燕麦嗜酸菌鞭毛素的提纯及其活性检测

为阐明燕麦嗜酸菌(Aaa)鞭毛素作为一种病原相关分子模式(PAMP)诱导水稻免疫反应(PTI)的作用机理,采用机械震荡、丙酮沉淀以及离子交换层析进行了蛋白提纯,并用DAB染色法对其诱导水稻产生H2O2的能力进行了测定。结果表明,上述提纯方法的组合使用有效地提纯了分子量约为45 kD的鞭毛素,经浸润接种处理后诱导了水稻叶组织H2O2的产生。因此,Aaa鞭毛素提纯蛋白可用于后续的水稻PTI机理分析。     

一种快速准确测定A群脑膜炎奈瑟菌活菌率的方法

目的研究"美蓝-(碱性)藏红O"双染色法用于快速准确测定A群脑膜炎奈瑟菌(group A Neisseria meningitidis,以下简称A群脑膜炎球菌)活菌率的可行性。方法制备A群脑膜炎球菌悬液,取菌悬液双染色法染色,用光学显微镜观察,计算活菌率。将菌悬液进行10倍系列稀释,各菌液分别用平板涂布法接种至10%羊血普通琼脂平皿,于CO2环境下、35~37℃培养12~20 h,计算菌悬液活菌率。比较两种方法测定8批A群脑膜炎球菌所测得的活菌率,并进行相关性和线性回归分析。结果 "美蓝-(碱性)藏红O"双染色法与平板涂布法所测得的活菌率结果差异无统计学意义(P>0.05);两种方法所测得的活菌率具有显著的直线回归关系(P<0.01,n=8),相关系数r=0.984,其回归方程为y=0.919x+1.661(R2=0.967)。结论 "美蓝-(碱性)藏红O"双染色法检测时间短、判定结果直观、操作简单、计数结果准确,可用于A群脑膜炎球菌活菌率的快速测定。     

实验动物整体血管和心脏冰冻切片的脂质染色法改进与应用

在实验动物脂质代谢的模型建立中,所进行的Sundon(苏丹)染色效果较差,而经过改进的Oil red O (油红O)染色法,能够使整体胸、腹主动脉血管和心脏冰冻切片的脂质沉着物色彩鲜艳、对比清晰、方法可靠,是较为理想的脂质染色方法.     

短暂缺氧致新生大鼠海马内β-连环素、神经源性分化因子1和胰岛素受体表达动态改变

目的 探讨短暂缺氧对新生大鼠海马β-连环素(β-catenin)、神经源性分化因子1(NeuroD1)、胰岛素受体(insulin receptor)表达的影响.方法 将3日龄SD大鼠置于8％O2+92％N2的混合气体中2h,采用尼氏染色及透射电镜观察海马结构病理变化;于缺氧后2h、4h、24h断头取脑,免疫荧光染色法...     

淋病诊断试剂盒的研究

本文叙述了试剂盒的制备和考核的全过程,试剂盒用6个国际标准株、46个淋球菌临床株、32株不动杆菌和51株非淋球菌临床株对其敏感性、特异性进行了考核,并同革兰氏染色法和细菌培养法作了平行对比试验,结果表明本试剂盒具有敏感(O.3亿/ml)、特异(100%)、快速(3min)的特点.三种方法的比较研究证明革兰氏染色法的准确率为95.6%,培养法和试剂盒的准确率均为100%;与培养法的符合率革兰氏染色法为95.6%,试剂盒为100%.     

