Abstract:In recent years, the isthmo-optic nucleus (ION) in the avian centrifugal system became a new model to study the apoptosis and the development of central nervous system (CNS). During its development, together with the formation, folding and laminating of the nucleus, there exist some temporal pathways linked to the nucleus. More than half of its neurons befall apoptosis in this period. Researches show that proper efferent and afferent projections are essential to the survival of neurons. The molecular mechanism indicates that apoptosis is related to a series of neurotrophins and their receptors. Apoptosis plays an important role in the development of CNS.

Abstract:Recently lot of research indicated that pluripotent neural stem cells with self-renewal and multilineage potential are present in the adult mammalian forebrain. The forebrain tissue containing persistent neural stem cells is referred to as “brain marrow”, since it has much in common with hematopoietic bone marrow. Neuropoiesis are very important field in neuroscience. A thorough investigation of neuropoiesis in the brain marrow should facilitate utilizing of migrating neurons in the adult human forebrain and transplanting human ES cells to treat neurodegeneraton diseases.

Abstract:The technique of pronuclear microinjcetion is the most reliable and popular method for transgenic livestock production to date. However, pronuclear microinjection has been proven to be inefficient in producing transgenics and in inserting a piece of DNA into a specific site in the host genome. In the past 20 years, some new approaches have been used. These include sperm-mediated DNA transfer, retroviral mediated DNA transfer into oocytes, somatic cell carrying exogenous genes nuclear transfer, the use of embryonic stem(ES) cell. But these ways can not solve existing problems thoroughly. In recent papers, the existing techniques have been improved and significant progress has been made in developing transgenic techniques.

Abstract:D-glucose isomerase(GI) can isomerize D-glucose and D-xylose into D-fructose and D-xylulose，respectively. It is a crucial enzyme in the production of high fructose corn syrup on industrial scale. In the hydride shift mechanism proposed for GI，the main features are ring opening of the substrate，isomerization of GI via a hydtride shift from C₂ to C₁，and ring closure of the product. GI gene has been cloned，sequenced and overexpressed in the homologous and heterogous hosts. GI whose properties have been improved by protein engineering may be one of the most important industrial enzymes of the future.

Abstract:Vascular endothelial growth factor(VEGF) induces angiogenesis as well as permeability of blood vessels,and plays a central role in the regulation of vasculogenesis.Five VEGF isoforms (VEGF_121～206) are produced from a single VEGF gene by alternative splicing of the VEGF mRNA.The regulation of angiogenesis induced by VEGF and how to inhibit its effects arouse great interests.The function of VEGF in angiogenesis regulation was reviewed in several aspects.

Abstract:Plant homeotic genes and homeobox genes are two types of the important genes encoding transcription factors involved in plant development. Research during recent ten years indicates that there are differences between their structures and functions. It is very important to study their structures and functions and reveal plant developmental mechanisms.

Abstract:By mediating and regulating of apoptosis in macrophages themselves and other cells as well, macrophages carry out their function in immunology and immunological regulation. The macrophage apoptosis inducing factors were of biologic, chemical, pathologic. Some distinguished features characterized macrophage apoptosis and its regulation. Interestingly, to fit in with need of the body, macrophages may mediate or inhibit apoptosis in themselves, favour or inhibit apoptosis of other cells, inhibit apoptosis in themselves and favoured apoptosis of others, which may be the base for macrophages play a role in immunological regulation, especially in tumor immunology.

Abstract:FEN-1 is a kind of structure-specsific nuclease, which could recognize specific DNA bifurcated structure and cleave single-strand DNA with free 5′ end. During DNA replication, FEN-1 5′→3′ exonuclease removes the last nucleoside of RNA primer attached to an Okazaki fragment. In DNA repair, FEN-1 5′→3′ endonuclease is involved in the process of removing the damaged nucleotide. FEN-1 gene contains two conserved regions and one PCNA binding region.

Abstract:A conserved family of aspartate-specific cysteinyl protease(Caspase) has been identified as mediators and excutors of programmed cell death (PCD) in mammalian cell. Proapoptotic signals culminate in activation of different initiator caspase which，in turn，activate effector caspase through enzyme cascade pathways. Active effectors cleave a set of substrates and result in cellular disassembly. Caspase family is the critical elements in PCD. They regulate cell death or survival by interaction with many proteins(activators or inhibitors).

