Abstract:Telomeres are unique DNA-protein complexes at the terminals of chromosomes that play a critical role in protecting chromosomal integrity and in maintaining cellular replicative potential. Telomerase is a specialized reverse transcriptase, composed of both RNA and protein subunits, that elongates telomeric repeats. The changes in telomere length and telomerase activity are closely linked to cell aging and carcinogenesis. Telomere binding-protein may regulate telomere length by regulating telomerase activity, and then control cell aging, immortalization and carcinogenesis.The development of specific telomerase inhibitors will have broad prospect in the aspect of tumor therapy.

Abstract:Loss of telomere DNA repeats in eukaryotic cells is associated with senescence and apoptosis.Activation of telomerase can maintain telomere length stability and make cells immortal.Telomere-binding proteins might regulate the length of telomere by means of regulating telomerase or other relative factors.The progress in study of telomere-binding proteins and the model of telomere length regulation based on it are reviewed.

Abstract:Characteristics of secondary structure, properties of kinetics reaction and possible catalytic mechanism for hammerhead ribozyme were described, the problems to be studied further in action mechanism were also put forward.

Abstract:In eukaryotic cells, messenger RNAs are formed by extensive post-transcriptional processing of the primary transcripts, assembled with a large number of proteins and processing factors in ribonucleo-protein complexes. The protein part of these complexes mainly constitutes a class of about 20 major polypeptides called heterogeneous nuclear ribonucleoproteins(hnRNPs). In addition to previous models that hypothesised a mere structural function, a more diversified and dynamic role for these protein is now proposed. They might actively participate in the events of RNA metabolism and other cellular functions.

Abstract:Rb correlates with tumorigenesis closely, and is an important tumor suppressor gene. Rb protein forms a complicated feedback regulation pathway combined with p16, CDK4/6, cyclinD1 to control cell cycle. Rb is a central step in G1/S checkpoint control to determine cell cycle progression. Rb protein acts as an interface of interaction between nuclear and extracellular signals and is controlled by many extracellular factors which control cell growth and differentiation.

Abstract:Ribulose-1,5-bisphosphate carboxylase/oxygenase(RubisCO) regulates photosynthesis and photorespiration. It is a key enzyme to control the net photosynthesis in plants. It is one of the most abundent soluble proteins in plant also. The enzyme generally exists in plant and some of microorganisms. Some advance in the research of RubisCO was reviewed, It contains the properties、the structure and the function、gene engineering、the activity regulation of RubisCO and its activase.

Abstract:Differential display PCR(DD-PCR)has attracted widespread interest since it was established. It is a sensitive, simple and powerful method to clone differentially expressed gene fragment in different courses of physiology and pathology. But the main drawback of differential display is high false positive and low reproducibility. This restricted its application in life science. Several strategies to reduce false positive of differential display and improve its reproducibilty was reviewed briefly.

Abstract:Apoptosis is an active programmed cell death process during the organism development, cell differentiation and pathological situation. Many studies have revealed that apoptosis is an important and normal part in plant embryo development, tracheary element formation and development of root, shoot, leaf, flower. In hypersensitive response, plants use apoptosis in response to infection by pathogens to protect the whole plant survival.

Abstract:Tranforming growth factor β receptor, divided into five types, are membrane protein. The functin of type Ⅲ receptor is regulating the affinity between receptor and ligand as well as the expression of type Ⅱ receptor. Type Ⅰ and type Ⅱ receptor are functional receptors. They cooperate closely while the signal transduction, and on the other hand. They function differently: type Ⅰ receptor acts as the signal pathway by which transforming growth factor β stimulate the increasing synthesis of extracellular matrix; type Ⅱ receptor has closely relationship to the proliferate and differentiate cells.

