羊栖菜褐藻糖胶抗凝血活性的研究

本文研究了羊栖菜褐藻糖胶的化学组成和抗凝血活性之间的关系。采用热水提取得羊栖菜粗多糖,CaCl2纯化得褐藻糖胶,DEAE Sepharose CL-6B柱层析与Sepharose CL-6B柱层析对褐藻糖胶进行分级,得到F1、F2、F31、F32和F33五个级分,均为岩藻糖、半乳糖和甘露糖等糖基组成的杂多糖,并含有硫酸酯和糖醛酸以及少量的蛋白质,相对分子质量范围2．5万～95万。采用活化部分凝血活酶时间(APTT)和凝血酶时间(TT)检测了这5个级分的抗凝血活性,结果显示,羊栖菜褐藻糖胶能显著延长APTT的凝血时间,而对TT的影响不明显。F1、F31和F32对APTT的影响比较显著,而F2、F33和羊栖菜粗多糖的影响较小。研究表明,羊栖菜褐藻糖胶主要是通过抑制内源凝血途径而达到抗凝血的效果,其抗凝血活性与褐藻糖胶的硫酸基含量成正相关,而与相对分子质量和糖醛酸含量无关。     

裙带菜褐藻糖胶的分离纯化及其结构分析

裙带菜经水提法提取得到褐藻糖胶粗多糖,经DEAE-Sepharose FF离子交换层析和 Sepharose 4B层析后,得到Sl、S2两个单一组分,相对分子质量分别为550 808、38 335.基本结构及单糖组成分析表明,二者均含有岩藻糖、糖醛酸、硫酸基、半乳糖、甘露糖、葡萄糖、鼠李糖、木糖、阿拉伯糖,但含量差别较大,推测与二者的生理活性的差异有关.     

林肯链霉菌丙氨酸脱氢酶的纯化和性质

采用硫酸铵分级沉淀、DEAE-纤维素52柱层析、亲和蓝柱层析和琼脂糖凝胶Sepharose6B柱层析的方法,分离纯化了林肯链霉菌丙氨酸脱氢酶,用聚丙烯酰胺凝胶电泳鉴定为单一组分。以凝胶过滤和聚丙烯酰胺梯度凝胶电泳测得该酶的分子量为170000,SDS-聚丙烯酰胺凝胶电泳测得其亚基分子量为42500,表明林肯链霉菌丙氨酸脱氢酶由四个相同的亚基组成。该酶加氨反应最适pH为9．0,脱氨反应最适pH为9．5,加氨反应和脱氨反应的最适温度均为50℃。加氨反应丙氨酸脱氢酶的表现米氏常数km值为：丙酮酸2．08×10－4mol／L,NH4+2．00×10-2mol／L,NADH2．38×10-5mol／L;脱氨反应的Km为：L-Ala1．43×10-2mol／L;NAD+6．67×10-5mol／L。     

香菇多糖提取分离的研究

采用DEAE—cellulose柱层析和Sephadex G-200柱层析从香菇子实体热水浸提的粗多糖中分离出两个多糖组分Lenl—A、Len2-A,经旋光度法、Sepharose 4B柱层析、红外光谱等方法检测确定为均一多糖组分,且均为α-构型的吡喃糖,苯酚硫酸法测得其糖质量分数分别为91．5％和92．3％,旋光度为＋5．6°、＋11．2°,相对分子质量为375×10^3、718×10^3。     

甜菜坏死黄脉病毒的分离提纯及其生物化学特性的研究

本研究工作中,建立了一个有效的甜菜坏死黄脉病毒的分离提纯程序,解决了该病毒粒体易于聚集难以提纯的问题,其操作要点是,(1)通过Sepharose 2B柱层析代替超离心,有效地除去一些小分子量核酸杂质;(2)经PEG再次沉淀浓缩后,调整pH至酸牲(pH3.0),使病毒充分悬浮以减少凝聚;(3)在病毒等电点(pH4.8～4.9)条件下,进一步沉淀以纯化病毒。根据病毒提取物的OD260/OD280比值,算出核酸含量约4.5％。核酸电泳出现4条带,分子量分别为:2.25×10~(?),1.8×10~(?),1.05×10~(?),0.75×10~(?)道尔顿。病毒提取物经超速离心出现4个界面,沉淀系数分别为,200.8S,165S,125.8S,100S。说明甜菜坏死黄脉病毒可能是4组分病毒粒体。病毒粒体含一蛋白亚基,分子量约为2.05±0.05×10~4道尔顿,由16种共199个氨基酸组成。     

从黑曲霉提取甲壳素和壳聚糖

采用电解法从培养的黑曲霉湿菌体制甲壳素 ;采用碱提取法从培养的黑曲霉湿菌体制壳聚糖。甲壳素提取的得率干菌重量的 2 0 6 %。所得干燥壳聚糖产品的游离胺基为 93 76 % ,0 5%壳聚糖的 0 5%醋酸的运动粘度为 5 4 4 8× 10 6 m2 /s ,粘均分子量为 8 2 75× 10 4 ,含水量为 9 16 % ,产品得率为 12 11%。     