Abstract:Pichia pastoris has been utilized widely as a heterologous gene expression system recently. There are several advantages such as high productivity, stable inherity, secretable product, and mature fermentation techniques. The advances of vector types, vector elements(including promoter, selection marker and signal sequence), host strains, and the strategy for increasing integration copy number were reviewed.

Abstract:PSⅠ, PSⅡand light-harvesting complexes (LHC) in oxygen evolving photosynthetic organisms were reviewed. These organisms include cyanobacteria, red algae, brown algae, diatoms,chrysophytes,dinophytes, xanthophytes,crypophytes,green algae and green plants.The diversity of pigment-protein complexes that fuel the conversion of radiant energy to chemical bond energy was highlighted, and the evolutionary relationships among the LHC structural polypeptides and the characteristics of the fluorescence emission of PSⅠat 77 K was discussed.

Abstract:Calcineurin(CaN), a Ca²⁺/calmodulin-dependent protein serine/threonine phosphatase, broadly distrbutes in various mammalian cells and involve into regulation of cellular function. It was known that CaN play a central role in T cell activation and is essential to transmitter release and synapic plasty. It is reported recently that CaN is likely a link of Ca²⁺ signal with cardiac hypertrophy. The advances in the molecular structure, enzymatic propertied, distribution and biological function of CaN were summrized.

Abstract:With the technological developments of cryoelectron microscope, X-ray diffraction and the growing data available on various components of ribosome, some marvelously intricate structural models of the Escherichia coli 70S ribosome have been reconstructed. The picture of the ribosomal model are detailed, including the placement of the mRNA, the arrangement of the A-site and P-site tRNAs and the peptidytransferase within the interface gap as well as the path of nascent polypeptide chain, which results in a better understanding of the structure and function of ribosome as well as the translational process.

Abstract:Genome annotation using bioinformatics tools becomes one of the most active research fields in the postgenome era. The new development in this area was reviewed. Various levels of genome annotation are discussed, while predicting protein function in genome scale is well discussed.

Abstract:Prolyl endopeptidase(PEP) ［EC 3.4.21.26］ is a new class of serine protease, which is capable of hydrolyzing a substrate on the carboxyl site of proline residue located internally in a peptide. It can hydrolyse several peptide hormones and neuotransmitters, so abnormal raise and decrease of PEP activity would result in diseases related to memory and cognition. Specific inhibitor for example JTP-4819 shows a good pharmaceutical effect in reversing scopolamine-induced amnesia in rats. The analysis of crystral structure of PEP from porcine muscle has greatly promoted the research on PEP.

Abstract:Short-term synaptic plasticity is an important expressing form of synaptic plasticity and plays an essential role in the realization of the neural system’s normal functions. Short-term synaptic plasticity can make neural information transmission reliable, adjust the balance between excitation and inhibition of cortex, form the spatio-temporal properties of neurons’ activity, form and adjust the coherent oscillations in cortico-thalamic networks. Short-term synaptic plasticity may participate in some high-level brain functions, such as attention, priming, sleep rhythm, learning and memory.

Abstract:To increase the stability and bioavailability of antisense oligodeoxynucleotides, and to avoid being destroyed by the lysosomes, pH-sensitive prelip-osomes were preparaed by using of dehydration-rehydration, ultrasonication, extrusion, lyophilization methods. The effects of different variables on the preparation of pH-sensitive preliposomes were studied. The optimized preparation conditions of pH-sensitive preliposomes were acquired through orthogonal test. The experiment revealed that the pH-sensitive liposome was regular in its morphology with a mean diameter of 22.7 nm. The mean entrapment efficiency of antisense oligodeoxynucleotides of three batches was 68.3%. The release properties could be expressed by the following equation: Q=1.8382－2.5186×10^－2T(r=0.9913).Based upon the various assays used to measure the entrapment efficiency and diameter of pH-sensitive liposomes, it was concluded that the entrapment of antisense oligodeoxynucleotides by pH-sensitive preliposomes was effective.

