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The expression levels of heparanase mRNA, uPA mRNA and PAI-1 mRNA were analyzed semiquantitatively, using the reverse transcriptase-polymerase chain reaction method, and were compared with clinicopathological findings.

二者相比具有显著性差异（P ：肝素酶mRNA、b-FGF和uPA在膀胱癌组织中的表达上调，三者在膀胱癌的发生、发展过程中具有重要作用。

Five rats in both groups were executed at 3, 7, 14, and 28 days after model establishment. RNA was extracted from the same site of left anterior wall, and the polymerase chain reaction was used to semiquantitatively analyze the Ang1 and Tie2 receptor mRNA expression with GAPDH gene as internal control; meanwhile, the immunohistochemistry was used to detect vascular density in and around infarction area.

于术后3，7，14，28 d 四个时间点，每组取5只处死，取心脏左室前壁同一部位，提取总RNA，用RT-PCR的方法，以GAPDH基因为内参，进行半定量分析检测正常心脏及梗死后血管生成素1及Tie2 mRNA的表达；同时用免疫组化的方法检测各时间点梗死区域以及梗死周边区域血管数量。

The results showed that RSAP used two primers of 18 nucleotides in which starting at the 3' end of each primer were restriction site sequences (4-6 bases),followed 12-14 bases of arbitary sequence, the differences between two primers were restriction sites and filler sequence; PCR am-plification was run for the first 5 cycles with an annealing temperature of 35℃, followed by 35 cycles with an annealing temperature of 48℃; the PCR reaction of 25 μL included 20ng DNA templates, 2.5mmol/L of Mg(superscript 2+), 0.2 mmol/L of dNTP, 1.5U of Taq DNA polymerase, 600nmol/L of each primers. RSAP is a new maker technique with simplicity, moderate throughput ratio and reliability. RSAP is of good reprodicibity and broad application.

结果显示，RSAP技术的引物为2条长度均为18bp的引物，引物的3'端为4~6个碱基的限制性酶切位点序列，接着是12~14个碱基的随机序列，2条引物的限制性位点和随机序列不同：PCR扩增的前5个循环采用35℃的退火温度，随后的35个循环采用48℃的退火温度；在25μL反应体系中，模板DNA用量为20ng，Mg(上标 2+)浓度为2.5mmol/L dNIPs浓度为0.2 mmol/L，Tap DNA聚合酶用碳为1.5U，2条引物浓度均为600mmol/L；RSAP技术重复性好，适用性广泛，是一种操作简便、产率中等、稳定可靠的DNA标记技术。

METHODS HBV DNA in sera of 205 patients with HBV infection was detected by polymerase chain reaction.

应用PCR技术检测不同HBV感染205例，患者血清HBV DNA，并与正常人20例作比较。

In order to establish a sensitive immuno-PCR assay for detecting serai anti-P53antibodies, investigate the relation of anti-P53 antibodies in patients with breast carcinoma to P53 protein expressions in tissue and clinical significances and afford a practical tool in diagnosis of breast carcinoma clinically. SaHIgG was coupled to PNCa DNA by chemical cross-linker PEI and SaHIgG-DNA probe was constructed.

本研究建立了血清抗P53蛋白抗体免疫聚合酶链反应(Immuno-polymerase chain reaction，免疫PCR)检测方法，探讨了乳腺癌患者血清中抗P53蛋白抗体与组织中P53蛋白表达之间的关系及临床意义，旨在为临床乳腺癌P53蛋白检测提供实用的替代工具。

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

Objective To explore a simple and quick polymerase chain reaction method for the genotyping of Fmr1 knockout mice.

目的为Fmr1基因敲除小鼠探索快速、简单的基因型PCR检测方法。

Methods The template DNA was extract from the human osteosarcoma cells line U－2OS by the single- step isolation method with isothiocyanic acid guanidine, the cDNA coding for the mature fragment of BMP－4 was amplified by the reverse transcription- polymerase chain reaction.

方法采用异硫氰酸胍一步法从人骨肉瘤细胞株U-2OS中提取总RNA，通过逆转录-聚合酶链反应扩增为BMP-4成熟区cDNA基因片段。

Methods The mRNA and protein expression levels of NCAM were detected by reversed transcriptive polymerase chain reaction and Western blot after the rat hippocampal neurons were incubated with 20, 40 and 80μg/ml sodium fluoride for 24 hours in vitro.

原代培养海马细胞暴露于20、40、80μg/ml氟化钠24 h后，用RT-PCR和Western blot方法分别检测细胞内NCAM mRNA和NCAM各蛋白亚型的表达。

Weeks later, all the rats were killed, the levels of sodium pump α1-,α2-, and α3-subunit mRNA and protein in the cortex of kidney were detected with reverse transcription polymerase chain reaction method and immunohistochemical assay, respectively.

分别应用RT-PCR及免疫组化技术，探讨1k1c高血压大鼠肾脏皮质钠泵α1、α2及α3亚单位mRNA及蛋白水平基因表达的改变。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。