0.05), which were lower than that with manual compression. The failure rates were 8.3% and 9.2% respectively, but the vascular complications were less than 1%. Hematoma and femoral artery infection were seen in the PCI group, but happened less compared with manual compression. The removing of the arterial sheaths will not be limited by the anticoagulation and antiplatelet therapies, and it could be performed immediately after CAG and PCI, making hemostasis more easy to archeive. It could also reduce the burden of the medical staff, and be accepted by the patients. Conclusion Femoral arterial closure following PCI using regular and intensive anticoagulation and antiplatelet therapies could be safe and effective with vascular complication rates similar to or lower than with manual pressure.

Balb/c mice were immunized by intraperitoneal iniection of group A erythrocytes. Immunized spleen cells were fused with Sp2/0 murine myeloma cells from the solid tumour. After 10～15 days of incubation at 37℃ CO_2, the supernatants were screened for the agglutinabal antibodies. Three hybridoma cell lines secreting high titer and specific monoclonal anti-A antibodies were established.These hybridoma cells have the ability of constant seceting antibodies which belong to the IgM class. The specificity monocloal anti-A sera was the same as the human anti-A sera.

This article introduces a new educational model-Ability Standard Teaching Mode,which emphasizes that students should participate in teaching and promotes the students to change their thinking style from memory and imitation style to thought and creativity style.

Objective To fractionate phosphoproteome of mouse liver by two-dimensional (2D) liquid phase chromatography fractionation. Methods Phosphoproteins were extracted from lysates of normal mice livers by phosphate metal affinity chromatography (PMAC) resin. The phosphoproteins were exchanged by start buffer and separated by chromatofocusing in the first dimension. Then the fractions between pH 8.5 and pH 4.0 were separated by non-porous silica (NPS) reverse-phase high performance liquid chromatography (RP-HPLC). Finally, the UV maps were converted into gel-like maps by ProteoVue software. Results Phosphoproteins of mouse liver were successfully extracted and fractionated by two dimensional liquid phase chromatographic fractionation after concentration and desalt. Then pI/UV map of mouse liver phosphoproteome was successfully set-up. There are 16 fractions between pH 8.5 and pH 4.0 after chromatofocusing in the first dimension and the UV maps of each fraction were converted into pI/UV gel-like maps. Conclusions Combination of technique of phosphoproteins enrichment and 2-D liquid phase chromatographic fractionation is an effective approach to research phosphoproteome and the key base for further identification and investigation of phosphoproteins.

The expression of microRNA (miRNA) is closely related to radio-chemosensitivity in glioma stem cells (GSCs).Moreover,the growth of glioma stem cells could be inhibited comprehensively by increasing radio-chemosensitivity and apoptosis,simultaneously with the regulation of a single miRNA,which has been confirmed by some researches.Thereby microRNA is prospective for the adoption as a specific agent in targeted therapy of glioma,so as to increase the radio-chemosensitivity in glioma stem cells.

Objective To construct the vector that expresses the fusion protein of HIV-Tat protein and red fluorescent protein(mCherry) in mammalian cells,and observe by fluorescence microscopy the intracellular transduction and localization of recombinant protein in cells,in order to obtain a useful tool for the study of the uptake mechanism and intracellular localization of HIV-TAT.Methods With the designed primer coding mCherry sequence,the mCherry gene was amplified by PCR with the vector pmCherry-C2 as template,and inserted into vector pET14b-His-TAT to construct the expression vector pET14b-His-TAT-mCherry.The constructed vector was then transformed into E.coli BL21(DE3),which had been identified by PCR and double digested with restriction endonuclease,followed by sequencing.After IPTG induction,the recombinant protein of His-TAT-mCherry was lyzed and analyzed with SDS-PAGE.Purified His-TAT-mCherry recombinant protein was added to Hela cells and the fluorescence was observed to evaluate the transduction efficiency.Results The results of identification by PCR,digestion with restriction endonuclease and sequencing indicated that the vector His-TAT-mCherry was correctly constructed.His-TAT-mCherry fusion protein was expressed in mammalian Hela cell line and purified successfully,and the fusion protein showed cellular transduction activity.It was found by fluorescence microscopy that the red fluorescence protein located mainly over the cytoplasm,and also the membrane to some extent.Conclusion The expression vector is successfully constructed for HIV-TAT labeled with mCherry sequence.Effective expression and purification of this fusion protein is achieved.It has been observed that the constructed vector may be expressed in mammalian Hela cell under active condition.Thus,it might be useful in the study of uptake mechanism and intracellular localization of HIV-TAT.

