Analysis of microdissected prostate tissue with ProteinChip arrays--a way to new insights into carcinogenesis and to diagnostic tools

Prostate carcinomas are one of the most common malignancies in western societies. The pathogenesis of this tumor is still poorly understood. These tumors present with two characteristic features: epithelial-mesenchymal interactions, which play a pivotal role for tumor development and most of clinically manifest cancers arise in prostate proper compared to a minority of tumors which develop in the transitional zone. Deciphering the epithelial-mesenchymal cross talk and identification of molecular pecularities of the sub-populations of cells in different zones can therefore help understanding carcinogenesis and development of new, non-invasive tools for the diagnosis and prognosis of prostate carcinomas which has remained a challenge until today. A ProteinChip array technology (SELDI = surface enhanced laser desorption ionization) has been developed recently by Ciphergen Biosystems enabling analysis and profiling of complex protein mixtures from a few cells. This study describes the analysis of approximately 500-1000 freshly obtained prostate cells by SELDI-TOF-MS (surface enhanced laser desorption ionization time-of-flight mass spectrometry). Pure cell populations of stroma, epithelium and tumor cells were selected by laser assisted microdissection. Multiple specific protein patterns were reproducibly detected in the range from 1.5 to 30 kDa in 28 sub-populations of 4 tumorous prostates and 1 control. A specific 4.3 kDa peak was increased in the prostate tumor stroma compared to normal prostate proper and transitional zone stroma and increased in prostate tumor glands compared to normal prostate proper and transitional zone glands. Coupling laser assisted microdissection with SELDI provides tremendous opportunities to identify cell and tumor specific proteins to understand molecular events underlying prostate carcinoma development. It underlines the vast potential of this technology to better understand pathogenesis and identify potential candidates for new specific biomarkers in general which could help to screen for and distinguish disease entities, i.e. between clinically significant and insignificant carcinomas of the prostate.     

The acetylcholine release enhancer linopirdine induces Fos in neocortex of aged rats

Centrally acting cholinergic agents induce the immediate early gene c-fos in the rat brain resulting in transient increases of Fos protein, most notably in the cerebral cortex. In this study we have monitored by Fos immunohistochemistry the effect of the acetylcholine release enhancer linopirdine (DUP996) on the immediate early gene c-fos in brains of 3 months and 30 months old rats. In young rats linopirdine had only a marginal effect on Fos expression. In contrast, in aged rats linopirdine caused widespread expression of Fos throughout neocortex. In somatosensory cortex, the induction of the c-fos gene by linopirdine was nearly completely blocked by atropine and scopolamine and strongly attenuated by the NMDA receptor blockers CPP and MK-801. The results suggest that the age-related decline in acetylcholine release in rodents can be partially compensated for by administration of linopirdine.     

IL-13对大鼠急性肾缺血再灌注时IL-1β表达的影响

目的:观察IL-13对急性肾缺血再灌注时IL-1β表达的影响。方法:Wistar雄性大鼠57只,随机分为8组:正常组(normal);假手术组(sham);缺血组:(I)缺血再灌注组(I/R);治疗对照组-1(C-1);治疗对照组-2(C-2);治疗组-1(T-1)和治疗组-2(T-2)。阻断大鼠双侧肾脏血流45min再灌注24h建立急性肾缺血再灌注模型;治疗组分别于阻断血流前、后分别从双侧肾动脉开口注射入1.5μg/50gbw鼠重组白细胞介素13(rmIL-13);检测各组大鼠IL-1β血清水平和肾脏表达,以及肾功能和肾脏病理。结果:(1)治疗组肾脏IL-1β基因(TtoC:P＜0.01)和蛋白表达(T-1toC-1:P＜0.01;T-2toC-2:P＜0.05)明显减少,血清IL-1β水平明显下降;(2)肾功能障碍和肾组织病理变化明显减轻,肾小管损害评分减少(C-1toT-1:45.20±8.64to21.05±8.82,P＜0.01;C-2toT-2:42.25±11.15to23.25±7.31,P＜0.01);(3)血清IL-1β水平与BUN、Cr成正相关(r=0.708,P＜0.01;r=0.770,P＜0.01)。结论:IL-13能有效地抑制大鼠急性肾缺血再灌注损伤IL-1β的表达。     

