芍药甙诱导人肝癌细胞系HepG2凋亡的研究

目的：观察芍药甙诱导人肝癌细胞系HepG2凋亡及凋亡相关基因的变化。方法：用0．5～0．2g／L芍药甙处理人肝癌HepG2细胞，光镜下观察细胞形态；MTT法检测细胞增殖率；流式细胞仪检测凋亡峰；免疫组化法检测P53，bcl-2，bax等凋亡相关基因的表达。结果：0．5～2．0g／L浓度的芍药甙对HepG2有显著抑制作用；可诱导HepG2凋亡，并能上调凋亡相关基因P53和bax表达、下调抗凋亡基因bcl-2表达。结论：0．5～2．0g／L芍药甙可诱导人肝癌细胞HepG2凋亡，其机制可能与凋亡相关基因表达增强和抗凋亡相关基因表达减弱有关。     

乙醇对原代培养大鼠肝细胞损伤的体外研究

随着乙醇消耗的增多,乙醇对肝脏的损害日渐突出,但乙醇性肝损伤的机制仍不十分清楚,为此,建立乙醇性肝损伤细胞培养模型,进一步从细胞水平研究乙醇性肝损伤的发病机制。     

芍药甙对急性坏死性胰腺炎大鼠NF-κBp65及细胞因子表达的影响

目的观察芍药甙对ANP大鼠的治疗效果及对NF-κB、TNF-α、IL-6表达的影响。方法40只大鼠随机分为正常组、ANP 24h、72h组和芍药甙24h、72h治疗组(芍药组),每组8只。采用L -精氨酸腹腔内注射法诱发ANP模型,芍药组于造模后即刻用2ml芍药甙药液灌胃,11次/2h。各组采血检测血清淀粉酶,取胰腺组织行病理检查,Western blot法检测NF-κB p65蛋白表达,实时荧光定量PCR法检测TNF-αmRNA、IL-6 mRNA的表达。结果造模后24h ANP组和芍药组血清淀粉酶分别为(3569±516)U/L和(2994±370)U/L,明显高于正常组的(1136±107)U/L(P<0.05);病理学分值分别为4.25±0.63和3.06±0.49,也明显高于正常组的0分(P<0.05);芍药组24h的NF-κB p65蛋白及TNF-αmRNA、IL-6 mRNA表达相对值分别为0.230±0.044,0.141±0.018和0.017±0.029,72h分别为0.040±0.006、0.078±0.017和0.008±0.001,均显著低于同时间点的ANP组(P<0.05)。结论芍药甙能减轻ANP大鼠胰腺的病理损害程度,其机制可能与抑制NF-κB活化、抑制TNF-αmRNA、IL-6 mRNA表达有关。     

芍药甙对急性坏死性胰腺炎大鼠NF-kB p65及细胞因子表达的影响

目的 观察芍药甙对ANP大鼠的治疗效果及对NF-kB、TNF-α、IL-6表达的影响.方法 40只大鼠随机分为正常组、ANP24h、72h组和芍药甙24h、72h治疗组(芍药组),每组8只.采用L-精氨酸腹腔内注射法诱发ANP模型,芍药组于造模后即刻用2ml芍药液灌胃,11次/2h.各组采血检测血清淀粉酶,取胰腺组织行病理检查,Western blot 法检测 NF-kB p65蛋白表达,实时荧光定量PCR法检测TNF-α mRNA\IL-6 mRNA的表达.结果 造模后24h ANP组和芍药血清淀粉酶分别为(3569±516)U/L和(2994±370)U/L,明显高于正常组的(1136±107)U/L(P＜0.05);病理学分值分别为4.25±0.63和3.06±0.46,也明显高于正常组的0分(P＜0.05);芍药组24h的NF-kB p65蛋白及TNF-αmRNA、IL-6 mRNA表达相对值分别为0.230±0.0440.141±0.018和0.017±0.029,72h分别为0.040±0.006、0.078±0.017和0.008±0.001,均显著低于同时间点的ANP组(P＜0.05).结论芍药甙能减轻ANP大鼠胰的病理损害程度,其机制可能与抑制NF-kB活化、抑制TNF-α mRNA、IL-6 mRNA表达有关.     

