Abstract:The cell cycle is a successive process between two mitosis divisions. During these courses, the cellular genetic materials, for instance DNA, are equally assigned to two daughter cells after replication. Each cell is endowed with its own share of the parental chromosomes. The schedule of events in cell cycle happens in the presence of regularity, precision and perfect temporal and spatial restriction. CDKs, cyclins and cdc are essentially key regulators of three families in control of cell cycle. The discovery of these regulators has important values on both basic research and applied biotechnology.

Abstract:β-amyloid(Aβ) is a predominant component of senile plaques, one central pathological hallmark of Alzheimer's disease. Immunization with Aβ causes a marked reduction of Aβ burden and its associated pathologies in brain. A clearance of the preexisting plaques has been observed in transgenic mice, with an inhibition of lesion of cognitive functions. No signs of an immune-mediated response could be detected to endogenous Aβ peptide of mice. The chronic inflammatory response resulted from the immunization itself has not obviously impaired functions of central nerve system.

Abstract:Antisense-oligonucleotide can be employed as gene expression inhibibor and potential therapeutic agents. However,many classes of oligonucleotide are polyanion and cannot penetrate cell membrane. It is known that several peptides,including fusion peptide and signal peptide, have the properties of transmembrane and/or cell nucleus localization.The synthesis and biological activity of antisense oligonucleotide-peptide conjugates are discussed.

Abstract:There are hundreds and thousands kinds of biomacromolecules within cells, proteins, nucleic acids, polysaccharides and so on, and the total concentration of those macromolecules could be high up to 80～200 g/L. In general, cellular interiors are 20%～30% volume-occupied physically by macromolecules, and such an intracellular environment has been termed as “macromolecular crowding” or “the excluded volume effect” more precisely. The biophysical theory has predicted significant effects of macromolecular crowding on biochemical reactions thermodynamically and kinetically, however, the important aspect of the intracellular environment is largely neglected. It has been strongly suggested that addition of crowding agents at biologically relevant concentrations to working system should become a routine variable just like pH and ionic strength to make the study under more physiologically relevant conditions.

Abstract:Orphan receptor TR3/nur77, a member of nuclear receptor superfamily, is a product of immediate-early gene which is expressed rapidly after induced by several proliferation factors such as EGF, FGF, NGF, PDGF or phorbol ester, and different apoptosis inducers. It is similar in structure with the members of steroid/retinoid receptor superfamily that all have a DNA-binding region of two zinc-finger and a ligand-binding region. As a transcription factor its complex functions are involved in proliferation, differentiation and apoptosis of cells. Recently there is an important development about the TR3 receptor in the mechanism of inducing apoptosis. After treated by different apoptosis inducers TR3 expressed increasingly, and it is found surprisingly that TR3 translocates from the nucleus to mitochondria to induce cytochrome c release and apoptosis. That is, mitochondria targeting of TR3 but not its DNA binding and transactivation is essential for its proapoptotic effect. This is a new model may be a very significant pathway in the regulation of cell signal transduction especially in the apoptosis.

Abstract:The genomic-scale EST and genome sequencing, and cDNA microarray analyses that are now under way promise to rapidly isolate and identify all candidate genes essential for tolerance of osmotic potential, desiccation or temperature stresses. The large datasets generated by these efforts will provide basis for functional analysis with the use of tagged mutant collections, complementation and overexpression tests accompanied by microarray analyses. By using yeast two-hybrid technologies, the specific protein-protein interactions will be well-understood. When the functions of all genes that participate in stress adaptation or tolerance reactions are determined, an integrated understanding of the biochemical and physiological basis of stress responses in plants will be obtained, and then, it will be therefore possible to rationally manipulate and optimize tolerance traits for improved crop productivity.

Abstract:The transcription factor “decoy” strategy had been reported that applying double-strands oligodeoxynucleotides(ODNs) transfect target cells and compete with the sequence of endogenous cis-element for binding to transcription factor, thus leading to prevention of the endogenous gene expression. “Decoy” strategy was not only a novel strategy for gene therapy but also a powerful tool for the study endogenous gene regulation.