三氧化二砷对K562细胞凋亡的诱导及生长抑制作用的研究

目的：研究三氧化二砷(As2O3)对人红白血病细胞株K562的生长抑制和凋亡诱导作用。方法：以As2O3作为耐药逆转剂,用台盼兰排染法,噻唑兰(MTT)还原法,Hoechst 33342和PI荧光染色法,流式细胞仪技术和荧光分光光度法,观察了不同浓度的As2O3(0．2—5．0μmol/L)对人红白血病细胞株K562的生长抑制和凋亡诱导作用。结果：As2O3对K562细胞具有明显生长抑制和凋亡诱导作用,其作用强度在一定范围内均具药物浓度和时间依赖性。结论：As2O3主要以诱导肿瘤细胞凋亡而表现其毒性作用。     

芝加哥天蓝法在甲屑真菌检测中效果的评估

目的探讨芝加哥天蓝染色法对甲真菌病患者标本检测效果。方法收集皮肤科门诊确诊的甲真菌感染患者标本160例,同时用KOH湿片法、荧光染色法和芝加哥天蓝染色法对真菌进行镜下形态学观察,并比较真菌检出率。结果芝加哥天蓝染色法和荧光染色法对真菌的形态结构清晰易辨。芝加哥天蓝染色法、荧光染色法和KOH湿片法的检出率分别为60.6%、68.5%、47.5%;荧光染色法与芝加哥天蓝染色法的检出率结果比较差异无统计学意义(χ~2=0.176 2,P=0.674 0),芝加哥天蓝染色法和荧光染色法与KOH湿片法的结果比较差异有统计学意义(χ~2=102.012 9,P0.000 1;χ~2=57.010 3,P0.000 1)。结论如实验室没有荧光显微镜,芝加哥天蓝染色法检测甲真菌病患者标本效果较好。     

箱式通量观测技术和方法的理论假设及其应用进展

碳(CO_2、CH_4)、氮(N_2O)和水汽(H_2O)等温室气体的交换通量是生态系统物质循环的核心,是地圈-生物圈-大气圈相互作用的纽带。稳定同位素光谱和质谱技术和方法的进步使碳稳定同位素比值(δ~(13)C)和氧稳定同位素比值(δ~(18)O)(CO_2)、δ~(13)C(CH_4)、氮稳定同位素比值(δ~(15)N)和δ~(18)O (N_2O)、氢稳定同位素比值(δD)和δ~(18)O (H_2O)的观测成为可能,与箱式通量观测技术和方法结合可以实现土壤、植物乃至生态系统尺度温室气体及其同位素通量观测研究。该综述以CO_2及其δ~(13)C通量的箱式观测技术和方法为例,概述了箱式通量观测系统的基本原理及分类,阐述了系统设计的理论要求和假设,综述了从野外到室内土壤、植物叶-茎-根以及生态系统尺度箱式通量观测研究的应用进展及问题,展望了气体分析精度和准确度、观测数据精度和准确度以及观测数据的代表性评价在箱式通量观测研究中的重要性。     

Nano-SiO_2对子代雌性小鼠动情周期的影响

目的:探讨纳米二氧化硅(Nano-Si O2)对子代雌性小鼠动情周期的影响。方法:选择8周龄的ICR孕鼠6只,随机平均分为两组,一组在E5.5~E11.5时隔天注射纳米二氧化硅,另外一组注射PBS,每组取子代雌性小鼠各3只,待其自然生长到7周龄时,每日采用巴氏染色法检查其动情周期。结果:与注射PBS的子代雌性小鼠相比,注射Nano-Si O2的子代雌性小鼠的动情周期的总时间无明显差异(P0.05),动情前期的时间明显缩短,动情后期的时间明显延长(P0.05),动情期虽然缩短,但差异不显著(P0.05)。结论:孕期暴露于Nano-Si O2未导致子代雌性小鼠动情周期的紊乱,但对其生殖功能可能有一定的影响。     

