Increased expression of chondroitin sulphate proteoglycans in rat hepatocellular carcinoma tissues

AIM: To investigate the expression of chondroitin sulphate proteoglycans (CSPGs) in rat liver tissues of hepatocellular carcinoma (HCC).METHODS: Thirty male Sprague Dawley rats were randomly divided into two groups: control group (n = 10) and HCC model group (n = 20). Rats in the HCC model groups were intragastrically administrated with 0.2% (w/v) N-diethylnitrosamine (DEN) every 5 d for 16 wk, whereas 0.9% (w/v) normal saline was administered to rats in the control group. After 16 wk from the initiation of experiment, all rats were killed and livers were collected and fixed in 4% (w/v) paraformaldehyde. All tissues were embedded in paraffin and sectioned. Histological staining (hematoxylin and eosin and Toluidine blue) was performed to demonstrate the onset of HCC and the content of sulphated glycosaminoglycan (sGAG). Immunohistochemical staining was performed to investigate the expression of chondroitin sulphate (CS)/dermatan sulphate (DS)-GAG, heparan sulphate (HS)-GAG, keratan sulphate (KS)-GAG in liver tissues. Furthermore, expression and distribution of CSPG family members, including aggrecan, versican, biglycan and decorin in liver tissues, were also immunohistochemically determined.RESULTS: After 16 wk administration of DEN, malignant nodules were observed on the surface of livers from the HCC model group, and their hepatic lobule structures appeared largely disrupted under microscope. Toluidine blue staining demonstrated that there was an significant increase in sGAG content in HCC tissues when compared with that in the normal liver tissues from the control group [0.37 ± 0.05 integrated optical density per stained area (IOD/area) and 0.21 ± 0.01 IOD/area, P < 0.05]. Immunohistochemical studies demonstrated that this increased sGAG in HCC tissues was induced by an elevated expression of CS/DS (0.28 ± 0.02 IOD/area and 0.18 ± 0.02 IOD/area, P < 0.05) and HS (0.30 ± 0.03 IOD/area and 0.17 ± 0.02 IOD/area, P < 0.01) but not KS GAGs in HCC tissues. Further studies thereby were performed to investigate the expression and distribution of several CSPG components in HCC tissues, including aggrecan, versican, biglycan and decorin. Interestingly, there was a distinct distribution pattern for these CSPG components between HCC tissues and the normal tissues. Positive staining of aggrecan, biglycan and decorin was localized in hepatic membrane and/or pericellular matrix in normal liver tissues; however, their expression was mainly observed in the cytoplasm, cell membranes in hepatoma cells and/or pericellular matrix within HCC tissues. Semi-quantitative analysis indicated that there was a higher level of expression of aggrecan (0.43 ± 0.01 and 0.35 ± 0.03, P < 0.05), biglycan (0.32 ± 0.01 and 0.25 ± 0.01, P < 0.001) and decorin (0.29 ± 0.01 and 0.26 ± 0.01, P < 0.05) in HCC tissues compared with that in the normal liver tissues. Very weak versican positive staining was observed in hepatocytes near central vein in normal liver tissues; however there was an intensive versican distribution in fibrosis septa between the hepatoma nodules. Semi-quantitative analysis indicated that the positive rate of versican in hepatoma tissues from the HCC model group was much higher than that in the control group (33.61% and 21.28%, P < 0.05). There was no positive staining in lumican and keratocan, two major KSPGs, in either normal or HCC liver tissues.CONCLUSION: CSPGs play important roles in the onset and progression of HCC, and may provide potential therapeutic targets and clinical biomarkers for this prevalent tumor in humans.     