Abstract:During the process of sodium butyrate (NaBu)-induced apoptosis in 2BS cells, p21^WAF1 gene expression decreased obviously and gradually,bcl-2 gene expression did not decrease until apoptosis happened, c-myc and c-fos genes expression enhanced, while p53 and HER-2 genes expression had no obvious change. To analyze the change of p21^WAF1 promoter activity, 2BS-WP cells were used, which were stabely transfected with pDOR-WPs that containing different length of p21^WAF1 promoter followed by green fluorescence protein (GFP) report gene. The results showed that the level of GFP expression decreased during NaBu induction, and the main regulating region was 0～－800 bp at upstream of TATA box of p21^WAF1 promoter. It is suggested that down-regulation of p21^WAF1 promoter activity was involved in the process of NaBu-induced apoptosis in 2BS cells, which was possibly p53-independent.

Abstract:By the investigation on different regions of the yolk of fertilized eggs, it was discovered that DNA and RNA were widely distributed in the yolk. DNA and RNA in the yolk were quantified by the method of fluorescence spectrophotometry with the reagents Hooechst33258 and RiboGreen respectively. The results indicated that DNA was uniformly distributed in the whole yolk(about 65.7 μg/L) and it was most probably that the DNA was contained in the yolk granules; while the distribution of RNA in the yolk showed another pattern: the amount of RNA in the superficial yolk beneath blastoderm was about 0.43 mg/L, which was much less than that of lateral yolk(2.87 mg/L) and vegetative pole(3.63 mg/L). The significance of the existance of yolk DNA and the unique distribution of yolk RNA remains to be elaborated.

Abstract:An adeno-associated viral vector containing erythroid enhancer(HS2), genomic β-globin gene and neomycin resistance gene was constructed, then packaged into recombinant AAV virions. Southern analyses showed that recombinant provirus genome steadily integrated into erythroid cells via virus transduction. It was suggested that an adeno-associated virus could mediate stable integration of genomic gene of interest in mammalian cells.

Abstract:A NADPH-dependent 3-DG metabolizing enzyme was isolated and purified to electrophoretic homogeneity from Bacillus sp.2 by combined consecutive treatment consisting of ammonium sulfate fractionation, Q Sepharose FF, Sephadex G-100(Ⅰ), Hydroxyapatite and Sephadex G-100(Ⅱ) column chromatographies. The specific activity of purified 3-DG metabolizing enzyme was 63.75 U/mg. The molecular weight of the enzyme was about 32 ku. 2-Oxoaldehyde compounds were found to be specifically good substrate for this reductase. The optimum pH of the enzyme activity was 6.2. The enzyme was stable in the pH range from 5 to 8 and in the temperature range from 25℃ to 30℃. The K_m for 3-DG was 2.3 mmol/L. Suitable amount of EDTA、β-mercaptoethanol and dithiothreitol enhanced the enzyme activity, but the activity of the enzyme was partially lost by adding iodoacetic acid or N-ethylmaleimide.

Abstract:To investigate the mechanism involved in the apoptosis of human monocyte cell line U937 induced by oxidized low density lipoprotein (ox-LDL).The increase of the degree of cell apoptosis was concentration dependent. The apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL),flow cytometry and DNA fragment analysis. The protein contents of c-fos，c-jun and c-myc were analyzed by immunochemical staining. The expression of c-fos，c-jun and c-myc mRNA were quantified by reverse transcription-polymerase chain reaction (RT-PCR).The results show that ox-LDL could upregulate the gene expression of c-fos，c-jun and c-myc, which induced the apoptosis of U937 cells.