六种杜鹃花属植物花粉活力测定方法的比较研究

选取睫毛杜鹃(Rhododendron ciliatum)、多鳞杜鹃(Rhododendron polylepis)、薄皮杜鹃(Rhdodenron taronense)、映山红(Rhododendron simsii)、马银花(Rhododendron ovatum)和比利时杜鹃(Rhododendron hybridum)为研究对象,通过蔗糖、H3BO3、CaCl2单因子及L25( 53)正交试验对它们进行花粉萌发试验研究,比较I2-Kl染色法、TTC染色法、联苯胺染色法、MTT染色法4种花粉活力测定方法的不同.结果表明:蔗糖、H3BO3、CaCl2及3因子交互效应对杜鹃花花粉萌发有显著影响.适宜的离体培养基配方依杜鹃花种类不同而不同,睫毛杜鹃为:蔗糖150 g/L +H3BO3 0 mg/L +CaCl2 50 mg/L;映山红为:蔗糖100 g/L +H3BO3 100 mg/L +CaCl20 mg/L;马银花为:蔗糖50 g/L+ H3BO3 200 mg/L +CaCl2 0 mg/L;比利时杜鹃为:蔗糖150 g/L+ H3BO3100 mg/L +CaCl2 0 mg/L.MTT染色法是简单快速测定杜鹃花花粉活力的最适染色法.     

Sequential maturation of skeletal muscle groups during postnatal ontogenesis of rats]

Experiments were conducted on rats during the early postnatal period; a study was made of the membrane potential (MP) establishment of the fibers of the skeletal muscles of the neck, the anterior and the posterior limbs. At birth the most mature were the muscles of the neck, and the least -- the muscles of the posterior limb. Establishment of the stationary MP level in the muscles of the neck occurred during the first week after birth, in the muscles of the anterior limbs -- by the 10th-12th day, and of the posterior limbs -- by the 15th-20th day. The order of maturation of various groups of the skeletal muscles was associated with the peculiarities of the neuro-trophic influences at various age periods. Muscles of the neck were characterized at all the developmental stages by a rhythmic low-frequency electromyographic activity. In the muscles of the limbs the rhythmic electromyographic activity was transformed into the discharge high-frequency activity by the period of termination of increase of the MP of the muscles.     

Dependence of anticipatory changes in the hand muscle activity and the grip force on the height of the fall when catching a falling object

The time course of changes in the hand muscle activity and the grip force before the hit of an object falling from different heights into a cup held between the thumb and the forefinger was analyzed in three variants of the experiment: (1) the subject saw the object falling; (2) the subject did not see the object falling but initiated the fall; and (3) the subject had no information on either the falling or its start. In the third variant, the muscle activity and the grip force changed in response to the object hitting the cup. In the second variant, the muscle activity and the grip force began to change 200–280 ms before the hit, this time being independent of height from which the object fell. In the first variant, the anticipatory changes began 150 ms after the object started falling and did not depend on the height of the falling within the rage 30–50 cm. If the object fell from a height of 70–105 cm, the changes in the muscle activity and the grip force began a fixed time before the object hit the cup, which did not depend on the height from which the object fell. Thus, when the object fall from small heights, the timing of the increase in the grip force was mainly determined by the moment when the object began moving; at large heights, the increase in the grip force was related to the presumed moment of the hit.     

Ab initio molecular orbital study of the interaction of Li, Na and K with the pore components of ion channels: Consideration of the size, structure and selectivity of the pore of the channels

Ab initio molecular orbital calculations were made for the various types of structures of the pore of the ion channel and the results were applied to the permeability model by Hille, an extension of the Eyring rate theory. In Hille's model, ion passage through the channel is regarded as a kinetic process. Accordingly, it is thought that the interaction energy between cation and ligand, the easier the passage is, due to the lower activation energy. The calculated interaction energy was in the order Li+ greater than Na+ greater than K+ for all models. The optimum size of the pore determined from the interaction energy depends on the structure of the filter. The size for the pentagon was largest, followed by the hexagon and tetragon. On the other hand, the size depends hardly at all on the kind of ligand molecules. In the case of the tetragon, the sizes for the Na and K channels were nearly the same as those estimated from the model building and inhibitor-blocking experiment. The interaction energy between the ionized carboxyl group and the cation was extremely large, clearly reflecting the experimental fact that the carboxyl group in the pore has an important role in making the passage of the cation through the channel easier by dehydrating the water molecules. By analysis of the interaction energy, it was revealed that the contribution of the electrostatic energy was predominant, although the contributions of the other effects might not be negligible. Among these effects, the value of the charge transfer energy is largest, and this is noteworthy in connection with the selective transmission of cations through the overlap of orbitals. It is concluded that the quantum-chemical indices such as interaction energy and the electronic charge calculated by the sophisticated ab initio method help to shed light on the nature of the pore of the ion channel.     

