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111.
The importance of oxygen in maintaining the functional integrity of hepatocytes has been well established in a variety of experimental models, such as in vivo , perfused liver and isolated hepatocytes. However, one of the shortcomings of these systems is their short life span. Therefore, we have examined the effects of long-term hypoxia on cellular adenine nucleotide content and cellular functions, such as albumin production, urea production and DNA synthesis, in adult rat hepatocytes in primary culture. Hepatocytes were cultured at a density of 11 × 104 and 5 × 104 cells/0.18 mL per cm2 for the study of albumin and urea production and DNA synthesis, respectively, at various oxygen tensions (20, 12, 8 and 5%) for 24 h. Cellular ATP content in cultured hepatocytes in hypoxia gradually declined, corresponding to the decrease in oxygen tension, and the cellular ATP level at 5% oxygen was approximately 20% of that at 20% oxygen. Albumin production also decreased in parallel with the decrease in cellular ATP content in cultured hepatocytes in hypoxia. However, even when cellular ATP content gradually declined corresponding with the decrease in oxygen tension in cultured hepatocytes in hypoxia, such as at 8 or 5% oxygen, urea production remained at a high level; in contrast, DNA synthesis was completely suppressed. These results suggest that the cellular ATP content decreases in cultured hepatocytes during long-term hypoxia in relation to oxygen tension and that the relationship between decreased ATP levels and liver function in cultured hepatocytes during hypoxia differs for albumin production, urea production and DNA synthesis.  相似文献   
112.
113.
应用不同浓度人白细胞干扰素处理10例慢性乙型肝炎病人外周血淋巴细胞,观察病人姐妹染色单体互换(SCE)频率的动力学变化.结果表明,人白细胞干扰素处理组SCE平均频率明显低于对照组,差异高度显著(P<0.01);慢性迁延性肝炎和慢性活动性肝炎处理组之间SCE平均频率无显著性差异;但人白细胞干扰素处理低浓度组SCE平均频率低于高浓度组,差异不显著(P>0.05).提示人白细胞干扰素具有诱导HBV损伤人体DNA的修复作用;大剂量人白细胞干扰素对HBV损伤人体DNA的修复未必有益.  相似文献   
114.
目的探讨维吾尔族和汉族弱精子症病人精子mtDNA~(4977bp)缺失的相关性。方法本文采用聚合酶链反应(PCR)技术对60例标本分两组,维、汉各30例,其中精子活力正常标本(对照组)和弱精子症患者(观察组)各15例进行了精子线粒体DNA(mtDNA)缺失的研究。结果发现15例做内对照的维族精子活力正常人中仅有3例(20%)有mtDNA~(4977bp)缺失;而15例维吾尔族弱精子症患者中有10例(67%) mtDNA~(4977bp)缺失;两组4977bp缺失频率比较有显著性差异(P<0.05)。15例做内对照的汉族精子活力正常标本中无mtDNA~(4977bp)缺失;15例汉族弱精子症患者中有12例(80%)mtDNA~(4977bp)缺失。两组4977bp缺失频率比较差异有显著性(P<0.05)。同时发现维吾尔族男性不育症与汉族男性不育症mtDNA~(4977bp)缺失频率无显著性差异。结论维吾尔族和汉族mtDNA~(4977bp)缺失在弱精子症的发病中可能起重要作用。  相似文献   
115.
慢性HBV感染患者精子中HBVDNA存在研究的初步报告   总被引:1,自引:0,他引:1  
用Southern Blot方法,我们研究了13份慢性HBV感染患者(8份慢迁旰,5份慢活肝)和2份对照精液标本中HBV DNA存在状况。提取物一半用限制酶HindⅢ消化,各行琼脂电泳,转移到纤维膜上,然后用~(32)P—HBV—DNA探针分子杂交。在4份精子內检出HBV DNA,3份为游离形式,1份为整合形式。结果表明HBV DNA可以存在精于内,游离状态HBV DNA提示有传染性、整合序列HBV DNA的存在提示有通过生殖细胞遗传的可能。  相似文献   
116.
本文用显微分光光度计对39例手术切除的不同组织学类型的人体肺癌标本癌细胞核的DNA含量进行了测量。其中高、中、低分化的鳞癌分别为3、6和5例;高、中、低分化的腺癌分别为3、5和5例;未分化癌大细胞型和小细胞型各为3例和6例。结果发现各类型肺癌的DNA含量均有统计学上的差异(P<0.01)。各DNA含量直方图的峰值部位及分布范围亦不同。从而提示了根据DNA含量的不同,可能有助于肺癌组织学类型及其分化程度的判别:而DNA含量直方图对肺癌类型的判别亦可能是有益的。  相似文献   
117.
