ABSTRACT
Two hitherto unknown iboga-type indole alkaloids, namely (3R)-7,19-di-epi-3-methoxytabernoxidine (1) and (3R,19R)-19-hydroxy-3-(2-oxopropyl)voacangine (2), together with eight known alkaloids (3-10), were isolated from the twigs and leaves of Tabernaemontana divaricata. Their structures were established on the basis of spectroscopic data interpretation, single crystal X-ray diffraction analysis and circular dichroism spectrum.
ABSTRACT
OBJECTIVE: To investigate the Raman spectral characteristics of leukemia cells from 4 patients with acute promyelocytic leukemia (M3) and 3 patients with acute monoblastic leukemia (M5), establish a novel Raman label-free method to distinguish 2 kinds of acute myeloid leukemia cells so as to provide basis for clinical research. METHODS: Leukemia cells were collected from bone marrow of above-mentioned patients. Raman spectra were acquired by Horiba Xplora Raman spectrometer and Raman spectra of 30-50 cells from each patient were recorded. The diagnostic model was established according to principle component analysis (PCA), discriminant function analysis (DFA) and cluster analysis, and the spectra of leukemia cells from 7 patients were analyzed and classified. Characteristics of Raman spectra were analyzed combining with ultrastructure of leukemia cells. RESULTS: There were significant differences between Raman spectra of 2 kinds of leukemia cells. Compared with acute monoblastic leukemia cells, the spectra of acute promyelocytic leukemia cells showed stronger peaks in 622, 643, 757, 852, 1003, 1033, 1117, 1157, 1173, 1208, 1340, 1551, 1581 cm-1. The diagnostic models established by PCA-DFA and cluster analysis could successfully classify these Raman spectra of different samples with a high accuracy of 100% (233/233). The model was evaluated by "Leave-one-out" cross-validation and reached a high accuracy of 97% (226/233). CONCLUSION: The level of macromolecules of M3 cells is higher than that of M5. The diagnostic models established by PCA-DFA can classify these Raman spectra of different cells with a high accuracy. Raman spectra shows consistent result with ultrastructure by TEM.
Subject(s)
Leukemia, Myeloid, Acute , Cluster Analysis , Humans , Leukemia, Monocytic, Acute , Principal Component Analysis , Spectrum Analysis, RamanABSTRACT
Cardiotoxicity is one of the most serious side effects of new drugs. Early detection of the drug induced cardiotoxicity based on the biomarkers provides an important preventative strategy for detecting potential cardiotoxicity of candidate drugs. In this study, we aim to identify the predictive genomics biomarkers for drug-induced cardiac toxicity based on the RTCA coupled with PCR Array technology in primary cells. Three prototypical cardiotoxic compounds (doxorubicin, isoproterenol, ouabain) with different mechanisms were firstly real-time monitored to diagnose the cytotoxicity by using the RTCA, while the functional alterations of cardiomyocytes were also monitored by analyzing the beating frequency of cardiomyocytes. Then cardiac specific toxicity gene expression changes were studied by using the technology of PCR Array, which can detect the changes of 84 cardiac functions related genes. Rps6kb1 was identified to be the common cardiac biomarkers by using multivariate statistical and integration analyses. The biomarker was further verified by selecting other drugs with or without cardiotoxicity, and the results showed that the gene exhibited specific changes in cardiac toxicity. Moreover, IPA was applied to combine relevant pathways of Rps6kb1, and identify the main types of cardiac toxicity. These results would further enrich the evaluating strategy of drug-induced cardiotoxicity in vitro, and Rps6kb1 could be used as the specific biomarker of cardiotoxcity during safety assessment of the novel drug candidates.
Subject(s)
Doxorubicin/toxicity , Gene Expression Profiling/methods , Heart Diseases/chemically induced , Heart Diseases/genetics , Isoproterenol/toxicity , Myocytes, Cardiac/drug effects , Oligonucleotide Array Sequence Analysis , Ouabain/toxicity , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Toxicity Tests , Animals , Animals, Newborn , Cardiotoxicity , Cells, Cultured , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Genetic Markers , Heart Diseases/metabolism , Heart Diseases/physiopathology , Heart Rate/drug effects , Myocytes, Cardiac/metabolism , Rats, Sprague-Dawley , Reproducibility of Results , Ribosomal Protein S6 Kinases, 70-kDa/genetics , Ribosomal Protein S6 Kinases, 70-kDa/metabolismABSTRACT
OBJECTIVE: To analyze and evaluate the application of spinning disk confocal microscopy and imaging analysis software in movement and phagocytosis of neutrophils. METHODS: Neutrophils were isolated from bone marrow by centrifugation on discontinuous Percoll gradient, and then were stained with PE Gr-1 antibody and mixed with FITC-labeled Zymosan A bioparticles. Multichannel time-lapse videos were captured by using the spinning disk confocal microscopy. The result was analyzed by using volocity and ImageJ software, the parameters associated with movement and phagocytosis of neutrophils were analyzed, including morphological changes, cell tracking, pseudopod dynamics, binding and phagocytosis index. RESULTS: Most neutrophils would be polarized in response to Zymosan particles during a short time. Binding and phagocytosis process occured in forty minutes. CONCLUSION: A method of precisely quantifying the movement and phagocytosis of neutrophils using microscopic imaging and imaging analysis technique has been set up successfully. Using this method, biological activity and function of neutrophils can be evaluated visually and rapidly. The physiologically rapid response to Zymosan particles can be applied to the neutrophils function research in the future.
Subject(s)
Cell Movement , Neutrophils , Phagocytosis , Antibodies , Bone Marrow , Humans , Microscopy , ZymosanABSTRACT
Six new ent-pimarane diterpenoids, namely, ent-14beta,16-epoxy-8-pimarene-3beta,15alpha-diol (1), 7beta-hydroxydarutigenol (2), 9beta-hydroxydarutigenol (3), 16-O-acetyldarutigenol (4), 15,16-di-O-acetyldarutoside (5), and 16-O-acetyldarutoside (6), were isolated from the ethanol extract of Siegesbeckia orientalis. Their structures were elucidated on the basis of spectroscopic studies, and the absolute configuration of 1 was established by the modified Mosher's method. Furthermore, the structure of ent-12beta,16-epoxy-8-pimarene-2alpha,15beta,19-triol, recently isolated from the same plant, should be revised as ent-14beta,16-epoxy-8-pimarene-2alpha,15alpha,19-triol (10).
Subject(s)
Abietanes/chemistry , Abietanes/isolation & purification , Asteraceae/chemistry , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Humans , Molecular Structure , Nuclear Magnetic Resonance, BiomolecularABSTRACT
Two new guaiane-type sesquiterpene lactones with an unusual sulfonic acid group, sulfocostunolide A (1) and sulfocostunolide B (2), were isolated from the roots of Saussurea lappa. Their structures were elucidated on the basis of extensive spectroscopic analysis.