Abstract:To study the gene expression profile on nasopharyngeal carcinoma (NPC),lung cancer (LC) and normal adult nasopharynx (NP) tissue and to obtain the NPC-related gene, the total RNA extracted from these tissues were retro-transcripted and labeled with α- ³²P isotype. The cDNA probes were hybrided to high density cDNA microarray GF200 with 5 184 genes and ESTs,then the signals were analyzed by Pathways software. The results showed that the low density signals were distributed in three types of tissues and the number of genes and ESTs with the density value over 200 was respectively 110,134 and 158 among NPC,LC and NP tissue. The results indicated that the different expression genes existed among NPC, LC and NP tissues and some new genes could play an important role in NPC. The high-density microarray was a rapid and effective method to select the different expression genes.

Abstract:Genomic integration and mRNA transcripts of human OSM cDNA in transfected mouse melanoma cells were identified by PCR and RT-PCR methods.A sense primer for the regulatory sequence in carrier vector paired with an antisense primer for cDNA was used in integration analysis, a continuous transcription-unit was amplified with the expected size in OSM cDNA transfected cells but not in the wild type or vector control cells, reflecting a more accurate relationship between integration and expression. In transcription analysis, a sense primer for cDNA paired with an antisense primer for a sequence between the multiple cloning site and polyA signal in carrier vector was used to distinguish exogenous transcripts from endogenous gene products. This method is convenient and specific in determining exogenous gene integration and expression in transfectants.

Abstract:Renaturation of human cilinary neurotrophic factor expressed in recombinant bacteria, by gel filitration chromatograph, was studied. It was showed that the renaturation rate by this way was higher than that by dilution and dialysis, and the protein was purified at the same time. It was a simple, rapid, good reproducibility and efficient procedure which can be used to produce ciliary neurotrophic factor in large scale.

Abstract:The phycocyanin (PC) is a kind of fluorescence molecular probe because it has characteristic absorption spectra and fluorescence spectra.To get highly purified phycocyanin,the successive procedure of column chromatography was set up with SephadexG-200、DEAE-SephadexG-25、HA、and SephadexG-200.The results showed that the purity standral (ratio of A₆₁₅／A₂₈₀) reaches to 14 at pH 7.0,and it has been demonstrated by PAGE as a single band.

Abstract:Using HRP-conjugated goat anti-human apoAI-IgG, an enzyme-linked immunosorbent receptor assay was developed for measurement of high density lipoprotein (HDL) receptors on rabbit liver plasma membranes. A curve of anti apoAI-IgG binding to known amounts of HDL was constructed to quantify the HDL bound. Parallel samples with 25-fold excess goat plasma HDL were assayed to detect nonspecific binding. The results showed that the K_d and B_max of rabbit liver plasma membrane HDL receptors were (7.17±1.18)mg/L and (622.5±146.1)mg/L respectively (n=7).

Abstract:A novel STE20-like protein kinase, Mess1, was cloned from a murine liver cDNA library. The 1.7 kb cDNA encodes a peptide of 497 amino acids. Mess1 is most similar to human MST2 protein kinase with an identity of 95%. A putative kinase catalytic domain, located at the amino terminus of the Mess1 protein, is homologous to that of the STE20 family. There is a serine/threonine and glutamic acid-rich cluster in the carboxyl terminal region of Mess1 that is believed to mediate the binding of SH2 domains. Mess1 might involve the signal transduction on the interaction with proteins with SH2 domains.

Abstract:Total 40 random primers of OPN and OPM groups made in Operon Co. were applied to the RAPD analysis of H.molitrix and A.nobilis. Besides establishment of species-specific RAPD electrophoretogram, some kinds of primers which can produce molecular genetic markers of RAPD with individuality- or population-specificity of these fish were found. A modified method to extracting genomic DNA from peripheral blood cells of fish, and the application prospects of RAPD technique in genetic diversity and variation of fish, and in fishery management and the evaluation of fish resources were discussed.

