荧光定量检测乙肝病毒含量在PHC患者中的临床应用

为进一步探讨HBV感染与原发性肝癌（PHC）的关系,本文采用实时荧光定量PCR法（Fluorescence quantitative PCR,FQ—PCR）对238例PHC患者进行HBV—DNA定量分析,旨在了解PHC患者HBV的定量情况,探讨HBV—DNA定量检测的应用价值,以及与乙肝标志物（HBV—M）的关系。     

实时荧光定量PCR法检测人骨肉瘤耐MTX细胞系中RFC、DHFR、GST-π的mRNA表达

[目的]研究RFC(还原叶酸载体)、GST-π(谷胱甘肽S转移酶π)、DHFR(二氢叶酸还原酶) mRNA在人骨肉瘤U2-OS细胞系及人骨肉瘤耐药细胞系U2-OS/R1-R3中的表达差异,并探讨其在人骨肉瘤MTX化疗耐药中的意义.[方法]采用冲击并逐步增加诱导药物浓度诱导的方法,诱导人骨肉瘤细胞系U2-OS,并建立耐MTX细胞系(U2-OS/R1-R3),利用荧光定量PCR技术在mRNA 水平检测RFC、GST-π、DHFR在U2-OS及(U2-OS/R1-R3)中的表达.[结果]本试验成功建立三株人骨肉瘤耐药细胞系U2-OS/R1-R3中,荧光定量PCR结果显示MTX耐药与DHFRmRNA表达增加有关,与RFCmRNA的表达降低有关,与 GST-πmRNA的表达增加有关.[结论]本试验在基因水平探讨人骨肉瘤细胞MTX化疗耐药机制, DHFR、GST-π基因表达的增加及RFC基因表达的降低参与了人骨肉瘤细胞的MTX耐药,为临床探索骨肉瘤患者的MTX耐药机制提供了重要的依据,为临床筛选MTX化疗不敏感患者提供重要的依据.     

原发性胆汁性肝硬化患者血清中miR-132和miR-212的表达及意义

目的探讨原发性胆汁性肝硬化(PBC)患者血清中微小RNA-132(miR-132)和miR-212水平的变化及其临床意义。方法收集PBC患者、其他肝病患者和健康对照组各37例血清标本;用实时荧光定量聚合酶链反应(RT-qPCR)检测miR-132和miR-212的水平,并分析其与疾病严重程度的相关性。结果与健康对照组及其他肝病组比较,PBC患者血清miR-132和miR-212的水平显著降低(P均0.05)。PBC组血清miR-132与ALP、AST呈负相关(r=-0.67,r=-0.61,P均0.05);而血清miR-212与ALP、AST无相关性(r=-0.34,r=-0.25,P均0.05)。结论 miR-132和miR-212可能参与了PBC的发病机制,有望作为PBC的诊断和治疗靶点。     

THP-1源性巨噬细胞对卵巢癌SKOV3细胞Toll样受体mRNA表达的影响

目的研究人单核细胞THP-1细胞系源性M2型巨噬细胞与卵巢癌SKOV3细胞相互作用后,肿瘤细胞Toll样受体(Toll-like receptors,TLRs)mRNA表达水平的改变。方法用佛波酯(PMA)诱导THP-1细胞分化为巨噬细胞,用免疫荧光技术检测THP-1细胞膜表面CD206分子的表达;用Transwell小室建立巨噬细胞与卵巢癌SKOV3细胞体外非接触式共培养模型;相互作用24 h后提取SKOV3细胞总RNA,用实时荧光定量PCR(real-time PCR)方法检测TLR1~TLR9 mRNA的表达水平并通过相对定量法(2-△△Ct)进行分析。结果 PMA诱导后,THP-1细胞巨噬细胞化,CD206分子高表达;卵巢癌细胞经共培养后TLR2、TLR3、TLR4、TLR5 mRNA表达水平显著增高,2-△△Ct分别为25.99±1.09、7.11±0.27、16.92±0.11、44.80±0.78。结论卵巢癌SKOV3细胞与M2型巨噬细胞相互作用后,肿瘤细胞TLR2~TLR5 mRNA表达显著增加,可能参与卵巢癌发生、发展过程。     

rhGDNF对大鼠脊髓损伤后Ntyr表达影响的研究

目的研究大鼠脊髓损伤（spinal cord injury,SCI）后应用人重组胶质细胞源性神经营养因子（recombinated human glial cell line derived neurotrophic factor,rhGDNF）对酪氨酸硝基化蛋白（nitrotyrosine,Ntyr）表达情况的影响。方法48只雄性SD大鼠随机分为正常组、假手术组、生理盐水对照组和rhGDNF实验组,改良Allen法致脊髓中等程度损伤,伤后1d、3d、7d、14d用免疫荧光染色法测定Ntyr的表达情况,同时评定下肢运动功能（Tarlov评分和改良Rivlin斜板）。结果伤后1d rhGDNF组Ntyr阳性神经元数量多于生理盐水对照组（P〈0．05）,伤后3d、7d、14d时较同时程生理盐水对照组明显减少（P〈0．05）。2组的运动功能评分比较：伤后1d无明显差异,伤后3d、7d、14d时rhGDNF治疗组较生理盐水对照组明显提高（P〈0．05）。结论SCI后应用rhGDNF可减少大鼠脊髓Ntyr的表达,促进其运动功能恢复。     

