ABSTRACT
BACKGROUND: Pentraxin 3 is an acute inflammatory protein of the long pentraxin subfamily. A meta-analysis was performed to assess diagnostic accuracy of pentraxin 3 for respiratory tract infections. METHODS: We identify studies examining diagnostic value of pentraxin 3 for respiratory tract infections by searching Pubmed, Web of Knowledge, and Cochrane Library. The sensitivity, specificity, negative likelihood ratio (LR), positive LR, and diagnostic odds ratio were pooled. The area under the summary receiver operator characteristic (SROC) curve and Q point value (Q*) were calculated. RESULTS: A total of 8 studies with 961 individuals were eligible for this meta-analysis. The pooled sensitivity of pentraxin 3 in diagnosis of respiratory tract infections was 0.78, the pooled specificity was 0.73, the area under the SROC curve was 0.84, and the Q* was 0.77. The area under the SROC curve of serum and bronchoalveolar lavage fluid (BALF) pentraxin 3 was 0.85 and 0.89, respectively. Meta-regression analysis revealed that cutoff value was the source of heterogeneity among the included studies. The Deek funnel plot test suggested no evidence of publication bias. Subgroup analyses showed that the area under the SROC curve of pentraxin 3 in diagnosis of ventilator-associated pneumonia (VAP) was 0.89. CONCLUSION: Pentraxin 3 has a moderate accuracy for diagnosing respiratory tract infections and VAP. The overall diagnostic value of BALF level of pentraxin 3 is superior to its serum concentration.
Subject(s)
C-Reactive Protein/analysis , Respiratory Tract Infections/blood , Respiratory Tract Infections/diagnosis , Serum Amyloid P-Component/analysis , Biomarkers , Bronchoalveolar Lavage Fluid/chemistry , Humans , Odds Ratio , Pneumonia, Ventilator-Associated/blood , Pneumonia, Ventilator-Associated/diagnosis , Predictive Value of Tests , ROC CurveSubject(s)
Forests , Life Style , Pulmonary Disease, Chronic Obstructive/therapy , Aged , China , HumansABSTRACT
In this study, two novel cationic lipids containing protonated cyclen and quaternary ammonium moieties were designed and synthesized as non-viral gene delivery vectors. The structures of the two lipids differ in their hydrophobic region (cholesterol or diosgenin). Cationic liposomes were easily prepared from the lipids individually or from the mixtures of each cationic lipid and dioleoylphosphatidylethanolamine. Several studies including DLS, gel retardation assay, and ethidium bromide intercalation assay suggest that these amphiphilic molecules are able to bind and compact DNA into nanometer particles which can be used as non-viral gene delivery agents. Our results from in vitro transfection show that in association with dioleoylphosphatidylethanolamine, two cationic lipids can induce effective gene transfection in human embryonic kidney 293 cells, although the gene transfection efficiencies of two cationic lipids were found to be lower than that of lipofectamine 2000(TM) . Besides, different cytotoxicity was found for two lipoplexes. This study demonstrates that the title cationic lipids have large potential to be efficient non-viral gene vectors.
Subject(s)
Heterocyclic Compounds/chemistry , Lipids/chemistry , Liposomes/chemistry , Quaternary Ammonium Compounds/chemistry , Cations/chemistry , Cell Survival/drug effects , Cyclams , Genetic Vectors/chemistry , Genetic Vectors/metabolism , HEK293 Cells , Humans , Lipids/chemical synthesis , Nanoparticles/chemistry , TransfectionABSTRACT
BACKGROUND: Gene therapy has tremendous potential for both inherited and acquired diseases. However, delivery problems limited their clinical application, and new gene delivery vehicles with low cytotoxicity and high transfection efficiency are greatly required. METHODS: In this report, we designed and synthesized three amphiphilic molecules (L1-L3) with the structures involving 1, 4, 7, 10-tetraazacyclododecane (cyclen), imidazolium and a hydrophobic dodecyl chain. Their interactions with plasmid DNA were studied via electrophoretic gel retardation assays, fluorescent quenching experiments, dynamic light scattering and transmission electron microscopy. The in vitro gene transfection assay and cytotoxicity assay were conducted in four cell lines. RESULTS: Results indicated that L1 and L3-formed liposomes could effectively bind to DNA to form well-shaped nanoparticles. Combining with neutral lipid DOPE, L3 was found with high efficiency in gene transfer in three tumor cell lines including A549, HepG2 and H460. The optimized gene transfection efficacy of L3 was nearly 5.5 times more efficient than that of the popular commercially available gene delivery agent Lipofectamine 2000™ in human lung carcinoma cells A549. In addition, since L1 and L3 had nearly no gene transfection performance in normal cells HEK293, these cationic lipids showed tumor cell-targeting property to a certain extent. No significant cytotoxicity was found for the lipoplexes formed by L1-L3, and their cytotoxicities were similar to or slightly lower than the lipoplexes prepared from Lipofectamine 2000™. CONCLUSION: Novel cyclen-based cationic lipids for effective in vitro gene transfection were founded, and these studies here may extend the application areas of macrocyclic polyamines, especially for cyclen.
