自血光化学疗法对中晚期肿瘤放疗增敏的研究--104例的研究

目的将光量子疗法（Ｕｌｔｒａｖｉｏｌｅｔｂｌｏｏｄｉｒｒａｄｉｔｉｏｎ,ＵＢＩ）与光动力疗法（Ｐｈｏｔｏｄｙｎａｍｉｃｔｈｅｒａｐｙ,ＰＤＴ）的理论相结合,利用ＵＢＩ的氧效应ＰＤＴ的光敏作用,探讨其对肿瘤放疗的增敏作用。方法在体外将患者自体抗凝血（２００～３００ＭＬ）加入血卟啉衍生物（Ｈｅｍｏｔｏｐｏｒｐｈｙｒｉｎｄｅｒｉｖａｔｉｖｅ,ＨＰＤ）,经紫外线照射后,回输给病人。回输给２４小时后,行６０钴或６ＭＶＸ线常规分割放疗,治疗中晚期恶性肿瘤１０４例。结果增敏组ＣＲ４６．２％（４８／１０４）,ＰＲ４１．３％（４３／１０４）,ＣＲ＋ＰＲ８７．５％（９１／１０４）,１,２,３年生存率分别为６２．２％,１９．２％,１４．８％。肺癌增敏组与对照组的ＣＲ＋ＰＲ分别为８１．８％（５４／６６）及４４．７％（３０／６７）。子宫颈癌增敏组ＣＲ１００％（１３／１３）,对照组ＣＲ６０％（６／１３）。对照组ＣＲ６０％（６／１３）。结论自血光化学疗法可提高中晚期肿瘤对放疗的敏感性,对肺癌、子宫颈癌的放疗增敏作用尤为显著。     

自血光化学（光动力）疗法放疗增敏治疗中晚期恶性肿瘤200例的研究

利用光量子疗法（ＵＢＩ）的氧效应及促进机体免疫功能、改善微循环的作用,同时借助血卟啉衍生物（ＨＰＤ）的放疗增敏的效应,二者结合与放射治疗综合应用治疗中晚期恶性肿瘤患者２００例。方法是从患者静脉采血２００～３００ｍｌ加入ＨＰＤ１００～１５０ｍｇ（ＨＰＤ...     

膀胱癌中内皮素作用的研究

应用Ｎｏｒｔｈｅｒｎｂｌｏｔ杂交技术及放射免疫方法对人膀胱移行细胞癌组织中内皮素－１ｍＲＮＡ及内皮素活性分子的表达水平进行检测,研究了佛波酯对人膀胱移行细胞癌ＢＩＵ－８７细胞中内皮素活性分子表达水平的调节作用,并探讨了内皮素抗血清及内皮素Ａ型受体竞争性拮抗剂ＢＱ_（１２３）对ＢＩＵ－８７细胞ＤＮＡ合成的影响。结果表明,人膀胱移行细胞癌组织中内皮素－ｌｍＲＮＡ及内皮素活性分子的表达水平明显高于相应的癌旁组织,佛波酯可促进ＢＩＵ－８７细胞合成和分泌内皮素,内皮素抗血清及ＢＱ_（１２３）可抑制ＢＩＵ－８７细胞合成ＤＮＡ。提示内皮素在膀胱癌发展及ＢＩＵ－８７细胞恶性增殖中可能起重要作用。     

沙利度胺联合放疗对宫颈癌裸鼠移植瘤的VEGF、bFGF、TNF-α表达的影响

目的:分析沙利度胺联合放疗对宫颈癌裸鼠移植瘤的VEGF、bFGF、TNF-α表达的影响,探究其增敏作用的机制。方法:选择24只裸鼠,均在左侧腋部皮下注射0.2 m L Hela细胞悬浊液进行造模。将造模成功的裸鼠随机分成对照组(Control group,CG)、沙利度胺组(Thalidomide group,TG)、放疗组(Radiotherapy group,RG)和联合治疗组(TG+RG),每组6只,分别给予羧甲基纤维素钠、沙利度胺溶液、放疗、沙利度胺溶液+放疗处理。治疗14 d后,通过检测裸鼠肿瘤体积、肿瘤抑制率、肿瘤延迟生长时间(Delayed tumor growth time,TGD)、放射增敏比(Radiosensitization ratio,SER)、肿瘤细胞的坏死程度、微血管密度、血清中血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)及肿瘤组织中VEGF、bFGF、肿瘤坏死因子-α(TNF-α)分析沙利度胺联合放疗对宫颈癌裸鼠移植瘤的放疗增敏作用及其机制。结果:治疗14 d后,与对照组相比,沙利度胺组、放疗组和联合治疗组裸鼠肿瘤体积均明显变慢,肿瘤抑制率均显著增高,TGD均显著均明显延长,坏死细胞个数均明显增多,微血管密度、血清和肿瘤组织中VEGF、bFGF、TNF-α表达量均明显降低(P0.05);与沙利度胺组和放疗组相比,联合治疗组裸鼠瘤重均明显变慢,肿瘤抑制率均显著增高,TGD均显著均明显延长,坏死细胞个数均明显增多,微血管密度、血清和肿瘤组中VEGF、bFGF、TNF-α表达量均明显降低(均P0.05);放射增敏比(SER)大于1。结论:沙利度胺可显著增强放疗对宫颈癌裸鼠移植瘤的效果,可能与其显著降低VEGF、bFGF、TNF-α的表达有关。     

