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Laser capture microdissection and genetic analysis of carbon-labeled Kupffer cells

World Journal of Gastroenterology 2009年第14期15卷 1708-1718页

作者： stephan gehring Edmond Sabo Maryann E San Martin Elizabeth M Dickson Chao-Wen Cheng Stephen H Gregory Department of Medicine Rhode Island Hospital and The Warren Alpert Medical School of Brown University Providence RI 02903 United States Department of Pathology Rhode Island Hospital and The Warren Alpert Medical School of Brown University Providence RI 02903 United States zentrum fur Kinder- und Jugendmedizin der Universitaitsmedizin der Johannes Gutenberg-Universitait 55101 Mainz Germany

AIM: To develop a method of labeling and microdissecting mouse Kupffer cells within an extraordinarily short period of time using laser capture microdissection (LCM). METHODS: Tissues are complex structures comprised of a heterogeneous population ofinterconnected cells. LCM offers a method ofisolating a single cell type from specific regions of a tissue section. LCM is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCM prior to mRNA isolation can be elaborate, thereby subjecting the RNA to considerable degradation. Kupffer cell labeling is achieved byinjecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model. RESULTS: mRNA extracted from the microdissected cell population displayed marked increases in colonystimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. Gene expression by Kupffer cells derived from bile-duct-ligated, versus sham-operated, mice was compared. Microarray analysis revealed a significant (2.5-fold, q value < 10) change in 493 genes. Based on this fold-change and a standardized PubMed search, 10 genes were identified that were relevant to the ability of Kupffer cells to suppress liver injury. CONCLUSION: The methodology outlined herein provides an approach to isolating high quality RNA from Kupffer cells, without altering the tissue integrity.

关键词： Kupffer细胞激光捕获显微切割遗传分析液晶显示模块肝损伤模型组织切片 mRNA 基因表达

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Prevalence of hepatitis B virus precore stop codon mutations in chronically infected children

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World Journal of Gastroenterology 2006年第14期12卷 2235-2238页

作者： Philip Wintermeyer Patrick Gerner stephan gehring Afshin Karimi Stefan Wirth Children's Hospital Helios Klinikum Wuppertal Witten-Herdecke UniversityGermany Children's Hospital University of Mainz Germany

瞄准：为了发现是否在 precore 站的流行有有效差量，在在积极的 HBeAg 和 anti-HBe 之间的变化上哄骗积极孩子。方法：我们调查了 155 个欧洲孩子的一张大小儿科的人口(意味着年龄 10.9 年) 与由 PCR 并且直接定序的长期的肝炎 B。90 ... 详细信息

瞄准：为了发现是否在 precore 站的流行有有效差量，在在积极的 HBeAg 和 anti-HBe 之间的变化上哄骗积极孩子。方法：我们调查了 155 个欧洲孩子的一张大小儿科的人口(意味着年龄 10.9 年) 与由 PCR 并且直接定序的长期的肝炎 B。90 是积极的 HBeAg 和 65 有的 seroconversion 到 anti-HBe。另外， genotyping 被执行。结果：(48%) 74 定序的 HBV 紧张被归因于遗传型 D 并且 81 (52%) 到遗传型 A。在 90 个 HBeAg 积极病人的组， 2 (2.2%) 变化上的 1896-G-to-A 转变导致 precore 站鳕鱼被发现，并且在 65 anti-HBe 的组积极孩子，(7.7%) 5 被识别 harbouring HBeAg 减异种。差别不是统计上重要的(P = 0.13 ) 。结论：减占优势的病毒的 HB 紧张在欧洲孩子在长期的肝炎 B 期间起一个次要的作用的变体的 HBeAg。

关键词：乙型病毒肝炎基因突变抗原密码子

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Prevalence of autoantibodies and the risk of autoimmune thyroid disease in children with chronic hepatitis C virus infection treated with interferon-α

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World Journal of Gastroenterology 2006年第36期12卷 5787-5792页

作者： stephan gehring Ulrike Kullmer Sabine Koeppelmann Patrick Gerner Philip Wintermeyer Stefan Wirth The Liver Research Center Department of Medicine Rhode Island Hospital and Brown Medical School Providence Rhode Island USA and Kinderklinik der Johannes-Gutenberg-Universitt Mainz Germany Kinderklinik der Johannes-Gutenberg-Universitt Mainz Germany Zentrum für Kinder-und Jugendmedizin HELIOS Klinikum Wuppertal Witten-Herdecke-University Germany

AIM: To evaluate the prevalence of autoantibodies in chronic hepatitis C virus (HCV)-infected children focusing on thyroid autoimmunity. METHODS: We investigated the prevalence of auto- antibodies in 123 chronic HCV-infected children before, during and after monotherapy with interferon-alpha (IFN-α) or combined treatment with interferon-α or peginterferon-α and ribavirin. Besides antibodies against smooth muscle (SMA), nuclei (ANA), and liver/kidney microsomes (LKM), the incidence of anti- thyroid peroxidase antibodies as well as thyroid function parameters (TSH, FT3 and FT4) were determined. RESULTS: We found that 8% of children had autoantibodies before treatment. During treatment, 18% of children were found positive for at least one autoantibody; 15.5% of children developed pathologic thyroid values during IFN-α treatment compared to only one child before therapy. Six children had to be substituted while developing laboratory signs of hypothyroidism. CONCLUSION: Our data indicate a strong correlation between interferon-α treatment and autoimmune phenomena, notably the emergence of thyroid antibodies. The fact that some children required hormone replacement underlines the need of close monitoring in particularly those who respond to therapy and have to be treated for more than 6 mo.

关键词：自身抗体甲状腺疾病自身免疫慢性丙性肝炎

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