脓毒症患者外周血单核细胞miR-147b的表达及其与病情严重程度和预后的关系

目的 探讨脓毒症患者外周血单核细胞miR-147b的表达水平及其与病情严重程度及预后的相关性。方法 选择2019年1月至2021年6月于安徽医科大学第一附属医院就诊的46例脓毒症患者(脓毒症组)和50例普通感染患者(普通感染组)作为研究对象。查阅患者病历,记录年龄、性别、感染部位等一般资料。采用酶联免疫吸附试验试剂盒检测研究对象血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和C反应蛋白(C-reactive protein, CRP)的浓度。计算序贯性器官衰竭评分(sequential organ failure score, SOFA)和急性生理学与慢性健康状况评分Ⅱ(acute physiology and chronic health scoreⅡ,APACHEⅡ)。观察或随访脓毒症患者的28 d预后情况,分为生存组和死亡组。应用逆转录聚合酶链反应测定外周血单核细胞中miR-147b的表达水平。结果 脓毒症组和普通感染组的年龄、性别、感染部位分布比较差异无统计学意义(P>0.05)。脓毒症组患者的TNF-α、CRP水平、SOFA评分和A...     

miR-21靶向E2F1对三阴性乳腺癌细胞恶性生物学活性及裸鼠肿瘤抑制率的影响

目的 探究miR-21靶向E2F1对三阴性乳腺癌细胞恶性生物学活性及裸鼠肿瘤抑制率的影响。方法将MDA-MB-231细胞分为5组,即MDA-MB-231组、miR-21 inhibitor组、miR-NC inhibitor组、siRNA-E2F1组和siRNA-NC组。检测细胞中miR-21表达（RT-PCR法）;分别检测细胞增殖（MTT法）、侵袭（Transwell法）、迁移（划痕实验）和凋亡能力（流式细胞仪）;检测细胞中E2F1蛋白表达;检测miR-21与E2F1的靶向关系（双荧光素酶实验报告）。结果MDA-MB-231细胞中miR-21表达明显高于MCF10A细胞（P<0.05）;miR-21 inhibitor组细胞细胞中miR-21表达明显低于MDA-MB-231组（P<0.05）。与MDA-MB-231组相比,miR-21 inhibitor组细胞吸光度值、侵袭能力、迁移能力和细胞中E2F1蛋白表达均明显降低,细胞凋亡能力明显升高（P<0.05）;MDA-MB-231组细胞吸光度值、侵袭、迁移、凋亡能力和细胞中E2F1蛋白表达与miR-NC inhibitor组相比差异无统计学意义（P>0.05）。预测软件显示E2F1的3′UTR端与miR-21有碱基互补结合点位。通过向MDA-MB-231细胞中转染野生型E2F1（E2F1-WT）时,miR-21组荧光素酶活性明显低于miR-NC组（P<0.05）;miR-21组和miR-NC组突变体荧光素酶活性相比差异无统计学意义（P>0.05）。与siRNA-NC组相比,siRNA-E2F1组细胞增殖、侵袭、迁移和细胞中E2F1蛋白表达均明显降低,细胞凋亡能力明显增加（P<0.05）。与miR-NC inhibitor组裸鼠移植肿瘤第8天时相比,miR-21 inhibitor组裸鼠肿瘤体积明显降低,肿瘤抑制率为45.3%（P<0.05）。结论低表达miR-21可抑制三阴性乳腺癌细胞增殖、侵袭,促进凋亡,且抑制裸鼠移植瘤体积,其作用机制可能与抑制E2F1表达有关。     

The effect of intravenously administered salbutamol on serum potassium in asthmatic and nonasthmatic atopic subjects

The role of adrenergic mechanism in the pathogenesis of allergic disease is controversial. Recent experimental and clinical reports have suggested that -adrenergic blockade impairs and stimulation enhances extrarenal potassium uptake in humans. This led us to study the effect of the intravenous administration of salbutamol, a specific -2-adrenergic agonist, on serum potassium in 9 healthy subjects and in 23 patients with allergic asthma and/or rhinitis. Serum potassium fell significantly and reached a peak decline at the end of venous infusion in all the normal subjects. Seventeen atopic subjects showed a lower or absent serum K⁺ decrement: there was no difference between asthmatic and rhinitic patients. There was no relation among the salbutamol-induced serum potassium decrement, serum glucose increment, blood pressure and heart-rate changes, and nonspecific bronchial reactivity. These findings suggest that -2-adrenergic hyporesponsiveness is present only in some allergic patients.     

An improved method for the isolation of the major protein of the gonococcal outer membrane in an antigenically reactive form

The major outer membrane protein (protein I) has been isolated from Neisseria gonorrhoeae strain P9 in an immunologically reactive form. Membranes were sequentially extracted with the detergents sodium cholate and Empigen BB. Protein I was enriched in the Empigen-soluble fraction and was separated from other proteins and lipopolysaccharide by gel filtration chromatography on Sephadex G-200. The purified protein retained its antigenic activity with antiserum raised against the unfractionated outer membrane complex.     

Characterization of EN4 monoclonal antibody: a reagent with CD31 specificity.

