陕北安塞退耕地持久土壤种子库与地上植被的对应关系

通过研究陕北安塞退耕3～31 a 的阴阳坡10 个样地的持久土壤种子库特征及其与地上植被的关系,分析持 久土壤种子库在植被恢复中的作用。结果表明:调查样地持久土壤种子库由15 个科36 个物种组成,种子密度在 1 908～12 617粒/ m2 之间,1 年生物种占80 % ,随着退耕演替的进行,种子库中1 年生物种比例呈下降趋势,多年生 物种比例呈上升趋势,地带性物种在持久种子库中的比例也随退耕时间的延长而增加,物种数和种子库密度均随 退耕演替先增大后降低;持久土壤种子库与地上植被的相似性较低,Serensen 相似性系数为0121～0142 ;通过对适 宜物种的确限度及土壤种子库大小的分析,认为该区退耕地的持久土壤种子库具有一定的植被恢复潜力。

长期施用氮、磷和有机肥对不同种植体系土壤有效硫累积的影响

以22年定位试验为基础,研究了长期施用氮、磷和有机肥(NPM)对不同种植体系土壤有效硫在剖面上分布与累积状况。结果表明,在60—80 cm土层各处理出现第一个累积峰,累积峰值为粮饲轮作31.3,玉米连作29.2,小麦连作27.9,粮豆轮作25.6,苜蓿连作24.0 mg/kg;在140—180 cm土层各处理又出现有效硫的第二个累积峰,累积峰值为粮饲轮作44.7,粮豆轮作43.1,小麦连作41.0,玉米连作39.7,苜蓿连作36.5 mg/kg。第二累积峰值均大于第一累积峰值。0—200 cm土层有效硫总累积量粮饲轮作高达746.3 kg/hm~2,其次为玉米连作640.6,粮豆轮作为638.3,小麦连作为622.4,苜蓿连作最小为557.3 kg/hm~2。长期施用磷肥和有机肥是有效硫在土壤中累积的主要因素,有效硫在土壤剖面上有向深层迁移的趋势。不同作物对硫的吸收利用差异和不同种植方式对有效硫的累积与分布产生影响。

WEPP模型坡面版在黄土丘陵沟壑区的适用性评价——以坡度因子为例

利用安塞试验站1985-1992年的气象观测数据和野外坡度径流小区径流和土壤侵蚀量监测资料,评价了WEPP模型在黄土丘陵沟壑区不同坡度条件下的适用性。结果表明,径流量的模拟值在10°、15°、20°、25°和28°五个坡度条件下,变化幅度不如实测值明显,但模拟值随坡度变化的趋势和实测值相一致;WEPP模型对次降雨、年平均和多年平均土壤侵蚀量模拟结果较好,且无论是模拟值在不同坡度之间的差值、还是模拟值随坡度变化趋势均与实测值接近。I30对WEPP模型模拟次降雨径流量有重要影响,当I30大于0.92mm/min,模型模拟误差较大。WEPP模型对次降雨土壤侵蚀量的模拟与PI30密切相关,当PI30大于129mm2/min时,模型模拟误差较大。∑PI30对WEPP模型模拟年平均径流量和侵蚀量有重要影响,当∑PI30大于150mm2/min时,模拟精度明显下降,且∑PI30对径流模拟影响较对土壤侵蚀模拟影响明显。WEPP模型对次降雨、年平均和多年平均径流量模拟的Nash-Sutcliffe有效性ME分别为0.914、0.912和0.617;对次降雨、年平均和多年平均土壤侵蚀量模拟的Nash-Sutcliffe有效性ME...

