人良性增生前列腺及上皮和间质细胞的鸟氨酸脱羧酶活性

以比色法测定了正常人（８例）和良性前列腺增生症病人（２１例）前列腺组织的全匀浆、上皮和间质细胞中的鸟氨酸脱羧酶（ＯＤＣ）的活性。结果显示：正常组织的上皮部分ＯＤＣ活性高于间质部分，而增生组织相应的两部分ＯＤＣ活性无差异；增生组织的间质部分酶活性是正常组织该部分酶活性的２．５倍，两组间上皮部分的酶活性无差异；增生组织全匀浆的酶活性显著高于正常组织。结果表明前列腺组织中ＯＤＣ活性升高与良性前列腺增生症     

良性增生前列腺组织及上皮和间质细胞中r—GT活性及GSH含量…

本文报道测定正常人和良性腺增生症（ＢＰＨ）患者前列腺组织匀浆、上皮及间质细胞中ｒ－谷氨酰转肽酶（ｒ－ＧＴ）活性及其底物还原型谷脱甘肽（ＧＳＨ）的含量。结果显示：①ＢＰＨ组织匀浆、上皮及间质细胞中ｒ－ＧＴ活性显著升高，分别为正常前列腺相应组分的４．５、３．２和４．１倍，而ＧＳＨ含量则明显降低；②正常及ＢＰＨ组织上皮细胞中ｒ－ＧＴ活性均显著高于相应间质细胞，而ＧＳＨ含量则低于间质细胞。提示ｒ－ＧＴ活性     

雄激素相关酶活性与BPH关系的研究

测定良性前列腺增生症(BPH)和正常人前列腺组织及不同细胞中5α-还原酶、鸟氨酸脱羧酶(ODC)及γ-谷氨酰转肽酶(r-GT)活性。结果显示:(1)5α-还原酶主要存在于前列腺间质细胞核中,BPH前列腺间质细胞中5α-还原酶活性约为正常前列腺的2倍;(2)BPH前列腺全匀浆和间质细胞中ODC活性分别为正常组织相应组分的2.4和2.6倍;(3)BPH前列腺全匀浆、上皮和间质细胞中r-GT活性分别为正常前列腺相应组分的4.5倍、3.2倍和4.1倍。提示5α-还原酶、ODC及r-GT活性升高与BPH的发生可能有密切关系。     

口腔粘膜白斑和鳞癌组织中p53和bcl—2蛋白的表达

目的：研究凋亡相关蛋白ｂｃｌ－２、Ｐ５３在口腔粘膜、上皮异常增生和鳞癌组织中的表达及意义。方法：采用免疫组织化学Ｓ－Ｐ检测１０例正常口腔粘膜,１０例单纯性增生上皮、３０例异常增生上皮和３３例鳞癌石蜡包埋组织中ｂｃｌ－２、、ｐ５３的表达。结果：正常青草帮上皮增生组织中未见Ｐ５３阳性表达。异常增生上皮和鳞癌中Ｐ５３阳性率 ３３．３３％和４５．４５％,与正常组和单纯增生组相比有显著怀差异。ｂｃｌ－２在正     

子宫内膜、正常早孕和自然流产蜕膜组织中△Np63的表达

目的观察△Np63蛋白在自然流产蜕膜组织中的表达，探讨△Np63在胚泡植入和早期妊娠维持中的作用。方法收集自然流产的蜕膜27例组织（其中胚胎停止发育23例、习惯性流产1例及不全流产3例），正常早孕蜕膜66例，非妊娠期子宫内膜组织57例（其中增生期33例，分泌期24例），用SP免疫组化染色法观察△Np63的表达。结果△Np63广泛表达于腺上皮和间质细胞中。增生期和分泌期子宫内膜中△Np63主要表达在腺上皮，蜕膜组织中△Np63在间质蜕膜细胞中的有较强的表达。流产组△Np63蛋白阳性率为55．6％，明显低于增生期的90．9％、分泌期的87．5％及正常早孕组的86．4％（P〈0．05）。△Np63高表达率在分泌期组及流产组分别为16．7％和11．2％，明显低于增生期的63．6％和正常早孕组的57．6％（P〈0．01）。结论△Np63可能参与子宫内膜的生理周期变化，并与子宫内膜向早孕蜕膜组织转化及维持早孕关系密切。     

