ABSTRACT
Bacteriophage lysin is characterized by high stability, wide bactericidal activity and efficacy, and safty. It is able to lyse bacteria specifically and is not susceptible to bacterial resistance. In the presence of phage, phage infecting bacteria and coding for endolysin then it lyse the bacteria, In the absence of the phage, holin assists the endolysin lysis the bacteria from external. In addition, the research progress on the treatment of Gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus, Streptococcus pyogenes, and Gram-negative bacteria, such as Acinetobacter baumannii and Pseudomonas aeruginosa, was also discussed in this paper.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of latent and active pulmonary tuberculosis(TB) on expression of miR-29 family and target gene IFN-γ in CD4(+)T cells.</p><p><b>METHODS</b>Subjects from two hospitals of Weifang were enrolled from March 2012 to December 2012 and divided into three groups: active TB group(30 cases), latent tuberculosis infection(LTBI) group(25 cases) and healthy control group(30 cases). CD4(+) T cells in blood were collected from the three groups.Levels of miR-29a, miR-29b and miR-29c were measured by nucleic acid hybridization and RT-qPCR.Expression of IFN-γ was analyzed by RT-qPCR. Target genes of miR-29 family were predicted with both TargetScan and PicTar.GO annotation and pathway overrepresentation were further analyzed with David database and Cytoscape.</p><p><b>RESULTS</b>Levels of miR-29a, miR-29b and miR-29c showed significant differences among the three groups(P < 0.05): levels of miR-29b and miR-29c in the active TB group(561.63 ± 65.36, 281.85 ± 42.78) were higher than the healthy controls(260.74 ± 38.69, 128.21 ± 19.98), but lower than the LTBI group(2030.29 ± 321.68, 620.93 ± 79.14); expression of miR-29a in the healthy control group(913.95 ± 104.73) were higher than the active TB group(323.37 ± 54.38), but lower than the LTBI group(4782.13 ± 567.81).Level of IFN-γ showed significant differences among the three groups(P < 0.05): level of IFN-γ in the LTBI group(0.45 ± 0.09) were lower than the healthy controls(1.00), but higher than the active TB group(0.11 ± 0.03). The target genes of miR-29 family mainly existed in molecular function such as extracellular matrix structural constituent and transcription regulator activity.In KEGG pathway, the gene set mostly existed in signaling pathway such as Focal adhesion,ECM-receptor interaction and mTOR signaling pathway.</p><p><b>CONCLUSION</b>The expression of miR-29 family was increased and target gene IFN-γ in CD4(+) T cells was decreased by latent and active pulmonary TB, which might play important role in alteration of signal pathway.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , CD4-Positive T-Lymphocytes , Metabolism , Case-Control Studies , Computational Biology , Gene Expression , Interferon-gamma , Genetics , Metabolism , Latent Tuberculosis , Genetics , Allergy and Immunology , MicroRNAs , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To construct a plasmid carrying granulysin (GLS) and to study the effect of the GLS on apoptosis, mitochondrial transmembrane potential and cytochrome C release of SMMC-7721 cells.</p><p><b>METHODS</b>The coding sequence of the GLS was amplified from the total RNA of human CTL cells, and it was inserted into pBudCE4.1 plasmid and then it was used to transfect SMMC-7721 cells. The expression of GLS was detected by RT-PCR and confirmed by immunocytochemistry method. Cell apoptosis was ascertained by Hoechst staining and electron microscopy; mitochondrial transmembrane potential was detected using Mitocapture and cytochrome C release was studied using Western blot.</p><p><b>RESULTS</b>Recombinant pBudCE4.1/GLS plasmid was successfully constructed. GLS protein was successfully expressed in the SMMC-7721 cells and it induced apoptosis of the SMMC-7721 cells, and at the same time, mitochondrial transmembrane potential was reduced and cytochrome C was released from mitochondria into the cytosol.</p><p><b>CONCLUSIONS</b>GLS gene carried by recombinant plasmid could express in SMMC-7721 cells and induce cells apoptosis. The change of mitochondrial transmembrane potential and the release of cytochrome C might be one of the key factors of apoptosis induced by GLS.</p>
Subject(s)
Humans , Antigens, Differentiation, T-Lymphocyte , Pharmacology , Apoptosis , Cell Line, Tumor , Cytochromes c , Metabolism , Membrane Potential, Mitochondrial , Mitochondria , Metabolism , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the differentially expressed mitochondrial proteins in hydroxycamptothecin (HCPT)-treated SMMC-7721 cells by comparative proteomic analysis.</p><p><b>METHODS</b>Apoptosis of SMMC-7721 cells were induced by using HCPT and their mitochondria were isolated with a mitochondria isolation kit for cultured cells. Three different solubility protein fractions were extracted with ReadyPrep Sequential Extraction Kit and were separated by two-dimensional gel electrophoresis (2-DE). PDQuest software was used to differentiate mitochondrial proteins between control cells and HCPT-treated cells. Matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) was used to identify some of the different proteins.</p><p><b>RESULTS</b>Highly purified mitochondria and high resolution 2-DE patterns of the proteins were obtained. Forty-four mitochondrial protein spots from the HCPT-treated cells showed different expressions compared to those of the control cells. Twenty of the different protein spots were analyzed by MALDI-TOF-MS.</p><p><b>CONCLUSION</b>Differently expressed mitochondrial proteins in HCPT-treated cells and control cells were obtained in this study. This will be of help to understand the mechanism by which HCPT induces cell apoptosis.</p>