Abstract:A membrane-bound phosphatidylinositol 4-kinase(PI4K) has been purified approximately 11 500-fold from bovine cerebella cortex. The purification procedure involves: solubilisation of the membrane fraction with TritonX-100, ammonium sulfate fractionation and cation-exchanger chromatography on phosphocellulose followed by affinity chromatography on heparin-sepharose CL-6B. The enzyme was further purified through DEAE-10 FPLC chromatography at last. The purified enzyme exhibited a final specific activity of 450 nmol/mg·min. The molecular mass of the enzyme was estimated to be 56 ku by SDS-PAGE. Kinetic measurements showed that the apparent K_m value of PI kinase for the utilization of PtdIns is 6.6×10^-7 mol/L and for ATP 7.9×10^-7 mol/L. In addition adenosine was found to be a strong inhibitor, Enzymatic activity was found to be stimulated by Triton X-100.

Abstract:To study the regulation of c-fos and c-jun gene expression by introduction of wild-type P₅₃ gene and Rb gene, a P₅₃ gene recombinant adenovirus vector, AdCMV P₅₃, and a Rb gene recombinant adenovirus vector, AdCMV Rb, were transfected into the cultured vascular smooth muscle cells derived from human umbilical artery. The level of c-fos、c-jun mRNA was quantified by reverse transcription-polymerase chain reaction (RT-PCR). The content of c-Fos and c-Jun protein were detected by immunochemical staining. The senescent cells were characterized by β-galactosidase staining. Agarose gel electrophoresis of DNA was used for analysis of apoptosis. The results showed that introduction of wild-type P₅₃ could increase the level of c-fos、 c-jun mRNA and protein, inducing the apoptosis of vascular smooth muscle cells. The introduction of exogenous Rb gene could induce cell senescence and down regulate c-fos gene expression. However, c-jun expression in vascular smooth muscle cells remained constant following transfection with AdCMV Rb.

Abstract:VEGF receptor, KDR, is one of the main VEGF receptors, and it plays important role in VEGF stimulating endothelial cell (EC) proliferation and vascular permeation. To obtain active recombinant KDR which can bind VEGF in vitro, RT-PCR was used to clone KDR (Ⅰ～Ⅳ) DNA fragment from human umbilical vein EC. The cloned KDR (Ⅰ～Ⅳ) fragment was identified with enzyme digestion and sequencing and then cloned into fusion protein expression vector pGEX2T. The GST-KDR fusion proteins were expressed in E.coli XL1-blue after inducing by IPTG. The fusion proteins were extracted from bacterial inclusion body with basic denature method and purified with preparing SDS-PAGE gel followed by electric-elution.

Abstract:The data-processing ability of double wavelength/double beam spectrophotometer has been greatly enhanced by connecting it to a computer and designing new software. Formerly, the instrument relied on an old recorder to plot curve on a special-designed recording paper and had poor data-processing ability: user had to determine manually the abscissa and ordinate for any data point in the curve with a ruler, which limited its use in biological studies, especially in kinetic analysis. Currently a computer and a software packet designed are applied to it, which leads to the following several distinct advantages over the old type: sampling and storing data by computer, showing more than one curve simultaneously on the screen, showing the value of data points, determining the positions of peaks automatically, smoothing curves, zooming in or zooming out graph, printing graph with a HP Laser Jet. The structure of the new system is introduced here, based on which the effect of phosphatidylethanolamine content on the Ca²⁺ uptake of Ca²⁺-ATPase is studied.

Abstract:The chemiluminescence behaviour of pyrogallol alkali autoxidation produced O₂^-_· was studied. The optimum conditions were investigated by examining the influence of various factors. The improved method needn't to use the luminous agent——luminol, the reagents used are cheap, both the stability and reproducibility are good, and the sensitivity is far higher than the report of other method of pyrogallol autoxidation.

Abstract:The cytoplasmic domain of human erythrocyte band 3 (cdb3) plays an important role in connection of the membrane with the membrane skeleton, and environment inside the cell as well. With whole length of band 3 gene, cdb3 gene was amplified by PCR and then was cloned into pRSET plasmid. The plasmid was introduced into E.coli BL21(DE3). The cdb3 expression proteins were purified and its function was verified according to the ability to inhibit aldolase.