男性后尿道内窥镜解剖学要点及临床应用

在内窥镜下，观察测量了２３５例正常男性成人和３０４例前列腺增生病人的后尿道形态学要点。两组测量结果的平均值，在尿道前列腺上下径（分别３．６ｃｍ及６．８ｃｍ），膀胱颈最高点至精阜距离（分别为２．６ｃｍ，及３．８ｃｍ）均有显著差异。结合后尿道形态学特点，为９３例前列腺增生症患者施行了经尿道前列腺联合部切开术，讨论了术式设计有关的解剖学要点。     

人肝癌及肝硬变中IGF—2及HBxAg表达的对比研究

对６０例肝细胞癌、癌旁肝组织和４７例肝硬变石蜡切片进行ＩＧＦ－２和ＨＢｘＡｇ免疫组化染色。结果表明，在ＨＢｘＡｇ阳性的肝癌和肝硬变中ＩＧＦ－２阳性率分别为１００％（３２／３２例）和９４．６％（３５／３７例），而ＨＢｘＡｇ阴性组分别为８２．１％（２３／２８例）和６０％（６／１０例）。ＩＧＦ－２在肝癌中的阳性强度明显高于癌旁肝和肝硬变。ＩＧＦ－２的分布形态有：（１）胞浆包涵体样、（２）全胞浆、（３）核     

失血性休克条件下磷脂酶A2（PLA2）和细胞膜脂流动性的改变

本研究采用家兔失血性休克模型，测定失血性休克条件下血浆和组织ＰＬＡ＿２活性的变化和红细胞膜流动性的改变。结果表明失血性休克前和休克后１、２、５小时血浆ＰＬＡ＿２分别为４６．９±１２．２ｍＭ／Ｌ、９４．０±５０．０、１１９．２±６２．４、１７４．５±８５．３ｍＭ／Ｌ，休克后血浆ＰＬＡ＿２比伤前显著升高（Ｐ＜０．０１），为伤前的２．０、２．５、３．７倍。失血性休克５ｈ后肝、心、肺、肠粘膜中ＰＬＡ＿２含量为假手术组的２．４７５、３．４７５、３．３３９、４．６５７倍，显著高于假手术组。同时失血性休克时红细胞膜流动性降低，膜脂分子排列有序性升高，细胞膜荧光偏振度升高，红细胞膜脂区微粘度升高。血浆ＰＬＡ＿２的升高和细胞膜流动性改变呈显著正相关（ｒ＞０．９３）。结果说明：失血性休克时ＰＬＡ＿２水平升高可能是血液动力学衰竭和膜损伤的重要原因，ＰＬＡ＿２是失血性休克的一个关键酶和重要的体液因子。     

p53基因突变与食管癌生物学行为的关系

目的为了探讨ｐ５３基因突变与食管癌生物学行为及预后的关系。方法应用ＰＣＲ－ＳＳＣＰ结合ＤＮＡ直接银染测序，对３０例散发性食管癌组织ｐ５３基因第５～８外显子进行检测。结果检出１１例阳性，突变检出率为３６．７％。９例为点突变，其中错义突变４例、无义突变２例、同义突变３例，其余２例为碱基插入和缺失导致的移码突变。统计学分析显示：中低分化食管癌的ｐ５３突变率为５６．３％，高分化组为１４．３％，两组相比有非常显著性差异（Ｐ＝０．０２５）；癌组织浸润累及食管壁全层的ｐ５３突变率５２．６％显著高于未累及全层组９．１％（Ｐ＝０．０２４）；有淋巴结转移组与无淋巴结转移组相比，ｐ５３突变率分别为６１．５％和１７．６％也具有非常显著性差异（Ｐ＝０．０２４）。结论ｐ５３基因突变与食管癌多种生物学行为如组织分化程度、肿瘤浸润程度及淋巴结转移有明显相关性。因此检测食管癌组织中是否存在ｐ５３基因突变有助于判断食管癌的恶性程度和患者的预后。还讨论了ｐ５３基因的“显性负效应”及同义突变的遗传学效应。     