TGFa反义寡聚核苷酸对膀胱癌BIU87细胞株的作用

报道了２３碱基组成的ＴＧＦａ反义寡聚核苷酸和对照寡聚核苷酸对人膀胱移行细胞癌ＢＩＵ８７细胞增殖及其ＴＧＦａｍＲＮＡ表达的表用,结果表明：ＴＧＦａ反义寡聚核苷酸抑制抑体外培养的ＢＩＵ８７细胞的增殖,ＤＮＡ合成和ＴＧＦａｍＲＮＡ的表达,进一步证明了ＴＧＦａ在ＢＩＵ８７细胞恶性增殖中的重要作用。     

单克隆抗体ELISA测定UVB诱导的DNA损伤(英文)

本文采用单克隆抗体酶联免疫吸附分析法测定了ＵＶＢ诱导ＤＮＡ产生的ＣＰＤ和６４ＰＰ。经０．５ｍＷ／ｃｍ２ＵＶＢ处理１５ｍｉｎ的小牛胸腺和鲱鱼精ＤＮＡ,ＣＰＤ和６４ＰＰ含量显著增加,而未经ＵＶＢ处理的对照ＤＮＡ则没有二聚体形成。     

单克隆抗体ELISA测定UVB诱导的DNA损伤

本文采用单克隆抗体酶联免疫吸附分析法测定了ＵＶ－Ｂ诱导ＤＮＡ产生的ＣＰＤ和６－４ＰＰ。经０．５ｍＷ／ｃｍ＾２ＵＶ－Ｂ处理１５ｍｉｎ的小牛胸物鲱鱼精ＤＮＡ,ＣＰＤ和６－４ＰＰ含量显著增加,而未经ＵＶ－Ｂ处理的对照ＤＮＡ则没有二聚体形成。     

自血光化学疗法对放疗肿瘤患者细胞免疫功能的影响

本文通过对肿瘤患者Ｅ玫瑰花环及淋巴细胞转化率的检测,探讨自血光化学放射敏疗法对机体免疫功能的影响。结果表明：单纯放疗组Ｅ玫瑰花环形成及淋巴细胞转化率较放职前明显下降,而自血光化学放疗增敏组治疗前后上述免疫功能无明显改变,表明自血光化学疗法对肿瘤放疗患者的细胞免疫功能具有保护作用。     

姜黄素对晚期宫颈癌患者Bcl-xL、Bcl-2、EphA2及EphrinA1表达的影响

目的:探究姜黄素对宫颈癌晚期放疗患者Bcl-x L、Bcl-2、Eph A2与Ephrin A1表达的影响。方法:收集160例晚期宫颈癌放疗患者的宫颈癌石蜡标本,分为姜黄素组与对照组,均给予放射治疗,姜黄素组放疗期间加服姜黄素片剂,采用免疫组化等实验方法,观察两组病例标本Bcl-x L、Bcl-2、Eph A2与Ephrin A1的表达情况,比较两组病理标本化疗前后的凋亡细胞指数(AI)与微血管密度(MVD)。结果:姜黄素组的Bcl-x L、Bcl-2、Eph A2与Ephrin A1的表达阳性率分别为8.3%、43.33%、46.67%、15.0%,均显著低于对照组(P0.05);放疗后姜黄素组的AI显著高于对照组(P0.05),MVD显著低于对照组(P0.05)。结论:姜黄素可抑制Bcl-x L、Bcl-2的表达以促进肿瘤细胞凋亡,同时降低Eph A2与Ephrin A1的表达以减少肿瘤微血管形成,有显著的抗肿瘤与放疗增敏作用。     

肿瘤放射治疗增敏药物的研究与应用

本书由郑秀龙,金一尊等九位多年从事放射生物学研究、教学及临床肿瘤放射治疗工作者撰写.该书不仅较系统地介绍了放射增敏作用的基本概念、作用机制及临床应用,还着重介绍了国内外放射增敏剂的最新成就和最新方法以及放射增敏剂的发展趋势.该书是     

