ABSTRACT
A Gram-negative, aerobic, nonmotile, rod-shaped and yellow-pigment-producing bacteria was isolated from Baima snow mountain of Diqing Tibetan Autonomous Prefecture in Yunnan province, south-west China and characterized using a polyphasic approach. The results of 16S rRNA gene sequence similarity analysis showed that strain YIM B04101T was closely related to the type strain of Dyadobacter koreensis DSM 19938T (97.81%) and Dyadobacter frigoris AR-3-8T (97.95%). The predominant respiratory quinone was menaquinone-7 (MK-7). The major polar lipid was phosphatidylethanolamine. The major fatty acids were summed feature 3 (C16:1ω7c/C16:1ω6c), C18:1ω9c and C16:0. The DNA G + C content was 43.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain YIM B04101T belonged to a cluster comprising species of the genus Dyadobacter. However, it differed from its closest relative, Dyadobacter koreensis KCTC 12537T and Dyadobacter frigoris AR-3-8T, in many physiological properties. Based on these phenotypic characteristics and phylogenetic distinctiveness, strain YIM B04101T is considered to be a novel species of the genus Dyadobacter, for which the name Dyadobacter diqingensis sp. nov. is proposed. The type strain is YIM B04101T (= CGMCC 1.19249T = CCTCC AB 2021270).
Subject(s)
Fatty Acids , Snow , Bacterial Typing Techniques , China , Cytophagaceae , DNA, Bacterial/genetics , Fatty Acids/analysis , Phospholipids , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , TibetABSTRACT
BACKGROUND: The expression profiles and molecular mechanisms of CXC chemokine receptors (CXCRs) in Lung adenocarcinoma (LUAD) have been extensively explored. However, the comprehensive prognostic values of CXCR members in LUAD have not yet been clearly identified. METHODS: Multiple available datasets, including Oncomine datasets, the cancer genome atlas (TCGA), HPA platform, GeneMANIA platform, DAVID platform and the tumor immune estimation resource (TIMER) were used to detect the expression of CXCRs in LUAD, as well as elucidate the significance and value of novel CXCRs-associated genes and signaling pathways in LUAD. RESULTS: The mRNA and/or protein expression of CXCR1, CXCR2, CXCR3, CXCR4, CXCR5 and CXCR6 displayed predominantly decreased in LUAD tissues as compared to normal tissues. On the contrary, compared with the normal tissues, the expression of CXCR7 was significantly increased in LUAD tissues. Subsequently, we constructed a network including CXCR family members and their 20 related genes, and the related GO functions assay showed that CXCRs connected with these genes participated in the process of LUAD through several signal pathways including Chemokine signaling pathway, Cytokine-cytokine receptor interaction and Neuroactive ligand-receptor interaction. TCGA and Timer platform revealed that the mRNA expression of CXCR family members was significantly related to individual cancer stages, cancer subtypes, patient's gender and the immune infiltration level. Finally, survival analysis showed that low mRNA expression levels of CXCR2 (HR = 0.661, and Log-rank P = 1.90e-02), CXCR3 (HR = 0.674, and Log-rank P = 1.00e-02), CXCR4 (HR = 0.65, and Log-rank P = 5.01e-03), CXCR5 (HR = 0.608, and Log-rank P = 4.80e-03) and CXCR6 (HR = 0.622, and Log-rank P = 1.85e-03) were significantly associated with shorter overall survival (OS), whereas high CXCR7 mRNA expression (HR = 1.604, and Log-rank P = 4.27e-03) was extremely related with shorter OS in patients. CONCLUSION: Our findings from public databases provided a unique insight into expression characteristics and prognostic values of CXCR members in LUAD, which would be benefit for the understanding of pathogenesis, diagnosis, prognosis prediction and targeted treatment in LUAD.