芍药甙对急性坏死性胰腺炎大鼠NF-kB p65及细胞因子表达的影响

目的 观察芍药甙对ANP大鼠的治疗效果及对NF-kB、TNF-α、IL-6表达的影响.方法 40只大鼠随机分为正常组、ANP24h、72h组和芍药甙24h、72h治疗组(芍药组),每组8只.采用L-精氨酸腹腔内注射法诱发ANP模型,芍药组于造模后即刻用2ml芍药液灌胃,11次/2h.各组采血检测血清淀粉酶,取胰腺组织行病理检查,Western blot 法检测 NF-kB p65蛋白表达,实时荧光定量PCR法检测TNF-α mRNA\IL-6 mRNA的表达.结果 造模后24h ANP组和芍药血清淀粉酶分别为(3569±516)U/L和(2994±370)U/L,明显高于正常组的(1136±107)U/L(P＜0.05);病理学分值分别为4.25±0.63和3.06±0.46,也明显高于正常组的0分(P＜0.05);芍药组24h的NF-kB p65蛋白及TNF-αmRNA、IL-6 mRNA表达相对值分别为0.230±0.0440.141±0.018和0.017±0.029,72h分别为0.040±0.006、0.078±0.017和0.008±0.001,均显著低于同时间点的ANP组(P＜0.05).结论芍药甙能减轻ANP大鼠胰的病理损害程度,其机制可能与抑制NF-kB活化、抑制TNF-α mRNA、IL-6 mRNA表达有关.     

雷公藤多甙防治大鼠血管球囊损伤后平滑肌细胞增殖的实验研究

目的观察不同剂量雷公藤多甙对颈动脉球囊损伤后大鼠血管平滑肌细胞增殖的影响,探讨其在再狭窄防治方面的作用。方法SD雄性大鼠32只,随机分为4组。取8只为正常组,其余24只以内皮剥脱法造模,随机分为模型组、雷公藤多甙小剂量组及大剂量组各8只。模型组术后予蒸馏水3mL/(kg·d)灌胃,小剂量组予雷公藤多甙30mg/(kg·d)灌胃,大剂量组予雷公藤多甙60mg/(kg·d)灌胃。于术前处死8只正常组大鼠,于术后2周处死各实验组动物。取材检测指标:HE染色观察管壁增生情况,计算机图像分析内、中膜厚度及其比值,免疫组化法检测增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)表达水平。结果雷公藤小剂量组内膜、中膜厚度与内膜/中膜厚度值分别为(54.19±11.59)μm、(102.66±15.58)μm与0.55±0.18;雷公藤大剂量组各指标分别为(37.58±8.51)μm、(115.09±18.98)μm与0.33±0.08。抑制PCNA表达,升高α-SMA水平,并呈剂量正相关。结论雷公藤多甙能有效抑制血管损伤后内膜增生,对大鼠血管球囊损伤后的狭窄有一定的防治作用。     

脾气虚大鼠肝细胞超微结构动态变化的实验研究

用过劳及饮食失节法塑造大鼠气虚模型，结果表明，随着造模时间的延长，脾气虚大鼠肝细胞线粒体的肿胀、基质变浅、空泡变淡、嵴断裂及排列紊乱等逐中重，同时空肠细胞线粒体在似变化，提示脾气虚大鼠肝细胞线粒体的变化是一个由轻到重的渐进过程，其病变程度与空脾性细胞线粒体的病变程度密切相关；随着营养物质吸收障碍的逐渐加重，肝细胞线粒体的病变程度也渐趋严重，肝脏合成血浆蛋白的能力逐渐下降，是脾气虚证低蛋白血症逐渐加     

氟对大鼠肝细胞影响及SOD阻断因子的透射电镜研究

本实验分正常组、投氟组、投氟加超氧化物歧化酶(SOD)组喂养大鼠六个月后处死,制作肝组织的透射电子显微镜标本,进行超微结构观察。结果发现,投氟组肝细胞产生明显的毒性作用,表现为肝细胞肿胀,细胞膜破损,线粒体变性,核形不整、固缩,一些肝细胞崩解。在投氟加 SOD 的两种喂养方法中,一组肝细胞出现一些可逆性损伤,而另一组肝细胞结构与正常组近似。提示 SOD 作为抗氧化剂对食入大剂量氟的大鼠肝细胞有保护作用。     

TNFα/ActD诱导的大鼠肝细胞凋亡超微结构观察

近年来的研究表明 ,在肝细胞损伤过程中细胞凋亡先于细胞坏死 ,而肿瘤坏死因子 (ＴＮＦα)可诱发肝细胞凋亡并参与肝脏疾病的发病过程。据报道 ,体外单独使用ＴＮＦα并不能引起细胞凋亡 ,须与氨基半乳糖 (ＧａＩＮ)或放线菌素Ｄ(ＡｃｔＤ)合用才可显示出明显的毒性作用。本研究即是以透射电镜观察ＴＮＦα诱导ＡｃｔＤ致敏的大鼠肝细胞凋亡的超微结构改变。材料与方法一、实验动物Ｗｉｓｔａｒ雄性大鼠 ,体重 80～ 10 0ｇ ,白求恩国际和平医院实验动物中心提供。二、实验药物ＲＰＭ 1 16 40培养液为Ｇｉｂｃｏ公司产品 ,小牛血清为杭州四季…     