过敏性哮喘血浆可溶性白介素2受体与嗜酸性细胞、嗜碱性细胞、总IgE关系的研究

３０例过敏性哮喘患者和２４例正常人分别被检测了血浆可溶性白介素２受体（ＳＩＬ－２Ｒ）、ＩｇＥ和全血嗜酸性细胞、嗜碱性细胞值，可观察到哮喘组ＳＩＬ－２Ｒ平均值为２３２０ｐｇ／ｍＬ，明显高于正常组的１７６０．９４ｐｇ／ｍＬ）（Ｐ＜０．０５），表明哮喘组Ｔ细胞呈活化状况。哮喘组的嗜酸性细胞、嗜碱性细胞、ＩｇＥ增高的百分率分别为６３．３３％（１９／３０）、６６．６７％（２０／３０）和３６．６７％（１１／３０），均明显高于正常组的４．１７％（１／２４）．哮喘组中有８例ＳＩＬ－２Ｒ值大于３０００ｐｇ／ｍＬ，其中４例（５０％）嗜酸性细胞、ＩｇＥ２项检测及嗜酸性细胞、ＩｇＥ、嗜碱性细胞３项检测均增高，且病情较重，而在ＳＩＬ－２Ｒ≤３０００ｐｇ／ｍＬ的２２例中，其２项、３项检测同时增高者仅３例（１３．６４％），２组差异显著（Ｐ＜０．０５），提示血浆ＳＩＬ－２Ｒ的检测对研究哮喘的发病机理和反映病情严重状况均有意义。     

丝裂素激活蛋白激酶在良性前列腺增生中的表达及临床意义

目的对比研究丝裂素激活蛋白激酶(MAPKs)在增生与正常前列腺组织中的表达,探讨良性前列腺增生发病的分子生物学机制。方法采用W estern B lot及免疫组织化学方法,检测增生及正常前列腺组织中ERK、JNK和p38MAPK的表达情况。结果与正常前列腺组织相比,增生组织ERK的平均灰度值(wh ite~b lack,0~255)和阳性细胞百分率明显升高,JNK和p38MAPK的检测结果则与之相反。ERK染色主要位于细胞核和细胞质内,在增生组织上皮细胞及基质细胞的染色阳性率均高于正常前列腺组织。结论ERK在良性前列腺增生组织中过表达,从而导致细胞增生指数增加,JNK和p38MAPK低表达导致凋亡减少,促进疾病的发生和发展。     

Changes in ornithine decarboxylase activity and putrescine concentrations after spinal cord compression injury in the rat.

Traumatic spinal cord injury results in direct physical damage to structures and the generation of local factors contributing to secondary pathogenesis. In the present study, we investigated changes in polyamine metabolism after spinal cord compression injury in the rat. This is a stress induced metabolic pathway, of which an activation may indicate both, secondary pathogenesis or induction of neuroprotective response. Ornithine decarboxylase (ODC) activity, the rate limiting step of polyamine synthesis, and levels of the diamine putrescine, the product of ornithine decarboxylase reaction, were analyzed in control (non-laminectomized) animals and at 2 and 4 h after laminectomy or compression injury at the L4 segmental level. ODC activity was significantly increased 4 h after laminectomy in L4 and in adjacent L3 and L5 segments and compression to L4 produced a further increase 4 h after injury as compared with the intact control group. Putrescine levels were likewise significantly elevated to the same extend in the laminectomized and injured cord as compared with the intact control group. These findings demonstrate increased ODC and putrescine levels in the laminectomized and traumatized spinal cord and suggest that laminectomy may be an important 'priming event' that contributes to secondary injury after spinal cord compression injury.     

