苦参碱对C6胶质瘤细胞的增殖和原癌基因表达的影响

目的:本研究拟探讨苦参碱对胶质瘤细胞株(C6细胞株)的抑制作用及其作用机理.方法:应用MTT比色法检测不同浓度苦参碱对C6细胞的增殖抑制,应用流式细胞仪观察苦参碱对C6细胞周期的影响,应用RT-PCR观察原癌基因C-myc表达的变化.结果:苦参碱对C6细胞增殖抑制作用呈剂量依赖性,当浓度为0.20g/L时,对C6细胞增殖的抑制率达到(53.8±5.6)%.流式细胞仪分析显示,用0.20g/L苦参碱培养3 d,细胞周期中G0/G1期所占比例增加,S期占细胞周期的比例降低.RT-PCR结果显示,随着苦参碱作用剂量的增加,C-myc基因的表达被明显抑制.结论:苦参碱对大鼠胶质瘤细胞系C6的增殖具有明显的抑制作用,并对原癌基因C-myc的表达具有抑制作用.     

急性自发性脑出血患者血清HA水平的表达情况及临床意义

目的 探讨急性自发性脑出血（ICH）患者血清HA水平的表达情况及临床意义。 方法 选取2016年4月～2017年8月我院收治的120例ICH患者为研究对象,并选取同期体检的102例健康者为对照组。采用改良Rankin 量表对ICH患者出院后3个月情况进行评分,根据评分分为预后良好者（≤2分）和预后不良者(＞2分),观察并分析患者透明质酸（HA）水平表达及临床意义。结果 预后良好者的年龄、血肿体积均明显低于预后不良者,差异有统计学意义（均P＜0.05）。入院时,ICH组患者HA水平明显高于对照组,差异有统计学意义（P＜0.05）。入院时,预后良好者美国国立卫生研究院卒中量表（NIHSS）评分及HA水平均明显低于预后不良者,且NIHSS评分与HA表达水平呈正相关（r=0.573,均P＜0.05）;治疗3个月后,mRS评分与HA表达水平亦呈正相关（r=0.757,P＜0.05）。单因素表示,预后良好者的年龄、血肿体积、HA水平、NIHSS评分及mRS评分与预后不良者比较,差异有统计学意义（P＜0.05）。Logistic多因素回归分析表示,年龄、血肿体积、HA水平、NIHSS评分及mRS评分与ICH患者预后不良相关。结论 ICH患者血清HA水平表达异常升高,病情严重,预后较差。因此,入院时血清HA水平的高表达可作为预后不良的预测因素。     

前列腺癌患者血清miRNA 194和miRNA 206表达水平及临床意义

【摘要】 目的 探讨前列腺癌（PCa）患者血清miRNA 194和miRNA 206表达水平及临床意义。方法 选取2008年5月～2014年12月我院收治的108例PCa患者为PCa组,以及同期就诊的60例良性前列腺增生（BPH）患者为BPH组、前列腺正常者60例为正常组,分析所有纳入研究者的临床及随访资料,记录年龄等一般临床资料和血清miRNA 194和miRNA 206等指标水平及PCa患者肿瘤及预后情况,并比较不同PCa患者间上述资料的差异性。结果 BPH组和PCa组患者血清miRNA 206水平较正常组低（P＜005）,而血清PSA水平和miRNA 194较正常组高(P＜005);且PCa患者其血清miRN A 206水平较BHP患者更低,而血清PSA和miRNA 194水平则更高（P＜005）;Gleason 评分大于7分、临床分期T3~T4期以及出现淋巴结转移和术后复发的PCa患者其血清miRNA 206水平均显著降低,而血清miRNA 194显著升高（P＜005）;死亡患者血清miRNA 206水平均显著低于生存者,而血清miRNA 194水平显著高于生存者（P＜005）;经 Log rank检验,Gleason 评分大于7分、临床分期T3~T4期、出现淋巴结转移、术后复发以及血清miR NA 194水平升高和miRNA 206水平降低的PCa患者其生存时间均较短（P＜005）;经多因素Cox 比例风险回归模型分析,淋巴结转移、术后复发以及血清miRNA 194升高和miRNA 206水平降低均为影响PCa患者预后的独立影响因素（P＜005）。结论 血清miRNA 194在PCa患者中显著上升,而血清miRNA 206水平显著下降,且血清miRNA 194的上升和miRNA 206的下降均为影响PCa患者预后的独立影响因素,因此临床上可将其作为评估PCa患者病情及预后的有效指标。     

Interplay between extracellular matrix components of Pseudomonas putida biofilms

The extracellular matrix of bacterial biofilms has at least two key functions: to serve as a structural scaffold for the multicellular community, and to play a protective role against external stress. In this work, we report a compensatory effect whereby Pseudomonas putida reacts to the lack of either of the two main surface proteins involved in biofilm formation, LapA and LapF, by increasing expression and production of a species-specific EPS. Elevated levels of the second messenger molecule cyclic di-GMP alter the balance of extracellular matrix components, and the phenotypes of lapA and lapF mutants under these conditions are indicative of direct interactions taking place between large secreted proteins and exopolysaccharides. Our data suggest the existence of a mechanism by which bacteria would sense alterations in the composition of the extracellular matrix, leading to changes in expression of the different elements.     

