Rapid and simple isolation of DNA from agarose gels

Summary We present a simple method for the isolation of DNA from agarose gels that is economic, fast, and independent of electrical equipment. DNA fragments of up to 6 kb can be easily extracted within 5 min using a disposable plastic syringe and filter paper. Total extraction of DNA fragments between 10 and 20 kb in size is achieved by concentrating the DNA flushed from the gel in a DNA-binding column.     

大鼠腹裂模型肠管损伤的研究

目的研究先天性腹裂的肠管受损害情况，探讨该病术后并发症的原因。方法利用大鼠腹裂模型，运用组织学、生化学和免疫组织化学方法，分析腹裂胎鼠肠管的组织结构，DNA和蛋白质，细胞增生和凋亡等方面的改变。结果共获得腹裂胎鼠16只，对照胎鼠21只。与对照组相比，腹裂鼠肠管变短、充血水肿、粘连，肠壁表面纤维覆盖，壁内胶原沉积，DNA总量下降，蛋白质总量基本不变，细胞增生率下降，凋亡率上升。结论腹裂的肠管损伤是多方面的，是术后肠管运动和吸收功能异常的原因，大鼠的腹裂模型是对先天性腹裂的病因、病理等方面研究的合适工具。     

1-Methylnicotinamide stimulates cell growth and inhibits hemoglobin synthesis in differentiating murine erythroleukemia cells

Exposure of murine erythroleukemia cells (MELCs) to nicotinamide (NA) or its synthetic analog N′-methylnicotinamide (N′-MN) reduces cell growth and induces terminal differentiation, marked by increased heme and globin accumulation. On the contrary, 1-methylnicotinamide (1-MN), the primary metabolite of excess NA, was found to stimulate cell growth and reduce spontaneous differentiation of cultured MELCs. Log phase MELCs exhibited up to 50% higher cell density above untreated cells when cultured for up to 96 h with 2.5 mM 1-MN. When combined with NA or several chemically-unrelated inducers of hemoglobin synthesis in cultured MELCs, 1-MN reduced the globin mRNA levels and heme accumulation by 40–80%. 1-MN was able to inhibit heme production if present during only the first 24–48 h after NA exposure. Pre-treatment with 1-MN could not confer resistance of cells to effects of NA, suggesting the inhibition is reversible. Commitment to differentiate in semisolid medium by the most potent inducer, 5 mM N′-MN, was inhibited up to 95% by 2.5 mM concentrations of 1-MN. It appears that 1-MN has opposing effects on growth and induction of differentiation than those seen in MELC cultures exposed to NA or N′-MN.     

精子洗涤对精液中HBV DNA含量的影响

目的：检查HBV感染者精液是否存在乙型肝炎病毒（HBV），探讨精子洗涤是否能去除其中的HBV DNA。方法：用实时荧光定量PCR（FQ-PCR）法检测57名血清HBsAg阳性患者精液中HBV DNA，采用Percoll非连续梯度离心及上游法分离其中21份精液，并对洗涤后标本进行HBV DNA检测。结果：62份精液标本中有15份HBV DNA阳性，阳性率为24.19%;21份用于洗涤的精液标本中有，有7份HBV DNA阳性，洗涤后仍有6份标本阳性。结论：部分HBV感染者精液具有传染性，精子洗涤不能完全去除精液中的乙肝病毒。将HBV感染者的精液用于人工授精或体外受精应慎重。     

生物医学测量及控制技术新进展

介绍生物医学测量及控制技术领域中的一些研究进展。     

巢湖水有机提取物致鱼红细胞DNA损伤的初步观察

目的　研究巢湖水有机提取物致突变性。方法　单细胞凝胶电泳技术测定鱼红细胞DNA损伤效应。结果　巢湖源水引起慧星细胞的百分率最高 (5 7.2 5 % ) ,滤前水最低 (2 7.6 3% ) ,出厂水经过二次加氯后慧星细胞的百分率有所上升 (4 4 .0 0 % )。结论　巢湖源水具有潜在致突变性 ,经混凝、活性炭吸附及沉淀处理后其DNA损伤作用有所下降 ,但氯化消毒可增加水中有机提取物的DNA损伤作用。     

Brain and kidney of progressive multifocal leukoencephalopathy patients contain identical rearrangements of the JC virus promoter/enhancer

The kidneys of six progressive multifocal leukoencephalopathy (PML) patients were examined by PCR amplification for the presence of JC virus. Amplification of three different areas of the viral genome from multiple samples of each kidney revealed three that were positive for the virus. The use of a PCR-based typing assay on all tissue samples, and cloned sequences from the viral coding region from each positive kidney showed that the same viral genome was present in the kidney as in the brain of the patient. Regulatory region clones all had the archetypal promoter/enhancer structure. However, when PCR fragments from the regulatory region were digested with a restriction enzyme which cuts in region D, the region most often deleted in PML-type promoters, a low level of undigested DNA remained. This DNA refractory to digestion had a rearranged sequence identical to that of the unique rearranged promoter in the brain of each patient. © 1994 Wiley-Liss, Inc.     

