Purification and Properties of Component B of 2,4,5-Trichlorophenoxyacetate Oxygenase from Pseudomonas cepacia AC1100

Pseudomonas cepacia AC1100 degrades 2,4,5-trichlorophenoxyacetate (2,4,5-T), an herbicide and chlorinated aromatic compound. Although some progress has been made in understanding 2,4,5-T degradation by AC1100 by molecular analysis, little is known about the biochemistry involved. Enzymatic activity converting 2,4,5-T to 2,4,5-trichlorophenol in the presence of NADH and O(inf2) was detected in cell extracts of AC1100. Phenyl agarose chromatography of the ammonium sulfate-fractionated cell extracts yielded no active single fractions, but the mixing of two fractions, named component A and component B, resulted in the recovery of enzyme activity. Component B was further purified to homogeneity by hydroxyapatite and DEAE chromatographies. Component B had a native molecular weight of 140,000, and it was composed of two 49-kDa (alpha)-subunits and two 24-kDa (beta)-subunits. Component B was red, and its spectrum in the visible region had maxima at 430 and 560 nm (shoulder), whereas upon reduction it had maxima at 420 (shoulder) and 530 nm. Each mole of (alpha)(beta) heterodimer contained 2.9 mol of iron and 2.1 mol of labile sulfide. These properties suggest strong similarities between component B and the terminal oxygenase components of the aromatic ring-hydroxylating dioxygenases. Component A was highly purified but not to homogeneity. The reconstituted 2,4,5-T oxygenase, consisting of components A and B, converted 2,4,5-T quantitatively into 2,4,5-trichlorophenol and glyoxylate with the coconsumption of NADH and O(inf2).     

甘肃莲花山及其邻近地区大型真菌调查初报

本文报道了甘肃莲花山及其邻近地区的大型真茵１１７种,根据Ａｉｎｓｗｏｒｔｈ,Ｇ．Ｃ．系统（１９７３）,隶属于６０屑,２６科,２亚门,其中食用菌７０种,药用菌３７种,毒蘑菇２３种,菌根菌２４种,甘肃分布新纪录５０种。     

Purification and characterization of a tetrachloro-p-hydroquinone reductive dehalogenase from a Flavobacterium sp.

Tetrachloro-p-hydroquinone (TeCH) is the first intermediate in pentachlorophenol (PCP) degradation by Flavobacterium sp. strain ATCC 39723. We previously purified a PCP hydroxylase that oxidized PCP to TeCH. Subsequently, we identified the reductive dehalogenation of TeCH to 2,3,6-trichloro-p-hydroquinone and then to 2,6-dichloro-p-hydroquinone in a cell extract with the reduced form of glutathione as the reducing agent under anaerobic conditions. Here we report the purification of a TeCH reductive dehalogenase that reductively dehalogenated TeCH to trichlorohydroquinone and then to dichlorohydroquinone. The enzyme was purified by protamine sulfate treatment, ammonium sulfate fractionation, and phenyl-agarose, anion-exchange, and gel filtration column chromatographies. As determined by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses, the protein has a molecular weight of about 30,000; nondenaturing polyacrylamide gel electrophoresis analysis suggests that the native enzyme exists as a dimer. The enzyme used glutathione but not NADPH, NADH, dithiothreitol, or ascorbic acid as the reducing agent. The optimal pH was close to neutral.     

Regulation of synthesis and activity of NAD(+)-dependent 15-hydroxy-prostaglandin dehydrogenase (15-PGDH) by dexamethasone and phorbol ester in human erythroleukemia (HEL) cells

Dexamethasone stimulated 15-PGDH activity in HEL cells in a time and concentration dependent manner. Increase in 15-PGDH activity by dexamethasone was found to be accompanied by an increase in enzyme synthesis as revealed by Western blot and [35S]methionine labeling studies. In addition to dexamethasone, other anti-inflammatory steroids also increased 15-PGDH activity in the order of their glucocorticoid activity. Among sex steroids only progesterone increased significantly 15-PGDH activity. 12-0-Tetradecanoylphorbol-13-acetate (TPA) also induced the synthesis of 15-PGDH but inhibited the enzyme activity. It appears that TPA caused a time dependent inactivation of 15-PGDH by a protein kinase C mediated mechanism.     

Biodegradation of triiodophenol by cell-free extracts of a pentachlorophenol-degrading Flavobacterium sp

Pentachlorophenol (PCP) degrading Flavobacterium sp. ATCC 39723 was found to degrade other polyhalogenated phenolic compounds, including triiodophenol, tribromophenol, and trichlorophenol. Each compound was able to induce the degradation of the other compounds. A PCP Flavobacterium sp. mutant, F-2, was unable to degrade any of the halogenated compounds. The results suggest that all of the polyhalogenated phenols were degraded by the same enzyme system. This observation led us to exploit the sensitive leuco crystal violet assay, which measures the iodide released from triiodophenol. Cell free extracts from PCP-induced cells were able to release iodide from triiodophenol. The reaction required NADPH and oxygen.     

