恒河猴黄体中tPA,uPA和PAI-1的鉴别与其功能的研究

在大鼠和恒河猴卵巢颗粒细胞(GC)存在组织型(tPA)和尿激酶型(uPA)纤溶酶原激活因子(PA)和一种PA的抑制因子(PAI-1).在排卵、排精和子宫内膜的周期性变化中tPA和PAI-1基因在这些组织的同类细胞或不同类型细胞间的协调表达起重要作用.因为恒河猴GC和膜-间质细胞(TC)在促性腺激素作用下都能产生tPA,uPA和PAI-1,而排卵后GC和TC转化为黄体细胞(LC),后者分泌孕酮,维持妊娠.本研究目的:(1)探讨LC是

人和恒河猴胎盘基盘分泌tPA和PAI-1部位的鉴定

tPA和PAI-1集中于恒河猴子宫肌蜕膜层与分离蜕膜交界层中。子宫肌蜕膜组织以及肌层细胞中皆为阴性反应。在蜕膜组织血管周围有明显PAI-1的分布,证实了tPA和PAI-1 在母体与胎儿分离机制中起重要作用。

猕猴精浆纤溶酶原激活因子的来源及在精子获能中的作用

猕猴附睾、前列腺和精囊均表达tPA、uPA和PAI-1 mRNAs。加入uPA能维持精子的活力,使精子产生超激活运动,诱导顶体反应的发生,并使精子获得激活卵子的能力。这说明猕猴精浆PA除来源于睾丸外,可能主要来源于附睾及附性腺;在体外,uPA,而不是tPA,可能诱导精子获能。

纤溶酶原激活因子与猕猴精子运动力的关系及其在附睾和附性腺中的表达调节

原位杂交的结果表明11酸睾酮诱导少精子症和弱精子症,tPA mRNA的表达在附睾头、精囊及前列腺减少,而在附睾体升高,附睾尾表达基本无变化;uPA mRNA的表达在附睾头、附睾体、前列腺减少,而在精囊升高,附睾尾表达基本无变化;PAI-1 mRNA的表达在附睾头、附睾体、精囊下降,而在前列腺升高,附睾尾表达无显著变化。单侧隐睾手术不影响tPA、uPA和PAI-1 mRNA的表达。结果提示附睾头和附睾体分泌的uPA可能与精子前向运动能力的获得相关。tPA、uPA和PAI-1 mRNA在猕猴附睾头部和体部、前列腺和精囊中的表达可能受睾酮的调节,但不受睾丸分泌因子及温度的影响,且在不同部位睾酮的调节具不同的特征,而附睾尾tPA、uPA和PAI-1的表达则可能是组成性表达。

促性腺激素诱导猕猴排卵周期中卵巢纤溶酶原激活因子与抑制因子的变化

由促性腺激素诱导出的猕猴卵巢tPA(纤蛋白溶酶原激活因子)活性的增加与 排卵密切相关, 排卵前达到高峰, 排卵后明显下降; uPA只在排卵后的颗粒细胞 大量出现; PA的抑制因子PAI-1分泌高峰比tPA峰值早出现12-24h; 排卵来临时, tPA的明显上升导致PAI-1的空然下降。 结果说明: 卵巢中tPA和PAI-1活性的这 种平衡性的变化可能在排卵机制和维持卵巢的正常生理功能中起重要作用, 而uPA 或许与黄体形成的调节有某些关系。图4参17

HORMONAL-REGULATION OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR AND PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-1 IN CULTURED MONKEY SERTOLI CELLS

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5x10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 mu g/cm(2)) in 0.5 mi McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 mi serum-free medium with or without various hormones or other compounds, PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed, Monkey Sertoli cells were also capable of secreting PAI-1, Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity. PAI-1 activity in the culture was also enhanced by these reagents except 8-bromo-dibutyryl-cAMP, forskolin and 3-isobutyl-1-methylxanthin (MIX) which greatly stimulated tPA activity, whereas decreased PAI-1 activity, implying that neutralization of PAI-1 activity by tile high level of tPA in the conditioned media may occur. These data suggest that increased intracellular signals which activate protein kinase A (PKA), or protein kinase C (PKC) can modulate Sertoli cell tPA and PAI-1 activities, The concomitant induction of PA and PAI-1 by the same reagents in the Sertoli cells may reflect a finely tuned regulatory mechanism in which PAI-1 could limit the excession of the proteolysis.

