Abstract:Matrix association region (MAR) is not only a kind of cis-acting element for regulation of eukaryotic gene expression and chromosome dynamics, but also a type of fixed and specific DNA marker on the eukaryotic chromosomes. MAR library can be constructed by in vitro or in vivo association of DNA fragments with the nuclear matrix. In vitro MARs can be used in construction of chromosome's physical map, and in vivo MARs can be applied in studying the expression, regulation and chromosomal organization of known genes, and analyzing the newly discovered genes.

Abstract:Chemokines family is divided into four subfamilies(CXC、CC、C&CX3C)，it has been estimated that there may be as many as 40 to 50 human chemokines. Structure,gene location and function of chemokines and their receptors have gradually been eluciated. They play important roles in normal and disordered physiological states.

Abstract:Molecular beacons are hairpin-shaped oligonucleotide with a fluorescent moiety and a non-fluorescent quenching moiety attaching to each ends and can be used to report the presence of specific nucleic acids in homogeneous solution or in living cells.The design principles and advantage of molecular beacon and the application for gene mutation detection were briefly introduced.

Abstract:Aromatase plays an important role in the initiation and development of breast cancer. A full description is given on the structure, the unique tissue-specific expression pattern , the factors and mechanisms involved in the expression regulation of the gene of aromatase，CYP19.

Abstract:Mitogen-activated protein kinase (MAPK) cascades play essential roles in the signal transductions in eukaryotie. The MAPK cascades are composed of MAPK (mitogen-activated protein kinase), MAPKK (mitogen-activated protein kinase kinase) and MAPKKK (mitogen-activated protein kinase kinase kinase). These kinases are structurally conserved, and transduct signals via protein phosphorylation. Many kinases and genes of yeasts, animals and plants which are involved in the MAPK cascades have been isolated and characterized. From these investigations, the signal trasduction pathways of eukaryotic cells gradually become clear.

Abstract:Gene knockout is a powerful technique to elucidate the molecular mechanism of learning and memory, this molecular-genetic approach brings exciting new improvement in memory research. A number of gene knockout mice which have defects in learning,memory and long-term potentiation, long-term depression have been generated and it was found that lots of genes are necessary in the process of learning and memory.However, overlooking the role of background genes is a major problem in the present studies, the phenotypical abnormalities attributed to the targeted gene may be simply result from the effects of background genes.In order to overcome this limitation, it is necessary to develope new ES cell lines and use inbred mouse strains from pure background, furthermore, methodolgical details must be improved and fine control over the timing,locale and degree of genetic disruption must be gained.

Abstract:Aquaporins, water specific conducting channels, ubiquitously exist among animals, plants and microbes. There are six kinds of aquaporins in mammalian plasma membrane, which locate on the organs that participate actively in water metabolism. Plant aquaporins exist both in plasma membrane and tonoplast, which have a general role in regulating transmembrane water transport during the growth, development, and stress responses of plants. Most information about the structure and function of aquaporins comes from those researches on AQP1 that exists in erythrocyte membrane. Aquaporins assemble in the membrane as a homotetramer with each monomer having its individual water conducting function.The distribution of tetramer in membrane is unsymmetrical, which exhibits four protrusions in the inside surface of membrane and forms a large central cavity outside.

Abstract:RET proto-oncogene was rearranged during transformation,and so has the name.It encodes a cell membrane receptor tyrosine kinase.In recent years,the possible correlation between human diseases and Ret activation by mutation,such as rearrangement,point mutatoin,and deletion were studied.Its rearrangement with other gene fragment was frequently found in thyroid papillary carcinoma tissues.Point mutations were detected in multiple endocrine neoplasia 2 and familial medullary thyroid carcinoma.Hirshsprung syndrom may be resulted from its deletion. The structure and possible function of RET gene,its mutation effects to various human diseases were summarized.

Abstract:Microchips are constructed with photolithography technology and chemical synthesis. Microchip-based devices can be applied for separation, preparation and preconcentration of biological sample. The devices are also used for polymerase chain reaction or ligase chain reaction as the reaction vessels. The most attractive application is that the microchip immobilizing various oligonucleotide arrays can be used to detecting gene mutation. In addition, these compact arrays of probes may be used for ultrafast DNA sequencing. The fabrication, principle, characteristic and application of microchips were introduced.