Abstract:A high level expression vector pRC has been constructed, which contains P_R promoter, cIts857 gene, multiple cloning sites(MCS) and two strong transcription terminators. With this vector, hTNF-α, hIL-3 and anti-HEL McAb Fd genes have been expressed successfully, and their products each account for more than 36% of the total cell proteins. At the same time, it has been studied whether such factors as procaryon enhancer sequence and different strains affect the expression level of P_R promoter vector. Furthermore, the inductive efficiency was compared among P_R, P_L and P_RP_L promoter, and the result has showed that P_R or P_L is as strong as P_RP_L.

Abstract:A U937 cell model, in which Bcl-2 expression was suppressed, was established by transfering antisense bcl-2 gene into U937 cell. The model cell had normal growth and survival. Crystal violet assay with actinomycin D and ELISA indicated that the activity and concentration of TNFα in the model cell supernatant had no change, but mouse thymocyte proliferation assay revealed that IL-1 activity increased in the supernatant siginificantly. The results suggested that Bcl-2 expression has no effect on TNFα production and its activity, but it may suppress the expression or activity of IL-1. The study on the regulation of IL-1 and TNFα expression by bcl-2 may provide some beneficial data for elucidating the mechanism of bcl-2 action.

Abstract:An engineering anti-human TNF-α single-chain antibody (ScFv) gene was cloned into the expression vector pET15b-Etag, and expressed in E.coli BL21(DE3) under the control of T7 promoter. By using 0.1 mmol/L IPTG induction, the amount of the ScFv expression product was more than 38% of total bacterial proteins. Most of them existed in a form of inclusion body. More than 6% of total bacterial proteins were soluable and can be detected in the part of periplasm, which can bind with rhuTNF-α by ELISA and Dot-blotting technique.

Abstract:Ultra-weak chemiluminescence analyzers that detect weak light from samples were developed by ultra-weak chemiluminescence analytical technology. BPCL ultra-weak chemiluminescence analyzer has a lot of supper performance. It is satisfactory to research and application in the fields of biology, medicine and chemistry.The example that of to research DNA damage was introduced. It is approved that the technology and analyzer were advanced and practical.

Abstract:DNA microarray or chip is a new technique in molecular biology.With combining the light-directed chemical synthesis, semiconductor-based photolithography,and solid-phase chemical synthesis, thousands of oligonucleotide probes were arrayed or spotted on the surface of solid support. This technology has successfully monitored the simultaneous expression of many thousands of genes, developed to screen DNA mutation and polymorphism, applied to sequence DNA, and discovered the novel disease-related genes by hybridization to radioisotope or fluorescence labeled DNA or cDNA coming from interesting tissues and cells.

Abstract:L-fucose (L-FC),6-deoxygalactose, is one of rare sugars which naturally exists in the form of L-isomer. L-fucose plays an important role in their physiological and biological functions. It has been well known that L-FC is attached to glycochains of tumor markers such as alpha fetoprotein (AFP) and CA-199. Increment of α-L-fucosidase was detected in patients with primary hepatic carcinoma. Fucolipid accumulation in human adenocarcinoma was abnormally increased and fucosyltransferase was highly active in lung cancer cells. In addition,it has been found that patients with liver cancer and cirrhosis excreted free L-fucose via urine to greater extent than normal individuals. The values of UFC were detected in the urine from 86 health people and 205 people with various tumors. The mean value of urinary L-FC in health people was 177.7 μmol/g.cr(s=56.6, 95% confidence range was 63.8～290.2 μmol/g.cr). The mean values of UFC in urine obtained from patients with hepatocellular carcinoma, cirrhosis, acute and chronic hepatitis, pancreas carcinoma, gastric cancer, lung cancer , intestinal cancer, esophageal cancer were higher than 290.2 μmol/g.cr, having significant difference from normal individuals. The mean values of UFC in urine obtained from patients with mastadenoma, cervical cancer and ovary cancer have significant difference from normal individuals. The value of L-fucose in urine can be regarded as one of the characteristic indexes for hepatocellular carcinoma and canceration of acute and chronic hepatitis. Detection for L-fucose is one of tumor marker tests for patients with tumor in digestive system.