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.

A pilot experiment for mass measurement at CSRe

A pilot experiment of mass measurement was performed at CSRe with the method of isochronous mass spectrometry. The secondary fragments produced via RIBLL2 with the primary beam of 400 MeV/u, Ar-36 delivered by CSRm were injected into CSRe. The revolution periods of the stored ions, which depend on the mass-to-charge ratios of the stored ions, were measured with a time-of-flight detector system. The results show that the mass resolution around 8 x 10(-6) for Delta m/m is achieved.

纳秒激光诱导C60分子碎裂中轻碎片离子C+n(n＜30)的产生机理研究

The time of flight mass spectrometric technique was used to determine the initial mean kinetic energy of small fragment ions C-n(+) (n <= 11) produced from C-60 excited by 532 nm nanosecond laser pulses. The measured kinetic energy shows little variation with the fragment mass and the laser fluence in a broad range. Based on the assumption that C-30(+) is produced predominantly by a single electron emission followed by successive C-2 evaporation from hot C-60 in the nanosecond laser field, the formation of small fragments is interpreted as the complete breakup of the unstable C-30(+) cage structure. The interpretation is consistent with the previously observed results.

beta-delayed fission of Ac-228

The basic process of an exotic decay mode namely P-delayed fission is simply introduced. The progress status of the study in the world is essentialized. The observation of P-delayed fission of Ac-228 is reported. The radium was radiochemically separated from natural thorium. Thin Ra sources in which Ac-228 was got through Ra-228 ->(beta-) Ac-228 were prepared for observing fission fragments from beta-delayed fission Ac-228. They exposed to the mica fission track detectors, and measured by an HPGe gamma-ray detector. The beta-delayed fission events of Ac-228 were observed and its beta-delayed fission probability was found to be (5 +/- 2) x 10(-12).

Studies on the exotic structure of Al-23

The longitudinal momentum distribution (P-//) of fragments after one-proton removal from Al-23 and reaction cross sections (sigma(R)) for Al-23,Al-24 on carbon target at 74A MeV have been measured simultaneously. An enhancement in sigma(R) is observed for Al-23 compaxed with Al-24. The full width at half maximum of the P-// distribution for Mg-22 fragments has been determined to be 232 +/- 28 MeV/c. Analysis of P-// using the Few-Body Glauber Model indicates a dominant d-wave configuration for the valence proton in the ground state of Al-23. The exotic structure in Al-23 is discussed.

Isospin effect of projectile fragment yields

Isospin asymmetry is very important in the nuclear equation of state (EOS), isotope yield from the projectile fragments can give information of the reaction process. In this paper projectile fragment yields are measured in the collision Ar-36,Ar-40 + Ni-64 at incident energy 50 MeV/u with different isospin asymmetry project Ar-36,Ar-40, data analysis, particle identification and event selection are described. Isotope yields are compared in these two reactions, and axe also compared with the empirical parametrization of fragmentation cross-section calculated by EPAX.

Biodosimetry estimate for high-LET irradiation

The purpose of this paper is to prepare for an easy and reliable biodosimeter protocol for radiation accidents involving high-linear energy transfer (LET) exposure. Human peripheral blood lymphocytes were irradiated using carbon ions (LET: 34.6 keV mu m(-1)), and the chromosome aberrations induced were analyzed using both a conventional colcemid block method and a calyculin A induced premature chromosome condensation (PCC) method. At a lower dose range (0-4 Gy), the measured dicentric (dics) and centric ring chromosomes (cRings) provided reasonable dose information. At higher doses (8 Gy), however, the frequency of dics and cRings was not suitable for dose estimation. Instead, we found that the number of Giemsa-stained drug-induced G2 prematurely condensed chromosomes (G2-PCC) can be used for dose estimation, since the total chromosome number (including fragments) was linearly correlated with radiation dose (r = 0.99). The ratio of the longest and the shortest chromosome length of the drug-induced G2-PCCs increased with radiation dose in a linear-quadratic manner (r = 0.96), which indicates that this ratio can also be used to estimate radiation doses. Obviously, it is easier to establish the dose response curve using the PCC technique than using the conventional metaphase chromosome method. It is assumed that combining the ratio of the longest and the shortest chromosome length with analysis of the total chromosome number might be a valuable tool for rapid and precise dose estimation for victims of radiation accidents.