Abstract:Disulfied-stabilized Fv (dsFv) fragments are new type of engineering antibody, in which varible regions of heavy chain (V_H) and light chain (V_L) are stabilized by an interchain disulfide bond. dsFv is more stable, an important biochemical feature for the preparation and handling of these proteins in clinical setting.

Abstract:The amplitudes and the phases of first to sixth harmonics of P-VEP waveforms were calculated, thus 12 parameters of the P-VEP waveform were obtained for the discriminatory analysis. The P-VEP waveforms of thirty-two normal eyes, thirty-one amblyopic eyes and thirty eyes with retrobulbar neuritis (RBN) were used to establish the discriminatory system. In the test of the discriminatory system, thirty of the thirty-two normal P-VEP waveforms were classified into normal group, thirty-four of the thirty-five amblyopic P-VEP waveforms were classified into amblyopic group, and twenty-eight of thirty RBN P-VEP waveforms were classified into RBN group.

Abstract:Two kinds of artificial ligand of asialoglycoprotein receptor (ASGPR), galactosyl human serum albumin (Gal_nHSA) and galactosyl poly-L-glutamic acid (Gal_nPLGA), were synthesized chemically. IC₅₀ of synthetic ligands inhibiting the binding of ¹²⁵I-asialofetuin (¹²⁵I-ASF) to ASGPR of rat liver cell membrane were assayed. The results showed that Gal₁₂HSA, Gal₁₅HSA, Gal₂₆HSA, Gal₃₀HSA and Gal₃₄-PLGA could all efficiently inhibit the binding of ¹²⁵I-ASF to ASGPR. The affinities of Gal_nHSA increased along with the increase of the coupling number of galactosyl. Here, the synthesis and the affinity identification of Gal₃₄PLGA with ASGPR were firstly reported. These synthetic ligands with abundant sources and simple preparations may be used as targeting ligands for the liver-targeting delivery of drugs or genes.

Abstract:The cDNA of human brain-derived neurotrophic factor (hBDNF) and the fragment encoding mature hBDNF were amplified from human genomic DNA by PCR, and separately ligated into pUC18. Two inserted fragments were analysised by sequencing, which are correct.The expression of the hBDNF was controled by CMV promotor in NIH/3T3 cells transfected with recombinant plasmid, and analysed by RT-PCR, showing existance of BDNF mRNA in cells. The expression product of the mature hBDNF under the control of T7 promotor was in the inclusion body of E.coli, and visualized by SDS-PAGE analysis.

Abstract:Water content is a crucial factor influencing the enzyme activity in organic solvent.The techniques of water control were investigated in the esterification of Naproxen catalyzed by lipase from Candida cylindracea in isooctane/octanol system. The results demonstrated that the salt hydrate pair —— Na₂SO₄·10H₂O/Na₂SO₄ had the capability of bufferring water. The sensibility of enzyme to water was decreased by adsorbing the lipase on celite —— an apolar carrier. In addition, Esterification could be improved effectively by adding molecular seive to remove the byproduct-water.

Abstract:A new highly sensitive HPLC method for determination γ-aminobutyric acid(GABA) in serum was developed by precolumn derivatization of GABA with 9-fluorenylmethyl chlorormate(FMOC-Cl) using ε-amino-n-caproic acid as an internal standard. The column used was Shim-Pack CLC-ODS(M), 4.6 mm×150 mm,5 μm. Mobile phase A consisted of sodium acetate buffer (0.05 mol/L,pH 3.65)/water/tetranhydrofunan/glacial acetic acid(250/100/15/2.2).Mobile phase B was acetonitrile/methanol (4/1). The conditions of derivatization and chromatography were studied to find a optimal procedure,which included pH value,reaction time, ionic strength and amount of derivatizing reagent.The coefficient of variance of the method was less than 4.6% for within runs, 6.1% for between runs, respectively. The minimal detection limit (S/N=2) was 3.1 nmol/L. The linearity was observed from 10 to 1 000 nmol/L, and the coefficient of determination was 0.9992. The average recovery was 97.1%.

