目的:比较重组组织型纤溶酶原激活剂(Recombinant human tissue-type plasiminogen,rt-PA)与尿激酶溶栓后对基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)表达和脑出血性转化的影响。方法:采用大鼠自体血栓塞大脑中动脉模型,缺...目的:比较重组组织型纤溶酶原激活剂(Recombinant human tissue-type plasiminogen,rt-PA)与尿激酶溶栓后对基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)表达和脑出血性转化的影响。方法:采用大鼠自体血栓塞大脑中动脉模型,缺血6h后静脉应用rt-PA及尿激酶,24h后处死,脑组织中MMP-9表达采用免疫组织化学法,脑出血量的测定采用分光光度计法。结果:脑缺血后MMP-9表达增加,rt-PA与尿激酶溶栓后MMP-9表达均进一步增加。脑缺血对照组、rt-PA溶栓组及UK溶栓组平均光密度值分别为0.158±0.010、0.225±0.0341、0.228±0.022,差异有统计学意义(P<0.01)。溶栓后两组脑出血发生率及脑出血量均较缺血对照组增加,但两组之间无明显差别。结论:rt-PA与尿激酶溶栓后均可上调MMP-9表达并与出血性转化密切相关。展开更多
Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform ...Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp （green fluorescent protein） and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone （AS） was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques （RT-PCR） and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis （SE） decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.展开更多
Summary: Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gen...Summary: Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis. pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using irnmunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected.展开更多
Objective To investigate the aberrant expressions of TNF-α and TGF-β1 in peripheral blood mononuclear cells （PBMCs） and placental tissues in patients with early spontaneous abortion Methods Using the technique of ...Objective To investigate the aberrant expressions of TNF-α and TGF-β1 in peripheral blood mononuclear cells （PBMCs） and placental tissues in patients with early spontaneous abortion Methods Using the technique of semi-quantitative reverse transcript-polymerase chain reaction （RT-PCR）, TNF-α mRNA and TGF-β1 mRNA in PBMCs were measured in spontaneous abortion group （30 cases）, normal pregnancy group （25 cases） and nonpregnant group （25 cases）. The expressive intension of TNF-α protein and TGF-β1 protein in placental tissues was also identified by immunohistochemistry. Results Both levels of TNF-α mRNA and TGF-β1 mRNA expressed in PBMCs were significantly different between the three groups respectively （P〈0.05）. Levels of TNF-α in syncytiotrophoblastic and cytotrophoblastic cells of the two aborted groups were substantially higher than those of the nonregnant group （P〈0.01）, but the levels of TGF-β1 in syncytiotrophoblastic cells of the two aborted groups were markedly lower than those of the non-pregnant group （P〈0.01）. Conclusion There is potential relation between TGF-β at the fetomaternal interface and spontaneous abortion. TGF-β1 may contribute to the maintenance of pregnancy, and low-level expression of TGF-β1 may be associated with pregnancy failure.展开更多
文摘目的:比较重组组织型纤溶酶原激活剂(Recombinant human tissue-type plasiminogen,rt-PA)与尿激酶溶栓后对基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)表达和脑出血性转化的影响。方法:采用大鼠自体血栓塞大脑中动脉模型,缺血6h后静脉应用rt-PA及尿激酶,24h后处死,脑组织中MMP-9表达采用免疫组织化学法,脑出血量的测定采用分光光度计法。结果:脑缺血后MMP-9表达增加,rt-PA与尿激酶溶栓后MMP-9表达均进一步增加。脑缺血对照组、rt-PA溶栓组及UK溶栓组平均光密度值分别为0.158±0.010、0.225±0.0341、0.228±0.022,差异有统计学意义(P<0.01)。溶栓后两组脑出血发生率及脑出血量均较缺血对照组增加,但两组之间无明显差别。结论:rt-PA与尿激酶溶栓后均可上调MMP-9表达并与出血性转化密切相关。
基金supported by the National Natural Science Foundation of China (30471212,30500347)the Earmarked Fund for Modern Agro-Industry Technology Research System,Ministry of Agriculture,China (NYCYTX-3-CY-04)
文摘Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp （green fluorescent protein） and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone （AS） was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques （RT-PCR） and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis （SE） decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.
文摘Summary: Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis. pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using irnmunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected.
基金This study is supported by the Fund of Fujian Provincial Health Department for the Transcenturial Scholarship Leaders (99J122).
文摘Objective To investigate the aberrant expressions of TNF-α and TGF-β1 in peripheral blood mononuclear cells （PBMCs） and placental tissues in patients with early spontaneous abortion Methods Using the technique of semi-quantitative reverse transcript-polymerase chain reaction （RT-PCR）, TNF-α mRNA and TGF-β1 mRNA in PBMCs were measured in spontaneous abortion group （30 cases）, normal pregnancy group （25 cases） and nonpregnant group （25 cases）. The expressive intension of TNF-α protein and TGF-β1 protein in placental tissues was also identified by immunohistochemistry. Results Both levels of TNF-α mRNA and TGF-β1 mRNA expressed in PBMCs were significantly different between the three groups respectively （P〈0.05）. Levels of TNF-α in syncytiotrophoblastic and cytotrophoblastic cells of the two aborted groups were substantially higher than those of the nonregnant group （P〈0.01）, but the levels of TGF-β1 in syncytiotrophoblastic cells of the two aborted groups were markedly lower than those of the non-pregnant group （P〈0.01）. Conclusion There is potential relation between TGF-β at the fetomaternal interface and spontaneous abortion. TGF-β1 may contribute to the maintenance of pregnancy, and low-level expression of TGF-β1 may be associated with pregnancy failure.
文摘传统主元分析(Principal component analysis,PCA)方法因忽视量纲对系统的影响,从而使选取的主元难以具有代表性;而在进行量纲标准化后,又因得到的特征值常常是近似相等的而无法进行有效的主元提取.针对这一主要问题,本文通过引入相对化变换(Relative transform,RT)、相对主元(Relative principal components,RPCs)和分布"均匀"等概念,建立起一种相对主元分析(Relative principal component analysis,RPCA)的新方法.该方法首先对系统各分量进行量纲标准化;其次再根据系统的先验信息分析和确定各分量的重要程度;然后在系统能量守恒的准则下,赋以系统各分量相应的权值;最后利用已建立起的相对主元模型,对系统实施RPCA.同时运用数值例子,开展了RPCA在数据压缩和系统故障诊断中的应用研究.理论分析和仿真实验均表明,采用RPCA方法选取出的主元更具代表性和显著几何意义,加之选取主元的灵活性,将使新方法具有更广泛的应用前景。