Dependence of isoscaling parameters on nucleon-nucleon cross section and momentum-dependent interaction

Influences of the isospin dependence of the in-medium nucleon-nucleon cross section and the momentum-dependant interaction (MDI) on the isotope scaling are investigated by using the isospin-dependent quantum molecular dynamics model (IQMD). The results show that both the isospin dependence of the in-medium nucleon-nucleon cross section and the momentum-dependent interaction affect the isoscaling parameters appreciably and independently. The influence caused by the isospin dependence of two-body collision is relatively larger than that from the MDI in the mean field. Aiming at exploring the implication of isoscaling behaviour, which the statistical equilibrium in the reaction is reached, the statistical properties in the mass distribution and the kinetic energy distribution of the fragments simulated by IQMD are presented.

Entrance channel dependence of the correlation function of IMFs in Ar-36+Sn-112,Sn-124 at 35MeV/u

The reduced velocity correlation functions of the Intermediate Mass Fragments (IMFs) were measured in the reactions of Ar-36+ Sn-112,Sn-124 at 35MeV/u. The anti-correlation at small reduced velocities is more pronounced in Ar-36+ Sn-124 system than that in Ar-36+ Sn-112 system. The difference of the correlation functions between the two reactions is mainly contributed by the particle pairs with high momenta. A three-body Coulomb repulsive trajectory code (MENEKA) is employed to calculate the emission time scale of IMFs for-the both systems. The time scale is 150fm/c in the Ar-36+ Sn-112 system and 120fm/c in the Ar-36+ Sn-124 system, respectively. A calculation based on an Isospin dependence Quantum Molecular Dynamics code (IQMD) reveals that the emission time spectrum of IMFs is shifted slightly leftwards in Ar-36+ Sn-124 compared with that in the Ar-16+ Sn-112 system, indicating a shorter emission time scale. Correspondingly, the central density of the hot nuclei decreases faster in Ar-36+ Sn-124 than in Ar-36+ Sn-112

A transverse field gas ionization chamber performed at RIBLL

A transverse field gas ionization chamber as Delta E detector at the Radioactive Ion Beam Line in Lanzhou (RIBLL) is described. A high detection efficiency and long plateau are achieved with the mixed gas Ar(80%)+CO2(20%). The energy resolution is 3.25% for 4.94MeV alpha particle. This ionization chamber has been tested in the experiment with 50MeV/u Ni-58 bombarding Ta at RIBLL. All the fragments can be identified clearly by the ionization chamber.

alpha decay half-lives of new superheavy nuclei within a generalized liquid drop model

The alpha decay half-lives of the recently produced isotopes of the 112, 114, 116 and 118 nuclei and decay products have been calculated in the quasi-molecular shape path using the experimental Q(alpha) value and a Generalized Liquid Drop Model including the proximity effects between nucleons in the neck or the gap between the nascent fragments. Reasonable estimates are obtained for the observed alpha decay half-lives. The results are compared with calculations using the Density-Dependent M3Y effective interaction and the Viola-Seaborg-Sobiczewski formulae. Generalized Liquid Drop Model predictions are provided for the alpha decay half-lives of other superheavy nuclei using the Finite Range Droplet Model Q(alpha) and compared with the values derived from the VSS formulae.

Critical behavior of multifragmentation in finite nuclei and extraction of critical exponents

Using a phenomenological asymmetric nuclear equation of state, we obtained pressure-density isotherms of the finite nucleus Sn-112 simulated in r-space and in p-space and constructed the nuclear fragments by using the coalescence model. After correlatively analysing the fragments, the signal of critical behavior has been found and critical exponents were also extracted.

FIRST ISOCHRONOUS MASS MEASUREMENTS AT CSRe

With the commissioning of the Cooler Storage Ring at the Heavy Ion Research Facility in Lanzhou (HIRFL-CSR), a pilot experiment operating the CSRe in isochronous mode to test the power of HIRFL-CSR for measuring the mass of the short-lived nucleus was performed in December of 2007. The transition point gamma t of CSRe in isochronous mode is 1.395 which corresponds to the energy about 368 MeV/u for the ions with atomic number-to-charge ratio A/q = 2. The fragments with A/q = 2 of Ar-36 were injected into CSRe and their revolution frequencies were measured with a fast time pick-up detector with a thin foil in the circulating path of the ions. A mass resolution of better than 105 for m/Delta m was achieved.

DIRECT MASS MEASUREMENTS OF EXOTIC NUCLEI IN STORAGE RINGS

Mass measurements of exotic nuclei is a fast, developing field which is essential for basic nuclear physics and a wide range of applications. The method of storage ring mass spectrometry has many advantages: (1) a large amount of nuclides can be simultaneously measured; (2) very short-lived (T-1/2 greater than or similar to 50 mu s) and very rare species (yields down to single ions) can be accessed; (3) nuclides in several atomic charge states can be investigated, (4) half-life measurements can be performed with time-resolved mass spectrometry. In this contribution we concentrate on some recent achievements and future perspectives of the storage ring mass spectrometry.