The protein arrestin as a regulator of the phototransduction process and as a factor of pathogenesis of the eye diseases

In the first part of the paper, the results of the investigation of the rhodopsin arrestin interaction are presented. The results were mainly obtained with the technique of the selective labelling of the rhodopsin and arrestin SH-groups and the rhodopsin limited proteolysis. These results are discussed in the frame of the latest data on the three-dimensional structure of arrestin. In the second part of the paper, results of the antigenic properties of arrestin (S-antigen) and its role in the pathogenesis of the retina diseases are summarized. The data on the role of the autoimmune processes in the pathogenesis of diabetic retinopathy are presented. We have also described the results of the use of the elaborated technique of the immune diagnostics in the prognosis of the diabetic retinopathy and retinopathy of the premature babies.     

纵坑切梢小蠹蛀梢期空间分布

在昆明地区,纵坑切梢小蠹（Ｔｏｍｉｃｕｓ　ｐｉｎｉｐｅｒｄａ）成虫蛀梢多集中在蛀干木附近。种群密度以蛀干木为中心向周围是指数递减,散布半径约３０ｍ。在蛀虹梢过程中,该种群逐渐向新区扩张。在树冠内,纵坑切梢小蠹主要分布在４—１０轮枝上。第７轮技虫口百分率最高。６—７轮枝受害率最大。树冠上层受害较其下属于重。从树冠水平层次考察,树冠外层虫量相对集中,约为树冠中、内层虫量之和。树冠内层虫量最少。纵坑切梢小蠹在树冠内的种群分布系由梢径、种群密度、蛀梢行为、降落方式、光照等因素综合影响的结果。     

Kinetic characterization of the substrate specificity and mechanism of mushroom tyrosinase.

This paper reports a quantitative study of the effect of ring substituents in the 1-position of the aromatic ring on the rate of monophenol hydroxylation and o-diphenol oxidation catalyzed by tyrosinase. A possible correlation between the electron density of the carbon atom supporting the oxygen from the monophenolic hydroxyl group and the V Mmax values for each monophenol was found. In the case of o-diphenols the same effect was observed but the size of the side-chain became very important. NMR studies on the monophenols justified the sequence of the V Mmax values obtained. As regards the o-diphenols, on the other hand, only a fair correlation between NMR and V Dmax values was observed due to the effect of the molecular size of the ring substituent. From these data, it can be concluded that the redox step (k33) is not the rate-determining step of the reaction mechanism. Thus, the monophenols are converted into diphenols, but the order of specificities towards monophenols is different to that of o-diphenols. The rate-limiting step of the monophenolase activity could be the nucleophilic attack (k51) of the oxygen atom of the hydroxyl group on the copper atoms of the active site of the enzyme. This step could also be similar to or have a lower rate of attack than the electrophilic attack (k52) of the oxygen atom of the active site of oxytyrosinase on the C-3 of the monophenolic ring. However, the rate-limiting step in the diphenolase activity of tyrosinase could be related to both the nucleophilic power of the oxygen atom belonging to the hydroxyl group at the carbon atom in the 3-position (k32) and to the size of the substituent side-chain. On the basis of the results obtained, kinetic and structural models describing the monophenolase and diphenolase reaction mechanisms for tyrosinase are proposed.     

