Abstract:Development and morphogenesis of visual system is a complicated procedure, and the gene regulation is widely paid attentions by neurodevelopmental biologists. Vax (ventral anterior homeobox) family was found in 1998, which is related to development of forebrain, optic vesicle, eye-predium, optic stalk and neuroretina. Vax-1 is involved in differentiation of pigmented epithelium and optic stalk, while Vax-2 could function in the establishment of the dorso-ventral axis of the retina and visual system. Investigation of Vax genes will be helpful to understand the mechanism of molecular events, which happened during the early development of the visual system.

Abstract:Iris recognition, an emerging biometric technology, which exploits the uniqueness of human iris texture, can accomplish automatic personal identification. Of all iris recognition systems so far, Daugman’s system based on Gabor wavelets encoding iris texture is more robust. The principle of iris computer recognition was reviewed in two parts, the principle of encoding and the principle of statistical strategy.

Abstract:Telomeres shortening during proliferation of many cell lines can not be explained completely by the mechanism of the end replication problem. Under 40% oxygen partical pressure, cell proliferation is blocked and the rate of telomere shortening is increased. Blocked cells present senescent characteristics and accumulated single-strand breaks in telomeres. It might be speculated that accumulation of single-strand breaks and resultant loss of distal single-stranded fragments during replication could be a major cause of telomere shortening under the condition of senescence or oxidative stress. The precise mechanism of the positive or negative regulation on the telomere length by telomerase and reactive oxygen species still remains unclear.

Abstract:Catalytic antibody (also known as abzyme) is a kind of immunoglobulin with catalytic activity. Because it has high selectivity and amazing diversity as antibodies and highly catalytic activities as enzymes, it is anticipated that using abzyme preparation technology one can obtain any kind of tailor-made biocatalysts, including those not occurred in nature, for various kinds of practical applications. Thus, abzyme study has an important value in theory and practice for biology, chemistry and medicine etc. The new advances of catalytic antibody research are summarized with a special emphasis on the breadth and scope of new antibody-catalyzed reactions and novel hapten design strategies. The problems to be solved in this field are discussed and the strategies for solving these problems are presented with special focus on increasing the catalytic activities of abzymes.

Abstract:Artificial evolution has emerged in the past few years as a powerful alternative to rational approaches for engineering protein. In vitro evolution of enzymes use error-prone PCR , DNA shuffling and mutator stains. Many useful enzymes have been isolated following the artifical evolution.

Abstract:There are two kinds of changes of gene expression,that is,novel gene expression and differential gene expression in quantity. Cloning technique of differentially expressed gene in quantity mainly is mRNA differential display, which is presently one of the most effective methods. However, mRNA differential display possesses higher unreal positive rate,in order to overcome its shortcoming, some novel strategies and methods were advocated,such as differential subtraction display, subtraction based on LD-PCR, LD-PCR based on subtraction,those techniques have dominant advantages to mRNA differential display.

Abstract:Tissue engineering was brought forwards only ten years. It has gotten high recognition all over the world and has been regarded as “a new growth point of economy” by many country. They have invested a lot of money as well as people and have gotten a lot of harvest. In China, many research groups are doing the related studies too. The second bi-annual meeting of the tissue engineering society was held in Florida, USA and the newest results of tissue engineering research were delivered. Here summarized its research development and put forward that how to ensure the function of engineered tissue is a very important question.

Abstract:Trefoil factor family (TFF) peptides have a special domain called trefoil domain. Trefoil domain contains highly conserved cysteine, arginine, glycine, tryptophane, phenylalanine and a unique three-loop structure which is formed by intrachain disulfide bonds between six conserved cysteine residues in the 1～5, 2～4, 3～6 position. Three most important TFF peptides are TFF1/pS2, TFF2/SP (spasmolytic polypeptide) and TFF3/ITF (intestinal trefoil factor). The ectopically expressed sites of them are body and fundus of stomach, deep foveolar pits of gastric antrum and goblet cell of small and large intestine, respectively. TFF may play an important role in both maintaining the barrier function of mucosal surfaces and facilitating healing after injury. The structures of TFF peptides are very compact and contain α-helix，β-sheet. The mechanism of TFF is not clear and two hypotheses were proposed which were co-working with mucin or receptor.

Abstract:Ras-GTP-Raf complexes play an important role in the RTKs-mediated Ras pathway. The molecular mechanism of signal transduction in the Ras-GTP-Raf complexes is that the complexes switch signal transduction, trigger signal amplification cascade and may be involved in neoplasia. In addition, Ras-GTP-Raf complexes are essential in the PKC-mediated activation of Raf.