黄绿青霉素对低硒低蛋白大鼠心肌损伤的生化检测

目的 观察黄绿青霉素(CIT)对低硒低蛋白大鼠心肌损伤在生化水平上的特点.方法 48只4周龄Wistar雄性大鼠,2×2析因设计,按体质量随机分为4组:低硒低蛋白加CIT组、低硒低蛋白组、常硒常蛋白加CIT组、常硒常蛋白组,每组12只.分别采用低硒低蛋白和常硒常蛋白饲料喂养大鼠至第12周后,加毒素组大鼠饲料中加入8mg·kg-1·d-1 CIT喂养至第20周,再加量至10 mg·kg-1·d-1 CIT继续喂养至22周,股动脉采血及取心脏,检测血清肌钙蛋白Ⅰ(Tn-Ⅰ)、白蛋白水平,肌酸激酶(CK)、谷胱甘肽过氧化物酶( GSH-Px)以及心肌超氧化物歧化酶(SOD)活性,总抗氧化能力(T-AOC).结果 “硒+蛋白”与CIT对大鼠终来体质量、白蛋白和Tn-Ⅰ存在交互作用(F值分别为8.186、6.160、19.183,P均＜0.05),对12周大鼠体质量,22周血清GSH-Px、CK水平以及心肌SOD、T-AOC活性的影响不存在交互作用(F值分别为1.633、1.987、0.075、0.474、1.145,P均＞ 0.05).在低硒低蛋白背景下,加CIT组血清白蛋白和Tn-Ⅰ水平[(42.88±1.19)g/L,(668.6±55.8) ng/L]均低于无CIT组[(47.59±1.05)g/L,(989.3±49.2)ng/L,P均＜0.05].“硒+蛋白”对12周大鼠体质量、22周血清GSH-Px活性以及心肌SOD、T-AOC有影响(F值分别为96.860、58.086、4.475、25.485,P均＜0.05).低硒低蛋白两组12周大鼠体质量[(186.33±7.89)、(197.83±7.89)g]均低于常硒常蛋白两组[(274.08±7.89)、(265.42±7.89)g,P均＜0.05];低硒低蛋白两组[(317.5±102.6)、(296.9±90.5)U/L]血清GSH-Px活性均低于常硒常蛋白两组[(926.1±110.9)、(1181.7±85.9) U/L,P均＜0.05];低硒低蛋白两组心肌SOD活性[ (65.22±5.91)×106、(62.68±5.61)× 106 U/kg]均低于常硒常蛋白两组[(74.07±7.24)×106、(80.07±5.91)×106 U/kg,P均＜0.05];低硒低蛋白两组心肌T-AOC活性[(1.138±0.086)×106、(0.806±0.081)× 106U/kg]均低于常硒常蛋白两组[(1.688±0.105)×106、(1.163±0.086)×106 U/kg,P均＜0.05].结论 低硒低蛋白模型复制成功,CIT对低硒低蛋白大鼠心肌损伤在生化水平未发现有规律性效应,有待进一步研究确定.     

抗脂肪肝冲剂对大鼠乙醇性肝损伤防治的实验研究

目的：探讨抗脂肪肝冲剂对乙醇性肝损伤大鼠脂质代谢的影响,寻找针对乙醇所致脂肪肝的有效药物,方法：采用乙醇慢性肝损伤模型,观察大鼠肝组织总胆固醇（TC）,及甘油三酯（TG）的含量,结果：抗脂肪肝冲剂降低乙醇性肝损伤大鼠肝组织TC,TG的含量,与模型组比较P＜0．05（小剂量组）,P＜0．01（中,大剂量组）,病理检查,抗脂肪冲剂各剂量组与模型组比较P＜0．01,结论：抗脂肪肝冲剂能够降低乙醇性肝损伤大鼠肝组织TG,TC的含量,具有防治乙醇性脂肪肝的作用。     

Neuroimaging in Alcoholism: Ethanol and Brain Damage

This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The co-chairs were Karl Mann and Ingrid Agartz. The presentations were (1) Neuropathological changes in alcohol-related brain damage, by Clive Harper; (2) Regional brain volumes including the hippocampus and monoamine metabolites in alcohol dependence, by Ingrid Agartz, Susan Shoaf, Robert R, Rawlings, Reza Momenan, and Daniel W Hommer; (3) Diffusion tensor abnormalities in imaging of white matter alcoholism, by Adolf Pfefferbaum and Edith V. Sullivan; (4) Use of functional MRI to evaluate brain activity during alcohol cue exposure in alcoholics: Relationship to craving, by Raymond F. Anton, David J. Drobes, and Mark S. George; and (5) μ-Opiate receptor availability in alcoholism: First results from a positron emission tomography study, by Karl Mann, Roland Bares, Hans-Juergen Machulla, Goetz Mundle, Matthias Reimold, and Andreas Heinz.     