Effects of luminal stimuli on polyamine metabolism in the small intestine of the rat: the role of enteric nerves

The aim of this study was to investigate to what extent polyamine metabolism in the small intestine of the rat is controlled by the enteric nervous system. Polyamine metabolism was followed by measuring the activity of ornithine decarboxylase (ODC) and in some instances also the content of polyamines (putrescine, spermidine and spermine). ODC activity in the intestine was increased when intraluminal pressure was increased and 3 h after placing cholera toxin in the intestinal lumen. Cholera toxin also increased the tissue putrescine content. Atropine or hexamethonium given i.v. did not influence the evoked changes of ODC activity. The pressure induced changes were not decreased by placing lidocaine on the serosal surface. On the other hand, the ODC activity of control segments were decreased by hexamethonium or atropine. The presence of glucose in the intestinal perfusate did not augment tissue ODC activity, neither did the heat stable enterotoxin from Escherichia coli (STa). It is concluded that the effect on polyamine metabolism evoked by luminal pressure or cholera toxin seems not to be mediated via nerves, while nerves seem to influence ODC activity during control conditions. The experiments with enterotoxins suggest that cAMP is the intracellular second messenger controlling intestinal ODC activity.     

Interleukin-8 Is a Paracrine Inducer of Fibroblast Growth Factor 2, a Stromal and Epithelial Growth Factor in Benign Prostatic Hyperplasia

Benign prostatic hyperplasia (BPH) is an extremely common disease of older men in which there is benign overgrowth of the prostatic transition zone, leading to obstruction of urine outflow. Fibroblast growth factor (FGF) 2, a potent growth factor for prostatic stromal and epithelial cells, is increased twofold in BPH and its concentration is correlated with stromal proliferation in this condition. Immunohistochemistry of normal and hyperplastic prostate revealed that FGF2-expressing stromal cells were present in higher numbers near the epithelial acini, implying that epithelial cells may express a factor that induces FGF2 expression by stromal cells. Conditioned medium from primary cultures of prostatic epithelial cells was capable of inducing increased expression of FGF2 by primary stromal cultures. Blocking experiments with neutralizing anti-interleukin (IL)-8 antibodies and pretreatment with lipopolysaccharide, which down-regulates the IL-8 receptor, show that this inducing activity is because of the presence of IL-8 in the epithelial-conditioned medium. Analysis of normal prostatic peripheral zone and BPH tissue by enzyme-linked immunosorbent assay reveals that IL-8 is present at increased levels in hyperplastic prostate. Therefore IL-8 produced by prostatic epithelial cells can induce FGF2, a potent stromal and epithelial growth factor, and in this manner promote the abnormal proliferation of the prostatic transition zone that is critical in the pathogenesis of BPH.     

Regulation of Polyamine Synthesis and Transport by Fibroblast Growth Factor in Aortic Smooth Muscle Cells

Abstract

Basic-FGF (FGF2) is implicated as a regulator of smooth muscle cell proliferation that develops after arterial injury. Polyamines are essential for cell growth and differentiation and may mediate some of the FGF2-elicited responses. To examine this possibility, the effect of FGF2 on polyamine synthesis and uptake was tested on rat arterial smooth muscle cells. Exposure of cells to FGF2 for 24 and 48 h resulted in increased intracellular polyamine content. Ornithine decarboxylase (ODC) activity increased in FGF2-treated cells after 6 h of treatment, whereas no increases were detected in ODC mRNA steady-state levels. Basic-FGF increased maximal polyamine transport rate without changes in Km. Treatment with actinomycin D decreased polyamine transport. The effect of cyclohexamide on polyamine uptake was dose dependent. These studies indicate that treatment of vascular smooth muscle cells with FGF2 results in increases in intracellular polyamine content, polyamine synthetic activity, and polyamine transport.     