hCG对JEG-3滋养细胞系表达细胞因子的影响

目的：揭示人类绒毛膜促性腺激素（hCG)是否通过改变细胞因子的生成而影响滋养细胞的侵袭性。方法：以永生化的滋养细胞系JEG－3为研究对象，采用逆转录多聚酶链反应（RT－PCR）方法观察了hCG对JEG－3细胞与细胞侵袭力调节有关的多种因素因子表达的影响，结果：JEG－3细胞表达HGF，IGF－II，VEGF和TGF－β3，且VEGF的表达以VEGF121和VEGF165为主，而不表达IGF，TGF－1β，TGF－β2，IL－β1，25U／mL hCG处理50h可显著降低JEG－3细胞中HGF的表达，同时强烈诱导VEGF121和VEGF165的表达，而其它基因的表达未发生明显变化，结论：HGF对滋养细胞的侵入起促进作用，而VEGF则具有抑制效应，说明，高浓度的hCG可能通过两种细胞因子的自分泌机制对滋养细胞的侵入起抑制作用。     

散发性急性戊型肝炎血清抗体动态变化和肝脏超微…

为探讨散发性戊型肝为血清抗体动态变化，应用酶联免疫法（ＥＩＡ）检测了７例急性戊型肝炎抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ和ＩｇＭ抗体、并对１例２进行了肝超微结构病理检测，结果表明，发病１０天至４５天内抗ＨＥＶ－ＩｇＧ和ＩｇＭ滴度最高，发病第４０天仍有肝细胞肿胀，胞浆空化和线粒体固缩等病理变化。患者轿清抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月内全中消失，抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月     

Inflammatory cytokine response to Vibrio vulnificus elicited by peripheral blood mononuclear cells from chronic alcohol users is associated with biomarkers of cellular oxidative stress

Vibrio vulnificus is the leading cause of death in the United States associated with the consumption of raw seafood, particularly oysters. In epidemiological studies, primary septicemia and inflammation-mediated septic shock caused by V. vulnificus is strongly associated with liver disease, often in the context of chronic alcohol abuse. The present study was undertaken to determine whether clinical biomarkers of liver function or cellular oxidative stress are associated with peripheral blood mononuclear cell inflammatory cytokine responses to V. vulnificus. Levels of interleukin-1 beta (IL-1 beta), IL-6, IL-8, and tumor necrosis factor alpha elicited in response to V. vulnificus and measured in cell supernatants were not associated with the liver biomarkers aspartate aminotransferase (AST) or alanine aminotransferase (ALT) or the AST/ALT ratio. In contrast, reduced glutathione (GSH) levels were associated with the release of all four cytokines (IL-1 beta [R(2) = 0.382; P = 0.006], IL-6 [R(2) = 0.393; P = 0.005], IL-8 [R(2) = 0.487; P = 0.001], and TNF-alpha [R(2) = 0.292; P = 0.021]). Those individuals with below-normal GSH levels produced significantly less proinflammatory cytokines in response to V. vulnificus. We hypothesize that persons with markers for cellular oxidative stress have increased susceptibility to V. vulnificus septicemia.     

The effects of co-culture with human fibroblasts on human embryo development in vitro and implantation