不同急性胰腺炎凝血功能紊乱的临床研究

目的 探讨急性胰腺炎时凝血功能的变化.方法 选择急性胰腺炎患者55例,其中仅有局部表现的患者20例(局部症状组),有全身炎症反应综合征(SIRS)而无器官功能障碍的患者20例(SIRS组),出现器官功能障碍的患者10例(器官功能障碍组),死亡患者5例(死亡组).择期胆囊切除术术前检查凝血功能正常的同期住院患者10例(对照组).检测上述患者的血小板计数、凝血酶原时间、活化部分凝血酶时间和纤维蛋白原.结果 局部症状组血小板计数、凝血酶原时间、活化部分凝血酶时间和纤维蛋白原与对照组比较差异均无统计学意义;SIRS组、器官功能障碍组和死亡组血小板计数显著低于对照组,凝血酶原时间、活化部分凝血酶时间显著长于对照组,纤维蛋白原显著高于对照组.结论 急性胰腺炎发展到SIRS、器官功能障碍和死亡阶段时,凝血功能表现为紊乱状态.     

脑尔康对AD模型小鼠脑内APP,Aβ表达的影响

目的:观察复方中药脑尔康对慢性铝中毒阿尔茨海默病(AD)模型小鼠学习记忆及脑内淀粉样前体蛋白(APP),β淀粉样蛋白(Aβ)表达的影响,探讨该药抗痴呆的分子机制。方法:60只昆明小鼠随机分为空白对照、模型、西药吡啦西坦、脑尔康高、中、低剂量6组,除空白组外,其余各组均采用氯化铝(AlCl3)溶液以100 mg.kg-1ip造模,空白组给予等量生理盐水ip,3个剂量脑尔康组给予脑尔康(34,17,8.5 g.kg-1.d-1)连续ig 50 d,吡啦西坦组给予吡啦西坦(10 g.L-1)ig,空白组和模型组给予等量生理盐水ig。采用跳台法检测AD模型小鼠学习记忆成绩,用免疫组织化学方法观察脑尔康对AD模型小鼠脑皮层及海马结构区域APP,Aβ表达的影响,对APP,Aβ阳性反应物质采用图像信号采集与分析系统进行灰度分析。结果:与对照组比较,模型组记忆潜伏期明显缩短(P<0.01),学习潜伏期明显延长(P<0.05),学习与记忆的错误次数明显增多(P<0.01);用药各组分别与模型组比较记忆潜伏期明显延长(P<0.01),学习潜伏期明显缩短(P<0.05),学习与记忆的错误次数明显减少(P<0.01)。各用药组模型小鼠脑内APP,Aβ的表达均不同程度减少(P<0.05或P<0.01);脑尔康3个剂量组比较存在明显量效关系。结论:脑尔康对慢性铝中毒AD模型具有显著的抗痴呆作用,其作用机制可能与调控APP,Aβ表达有关。     

Construction of targeted plasmid vector pcDNA3.1-Egr.1p-p16 and its expression in pancreatic cancer JF305 cells induced by radiation in vitro

To construct pcDNA3.1-Egr.1p-p16 recombinant plasmid and investigate the expression of p16 in pancreatic cancer JF305 cells induced by radiation and the feasibility of gene radiotherapy for pancreatic carcinoma.     

胸腰椎骨折分型与临床治疗方法探讨

<正>胸腰椎骨折(Thoracolubar fractures)指发生在所有胸椎、腰椎的脊柱骨折。胸腰段骨折(Fractures of thoracolumbar junction)特指发生在T11-L2段的脊柱骨折。在脊柱骨折中,其发病率约为50%[1-2]。胸腰段骨折是脊柱骨折中最常见的部位,因此,     

The PPARγ agonist rosiglitazone prevents cognitive impairment by inhibiting astrocyte activation and oxidative stress following pilocarpine-induced status epilepticus

