Differential induction responses of delta-aminolevulinate synthase mRNAs during erythroid differentiation: use of nonradioactive in situ hybridization.

Expression of mRNAs encoding the erythroid-specific delta-aminolevulinate synthase (ALAS-E) and the nonspecific delta-aminolevulinate synthase (ALAS-N) were examined in murine Friend virus-transformed erythroleukemia (MEL) cells using nonradioactive in situ hybridization. Following dimethyl sulfoxide (DMSO) treatment, ALAS-E mRNA increased markedly, while ALAS-N mRNA did not increase in wild-type MEL cells. In contrast, in a DMSO-resistant clone of MEL cells, ALAS-E was not detectable before and after DMSO treatment. These findings suggest that ALAS-E and ALAS-N mRNAs are under separate controls and that the expression of ALAS-E mRNA is a critical event in erythroid differentiation.     

血管紧张素Ⅱ对大鼠心室肌三种离子通道基因表达的效应

目的利用微渗透泵灌注血管紧张素Ⅱ(AngⅡ)大鼠模型,观察AngⅡ对大鼠心室肌三种离子通道基因表达的效应。方法Sprague-Dawley大鼠48只植入微渗透泵,随机分为对照组灌注生理盐水(1μl/h),实验组灌注AngⅡ(200或400ng·kg-1·min-1),持续时间为2w或4w。提取心脏左室RNA,逆转录-聚合酶链反应半定量分析SCN5A、Kv4.3及Cav1.2离子通道基因mRNA的表达量,磷酸甘油醛脱氢酶(GAPDH)为内参。结果SCN5A及Cav1.2基因的mRNA水平则较对照组升高(P<0.05),而Kv4.3基因的mRNA水平较对照组降低(P<0.01),且具有一定剂量及时间依赖性。结论AngⅡ使SCN5A及Cav1.2表达上调,Kv4.3表达下调,且其效应具有剂量及时间依赖性。     

Wnt-1诱导分泌蛋白1在直肠癌中的表达及其临床意义

目的定量检测Wnt-1诱导分泌蛋白1（Wnt-1 induced secreted protein-1，WISP-1）基因在人直肠癌及远端正常直肠组织中的表达，为进一步研究该基因功能提供线索。方法提取人直肠癌及癌旁正常直肠组织总RNA，采用实时RT—PCR分析WISP-1的表达，再对PCR结果进行分析。结果WISP-1 mRNA在直肠癌组织中的表达水平显著高于癌旁正常直肠组织，差异有统计学意义（P〈0．05）。WISP-1的表达与肿瘤的分化程度、Dukes分期及淋巴结转移等因素有相关性。结论WISP-1基因的异常表达可能与直肠癌的发生、发展有关，有望成为直肠癌诊断预后的标志物和治疗的新靶点。     

用SYBR Green I实时逆转录-聚合酶链反应定量检测人、小鼠精子中CatSper1 mRNA

目的:建立并优化SYBR GreenI实时RT-PCR体系,定量检测人、小鼠成熟精子中的CatSper1 mRNA。方法:用TRIzol分别提取人、小鼠成熟精子中的总RNA,逆转录后用SYBR GreenI实时PCR定量检测CatSper1 mRNA。SYBR GreenI实时PCR采用普通PCR试剂,加入SYBR GreenI染料,优化退火温度、Mg2+浓度及上、下游引物比例,并在PCR循环时采用四步法以消除引物二聚体的影响。优化完成后用不同浓度的精子cDNA为模板做标准曲线,以检测SYBR GreenI实时PCR的扩增效率。结果:定量检测CatSper1 mRNA的SYBR GreenI实时PCR体系适宜退火温度、Mg2+浓度及上、下游引物比例分别为63℃、3.0mmol/L和1∶1,四步法中采集荧光的温度为88℃。优化后用人和小鼠精子cDNA为模板做标准曲线分别为Y=-3.402log(X)+25.99和Y=-3.409log(X)+24.09,扩增效率分别为96.8%和96.5%,可定量检测人、小鼠成熟精子中的CatSper1 mRNA。结论:用普通的逆转录及PCR系统和试剂,建立了一种方便、廉价、可靠的SYBR GreenI实时荧光定量RT-PCR系统,可用于人、小鼠精子中CatSper1 mRNA定量检测。     

