Endoscopic stenting of the gallbladder for symptomatic gallbladder disease in patients with end-stage liver disease awaiting orthotopic liver transplantation.

Cholecystectomy in patients with advanced cirrhosis is associated with excessive morbidity and mortality. Because open cholecystectomy in patients with Child's class C cirrhosis has a reported mortality rate as high as 83%, symptomatic gallbladder disease in patients awaiting orthotopic liver transplantation (OLT) poses a unique clinical problem. The goal of this study is to determine whether the treatment of symptomatic gallbladder disease with endoscopic stenting of the gallbladder effectively reduces biliary symptoms and complications or the need for cholecystectomy. Thirteen patients with symptomatic gallbladder disease with and without cholelithiasis and advanced cirrhosis who were candidates for OLT underwent placement of a biliary stent from the gallbladder to the duodenum at endoscopic retrograde cholangiography. In each patient, biliary symptoms and complications ceased after stent placement. Seven patients underwent successful OLT 1 to 24 months after the procedure. One patient subsequently became a noncandidate for OLT and died of diabetes complications 3 years after the procedure. Five others are awaiting OLT (6 to 28 months postprocedure). One patient had recurrent pericholecystic fluid collection requiring percutaneous drainage and antibiotic therapy 8 months after the procedure. No patient has had recurrent symptoms, and currently all patients are free of complications. None required surgical intervention of the gallbladder or biliary tree. We conclude that endoscopic stenting of the gallbladder is the preferred treatment for symptomatic gallbladder disease in patients with end-stage liver disease awaiting OLT. This approach is noninvasive, safe, and effective in preventing potential morbidity and mortality.     

Cytochemical,immunofluorescence, and ultrastructural investigations on polypeptide hormone localization in the islet parenchyma and bile duct mucosa of a cyclostome,Myxine glutinosa

Cells in the islets and bile duct epithelium of the hagfish, Myxine glutinosa, can take up and decarboxylate precursors of biogenic amines. The end product of the process is demonstrated by the freeze-drying and formaldehyde vapor fixation technique. The same cells can be stained by an immunofluorescence reaction for insulin, using guinea pig antiserum to human insulin. Thus, the insulin-producing B-cells of the hagfish are members of the socalled APUD series of polypeptide hormone-producing cells described in mammals.Insulin-containing cells were also observed, forming buds from the bile duct mucosa. Consequently, the islets have obviously been formed from endocrine precursor cells in the bile duct epithelium. Immature extrainsular B-cells of this kind obviously attain APUD characteristics before they show actual insulin production.The content of the characteristic islet cavities was nonimmunoreactive to either anti-human insulin or anti-human C-peptide, and did not take up precursors of biogenic amines. No evidence was obtained of B-cell granule release to the islet cavities. Instead, the secretion granules seemed to be released by emiocytosis to the surrounding connective tissue by secretion of “membrane-release” pattern. Nerve fibers were observed in the connective tissue stroma between the islet lobules but there was no close connection between nerve fibers and B-cells.A pilot study revealed the presence of glucagon-immunoreactive cells in the intestinal epithelium but no cells of this kind were observed in the islet organ or in the bile duct mucosa. No cells in the bile duct mucosa or in the islet parenchyma showed immunoreactivity to gastrin, secretin, or caerulein.     

Inhibition of mammal Na+K+‐ATPase by diterpenes extracted from the Brazilian brown alga Dictyota cervicornis

The dolastane diterpenes 4‐acetoxy‐9,14‐dihydroxydolast‐1(15),7‐diene (1) and 4,7‐diacetoxy‐14‐hydroxydolast‐1(15),8‐diene (2) were isolated from specimens of the alga Dictyota cervicornis collected from the Rio de Janeiro coast, Brazil. Chemical structures of the diterpenes were assigned by 1D and 2D NMR spectral data for the first time. Both substances inhibited Na⁺K⁺‐ATPase preparations from guinea‐pig brain or kidney, with the same inhibitory potency towards enzyme isoforms. The maximal inhibition obtained for 1 was 40% at a concentration of 0.5 mm in the incubation mixture, while it reached 80% for compound 2 at this concentration. Ouabain insensitive ATPases were inhibited by 1, but not by 2. Data comparing the inhibitory potency of these compounds with that of ouabain and oleic acid suggest a higher degree of selectivity of 2 towards the Na⁺K⁺‐pump. Cardiac glycosides such as ouabain are used classically in the treatment of heart failure, but alterations of Na⁺K⁺‐pump activity are also involved in several other diseases. Therefore, the study of compounds interfering with this pump activity is gaining further importance. Copyright © 2009 John Wiley & Sons, Ltd.     

巢式逆转录-聚合酶链反应扩增人骨形成蛋白2全长cDNA及其真核表达载体的构建

目的:利用巢式逆转录-聚合酶链反应扩增的方法,从肌肉组织中扩增人骨形成蛋白2全长cDNA并构建真核表达载体系统。方法:实验于2003-10/2005-10在苏州大学基因工程教研室和北京大学第三医院骨科实验室完成。提取成人肌肉组织内的总RNA,设计内外两对引物以巢式逆转录-聚合酶链反应扩增方法分两次扩增出人骨形成蛋白2全长1188bp基因,经T-A克隆装入pUCM-T质粒载体内,测序验证后,将克隆质粒以Hind Ⅲ和Xba Ⅰ双酶切后与pcDNA3.0载体相连接,构建真核表达载体系统。结果:利用巢式逆转录-聚合酶链反应扩增方法能从成人肌肉组织内扩增出1188bp的人骨形成蛋白2全长cDNA基因,其测序结果显示与Genebank报道序列完全相符。将扩增序列双酶切后与pcDNA3.0载体相连接,经电泳验证,能构建人骨形成蛋白2全长基因的真核表达系统。结论:巢式逆转录-聚合酶链反应扩增方法能从成人肌肉组织内扩增出人骨形成蛋白2全长cDNA基因,并克隆构建真核表达载体系统,为下一步基因组织工程人工骨实验奠定基础。