血管紧张素Ⅱ对心脏成纤维细胞TRPM7通道功能的影响

目的:研究心脏成纤维细胞(CFs)膜上的TRPM7通道的电生理特性。方法:将分离培养出来的乳鼠CFs用6种不同浓度的血管紧张素Ⅱ(AngⅡ)干预至12h,分别测量各组胶原蛋白的表达水平,从而挑选出最佳诱导纤维化的AngⅡ浓度,最佳浓度的AngⅡ分别将CFs干预至5个不同的时间,记录相应的CFs膜上的TRPM7电流及各组的胶原蛋白水平。结果:诱导心脏纤维化最大的AngⅡ浓度是1nmol/L,最佳诱导纤维化浓度干预0,6,12,24,48h后CFs的TRPM7与胶原蛋白Ⅲ的表达水平最初升高,于12h达到最高水平,之后呈现递减水平,且在CFs膜上记录到的TRPM7内外向电流也呈现相应的先增加后降低趋势,与两种蛋白的表达水平正相关。结论:CFs膜上的TRPM7电流在AngⅡ的作用下,呈现先增加后减少的趋势,是AngⅡ介导CFs增殖与凋亡从而导致心脏纤维化的重要途径之一。     

Risk of hepatocellular carcinoma after hepatitis C virus cure

Hepatitis C virus(HCV)is a significant cause of hepatocellular carcinoma(HCC).The direct-acting antivirals marked a new era of HCV therapy and are associated with greater than 95%cure rate.Successful treatment of chronic hepatitis C greatly reduces the risk of HCC.A proportion of patients,especially those with pre-existing cirrhosis,remain at risk for HCC despite sustained virologic response(SVR).Diabetes mellitus,hepatic steatosis,alcohol consumption and lack of fibrosis regression are associated with risks of HCC after HCV cure.Noninvasive modalities such as aspartate aminotransferase to platelet ratio index and fibrosis-4 index and transient elastography have been used to monitor hepatic fibrosis.More recently,various fibrosis scores have been combined with clinical parameters and other novel biomarkers to predict risks of HCC for patients who achieved SVR.These models still need to be validated and standardized prior to applying to routine clinical care.     

慢性乙型肝炎血清病毒载量水平与肝硬化的关系

目的 探讨慢性乙型肝炎患者病程中血清乙型肝炎病毒(HBV)-DNA水平的变化与肝硬化发生的关系.方法 收集2001至2007年经肝穿刺确诊的239例慢性乙型肝炎患者.中位随访时间28个月,检测入选和随访终点的血清HBV-DNA水平,观察肝硬化发生情况.结果 发生肝硬化者较无肝硬化者年龄更大,随访终点HBV-DNA水平更高,但两组入选时HBV-DNA水平差异无统计学意义(P=0.531).Kaplan-Meier法生存分析显示,随访终点HBV-DNA水平越高,发生肝硬化比例亦越高(X2=11.736,P=0.019).Cox比例风险模型显示,随访终点HBV-DNA水平、入选时的肝组织纤维化分期、乙型肝炎病毒e抗原阴性和γ-谷氨酰转肽酶水平为预示肝硬化发生的危险因素,风险比分别为1.898、1.918、8.976、1.006.结论 随访终点HBV-DNA和慢性乙型肝炎肝硬化的发生密切相关.     

一氧化氮对感染日本血吸虫小鼠肝脏纤维化程度的影响

目的　研究一氧化氮 (NO)对感染日本血吸虫小鼠早期肝脏纤维化的影响。方法　 NMRI小鼠感染日本血吸虫后第 2 3天起 ,连续用诱导型一氧化氮合酶 (i NOS)的抑制剂氨基胍(Am inoguanidine,AMG)处理小鼠 ,同时设未经抑制的对照组 ,分别在感染后第 38、4 5天观察小鼠肝脏的病理变化。天狼猩红染色 -偏振光显微镜观察并结合图像分析的方法 ,分析肝脏中 、 型胶原的动态变化 ;化学显色法检测肝脏匀浆中能间接反映肝脏纤维化程度的羟脯氨酸的浓度。结果　感染后 38d,肝脏中 、 型胶原的增生程度及 、 型胶原面积比值明显高于对照组 ,但羟脯氨酸的含量与对照组相比差异无显著性 (P>0 .0 5 ) ;感染后 4 5 d,肝脏中 型胶原的增生程度和 、 型胶原面积比值明显高于对照组 ,AMG处理组中羟脯氨酸的含量较对照组高 (P<0 .0 5 )。结论　 NO可抑制日本血吸虫感染小鼠肝脏早期纤维化     