运动训练对老龄大鼠心肌多胺代谢的影响

目的:通过观察运动训练对老龄大鼠心肌多胺代谢、心肌抗炎及抗氧化能力的影响,探讨多胺代谢在运动训练延缓心肌衰老中的作用。方法:实验分为3组:老年运动组(Old+Ex),18月龄Wistar大鼠梯度跑台运动6周;老年组(Old),与Old+Ex组月龄相同的Wistar大鼠;青年组(Young),3月龄Wistar大鼠。高效液相色谱测定心肌组织中多胺(腐胺、精脒和精胺)含量;[¹⁴C]标记液闪计数方法测定心肌组织中多胺合成限速酶鸟氨酸脱羧酶(ODC)与多胺分解限速酶精脒/精胺乙酰转移酶(SSAT)活性;比色法检测心肌组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;酶联免疫吸附(ELISA)法检测心肌组织中肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)含量;超声心动图记录大鼠心脏功能;透射电镜观察心肌组织超微结构变化。结果:与Young组比,Old组大鼠心肌组织中ODC活性下降, SSAT活性升高, 精脒、精胺及总多胺池水平显著降低;心肌SOD活性下降,MDA、TNF-α和IL-1β水平升高(P<0.05);超声心动图显示,Old组大鼠左心室收缩末期直径(LVESD)与舒张末期直径(LVEDD)均明显增大,左心室射血分数(LVEF)与缩短分数(LVFS)降低(与Young组比,P<0.01)。Old+Ex组大鼠心肌组织中ODC活性增加,SSAT活性下降,精脒及总多胺池水平明显增加;心肌SOD活性升高,MDA、TNF-α和IL-1β水平均显著降低(与Old组比,P<0.05);左室功能有明显改善。超微结构观察可见Old组心肌肌丝排列不整齐,可见大量脂褐素颗粒沉积,线粒体基质疏松;Old+Ex组心肌肌节结构清晰,线粒体基质致密,嵴排列整齐。结论:运动训练通过上调多胺合成代谢、抑制其分解代谢对抗衰老引起的心肌多胺水平降低;运动训练可提高老龄心肌抗炎、抗氧化能力。维持老龄心肌多胺池在适当水平可能是运动延缓心肌衰老的部分机制之一。     

前列腺病变组织中细胞凋亡与bcl-2、bax、PCNA表达

目的　研究前列腺良、恶性病变组织中细胞增殖与凋亡及其与相关蛋白表达意义。方法　采用原位细胞凋亡标记技术 (TUNEL)及免疫组织化学ABC法对 36例前列腺癌 (PCa)、2 0例前列腺增生 (BPH)和 11例正常前列腺 (NP)组织石蜡切片bcl 2、bax、PCNA蛋白及细胞凋亡检测。结果　前列腺癌细胞凋亡指数 (AI)高于BPH和NP(P <0 0 1) ,bcl 2蛋白阳性表达者AI低于阴性者 (P <0 0 1) ,bax蛋白阳性表达者AI高于阴性者 (P <0 0 5 ) ;前列腺癌和BPH的bcl 2和PCNA蛋白阳性表达率高于NP(P <0 0 5 ) ,并随着肿瘤分级增高而增高 ;NP、BPH和前列腺癌的bax蛋白阳性表达率差异无显著性。前列腺癌细胞增殖指数 (PI)明显高于BPH(P <0 0 1) ,BPH细胞PI较NP明显增高 ,但细胞AI却显著下降。结论　细胞增殖与细胞凋亡的增加在PCa的发生和发展中起到了重要作用。在BPH的形成过程中前列腺组织细胞增殖增加而细胞凋亡减少 ,其中bcl 2和bax在细胞凋亡调节中起重要作用。     

Interleukin-1alpha is a paracrine inducer of FGF7, a key epithelial growth factor in benign prostatic hyperplasia