光量子加血卟啉疗法对大鼠肿瘤放射增敏作用的对比研究

本文应用透射电子显微镜技术观察了应用血卟啉加光量子疗法后,再施以放射治疗,大鼠皮下移植性肿瘤Ｗ２５６细胞超微结构的变化,结果表明,与单纯充氧放疗组的肿瘤细胞相比,经过血卟啉,光量子疗法及同时应用血卟啉和光量子疗法后再施放疗组大鼠组织周围有明显增多的炎性细胞浸润及淋巴细胞浸润;肿瘤细胞可见有不同程度的核改变及粗面内质脱颗闰,扩张：线粒体肿胀,嵴及膜的断裂,基质丢失,应用血卟啉及血卟啉加光量子同时应用     

光量子和血卟啉的放疗增敏作用与表皮生长因子关系的研究

本文采用放射免疫技术检测了充氧及充氧并用光量子或血卟啉和光量子与血卟啉联合治疗后进行局部放射治疗皮下Ｗ２５６移植瘤大鼠的血浆、瘤组织和颌下腺组织内表皮生长因子（ＥＧＦ）的含量。结果表明,与单纯充氧后放射治疗组大鼠比较：（１）光量子治疗组、血卟啉治疗组和光量子与血啉卟联合治疗组大鼠血浆ＥＧＦ含量显著升高（Ｐ〈０．０５）;（２）光量子治疗组、血卟啉治疗组和光量子与血卟啉联合治疗组大鼠组织与颌下腺中ＥＧ     

Evaluation of the stress-inducible production of choline oxidase in transgenic rice as a strategy for producing the stress-protectant glycine betaine

Glycine betaine (GB) is a compatible solute that is also capable of stabilizing the structure and function of macromolecules. Several GB-producing transgenic rice lines were generated in which the Arthrobacter pascens choline oxidase (COX) gene, fused to a chloroplast targeting sequence (TP) was expressed under the control of an ABA-inducible promoter (SIP; stress-inducible promoter) or a ubiquitin (UBI) gene promoter that is considered to be constitutive. This comparison led to interesting observations that suggest complex regulation with respect to GB synthesis and plant growth response under stress. In spite of the use of the well-studied stress-inducible promoter, the highest level of GB accumulation (up to 2.60 micromol g(-1) DW) in the SIP lines grown under saline conditions was not as high as in the UBI lines (up to 3.12 micromol g(-1) DW). Therefore, the use of an ABA-inducible promoter was not more beneficial for de novo production of GB. Interestingly, saline growth conditions enhanced GB accumulation by up to 89% in the SIP lines, whereas up to 44% increase was seen in a UBI line. In all these cases the GB levels were many-fold below the range reported for plant species that produce GB naturally. In spite of lower GB concentrations, statistically greater levels of stress tolerance were found in SIP lines than in UBI lines, suggesting that the stress protection observed in SIP plants cannot be totally explained by the increase in the GB content.     

Induction of a polyubiquitin gene promoter by dehydration stresses in transformed rice cells

The expression of the maize polyubiquitin gene promoter UBI1 in rice cells has been used to study the involvement of ubiquitin in cell protection responses to dehydration caused by osmotic, saline or freezing stress. The effect of these stresses on UBI1 activity was investigated by the use of stably transformed rice calli (UBI1:GUS), as well as by transient expression experiments performed with cell lines with high or low tolerance to each type of stress. The theoretical analysis of the UBI1 promoter shows several putative stress-regulated boxes that could account for the stress-related UBI1 induction pattern described in this work. We suggest that the study of the differential UBI1 promoter-driven expression in rice cell lines with different level of tolerance to stress might be useful to elucidate complex signal transduction pathways in response to dehydration stresses in monocots.     

UBI4, the polyubiquitin gene of Saccharomyces cerevisiae, is a heat shock gene that is also subject to catabolite derepression control

Carbon and nitrogen regulation of UBI4, the stress-inducible polyubiquitin gene of Saccharomyces cerevisiae, was investigated using a UBI4 promoter-LacZ fusion gene (UBI4-LacZ). Expression of this gene in cells grown on different media indicated that the UBI4 promoter is more active during growth on respiratory than on fermentable carbon sources but is not subject to appreciable control by nitrogen catabolite repression. UBI4-LacZ expression was virtually identical in cells having constitutively high (ras2, sra1-13) or constitutively low (ras2) levels of cyclic AMP-dependent protein kinase activity, indicating that this kinase does not exert a major influence on UBI4 expression. Catabolite derepression control of the UBI4 promoter was confirmed by measurements of UBI4-LacZ expression in hap mutant and wild-type strains before and after transfer from glucose to lactate. Mutagenesis of the perfect consensus for HAP2/3/4 complex binding at position ?542 resulted in considerable reduction of UBI4 promoter derepression with respiratory adaptation in HAP wild-type cells and abolished the reduced UBI4-LacZ derepression normally seen when aerobic cultures of the hap1 mutant are transferred from glucose to lactate. This HAP2/3/4 binding site is therefore a major element contributing to catabolite derepression of the UBI4 promoter, although data obtained with hap1 mutant cells indicated that HAP1 also contributes to this derepression. The HAP2/3/4 and HAP1 systems are normally found to activate genes for mitochondrial (respiratory) functions. Their involvement in mediating higher activity of the UBI4 promoter during respiratory growth may reflect the contribution of UBI4 expression to tolerance of oxidative stress.     