c-met mRNA在乳腺癌术后腋窝引流液中表达的意义

目的探讨c-met mRNA在乳腺癌患者术后腋窝引流液中表达的意义。方法用RT—PCR法检测乳腺癌患者术后腋窝引流液中c-met mRNA的表达，分析其表达与肿瘤大小、腋窝淋巴结转移、雌激素受体（estrogen receptor，ER）、孕激素受体（progesterone receptor，PR）及生长因子受体2（c—erbB-2）的关系，以及5-FU加蒸馏水冲洗手术野对c—met mRNA表达的影响。结果①c—met mRNA能在乳腺癌术后腋窝引流液中表达，其阳性率高于病理检查腋窝淋巴结转移的阳性率（P〈0．05）。②腋窝淋巴结转移、肿瘤大小与c—met mRNA的表达有关。③肿瘤ER、PR及c—erbB-2的表达与c—met mRNA的表达无关。④5-FU加蒸馏水冲洗手术创面能明显降低c-met mRNA的表达。结论c—met mRNA是乳腺癌微转移研究的理想特异性标志物，与常规病理检查腋窝淋巴结相比，RT—PCR检测腋窝引流液中c—met mRNA的表达能更早检测出肿瘤细胞在胸壁淋巴管中的转移，并具有更高的准确性。     

TNF-α在沙漠干热环境下猪腹部肠管火器伤后肝组织中的表达

目的探讨沙漠干热环境下猪腹部肠管火器伤后肝脏组织TNF-α的表达和肝功能的变化。方法沙漠干热环境组和常温环境组健康长白仔猪各42头随机等分为对照组和伤后1、2、4、8、12 h和24 h组,实验组建立腹部火器伤肠管穿透模型后,用免疫组化及图像分析法测定各组肝脏组织内TNF-α的表达,同时测定血清中AST水平。结果伤后各组肝组织TNF-α表达明显高于对照组,沙漠干热环境组于伤后2 h和8 h出现2个高峰;常温环境组于伤后2 h和12 h出现2个高峰。血清AST水平于伤后显著增高,并于伤后2 h出现第1个高峰,沙漠干热环境组和常温环境组分别于伤后8 h和12 h出现第2个高峰。结论沙漠干热环境下腹部肠管火器伤后肝脏TNF-α的表达增加并与AST的变化趋势一致,而且峰值较常温组提前出现,提示TNF-α在腹部肠管火器伤后肝损伤过程中可能起重要作用。     

黄芪对ESWL致兔肾损伤后缺氧诱导因子-1α表达的影响

目的探讨缺氧诱导因子-1α（HIF-1α）在ESWL致肾脏损伤中的作用及黄芪是否通过调节HIF-1α的表达而发挥对肾脏的保护作用。方法45只家兔随机分为对照组、ESWL损伤组和黄芪治疗组,每组15只,除对照组外,所有动物接受ESWL处理（18kV,1 500次）。黄芪治疗组家兔于冲击前3d至后2d,每天注射黄芪注射液2.0g/kg。2周后处死家兔,取肾脏HE染色观察肾组织形态学改变,免疫组织化学染色观察HIF-1α在肾脏中的表达。结果ESWL处理后肾小管上皮细胞肿胀、脱落,小管内可见大量管型,肾间质大量炎细胞浸润;黄芪治疗组以上病变显著减轻。ESWL损伤组HIF-1α显著表达于肾小管上皮细胞及间质,黄芪能够明显抑制HIF-1α的表达。结论HIF-1α参与了ESWL对肾脏的损伤,黄芪可能通过抑制HIF-1α的表达发挥对肾脏的保护作用。     

Differential Cellular Gene Expression in Ganglioglioma

Summary:  Purpose:  Gangliogliomas (GGs) are neuronal-glial tumors highly associated with epilepsy. We hypothesized that the expression of select gene families including neurotransmitter receptor subunits and growth factors would be distinct in neurons and astrocytes within GG compared with adjacent cortex and that these changes would yield insights into seizure onset and lesion formation.
Methods:  Candidate gene expression was defined in single immunohistochemically labeled neurons and astrocytes microdissected from GG specimens compared with neurons and astrocytes microdissected from morphologically intact cortex adjacent to the GG or normal control cortex.
Results:  Differential expression of 16 genes including glutamate transporter (EAAC1) and receptor (NMDA2C, mGluR5), growth factor (hepatocyte growth factor), and receptor (platelet derived growth factor receptor β, fibroblast growth factor receptor 3) mRNAs was detected in GG neurons compared with control neurons. In astrocytes, altered expression of p75NGF, mGluR3, TGFβ3 and Glt-1 mRNAs was detected. Nestin mRNA, a gene that exhibits enhanced expression in balloon cell cortical dysplasia, was increased in GG neurons. Because of the morphological similarities between GG and cortical dysplasia, we show that there is activation of the mTOR cascade in GG as evidenced by enhanced expression of phospho-p70S6kinase and phosphoribosomal S6 proteins.
Conclusion:  We find differential candidate gene expression in neurons and astrocytes in GG compared with adjacent cortex and show that there is activation of the mTOR pathway. These changes highlight pathways that may be pivotal for epileptogenesis and lesion growth.