Benign prostatic hyperplasia (BPH) is an extremely common disease of older men in which there is benign overgrowth of the prostatic transition zone, leading to obstruction of urine outflow. FGF7, a potent growth factor for prostatic epithelial cells, is increased by threefold in BPH and is correlated with increased epithelial proliferation in this condition. Immunohistochemistry of normal and hyperplastic prostate revealed that FGF7-expressing fibroblastic cells were present in higher numbers near the epithelial acini, implying that epithelial cells may express a factor that induces FGF7 expression by stromal cells. Conditioned medium (CM) from primary cultures of prostatic epithelial cells was capable of inducing a two- to sixfold increase in the expression of FGF7 by primary stromal cultures. Blocking experiments with neutralizing anti-interleukin-1alpha (Il-1alpha) antibodies and IL-1Ra, an Il-1alpha receptor antagonist, show that this inducing activity was due to the presence of Il-1alpha in the epithelial CM. Analysis of normal prostatic peripheral zone and BPH tissue by enzyme-linked immunoabsorption assay reveal that Il-1alpha is present at increased levels in hyperplastic prostate and that levels of Il-1alpha correlate strongly with tissue FGF7 concentration in BPH. Therefore Il-1alpha is produced by prostatic epithelial cells and can induce FGF7, a potent epithelial growth factor, which can in turn lead to further epithelial growth and increased Il-1alpha secretion, thus establishing a double paracrine loop that is functionally equivalent to an autocrine growth loop. This double paracrine loop may play a key role in the abnormal proliferation of the transition zone, which is critical to the pathogenesis of BPH.     

Subacute combined degeneration and induction of ornithine decarboxylase in spinal cords of totally gastrectomized rats

Totally gastrectomized rats have been used to induce a spongy demyelination in the white matter of the spinal cord (SC) which is strongly reminiscent of that observed in subacute combined degeneration of human SC. Totally gastrectomized rats are deprived of intrinsic factor and thereafter become deficient in cobalamin. Morphologically, the spongy demyelination of the white matter of the rat SC, was evident 2 months after total gastrectomy. Biochemically, we investigated the hypothesis that polyamine biosynthesis might be deranged in the rat SC with experimental subacute combined degeneration, since polyamines are well known to be bound to myelin in the mammalian central nervous system. We measured the levels of both the polyamine biosynthetic decarboxylases, L-ornithine decarboxylase (ODC) and S-adenosyl-L-methionine decarboxylase, the key points in the polyamine biosynthetic pathway, in these SC. There was a sharp increase in ODC activity in SC 2 months after total gastrectomy, without significant changes in S-adenosyl-L-methionine decarboxylase activity. The increase in ODC activity seems to be organ-specific and was not due to a proliferation of neuroglial cells. Interestingly enough, the same morphologic and biochemical features found in SC of 2-month-totally-gastrectomized rats were present also in SC of newborn rats, which indeed showed incomplete myelination, vacuolated appearance, and an ODC activity level higher than that of adult SC. Therefore, total gastrectomy seems to induce a type of regression in the SC of totally gastrectomized rats toward neonatal life, at least in terms of the degree of myelination and of ODC activity level. Biochemically, no changes in ODC activity were observed in SC of rats fed a cobalamin-deficient diet for 3 months. Morphologically, only a proliferation of neuroglial cells with a moderate demyelination was observed in SC of these rats maintained on a cobalamin-deficient diet for 3 months.     

Polyamines modulate carcinogen-induced mutagenesis in vivo

Elevated polyamine levels as a consequence of targeted overexpression of ornithine decarboxylase (ODC) to murine skin enhance susceptibility to tumorigenesis in this tissue. A possible mechanism for the enhanced susceptibility phenotype is an increased sensitivity of tissues with elevated polyamine levels to the mutagenic action of carcinogens. To test this hypothesis, a transgenic mouse model containing the Big Blue transgene and also expressing a K6/ODC transgene was developed. Incorporation of the K6/ODC transgene into the Big Blue model did not affect the spontaneous lacI mutant frequency in either skin or epidermis of the double-transgenic mice. After skin treatment with single doses of either 7,12-dimethylbenz[a]anthracene or N-methyl-N'-nitro-N-nitrosoguanidine, however, the mutant frequency was significantly increased in the skin of double-transgenic Big Blue;K6/ODC mice compared to Big Blue controls. The increases in mutant frequency were clearly due to ODC transgene activity, since treatment of mice with the ODC inhibitor, alpha-difluoromethylornithine, completely abolished the difference in mutant frequencies between double-transgenic and Big Blue mice. These results demonstrate that intracellular polyamine levels modulate mutation induction following carcinogen exposure.