Table of Content

20 March 2000, Volume 36 Issue 2

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Analysis of the Mouse Nodal Gene Promoter on Its 5' Flanking Sequence

HU Xinli,ZHOU Xunlei

2000, 36(2):  178-185. 

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Nodal gene is critical to mouse development. It plays a vital role in the mesoderm formation and also has a biological function during mouse embryo pattern formation. To determine the basal promoter, the 5' flanking sequence of Nodal gene were truncated and inserted into the luciferase reported vector. These constructs were transiently transfected into F9 cells and the luciferase activity of each construct was determined. The basal promoter was located at about 60bp up stream of the transcription initiation site. It contains a TATA box at -33bp which is required for the transcription initiation. This basal promoter is active in many cell types, but its activity is not very strong.

Some Preliminary Synthetic Studies Related to Natural Perylenequinones

FU Bin,ZHANG Pang

2000, 36(2):  167-171. 

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3-Allyl-6, 8-dimethoxy-1-tetralone as the intermediate projected for a synthesis of hypocrellin was studied for its facile formation from 3-(3, 5-dimethoxy-benzyl)-5-hexenoic acid and its Wacker oxidation, addition of bromine and of hydrogen chloride. It and the diketone obtained by its Wacker oxidation were shown not susceptible towards oxidation by selenium dioxide.

Preparation of PVA-g-NIPAAm Hydrogel by γ-Radiation and Its Controlled Release Research

LIU Ning,ZHAI Maolin,LI Jun,YI Min

2000, 36(2):  172-177. 

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Thermosensitive PVA-g-NIPAAm copolymers was prepared by grafting copolymerization of N-isopropylacrylamide (NIPAAm) onto poly(vinyl alcohol) (PVA) in homogeneous system by ⁶⁰Co-γ irradiation at room temperature. The effects of radiation grafting conditions, such as absorption dose, dose rate, acid concentration on grafting yield were investigated. It was found that dose rate and acid concentration had obvious influence on the grafting yield. Then the hydrogels of PVA and PVA-g-NIPAAm copolymer were made through freezing-thawing processes and the later hydrogels obtained in this method were found to exhibit obvious thermal sensitivity. In addition, the slow release of Methylene Blue (MB) from this kind of hydrogels was studied.

Studies on Polarographic Adsorptive Complex Wave of Indium(III)-lumogallion

BAO Xiaoyu,ZHU Zhiwei,LI Nanqiang

2000, 36(2):  149-152. 

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In (CH₂)₆N₄-HCl buffer solution at pH 5.5, a sensitive adsorptive complex wave of In(III)-lumogallion(LMG) was obtained by single-sweep polarography. The change of peak height with the concentration of indium is linear in the range from 1.0×10^-7mol/L to 1.0×10^-5mol/L. The detection limit is down to 8×10^-8mol/L. The composition of the complex was detected as In(III): LMG=1:2. The conditional stability constant was determined as β=3.6×10¹³. The rate constant of the surface electrochemical reaction was determined as k_s1=4.4s^-1, k_s2=1.74s^-1, k_s3=10.8s^-1.

Synthesis and Photophysical Properties of Cyano-Bridged Complexes of Ru(II)

LI Hongliang,GUO Guolin,GUI Linlin,LI Chongjia

2000, 36(2):  153-159. 

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The special property of the NO⁺ ligand which can be solvated by azide-assisted labilization of the nitrosyl ligand in the Ru(II) complexes was applied in the synthesis of the necomplexes: NO₂-Ru(bpy)₂-NC-Ru-(py)₄-CN⁺, Cl-Ru(bpy)₂-NC-Ru(py)₄-CN⁺, NO₂-Ru(bpy)₂-NC-Ru(py)₄-CN-Ru-(bpy)₂-NO₂²⁺ and Cl-Ru(bpy)₂-NC-Ru(py)₄-CN-Ru(bpy)₂Cl²⁺(bpy=2, 2'-bipyridine, py=pyridine). The UV-Visible spectra, the IR spectra and the photo-luminescence properties of these complexes were studied. It has been shown from the elemental analysis, the UV-Visible spectra and the IR spectra, that the new complexes we designed were prepared. It is very interesting that there are two different MLCT(metal-to-ligand charge transfer) process and two photo-luminescence emission bands in one molecule. These special photophysical properties were the results of the special configuration and buildups of the different blocks in the new complexes. This will present the advantage for the study of the electron transfer within those molecules.