EN4 MoAb was originally described as a MoAb that reacts specifically with human endothelial cells, and the reagent was not assigned to any of the presently known CD. Here, we provide evidence indicating that EN4 reacts with the CD31 antigen. Thus, EN4 stains strongly murine fibroblasts transfected with the human CD31 gene. Furthermore, SDS-PAGE analysis of immunoprecipitates of cell lysates from surface-iodinated Jurkart T cells demonstrated that EN4 and reference CD31 MoAb recognized the same antigen, of 130 kD mol. wt. Finally, both EN4 and CD31 gave the same pattern of reactivity when tested on tonsillar or peripheral blood lymphoid cells by FACS analysis or by immunohistochemistry on sections of a variety of human tissues. EN4, however, proved consistently more efficient than the reference anti-CD31 MoAb as judged by both the intensity of fluorescence or of tissue staining. This property has thus allowed a better characterization of the tissue and cellular distribution of CD31.     

Functional mapping of the activity of the R region in the human T-cell leukemia virus type I long terminal repeat to increase gene expression

We previously demonstrated the activity of the R fragment in the long terminal repeat of human T-cell leukemia virus type I for elevation of the level of gene expression. In this study, the fragment was deleted with BAL31 nuclease to determine its functional domain. Series of the shortened R fragments were linked to the simian virus 40 promoter unit, which regulated expression of a reporter gene. Examination with the R fragments deleted from the 5 and 3 ends showed that borders of the functional domain were mapped within nucleotide positions 458 to 473 for the 5 end and nucleotide positions 559 to 594 for the 3 end, respectively. Thus we conclude that a 136-base-pair fragment corresponding to the second half of the R region was sufficient to allow elevation of the level of gene expression.     

人IL-31基因克隆、表达及在皮肤炎症中的作用

目的 克隆人IL-31基因,构建原核表达载体,并诱导其在大肠杆菌中表达,研究hIL-31蛋白与皮肤炎症的关系.方法 采用RT-PCR克隆人IL-31基因,将其克隆到原核表达载体pET28a( ),并通过IPTG诱导目的基因在大肠杆菌中表达,用该目的蛋白刺激人表皮角质细胞HaCaT,RT-PCR检测趋化因子MIP-3β、TARC及TCA-3(I-309)mRNA的表达;小鼠皮内注射该目的蛋白,观察局部炎症表现,皮肤标本HE染色观察皮肤炎性特征,采血进行白细胞计数及分类,ELISA法检测血清IL-1βIL-6和TNF-α水平,流式细胞仪分析胸腺及脾脏T细胞亚群.结果 成功获得495bp的人IL-31基因,原核表达质粒构建正确,IPTG诱导目的基因在大肠杆菌中大量表达;该目的蛋白可刺激HaCaT表达趋化因子MIP-3β、TARC、I-309;小鼠皮肤注射部位有脱毛,HE染色可见炎性细胞浸润,外周血白细胞总数增加,中性粒细胞比例增高,血清IL-1β、IL-6和TNF-α水平增高,脾脏CD4 T细胞百分比增高,CD8 T细胞百分比减少.结论 人IL-31基因克隆及原核表达已获成功,hIL-31蛋白可刺激HaCaT细胞表达MIP-3β、TARC、I-309,诱导小鼠皮肤炎症反应.     

The significance of angiogenesis in malignant pheochromocytomas

The purpose of this study was to investigate tumor angiogenesis in a series of benign and malignant pheochromocytomas and to determine whether there is a correlation between angiogenesis and the presence of distant metastases. In this study, the CD31 monoclonal antibody was selected to measure intratumoral microvessel density. Nineteen patients with malignant pheochromocytomas and nineteen patients with benign pheochromocytomas who underwent operation were studied. In order to quantify intratumoral microvessel density, the total number of pixels of CD31-positive reactivity was assessed and expressed as a percentage of the total tissue area in the analyzed field. Analysis of variance revealed a statistically significant correlation between malignancy and intratumoral microvessel density (p=0.0009). Although there was a considerable variability in the intratumoral microvessel density from tumor to tumor within both the benign and the malignant group, a percentage of more than 28.5% anti-CD31 stained area was found only in malignant tumors. In conclusion, this study shows that the mean intratumoral microvessel density in malignant pheochromocytomas is increased approximately two-fold as compared with benign tumors. However, the clinical significance of this prognostic marker is rather weak, because only 4 of the 19 malignant pheochromocytomas had microvesel density higher than this threshold of 28.5%.     

<Emphasis Type="Italic">GPC5</Emphasis> is a possible target for the 13q31-q32 amplification detected in lymphoma cell lines

Comparative genomic hybridization (CGH) analyses have detected gains of copy number on 13q, especially at 13q31-q32, in cell lines and primary cases of various types of lymphoma. Since amplification of chromosomal DNA is one of the mechanisms that can activate tumor-associated genes, and because 13q amplification had been reported in various other types of tumors as well, we attempted to define by fluorescence in situ hybridization (FISH) a common region at 13q31-q32 in which to explore genes that might be targets for the amplification events. Although the commonly amplified region we defined was relatively large (approximately 4 Mb), only one true gene, GPC5, was found there. GPC5 was over-expressed in lymphoma cell lines that had shown amplification, in comparison with those that had not. Our findings suggest that GPC5 is a likely target for amplification, and that over-expression of this gene may contribute to development and/or progression of lymphomas and other tumors.     

Overexpression of TONSL might be an independent unfavorable prognostic indicator in hepatocellular carcinoma

Background

TONSL has been suggested to function as an oncogene in lung, esophageal and cervical cancer. This study was aimed to identify the expression of TONSL and its role in hepatocellular carcinoma (HCC).