False-Positive β-Galactosidase Staining in Osteoclasts by Endogenous Enzyme: Studies in Neonatal and Month-Old Wild-Type Mice

Escherichia coli β-galactosidase (β-gal), encoded by the lacZ gene, has become an essential tool in studies of gene expression and function in higher eukaryotes. lac-Z is widely used as a marker gene to detect expression of transgenes or Cre recombinase driven by tissue-specific promoters. The timing and location of promoter activity is easily visualized in whole embryos or specific tissues using the cleavable, chromogenic substrate, 5-bromo-4-chloro-3-indolyl-D-galactopyranoside (X-gal). The tissue specificity of promoters in transgenic constructs is routinely tested by using a promoter of choice to drive lacZ. Alternatively, the targeted expression of Cre recombinase to perform in vivo recombination of loxP sites can be visualized by β-gal staining in mice carrying a Cre-activated lacZ transgene, such as the ROSA26 strain. In the course of our investigations, we examined β-gal activity in bone tissue from genetically normal mice using standard detection methodology and found very high endogenous activity in bone-resorbing osteoclasts. This was true in frozen, paraffin, and glycol methacrylate sections. X-gal staining colocalized with the osteoclast marker, tartrate-resistant acid phosphatase (TRAP). β-gal activity was present in osteoclasts in long bones, in the mandible, and in both neonatal and more mature animals. We present this brief article as a caution to those testing genetic models of skeletal gene expression using β-gal as a marker gene.     

Identification of Genes Differentially Expressed in Cultured Human Osteoblasts Versus Human Fibroblasts by DNA Microarray Analysis

Little is known about patterns of gene expression from cells populating the connective tissues. This study investigated the possible variance of gene expression profile between human osteoblasts (HO) and human fibroblasts (HF) in vitro, using DNA microarray technology. Clustering identification was used to compare expression patterns between HO and HF for biological significance. Our results showed that genes encoding the extracellular matrix or apoptosis-related proteins tended to be expressed in greater abundance in HO, while more proteolysis-related proteins were expressed in higher level in HF. Significant differences in expression were also noted with genes related to signaling pathways. To confirm the array results, three genes (periostin, MFG-E8, MMP-10) were selected and analyzed independently by RT-PCR and northern blot. The results were found consistent with the array data in HO and HF. The present findings suggest that HO and HF differ not only phenotypically but in the expression level of tissue specific genes to assure the turnover and homeostasis of their respective tissues.     

Gene Expression Omnibus资源库在肿瘤学研究中的应用

Gene Expression Ominibus （GEO）是供学者们分享基因表达谱芯片数据、高通量测序数据等的国际化的公共资源平台,其中涵盖了应用炎癌模型获得的大量相关实验数据,比如关于某些炎性因子的表达情况.这些数据为炎癌转化中的调控网络和分子机制的研究提供了帮助,也为肿瘤的防治提供了新思路.研究者针对新的预测基因进行研究时,起源于各种疾病模型或实验处理的GEO数据库为寻找其有关功能提供了有利条件和有力证据.     

Characterization and expression of the CXCR1 and CXCR4 in miiuy croaker and evolutionary analysis shows the strong positive selection pressures imposed in mammal CXCR1

The innate immune system can recognize non-self, danger signals, and pathogen associated molecular patterns and provides a first line of antimicrobial host defense. Therefore, it plays an instructive role and is pretty important in vertebrates. In innate immune responses, CXCRs act as the main receptors of CXC chemokines and play a vital role in host defense and inflammation. In present study, we cloned two cDNA molecules of CXCR1 and CXCR4 in Miichthys miiuy (miiuy croaker). In these two genes, we found the most highly conserved DRY motif in the second intracellular loop adjacent to the third transmembrane domain. The expressions of CXCR1 and CXCR4 showed that they were ubiquitously expressed in ten normal tissues. After infection with Vibrio anguillarum and Vibrio harveyi, the expressions of CXCRs in the immune tissues were significantly regulated in most of tissues except that of CXCR1 in the kidney after V. harveyi injection. Evolutionary analysis showed that only the ancestral lineages of CXCR4 in amphibians underwent positive selection, indicating that the ancestors of amphibians boarded the land and had to further evolve to adapt to terrestrial environments. Multiple ML methods were implemented to detect the robust positively selected candidates for sites. In total, we detected 12 and 3 positively selected sites in the subsets of current mammal and fish CXCR1 genes, and only one site under positive selection was found in mammalian CXCR4 subsets. These positively selected sites were mainly located in the extracellular domains of CXCRs. The sliding window analysis and evolution test tended to favor positive selection acting on the N-terminal domain of CXCR1, which was the critical region for ligand/receptor signaling for neutrophils and receptor–ligand interaction, indicating that the N-terminal of CXCR1 in mammals underwent more positive selection than that of fish.     

细胞黏附分子CD99在人髓母细胞瘤中的表达及意义

目的 探讨细胞黏附分子CD99在人髓母细胞瘤组织中的表达特点及其临床意义。方法 采用免疫组化法检测107例髓母细胞瘤及15例瘤旁正常脑组织标本中CD99的表达。所有病人均随访至死亡或截止至2013年12月，随访时间为12~80个月，利用Kaplan-Meier生存曲线进行生存分析。结果 髓母细胞瘤CD99阳性表达率（57.94%，62/107）明显高于瘤周正常脑组织（0%，0/15；P<0.05）。CD99的表达水平与病人性别、年龄、肿瘤的病理组织学分型和分子亚型均无明显相关性（P>0.05）。CD99的高表达与髓母细胞瘤病人的预后呈显著负相关（P<0.05）。结论 CD99高表达与髓母细胞瘤预后显著相关，可作为预后判断的指标及治疗髓母细胞瘤潜在的新靶点。