Localization of Specific Glycoproteins of Sperm Cell Membrane from Zea mays by Electronmicroscopy Immuno-gold Labelling

YANG Zhonghan,CHEN Li,SHAO Linxiang,WANG Jie

2000, 36(2):  193-198. 

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Plasma membrane proteins of sperm cells from Zea mays were passed through affinity column, separated by SDS-PAGE and stained using Con A-HRP. The components of 37kD and 39kD glycoproteins from gel were immunized respectively mice and polyclonal antiserums were obtained. Glycoproteins of sperm cell surface can be examined using IgG(rabbit anti mouse)-colloidal gold(15nm) as probe. The results showed that distribution of 37 and 39kD peptides on the plasma membrane surface of sperm cells of maize with visible immunogold particles to be localized. We consider that 37 and 39kD polypeptide were specific plasma membrane glycopeptides of sperm cells. Research of their function is to be continued.

Cloning and Expression of the Rabbit Growth Hormone cDNA

DONG Yuqing,LI Ronggui,ZHAO Jindong,WU Guangyao

2000, 36(2):  199-202. 

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The cDNA of preprotein and mature protein of rabbit hypophysiscerebri was obtained. The cDNA of mature growth hormone was cloned in pET-3a, then the recombinant expression vector was transformed into E.coli BL21(DE3 plys). The overexpression was performed and the yield was 40% of the total protein.

Photoelectrochemical Studies on the SnO₂ Nanostructured Porous Film Sensitized by Cyanine Dye

ZHANG Li,YANG Maizhi,QIAO Xuebin,HAO Yanzhong,GAO Enqin,CAI Shengmin,MENG Fangshun,TIAN He

2000, 36(2):  160-166. 

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The photoelectrochemical behaviors of the SnO₂ nanostructured porous film sensitized by cyanine dye were investigated in this paper. The results showed that the excitation state level matched the conduction band edge of SnO₂ nanoparticle. Therefore the sensitization of this dye can increase the photocurrent intensity of SnO₂ nanostructured electrode obviously and results in a red-shift of optical absorption from the ultra-violet region to the visible region. As a result, the light-to-electricity conversion efficiency was improved evidently, the maximum value of IPCE has reached 28.9%.

A Study on the Development of the Anther and the Male Gametophyte in Cimicifuga simplex Wormsk

LI Ming,CHEN Xiaolin,YOU Ruilin

2000, 36(2):  186-192. 

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For the approach to the relations between the endangered situation and their sexual reproductive processes in Cimicifuga nanchuanensis, some comparative studies were made in C.simplex, a close and widespread species. The present paper deals with the microsporogenesis and the development of its male gametophytes and the anther wall. The anther possesses four microsparangia with its wall composed of four layers: epidermis, endothecium, middle layers and a 2-nucleate celled tapetum. Most of the antheral tapetum are in situ disintegrated. However, it is found that in about 12% of the locules, tapetual periplasmodia move into anther cavities forming so-called amoeboid tapetum when the microspore tetrads are released. The cytokinesis of microspore mother cells and wall formation is of simultaneous type and the tetrahedral microspore tetrads are formed. The mature pollen grains are 2-celled.

Construction, Expression, Purification and Characterization of GHRP-scu-PA-32K

JIAO Jianwei,XU Changfa

2000, 36(2):  203-208. 

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GHRP-scu-PA-32K cDNA was obtained by in vitro site-directed-mutagenesis to insert oligonucleotide sequence (GGTCATAGGCCT) encoding tetrapeptide Gly-His-Arg-Pro into scu-PA-32K gene at site just preceding the amino-terminal residue(Leu¹). The mutant cDNA was cloned in pCM-β-neo expression vector and co-transformed CHO-DHFR^- cells together with pCM-dhfr. The stable expression cell line was selected. Expression level of the line cultured in serum-free medium was 580IU/106 cell/24h. The product expressed was purified by Zn²⁺-Sepharose affinity chromatography. The apparent molecular weight of purified GHRP-scu-PA-32K was 33kD by SDS-PAGE and mass spectrometry and its specific activity was 150000IU/mg protein, according to fibrin plate determination. The affinity for fibrin of GHRP-scu-PA-32K was 4.5 times higher than that of scu-PA-32K.