Subject(s)
Adenocarcinoma of Lung , Adenocarcinoma , Lung Neoplasms , Adenocarcinoma/pathology , Adenocarcinoma of Lung/genetics , Adenocarcinoma of Lung/pathology , Humans , Lung Neoplasms/pathology , Prognosis , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
OBJECTIVE: To summarize the clinical characteristics of adult cases of paragonimiasis with lung masses as the main manifestation in Xishuangbanna, Yunnan Province, analyze the causes of misdiagnosis, and improve the levels of clinical diagnosis and treatment. METHOD: We conducted a retrospective analysis of the clinical data and diagnosis and treatment of 8 adult cases of paragonimiasis with lung masses as the main manifestation that were diagnosed in the Oncology Department of People's hospital of Xishuangbanna Dai Autonomous Prefecture from July 2014 to July 2019. RESULT: All 8 patients were from epidemic paragonimiasis areas and had a confirmed history of consuming uncooked freshwater crabs. The clinical manifestations were mainly fever, dry cough, and chest pain. The disease durations were long, and peripheral blood eosinophil counts were elevated. The cases had been misdiagnosed as pneumonia or pulmonary tuberculosis. After years of anti-inflammatory or anti-tuberculosis treatment, the symptoms had not improved significantly. Patients eventually sought treatment from the oncology department for hemoptysis. Chest computed tomography showed patchy consolidation in the lungs, with nodules, lung masses, and enlarged mediastinal lymph nodes. CONCLUSION: Paragonimiasis is a food-borne parasitic disease. Early clinical manifestations and auxiliary examination results are nonspecific. The parasite most often invades the lungs, and the resulting disease is often misdiagnosed as pneumonia, pulmonary tuberculosis, or lung cancer (Acta Trop 199: 05074, 2019). To avoid misdiagnosis, clinicians should inquire, in detail, about residence history and history of unclean food and exposure to infected water and make an early diagnosis based on the inquired information and imaging examination results. For patients who have been diagnosed with pneumonia or pulmonary tuberculosis and whose symptoms do not improve significantly after anti-inflammatory or anti-tuberculosis treatments, their epidemiological history should be traced to further conduct differential diagnosis and avoid misdiagnosis.
Subject(s)
Lung Diseases, Parasitic/diagnosis , Lung/diagnostic imaging , Paragonimiasis/diagnosis , Animals , Antibodies, Helminth/analysis , China/epidemiology , DNA, Helminth/analysis , Diagnosis, Differential , Diagnostic Errors , Enzyme-Linked Immunosorbent Assay , Female , Humans , Incidence , Lung/parasitology , Lung Diseases, Parasitic/epidemiology , Lung Diseases, Parasitic/parasitology , Male , Middle Aged , Paragonimiasis/drug therapy , Paragonimiasis/parasitology , Paragonimus/genetics , Paragonimus/immunology , Retrospective Studies , Thorax/pathology , Tomography, X-Ray ComputedABSTRACT
OBJECTIVE: To compare the efficacies of 0.02% atropine eye drops and orthokeratology to control axial length (AL) elongation in children with myopia. METHODS: In this historical control study, 247 children with myopia whose administration of 0.02% atropine (n=142) or underwent orthokeratology from an earlier study (n=105, control group) were enrolled. Data on AL and other baseline parameters were recorded at baseline and after 1 and 2 years of treatment. RESULTS: The mean changes in AL in the first and second years of treatment were 0.30±0.21 and 0.28±0.20 mm, respectively, in the 0.02% atropine group and 0.16±0.20 and 0.20±0.16 mm, respectively, in the orthokeratology group. Axial length elongations after 2 years of treatment were 0.58±0.35 and 0.36±0.30 mm (P=0.007) in the 0.02% atropine and orthokeratology groups, respectively. Multivariate regression analyses showed that the AL elongation was significantly faster in the 0.02% atropine group than in the orthokeratology group (ß=0.18, P=0.009). In multivariate regression analyses, younger age and shorter baseline AL were associated with a rapid AL elongation in the 0.02% atropine group (ßage=-0.04, P=0.01; ßAL=-0.17, P=0.03), while younger age, lower baseline spherical equivalent refractive error (SER), and shorter baseline AL were associated with a greater increase in AL in the orthokeratology group (ßage=-0.03, P=0.04; ßSER=0.06, P=0.03; ßAL=-0.11, P=0.009). Faster AL elongation was found in the 0.02% atropine group compared with the orthokeratology group at higher baseline SER (P=0.04, interaction test). CONCLUSION: Within the limits of this study design, orthokeratology seems to be a better method for controlling AL elongation compared with administration of 0.02% atropine in children with higher myopia over a treatment period of 2 years.