Localization and distribution of tissue type and urokinase type plasminogen activators and their inhibitors type 1 and 2 in human and rhesus monkey fetal membranes

Fetal membranes consist of 10 distinct layers including components of amnion, chorion and decidua, the latter being of maternal origin. They form mechanically integrated sheets capable of retaining amniotic fluid and play an essential role in protecting fetal growth and development in the pregnant uterus. The extracellular matrix, substrate for plasminogen activators (PAs), is an important supportive framework of the fetal membranes. :Fetal membranes from women with preterm premature rupture of membranes may differ in their protease activity compared with normal membranes. To identify the presence of PAs and their inhibitors (PAI) and their possible role in the process of fetal membrane rupture, this study in investigated the distribution and localization of both protein and mRNA for tissue (t) and urokinase (u) PA and their inhibitors type 1 (PAI-1) and type 2 (PAI-2) in amniochorion of human and rhesus monkey using conventional and. confocal immunofluorescence microscopy. In situ hybridization analysis showed that the distribution and localization of mRNAs for tPA, uPA, PAI-I and PAI-2 were similar in the fetal membranes of human and rhesus monkey; no obvious species difference was observed. Evidence of tPA mRNA was detected in amniotic epithelium, trophoblast cells and nearly all cells of the decidual layer. Strong expression of uPA mRNA was noted in the decidual cells which increased in intensity as the abscission point was approached. Weak staining in chorion laeve trophoblast was also detected. In situ hybridization experiments showed PAI-1 mRNA to be concentrated mainly in the decidual cells, some of which were interposed into the maternal-facing edge of the chorion laeve. Maximal labelling of the decidua occurred towards the zone of abscission. Weak expression of PAI-1 mRNA nas also noted in some cells of the chorion laeve. The distribution of PAI-2 mRNA in amniochorion was also concentrated in the cells of the decidual layer, maximum expression of the mRNA was in the level of abscission. No detectable amount of mRNAs for tPA, uPA, PAI-1 and PAI-2 was found in the fibroblast, reticular and spongy layers. Distribution of the proteins of tPA, uPA and PAI-1 in the fetal membranes of these two species was consistent with the distribution of their mRNA. Anti-PAI-2 immunofluorescence was found to be strongly concentrated in the amniotic epithelium, but PAI-2 mRNA was negative in this layer, suggesting that the epithelium-associated PAI-2 is not of epithelial origin. These findings suggest that a local fibrinolysis in fetal membranes generated by precisely balanced expression of PAs and their inhibitors via paracrine or autocrine mechanisms may play an essential role in fetal membrane development, maturation and in membrane rupture. Following an analysis of the distribution and synthesis of activators and inhibitors it was found that they may play a role in abscission during the third stage of labour. (C) 1998 W. B. Saunders Company Ltd.

Synthesis and characterization of poly(amide imide)s containing cyclohexylidene moieties with bulky substituents

Novel poly(amide imide)s (PAI) containing alkyl-substituted cyclohexylidene moieties were synthesized by conventional polycondensation of trimellitic anhydride chloride with novel aromatic diamines followed by chemical imidization using acetic anhydride and pyridine. The inherent viscosities of the resulting PAIs are relatively high and range from 71 to 112 mt g(-1). The prepared PAIs show excellent thermal stability and good solubility. The glass transition temperatures (T-g) measured by DSC are observed in the range of 312-342 degrees C. Furthermore, all the polymers are readily soluble in less hygroscopic organic solvents like cyclohexanone, gamma-butyrolactone as well as aprotic polar solvents.