Landscape Structure Study of Longhua Area in Shenzhen City during the Fast Urbanization Process
III Structure and Heterogeneity Analysis of Farmland

ZENG Hui,JIANG Ziying,YU Hong,XU Guoxiong

2000, 36(2):  221-230. 

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Remotely sensed mapping data and methods of landscape structure analysis were used to study the structure and spatial distribution characteristics of agricultural land patches during the fast urbanization process of Longhua area, Shenzhen City. Conclusions of the study were as follows: (1) The fast urbanization process made croplands decrease rapidly, but did not produce very significant impacts on the total area of the orchard; (2) Human landscape transformation during 1992 was very severely, and the cropland fragmentation could be divided into two different stages; (3) Random spatial distribution with microscopic heterogeneity of the two agricultural elements was maintained, but the evenness had a little bit decreasing. Compared with farmland, the spatial distribution of orchard expressed very different characteristics. Under continuous scale, distribution of orchard showed a regular pattern such as unevenness (small scale: side length of the box was shorter than 8 cells)-evenness (small and moderate scale: the length was among 8 and 50 cells)-unevenness (moderate scale: the length was among 50 and 200 cells)-evenness (moderate and large scale: the length was longer than 200 cells); (4) Human landscape transformation was the major impact factor of landscape changing, and a kind of moderate scale impact factor. The terrain was only a background impact factor; (5) During domestic landscape transforming in the working area, because the ecological maintenance value of two agricultural elements were not considered, their structural and functional equilibrium decreased gradually.

Thermoevolutional Characteristics of Hydrocarbon Generation from Barkinite, Late Permian Coals in Shuicheng, Guizhou Province

SUN Xuguang,WANG Guanyu,JIN Kuili

2000, 36(2):  209-213. 

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The Late Permian coal from southern China are unique in high content of barkinite and element hydrogen, as well as significant hydrocarbon-generation potential. In this paper, Rock-Eval, Py-Gc and MPV-3 were used to study the hydrocarbon-generation potential, the hydrocarbon composition, hydrocarbon-generation mechanism and thermoevlutional characteristics of barkinite in open system simulation. The results indicate that barkinite has higher hydrocarbon-generation potential, the values of hydrocarbon-generation potential (S₁+S₂) is 297.1mg/g; light hydrocarbon, wet gas and alkene are the main composition of hydrocarbon; the temperature range of hydrocarbon producting is at 400-450℃, the temperature range of hydrocarbon generation peak is at 420-450℃. These studies demonstrate that barkinite is one of the better macerals for petroleum generation. These studies will have greater values for exploring oil formation from coal from Late Permian coal in southern China.

The Research of Rock Poroelasticity

SHI Ge,YANG Dongquan,YANG Huizhu

2000, 36(2):  214-220. 

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The relation between poroelastic modulus and effective modulus of rock under dry and fluid-saturated conditions was discussed in this paper. The poroelastic modulus could describe pore effect on elastic behavior of porous media such as rocks. From this, Gassmann equation which was widely used in seismic exploration and petroleum logging was deduced. This equation was based on linear elasticity assumption and could be used only within low frequency. The accurate formula of computing effective pressure was pointed out also. The method was proposed to compute the rock modulus when one kind of pore fluid changed to another, and to identify the pore fluids by the use of poroelastic modulus.

Paleo-Environment Settings of the Dongfang Plaza Late-Paleolithic Site, Wangfujing, Beijing

MO Duowen,XU Haipeng,YANG Xiaoyan,ZHU Zhongli

2000, 36(2):  231-239. 

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The Dongfang Plaza cultural site, located in Wangfujing, Beijing, is the first found of late paleolithic relic in Beijing plain. The process of climate change during 26-14ka B.P. will be discussed in this paper, based on the analysis of the sedimentary facies characters and paleo-environment indexes such as clay mineral, chemical element and spore-pollen of the sediment samples collected from the middle-part of the stratigraphic profile (including upper and lower cultural layers) of the Dongfang Plaza cultural site. The landform on which ancient human activity relic located was interfluvial floodplain on Yongding River Alluvial Fan. The result of analysis shows that the climate of the period during which these strata were formed was mainly dry and cold, and the pollen of herbs is predominant. In the later part of this period the temperature and humidity increased a little, especially between the two cultural layers there was a short but clear section of wet and warm climate and a temperate forest steppe landscape existed. So it can be concluded that after the lower cultural stratum was formed, the flood became greater due to the wetter weather and bigger river discharge, and the ancient people were forced to move to other higher places. Later the climate began to dry and human activities appeared here again. In such a way two cultural strata were formed. Cultural sites of dry-cold late Late-Pleistocene have been found in Beijing, Inner Mongolia Autonomous Region, Heilongjiang province and other places, which indicates that the people of late Late-Pleistocene already had the ability to defend the dry-cold weather.