高效毛细管电泳安培检测的应用研究

毛细管电泳在生命科学，生物技术，医学药物和环境保护中显示了极其重要的应用前景，也被认为是人类进入纳米技术时代的一种富有重要潜在价值的手段。具有高效、快速，耗样量小等特点。而安培电化学恰恰适合毛细管电泳对检测器的要求。毛细管电泳-安培检测法在多方面发挥着不可忽视的作用。本文用此方法进行了一些分析。1. 毛细管区带电泳柱端安培检测抗癌药物巯嘌呤 报道了一种高效快速检测抗癌药物巯嘌呤的方法-毛细管区带电泳柱端安培检测。优化了检测条件，考察了巯嘌呤的电化学行为。在15kV电压下进样3s，在此电压下进行电泳分离，背景电解质为10mmol/L Na_2HPO_4，1.2V检测时，此药物的检测限为1 * 10~(-7)mol/L，线性范围为5 *　10~(-4)-5　* 10~(-6)mol/L，相关系数为0.995，重现性良好。并用该法检测了人体尿样及牛血清蛋白中的巯嘌呤，本工作具有一定的临床意义。2. 毛备管区带电泳柱端安培分离分析多种抗癌药物 通过毛细管区带电泳柱端安培检测法对8-氮鸟嘌呤，5-氟尿嘧啶，6-巯嘌呤，氨甲喋呤和6-硫鸟嘌呤五种抗癌药物进行了灵敏，高效，迅速的分离检测。实验采用33μm的碳纤维盘电极，在20min内分离检测了此五种药物的混合物。在1.2V的检测电位下，五种药物的线性范围超过了三个数量级，相关系数是0.992，这五种药物的检测限依次为5，3，5，9和5 * 10~(-7)mol/L。3. 三种噻唑衍生物的毛细管电泳安培分析 在碳纤维电极上检测三种噻唑衍生物，2-氨基噻唑，2-氨基苯并噻唑和2-巯基苯并噻唑。对应检测限分别为2.0，0.013，0.025 * 10~(-6)mol/L。它们的线性范围较宽，相关系数均在0.9995以上。分别考察了缓冲液pH值及其浓度，分离电压对电泳分离效果的影响，还对实际样品进行了分析。此工作具有一定的实用意义。

工艺流程中高浓定影液酸度的流线分析方法研究

每种酸碱指示剂在其变色PH范围内都有一个明显的颜色变化，在其相应的PH-A曲线上总存在一段直线部分，通过选择一定PH变色范围和适当浓度的指示剂，就能使各指示剂PH-A曲线上的直线部分相互迭加成一条新的直线，从而加宽了混合指示剂的表现变色PH范围，依此原理，本论文以二种流动注射分析方法独立地完成了工艺流程中高浓定影液酸度的在线分析。并对这二种方法做了简单的对比。（一）工艺流程中高浓定影液酸度的在线分析方法研究（1）我们选择了内含0.08%溴甲酚紫，0.04%溴百里酚兰，0.02%甲酚红组成的混合指示剂的6%稀释液作为试剂，以C-41CX定影浓缩液作载流，用F2A-分光光度法完成了高浓度定影液酸度的在线测定，相对标准偏差在50.2%以内，采样频率为100次/h，分析周期很短（25秒），在5.5～8.0PH范围内，PH-A工作曲线具有很好的线性关系，相关系数0.9996，斜率即灵敏度达到：0.198A/PH。（二）工艺流程中高浓定影液酸遮掩在线分析方法研究（2）我们造反了0.005%的溴甲酚紫，0.009%的溴百里酚兰和0.025%的甲酚红作为混合酸碱指示剂，以蒸馏水作载流，通过选择最佳测定条件用流动注射-分光光度法（F2A-Spectrophotometry）在线测定了不宜用其它分析法测定的高浓溶液的酸度。该分析方法还可以推广到其它高浓溶液的酸度的在线测定。相对误差在±0.2%以内；分析周期为30秒；采样体积为60μL。在5.2～8.5PH范围内，混合指示剂的PH-A工作曲线具有良好的线性关系，相关系数达到0.9991，工作曲线斜率即是灵敏度达到0.376A/PH。