Interrelationships between serum and secretory antibodies in immunization with live influenza vaccine]

A study was made of the accumulation of antibodies in the blood serum and the secretions of the respiratory tracts of persons immunized with the living influenza vaccine. The duration of inductive phase and the dynamics of the antibody accumulation in the secretions occurred irrespective of their initial level in the blood sera, this pointing to the autonomic character of the local immunity system. On the other hand the functional condition of the system of local immunity influenced the intensity of the antibody formation in the system of the general immunity. If before the immunization the antibody titre in the secretions were 1:4 and greater, the antibody accumulation in the blood sera took place less intensively. An analogous phenomenon was also observed when the antibodies were absent in the secretions before the immunization, but their formation took place as soon as the first week after it. The mechanism of this peculiar "competition for the antigen" of the systems of local and general immunity consisted in the neutralization of the influenza virus in the area of the porta of infection.     

Comparison of the power and accuracy of biallelic and microsatellite markers in population-based gene-mapping methods.

Because of their great abundance and amenability to fully automated genotyping, single-nucleotide polymorphisms (SNPs) and simple insertion/deletion are emerging as a new generation of markers for positional cloning. Although the efficiency and cost associated with the markers are important in the mapping of human disease genes, the power to detect the linkage between the marker and the disease locus, as well as the accuracy of the estimation of the map location of the disease gene, dictate the selection of the markers. Both the power and the accuracy depend not only on the type of the markers but also on other factors, such as the age of the disease mutation, the magnitude of the genetic effect, the marker-allele distribution in the population, mutation rates of marker loci, the frequency of the disease allele, the recombination fraction, and the methods for mapping the human disease genes. In this article, we develop a mathematical framework and the analytical formulas for calculation of the power and the accuracy and investigate the impact that the aforementioned factors have on the power and the accuracy, by using two population-based gene-mapping methods-likelihood-based linkage-disequilibrium mapping and the transmission/disequilibrium test, for both biallelic SNPs and microsatellites. These studies provide not only guidance in selection of the markers and in the design of the sample scheme for positional cloning but also insight into the biological bases of the mapping of human disease genes.     

Effects of transections and electrical coagulations in the medulla oblongata upon the activities in the respiratory muscles of the crucian carp (author's transl)]

The following conclusions may be drawn from the results in this work. The respiratory cycles are formed by the neuronal machinery in the reticular formation under the posterior part of the vagal motor nucleus. The motor neurones or the neuronal networks composing the motor nucleus of the respiratory muscles tonically discharge the action potentials, when the neurones or the networks are released from the inhibitory influences of the interneurones connecting the neuronal machinery to the motor neurones. Furthermore, the interneurones probably generate the tonic discharges after removing the inhibitory influences of the other interneurones or the neuronal machinery on them. A reflex mouth closing is elicited by a mechanical stimulus applying on the upper lip. The motor neurones of the m. adductor mandibulae are activated via only one synapse in the reflex. The reflex action potentials recorded from the motor nerve reduce in amplitude at the resting phase of the nerve in the respiratory cycles. These results suggest that the respiratory motor neurones are by nature spontaneous generators of the tonic action potentials and, in the time of the normal breathing, the tonic activity is interrupted by an inhibitory influence of the neuronal machinery generating the respiratory cycles.     

银杏套细胞发育的解剖学研究

银杏套细胞起源于近雄配子体表达的颈卵器母细胞周围细胞,中央细胞形成时,套细胞呈明显的一层,围绕颈卵器紧密排列,随着颈卵器的发育,套细胞体积逐渐地大,细胞质变浓厚,细胞质中脂滴增多,套细胞和中央细胞的接触壁开始出现局部加厚,在颈卵器发育的泡沫化阶段,套细胞和中央细胞的接触壁不均匀加厚较为显著,在较薄的区域可见胞间连丝,受精前,套细胞中液泡增多,脂滴迅速减少,造淀粉体增大,套细胞与卵细胞的接触壁的不均匀加厚非常明显,精核进入颈卵器以后,卵细胞与套细胞的接触壁和卵细胞的质膜之间形成一个薄厚不均的间隔层,受精卵分裂时,受精卵细胞与套细胞接触壁的凹陷处可见许多小泡和内质网,游离核期时,套细胞内出现大量小液泡,细胞内含物迅速消失,套细胞外形变长,胚胎长出颈卵器后,套细胞逐渐解体消失。