Bis‐2(5H)‐furanone derivatives as new anticancer agents: Design,synthesis, biological evaluation,and mechanism studies

New bis‐2(5H)‐furanone derivatives containing a benzidine core were synthesized via a one‐step transition‐metal‐free reaction of benzidine with 5‐substituted 3,4‐dihalo‐2(5H)‐furanones. Their antitumor activities against various tumor cells have been evaluated by MTT assay. Among them, compound 4e exhibits significant inhibitory activity against C6 glioma cells with an IC₅₀ value of 12.1 μm and low toxicity toward HaCaT human normal cells. Studies on the antitumor mechanism reveal that cell cycle arrest at S‐phase in C6 cells is induced by compound 4e . Furthermore, investigations with electronic, fluorescence emission and circular dichroism spectra show that compound 4e can significantly interact with C6‐DNA. These data indicate that DNA may be one of the potential targets for bis‐2(5H)‐furanone derivatives as anticancer drugs.     

两种大便隐血试验测定方法的比较

粪便隐血试验是临床诊断消化道出血性疾病、筛查消化道肿瘤的一项重要常规检查项目之一.化学法由于费用低廉、反应快速、结果容易判断等优点在很多医院长期使用,但容易受饮食,一些化学药品、铁剂影响出现假阳性或假阴性结果.用免疫胶体金法检测粪便隐血,有较高的敏感性和特异性.本文对联苯胺法和免疫胶体金法两种方法测定大便隐血试验进行了比较,二者差异有统计学意义.     

Antioxidant activities of curcumin and combinations of this curcuminoid with other phytochemicals

Biomedical investigations of curcumin (and curcuminoids) have provided evidence of a wide range of molecular and cellular activities, most related to redox reactions and signal transduction. The main goal of the present study was to compare antioxidant activities of curcumin with those of resveratrol, a polyphenol present in some dietary plants such as Vitis vinifera (L.) and Arachis hypogaea (L.) and many other, non‐dietary plants. Combinations of the two were also examined for potential synergism in a heme‐enhanced oxidation reaction. Curcumin exhibited antioxidant effects at all time points (1–5 min; 10 μM), e.g., 30.5 ± 11.9% (SEM) oxidation relative to controls without phytochemicals (p < 0.01) at 3 min, a time chosen for comparisons. The same concentration of resveratrol exhibited about half of curcumin's activity. Curcumin and resveratrol together (5 μM each) resulted in a synergistic antioxidant effect: 15.5 ± 1.7% greater than an average of individual activities. This synergy was significantly greater (p < 0.05; about 4‐fold) than that of curcumin together with the flavonol quercetin. In conclusion, curcumin is a potent antioxidant in a reaction that may be relevant to in vivo toxicity. In relation to two other well‐known antioxidants, curcumin shows significantly greater synergism with resveratrol than with quercetin. Copyright © 2009 John Wiley & Sons, Ltd.     

Hypochlorous acid-mediated activation of N-acetylbenzidine to form N'-(3'-monophospho-deoxyguanosin-8-yl)-N-acetylbenzidine.