Abstract:In order to arrive at a complete description and understanding of brain function and anatomy, the neuroimaging techniques are required to provide both high temporal resolution and high spatial resolution. Multimodal imaging that combining hemodynamic and electrophysiological information (i.e., fMRI/PET and EEG/MEG), holds promise for imaging patterns of human brain activity in both space and time. Multimodal imaging has succeeded in revealing the spatiotemporal pattern of brain activity underlying selective attention, visual perception of kinetic form, voluntary movement and semantic processing. However, there are still much work to do to improve the accuracy and the spatiotemporal resolution of multimodal imaging to elucidate the neural mechanisms of human cognition.

Abstract:lats gene (large tumor suppressor gene), first identified in Drosophila melanogaster, has its homologues in mouse and human. The function of lats is highly conserved from Drosophila to human. Its function includes: as a tumor suppressor gene, lats mutation will result in a tumor phenotype; phosphorylated Lats can bind to Cdc2 and regulate cell cycle; Lats may also be involved in size control mechanism via cell-cell communication. The study of lats function from Drosophila to human can provide a method to study gene functions in mammals using Drosophila as a model organism.

Abstract:The progress of molecular biology attributes the complexity of life to the interaction of mass genes ,and thus the traditional descriptive method in biology and analysis by disassembling faces the cruel challenge. The application of DNA chip and molecule array enables scientists to monitor the massively parallel expression of genes,which lead to emphasis of gene networks as a systematic,quantitative method in the research of life science.Based on the crossing of subjects on molecule biology, nonlinear maths and informatics,the object of gene networks is to construct the dynamics model which has nonlinear traits such as robustness,hierarchy and so on.The massive data of gene expression combined with fit algorithm can help scientists establish the topology of the interactions,by which the system behavior can be simulated. Contrariwise,the model established can direct the further experiments.To study gene networks the computer and Internet resource are very important.The study of gene networks will play an important role in the post-genome research.

Abstract:Protein array/chip technique is a significantly-advancing technology on molecule biology.There is wide latent application perspectives in function proteome study.The present developing printing protein microarray, molecular scanning technique and biosensor chip mass spectrometry will apply to the aspects to detection medicine targets, diagnosis diseases,identification proteins,and/or interaction between proteins, etc.Because analyses on these chips have many advantages such as quick detection, high efficiency,little sample consumed and low cost,they will become novel study tools in the field of life science and medicine.

Abstract:DNA microarray for gene expression profiles is a high throughput technique. It allows expression monitoring of tens of thousands genes in parallel. It has become an increasingly popular tool to investigate the function of genes. Here the principle and applications of this technique were reviewed. The problems it faced with and the resolve strategies are also mentioned.

Abstract:The nearly close of sequencing stages of the various genome projects transforms structural biology into structural genomics. Structural genomics is the systematic determination of all genome products' structures. It uses high-throughput selection, expression, purification, structure determination and computational analysis to provide an experimental structure or a good model for every protein in all completed genomes, that will accelerate scientific study in all areas of biological science. The developments of bioinformatics, gene engineering and structure determination techniques provide the guarantee for structural genomics. Recent developments in the technology of nuclear magnetic resonance make it as a key method of high-throughput structural analysis in structural genomics.

Abstract:The loading of peptide into ultrafine host vesicles or in the nanometer size range is an important technique for the optimization of controlled peptide antigens delivery.There are closed relationship between the nanoparticle and the organism,the size range of DNA-protein complex is 15～20 nanometer.Several virus particles are also nanoparticle.For the nanoparticle adjuvants,the carrier effects and side effects and foreign body irritation will be avoid.Furthermore, the actual state of the nanoparticle materials is shown with 4 practical application examples,that is: a targeted cellular uptake by macrophage and dendritic cells; the strong immunity stimulation of nanocapsules, as new adjuvants, when loaded with viral or other antigens; the better blood-brain barrier transfer of a biochemistry drug when covalently bound to special liposomes; and the use of minivesicles for controlled site-specific anticancer drug release (tumor targeting).The nanoparticl materials were manufactured by physics and chemistry methods.Almost all biochemistry drug and DNA drugs, particularly, adjuvants for the epitope peptide vaccine were loaded with nanoparticle carriers for the good stability and body-friendly and biodegradable excipients.

