Toxic effects of Fusarium mycotoxin butenolide on rat myocardium and primary culture of cardiac myocytes.

Mycotoxin toxicosis has been implicated in the etiopathogenesis of Keshan disease (KD), an endemic cardiomyopathy prevailing in some regions of China. Butenolide (4-acetamido-4-hydroxy-2-butenoic acid gamma-lactone, CAS No. 16275-44-8), a mycotoxin produced by several Fusarium species such as Fusarium tricinctum and Fusarium graminearum, is frequently detected from the cereals in the endemic areas of KD. The present study is undertaken to investigate whether this mycotoxin can induce myocardial damage. Exposure of primary culture of cardiac myocytes to butenolide resulted in significant cytotoxicity, manifested by changes in cell morphology and decreases in cell viability. Consistent with the in vitro findings, distinct myocardial toxicity in vivo was observed after administration of rats by gavage with butenolide (10 and 20 mg/kg/day) for 2 months, and the myocardial injuries were characterized by focal necrosis of myocardium and fragmentation of myofiber. Butenolide also induced significant oxidative damage to the myocardium in vitro evidenced by a concentration-dependent lipid peroxidation in the myocardial homogenates, whereas antioxidants superoxide dismutase (SOD), N-acetylcysteine (NAC) and glutathione (GSH) provided significant protections against this oxidative effect. Taken together, these results clearly reveal that butenolide possesses the potential to induce myocardial toxicity. The present findings may reinforce the hypothesis that toxicosis by mycotoxins is one of the etiological factors for KD.     

苦参液雾化吸入辅助治疗小儿哮喘

目的:观察苦参液雾化吸入辅助治疗小儿哮喘的临床疗效.方法:本组88例,随机分成中西药治疗组及西药对照组,每组44例,对照组常规用抗生素、激素及口服美喘清;治疗组常规用抗生素、激素,并用苦参液雾化吸入.结果:治疗组44例,哮喘症状及体征消失时间均短于对照组(p＞0.05).结论:苦参液雾化可作为小儿哮喘的重要辅助治疗方法之一.     

Expression, purification and molecular modeling of recombinant fibrinogenase [IV], a metalloproteinase from Deinakistrodon acutus venom.

A novel metalloproteinase, recombinant fibrinogenase IV (rFIV(a)), was expressed and purified from Deinakistrodon acutus venom. It was a single chain protein with an apparent molecular weight 27 kDa and an isoeletric point of pH 7.1. RFIV(a) cleaved preferentially the Aalpha-chain and also cleaved Bbeta, gamma-chains of fibrinogen when the incubation time was prolonged. The proteolytic activity was inhibited by EDTA, l-cysteine, and DTT, indicating rFIV(a) was a metalloproteinase requiring disulfide bonds for its activity. It kept above 85% of the initial activity from pH 4.5-11, showed an equal maximum activity at the temperature range from 30 to 50 degrees C, and was inactivated by Zn2+, Cu2+ and Cd2+. Homology modeling of rFIV(a) showed that two highly conserved disulfide bonds (Cys159-Cys164 and Cys117-Cys197) was maintained from its structure, and it exhibited the characteristic conserved motif H142E143XXH146XXGXXH152, whose three histidine residues were involved in binding of the catalytically essential zinc ion. This work demonstrates the expression, purification and characterization of recombinant fibrinogenase IV, which belongs to class P-I metalloproteinase from D. acutus venom.     

复方葡萄籽提取物对乙醇及四氯化碳肝损伤的影响

目的：探讨复方葡萄籽提取物（GSE）对化学性肝损伤的预防和治疗作用。方法：用小鼠建造四氯化碳肝损伤和乙醇肝损伤模型，观察乙醇灌胃前预先给予复方GSE及乙醇或四氯化碳灌胃后再给予复方GSE后小鼠血清谷丙转氨酶（ALT），肝细胞超氧化物歧化酶（SOD）、谷光甘肽（GSH）、丙二醛（MDA）。结果：预先给予GSE可增强SOD活性，防止乙醇导致的肝GSH减少，有效降低小鼠血清ALT和肝细胞MDA上升程度，在乙醇或四氯化碳灌胃后再给予复方GSE，也可降低小鼠血清中ALT和肝细胞脂质过氧化产物。结论：复方GSE对乙醇肝损伤有预防和治疗作用。对四氯化碳肝损伤也有一定治疗作用。     