Subject(s)
Gene Transfer Techniques , Heterocyclic Compounds/chemistry , Lipids/chemistry , Neoplasms/metabolism , Cations , Cell Death , Cell Line, Tumor , Cyclams , DNA/metabolism , Electrophoretic Mobility Shift Assay , Ethidium/metabolism , Humans , Light , Liposomes/chemistry , Liposomes/ultrastructure , Microscopy, Fluorescence , Particle Size , Phosphatidylethanolamines/chemistry , Scattering, Radiation , Static Electricity , TransfectionABSTRACT
In this report, as candidates for non-viral gene vectors, cationic lipids L1, L2 and L3 based on protonated cyclen and different hydrophobic groups (cholesterol, dodecanol or diosgenin) linked by PNA monomer were designed and synthesized. Their liposomes were easily prepared by mixing the synthesized lipids with dioleoylphosphatidyl ethanolamine (DOPE) under appropriate mole ratios. Agarose gel retardation and fluorescent titration by ethidium bromide (EB) showed the strong DNA-binding ability with the K(sv) values of 1.21 × 10(7), 3.76 × 10(6) and 2.90 × 10(6) M(-1) for the liposomes formed from L1-L3, respectively. These liposomes could retard pDNA at an N/P ratio of 3 and form lipoplexes with sizes around 200-300 nm and zeta-potential values of +20-50 mV at N/P ratio from 4 to 10. Besides, the cytotoxicity of the three lipoplexes assayed by MTT is quite different. The results from in vitro transfection in HEK 293T and A549 cell lines showed that the transfection efficiency of L3/DOPE/DNA lipoplex at an N/P ratio of 6 and lipid/DOPE mole ratio of 1:2 is slightly higher than that of Lipofectamine 2000™, indicating that the title PNA monomer-based cationic lipids have great potential to be efficient non-viral gene vector.
Subject(s)
Heterocyclic Compounds/chemistry , Lipids/chemistry , Peptide Nucleic Acids/chemistry , Transfection , Uracil/chemistry , Cations , Cyclams , Magnetic Resonance Spectroscopy , Spectrometry, Mass, Electrospray IonizationABSTRACT
In this study, two novel protonated cyclen and imidazolium salt-containing cationic lipids differing only in their hydrophobic region (cholesterol or diosgenin) have been designed and synthesized for gene delivery. Cationic liposomes were easily prepared from each of these lipids individually or from the mixtures of each cationic lipid and dioleoylphosphatidyl ethanolamine (DOPE). Several studies including DLS, gel retardation assay, ethidium bromide intercalation assay, and TEM demonstrated that these amphiphilic molecules are able to bind and compact DNA into nanometer particles that could be used as non-viral gene delivery agents. Our results from in vitro transfection showed that in association with DOPE, two cationic lipids can induce effective gene transfection in HEK293 cells. Furthermore, the gene transfection efficiencies of two cationic lipids were dramatically increased in the presence of calcium ion (Ca(2+)). It is notable that the gene transfection abilities of two cationic lipids were maintained in the presence of 10% serum. Besides, different cytotoxicity was found for two lipoplexes. This study demonstrates that the title cationic lipids have large potential to be efficient non-viral gene vector.
Subject(s)
Drug Delivery Systems , Genetic Therapy/methods , Heterocyclic Compounds/chemistry , Imidazoles/chemical synthesis , Lipids/chemical synthesis , Liposomes/chemistry , Cations/chemical synthesis , Cyclams , DNA/chemistry , Drug Compounding , Genetic Vectors , HEK293 Cells , Humans , Lipids/chemistry , Nanoparticles , TransfectionABSTRACT
The synthesis and characterization of a novel macrocyclic polyamine cationic lipid containing an imidazolium salt group is reported. Its interaction with plasmid DNA was studied by gel electrophoresis and fluorescence quenching experiments. The transfection activity of target compound as a gene delivery vector was also investigated. The results showed that the synthesized macrocyclic polyamine cationic lipid has high binding and condensation ability of DNA under physiological conditions probably because of the cooperation effect of macrocyclic polyamine (Cyclen) and an imidazolium salt group. This novel lipid could transfer plasmid DNA into cell in in vitro experiment without the use of any extraneous agent.