Axonal regeneration from the spinal cord to peripheral nerve induced by end-to-side neurorrhaphy Evidence from acetylcholinesterase staining and Fluorogold retrograde tracing

BACKGROUND： In recent years, surgeons have advocated root or trunk repair of avulsed nerve roots for overall recovery. However, donor nerves pose a major problem, because they do not contain adequate numbers of axons. Moreover, the procedures lead to nerve deficits in the donor nerve following transplantation. OBJECTIVE： To observe whether axonal regeneration occurs by end-to-side neurorrhaphy in the peripheral nerve and spinal cord. DESIGN, TIME AND SETTING： A neuroanatomical, randomized, controlled, animal study was performed at Functional Anatomy Lab in Nagoya University School of Medicine from May 2002 to July 2003. MATERIALS： Fluorogold was purchased from Fluorochrome, LLC, USA. BX50 light microscope and fluorescent microscope were purchased from Olympus, Japan. METHODS： A total of 21 rats were randomly divided into three groups, and the posterior avulsion injury model （C6-8） of the brachial plexus was performed. In the ventral root graft group, the avulsed C7 ventral roots were reanastomosed to the small anterior lateral aspect window of the spinal cord via nerve grafts. In the dorsal root graft group, the C7 dorsal roots were reanastomosed at the small pia mater window of the posterior lateral aspect of the spinal cord via nerve grafts. In the control group, the avulsed nerve roots were not repaired. MAIN OUTCOME MEASURES： The nerve grafts were collected from the ventral and dorsal root graft groups, and the C7 proximal nerve end was collected from the control group. Acetylcholinesterase staining was performed on the tissue. Fluorogold retrograde tracing technique was applied to determine the origin of the regenerating axons. RESULTS： Results showed that acetylcholine-positive axons existed in nerve grafts of the ventral and dorsal root graft groups. However, axons were not found in the avulsed nerve roots of the control group. Fluorogold retrograde tracing confirmed the presence of fluorogold-containing neurons in the ventral and dorsal horn of the ventral and dorsal root graft groups. Fluorogold-positive neurons were not observed in the control group. CONCLUSION： End-to-side neurorrhaphy induced axonal regeneration from the spinal cord to the peripheral nervous system.     

γ干扰素对实验性肝纤维化大鼠Smad3 mRNA的影响

目的初步探讨γ干扰素抗纤维化的分子机制。方法SD大鼠70只,模型对照组和γ干扰素治疗组注射40?l4油剂按0.3ml/100g皮下注射,2次/周,另设正常对照组大鼠10只。γ干扰素治疗组造模同时肌内注射γ干扰素0.2MU/kg,1次/日,模型对照组及正常对照组肌内注射生理盐水,连续干预12周。第12周末处死所有大鼠。取肝脏标本行常规苏木素-伊红和胶原染色,在光镜观察肝组织炎症活动度和肝纤维化情况并进行评分;实时荧光定量PCR方法检测大鼠肝脏Smad3mRNA的表达。结果肝组织炎症活动度和肝纤维化评分显示γ干扰素治疗组大鼠炎症程度、肝纤维化程度与模型对照组大鼠的肝组织炎症程度、肝纤维化程度比较显著改善(P<0.01,P<0.01)。荧光定量RT-PCR检测显示:与正常对照组Smad3mR-NA相比,模型对照组大鼠肝脏中Smad3mRNA表达显著增加(P<0.01);与模型对照组相比,γ干扰素组大鼠肝脏组织中Smad3mRNA表达明显减少(P<0.05)。结论γ干扰素抗实验性肝纤维化作用与下调Smad3mRNA来实现。干预转化生长因子β1信号转导过程是γ干扰素抗肝纤维化的机制之一。     

眼底彩照和荧光照影在初诊2型糖尿病患者的临床应用

目的探讨眼底彩照和荧光照影在初诊2型糖尿病患者的眼底检查中的临床意义。方法采用眼底照相和荧光照影对102例（204只眼）的初诊为2型糖尿病患者的眼底检查、诊断。结论眼底彩照和荧光照影可以尽早地发现和治疗糖尿病视网膜病变,又可以为内科医生反映出全身微血管的变化情况,具有重要的的临床意义。     

The Development ofA Fluorescence Polarization Immunoassay for Aflatoxin Detection

<正>A fluorescence polarization immunoassay(FPIA)was developed for the analysis ofaflatoxins(AFs)using an anti-aflatoxin B1(AFB1)monoclonal antibody and a novel fluorescein-labeled AFB1tracer.The FPIA showed an IC50 value of 23.33ng/mL with a limit of detection of 13.12 ng/mL for AFB1.The cross-reactivities of AFB1,AFB2,AFG1,AFG2,AFM1,and AFM2 with the antibody were100%,65.7%,143%,23.5%,111.4%,and 2%,respectively.The group-specificity of anti-AFB1mAb indicated that the FPIA could potentially be used in a screening method for the detection of total AFs,albeit not AFG2 and AFM2.The total time required for analyzing 96 samples in one microplate was less than 5 min.This study demonstrates the potential usefulness of the FPIA as a rapid and simple technique for monitoring AFs.