实验性隐睾术后大鼠附睾的反应性变化

本研究描述了实验性隐睾术后大鼠附睾的形态学变化。结果表明,隐睾术后由于腹温作用和附皋管管腔内容物的改变,附睾管上皮细胞的形态和功能明显改变。术后早期上皮细胞出现无丝分裂相,上皮细胞的分泌和吞噬功能加强,胞质内含PAS阳性颗粒的细胞增多。随术后时间的延长,附睾上皮细胞变化逐渐趋于稳定。[     

Longitudinal Analysis of Severe Acute Respiratory Syndrome (SARS) Coronavirus-Specific Antibody in SARS Patients

The serum antibodies to severe acute respiratory syndrome (SARS) coronavirus of 18 SARS patients were checked at 1 month and every 3 months after disease onset. All of them except one, who missed blood sampling at 1 month, tested positive for the immunoglobulin G (IgG) antibody at 1 month. Fifteen out of 17 tested positive for the IgM antibody at 1 month. The serum IgM antibody of most patients became undetectable within 6 months after the onset of SARS. The IgG antibody of all 17 patients, whose serum was checked 1 year after disease onset, remained positive.     

Flavonoids modulate monocarboxylate transporter-1-mediated transport of gamma-hydroxybutyrate in vitro and in vivo.

The objective of this study was to determine the effects of flavonoids on the in vitro monocarboxylate transporter 1 (MCT1)-mediated transport and in vivo disposition of the drug of abuse, gamma-hydroxybutyrate (GHB). The uptake of GHB in rat MCT1 gene-transfected MDA-MB231 cells was significantly decreased in the presence of the flavonoids apigenin, biochanin A, chrysin, diosemin, fisetin, genistein, hesperitin, kaempferol, luteolin, morin, narigenin, phloretin, and quercetin, but was not affected by the flavonoid glycosides phloridzin and rutin. The IC(50) values for luteolin, morin, and phloretin were 0.41 +/- 0.14, 6.41 +/- 2.01, and 2.57 +/- 0.48 microM, with the inhibition mechanism for luteolin being competitive. [(3)H]Kaempferol and [(3)H]biochanin A did not exhibit MCT1-mediated uptake, suggesting that these flavonoids are not substrates for MCT1. The combination of luteolin and phloretin inhibited the uptake of GHB in a synergistic manner; however, the combination of luteolin and morin was antagonistic. GHB 1000 mg/kg was administered to rats by i.v. bolus, with or without the concomitant administration of luteolin 10 mg/kg i.v. After luteolin treatment, the renal and total clearances of GHB were significantly increased, probably because of inhibition of the MCT1-mediated renal reabsorption of GHB, and the sleep time significantly decreased (121 +/- 5 min versus 165 +/- 10 min) compared with control rats. Overall, the results of this study indicate that flavonoids from food or herbal products may significantly alter the pharmacokinetics and pharmacodynamics of MCT substrates.     

慢性肾功能不全患者血管活性物质测定在血液净化中的临床研究

目的 评价血管紧张素Ⅱ、氨甲酰血红蛋白、醛固酮及甲状旁腺激素等在血液净化充分性中的意义。方法 120例慢性肾功能衰竭患者，按采用不同的透析膜分为3组：即聚砜膜、醋酸膜、铜仿膜。用放射免疫法测定其血管紧张素Ⅱ（AngⅡ）及醛固酮（ALD）浓度，化学发光法测甲状旁腺激素（PTH）。采用不同的血流量200ml／min及250ml／min比较。结果 对体内中、小分子物质清除率，聚砜膜〉醋酸膜〉铜仿膜。结论 高血流量、高通透性人工合成聚砜膜，对治疗慢性肾功能衰竭，提高肾功能不全患者的身心健康及生活质量、延长患者生命有很重要的临床意义。     

Comparative expression profiling in primary and immortalized endothelial cells: changes in gene expression in response to hydroxy methylglutaryl-coenzyme A reductase inhibition.