GaN基垂直腔面发射激光器的研制

作者在我国大陆首次实现了GaN基垂直腔面发射激光器（VCSEL）的激射．首先采用金属有机物化学气相沉积（MOCVD）技术在蓝宝石衬底上进行高质量氮化物增益区的外延生长．然后在表面沉积高反射介质膜分布布拉格反射镜（DBR）．将样品键合到其它支撑片上后，采用激光剥离技术将蓝宝石衬底去除．再在去除蓝宝石后露出的氮化物表面沉积第二组介质膜DBR制成VCSEL．在室温光泵条件下，观察到VCSEL的激射．激射波长449．5nm。阈值6．5mJ／cm~2．激射峰的半高宽小于0．1nm，这些指标达到了国际领先水平．本文为进一步研制实用化氮化物VCSEL奠定了重要的基础．

HCl/HF/CrO3溶液对 InGaAs/InGaAsP的选择性湿法刻蚀--应用于楔形结构的制备

利用动态掩膜湿法腐蚀技术,研究了HCl/HF/CrO3溶液对与InP衬底晶格匹配的InxGa1-xAs1-yPy(y=0,0.2,0.4,0.6)材料的腐蚀特性.对于HCl(36wt%)/HF(40wt%)/CrO3(10wt%)的体积比为x∶0.5∶1的溶液,随着x由0增加到1.25,相应的腐蚀液对In0.53Ga0.47As/In0.72Ga0 28As06P0.4的选择性由42.4降到1.4;通过调节腐蚀液的选择性,在In072Ga0.28As06P0.4外延层上制备出了倾角从1.35°到35.9°的各种楔形结构;当x为0.025和1.25时,相应的In0.72Ga0.28As0.6P0.4腐蚀表面的均方根粗糙度分别为1.1nm和1.6nm.还研究了溶液的组分与InxGa1-xAs1-yPy(y=0,0.2,0.4)的腐蚀速率间的关系,并对腐蚀机理进行了分析.

Mechanism Responsible for the Complete Suppression of Karman Vortex in Flows Past a Wavy Square-Section Cylinder

The Karman vortex shedding is totally suppressed in flows past a wavy square-section cylinder at a Reynolds number of 100 and the wave steepness of 0.025. Such a phenomenon is illuminated by the numerical simulations. In the present study, the mechanism responsible for it is mainly attributed to the vertical vorticity. The geometric disturbance on the rear surface leads to the appearance of spanwise flow near the base. The specific vertical vorticity is generated on the rear surface and convecting into the near wake. The wake flow is recirculated with the appearance of the pair of recirculating cells. The interaction between the upper and lower shear layers is weakened by such cells, so that the vortex rolls could not be formed and the near wake flow becomes stable.

Intercellular calcium wave propagation in linear and circuit-like bone cell networks

In the present study, the mechanism of intercellular calcium wave propagation in bone cell networks was identified. By using micro-contact printing and self-assembled monolayer technologies, two types of in vitro bone cell networks were constructed: open-ended linear chains and looped hexagonal networks with precisely controlled intercellular distances. Intracellular calcium responses of the cells were recorded and analysed when a single cell in the network was mechanically stimulated by nano-indentation. The looped cell network was shown to be more efficient than the linear pattern in transferring calcium signals from cell to cell. This phenomenon was further examined by pathway-inhibition studies. Intercellular calcium wave propagation was significantly impeded when extracellular adenosine triphosphate (ATP) in the medium was hydrolysed. Chemical uncoupling of gap junctions, however, did not significantly decrease the transferred distance of the calcium wave in the cell networks. Thus, it is extracellular ATP diffusion, rather than molecular transport through gap junctions, that dominantly mediates the transmission of mechanically elicited intercellular calcium waves in bone cells. The inhibition studies also demonstrated that the mechanical stimulation-induced calcium responses required extracellular calcium influx, whereas the ATP-elicited calcium wave relied on calcium release from the calcium store of the endoplasmic reticulum.