Abstract:Cytokines of the TGF-β family exert multiple biological effects by their signal transduction pathways that undergo a basic course as follows: ligands of TGF-β family→receptors→SMADs→transcription factors→DNA expression. In stimulating their own two type receptors with kinase activity the cytokines first bind to type-Ⅱ receptors and then the ligand-binding type-Ⅱ receptors activate type-Ⅰ receptors. The type-Ⅰ receptors phosphorylate pathway specific SMADs. These activated SMADs then associate with common SMAD and translocate from cytoplasm to nucleus where they regulate transcription either by associating with transcription factors and by binding directly to DNA.

Abstract:In order to establish warning system of gastric cancer, using cell line GC7901 and GES-1 as targets,8 cDNAs of differentially expressed genes between them were isolated by mRNA differential display,and then respectively named as GCYS-1 to GCYS-7,GCYS-20,and then were cloned into pGEM-T vector. These cDNA fragments were up-regulated in GC7901 and down-regulated in GES-1 as shown by Northern blot.DNA sequencing demonstrated that all of them were novel.These sequences have been assigned the database accession numbers in GenBank as below:AF054162,AF054163,AF054164,AF054165,AF054166,AF054167,AF054168,AF219140.

Abstract:The molecular dynamics research of the core domain of p53 protein crystal structure shows that besides the stability in biochemistry this domain also shows a high stability in molecular mechanics. Based on that work, the residue R249 was substituted with amino acids Gly and Ser respectively, and molecular dynamics researches were performed separately. The results show that these substitutions cause a relax tendency between loop2 and 3 domains, leading to an alteration of the whole conformation of p53 core domain and ruining its stability. The results visually explains the mechanism of p53 changes in immunological and biochemical reactions, which are caused by 249 residue substitutions from 3-D structure variations.

Abstract:A method and a device were developed to monitor biomass concentrations on-line and in-situ in a fermentation process. The presence of microbial cells would affect the dielectrical properties of microbial suspensions. At radio-frequencies the dielectric permittivity of cell suspensions are monotonic function of the measuring frequency and cell density. Based on this understanding a new method for measuring biomass concentration is proposed. Using this method the biomass concentrations are measured on-line and in-situ without taking samples from the bioreactor, and only viable cells are detected. The electrode could be directly inserted into the fermentor and could be sterilized in place. The method has a wide use in the industrial field of pharmacy, brewery, sewage disposal.

Abstract:The expression vector, pET21-CEA-streptavidin was constructed which contains the gene of single chain antibody of CEA(ScFv) and the streptavidin. Constructed vector was expressed by Escherichia coli. The results showed that such protein was successfully prepared and its bispecific activities were testified by affinity blot and Western blot. The biotin binding and immunocytochemistry assay indicated that the expressed fusion protein was capable of binding biotin molecule and CEA antigen respectively. This protein has the potential application when combined with the corresponding tests.

Abstract:To establish a flow cytometric method (FCM) for simultaneous evaluation of asialoglycoprotein receptor (ASGPR) on the surface of normal rat hepatocytes, injured rat hepatocytes and hepatoma cells (BEL-7402). FCM for ASGPR was established using normal hepatocytes (L-02) and FITC-conjugated galactosyl-neoglycoalbumin (FITC-NGA), the specific ligand for ASGPR. The mean intensity of fluorescence (MIF) of the three different hepatocytes were determined and calculated after simultaneously incubated with FITC-NGA at the same concentration. The concentration of FITC-NGA for saturating ASGPR on the surface of L-02 was 0.4 mg/L, at which the MIF of the three different hepatocytes were 228.7，5.81 and 1.13 respectively. The saturated combination can be completely inhibited by 50-fold NGA or 10mmol/L EDTA. FCM can display soundly the characteristic of the receptor-ligand combination between ASGPR and FITC-NGA. Compared with normal rat hepatocytes, there is no ASGPR on the surface of BEL-7402, and the quantities of ASGPR on the surface of injured rat hepatocytes decreases significantly.

Abstract:The change of conformation and activity of human placental alkaline phosphatase (PLAP, E.C.3.1.3.1) in pentachlorophenol (PCP) solutions of different concentrations were studied by enzymic activity measurement and fluorescence spectra. The inhibition type of PLAP by PCP and the effect of pH on the enzyme inhibition were simultaneously measured. In the PCP concentrations lower than 1.0 mmol/L, the enzyme activity and fluorescence intensity rapidly decreases with a marked red shift of emission maximum. In the PCP concentrations higher than 1.0 mmol/L, the enzyme activity and fluorescence intensity gradually decreases with a continuous red shift of emission maximum.At 5.0 mmol/L PCP, the enzyme intrinsic fluorescence quenched and the enzyme activity is 51.4% of original activity. At 10.0 mmol/L PCP, the enzyme residual activity is 30.0%. PCP is an uncompetitive inhibitor of PLAP. The inhibition constant(K_i) is 3.86 mmol/L.The activity inhibition of PLAP also affected by the pH, its activity inhibition disappeared below pH 7.5,but gradually increasing between pH 7.5～10.5. The results suggest that the activity inhibition and the conformation changes of the enzyme were caused by PCP. The inhibition of PLAP activity correlates with the dissociation state of PCP.