Abstract:Since the first report of successful spermatogonial transplantation issued by Brinster's laboratory in 1994, some important new findings has been revealed by a series of recently studies relating to spermatogonial transplantation. It has been found that germ cell can be transferred not only from one animal to another, but also from one species to another，which is a big technological breakthrough in the male reproductive researches, especially for the study of Sertoli-germ cell interaction. In essence, Brinster's laboratory has pioneered another transgenic technology that is capable of modifying the entire genome of the animal.

Abstract:In order to study the neutrophil gelatinase-assiciated lipocalin(NGAL)gene expression character in the progress of malignant transformation of human immortalized esophageal epithelial cell, differentially expressed NGAL gene was identified by using cDNA microarray in the human immortalized esophageal epithelial cell line(SHEE) and malignant transformed esophageal cancer cell line(SHEEC). NGAL expression profile was further confirmed by Northern blot and RT-PCR. A cDNA encoding NGAL from SHEEC was amplified by PCR and sequenced. Alignment was analyzed by NCBI database. The results indicated that NGAL gene was overexpressed in the SHEEC. The coding region cDNA of NGAL from SHEEC was cloned and identified. Alignment of its deduced amino acid sequence compared to the mouse 24p3 protein, the rat neu-related lipocalin(NRL), the human NGAL from neutrophil and ovarian cancer demonstrated a very high degree of conservation. It can be concluded that NGAL overexpression possibly played an important role in the progress of malignant transformation of human immortalized esophageal epithelial cell. NGAL may be a new oncogene or promoter-tumor gene.

Abstract:The optical storage characteristic of the D96N genetic mutant bacteriorhodopsin film is experimentally studied, where the optical image is recorded on the BR film with 670 nm laser, the readout of the image is carried out by a weak 560 nm light, and the erase of the image is completed with the illumination of 488 nm laser. The lifetime of the M state is measured to be extended to 3 minute at room temperature, which is five-order magnification longer than that of the wild type bacteriorhodopsin in the solution.

Abstract:Single strand conformation polymorphism (SSCP) has been widely used for mutation detecting. But the theory about it is not strong enough to guide application. Recently, many computer programs have been designed for predicting the folding of single strand DNA or RNA. Second structures of three DNA segments with RNAstructure 3.2 have been predicted and have been compared with their results of SSCP. The comparison shows there is closely relation between them. The kinds of prediction can guide operator to select primers for SSCP analysis and accurately estimate mutated ratio.

Abstract:HWAP-Ⅰ is a peptide neurotoxin purified from the crude venom of the Chinese bird spider Selenocosmia humena, which has analgesic activity. According to the RT-PCR result, an artificial gene encoding HWAP-Ⅰwas chemically synthesized. The gene was then inserted into pPIC9K,a secretory expression vector for Pichia pastoris.The constructed pPIC9K-HWAP-Ⅰ was transformed into his⁴ mutant yeast GS115 and a Mut^s His⁺ cell line was screened.The recombinant HWAP-Ⅰwas secreted into the culture supernatant induced by 0.5% methanol.Tricine SDS-PAGE analysis indicated efficient expression and secretion of HWAP-Ⅰ.The molecular mass of product was about 4 ku. Yield of product was estimated to be 80 mg/L. Purified protein was assayed for biological activity and the results demonstrated that HWAP-Ⅰ had 60% analgesic activity compared to native HWAP-Ⅰ.