Abstract:Protein kinase CK2 is a heterotetramer ser/thr protein kinase composed by two catalytic subunits (α or α′) and two regulatory subunits (β). The recombinant plasmid containing human CK2β subunit cDNA was transformed into E.coli BL21(DE3) and expressed induced by IPTG. Most of the expressed CK2β proteins were insoluble. From 6 L(about 10.4 g) bacteria, 20 mg soluble protein was extracted from the insoluble pellet and purified by a single step P11 phosphocellulose chromatography, the yield of purified CK2β subunit was 6.8 mg. SDS-PAGE analysis of the purified protein showed only one band with molecular mass of 26 ku. Western blot analysis confirmed that the expressed product was human CK2β subunit. Addtion of the CK2β subunit to CK2α subunit led to maximum stimulation at a 1∶1 molar ratio of both subunits. These results demonstrated strongly that the cloned, expressed and purified recombinant protein was human CK2β subunit. The large amount of purified recombinant CK2β protein lays solid basis for further study directly the characteristics of the enzyme and the relationship of structure and function between CK2β subunit and its interacted proteins.

Abstract:PCR based method for differential display of eukaryotic mRNA has been designed to isolate differential expressed genes in various cell types or under different growing conditions. A modified method for mRNA differential display originally developed by Sokolov was employed and optimized here. This procedure, based on the application of Ready-To-Go RT-PCR beads and Ready-To-Go RAPD beads, minimized pipetting steps, decreased the potential for pipetting errors, reduced the risk of contaminating and ensured greater reproducibility between reactions. Distinct cDNA bands can be observed by silver-staining 6% sequencing urea gel and easily excised and recovered for further use in cloning. The stage-specific genes in the developing human embryos were analyzed with six sets of arbitrary primers using this modified DDRT-PCR from 3-, 4- and 5-week-old human embryos. About 14 bands containing differential fragments were obtained from sliver-straining 6% polyacrylamide gel, six of which were proved to be developmental related genes by RNA rev-Northern hybridization using ［α-³²P］dCTP labeled first strand cDNA as probes.

Abstract:A fast staining-destaining method for SDS-PAGE which favours the recovery of proteins was developed:brief staining by Coomassie Brillant Blue R-250 followed by destaining with 250 mmol/L KCl.The SDS-PAGE stained-destained by this method has almost the same clearance compared with the one stained-destained by the usual method,and the recovery of protein can be enhanced more than three times by this method.

Abstract:Polypyrrole doping anions was obtatined with methods of chemical polymerization and electrochemical polymerization.Nervous tissue was cocultured with polypyrrole and its biocompatibility was observed with an inverted optical microscope and a scanning electron microcope.It is concluded that polypyrrole has good biocompatibility with nervous tissue.

Abstract:PCR products amplified with Taq and Taq+Pwo DNA polymerase were reamplified by using UT-PCR and then cloned and sequenced. The termini of the original PCR products were analyzed. In the group of Taq amplification, the termini were chiefly sticky ones with 3′-protruding A(67.3%);the next majority of termini were blunt ones(26.9%).In the other group, the proportion of blunt-ends was almost the same as that in the group of Taq amplification(26.1%). However, the 3′-protruding termini decreased(17.4%),instead of additional 3′-recessive ones(－1,－2,－3;57.0%). The results reveal that the ends of PCR products are complicated.

Abstract:To investigate the differentiation of pre-preparation method to determine CD4 on T lymphocyte in blood, CD4 expression on T lymphocyte of monkey was determined by the flow cytometrey as a quantitative analysis with three different pre-preparation methods. These results showed that (b) method (whole blood was labeled by FITC-CD4 McAb. Then red blood cells were dissolved by erythrocytolysis) was better (a) method (red blood cells were dissolved by erythrocytolysis in whole blood. Then it was labeled by FITC-CD4 McAb). CD4 results with (b) method were equal to (c) method (traditional method of lymphocyte separated). But the volume of sample by (b) method only was 1/5 of (c) method. The pre-preparation of (b) method also was sampler than (c) method.Morphologic study by laser scanning confocal microscopy showed (b) method was similar to (c) method. Both of them had a clear fluorescent labeled on the cell membrane but not in the cell.

Abstract:Low-level chemiluminescence exist in the most biological systems. The chemiluminescence is relate with physiology response of organism. The physiology variety of organism is caused if the organisms living in the water and air were endangered by various contamination factors, consequently change of the chemiluminescence is produced. Using the phenomenon, Some organism can be used as a biological detector, environment grade of water and air can be examined by ultra-weak chemiluminescence analytical technology.