ABSTRACT
Objective To develop a sheet for postural change during painless colonoscopy.Methods The sheet was made of medical canvas whose body had a size of 110 cm (length) × 80 (width) cm.The body had a waistband at its each side,and the waistband had 5 nylon Velcro at each side to tie it to the patient's waist.There were 6 canvas handles at the back of the body,which were in parallel with the waistband for drawing the patient during postural change.A tearing mechanism with Velcro existed at the hip area to facilitate the hip exposure during colonoscopy.Two shoes covers were at right under the body to hold the feet when the patient bent his knees.Results The sheet significantly shortened operation time,increased working efficiency and protected the patient's privacy when compared with manual operation.Conclusion The sheet gains high safety,convenience and efficiency for postural change during painless colonoscopy,and thus is worthy promoting in the hospitals.
ABSTRACT
<p><b>OBJECTIVE</b>To construct an cell penetrating peptide-based expression vector capable of targeted delivery of proteins into the cell nuclei and study its function of protein transduction.</p><p><b>METHODS</b>The fusion protein expression vector pET14b-HC(L)NE (pET14-b-His-CPP-Linker-NLS-EGFP) incorporating cell penetrating peptide (CPP), nuclear localization signal(NLS), linker and enhanced green fluorescent protein (EGFP) was constructed based on His-tagged pET14b-HE (pET14b-His-EGFP) by site-directed mutagenesis PCR method. After identification by enzyme digestion and DNA sequencing, the recombinant plasmid was transformed into BL21(DE(3)) strain. The HC(L)NE fusion protein was expressed following IPTG induction and purified with Ni(2+)-NTA affinity chromatography. After dialysis and filtration, the HC(L)NE fusion protein was added into cultured eukaryotic cells. The protein transduction in the living cells was observed under fluorescence microscope and analyzed by Western blotting.</p><p><b>RESULTS</b>Enzyme digestion and DNA sequencing confirmed successful construction of the pET14b-HC(L)NE vector, and the fusion protein efficiently expressed in E. coli. Protein transduction experiments in eukaryotic cells revealed that the fusion protein could rapidly penetrate the cell membrane and reach the cell nucleus, and this internalization was time- and concentration-dependent.</p><p><b>CONCLUSION</b>The cell penetrating peptide-based expression vector for targeted protein delivery to the cell nucleus has been successfully constructed, and a transport system that can delivery exogenous proteins or polypeptides into the cytoplasm and cell nucleus is established, which provides an economical and efficient means for functional study of the proteins and polypeptide in cells and targeted drug delivery.</p>
Subject(s)
Animals , Humans , Mice , Blotting, Western , COS Cells , Cell Line , Cell Membrane , Metabolism , Cell Nucleus , Metabolism , Chlorocebus aethiops , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , HeLa Cells , Microscopy, Fluorescence , NIH 3T3 Cells , Nuclear Localization Signals , Genetics , Peptides , Genetics , Metabolism , Protein Transport , Recombinant Fusion Proteins , Genetics , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To explore the reasons for the low intracellular transduction efficiency of a previously constructed His-TAT-Flag recombinant protein and establish a more efficient transduction system.</p><p><b>METHODS</b>The Flag tag of pET14b-His- Tat-Flag vector was deleted with PCR mutant kit, and enhanced green fluorescent protein (EGFP) coding sequence was inserted into the new pET14b-His-TAT recombinant vector. Enzyme digestion and DNA sequencing were performed for identification of pET14b-His-TAT-EGFP vector, which was then transformed into E. coli BL21(DE(3)). After IPTG induction, the recombinant protein of His-TAT-EGFP was isolated and analyzed with SDS-PAGE. Purified His-TAT-EGFP recombinant protein was added to ECV304 cells and the fluorescence was observed to evaluate the transduction efficiency.</p><p><b>RESULTS</b>pET14b-His-TAT vector and pET14b-His-TAT- EGFP vector were successfully constructed, which was identified with enzyme digestion and DNA sequencing. His-TAT-EGFP fusion protein was expressed and purified successfully and showed cellular transduction activity.</p><p><b>CONCLUSION</b>The prokaryotic expression vector has been successfully constructed by modifying pET14b-His-TAT-Flag, and the expressed and purified recombinant protein of His-TAT-EGFP possesses high efficiency of intracellular transduction activity.</p>
Subject(s)
Humans , Amino Acid Sequence , Cell Line , Electrophoresis, Polyacrylamide Gel , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , HIV-1 , Genetics , Microscopy, Fluorescence , Molecular Sequence Data , Protein Transport , Recombinant Fusion Proteins , Genetics , Metabolism , Transfection , Methods , tat Gene Products, Human Immunodeficiency Virus , Genetics , MetabolismABSTRACT
<p><b>AIM</b>To investigate the bioactive constituents from the mycelium of Penicillium thomii. Which isolated from Anemone collected in Qingdao beach.</p><p><b>METHODS</b>The constituents were separated by using various chromatography and the structures were identified on the basis of extensive spectral analysis.</p><p><b>RESULTS</b>Five compounds, namely penicillixanthone A (I), p-methylbenzolic acid (II), 1-O-hexadecanoyl-2-O-(9-octadecenoyl)-3-O-(9, 12-octadecadienoyl) glycerol (III), 5 alpha, 8 alpha-epidioxy-24 zeta-methylcholesta-6, 22-dien-3 beta-ol (IV) and 1, 6, 8-trihydroxyl-3-methyl-9, 10-anthracenedione (V), were isolated from the mycelium of Penicillium thomii.</p><p><b>CONCLUSION</b>Penicillixanthone A is a new compound, while the others are isolated from Penicillium thomii for the first time.</p>
Subject(s)
Animals , Molecular Conformation , Molecular Structure , Penicillium , Chemistry , Sea Anemones , Microbiology , Xanthones , ChemistryABSTRACT
Objective To investigate the roles of adrenomedullin(ADM) and endothelin-1(ET-1) in left-to-right shunt congenital heart disease(CHD) complicated with congestive heart failure(CHF).Methods Plasma ADM and ET-1 concentrations were measured by radioimmunoassay in heart failure stage and recovery period of 18 infants with left-to-right shunt CHD.Also 20 healthy infants acted as control group were measured.Results Plasma concentrations of ADM and EF-1 were significantly higher in heart failure stage of left-to-right CHD infants than those in their recovery period,and both in their heart failure stage and in their recovery period were higher than that in control group(P