Abstract:Immobilized metal-chelated affinity chromatography(IMAC) membranes were prepared for separating a recombinant fusion antibacterial peptide,which carried a polyhistidine sequence (HIS₆-tag) at the N-terminus. Low bleeding of metal ion was achieved. It was proved that the properties of IMAC membranes were better than conventional chelating sepharose fast flow column. The fractionation of recombinant proteins that carry a polyhistidine tag is currently perhaps the most promising application of IMAC.

Abstract:The key process and information gateway of ischemia/reperfusion injure after improved by drugs and way were researched. Langendorff installation and cardiac muscle of Wistar rats were used. The excitant of adenosine A1 recepor R-PIA was selected as protecting medicine; ATP sensitive potassium channel retarder glybenclamide and PKC inhibitor stauroprine were used simultaneously or separately. The changes of oxygen free radical, nitric oxide, ATPase and inducible nitric oxide synthetase gene were observed, which were compared with ischemia pretreatment at that time. The results showed that R-PIA and ischemia preconditioning had preferable protection and these effects depend on the opening of potassium channel and the activing of PKC. The opening of potassium channel depends on partly the activing of PKC, but opening of potassium channel may be a key factor even more than PKC at downriver position.

Abstract:Venom of sea anemone Anthopleura qingdaoensis contains at least two cardiotonic polypeptides capable of enhancing cardiac contraction(a positive inotropic effect). An artificial gene encoding anthopleurin-QD2 (Ap-QD2), which consists of 49 anino acid residues, was cloned and transferred into pPICZαA, a secretory expression vector for Pichia pastoris. The construct was linearized and was integrated into the yeast chromosome by electroporation under the selection of zeocin. Approximately 20 mg/L of biologically active Ap-QD2 was produced from one of the KM71(Mut^s) transformants, and about 7 mg of pure Ap-QD2 was obtained after chromatography purification.

Abstract:A water-soluble carboxymethylated derivative （CM-HNP） of Hunai polysaccharide （HNP）from the sclerotia of Pleurotus tuber-regium （Fr.）Sing. was prepared by the reaction of HNP with monochloroacetic acid. The antioxidative activities of CM-HNP were also studied. The results showed that CM-HNP can protect liver mitochondria from lipid peroxidation induced by Fe2+-Vit C including the increase of TBARS contents, the swelling of mitochondria and the decrease of membrane fluidity with a dosage-effect manner. In addition, CM-HNP can effectively scavenge O-·2 generated by the self-oxidation of pyrogallic acid.

Abstract:In order to define insulin-like growth factor 1(IGF-1) binding sites on IGF-receptor 1 (IGF-R1), the interactions of IGF-1 and IGF-R1 was verified in the yeast two-hybrid system. Seven mutants IGF-R1 were generated by site-directed in vitro mutagenesis, then the interactions of IGF-1 and various mutants IGF-R1 were quantified to identify the gain-of-function mutation in the yeast two-hybrid system by assaying the β-galactosidase activity. The results confirmed the binding data of IGF-1 to IGF-R1 and suggested that the residues N237 and T238 on IGF-R1 play a crucial role in binding to IGF-1.

Abstract:According to the surface electronic potential distribution of the compound hIL-6/hIL-6R and gp130 was analyzed by the program Delphi, the space conformation of the tertiary compound hIL-6·hIL-6R·gp130 is studied with Docking method. The stable structure of the tertiary compound is obtained during the optimization with molecular mechanics and molecular dynamics at normal temperature. The binding domain of the stable compound was discussed by intermolecular force(including Van der Waals force, hydrogen bond, salt link, etc) and reaction free energy theory. The results showed that the surface of gp130 enriches negative charges whereas the compound hIL-6/hIL-6R positive charges, gp130 combines hIL-6/hIL-6R with surface electrostatic interaction for transduction signal, helix-C, loopBC, loopCD in hIL-6 participate in the interaction with the regions loopEF,linker, loopA′B′, loopB′C′, loopD′E′in gp130, and the regions loopA′B′, β-strand E′ in hIL-6R are interacted with the regions loopA′B′, β-strand E′ in gp130.

