大鼠乙醇性胃粘膜损伤的影响及其机制的研究 总被引:4,自引:1,他引:4
在大鼠乙醇性胃粘膜损伤模型上,丹参提取物F 0.1ml/200g/min iv5分钟可明显降低胃粘膜伊文斯兰含量(P<0.01)和被单星蓝染色的面积(P<0.01)及胃粘膜损伤程度(P<0.01)。用TS-200组织光谱分析仪测定大鼠胃粘膜表层血液量和Hb氧饱和度,表明丹参提取物F可维持大鼠胃粘膜在60%乙醇(容积比)后血液量和Hb氧饱和度基本不变。结果提示:丹参提取物F有抗大鼠乙醇性胃粘膜损伤作 相似文献
67.
Prof. H. Hofmann Dr. C. Bothge Dr. S. Haselwarter Prof. L. Heinemann Dr. W. Barth Dr. E. Schüler Prof. U. Laaser Dr. M. Siegel Prof. F. C. Luft Die Intersalt-Forschungsgruppe aus BRD und DDR 《Journal of molecular medicine (Berlin, Germany)》1990,68(13):655-663
Summary The relationships between body mass index (BMI) and age, alcohol consumption, 24-hr urinary electrolyte excretion, and BP were studied in 588 subjects from three German centers participating inIntersalt, a highly standardized, previously reported protocol. Men and women aged 20–59 were sampled in Bernried, FRG; Cottbus, GDR; and Heidelberg, FRG. The subjects from the three centers did not differ in BMI, level of education, physical activity, cigarette- or alcohol-consumption patterns, or urinary Cl excretion. Mean Na excretion was 167, 147, and 172 mmol/24 hr in Bernried, Cottbus, and Heidelberg, while mean K excretion was 72, 55, and 73 mmol/24 hr, respectively. The excretion of these electrolytes was significantly lower in Cottbus than in Bernried or Heidelberg. BMI increased progressively in men with age; in women BMI plateaued until the 5th decade, after which it increased to equal that of men. In individual centers, the excretion of electrolytes was correlated with BML Sodium and chloride excretion were highly correlated. The data from each individual center were fitted to a multiple regression model. Age, BMI, sex, and alcohol consumption entered the model. 相似文献
68.
Gerald E. McClearn 《Behavior genetics》1993,23(2):223-230
Under a variety of rubrics (e.g., complexity, self-constructing systems, dissipative structures), interest has recently burgeoned in applying principles of complex systems to a wide variety of scientific issues. A major concern is with emergent properties of systems not derivable from the properties of components of the systems. In this paper, some elementary aspects of systems considerations are applied to phenomena of alcohol pharmacogenetics. It is likely that whole new families of informative phenotypes can be generated by this approach. 相似文献
69.
饮酒对小鼠睾丸生精小管一氧化氮合酶、增殖细胞核抗原表达及细胞凋亡影响的研究 总被引:3,自引:0,他引:3
目的 探讨酒精对小鼠睾丸的组织结构、内皮型一氧化氮合酶 (eNOS)、增殖细胞核抗原 (PCNA)及细胞凋亡的影响。 方法 用 5 %、10 %及 15 % 3种不同浓度的酒精作用于 2 2d龄小鼠 ,取睾丸做石蜡切片、HE染色 ;用免疫组织化学方法检测睾丸eNOS、细胞增殖的变化 ;TUNEL法检测细胞凋亡的变化 ,并进行统计学分析。 结果 随着酒精浓度的增大 ,睾丸组织结构发生明显改变 ,生精小管的直径逐渐减小 ,eNOS阳性细胞面积密度逐渐增大 ,单位面积内PCNA阳性细胞和凋亡细胞数目增加 ,高浓度酒精组与其他组差异极显著 (P <0 0 1)。 结论 酒精可使生精小管的直径变小 ,eNOS及PCNA表达增强 ,凋亡细胞增加并随酒精浓度的增大而变化加重。这可能是过量饮酒导致生精细胞减少 ,生殖能力降低的重要因素。 相似文献
70.
F. Sztark P. Latry A. Quinton C. Balabaud P. Bioulac-Sage 《Virchows Archiv : an international journal of pathology》1986,409(3):385-393
Summary Alcohol induces morphological changes in the endothelial and perisinusoidal cells at the fibrotic stage of alcoholic liver diseases. Directly or indirectly, through hemodynamic disturbances linked to the enlargement of steatotic hepatocytes, alcohol may modify this barrier before the onset of fibrosis. Liver biopsies were obtained from control and from alcoholic patients and perfusion-fixed. Volume and surface densities of endothelial cells, perisinusoidal cells and their processes were measured. Liver histology was normal in the 2 groups except for steatosis in the alcoholics. Volume densities represented 8.2%, 4.7% and 3.2% of the sinusoid in controls for endothelial cells, perisinusoidal cells and their processes whereas surface densities represented respectively 0.5, 0.23, 0.21 m2/cm3 of sinusoid. Morphometric values were not significantly different in the alcoholic patients. In none of the alcoholic patients did fine morphological studies of sinusoidal cells give any indication of the possible evolution of the alcoholic disease towards fibrosis. These results indicate that in the group of patients studied, alcohol, before the fibrotic stage, did not significantly alter the sinusoidal barrier. 相似文献