四川九顶山西坡及龙肘山维管植物区系和植物多样性研究

横断山地区是一个十分自然的植物区系地区，在中国植物区系分区中是作为泛北极植物区中国-喜马拉雅亚区中的一个地区，其种子植物区系具有丰富的科、属、种，地理成分复杂，特有现象和替代现象明显。该地区作为植物区系和生物多样性的研究热点地区，长期以来极受中外植物学家关注。横断山脉东缘是中国-喜马拉雅和中国-日本植物区系的交汇过渡区域，北部的岷江流域以及南部的金沙江流域，孕育了该区丰富的物种资源和植被资源。而岷江干热河谷和金沙江干热河谷的相似性和相关性，更为该区的植物区系和生物多样性南北的对比研究提供了有利的条件。 本研究选择的九顶山西坡和龙肘山分别位于横断山区北部和南部，九顶山属岷江流域而龙肘山属金沙江流域。本研究结合植物区系研究和生物多样性研究，对该区的植物资源进行调查。通过样带调查和样线踏查结合，大量详实的野外样方调查和标本采集，进行传统的区系研究和生物多样性研究。研究该区物种多样性的海拔梯度格局及其潜在的影响影子，并利用新的区系评估质量方法对九顶山西坡的植物区系质量进行定量的研究，以期能更为深刻的理解该区的植物资源，为该区的资源保护和利用提供合理可行的建议。主要研究结论如下： 1）九顶山西坡植物区系的性质和特点 经鉴定和统计，九顶山西坡共有1707 种维管植物，分属617 属和140 科,其中种子植物1616 种，分属572 属117 科。就科的分布区成分构成而言，该区系的热带成分与温带成分相当，热带成分略占优势，表明九顶山西坡的植物区系与热带植物区系有较强的联系。但是，在九顶山西坡属的分布区类型所占的比例上，温带成分远远超过了热带成分，本区的种子植物分布表现出明显的温带性质。且温带分布类型的许多物种组成了九顶山西坡植被的建群种和优势种，是本区系最重要的成分，充分体现了本区系的温带性质。 2）九顶山西坡不同植被带的生物多样性海拔梯度格局 基于对土门-断头崖、茶山-九顶山、雁门沟-光光山三条垂直植被样带的调查，我们发现九顶山西坡的生物多样性沿海拔梯度的变化呈现出一定的规律性，不同样带之间有一定差异。就三条样带的物种组成相似性来看，虽然土门－断头崖样带属于涪江水系，而茶山－九顶山样带和雁门沟－光光山样带属于岷江水系，但不同水系对该区物种组成的影响并不明显。三条样带中，草本层物种丰富度均远远大于灌木层和乔木层，而以乔木层物种丰富度最低；α－多样性指数随着海拔梯度的变化在土门-断头崖样带中呈现单一下降趋势，在茶山-九顶山样带表现为双峰模型，而在雁门沟-光光山样带则表现为不显著波动变化；均匀度指数在土门-断头崖样带呈现出单一下降的趋势，在雁门沟-光光山样带表现为凹形曲线，而在茶山-九顶山样带却无明显的变化规律。β－多样性指数在土门-断头崖样带和茶山-九顶山样带呈现出明显的波动状态，植被类型替代现象明显；而在雁门沟-光光山样带却并未有十分显著的转折点，因其水平植被带受到干扰，同海拔替代现象不显著。 3）九顶山西坡维管植物丰富度的海拔梯度格局 我们考察了九顶山西坡和两条垂直样带（土门－断头崖和雁门沟－光光山样带）的不同分类等级（包括科、属、种）和不同生活型物种（乔木、灌木、禾草、蕨类和其它草本）的丰富度沿着海拔梯度的分布。结果发现，物种的丰富度海拔梯度格局具有不同的模式，单一下降和中间膨胀格局依然是其主流。不同生活型的物种具有不同的丰富度格局，但是对于环境需求相似的类型具有较相似的丰富度格局。不同的丰富度格局可能由多因素导致，包括：气候，海拔跨度，面积，人为干扰等等。 4）九顶山西坡区系质量评估 我们尝试使用传统的区系质量评估方法对九顶山西坡的区系质量进行评估，并尝试使用一种新的区系质量评估体系对该区的区系进行评价。在九顶山西坡随着海拔梯度的上升，平均保守性系数在各条植被带中均呈现出逐渐上升的趋势。区系质量指数随着海拔的升高都表现为双峰模型，在植被交错区区系质量指数相对较高，而在海拔的两极，区系质量指数都很低。大部分地区使用新方法计算所得的加权平均保守性系数和区系质量指数都比传统方法计算的平均保收性系数和区系质量指数要高，说明在九顶山西坡的三条样带中，大部分地区都是那些保守性系数较高的物种占据优势，同时也表明九顶山西坡具有很高质量的区系和自然植被。 5）龙肘山种子植物区系的性质和特点 龙肘山种子植物区系的物种较为丰富，共有154 科，544 属，1156 种。科的优势十分明显，单种属和寡种属数量众多，说明本区系植物成分较为复杂、起源古老、物种多样性指数较高。地理成分复杂，分布类型多样，其中热带成分在总数量上高于温带成分，但是许多温带成分的属是该区植被的重要建群类群和优势类群，表现出明显的亚热带性质。 6）龙肘山生物多样性的现状和特点 在海拔梯度上，龙肘山地区无论是科、属、种的数量，还是不同等级分类单元之间的数量比，均呈现先升后降的趋势，并在中海拔地区达到峰值。物种多样性指数从总体上来说变化幅度不大，略有先升后降的趋势，在中海拔梯度物种多样性最高。乔、灌、草三层的多样性指数表现出乔木层<灌木层<草本层的特征；乔木层均匀度的变化很大，而灌木层和草本层均匀度的变化较小；灌木层均匀度的波动又强于草本层。β－多样性指数呈现单峰模式，中海拔地区最高。就龙肘山东、西坡物种多样性相比较而言，两者虽然在数值上交替上升，但是却体现出了较为一致的趋势，但西坡因受到干热河谷气候的影响，其平均气温要高于东坡，导致了东坡植物群落和物种的分布比西坡要低。在区系成分构成上，低山区的相同海拔段，西坡的热带亚热带成分所占的比例要比东坡高，这是因为西坡的平均气温比东坡稍高，导致了热带、亚热带物种分布更多。而随着海拔的上升，东、西两坡的气候、土壤等条件趋于一致，其植物区系成分的构成格局也趋于一致。 The Hengduan Mountain region is a very natural floristic region; it belongs toChina-Himalaya sub-region of Holarctic region in floristic subarea of China. The flora in this areais rich in family, genus and species; has a very complex composition of geographical elements;especially with high richness of endemic species and obvious substitution phenomenon. Thisregion as a hot-spot area of floristic and biodiversity, has fascinated biologists in the world for along time. The eastern range of Hengduan Mountain is