Hypochlorous acid (HOCl), a chemically reactive oxidant, is an important component of the inflammatory response and may contribute to carcinogenesis. This study assessed the possible activation of N-acetylbenzidine (ABZ) by HOCI to form a specific DNA adduct, N'-(3'-monophospho-deoxyguanosin-8-yl)-N-acetylbenzidine. HOCl was incubated with 0.06 mM 3H-ABZ, and transformation assessed by HPLC. Similar results were observed at pH 5.5 or 7.4. A linear increase in transformation was observed from 0.025 to 0.1 mM HOCl with up to 80% of ABZ changed. Approximately, 2 nmoles of HOCI oxidized 1 nmole of ABZ. N-oxidation products of ABZ metabolism, such as N'-hydroxy-N-acetylbenzidine, were not detected. Oxidation of ABZ was prevented by taurine, DMPO, glutathione, and ascorbic acid, whereas mannitol was without effect. Results are consistent with a radical mechanism. In the presence of 2'-deoxyguanosine 3'-monophosphate (dGp), a new product (dGp-ABZ) was observed. The same adduct was observed with DNA. dGp-ABZ was found to be quite stable (>80% remaining) at 70 degrees C in pH 5.5 (60 min) and 7.4 (240 min). Electrospray mass spectrometry indicated that dGp-ABZ was N'-(3'-monophospho-deoxyguanosin-8-yl)-N-acetylbenzidine, and this was confirmed by NMR. 32P-postlabeling in combination with TLC and HPLC determined that the adduct made by either HOCl or prostaglandin H synthase oxidation of ABZ in the presence of dGp or DNA was dGp-ABZ. Thus, HOCI activates ABZ to form dGp-ABZ and may be responsible for the presence of this adduct in peripheral white blood cells from workers exposed to benzidine. Reaction of ABZ with HOCl provides an easy, convenient method for preparing dGp-ABZ.     

葛根素、川芎嗪、丹参注射液对大脑中动脉阻断大鼠脑微循环血流量的影响

目的：探讨葛根素、川芎嗪、丹参注射治疗脑血管疾病的机理。方法：采用穿线法阻断大脑中动脉（ＭＣＡＯ）复制大鼠脑缺血模型 循环动态分析仪,观察上述药物对脑微循环灌流量的下降至结扎前的１７％左右,各用药组用后微循环血流量均较ＭＣＡＯ组是升高,其中丹参了为显著。结论：葛根素、川芎嗪１丹参可明显改善ＭＣＡＯ后脑微循环血流量,是其治疗脑血管疾病的重要机理之一。     

A new brain cell surface glycoprotein identified by monoclonal antibody

Of 207 monoclonal antibodies produced against cultured mouse cerebellar cells, 16 reacted with cerebellar cell surfaces and 4 reacted with glycoproteins. One of them, called anti-BSP-3 (Brain cellSurfaceProtein-3) defines a 48,000 molecular weight protein which can be iodinated at the surface of cultured cerebellar cells. Lectin-binding and sugar incorporation studies established the glycoprotein nature of the antigen. Astroglia (glial fibrillary acidic protein-positive cells) in primary cerebellar cultures were labelled intensely for this antigen by the indirect immunofluorescence method while neuronal cells and their processes were more weakly labelled. Fibronectin-positive cells were negative for BSP-3. In cerebellar sections using the immunoperoxidase method at both the optical and electron microscope levels, the difference in staining intensity between astrocytes and neuronal cells was not significant: in Purkinje cells and in the large neurones present in the deep cerebellar nuclei the immunoperoxidase percipitate was confined to the plasma membrane while in both astrocytes and granule cells cytoplasmic labelling was also observed. Oligodendrocytes do not appear to react with the anti-BSP-3 monoclonal antibody; neither do endothelial or leptomeningeal cells.The availability of a monoclonal antibody produced by a stable hybridoma line will be a powerful tool in attempts to purify the BSP-3 antigen and to elucidate its function.     

联苯胺亲和层析纯化尿激酶

利用甲基丙烯酸缩水甘油酯与乙二醇甲基丙烯酸酯经悬浮聚合反应合成亲水性亲和载体骨架，骨架上的不环氧基与氨水反应，继之与丁二酸酐反应，最后与配基联苯胺偶联制备亲和载体，配基密度为５４μｍｏｌ／ｇ湿胶。亲和载体吸附激酶的最佳条件为ＰＨ４．６和２．５ｍｏｌ／ＬＮａＣｌ；测定了吸附等温线，算得离常数Ｋｄ为３１６ＩＵ／ｍｌ，了大吸附量ｑｍ为２．３８３×１０＾４Ｕ／ｍＬ湿胶，洗脱条件为含０．５ｍｏｌ／ＬＮａＣｌ     