Morphological and biochemical effects of weekend alcohol consumption in rats: Role of concentration and gender

AIM To examine the association between weekend alcohol consumption and the biochemical and histological alterations at two different concentrations of alcohol in both genders in rats.METHODS Wistar rats weighing 170-200 g were divided into groups as follows:(1) Control groups; and(2) weekend alcohol-consumption group: 2 d/weekly per 12 wk, at two different concentrations:(1) Group of males or females with a consumption of a solution of alcohol at 40%; and(2) group of males or females with a consumption of a solution of alcohol at 5%. At the end of the experiment, serum and liver samples were obtained. The following enzymes and metabolites were determined in serum: Alanine Aminotransferase(ALT), Aspartate Aminotransferase(AST), Lactate Dehydrogenase, and Gamma-Glutamyltransferase, and glucose, triglycerides, cholesterol, bilirubin, and albumin. Liver samples from each group were employed to analyze morphological abnormalities by light microscopy.RESULTS In all of the weekend alcohol-consumption groups, AST activity presented a significant, 10-fold rise. Regarding ALT activity, the groups with weekend alcohol consumption presented a significant increase that was six times greater. Bilirubin levels increased significantly in both groups of females. We observed a significant increase in the parameters of fatty change and inflammation due to weekend alcohol consumption. Only the group of females that consumed alcohol at 40% presented slight hepatocel ular disorganization CONCLUSION The results obtained herein provide solid evidence that weekend alcohol consumption gives rise to liver damage, demonstrated by biochemical and histological alterations, first manifested acutely, and prolonged weekend alcohol consumption can cause greater, irreversible damage.     

Pseudoxanthoma Elasticum (PXE): Ultrastructural and Biochemical Study on Proteoglycan and Proteoglycan-Associated Material Produced by Skin Fibroblasts In Vitro

Pseudoxanthoma elasticum is a genetic disease characterized by progressive mineralization of elastic fibers. Previous studies suggested that other components, apart from elastin, might be involved in the alterations of this connective tissue disorder (Martinez-Hernandez and Huf f er, 1974; Pasquali Ronchetti et al., 1981; 1986). Evidence is presented that proteoglycan metabolism is altered in PXE-affected patient. Urinary GAGs suggests an increased degradation of glucosamine-containing GAGs in the patient. Pulse and chase experiments on in vitro skin fibroblasts indicated a decreased rate of synthesis of ³⁵SO₄ containing GAGs or an increase of their turnover rate in PXE. Moreover, when PGs produced from skin fibroblasts were identified by ultracentrifugation and gel filtration in associative conditions, PXE fibroblasts produced a significantly higher amount of the high molecular weight fraction of sulfated PGs. This high molecular weight material was present both in the medium and in the matrix and disappeared under dissociative conditions or after treatment with hyaluronidase or with pancreas elastase. By electron microscopy, PXE fibroblasts appeared to produce and secrete an enormous amount of toluidine blue 0 positive material organized as filaments and amorphous masses. These data are in agreement with previous observations of the presence of abnormal masses of microfilaments, in the dermis of PXE patients, which were sensitive to hyaluronidase and partially to trypsin and elastase (Pasquali Ronchetti et al., 1986). The results seem to confirm that at least some of the alterations of connective tissues in PXE are due to abnormal PGs metabolism and to their tendency to form abnormal aggregates in the extracellular space.     