Abstract:Three different forms of PSⅠ complexes were isolated from a siphonous marine green alga, Codium fragile, by Triton X-100 sucrose gradient centrifugation. Zone Ⅲ had a Chl a/b＞20, and designated as PSⅠ core complex CCⅠ because it created only CPⅠ band in mild PAGE. Zone Ⅳ and Ⅴ had absorption at 436 and 674 nm, 467 and 650 nm, and 540 nm, suggesting the presence of Chl a, Chl b, siphonaxanthin and siphonein, Chl a/b were 3.23 and 2.4, respectively. Both CPⅠ and CPⅠa bands were observed when they were subjected to mild PAGE. Therefore, Zone Ⅳ and Ⅴ were different forms of PSⅠ complexes that consisted of CCⅠ and different amount of light-harvesting complex LHCⅠ. Zone Ⅲ contained only 66 and 56 ku peptides in SDS-PAGE, while Zone Ⅳ and Ⅴ had 4 different LHCⅠ peptides of 25，26，26.2 and 27.5 ku in addition to 66, 56 ku peptides. Fluorescence emission spectra showed that efficient energy transfer were kept among pigments in isolated PSⅠ complexes. Excitation energy absorbed by Chl b, siphonaxanthin and siphonein can be transferred to Chl a.

Abstract:A novel plasminogen activator containing low molecular single-chain urokinase (scuPA-32k) and fibrin β chain polypeptide (Fβ 15～42) was designed and constructed. ScuPA-32k cDNA was obtained by polymerase chain reaction (PCR) from pro-urokinase gene; while Fβ (15～42) cDNA was generated by joining synthesized oligonucleotide fragments together. Through suitable linker and approximately restriction site, scuPA-32k and Fβ (15～42) cDNA were ligated together. The fusion protein was expressed by IPTG induced in E.coli. After denaturation and renaturation, the aim protein was purified to homogeneity by Zn²⁺ chelating chromatography and Sephacryl S200 chromatography. The apparent molecular mass was 35 ku shown by SDS-PAGE analysis. The special activity was 87 000 U/mg detected by fibrin plate determination. The enzyme had similar kinetic parameters to that of natural uPA-32k when was assayed with the chromognic substrate S2444. However Fβ(15～42)/scuPA-32k had higher fibrin affinity than that of natural scuPA-32k and had antifibrin polymerization. These results showed that the fusion protein had good respects.

Abstract:A grass carp （Ctenopharyngodon idellus）cell culture ZC7901 was used as an in vitro model system to study the effect of heavy metal ions on fish cells. The results showed that the cell culture chemically stressed by six heavy metal ions, Cd²⁺，Cr⁶⁺，Hg²⁺，Cu²⁺，As⁵⁺ and Pb²⁺, exhibited cell death accompanied by various morphological alterations such as nuclear condensation, chromatin abutted sharply against nuclear membrane and developed into distinct apoptotic-like bodies, which were characteristic features seen in apoptosis. By agarose gel electrophoresis, DNA ladder were clearly visualized, which indicated the specific degradation of DNA. Also terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) analysis of the cells demonstrated DNA fragmentation localized in the dying cells as well as in the additional apoptotic-like bodies. All the results indicated that six heavy metal ions, Cd²⁺，Cr⁶⁺，Hg²⁺，Cu^{2+，As5+ and Pb2+, were all able to induce apoptosis in fish cells.}

Abstract:In order to study effect of extracellular Ca²⁺ and ionophore A23187 on erythrocyte deformation, the new ektacytometry is used. After erythrocytes were treated with different concentrations of intercellular Ca²⁺ and ionophore A23187 respectively, the orientation index (DI)_or and small deformation index (DI)_d of the erythrocytes were measured. It is suggested that the effect of ionophore A23187 on the rheology of erythrocytes is more notable than that of intercellular Ca²⁺. Furthermore, the (DI)_or,max and (DI)_d,max decrease as the concentration of intercellular Ca²⁺ or ionophore A23187 is increased. The deformability of erythrocytes was obviously reduced after treated with ionophore A23187.

Abstract:The pollution of heavy metals may affect the human health, in order to study the affect of heavy metals to the brain, the normal alive rabbits and the rabbits which were injected with PbCl₂ for 2 weeks were autopsied, the brain proteins were extracted and separated by gel filtration, and further analyzed by 2-D PAGE, More than one hundred water-soluble proteins of rabbit brain were visualized by Coomassie Brilliant Blue staining. The apparent molecular masses of most water-soluble proteins from rabbit brain were about 6～30 ku, pI was about 4～9. After matched with normal alive rabbit brain protein spots separated by 2D-PAGE, there obviously exists many differences displayed and unmatched protein spots in the 2-D PAGE maps of Pb²⁺ induced rabbit brain. 3 spots were extracted from the 2D-PAGE and detected by the MALDI-TOF-MS and ESI-MS, The experiments show that rabbit brain proteins would be affected by Pb²⁺ injection.