Objective: To investigate whether FasL expression by exogenous FasL gene transfer had regulatory effect on drug sensitivity of tumor cells. Methods: Exogenous Fasl, gene transfer was mediated by lipofectAMINE. The FasL expression was detected bv Immunohistochemistry and RT-PCR. The growth inhibition of cytotoxic drugs on tumor cells was measured by MTT assay. Fas mRNA expression was analyzed bv Semi-quantitative RT-PCR. Results: The eukaryotic expression vector, PcDNA3-FasL and PcDNA3 were successfully transteeted into MCF-7 cells. The transfectant cells was named as MCF-7/FL or MCF-7/Vt respectively. FasL was expressed on MCF-7/FL but not on MCF-7/Vt or MCF-7. The cytolysis ot MCF-7/FL induced by adriamycin at concentration of 0.625 ~ 2.5 ?g/ml, cisplatin of 1 .25 ~ 10 ?g/ml, or methotrexate of 3 ~ 6 ?g/ml respectively were all significantly enhanced compared with the cytolysis of MCF-7. Blocking FasL by using F(ab' )2-anti APO-1 antibody markedly reduced this up-regulated cytolysis induced by these drugs The significant up-regulation of Fas mRNA on MCF-7/FL was observed by Semi-quantitative RT-PCR after these drugs treatment. Conclusion: These findings indicated that FasL expression by gene transfer had up-regulation effect on drug sensitivity of tumor cells, this effect was associated with enhanced Fas expression induced by cytotoxic drugs.

In order to study the role of cofactor engineering in enhancing the production of S-adenosylmethionine (SAM), we altered the form and concentration of cofactor in Saccharomyces cerevisiae through gene recombination. Effects of cofactor on product synthesis, carbon and energy metabolism were analyzed aiming to provide a theoretical basis for a successful metabolic engineering of SAM producing strains. Because NADPH metabolism in mitochondrion and cytoplasm of S. cerevisiae is relatively independent, the effect of intracellular NADPH availability on the production of SAM was studied in different compartments of S. cerevisiae BY4741. The expression of NADH kinase in mitochondria (POS5 encoded) and cytoplasm (POS5Δ17 encoded) was separately confirmed using a laser scanning confocal microscope. NADPH regulation strategy enhanced SAM production. Compared with the control strain, the intracellular SAM concentration of strain NBYSM-1 was increased by 3.28 times, and the intracellular SAM concentration of strain NBYSM-2 was increased by 1.79 times at 24 h fermentation. In addition, SAM titer and NADPH/NADP⁺ ratio in strain NBYSM-1 were significantly higher than that of strain NBYSM-2. Therefore, NADPH regulation strategy will be a valuable tool for SAM production and could further improve the synthesis of a large range of cofactor-driven chemicals.

Through reviewing and summarizing classical literatures of traditional Chinese medicine (TCM) and modern medical research results,TCM kidney-marrow system was presented and initially explained.The kidney marrow system is inseparable in structure,interdependent in physiology and mutually influential in pathology.The kidney-marrow system can be applied to the treatment of major diseases in TCM,which included metabolic bone diseases,brain and neurological disorders,haematological diseases and etc.,with significant clinical effects.The in-depth study on TCM kidney marrow system will improve the clinical curative effect of major TCM clinical diseases and achieve the precision treatment in traditional medicine.It displays unique advantages of TCM holism concept and achieves new breakthrough in TCM clinical diseases.

The kidney stores essence,essence generates marrow is one of the important contents in zaag-xiang theory of traditional Chinese medicine (TCM).The kidney-marrow-brain,kidney-marrow-bone and other biological axis put forward by modern scholars took kidney-marrow system as their cores.Derived from this basis as well as under the guidance of TCM holism concept,the construction of TCM kidney-marrow system has been continuously improved;and the kidney-marrow correlation theory has been enriched.To better grasp and apply the kidney-marrow correlation theory can solve clinical problems of major diseases.However,doctors in past dynasties did not clearly define kidney-marrow system. It has not formed a complete theoretical system.Marrow in the kidney-marrow system contained brain,spinal cord and bone marrow.Marrow can generate blood,which nourishes bone together with the bone marrow.The bone marrow connects with the brain and spinal cord,which are all related to blood.Hence,the function of marrow generates blood occupied an important position in the kidney-marrow system. In this paper,the structure and function of the kidney-marrow system were preliminarily explained and constructed;and the kidney-marrow system related theories were further enriched.