Expression of visfatin in alloxan-induced diabetic rat testis

Diabetes mellitus is a potential epidemic all over the world and causes dysfunction of reproductive activity. Visfatin, one of the adipokines, is present in various tissues including the testis. Our hypothesis was the level of testicular visfatin is affected in diabetic condition. The aim of the present study was to investigate the expression and localization of visfatin in the diabetic rat testis. No similar studies have been performed in diabetic rat testis with reference to visfatin. Overnight fasted adult male Wistar rats were made diabetic by the administration of alloxan (150 mg/kg i.p., in 0.9% saline). Blood glucose levels were tested on five days after alloxan treatment, rats with high blood glucose levels (>250 mg/dL) were considered as diabetic. Immunolocalization and Western blotting analysis of visfatin were performed. Correlation of visfatin expression was made in relation to body weight, testis weight, glucose concentration and serum testosterone level. Expression of visfatin was observed in Leydig cells, spermatocytes and sperm in control as well as in the diabetic group. Mild immunostaining of visfatin was observed in affected seminiferous tubules of alloxan-induced diabetic rat testis. Western blot analysis showed decreased expression of testicular visfatin in diabetic rats. The expression of visfatin showed a positive correlation with serum testosterone levels, body and testis weight, while a negative correlation was observed with blood glucose levels. This study showed involvement of visfatin in diabetic associated impairment of testicular activity.     

Re-evaluation of the latent period of bladder cancer in dyestuff-plant workers in Japan

BACKGROUND: The carcinogenesis of benzidine (BZ) and beta-naphthylamine (BNA) for bladder is well known. Although it was thought to be rare to develop occupational bladder cancer more than 20 years after the exposure to these chemicals, there are still new clinical cases even 30 years after exposure. The purpose of this study was to re-evaluate the latent carcinogenic period of BZ and BNA, in order to set the safety period after exposure for the health surveillance system. METHODS: The subjects were 236 dyestuff-plant workers in Tokyo, who had been exposed to these dyestuffs. The incidence of bladder cancer and its histopathology in this group was surveyed in the period from 1962 to 1996. RESULTS: Nineteen workers (8.1%) were found to have bladder cancers. The exposure period for these 19 patients was 82.0 +/- 50.2 months. The mean +/- SD latent period from the subjects' initial and final exposure until tumor development was 29.5 +/- 8.2 years and 20.1 +/- 10.6 years, respectively. Significantly, a negative correlation (Pearson) was observed between the exposure period and the latent period from the end of exposure to cancer onset (R = -0.544, P < 0.05). All tumors except one were transitional cell carcinoma. Flow cytometric analysis was performed in 11 patients and all of these patients had DNA aneuploidy. CONCLUSIONS: The latent periods of bladder cancer caused by BZ and BNA were longer than previously expected. It is necessary to survey the onset of bladder cancer in exposed workers more than 30 years after the initial exposure.     

Cytoarchitectonic organization of laryngeal motoneurons within the nucleus ambiguus of the cat

The central distribution of laryngeal motoneurons was studied in the cat by retrograde axonal transport of horseradish peroxidase. The enzyme was injected selectively into the cricothyroid (CT), lateral cricoarytenoid and thyroarytenoid (LCA-TA), and posterior cricoarytenoid (PCA) muscles of the larynx with or without the previous sectioning of the left laryngeal recurrent nerve (LR) or the left superior laryngeal nerve (SL). The CT motoneurons appeared as a compact group of medium-size cells located in the rostral one-third of the nucleus ambiguus (nA). The LCA-TA motoneurons were found in the caudal two-thirds of the nA, constituting a loose group of large motoneurons. The PCA motoneurons were located throughout the whole extend of the nA, the cells being large in the caudal pole and smaller in the rostral one-third of nA. Laryngeal muscle innervation was exclusively of ipsilateral origin. Axonal projections in the brain stem were different depending on the nerve (LR or SL) by which the efferent fibers were sent.