Abstract:Growth inhibitory factor (GIF), renamed as metallothionein-3, is a 68-amino acids brain specific member of the metallothionein family. GIF plays comprehensively physiologic roles and is deemed to be implicated in Alzheimer's disease. When supplemented with Alzheimer's brain extracts, GIF demonstrates its in vitro growth inhibitory activity on cultured neuron; but the underlying molecular mechanism remains unknown. To probe this mystery, the yeast two-hybrid system was employed to screen a human brain cDNA library for GIF-interacting proteins. From 4.1×10⁶ transformants of library screening, a C-terminal fragment of small G-protein Rab3a was obtained, harboring 87 amino acids that interacted with GIF in yeast. The full-length Rab3a was amplified by polymerase chain reaction (PCR) using human total placenta cDNA as template and concomitantly cloned. In yeast two-hybrid test, full-length Rab3a also interacted with GIF. Through coimmunoprecipitation and Western blotting experiments, interaction between GIF and Rab3a was confirmed in mammalian cell. Furthermore, the result indicated that Rab3a interacted with GIF in a GTP-binding form.

Abstract:Using ATP directly synthetical method with ADP and radiophosphate, the effects of azide and inorganic phosphate on the ATP synthesis of F₁F_O-ATPase from pig heart mitochondria are investigated. It is found that P_i not only takes an active part in ATP synthesis as a substrate, but also has an inhibition effect on ATP synthesis of F₁F_O-ATPase. In the presence of 1 mmol/L ADP the inhibition increases with the increase of P_i concentration from 0.01 mmol/L to 10 mmol/L. The viewpoint is different from that azide only inhibites ATP hydrolysis but not affects the ATP synthesis at low concentration (＜1 mmol/L). The results indicate that 0.1 mmol/L azide apparently activates ATP synthesis of F₁F_O-ATPase. The activation is negatively related to the concentration of P_i in the reaction system. When the concentration of P_i is kept at 0.1 mmol/L, with the increase of azide concentration, the apparent activation degree of ATP synthesis by azide is changed. When the concentrations of azide and P_i are equal, the apparent activation by azide is the biggest. The apparent inhibition of ATP synthesis by azide would not begin until the concentration of azide reaches near 0.5 mmol/L. After the concentration of azide is higher than 1 mmol/L, the uncoupling of F₁F_O-ATPase appears. The mechanism of the roles of azide and P_i is discussed.

Abstract:Mitochondria are important intracellular organelles in which energy is generated. Mitochondrial permeability transition pore (PTP) opening will induce mitochondrial dysfunction, then result in cell death. The study was caried out to investigate the influence of green tea polyphenols (GTPs) and five kinds of catechins on mitochondrial PTP opening. The results showed that GTPs and catechins have obvious and different effects on H₂O₂-induced mitochondrial PTP opening. GTPs and its major components EGCG, ECG have inhibitory effects on it, while other kinds of catechins, EGC, EC and (+)-C can accelerate the process. The data provide an alternate interpretation of the potent protective function of GTPs, EGCG and ECG.

Abstract:The nontoxic fragment C of tetanus toxin(TTC) can transport other proteins from the circulation to central nervous system motor neurons. Increased levels of CuZn-SOD are protective in experimental models of stroke and Parkinsons's diseases, where mutations in SOD may cause motor neuron disease. Here the human Mn-SOD is linked to tetanus toxin fragment C gene to construct the fusion gene, then was ligated into prokaryotic expression vector pET-22b（+），expression of the plasmid in E.coli, resulted in the production of a protein has a subunit molecular mass of 71 ku and is recognized by both anti-Mn-SOD and anti-tetanus toxin antibody. The Mn-SOD moiety retains substantial enzymatic activity，where the TTC moiety can delivers the fusion protein to central nervous system neurons. Such fusions should provide a powerful tool for investigating the protective and destructive roles of Mn-SOD in motor neurons.