Immortalized cell lines offer significant logistical advantages over primary cells when used for in-vitro studies. Immortalized cells may, however, exhibit important differences relative to their primary cell counterparts. In this study, microarrays were used to make a genome-wide comparison between primary human umbilical vein endothelial cells (HUVECs) and EA.hy926, an immortalized HUVEC cell line, in their baseline properties and in their response to inhibition of the mevalonate pathway with an inhibitor of hydroxy methylglutaryl-coenzyme A reductase (statin). HUVECs and EA.hy926 were incubated with control medium, atorvastatin, mevalonate, or a combination of atorvastatin and mevalonate for 24 h. Gene expression profiles were obtained in duplicates using Affymetrix Human Genome U133A 2.0 arrays (Santa Clara, California, USA). Probe-sets were selected according to the following criteria: a twofold or greater increase/decrease in atorvastatin-treated cells compared with untreated cells; a twofold or greater reversal of the effect of atorvastatin by combined treatment with atorvastatin and mevalonate; no significant change in gene expression in cells treated with mevalonate alone compared with untreated cells. Most genes that were expressed by untreated HUVECs, were also expressed by untreated EA.hy926 cells. EA.hy926 cells, however, constitutively expressed a large number of additional genes, many of which were related to cell cycle control and apoptosis. Atorvastatin induced differential expression (> or = twofold) of 103 genes in HUVECs (10 up, 93 down) and 466 genes in EA.hy926 cells (198 up, 268 down). Applying the above selection criteria, thrombomodulin and tissue plasminogen activator were up-regulated in both cell types, whereas, connective tissue growth factor, thrombospondin-1, and cysteine-rich angiogenic inducer 61 were down-regulated. In conclusion, EA.hy926 cells retain most of the characteristics of endothelial cells under baseline conditions as well as after treatment with atorvastatin. It is necessary, however, to carefully select and validate changes in genes that are the focus of studies when using EA.hy926 cells. While this cell line is highly useful in studies on some genes, including genes encoding molecules involved in regulating thrombohemorrhagic homeostasis, they appear to be less suited for studies focused on other genes, particularly those involved in the regulation of cell proliferation and apoptosis.     

丝裂霉素C与C2-神经酰胺联合应用治疗膀胱癌

目的 观察丝裂霉素C（MMC）与C2-神经酰胺（C2-cer）联合应用对人膀胱癌细胞的作用效果，并探讨其机制。方法 不同浓度MMC与C2-cer单独及联合作用于人膀胱癌BIU-87细胞后，应用噻唑蓝（MTT）比色法检测细胞生长抑制率，计算合用指数（CI），流式细胞仪（FCM）检测BIU-87细胞凋亡率，吖啶橙（AO）荧光染色观察凋亡形态学变化，Western blot检测细胞色素C在细胞内分布变化，并检测Caspase-3活性改变。结果 单独应用时MMC与C2-cer的中效浓度分别是159和28μmol／L，联合用药时下降为55和11μmol／L，CI=0．74。MMC与C2-cer单独及联合应用均可导致BIU-87细胞出现凋亡的形态学变化。两种药物联合应用时的凋亡率高于各自单用（P＜0．05）。线粒体细胞色素C含量在MMC与C2-cer单独及联合应用时均较对照组减少，联合用药时减少最为明显，细胞质内细胞色素C含量在联合用药时增加也最为明显（P＜0．05）。Caspase-3活性在MMC与C2-cer单独及联合应用时均较对照组升高，联合用药时升高最为明显（P＜0．05）。结论 MMC与C2-cer联合应用可以通过共同诱导细胞凋亡，协同抑制膀胱癌细胞生长。线粒体细胞色素C释放和Caspase-3活性变化可能发挥重要作用。