In a human in-vitro fertilization (IVF) programme, the effect of co- culture of embryos with human fibroblasts was evaluated with respect to pregnancy rate and embryo development. Patients were included in the study after giving informed written consent. The IVF treatments were randomly assigned by stratification of both age (<36 versus > or =36 years) and previous IVF attempts (yes versus no). After fertilization was established, the zygotes were transferred to a 4-well dish with or without fibroblasts and cultured for 2 days. On the third day after ovum pick-up (OPU), cell number and quality [5 (good) to 1 (poor)] of the embryos were scored and a maximum of three embryos was transferred. Supernumerary embryos of good quality were cryopreserved. The design of this study was a group sequential trial with the objective of detecting differences between pregnancy rates following IVF with conventional incubation or incubation in co-culture with fibroblasts. This design included one evaluation at half-way data collection. In the study, 148 patients had an OPU, of whom 77 were allocated to the co-culture group. There was no statistically significant difference in pregnancy rate, cell number and embryo quality between the two groups. The ongoing pregnancy rate per embryo transfer was 27% in co-culture and 30% in the conventional culture group. The implantation rates per transferred embryo were 17 and 18% respectively. Using a multivariate logistic regression model for the probability of ongoing pregnancies, the odds ratio of co-culture, adjusted for age and previous IVF attempts, was not statistically significant. In conclusion, co-culture with human fibroblasts does not contribute to an improvement of embryo quality nor to a higher pregnancy rate after IVF in an unselected group of patients.      

肝素酶和碱性成纤维细胞生长因子与非小细胞肺癌转移和预后的关系

目的 探讨肝素酶(heparanase)和碱性成纤维细胞生长因子(bFGF)在人非小细胞肺癌(NSCLC)组织中表达的临床意义及其与肺癌转移和预后的关系。方法 采用免疫组织化学、原位杂交和Western blot方法,检测115例人NSCLC石蜡切片和45例新鲜肺癌及对应癌旁正常组织中肝素酶和bFGF的表达情况。采用χ^2检验、t检验、生存曲线和Cox比例风险回归等方法分析肝素酶和bFGF分别表达及共表达的意义。结果 免疫组织化学染色证实肝素酶(91／115)和bFGF(89／115)主要表达在癌细胞质和(或)细胞膜中,在正常肺泡上皮和支气管上皮中则呈阴性表达。Western blot也证实肝素酶在肺癌中的表达明显增高(P=0．041)。统计分析结果显示：肝素酶和bFGF的表达具有明显的一致性(P：0．0001),二者单独表达和共表达均与肺癌的分期、血管侵袭、淋巴结转移、微血管密度和预后有关,其中,二者共表达时与分期和微血管密度的相关性更显著;另外,bFGF还与肺癌的分化程度有关。多因素分析结果显示,肺癌的分化程度、血管浸润、淋巴结转移和bFGF的表达可以作为判断肺癌预后的危险因素,但肝素酶不是影响预后的独立因素。结论 肝素酶和bFGF均与肺癌的转移、血管生成和预后密切相关。     

Endothelin receptor expression in human decidua

The endothelins are signalling peptides that act via two receptors, ET(A) and ET(B). In the human endometrium, endothelin receptors have been demonstrated in glands and stroma and have been shown to vary during the course of the menstrual cycle. The present study was undertaken to determine whether or not expression of endothelin receptors changes during pregnancy or after administration of exogenous progestagens. The expression of the receptors was correlated with the appearance of basement membrane components during decidualization of the endometrial stroma. Decidual specimens (n = 15) were obtained during the first trimester of pregnancy and 10 at term. Sixteen pairs of endometrial biopsies were obtained from women with menorrhagia before and after exposure to exogenous progestagens. A total of 15 hysterectomy specimens were used as controls for the expression of stromal basement membrane proteins in the absence of decidualization. Autoradiography was carried out with selective ligands for ET(A) ([125I]-PD 151242) and ET(B) ([125I]-BQ3020). The distribution of ligand binding was then compared with the distribution of laminin alpha2 light chain and collagen IV. ET(A), ET(B), laminin alpha2 light chain, and collagen IV were expressed in stromal decidual cells in the first trimester of pregnancy. ET(B) was also found on endometrial glandular epithelium. Quantitative macro-autoradiography and multiple regression analysis demonstrated a highly significant positive correlation (P < 0.001) between expression of ET(B) and laminin alpha2 light chain. In the third trimester qualitative examination suggested a reduction of ET(A) in the stroma. Progestagen-induced decidua exhibited a similar pattern to that found in first trimester decidua. This study has demonstrated up-regulation of ET(B) during the progesterone- dependent process of decidualization and suggests a paracrine or autocrine role for endothelins in the decidua.