Abstract:To purify the telomerase of HeLa cells and to study its protein components. The method of affinity purifying was used to extract the telomerase component from the crude extract of HeLa cells, basing on the specificity of telomerase containing protein and RNA. And then the activity of telomerase was tested, SDS-PAGE was used to test protein components. There four bands were observed in the SDS-PAGE gel, there are two near 212.2 ku, one near 97.4 ku, one near 42.7 ku compared with the protein mark. It was shown that the product with telomerase activity was obtained using the affinity purification method.

Abstract:Four chickens, each from a different strain, were analyzed using randomly amplified polymorphic DNAs(RAPD) technique with 12 primers. 38 of 99(38%) detected fragments showed polymorphic. Four individual-specific fragments were recovered and probed with labeled whole genome of chicken. Positive signal was detected in three of the fragments, suggesting that hypervariable fragments generated by RAPD contain repetitive sequences. Highly variable DNA fingerprints were generated when such repetitive sequence containing fragments were used as probe to hybridize to HaeⅢ-digests of random chickens’DNA. Therefore, RAPD products that are highly variable among individuals can be used as DNA fingerprinting probes.

Abstract:Sphingosine-1-phosphate(SPP) is an important second messenger involved in cell growth and cell death. HEK293 cells transcripted with EDG-1, a SPP receptor, were cultured and harvested, and incubated with ³²P labeled or non-labeled SPP. The SPP level was determined based on competitive binding of SPP or ³²P-SPP to HEK cells. This method does not require special apparatus, and able to measure the content of SPP as low as pmol with high sensitivity and good repetition. The between groups deviation was less than 15%.

Abstract:Angiogenesis-related diseases involving EGF include acherosclerotic plaques, haemangioma, angiofibroma, tumor growth and arthritis. EGF may serve as a drug target and its antagonists may have important clinical applications.Peptide phage display libraries have been successfully applied in areas of finding ligands for enzymes, receptors, and many other molecules. A pⅧ-based peptide phage display library was panned with the cytokine EGF and several EGF-binding clones were selected based on ELISA and micropanning assays. The selected EGF-binders from peptide phage display library may be utilized in affinity chromatography in EGF downstream processing and even act as potential antagonists of EGF if their affinity is further improved through secondary library strategy.

Abstract:The polymer of the urushiol tyrosine resin coordinating Cu²⁺(UTR-Cu²⁺) was synthesized to immobilize polyphenol oxidases.The results of experiment showed that the method was practicle. Optimum pH of the immobilization polyphenol oxidases was 6.64 and 7.17. The effect of the temperature on UTR-Cu²⁺-enzyme was discussed.Michaelis constant was determined and compared with the free enzyme. It was suggested that the polyphenol oxidases was immobilized by coordination bond, and the model of immobilization enzyme with UTR-Cu²⁺ was proposed.

Abstract:Mediated by PMS,the NADH resulted from the reaction catalysed by LDH isozymes can reduce K₃Fe(CN)₆ into K₄Fe(CN)₆. Then K₄Fe(CN)₆ can react with FeCl₃ resulting in Prussian blue which dyes LDH isozymes.

Abstract:Ultra-weak chemiluminescence analytical technology has been developed rapidly in recent years. The technique in pharmaceutical analysis was introduced by using two examples. One is based on the chemiluminescence reaction of Ce(Ⅳ)-analgin-rhodamine 6G and the other is based on the auto-oxidation of analgin in micellar medium. An ultra-weak chemiluminescence analyzer has been coupled to flow-injection device for the determinations.

Abstract:dbEST is an important division of Genebank. EST clones represent useful molecular tools for new gene identification, gene expression and recombinant protein expression studies. A description of dbEST and the way to get the best of it were given.

Abstract:Synthetic oligonucleotides and their analogs have shown promising potential therapeutic agents in the treatment of viral infections and certain cancers. As drugs, the full characterization of oligonucleotides requires complete verification. Chromatographic or electrophoretic techniques can be used in analysis of concentration and purity of synthetic oligonucleotides, but the analytic methods have been limited in analysis of base composition, sequence, identity and modification of oligonucleotides. Ability of mass spectrometry to discriminate mass make MS effective, sensitive, rapid and accurate to determine these characterizations of oligonucleotides. application of mass spectrometry in quality control of synthetic oligonucleotides was introduced.

Abstract:The agglutinin precipitation assay was used to determine the bone alkaline phosphatase. The advantage of this assay is simple manipulation, and good reproducibility. Coefficient variation within and between runs were 6.12%, 8.5% and 6.4%, 9.5% respectively. It is concluded that serum B-ALP levels is an useful parameter for clinical observation of the bone metabolism.