新疆药用延胡索资源调查

作者对新疆维吾尔自治区的9个市、县进行了药用延胡索的资源调查,采集植物标本70余份,弄清药用种类主要是3种,即Corydalis glaucescens Regel,C.ledebouriana Kir.et Kar.和C.schanginii(Pall.)Fedtsch.;初步了解了3种药用延胡索植物的分布状况,并对块茎中总生物碱的含量作了分析。     

Thrombin increases proliferation and decreases fibrinolytic activity of kidney glomerular epithelial cells.

Human glomerular epithelial cells (GECs) in culture synthesize single-chain, urokinase-type plasminogen activator (SC-uPA), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor 1 (PAI-1) and possess specific membrane-binding sites for u-PA. Using purified 125I-alpha thrombin, we demonstrate here the presence of two populations of specific binding sites for thrombin on GECs (1.Kd = 4.3 +/- 1.0 x 10(-10) M, 5.4 +/- 1.4 x 10(4) M sites per cell, 2. Kd = 1.6 +/- 0.5 x 10(-8) M, 7.9 +/- 1.8 x 10(5) sites per cell). Purified human alpha thrombin promoted the proliferation of GECs and induced a time- and dose-dependent increase of SC-uPA, t-PA, and PAI-1 antigens released by GECs. Thrombin-mediated increase in antigen was paralleled by an increase in the levels of corresponding u-PA and PAI-1 messenger RNA. In contrast, thrombin decreased u-PA activity in conditioned medium. This discrepancy between u-PA antigen and u-PA activity was explained by a limited proteolysis of SC-uPA by thrombin, leading to a two-chain form detected by immunoblotting and that could not be activated by plasmin. Thrombin also decreased the number of u-PA binding sites on GECs (p less than 0.05) without changing receptor affinity. Hirudin inhibited the binding and the cellular effects of thrombin, whereas thrombin inactivated by diisopropylfluorophosphate had no effect, indicating that both membrane binding and catalytic activity of thrombin were required. We conclude that thrombin, through specific membrane receptors, stimulates proliferation of GECs and decreases the fibrinolytic activity of GECs both at the cell surface and in the conditioned medium. These results suggest that thrombin could be involved in the pathogenesis of extracapillary proliferation and persistency of fibrin deposits in crescentic glomerulonephritis.     

Micromanipulation of mitotic chromosomes in PTK2 cells using laser-induced optical forces ("optical tweezers")

To study the potential use of optical forces to manipulate chromosome movement, we have used a Nd:YAG laser at a wavelength of 1.06 microns focused into a phase contrast microscope. Metaphase and anaphase chromosomes were exposed while being monitored by video microscopy. The results indicated that when optical forces were applied to late-moving metaphase chromosomes on the side closest to the nearest spindle pole, the trapped chromosomes initiated movement to the metaphase plate. The chromosome velocities were two to eight times the normal rate depending on the chromosome size, geometry, and trapping site. At the initiation of anaphase, a pair of chromatids could be held by the optical trap and kept motionless throughout anaphase while the other pairs of chromatids separated and moved to opposite spindle poles. As a result, the trapped chromosome either was incorporated into one of the daughter cells or was lost in the cleavage furrow, or the two chromatids eventually separated and moved to their respective daughter cells. If the trap was removed at the beginning of anaphase B, the chromosome moved back to the poles. Our experiments demonstrate that the laser-induced optical force trap is a potential new technique to study noninvasively the mitotic spindle of living cells.     

禿杉组织培养研究

本文详细报道了从秃杉(Taiwania flousiana Gaussen)离体胚诱导不定芽、不定根及从无菌苗茎端培养再生植株的过程。诱导不定芽要求较低的蔗糖浓度(以3％最好);同时BA是必须的,在附加0.1—3 mg/1 BA的White培养基上,从离体胚的子叶或胚轴上诱导了不定芽的发生(以1 mg/1最好);NAA与BA结合使用,对不定芽诱导无促进作用;适当提高光照有利于不定芽的诱导。在诱导不定芽的同时,在子叶表面还观察到有许多无结构的“不定突起”。不定芽起源于子叶表皮下1—2层细胞。IBA对诱导离体胚上产生不定根效果较好。在有或无生长素的培养基上,从生长1月龄的无菌苗茎端培养获得了不定根的产生,在加有细胞分裂索的培养基上,从无菌苗上产生了腋芽。     