Abstract:The expression and subcellular localization of protein kinase C alpha and bata Ⅰ in mouse oocytes were studied by immunofluorochemistry and confocal laser microscopy. Double staining was performed to detect the PKC and cortical granules of the MⅡ stage eggs. The results show that both PKCα and βⅠ were expressed in mouse oocyte at GⅤ and MⅡ stages, but their subcellular localization were different. Simple methods of immunofluorochemistry and confocal laser microscopy in mammalian oocyte were developed. By these methods, the expression and subcellular localization can be studied conveniently and precisely, so as to provide an effective tool to the research of reproductive and developmental biology.

Abstract:The optical tweezers are convenient tools for investigation of mechanical characteristic of biological particle. The elasticity of red cell membrane is an important index for people to appraise the ability of blood transporting oxygen. Using single optical tweezers technology, a new method to measure the elasticity of RBC membrane was established. Adopting this new method, the elasticity of normal RBC membrane was detected and were consistent with the results of double optical tweezers method. Moreover, the membrane elasticity for RBC with phenylarsine oxide (PAO) was measured. The result displayed a linear relationship between elasticity decrease and increase of PAO concentration. Therefore, this new method has high sensitivity and is acceptable.

Abstract:Using RACE-PCR technique, a full-length cDNA was cloned from Momordica charantia and named mocPHGPX. The entire mocPHGPX cDNA (AF346906) is 927 bp, and encodes 167 amino acids. The derived amino acid sequence data showed that this protein contains the typical (high conservative) structure of phospholipid hydroperoxide glutathione peroxidase (PHGPX) previously found in animals and plants. Northern blot analysis displayed that the corresponding transcript is abundant in the leaves and stems but lower in the roots. These results will be helpful to make a thorough investigation of the function of plant PHGPX and to understand the anti-oxidative systems of plants in an all-round way.

Abstract:Monocyte chemotactic protein-1(MCP-1) is a potent special chemoattractant capable of recruiting monocytes into the subendothelium. In order to study the patterns of fluid shear stress on induction of MCP-1 secretion in cultured human umbilic vein endothelial cells (HUVEC), the relationship of fluid shear stress and the secretion of MCP-1 was examed, cultured HUVEC were subjected to controlled levels of shear stress (0.4, 1.0, 2.0 N/m²) in a parallel plate flow chamber. MCP-1 in HUVEC of different periods was measured by immunohistochemistry method and image analysis; MCP-1 in perfusion is measured by ELISA. The results of the experiment demonstrated the secretion of MCP-1 regulated by shear stress was time- and force-dependent. The accumulated level of MCP-1 in HUVEC under lower shear stress (0.4 N/m²) for 4～5 h was 3-fold compared with that for static cells. When the shear stress lasted to 6 h, the secretion of MCP-1 was reduced to normal levels and could not be increased. Such research provides data for understanding the mechanism of the contribution of hemodynamic forces to atherosclerosis.

Abstract:Based on a microcomputer installed with the Linux operating system and integrated with the intranet environment, the Blast software with the WEB interface was constructed for visualized sequence similarity analyze. All of the users in the intranet could visit the WEB Blast server using WEB-browser to analysis their sequence easily. The databases to be searched might be either the public database such as EMBL or GenBank nucleic acid sequence database and SwissProt amino acid sequence database or the private sequence databases prepared by themselves. Result demonstrated that this intranet WEB Blast environment is very useful in large-scale nucleic acid sequences and amino acid sequences analysis.

Abstract:Tissue microdissection is potentially one of the most useful techniques in the comparative investigations of specific cells. A novel laser capture microdissection (LCM) has been developed to procure precisely the single cells from complex normal and diseased tissues, in a rapid and practical manner. In this technique, a transparent thermoplastil film(ethylene vinyl acetate polymer)is applied to the surface of the tissue section on a standard glass histopathology slide; a carbon dioxide laser pulse then specifically activates the film above the cells of interest. Strong focal adhesion allows selective procurement of the targeted cells. Now LCM system had been available in proteome analysis and is likely to have a profound impact on proteome analysis.