Abstract:A multiphasic buffer system was used to seperate the human short tandem repeat DNA fragments.In stacking gel the main content of buffer were bistris,sulfuric acid and bicine while tris, sulfuric acid and bistris play a major role in seperating gel.The DNA fragments,which can be stacked efficiently in stacking gel and destacked completely in seperating gel, were seperated according to their size. By this way the high-resolution results can be achieved.

Abstract:Recombinant human IFN-γ from Chinese Han nationality,overexpressed in E.Coli, was found to accumulate in cytoplasmicinclusion bodies.After washing, the inclusion bodies were dissolved in 8 mol/L urea. Under the denatured states, the recombinant human IFN-γ from Chinese Han nationality was purified by size-exclusion chromatography and ion-exchange chromatography.The final yielded product was of high purity(96.87%) and exhibited the mass of molecule 17.32 ku analyzed by mass spectrometry.The renatured IFN-γ, which was refolded by diluting, had specific antiviral activity of 5.5×10⁵ U/mg. The sequence of 16 amino acid residues from NH₂-terminus of the protein was determined and was found to agree with that of hIFN-γ reported by Gray. The composition of amino acids of the protein was analyzed. The result had good agreement with that deduced from cDNA of IFN-γ from Chinese Han nationality.

Abstract:By utilizing imported chemiluminescent reagent, self-prepared rabbit anti-human type Ⅳ collagen IgG and a horseradish peroxidase-labeled anti-human type Ⅳ collagen monoclonal antibody, a method of sandwich chemiluminescent enzymimmunoassay(CLEIA) was established then evaluated. The results showed the CLEIA has a lower detecting limit of 30 μg/L，the average recovery was 94.5％, CVs of intra-assay and inter-assay were 6.0％ and 11.6％ respectively, the CLEIA method has a good correlation with Japan kits (ELISA, r=0.9686). the serum reference level by testing 60 cases health is 160 μg/L. It was found that the CLEIA is a sensitive, acurate, stable and specific method，it may be a qualified candidate for serum type Ⅳ collagen detemination of non-radioactive method.

Abstract:The whole designed gene expressing a peptide consisting of calcitonin gene-related peptide (CGRP) (37aa) itself, a signal peptide at its N terminus and an amidated site at its C terminus is cloned into mammalian expression vector pBPV. The pBPV-CGRP plasmid DNA is directly injected into the spontaneous hypertension rats (SHR) and results in the significant decrease of SHR's blood pressure during 3～22 days after injection. The percent of decrease reaches more than 12%, even up to the top 24.42% on the 16th day. The effect can last for longer time and also be better than that of CGRP protein.

Abstract:The change of quality and quantity of wound fluid during different stages could reflect the course of wound healing in a certain degree. The effect of WF taken from different post wounding days on the proliferation of mouse wound fibrolast(mWFb) was studied, and the regulatory role of wound microenvironment on the growth of mWFb was also investigated. It was performed in two kinds of culturing medium: 1640-1%FCS-10%WF. It was found that WF obtained from postwounding days 1,3 and 7 could effectively stimulate the proliferation of mWFb, while, WF obtained from postwounding days 9,11 and 15 with 10%FCS presented inhibitory effects on the proliferation of mWFb. However, the same WF with 1%FCS may induce death of mWFb. The results showed that wound microenvironment could stimulate the proliferation of mWFb within the first week after wounding, and that cell growth may be arrested by WF of the wound during the second week. These might be some growth inhibitory factors in the wound microenvironment of later stage during wound healing.

Abstract:Genomic DNA extracted from the posterior silk glands of two silkworm varieties,C108 and Dazao were digested with PstⅠ and ligated with synthetic adapter.Selective amplification of DNA fragments （SADF） was performed using suitable primers which annealed to adapter regions.Agarose gel electrophoresis shown that DNA polymorphic fragments were appeared for both C108 and Dazao.