层粘连蛋白与顺铂对腹水肝癌细胞内微丝的共同作用关系

本文探讨在外源性层粘连蛋白与抗癌药物顺铂的共同作用下,癌细胞内微丝组装的变化。结果发现外源性层粘连蛋白与小鼠腹水型肝癌细胞膜受体结合后,促进肌动蛋白微丝组装,使其含量增加;而多靶性抗癌药物顺铂与肌动蛋白微丝的结合,抑制微丝组装过程,造成微丝含量减少;两种试剂共同作用于癌细胞时,肌动蛋白微丝的含量与对照组相比非常接近。本研究为上述两种物质对癌细胞内微丝组装的拮抗性作用提出直接证据。     

3-磷酸甘油醛脱氢酶胍变性时的活力及构象变化

酵母3-磷酸甘油醛脱氢酶在盐酸胍溶液中的内源荧光及剩余活力的变化结果提示:apo酶及holo酶的活力在胍浓度为0.5M左右可完全丧失.同时伴有内源荧光强度的下降,光谱宽度的增加和335nm最大发射峰的红移(提示了色氨酸残基的暴露).与已经报导的肌肉酶(内源荧光强度在胍浓度为0.4—1.2M范围相对稳定)不同,酵母酶内源荧光在此浓度范围内表现为逐渐降低.在0.7M胍溶液中,内源荧光变化动力学过程只能测出一相,而酶失活动力学过程为快慢两相,快相动力学速度常数至少大于内源荧光降低速度常数三个数量级以上.以上结果提示:低浓度胍可引起该酶的完全失活,活性部位的空间构象比酶分子的构象更易受到变性剂的扰乱;有一个色氨酸残基位于或靠近酶的活性部位.     

急进高海拔时健康人夜间睡眠、呼吸状态和血氧饱和度的变化

在海拔2300m选择健康成年男性5人,急进抵海拔4660m,用多导监测仪分别在两地连续7h监测夜间睡眠、呼吸状态和血氧饱和度变化,进行自身对比。结果发现:(1)急进高海拔后,总睡眠时间、有效睡眠指数、Ⅲ～Ⅳ期深睡眠均较中度高原减少(p<0.01);总觉醒时间、Ⅰ～Ⅱ期浅睡眠高海拔较中度高原增多(p<0.05):(2)急进高海拔后,有3名健康人出现周期性呼吸,其中1名健康者出现周期性呼吸119次,伴有中枢性睡眠呼吸暂停,最低Sao_2为78％;(3)同海拔高度夜间睡眠时与清醒时Sao_2相比较,中度高原下降4.2％,高海拔下降11.2％(p<0.01);高海拔与中度高原夜间清醒时Sao_2相比较下降7.4％,睡眠时下降14.4％(p<0.001)。结果提示:(1)睡眠加重了高原人原有的低氧血症;(2)低氧血症导致睡眠结构的紊乱和睡眠质量的降低;(3)睡眠中出现的周期性呼吸,应视为机体的一种自我保护机制;(4)频发的周期性呼吸或睡眠呼吸暂停将影响大脑机能。     

多肽的α螺旋结构对多肽与钙调蛋白亲合力的影响

本文设计并采用固相法合成了4种钙调蛋白可结合多肽,这些多肽分成两组,每一组中两个多肽的碱性和疏水性相近,但形成α螺旋结构的倾向(预测)不同。研究了这些多肽与钙调蛋白的相互作用,在Ca~(2+)存在下,这些多肽与丹磺酰钙调蛋白结合,使丹磺酰钙调蛋白的荧光光谱发生显著变化,测定了多肽与钙调蛋白所形成的复合物的解离常数。结果表明,预测形成α螺旋结构倾向较大的多肽与钙调蛋白的亲合力也较大。     

Bam HI DNA甲基化酶酶学性质的研究

以Lineweave-Burk plot双倒数作图法测得该酶对底物S-腺苷酰甲硫氨酸(SAM)的K_m=7.69×10~(-6)mol/L,在1mmol/LS-腺苷酰高半胱氨酸(SAH)存在下,Ki=7.33×10~(-4)mol/L,两条直线相交于纵轴,证明SAH是该酶的竞争性抑制剂。该酶最适pH为7.8,对热不稳定。同时还测定了该酶对不同DNA底物的专一性及盐浓度、代谢相关物’两价阳离子、某些酸根等对该酶调节性质的影响。以碘代乙酰胺修饰该酶的SH基’及用二硫苏糖醇(DTT)和巯基乙醇(MSH)保护该酶SH基所作的实验表明SH基是该酶活性中心所必需的,用高效液相色谱(HPLC)法证明该酶所甲基化的碱基为刘氏小球菌(M·L、DNA)分子中的胞嘧啶,且求得甲基化30min后所得甲基化水平为2.39％。同时也证明当用该酶将λDNA甲基化后,可使BamHI限制性核酸内切酶对甲基化后的λDNA丧失切割作用。