青霉素G酰化酶α亚基Ser177的突变对酶活性的影响

用盒式突变和定点突变对大肠杆菌青霉素G酰化酶α亚基177位ser进行了突变研究,结果发现所挑选的突变体均无酶的活力,这一结果可能可以用来解释Ser 177附近肽段和一些青霉素结合蛋白青霉素结合区在一级结构上保持同源性的原因。     

The inhibition of catalase by glutathione

Reduced glutathione (GSH) inhibited catalase activity in a dose-dependent manner. DL-dithiothreitol (DL-DTT) and dithioerythritol (DTE) also inhibited catalase activity. The inhibition of catalase by GSH and DL-DTT could be reduced by NADPH. Polyacrilamide gel electrophoresis demonstrated the inhibition was partially reversible. The inhibition of catalase by GSH appeared to be partly due to superoxide radicals, since it was inhibited by active manganese superoxide dismutase, but not by heat-inactivated enzyme. Other chemical species also appear to take part in the inhibition, but they could not be identified.     

Detection of IgG and IgM antibodies with ELISA technique in human trichomoniasis

The direct wet mount examination of vaginal secretion, widely applied for the diagnosis of Trichomonas vaginalis infection in woman patients, is rapid and economical, however, the sensitivity of this technique is not so high. In this study enzyme-linked immunosorbent assay (ELISA) was employed for the detection of serum anti-T. vaginalis IgG and IgM antibodies from 30 vaginal trichomoniasis patients and 30 non-infected healthy persons. The results were as follows: 1. Serum ELISA-IgG value was 0.37 +/- 0.134 (Mean +/- S.D.) in vaginal trichomoniasis patients and 0.21 +/- 0.054 in healthy controls (p less than 0.005), and the sensitivity and specificity of ELISA for serum IgG antibody were 70.0% and 96.7%, respectively. 2. Serum ELISA-IgM value was 0.33 +/- 0.177 (Mean +/- S.D.) in vaginal trichomoniasis patients and 0.11 +/- 0.051 in healthy controls (p less than 0.005), and the sensitivity and specificity of ELISA for serum IgM antibody were 70.0% and 96.7%, respectively. 3. The ELISA-IgG values showed a significant correlation with ELISA-IgM values (r = 0.77, p less than 0.005). With above results, it is assumed that ELISA is a reliable method for the diagnosis of T. vaginalis infection and simultaneous measurement of serum IgG and IgM with this technique is recommended.     

长江中游鱼类寄生棘头虫区系的研究

经过3年10次调查,剖检湖北省宜都、黄冈两处江段所产72种鱼类,共计766尾。收集棘头虫10种,其中包括2新种和1新组合,即蛇鮈新棘吻虫(新种)Neoechinorhynchus saurogobi sp.nov.,长江丽棘虫(新种)Brentisentis yangtzensis sp.nov.(Illiosontidae),鲤丽棘虫(新组合)B.cyprini comb.nov.。对长江中游鱼类寄生棘头虫区系的特点进行了分析和探讨。     

Loss of tumorigenic potential by human lung tumor cells in the presence of antisense RNA specific to the ectopically synthesized alpha subunit of human chorionic gonadotropin

A clonal strain of human lung tumor cells in culture (ChaGo), derived from a bronchogenic carcinoma, synthesizes and secretes large amounts of alpha (alpha) and a comparatively lower level of beta (beta) subunit of the glycoprotein hormone, human chorionic gonadotropin (HCG). ChaGo cells lost their characteristic anchorage-independent growth phenotype in the presence of anti-alpha-HCG antibody. The effect of the antibody was partially reversed by addition of alpha-HCG to the culture medium. ChaGo cells were transfected with an expression vector (pRSV-anti-alpha-HCG), that directs synthesis of RNA complementary to alpha-HCG mRNA. The transfectants produced alpha-HCG antisense RNA which was associated with the reduced level of alpha-HCG. Transfectants also displayed several altered phenotypic properties, including altered morphology, less mitosis, reduced growth rate, loss of anchorage-independent growth, and loss of tumorigenicity in nude mice. Treatment of transfectants with 8,bromo-cAMP resulted in increased accumulation of alpha-HCG mRNA, no change in the level of alpha-HCG antisense RNA, release of the inhibition of [3H]thymidine incorporation, and restoration of anchorage-independent growth phenotype. The overexpression of c-myc, observed in ChaGo cells, was unaffected by the reduced level of alpha-HCG. These results suggest that ectopic synthesis of the alpha subunit of HCG plays a functional role in the transformation of these human lung cells.     

睾丸间质细胞—研究自体吞噬的一种正常细胞模型

In the present study, we tried to estimate, in a semiquantitative way, the relative frequency of the autophagic activity in various cell types under physiological condition. The results indicated that the highest activity appeared to be in the Leydig cells of rat testes. Autophagosomes were frequently observed in electron microscope photographs of Leydig cells, which provide a good model to study the autophagocytosis in normal cells. The autophagic process in Leydig cells was observed with the electron microscope in preparations treated to show CMPase activity. The mode of formation of autophagosomes in Leydig cells can be divided into three steps. Step 1, flattened membranous elements expand to enclose a small cytoplasmic territory to form pre-autophagosome. Step 2, The double membrane profile of the pre-autophagosome then completely encloses the cytoplasmic territory to form early autophagosome in which structurally normal organelles are contained. Step 3, the transformation of an early autophagosome into a late one is accompanied by the loss of one of the two delimiting membranes, the partial disintegration of the enclosed content and simultaneous acquisition of acid phosphatase activity. The enzymatic reactivity is acquired following a close association with the lysosomes. The late autophagosome then reaches the cell surface and appear to exocytose their residual content.     

A procedure for the construction of linear restriction fragment maps of a 50- to 100-kb DNA.

A simple and effective procedure for the construction of linear restriction fragment maps was developed. Using a two-enzyme digestion, two-dimensional (2-D) electrophoresis procedure, all the restriction fragments in a 50- to 100-kb DNA can be individually resolved and displayed on a 2-D plane. This 2-D gel pattern, with appropriate markers, provides a fixed set of x, y coordinates for each fragment obtained from the single and double digestion as well as the relationship between the two steps. A matrix is constructed from the 2-D pattern. The vertical column shows all the singly digested individual fragments and their sizes obtained from each restriction enzyme treatment, and the dividing horizontal row shows all the doubly digested DNA fragments and their sizes after treatment with two enzymes. The order of arrangement is always from the smallest to the largest fragments. Using this matrix, two linear DNA restriction maps for these two enzymes can be simultaneously constructed in a self-reconfirming manner. As examples for this procedure, we describe the construction of two linear restriction fragment maps, a combination of EcoRI and BamHI digestion as well as a combination of EcoRI and HindIII digestion of lambda-phage DNA.     

粉纹夜蛾核型多角体病毒诱导的胸苷激酶特性研究

粉纹夜蛾(Trichoplusia ni)核型多角体病毒(TnNPV)感染草地贪夜蛾(Spodop-tera frugiperda)细胞后能诱导提高胸苷激酶的活性。不论是正常或感染细胞中的胸苷激酶都可将脱氧胞苷磷酸化,酶活最适pH及Mg~( )离子浓度值基本相同,DEAE-纤维素和Cibacron blue-sepharose柱层析所得酶活性图谱亦相似。TnNPV在胸苷激酶缺陷型的草地贪夜蛾细胞中能正常复制,但被感染细胞不具有胸苷激酶活性。由此可以确定,经TnNPV感染诱导后,活性有所提高的胸苷激酶不是病毒基因编码的。