Protective Effects of Rutin on Mitochondrial Damage in Isoproterenol-Induced Cardiotoxic Rats: An In Vivo and In Vitro Study

Consumption of diets rich in flavonoids is associated with reduced risk of cardiovascular diseases such as myocardial infarction. Cardiotoxicity was induced in rats by subcutaneous injection of isoproterenol at an interval of 24 h for 2 days. Isoproterenol-induced rats showed a significant increase in the levels of heart mitochondrial lipids, lipid peroxidation products, calcium and a significant decrease in the activities/levels of mitochondrial antioxidants, enzymes and adenosine triphosphate. Isoproterenol-induced rats also showed an increase in the intensities of serum lactate dehydrogenase-1 and 2 isoenzyme bands. Pretreatment with rutin at the dose of 80 mg/kg daily for 42 days to isoproterenol-induced rats prevented all the biochemical alterations. Transmission electron microscopic study also confirmed the protective effects of rutin on the structure of heart mitochondria. Thus, rutin reduced the extent of mitochondrial damage induced by isoproterenol and prevented cardiac mitochondrial dysfunction. The possible mechanisms for the observed effects of rutin could be due to scavenging free radicals, lowering lipid peroxides, lipids and calcium, improving multienzyme activities, glutathione levels, adenosine triphosphate levels, thereby improving cardiac mitochondrial structure and function. This study may have a significant impact on myocardial infarcted patients.     

Hepatocellular carcinoma incidence trends in Canada: analysis by birth cohort and period of diagnosis

Background and Aims: We examined birth cohort and calendar period trends in hepatocellular carcinoma (HCC) incidence in Canada (1976–2000). We also projected HCC incidence rates through 2015. Patients and methods: Data were obtained from the Canadian Cancer Registry on all cases of HCC diagnosed among persons aged 20 years and older in Canada from 1976 to 2000 and was used to describe trends in HCC incidence rates. Results: We found that age‐adjusted HCC incidence rates increased faster in males compared with females, 3.4% per year [95% confidence interval (CI): 3.0–3.8%] vs 2.2% per year (95% CI: 1.5–2.8%). An increasing birth cohort trend accelerated among males around the 1940 birth cohort and decelerated among females around the 1935 birth cohort. For calendar period trends, the increasing HCC risk was relatively constant over time among males whereas there was an acceleration in HCC risk around 1988 among females. Age‐adjusted HCC incidence rates were projected to increase 73% in males and 28% in females from 1996 to 2015. Conclusions: Our results suggest that HCC incidence rates will continue to increase in Canada during the next decade as persons born in more recent birth cohorts, who face a relatively greater risk for HCC, age.     

Collaborative Research: An Effective Way to Collect Data for Stock Assessments and Evaluate Marine Protected Areas in California

Abstract

Collaborative fisheries research (in contrast to cooperative research) is based on the intellectual partnership between scientists and fishermen and is an effective way to collect data for stock assessments and to evaluate marine protected areas. Collaborative fisheries research is discussed in the context of co-management of marine resources and how it contributes to a more democratic form of fisheries management. Many benefits result from working together, including (1) the incorporation of fishers' knowledge and expertise into the management process and (2) the development of shared perspectives derived through science-based investigations on the status of marine resources. The California Collaborative Fisheries Research Program was formed in 2006 to participate in the monitoring of marine reserves established through California's Marine Life Protection Act. This program has shown that it can serve as a model for other areas that are trying to implement collaborative research and that collaborative research can greatly contribute to the realization of community-based co-management of marine resources.     

Protective Effects of Aqueous Garlic Extract in Reducing Water Avoidance Stress-Induced Degeneration of the Stomach, Ileum, and Liver: Morphological and Biochemical Study

We investigated the effect of aqueous garlic extract (AGE) on water avoidance stress (WAS)-induced degeneration of the gastric and ileal mucosa and liver parenchyma. Wistar albino rats were exposed to WAS (WAS group) for 5 days. After exposure of the animals to WAS, a 1 ml/kg aqueous garlic extract (AGE) was injected i.p. (WAS+AGE group). The stomach, ileum, and liver samples were investigated under light microscope for general morphology. Topography of gastric and ileal mucosa was investigated by scanning electron microscopy, and hepatocyte ultastructure by transmission electron micsroscopy. Malondialdehyde (MDA) and glutathione (GSH) levels of all tissues were also determined. In the WAS group, the epithelium of the stomach showed ulceration in some areas, dilatations of the gastric glands, and degeneration of gastric glandular cells. Severe vascular congestion and degeneration of ileal epithelium were observed. Prominent vascular congestion and dilated sinusoids, activated Kupffer cells with prominent morphology, dilated granular endoplasmic reticulum membranes, and focal picnotic nuclei were observed in liver parenchyma. AGE treatment reduced the degeneration of the gastric and ileal mucosa and liver parenchyma. Increased MDA levels and decreased GSH levels in the WAS group were reversed to control values after AGE treatment. Based on these results, AGE treatment significantly prevented WAS-induced degeneration in both morphology and biochemistry of gastrointestinal mucosa and liver parenchyma due to its potent free radical scavenging and antioxidant properties.