the transition zone of China – Himalayaforest sub-region and China-Japan forest sub-region in floristic. The water systems are quitedifferent, Minjiang River in the north and Jishajiang River in the south grow quit different but alsoabundant plant species and vegetation resources. The similarity and correlativity of Minjiang River dry valleys and Jinshajiang River dry valleys have provided advantageous condition tocontrast flora and biodiversity between north and south. In the present study, the Jiuding Mountainlies in the north of Hengduan Mountain and belongs to Minjiang River, and the LongzhouMountain lies in the south of Hengduan Mountain and belongs to Jinshajiang River. In our study, we combined the methods of floristic research and biodiversity investigation toexplore the resources of plant species and vegetations; sampled with transects along the altitudinalgradients and also with transverse straps with similar elevation; collected the vascular plant specimen with sampling plots of ecology. We explored the plant species richness patterns alongaltitudinal gradients and discussed the underlying factors aroused these patterns; and used a novelmethod to assess the quality of Jiuding Mountain’s flora. All for a deeper comprehension of the plant recourses of this region; and provided feasible and reasonable suggestion for the protectionof resources. The results were as follows: 1 The characteristic of the flora of the west slope of Jiuding Mountain We had collected 1707 species of vascular plants belonging to 617 genera in 140 families inthe west slope of Jiuding Mountain，in which included 1616 seed plant species belonging to 572genera and 117 families. As for the composition of the areal types of the Families of seed plants,tropic components and temperate components are well-balanced, and percentage of tropicscomponents is higher than that of temperate ones for a litter bit. This shows the flora in the westslope of Jiuding Mountain has strong relationship with the tropic flora. But for the composition ofthe areal types of genera, temperate components have far exceeded the tropics ones, indicated thewhole flora with a conspicuous temperate character. Temperate components possess maximumproportion in the west slope of Jiuding Mountain, and many of them belong to constructivespecies and dominant species in the vegetation, are most important components in JiudingMountain’s Flora, also have embodied the temperate character of this area sufficiently. 