杭州双睾虫生活史各阶段体壁的透射电镜观察

作者应用透射电镜技术观察了杭州双睾虫(Diplorchis hangzhouensis)生活史各阶段(自由生活的钩毛蚴、蝌蚪鳃上的后期幼虫、沼蛙膀胱内的成虫)体壁的超微结构,讨论了幼虫发育至成虫过程中这些结构演变的意义,并与其它单殖吸虫的体壁进行了比较。     

赤眼蜂的产卵行为

赤眼蜂能在多种宿主卵中产卵寄生,并且还具有在人工物体上产卵的习性。Holloway早在1912年即观察到秋葵植物汁滴干燥结膜后,广赤眼蜂(Trichogramma evanescen)企图在其表面产卵。Salt(1935)观察到赤眼蜂有企图将产卵器插入玻璃珠,水银滴,植物种子等表面的行为。Hagen等(1965)试制人工蜡卵饲喂草蛉幼虫时,偶然发现赤眼蜂可以产卵在这种蜡卵内。近年来,Rajendram和Hagen(1974)曾用上述蜡卵引诱赤眼蜂产卵于盐水或氨基酸溶液内。总之,赤眼蜂可在不少的非天然的宿主或物体上产卵,并且所报道产过卵的物体都是球形的。Salt(1937)认为决定赤眼蜂产卵的因素是化学感觉,夜蛾中促使赤眼蜂产卵的物质是22—25碳的直链烷烃成分。但影响产卵行为的各种因素尚缺乏详细的分析。     

Tumour‐associated macrophages as a novel target of VEGI‐251 in cancer therapy

Tumour‐associated macrophages (TAMs), which possess M2‐like characters and are derived from immature monocytes in the circulatory system, represent a predominant population of inflammatory cells in solid tumours. TAM infiltration in tumour microenvironment can be used as an important prognostic marker in many cancer types and is a potential target for cancer prevention or treatment. VEGI‐251 not only is involved in the inhibition of tumour angiogenesis, but also participates in the regulation of host immunity. This work aimed to investigate the involvement of VEGI‐251 in the regulation of specific antitumour immunity. We found that recombinant human VEGI‐251(rhVEGI‐251) efficiently mediated the elimination of TAMs in tumour tissue in mice, and induced apoptosis of purified TAMs in vitro. During this process, caspase‐8 and caspase‐3 were activated, leading to PARP cleavage and apoptosis. Most importantly, we further elucidated the mechanism underlying VEGI‐251‐triggered TAM apoptosis, which suggests that ASK1, an intermediate component of the VEGI‐251, activates the JNK pathway via TRAF2 in a potentially DR3‐dependent manner in the process of TAM apoptosis. Collectively, our findings provide new insights into the basic mechanisms underlying the actions of VEGI‐251 that might lead to future development of antitumour therapeutic strategies using VEGI‐251 to target TAMs.     

不同肝血流阻断方式肝切除手术对原发性肝癌合并肝硬化患者肝功能及肠黏膜屏障的影响

目的:探讨肝切除手术运用Pringle法阻断、半肝血流阻断(HVC)后对原发性肝癌合并肝硬化患者肝功能及肠黏膜屏障的影响。方法:选取2016年4月～2019年9月期间我院收治的原发性肝癌合并肝硬化患者93例,根据随机数字表法将患者分为A组(n=46,Pringle法阻断)和B组(n=47,HVC),比较两组患者围术期指标、肝功能指标[谷丙转氨酶(ALT)、谷草转氨酶(AST)以及总胆红素(TBIL)]、肠黏膜屏障指标[D-乳酸,内毒素]及并发症发生情况。结果:两组阻断时间、术中失血量、手术时间比较无差异(P0.05);B组住院时间短于A组(P0.05)。两组术前、术后3 d、术后7 d ALT、AST、TBIL呈升高后降低趋势,且B组低于A组(P0.05)。两组患者术后并发症发生率比较无差异(P0.05)。两组术前、术后3 d、术后7dD-乳酸、内毒素呈升高后降低趋势,且B组低于A组(P0.05)。结论:与Pringle法阻断相比,原发性肝癌合并肝硬化患者在肝切除手术中运用HVC,可有效缩短住院时间,减轻肝功能及肠黏膜屏障损害,且不增加并发症发生率,临床应用价值较高。