Evidence from both experimental carcinogenesis and studies in human cirrhotic liver suggest that defective repair of the promutagenic DNA base lesion, O 6-methylguanine, is a factor in the multistep process of hepatocellular carcinogenesis. Ubiquitous environmental alkylating agents such as N-nitroso compounds can produce O 6-methylguanine in cellular DNA. Unrepaired, O 6-methylguanine can lead to the formation of G ? A transition mutations, a known mechanism of human oncogene activation and tumour suppressor gene inactivation. Combined treatment of rodents with an agent producing O 6-methylguanine in DNA, and an agent promoting cell proliferation, leads to development of hepatic nodules and hepatocellular carcinoma (HCC), cell division, hence DNA replication, being required for the propagation of tumorigenic mutation(s) in hepatocyte DNA. The paramount importance of O 6-methylguanine in hepatocellular carcinogenesis is indicated by the observation that transgenic mice engineered to have increased hepatic levels of repair enzyme O 6-methylguanine-DNA methyltransferase (MGMT) are significantly less prone to hepatocellular carcinogenesis following alkylating agent treatment. Cirrhosis is a universal risk factor for development of human HCC, and a condition that is characterized by increased hepatocyte proliferation as a result of tissue regeneration. Levels of the human repairing enzyme for O 6-methylguanine were found to be significantly lower in cirrhotic liver than in normal tissue. In accord with findings from animal models, this suggested a mechanism in which persistence of O 6-methylguanine due to defective DNA repair by MGMT, together with increased hepatocyte proliferation, might lead to specific gene mutation(s) and hepatocellular carcinogenesis. Screening for the presence and persistence of O 6-methylguanine in human DNA presently involves formidable technical difficulty. Indications are that such limitations might be overcome by the use of an ultrasensitive method such as immuno-polymerase chain reaction (PCR). This approach should allow parallel measurement of DNA adduct and repair enzyme in routine liver biopsy samples. It might also enable investigation of O 6-methylguanine in human genes specifically associated with hepatocellular carcinogenesis. Given the wide variation in human MGMT levels observed between individuals, tissues, and cells, this technology should be adapted to permit the ultrasensitive localisation and measurement of adducts and repairing enzyme in liver biopsy tissue sections. Ability to ultrasensitively measure O 6-methylguanine, and its repair enzyme, should prove valuable in the risk assessment of cirrhotic patients for developing hepatocellular carcinoma. Received for publication on July 6, 1998; accepted on Aug. 12, 1998  相似文献   
118.
Human lymphocytes (HL) as well as lymphocytes (RL), hepatocytes (RH), and gastric mucosa cells (GM) of Sprague-Dawley rats were treated in vitro for 1 h with methylmercury chloride (MMC, 0.5–4 μg/ml) and dimethylmercury (DMM, 5–40 μg/ml). The cytotoxicity of the two organic mercury compounds was assessed by dye exclusion, and the extent of induced DNA fragmentation was measured with a single-cell microgel electrophoresis assay. Both MMC and DMM induced DNA damage and cytotoxicity in a dose-related manner in HL, RL, and GM. MMC was more effective in causing a significant increase in median DNA migration than DMM at doses yielding approximately the same degree of cytotoxicity. In rat hepatocytes the MMC-induced DNA damage was, however, lower than in the other cells. An analysis of repair kinetics following exposure to 2 μg/ml MMC was carried out in human lymphocytes obtained from an adult male donor. The bulk of DNA repair occurred 90 min after in vitro exposure, and it was about complete by 120 min following cessation of exposure. Finally, in order to have a basis for extrapolating to the human situation, in vivo studies were performed with Sprague-Dawley rats, also assessing the DNA damage and cytotoxicity in the lymphocytes and gastric mucosa cells. These in vivo results after oral exposure may be directly compared to the in vitro data obtained in the same cells. © 1993 Wiley-Liss, Inc.  相似文献   
119.
Renal gene transfer techniques are being developed as a novelexperimental approach to understand the pathogenesis of renaldisease and to potentially develop new therapeutic tools. Wereview the currently available technology to introduce foreigngenetic material into renal tissue, i.e., retroviral, adenoviral,and liposomal transfer systems with their respective advantagesand caveats. Today, the transfer efficiency of these methodsappears to be sufficiently high to study the effects of transducedgenes on renal function and morphology in rat kidney. This willallow (i) the elucidation of the function of genes on the courseof renal disease in experimental animal models and (ii) themodulation of local expression of endogenous genes which presumptivelycontribute to renal pathology in these models. One strategyto accomplish this aim is the use of recombinant DNA technologyto design antisense DNA constructs or oligonucleotides, whichinterfere with the renal expression of target genes. We willalso discuss some of the shortcomings of the currently usedtechniques with respect to potential therapeutic use of genetransfer systems and gene modulation.  相似文献   
120.
Mitochondria were isolated from the dimorphic zygomycete Mucor racemosus by differential centrifugation. DNA from the organelles was purified by cesium chloride-ethidium bromide isopycnic centrifugation. Examination of the mitochondrial DNA by electron microscopy revealed a circular chromosome approximately 63.8 kbp in circumference. The chromosome was digested with restriction endonucleases and the resulting DNA fragments were separated by agarose-gel electrophoresis. Electrophoretic mobilities and stoichiometry of the fragments indicated a mixed population of mtDNA molecules each with a size of about 63.4 kbp. Physical maps were constructed from analyses of fragments generated in single and double restriction digests and from the hybridization of fragments to probes for the large and small mitochondrial rRNA genes from Saccharomyces cerevisiae. The Mucor mitochondrial chromosome was found to exist in the form of two flip-flop isomers with inverted repeat sequences encoding both rRNA genes.  相似文献   
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