2 Biodiversity patterns along altitudinal gradients in different vegetation transects in the westslope of Jiuding Mountain Based on the investigation of three vegetation transects (including Tumen-Duantouya transect,Chashan-Jiudingshan transect and Yanmengou-Guangguangshan Transect) in the west slope ofJiuding Mountain, we found the change of biodiversity along the altitude gradients displayedcertain regularity, but have differences among different transects. The three transects belong todifferent water systems; the Tumen-Duantouya transect belongs to Fujiang River, and the othertwo belong to Minjiang River. From the similarity of species compositions of different transects,we found different water system didn’t show obvious impact on the species composition. In all thethree transects, the species richness of herb layer was remarkably higher than shrub and tree layer,and the species richness of tree layer was the lowest one. With the increasing of the altitude, theline of α-diversity was monotonically decreasing curve in Tumen-Duantouya transect, andbimodal curve in Chashan-Jiudingshan transect, but in Yanmengou-Guangguangshan transectshowed a wave-like curve although not very obvious. Species evenness showed monotonicallydecreasing trends in Tumen-Duantouya transect, and very low at mid-altitude in Yanmengou-Guangguangshan transect, but in Chashan-Jiudingshan transect changed irregularly. Changes inβ-diversity corresponded with the transition of vegetation in the Tumen-Duantouya transect andChashan-Jiudingshan transect, and the curve of β-diversity along altitude had obvious turningpoint; but in Yanmengou-Guangguangshan transect had no obvious turning point, and thesubstitution phenomenon was not obvious, transverse vegetation straps distributed interlaced. 3 Richness patterns of vascular plant species along altitude in the west slope of Jiuding Mountain Direct gradient analysis and regression methods were used to describe the species richnesspatterns along the altitudinal for Mt. Jiuding, as well as separately for Tumen-Duantouya Transectand Yanmengou-Guangguangshan Transect. Altitudinal gradient of diversity of units at differenttaxonomic level (including Family, Genus and Species) and at different life form (including tree,shrub, pteridophyte, grass and other herb) were tested to find differences among the richnesspattern. We found altitudinal richness also shows different patterns, and both monotonicallydecreasing pattern and hump-shaped pattern can be founded in vascular species richness. Speciesin different life forms show different altitudinal patterns, but those species with similarrequirements to environmental conditions show similar richness patterns along altitudinalgradients. Different richness patterns can be aroused by different climate, different altitudinal span,area factor, anthropogenic factor and so on. 4 Floristic quality assessments in the west slope of Jiuding Mountain We used both the conventional method broadly adopted in the USA and the new one toassess the floristic quality in the west slope of Jiuding Mountain. The Mean Coefficient ofConservatism (MC) had the trend of increment along the altitudinal gradients. The FloristicQuality Index (FQI) was a bimodal curve with increasing of elevation; FQI got maximum valuesin the transition