汉防己甲素阻抑大鼠肝纤维化的实验研究

目的：研究汉防己甲素对实验性肝纤维化的作用。方法：用CCl4制备大鼠肝纤维化模型，以鳖甲作为对照。观察汉防己甲素对大鼠肝纤维化各项指标的影响。结果：组织学和生化分析表明，汉防己甲素能减少胶原在肝内沉积；免疫荧光半定量分析提示汉防己甲素组肝内Ⅰ型胶原含量较对照组显著减少；电镜观察发现汉防己甲素组大鼠肝内有些纤维母细胞胞浆中出现髓鞘样结构及自噬体。结论：汉防己甲素具有阻抑肝纤维化的作用，其作用机制可能与该药对纤维母细胞的致细胞病变作用有关。     

干扰素α干预对大鼠胰腺纤维化的影响

目的 观察干扰素α(INF-α)对DDC诱导的胰腺纤维化大鼠模型纤维增生程度、胰腺星状细胞活化标志物α-平滑肌肌动蛋白(α-SMA)和细胞外基质成分Ⅲ型胶原蛋白表达的影响.方法 Wistar大鼠40只随机数字法分成对照组、纤维化组和干扰素组.纤维化组和干扰素组每周2次腹腔内注射DDC,干扰素组在造模同时每天皮下注射INF-α10万U.6周末取材,光镜下观察胰腺病理学变化.免疫组化法测定胰腺组织中α-SMA、Ⅲ型胶原蛋白的表达.结果 第4周起纤维化组大鼠体重增长缓慢甚至下降,干扰素组体重仍缓慢增长,5周后两组差异显著[(309.8±19.7)g与(277.3±19.9)g,P＜0.05].纤维化组胰腺组织纤维化表现明显,纤维化分值、Masson染色值、α-SMA和Ⅲ型胶原蛋白相对表达量分别为2.679±0.899、218.713±36.102、148.971±30.686和88.142±42.581;干扰素组纤维化减轻,上述指标分别为1.952±0.219、114.732±24.912、77.237±9.275和59.952±25.498,均较纤维化组显著降低(P＜0.05).结论 给予INF-α能显著减轻纤维化程度和α-SMA、Ⅲ型胶原表达,对DDC诱导的大鼠胰腺纤维化有一定的预防作用.     

Immunohistochemical detection of arginase-I expression in formalin-fixed lung and other tissues

Abstract

Arginases are a family of enzymes that convert l-arginine to l-ornithine and urea. Alterations in expression of the isoform arginase-I are increasingly recognized in lung diseases such as asthma and cystic fibrosis. To define expression of murine arginase-I in formalin-fixed tissues, including lung, an immunohistochemical protocol was validated in murine liver, a tissue that has distinct zonal arginase-I expression, making it a useful control. In the lung, arginase-I immunostaining was observed in airway surface epithelium and this decreased from large to small airways, with a preferential staining of ciliated epithelium versus Clara cells and alveolar epithelia. In submucosal glands, the ducts and serous acini had moderate immunostaining, which was absent in mucous cells. Focal immunostaining was observed in alveolar macrophages, endothelial cells, pulmonary vein cardiomyocytes, pulmonary artery smooth muscle, airway smooth muscle, and neurons of ganglia of the lung. Arginase-I immunostaining was also detected in other tissues including salivary glands, pancreas, liver, skin, and intestine. Differential immunostaining was observed between sexes in submandibular salivary glands; arginase-I was diffusely expressed in the convoluted granular duct cells of females, but was rarely noted in males. Strain specific differences were not detected. In a mouse with an incidental case of lymphoma, neoplastic lymphocytes lacked arginase-I immunostaining, in contrast to immunostaining detected in non-neoplastic lymphocytes of lymphoid tissues. The use of liver tissue to validate arginase-I immunohistochemistry produced consistent expression patterns in mice, and this approach can be useful to enhance consistency of arginase-I immunohistochemical studies.     

Synthesis and Evaluation of Long-Acting D-Penicillamine Derivatives

This study extends the use of two lathyrogens, β-aminopropionitrile (BAPN) and D-penicillamine (DPA) from daily systemic or local-topical administration to long-time acting agents. This was achieved by converting the hydrophilic drugs into lipophilic derivatives. The synthesis of functional derivatives of DPA consisted in esterification with methyl-, hexyl-, or benzyl alcohols in the presence of thionylchloride. The esters formed were hydrochlorides, acidic and soluble in water. During neutralization in vitro or in vivo by tissue fluid, an oily substance is formed that elutes from a hydrogel polymer at a much slower rate than hydroplilic DPA itself. The degree of lipophilicity, measured as a partition coefficient between octanol/water, was highest for hexyl ester and lowest for methyl ester DPA. A single injection of either DPA hexyl ester HCl or 3-hexyl(amino) propionitrile into the full thickness skin incision wound in rats significantly lowered the breaking strength of the wound 12 days after injection, indicating the interference with collagen cross-linking. Both agents injected into the breast adenocarcinoma in Fisher rats significantly inhibited tumor growth without any signs of local or systemic toxicity. We conclude that these lipophilic lathyrogens with prolonged effectiveness are suitable in the treatment of pathologies, consisting of excessively cross-linked or deposited collagen (fibrotic adhesions, strictures, stenosis, and scar contractures) and in the treatment of single, solitary tumors, malignant and benign.