Recent Progresses in the Protein Engineering of Thrombolytic Agents

ZHANG Yanbin,XU Changfa

2000, 36(2):  286-294. 

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Arterial thrombosis is the underlying cause of a wide variety of cardiovascular diseases including myocardial infarction. All the currently used thrombolytic agents are plasminogen activators, which are very efficient in restoring the blood flow. They can convert plasminogen into plasmin and thus degrade fibrin. Despite the widespread use of established thrombolytic agents such as streptokinase, t-PA and u-PA, all these agents suffer from a number of inadequacies including resistance to reperfusion, occurrence of coronary reocclusion and bleeding complications. The quest continues for plasminogen activators with higher potency, specific thrombolytic activity, fibrin selectivity and longer half-life time. The recent progress in the protein engineering of plasminogen activators, including t-PA, u-PA, streptokinase, staphylokinase and other novel plasminogen activators, was presented in this article. We can conclude that the new generation of thrombolytic agents showed considerable potential with regard to improving the efficacy of thrombolytic therapy, and these progresses were based on the detailed understanding of the relationship between structure and function of thrombolytic agents. However, the risk of intracranial bleeding remains problematic and need more researches on the modification of thrombolytic agents.

Recent Progresses in the Protein Engineering of Thrombolytic Agents

ZHANG Yanbin,XU Changfa

2000, 36(2):  286-294. 

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Arterial thrombosis is the underlying cause of a wide variety of cardiovascular diseases including myocardial infarction. All the currently used thrombolytic agents are plasminogen activators, which are very efficient in restoring the blood flow. They can convert plasminogen into plasmin and thus degrade fibrin.Despite the widespread use of established thrombolytic agents such as streptokinase, t-PA and u-PA, all these agents suffer from a number of inadequacies including resistance to reperfusion, occurrence of coronary reocclusion and bleeding complications. The quest continues for plasminogen activators with higher potency, specific thrombolytic activity, fibrin selectivity and longer half-life time. The recent progress in the protein engineering of plasminogen activators, including t-PA, u-PA, streptokinase, staphylokinase and other novel plasminogen activators, was presented in this article.We can conclude that the new generation of thrombolytic agents showed considerable potential with regard to improving the efficacy of thrombolytic therapy, and these progresses were based on the detailed understanding of the relationship between structure and function of thrombolytic agents.However, the risk of intracranial bleeding remains problematic and need more researches on the modification of thrombolytic agents.

Activity Research for the Dalinghe Hidden Fault

WAN Bo,GAO Changbo,WU Ge,WU Mingda,HOU Jianjun

2000, 36(2):  240-245. 

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The Dalinghe hidden fault, as an example in this paper, is just located under the Dalinghe river, so the activity of this hidden fault can be defined by comparing Quaternary layers and terraces beside the river (hidden fault). Based on spore analysis of 17 layers and several ¹⁴C chronological data, the T₁ terrace of Dalinghe river section from Dayushupu to Linghai city is formed between Q₃ and Q₄; T₂ formed in Q₃; T₃ formed between Q₂ and Q₃¹. The same grade terrace height beside the river is surveyed at almost the same level, proved that the hidden fault under the river never moved after Q₃ when T₃ formed. The average crust up rate in this area from Q₃ till now is 0.20-0.28mm/a.

The Effect of Lithology and Climate on Major Ion Chemistry of the Yangtze River System

XIA Xinghui,ZHANG Litian,CHEN Jingsheng

2000, 36(2):  246-252. 

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Based on the analyses of water quality monitoring data of over 120 hydrometric stations in the Yangtze River System during the last 30 years, the effect of lithology and climate (rainfall) on major ion chemistry of the river water has been studied by means of statistical approaches such as principal component analysis (PCA) and correlation analysis.