zones of different vegetations in the middle altitude, and had very low values atthe two end of elevation. In most areas of the west slope of the Jiuding Mountain, the resultscalculated using the new methods were higher than those using the conventional method. Thisindicated the dominant species of the communities had very high coefficients of conservatism inmost areas of Jiuding Mountain, and the communities are relatively kept pristine and the habitats very integrative. 5 The characteristic of the flora of Longzhou Mountain The flora of Longzhou Mountain has very abundant in species composition; there are about1156 species of seed plants belonging to 544 genera in 154 families. In which, twelve families with more than 20 species include totally 232 genera and 532 species, and form the majority of itsflora. The origin of its flora is old, monospecific genera and oligotypic genera amounts to 510 innumber, which constitute 93.75% of total number of genera. The geographical components arevarious in Longzhou Mountain, the majority of flora are temperate and pantropic ones. The tropiccomponents overtopped temperate components on genera quantity, but many temperatecomponents belong to constructive species and dominant species in the vegetation, and the wholeflora shows an obvious subtropical character. 6 Current situation and characteristic of biodiversity in Longzhou Mountain With the increasing of altitude, the number of species, genus, family and the ratios ofdifferent taxonomic levels all displayed a trend of descending after rising first, and peaked atmiddle height area. The change of α-diversity was not very acutely, with the trend of descendingafter rising first in some degree, the middle height area had highest α-diversity. As studying thetree layer, shrub layer and herb layer respectively, the Shannon-Wiener index was in followingorder: tree layer < shrub layer < herb layer; the change of evenness was more complicatedly thanthat of diversity, the tree layer changed acutely, but the shrub layer and herb layer fluctuatedsmoothly. Changes in β-diversity also showed the trend of descending after rising first. TheJaccard index and Cody index all peaked at the middle height forest area. As for the comparison ofplant diversity and evenness between the west and east slope, the numerical values ascendedalternatively, but the trend of changing was similar. The distribution of similar plant communitiesand species in east slope were lower than the west slope for the influence of Jinsha River DryValley. As for the composition of different floristic components, in lower altitude area of westslope, the tropic and sub-tropic plants had higher ratio than east slope’s and even could be equal tothe temperate plants. With the increasing of elevation, the floristic composition become morelikely between the east and west slope and temperate plants dominated the flora.