两种有鳞类爬行动物消化道微生物的研究

从北草蜥的胃、小肠和大肠中分离出 7个属 8种细菌和 2个属的霉菌 ,从中国石龙子的胃、小肠和大肠中分离到 6个属 7种细菌和 3个属的霉菌 ,酵母菌和放线菌都未发现。北草蜥胃、小肠和大肠中的细菌数量分别为 2 2 7× 1 0 6,2 0 6× 1 0 6和 9 78× 1 0 8个 / g ;霉菌数量为 2 5× 1 0 4 ,2 1 5× 1 0 4 ,1 33× 1 0 4个 /g。细菌在中国石龙子胃、小肠和大肠中的数量分布依次为 3 51× 1 0 6,2 2 5× 1 0 6,1 4 2× 1 0 9个 /g;霉菌分布数量为 2 1 9× 1 0 4 ,1 67× 1 0 4 和 1 61× 1 0 4 个 / g     

阿魏侧耳多糖的分离纯化与抗肿瘤活性的研究*

从阿魏侧耳 (Pleurotusferulae)子实体中用热水浸提出粗多糖PFW ,将PFW脱蛋白后 ,经DEAE—纤维素柱层析 ,得两种多糖PF1 和PF2 。用光散射和GPC联机测定 :PF1 含有两种组分其重均分子量分别为 7 875× 1 0 5 和 3 2 4 5× 1 0 4 ;PF2 重均分子量为 3 1 87× 1 0 6。用IR和GC分析它们组成和结构 ,结果表明 :PF1 由鼠李糖、葡萄糖、半乳糖和甘露糖组成 ,含有 β 糖苷键和甘露糖苷 ;PF2 由鼠李糖、木糖、葡萄糖、半乳糖和甘露糖组成 ,含有α 糖苷键。给小鼠腹腔注射多糖 ,对移植性肿瘤S 1 80均有一定的抑制作用     

HPLC法测定麻仁润肠丸大黄素、大黄酚的含量

采用KromacilC18(4 .6mm× 2 5 0mm ,5 μm)色谱柱 ,以甲醇 - 0 .1%磷酸溶液 (85∶15 )为流动相 ,流速为 1mL·min ,柱温为 30℃ ,检测波长为 2 5 4nm ,以外标法测定了麻仁润肠丸中大黄素、大黄酚的含量。大黄素在 8.992× 10 -3 ～ 116 .896× 10 -3 ,大黄酚在 2 1.376× 10 -3 ～ 2 77.85 8× 10 -3 范围内呈线性关系 ,其在制剂中的平均回收率 (n =6 )分别为 10 1.5 6 % (RSD =1.3% )、96 .78% (RSD =1.3% )。     

发状念珠藻胞外多糖的纯化与性质分析

采用DEAE阴离子交换层析和Sephadex G100凝胶层析对液体悬浮培养发状念珠藻胞外多糖进行纯化, 得到两个组分NFPS1和NFPS2。对组分NFPS2进行理化性质分析, 并与野生发状念珠藻多糖NFPS0的性质进行对比。结果表明二者具有相似的单糖组成, 均为葡萄糖、木糖、半乳糖、甘露糖; 表观分子量分别为2.79×105、2.26×105; 均不含核酸、蛋白质等物质, 是非硫酸化多糖; 有较高的热稳定性, 其降解温度在245oC左右。但在微观结构上, 两者存在一定差别。     

Characteristics of the membranes of the pellicle of the ciliatePseudomicrothorax dubius

Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.     

The early development of the tail and the transformation of the shape of the nucleus of the spermatid of the domestic fowl,Gallus gallus

Summary The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl.In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum.The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.The authors wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices     

Characterization of the composition of the aqueous humor and the vitreous body of the eye of the frog Rana temporaria L

This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at ~ 1.5 × 10^− 4 mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed.     

Observations on the permeability of the choriocapillaris of the eye

Summary The choriocapillaris is a fenestrated capillary bed located posterior to the retinal pigment epithelium. It serves as the main source of supply to the photoreceptors, retinal pigment epithelium, and other cells of the outer retina. The permeability of these capillaries to intravenously injected ferritin (MW — approx. 480,000; mol. diam. 11 nm) was examined in the mouse, rabbit, and guinea pig, each of which is characterized by a different type of retinal vascularization. In all three species, the bulk of the ferritin remained in the capillary lumina, where it appeared to be blocked at the level of the diaphragmed fenestrae. Some ferritin was present in endothelial cell vacuoles. The results confirm previous work on the rat choriocapillaris and indicate that the barrier function of the choriocapillary endothelium is present even among species in which the retinal circulation differs significantly.Supported by NIH grant EY03418