Synthesis, Characterization and Magnetism of Cyano-bridged Cu₂^IIFe^II and Ni₆^IIFe^II Complexes

SUN Xianru,CHEN Zhida,BIAN Jiang,WANG Shixi,AN Jin,XU Guangxian

2000, 36(2):  253-258. 

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Two cyano-bridged complexes {[Cu(L)₂]₂[Fe(CN)₆]}_∞(L=NH₂CH₂CH₂CH₂OH(ampt) (1), NH₂CH₂CH(CH₃)OH(amtt)(2)) and cyanobridged Ni₆^IIFe^II cluster [Fe(CN-Ni(tetren))₆](ClO₄)₈(tetren)(3) were synthesized, and have been characterized by elemental analysis and IR spectrum. The experimental results show that the oxide state of Fe is changed from Fe^III to Fe^II after the synthesis reaction. The magnetization versus T and magnetic susceptibility of (2) were measured and showed that the interaction between Cu^II ions separated by diamagnetic low spin Fe^II is weak antiferromagnetic.

Identification and Expression of the Product from Engineering Bacterium Containing Recombinant Human Interleukin-18(IL-18) Gene

DONG Yuqing,LI Ronggui,DONG Chunxia,ZHAO Jindong,WU Guangyao

2000, 36(2):  271-274. 

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The cDNA of human Interleukin-18 was obtained by RT-PCR from the mRNA of human peripheral blood cells. The DNA sequencing was performed and the DNA was cloned into expression vector pBV220. The overexpression was performed in E.coli HB101. The expressive product was purified through Sephadex G100 and identified by N-terminal amino acid sequencing and Western blot.

Screening of Combinatorial Libraries and Its Application in Drug Discovery

WU Zengru,LI Youyong,XU Xiaojie

2000, 36(2):  275-285. 

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How to determine the structures of active compounds in combinatorial libraries quickly and efficiently is a challenge that combinatorial chemists must face since the beginning of combinatorial chemistry. It is the foundation of high throughput screening. The article reviews the progress that combinatorial chemists have made in recent years. The first part discusses that the structures of active compounds can be determined through permutation and combination of the libraries; The second part is about the different techniques of encoded libraries; In the third part, some methods of analysis the active compounds directly are discussed; At last the efforts we have made in the field of combinatorial chemistry are introduced.

Study of NMR and FT-IR Spectra of Bis(substituted Cyclopentadienyl) Dichlorides of Titanium and Zirconium

ZHANG Jianbo,XU Zhenhua,CHEN Shoushan

2000, 36(2):  259-262. 

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The NMR and FT-IR Spectra of ten (η C₆H₉CRR'-Cp)₂MCl₂(M=Ti, Zr) were measured. The constructions of the compounds were identified by ¹H NMR and ¹³C NMR spectra. All complexes are bonded by M-Cp through center of Cp. The influence of the center metal atoms and the substituents upon the NMR and FT-IR spectra were discussed.

Cloning and Primarily Detecting the Function of the Promoter Region of NFL Gene in Tobacco

LIU Fuquan,ZHANG Yanwen,XU Zhihong

2000, 36(2):  263-267. 

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The promoter region of NFL1 gene in tobacco was cloned by a novel PCR method, fused with the report gene GUS and transformed tobacco plants. The expression of GUS in the shoot apex of transgenic seedlings was detected.

Crystallization and Preliminary X-ray Studies of Metallothionein-II from Rabbit Liver

AN Yu,LI Genpei,RU Binggen

2000, 36(2):  268-270. 

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Metallothionein-II, 6.8kDa, purified from the liver of rabbit induced by cadmium, has been crystallized in the space group P6₃22 using hanging drop vapor diffusion method, with unit cell parameters a=b=11.34nm, c=14.66nm. There are 8 molecules per asymmetric unit and a solvent content of 58%.

Screening of Combinatorial Libraries and Its Application in Drug Discovery

WU Zengru,LI Youyong,XU Xiaojie

2000, 36(2):  275-285. 

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How to determine the structures of active compounds in combinatorial libraries quickly and efficiently is a challenge that combinatorial chemists must face since the beginning of combinatorial chemistry. It is the foundation of high throughput screening. The article reviews the progress that combinatorial chemists have made in recent years. The first part discusses that the structures of active compounds can be determined through permutation and combination of the libraries; The second part is about the different techniques of encoded libraries; In the third part, some methods of analysis the active compounds directly are discussed; At last the efforts we have made in the field of combinatorial chemistry are introduced.