用稳定性同位素技术确定高寒草甸生态系统中动物营养级模型

根据稳定性同位素技术原理建立了高寒草甸生态系统中动物的营养级模型(3)、(7)和(9)式.3个模型分别描述了每种食物资源对动物的贡献大小(PCV)、食物资源(Ai)占取食动物(P)的整个食物的比例(PAiP)、动物在高寒草甸生态系统中的营养级(TLc):PCVAi=(cos (ΔαPAi))/(ΖPAi)(3)PAiP=(PCVAi)/(∑I=1PCVAi)×%(7)TLc=1+ (αc-αTL1)/Δαcd(9)式中,ΔαPAi为捕食者P与食物Ai的取食角,ΖPAi为捕食向量与食物向量之间的欧氏距离,αc是消费者的向量角,αTL1是第一营级的向量角,利用系数Δαcd是消费者与食物向量角之差(为一常数).同时,给出了判断高寒草甸两个物种之间捕食或营养关系模型(Ζ_(S_1S_2)):当cos(Δα)/PCVmin≤Ζ_(S_1S_2)≤cos(Δα)/PCVmax时存在捕食关系,并为上下级营养关系;当Ζ_(S_1S_2)cos(Δα)/PCVmax时,不存在捕食关系,前式为同一营养级,后式为相隔一个至几个营养级.模型(9)式得到的结果与张晓爱等(1999)报道一致.