UBI4, the polyubiquitin gene of Saccharomyces cerevisiae , is a heat shock gene that is also subject to catabolite derepression control

 Carbon and nitrogen regulation of UBI4, the stress-inducible polyubiquitin gene of Saccharomyces cerevisiae, was investigated using a UBI4 promoter-LacZ fusion gene (UBI4-LacZ). Expression of this gene in cells grown on different media indicated that the UBI4 promoter is more active during growth on respiratory than on fermentable carbon sources but is not subject to appreciable control by nitrogen catabolite repression. UBI4-LacZ expression was virtually identical in cells having constitutively high (ras2, sra1-13) or constitutively low (ras2) levels of cyclic AMP-dependent protein kinase activity, indicating that this kinase does not exert a major influence on UBI4 expression. Catabolite derepression control of the UBI4 promoter was confirmed by measurements of UBI4-LacZ expression in hap mutant and wild-type strains before and after transfer from glucose to lactate. Mutagenesis of the perfect consensus for HAP2/3/4 complex binding at position −542 resulted in considerable reduction of UBI4 promoter derepression with respiratory adaptation in HAP wild-type cells and abolished the reduced UBI4-LacZ derepression normally seen when aerobic cultures of the hap1 mutant are transferred from glucose to lactate. This HAP2/3/4 binding site is therefore a major element contributing to catabolite derepression of the UBI4 promoter, although data obtained with hap1 mutant cells indicated that HAP1 also contributes to this derepression. The HAP2/3/4 and HAP1 systems are normally found to activate genes for mitochondrial (respiratory) functions. Their involvement in mediating higher activity of the UBI4 promoter during respiratory growth may reflect the contribution of UBI4 expression to tolerance of oxidative stress. Received: 3 June 1996 / Accepted: 20 August 1996     

CLONING AND EXPRESSION OF SPODOPTERA LITURA UBIQUITIN GENE

Abstract Ubiquitin (UBI) plays a very important role in regulated non-lysosomal ATP dependent protein degradation. In the present work, the coding sequence of Spodoptera litura UBI gene was isolated (GenBank Accession No. AF436066). The length of this ORF is 228bp, encoding a protein with Mr of 8.56 kD and isoelectric point of 6.56. Multiple sequence alignment indicated that S. litura UBI is very similar to the homologous proteins of other eukaryotic species and it has 84% identity with S. litura nucleopolyhedrovirus (SpltMNPV) UBI at amino acid level. RT-PCR analysis showed that S. litura UBI gene is ubiquitously expressed in larva tissues which are susceptible to SpltMNPV infection. By constructing E. coli expression vector, S. litura UBI was highly expressed and the recombinant protein was purified using Ni-NTA resin column, and currently further study on the function of S. litura UBI in SpltMNPV infection is underway.     

斜纹夜蛾泛素基因的克隆及表达

泛素介导的蛋白质降解途径对脑内蛋白的选择性降解起着重要作用。设计一对简并引物,从斜纹夜娥(Spodoptera litura)细胞中克隆了泛素基因的编码区,CenBank登录号AF436066。序列分析表明,该编码区的长度为228bp,编码由76个氨基酸组成的、分子质量为8．56kD的蛋白,其等电点为6．56。同源性比较发现,斜纹夜峨泛素基因不仅与其它真核生物的泛素基因在氨基酸水平上具有96％以上的相似性,而且与斜纹夜蛾核多角体病毒(SpltMNPV)泛素基因的同源性为84％。RT—PCR分析发现,泛素基因在所检测的斜纹夜蛾幼虫多种组织,尤其是脂肪体中均有表达。采用构建的原核表达载体pQEUB,在大肠杆菌M15中诱导并高效表达出了带有His—tag的重组融合蛋白,薄层扫描分析得知靶蛋白约占总蛋白的37％。利用Ni—NTA亲和层析胶纯化得到重组融合蛋白,经SDS—PAGE鉴定为单一区带,为进一步研究S．litura泛素在SpltMNPV感染中的作用打下了基础。