川牛膝多糖的免疫活性及其水解活性片段分离研究

川牛膝多糖（CP）是从传统中药川牛膝（Cyathula officinalis Kuan）中提取的一种活性多糖，现代药理研究表明川牛膝多糖是川牛膝许多生物活性的物质基础。本实验室前期进行了川牛膝多糖的提取、分离、结构鉴定及其部分活性研究，发现川牛膝中多糖含量非常高，在对川牛膝多糖活性的初步研究中也证实了其具有免疫调节作用。我们为了进一步了解其免疫调节活性，并为构效关系的研究奠定基础，对其进行了如下研究： 1． 通过体外毒性检测、淋巴细胞增殖实验、NK细胞杀伤活性和腹腔巨噬细胞吞噬中性红活性测定，发现川牛膝多糖在10～300μg/mL浓度范围内，对细胞无毒性作用；能够促进LPS诱导的B淋巴细胞增殖(P<0.01)、增强NK细胞杀伤活性(P<0.05)和PMΦ吞噬中性红活性(P<0.01)，且随多糖浓度增高而增强；但其对ConA诱导的T淋巴细胞的增殖无促进作用(P>0.05)。 2． 通过正常小鼠体内淋巴细胞转化实验、迟发型变态反应分析、抗体生成细胞检测、碳粒廓清检测、腹腔巨噬细胞吞噬鸡红细胞活性和NK细胞活性测定，发现川牛膝多糖在适应性免疫方面能够促进SRBC免疫小鼠体内的抗体生成细胞的生成（P<0.01）和增强DNFB诱导的DTH（P<0.05），但对ConA诱导的脾淋巴细胞增殖无促进作用（P>0.05）；在固有免疫方面能够提高小鼠碳粒廓清速率（P<0.05），PMΦ吞噬 CRBC 活性（P<0.01）和NK细胞杀伤活性（P<0.05）。同时还发现其对由环磷酰胺(Cy)引起的白细胞数下降具有很好的抑制作用（P<0.01）。 3． 为了获得结构明确、均一的保留活性的川牛膝多糖片段，为其作用机制、构效关系研究提供关键研究材料，我们开展了“保留免疫活性的最小片段”的分离制备的初步研究。建立并优化了川牛膝多糖的酸水解条件，发现在6%的样品浓度，0.025mol/L的硫酸浓度，65℃的水解温度，水解时间为8min的条件下可以得到一系列连续的多糖片段；采用Bio-Gel P2 分子筛柱层析分离得到5个级分，通过体外淋巴细胞增殖实验、NK细胞活性测定、腹腔巨噬细胞吞噬中性红实验发现其中的一个片段仍保留较强的免疫活性，并测得其分子量约为2057Da，为保留免疫活性的最小片段的进一步分离奠定了基础。 Cyathula officinalis Kuan is a commonly-used Traditional Chinese Medicine (TCM) with a wide range of pharmacological activities. Modern pharmacological researches showed the polysaccharide extracted from it (CP) is an important component for many bioactivities of this TCM. In the previous studies, we found CP showed significant immuno-regulative activities. In order to evaluate this activity systematically and lay foundations for revealling its immuno-regulative machanisms and the Structure -Function relationship, we carried out the following research works: 1． The in vitro immunoactivities of CP were evaluated by using normal mice immunocytes with respects to cytotoxicity, lymphocytes proliferation, NK activity and the ability of peritoneal macrophage phagocytizing neutral red. The polysaccharide showed no cytotoxicity below the concentration of 300 μg/mL, and could promote B lymphocytes proliferation (P<0.01), enhance NK activity (P<0.05) and the ability of peritoneal macrophage phagocytizing neutral red (P<0.01) at the concentration of 10-300 μg/mL. The above effects were positively correlated with the concentration of the polysaccharides. But it could not promote T lymphocytes proliferation (P>0.05). 2． The in vivo immunoactivities of CP were observed on normal mice through the following indices: splenic lymphocyte transformation efficiency, delayed-type allergy, antibody-forming cells activity (AFC), rate of carbon clearance, rate of peritoneal macrophage phagocytizing chicken red blood cell (CRBC) and NK activity, and its influence on the decline of the mouse leucocyte count induced by Cy. The polysaccharide at medium-dose enhanced delayed-type allergy （P<0.05）and NK activity（P<0.05） and increased the rate of carbon clearance（P<0.05）, AFC activity（P<0.01） and the rate of peritoneal macrophage phagocytizing CRBC（P<0.01）. The polysaccharides also effectively resisted the decline of the mouse leucocyte count induced by Cy（P<0.01）. However, it couldn’t increase the splenic lymphocyte transformation efficiency（P>0.05）. 3． Attempting to isolate and prepare the minimal fragments retaining activity with identical structure for further studying on immuno-regulative mechanism and Structure-Function relationship, we carried out the study on hydrolysis of CP, isolation of hydrolysed fragments, and the activity evaluation of the isolated fragments. CP with concentration of 6% was hydrolysed at 65℃ for 8 min with sulfuric acid of 0.025 mol/L，then the hydrolysate was separated using Bio-Gel P2 chromatography, 5 portions of fragments were obtained. The immunoactivities of these fragments were evaluated by using normal mice immunocytes with respect to lymphocytes proliferation, NK activity and ability of peritoneal macrophage phagocytizing neutral red. One fragment with relative molecular mass of 2057Da was found retaining immunoactivity.