东喜马拉雅构造结新生代碰撞变形与岩石圈演化

The Eastern Himalayan Syntaxis (EHS) is one of the strongest deformation area along the Himalayan belt resulted from the collision between Indian plate and the Eurasian Plate since the 50～60Ma, and has sensitivity tracked and preserved the whole collisional processes. It should depend on the detail geological investigations to establish the deformational accommodate mode, and the uplift history, to elucidate the deep structure and the crust-mantle interaction of the Tibet Plateau of the EHS. The deep-seated (Main Mantle Thrusts) structures were exhumed in the EHS. The MMT juxtapose the Gangdese metamorphic basement and some relic of Gangdese mantle on the high Himalayan crystalline series. The Namjagbawa group which is 1200～1500Ma dated by U/Pb age of zircon and the Namla group which is 550Ma dated by U/Pb age of zircon is belong to High Himalayan crystalline series and Gangdese basement respectively. There is some ophiolitic relic along the MMT, such as metamorphic ocean mantle peridotite and metamorphic tholeiite of the upper part of ocean-crust. The metamorphic ocean mantle peridotites (spinel-orthopyroxene peridotite) show U type REE patterns. The ~(87)Sr/~(86)Sr ratios were, 0.709314～0.720788, and the ~(143)Nd/~(144)Nd ratios were 0.512073～0.512395, plotting in the forth quadrant on the ~(87)Sr/~(86)Sr-~(143)Nd/~(144)Nd isotope diagram. Some metamorphic basalt (garnet amphibolite) enclosures have been found in the HP garnet-kynite granulite. The garnet amphibolites can be divided two groups, the first group is deplete of LREE, and the second group is flat or rich LREE, and their ~(87)Sr/~(86)Sr, ~(143)Nd/~(144)Nd ratios were 0.70563～0.705381 and 0.512468～0.51263 respectively. Trace element and isotopic characteristics of the garnet amphibolites display that they formed in the E-MORB environment. Some phlogolite amphibole harzburgites, which exhibit extensive replacement by Phl, Amp, Tc and Dol etc, were exhumed along the MMT. The Phl-Amp harzburgites are rich in LREE and LILE, such as Rb, K etc, and depletes Eu (Eu~* = 0.36 ～ 0.68) and HFSE, such as Nb, Ta, Zr, Hf, P, Ti etc. The trace element indicate that the Phl-Amp harzburgites have island arc signature. Their ~(87)Sr/~(86)Sr are varied from 0.708912 to 0.879839, ~(143)Nd/~(144)Nd from 0.511993 to 0.512164, ε Nd from- 9.2 to - 12.6. Rb/Sr isochrone age of the phlogolite amphibole harzburgite shows the metasomatism took place at 41Ma, and the Amp ~(40)Ar/~(39)Ar cooling age indcate the Phl-Amp harzburgite raising at 16Ma. There is an intense crust shortening resulted from the thrust faults and folds in the Cayu block which is shortened more 120km than that of the Lasha block in 35～90Ma. With the NE corner of the India plate squash into the Gangdese arc, the sinistral Pai shear fault and the dextral Aniqiao shear fault on the both sides of the Great bent of Yalun Zangbu river come into active in 21～26Ma. On the other hand, the right-lateral Gongrigabu strike-slip faults come into activity at the same period, a lower age bound for the Gongrigabu strike-slip fault is estimated to be 23～24Ma from zircon of ion-probe U/Pb thermochronology. The Gongrigabu strike-slip faults connect with the Lhari strike-slip fault in the northwestern direction and with the Saganing strike-slip at the southeastern direction. Another important structure in the EHS is the Gangdese detachment fault system (GDS) which occurs between the sedimental cover and the metamorphic basement. The lower age of the GDS is to be 16Ma from the preliminary 40Ar/39Ar thermochronology of white mica. The GDS is thought to be related to the reverse of the subducted Indian crust and the fast uplift of the EHS. Structural and thermochronology investigation of the EHS suggest that the eastern Tibet and the western Yunnan rotated clockwise around the EHS in the period of 35～60Ma. Later, the large-scale strike-slip faults (RRD, Gaoligong and Saganing fault) prolongate into the EHS, and connect with the Guyu fault and Gongrigabu fault, which suggest that the Indianchia block escape along these faults. Two kind of magmatic rocks in the EHS have been investigated, one is the mantle-derived amphibole gabbro, dioposide diorite and amphibole diorite, another is crust origin biotit-garnet adamellite, biotit-garnet granodiorite and garnet-amphibole-biotite granite. The amphibole gabbro dioposite diorite and amphibole diorite are rich in LREE, and LILE, such as Ba, Rb, Th, K, Sr etc, depleted in HFSE, such as Nb, Ta, Zr, Hf, Ti etc. The ratio of ~(87)Sr/~(86)Sr are from 0.7044 to 0.7048, ~(143)Nd/~(144)Nd are from 0.5126 to 0.5127. The age of the mantle origin magamatic rocks, which result from the partial melt of the raising and decompression anthenosphere, is 8Ma by ~(40)Ar/~(39)Ar dating of amphibole from the diorite. The later crust origin biotite-garnet adamellite, biotite-garnet granodiorite and garnet-amphibole-biotite granite are characterized by aboudance in LREE, and strong depletion of Eu. The ratios of ~(87)Sr-~(86)Sr are from 0.795035 to 0.812028, ~(143)Nd/~(144)Nd from 0.51187 to 0.511901. The ~(40)Ar/~(39)Ar plateau age of the amphibole from the garnet-amphibole-biotite granite is 17.5±0.3Ma, and the isochrone age is 16.8±0.6Ma. Their geochemical characteristics show that the crust-derived magmatic rocks formed from partial melting of the lower curst in the post-collisional environment. A group of high-pressure kaynite-garnet granulites and enclave of high-pressure garnet-clinopyroxene grnulites and calc-silicate grnulites are outcroped along the MMT. The peak metamorphic condition of the high-pressure granulites yields T=800～960 ℃, P=1.4～1.8Gpa, corresponding the condition of 60km depth. The retrograde assemblages of the high-pressure grnulites occur at the condition of T=772.3～803.3 ℃, P=0.63～0.64Gpa. The age of the peak metamorphic assemblages are 45 ～ 69Ma indicated by the zircon U/Pb ion-plobe thermochronology, and the retrograde assemblage ages are 13～26Ma by U/Pb, ~(40)Ar/~(39)Ar thermochronology. The ITD paths of the high-pressure granulites show that they were generated during the tectonic thickening and more rapid tectonic exhumation caused by the subducting of the Indian plate and subsequent break-off of the subducted slab. A great deal of apatite, zircon and sphene fission-track ages, isotopic thermochronology of the rocks in the EHS show that its rapid raising processes of the EHS can be divided into three main periods. There are 35～60Ma, 13～25Ma, 0～3Ma. 3Ma is a turn in the course of raising in the EHS which is characterized by abruptly acceleration of uplifting. The uplift ratios are lower than 1mm .a~(-1) before 3Ma, and higher than 1mm .a~(-1) with a maximum ratio of 30mm .a~(-1) since 3Ma. The bottom (knick point) of the partial anneal belt is 3.8km above sea level in the EHS, and correspond to age of 3Ma determined by fission-track age of apatite. The average uplift ratio is about 1.4 mm .a~(-1) below the knick point. The EHS has raised 4.3km from the surface of 2.36km above sea level since 3Ma estimated by the fossil partial anneal belt of the EHS. We propose a two-stage subduction model (B+A model) basing on Structural, thermochronological, magmatical, metamorphic and geophysical investigations of the EHS. The first stage is the subduction of the Indian continental margin following after the subduction of the Tethys Ocean crust and subsequent collision with the Gangdese arc, and the second stage is the Indian crust injecting into the lower crust and upper mantle of the Tibet plateau. Slab break-off seems to be occurred between these two stages.