Cerivastatin inhibits fetal calf serunvinduced DNA synthesis in cultured rat mesangial cells

SUMMARY: Inhibition of mevalonate synthesis by several statins has been shown to suppress DNA synthesis in glomerular mesangial cells. In the present study, we investigated the effect of a new statin, cerivastatin, on fetal calf serum (FCS)-induced DNA synthesis of cultured rat mesangial cells. Cultured rat mesangial cells were stimulated by 10% FCS in the presence or absence of cerivastatin and mevalonate. 5-bromo-2-deoxyuridine (BrdU) incorporation was used to assess DNA synthesis. the present study showed that 10% FCS caused marked stimulation of DNA synthesis in the mesangial cells. Cerivastatin inhibited FCS-stimulated BrdU incorporation in a dose-dependent manner. IC₅₀ was approximately 1 umol/L. Exogenous mevalonate, farnesyl pyrophosphate and geranylgeranyl pyrophosphate significantly prevented the inhibitory effect of cerivastatin on DNA replication. It appears that cerivastatin, by inhibiting the synthesis of mevalonate, may suppress DNA synthesis in the mesangial cells.     

Andrology: Application of different in-situ hybridization detection methods for human sperm analysis

The detection of some types of aneuploidy in human spermatozoacan be based on the use of the fluorescence in-situ hybridizationtechnique (FISH). One of the crucial steps for FISH is to achievea proper decondensation and denaturation of the DNA in the specimen,so as to obtain efficient hybridization results. However, afterDNA decondensation the morphology of sperm heads is partly distortedand the majority of the tails is lost. This situation leadsto problems in the distinction between disomic and diploid spermatozoa,as well as between abnormal spermatozoa and somatic cells. Double-and triple-target FISH can partly solve this discriminationproblem. To improve these procedures we adapted the steps ofdecondensation and visualization of the single sperm cells.Firstly, DNA decondensation with 25 mM dithiothreitol in 1 MTris at pH 9.5 resulted in sperm cells with intact morphologyof both the head and the tail, and allowed efficient single-,double- and triple-target ISH to be performed. Secondly, weapplied a novel detection method, based on enzyme immunocyto-chemicalreactions, with coloured precipitation products. Thirdly, thisISH procedure was combined with Diff-Quik staining and bright-fieldmicroscopy. This absorption method has the advantage of a permanentsignal, and the adapted cytoplasmic staining of the sperm plasmamembrane allows the visualization of the outline of the singlespermatozoon. Using this approach, therefore, it is possibleto discriminate between disomic, diploid and abnormal spermatozoa,somatic cells and spermatozoa that overlap, because the morphologyof the cells is not distorted and the tails of the spermatozoaare intact and properly visualized.     

Higher-order organization and compartmentalization of satellite DNA PIM357 in species of the coleopteran genus Pimelia

The PIM357 satellite DNA family is present in 26 Pimelia taxa (Tenebrionidae, Coleoptera) with endemic congeneric species from the Canary Islands showing higher interrepeat variability than continental ones. In this paper, we compare the repetitive DNA sequences of a Canarian species that has distinct subfamilies of repeat units, P. radula ascendens, with another without such subfamilies, P. sparsa sparsa. The chromosomal localization of the repeat units and the comparison of the variability of randomly cloned monomers to the one estimated by comparing repeat units from dimers and trimers suggest the absence of satellite subfamilies in P. sparsa sparsa. Hence, the repeat units of this species seem to be uniformly and randomly distributed throughout all chromosomes out of one chromosomal pair. On the contrary, P. radula ascendens shows four divergent subfamilies of repeat units supported by several diagnostic nucleotide substitutions. These subfamilies seem to form four distinct repeat units: monomer subfamily 1, monomer subfamily 4 and two higher-order units (dimer linking subfamily 1 and 4, and dimer linking subfamily 2 and 3). Moreover, monomers of subfamily 1 are present in three chromosomal pairs only. We discuss the effect of different potential factors acting in the concerted evolution and the genomic organization of stDNA sequences in these taxa. This revised version was published online in July 2006 with corrections to the Cover Date.