Epilepsy is commonly associated with cognitive impairment. Astrocyte activation and oxidative stress occur following seizures, and play a role in the pathological injury of epilepsy with cognitive impairment. The peroxisome proliferator-activated receptor gamma (PPARγ) has been shown to exhibit neuroprotective and antioxidative effects in CNS diseases. Thus, we hypothesized that rosiglitazone, a PPARγ agonist, would prevent cognitive impairment by inhibiting astrocyte activation and regulating glutathione (GSH) homeostasis after status epilepticus (SE). Using a lithium pilocarpine-induced SE model, we found that rosiglitazone significantly prevented cognitive impairment induced by SE, and potently inhibited astrocyte activation with maintenance of GSH homeostasis in the hippocampus after SE. These protective effects were significantly reversed by co-treatment with the PPARγ antagonist T0070907. These data suggest that rosiglitazone can improve cognitive impairment, and inhibit astrocyte activation and oxidative damage following SE. Rosiglitazone may be a promising agent for treatment of epilepsy involving SE-induced cognitive impairment.     

小剂量ATP静脉注射终止阵发性室上性心动过速30例发作疗效观察

目的:探讨小剂量三磷酸腺苷(ATP)快速静脉注射终止阵发性室上性心动过速(PSVT)疗效。方法:对30例PSVT快速静脉注射小剂量ATP,并观察疗效。结果:30例病人中27例效果良好,有效率为90%。结论:小剂量ATP快速静脉注射终止PSVT,作用快,转复效果良好。     

小分子化合物S1抑制K562细胞增殖及其机理研究

目的:探讨小分子化合物S1对K562细胞的抑制作用及机理,为S1治疗慢性粒细胞白血病（chronic myeloid leukemia,CML）提供理论与实验基础。方法:MTT法检测不同浓度S1对K562细胞增殖的抑制作用;应用Chou-Talalay计算联合指数,观察单独应用S1与S1联合阿糖胞苷对K562细胞增殖抑制的协同作用;Western-blotting方法检测不同浓度S1作用后K562细胞系中Bcl-2蛋白表达水平的变化。结果:与对照组相比,S1可以抑制K562细胞的增殖（P<0.05）;S1与阿糖胞苷联合应用对K562细胞的生长抑制具有协同作用,联合指数CI<1;S1可下调K562细胞系中抗凋亡蛋白Bcl-2的表达。结论:小分子化合物可通过下调抗凋亡蛋白Bcl-2的表达抑制K652细胞的增殖,并与阿糖胞苷具有协同作用。     

Acute stimulation of glucagon secretion by linoleic acid results from GPR40 activation and [Ca2+]i increase in pancreatic islet {alpha}-cells

The role of free fatty acids (FFAs) in glucagon secretion has not been well established, and the involvement of FFA receptor GPR40 and its downstream signaling pathways in regulating glucagon secretion are rarely demonstrated. In this study, it was found that linoleic acid (LA) acutely stimulated glucagon secretion from primary cultured rat pancreatic islets. LA at 20 and 40?μmol/l dose-dependently increased glucagon secretion both at 3?mmol/l glucose and at 15?mmol/l glucose, although 15?mmol/l glucose reduced basal glucagon levels. LA induced an increase in cytoplasmic free calcium concentrations ([Ca(2)(+)](i)) in identified rat α-cells, which is reflected by increased Fluo-3 intensity under confocal microscopy recording. The increase in [Ca(2)(+)](i) was partly inhibited by removal of extracellular Ca(2)(+) and eliminated overall by further exhaustion of intracellular Ca(2)(+) stores using thapsigargin treatment, suggesting that both Ca(2)(+) release and Ca(2)(+) influx contributed to the LA-stimulated increase in [Ca(2)(+)](i) in α-cells. Double immunocytochemical stainings showed that GPR40 was expressed in glucagon-positive α-cells. LA-stimulated increase in [Ca(2)(+)](i) was blocked by inhibition of GPR40 expression in α-cells after GPR40-specific antisense treatment. The inhibition of phospholipase C activity by U73122 also blocked the increase in [Ca(2)(+)](i) by LA. It is concluded that LA activates GPR40 and phospholipase C (and downstream signaling pathways) to increase Ca(2)(+) release and associated Ca(2)(+) influx through Ca(2)(+) channels, resulting in increase in [Ca(2)(+)](i) and glucagon secretion.