Subject(s)
Myopia , Orthokeratologic Procedures , Atropine , Axial Length, Eye , Child , Humans , Infant, Newborn , Myopia/therapy , Refraction, OcularABSTRACT
PURPOSE: To investigate the effect of orthokeratology (OK) lens on axial length (AL) elongation in unilateral myopia and bilateral myopia with anisometropia children. METHODS: Twenty-seven unilateral myopia (group 1) and 25 bilateral myopia with anisometropia children (group 2) were involved in this 1-year retrospective study. The eyes with higher spherical equivalent refractive error (SER) were assigned to the H eyes subgroup and the fellow eyes with lower SER to the L eyes subgroup in the two groups. RESULTS: The mean change in AL of H eyes and L eyes were 0.11 ± 0.19 mm, 0.30 ± 0.28 mm in group 1 (P = 0.04) and 0.09± 0.14mm, 0.13± 0.16mm in group 2 (P = 0.36), respectively. Multivariate regression analyses showed that significant difference of change in AL was found between H eyes and L eyes in group1 (ß=0.25, P = 0.03), but no difference in group 2 (ß=0.09, P = 0.12). The AL of H eyes in group 1 and group 2, H eyes in group 1 and L eyes in group 2 had the same increased rate (ß= -0.04, P = 0.43; ß = 0.02, P = 0.56). CONCLUSIONS: Monocular OK lens is effective on suppression AL elongation of the myopic eyes and reduce anisometropia value in unilateral myopic children. The OK lens can control the AL elongation in both eyes at the same rate, but it cannot reduce anisometropia value in bilateral myopia with anisometropia children after 1-year follow-up.
Subject(s)
Anisometropia/therapy , Axial Length, Eye/pathology , Contact Lenses , Myopia/therapy , Orthokeratologic Procedures , Adolescent , Anisometropia/physiopathology , Child , Corneal Topography , Female , Humans , Male , Myopia/physiopathology , Refraction, Ocular , Retrospective StudiesABSTRACT
CXCL3 belongs to the CXC-type chemokine family and is known to play a multifaceted role in various human malignancies. While its clinical significance and mechanisms of action in uterine cervical cancer (UCC) remain unclear. This investigation demonstrated that the UCC cell line HeLa expressed CXCL3, and strong expression of CXCL3 was detected in UCC tissues relative to nontumor tissues. In addition, CXCL3 expression was strongly correlated with CXCL5 expression in UCC tissues. In vitro, HeLa cells overexpressing CXCL3, HeLa cells treated with exogenous CXCL3 or treated with conditioned medium from WPMY cells overexpressing CXCL3, exhibited enhanced proliferation and migration activities. In agreement with these findings, CXCL3 overexpression was also associated with the generation of HeLa cell tumor xenografts in athymic nude mice. Subsequent mechanistic studies demonstrated that CXCL3 overexpressing influenced the expression of extracellular signal-regulated kinase (ERK) signaling pathway associated genes, including ERK1/2, Bcl-2, and Bax, whereas the CXCL3-induced proliferation and migration effects were attenuated by exogenous administration of the ERK1/2 blocker PD98059. The data of the current investigation support that CXCL3 appears to hold promise as a potential tumor marker and interference target for UCC.
Subject(s)
Chemokines, CXC/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Uterine Cervical Neoplasms/enzymology , Adult , Aged , Animals , Apoptosis , Cell Movement , Cell Proliferation , Chemokine CXCL5/genetics , Chemokine CXCL5/metabolism , Chemokines, CXC/genetics , Female , Gene Expression Regulation, Neoplastic , HeLa Cells , Humans , Mice, Nude , Middle Aged , Neoplasm Invasiveness , Paracrine Communication , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Signal Transduction , Up-Regulation , Uterine Cervical Neoplasms/genetics , Uterine Cervical Neoplasms/pathology , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolismABSTRACT
Human corneal fibroblasts (HCFs) are implicated in corneal neovascularization (CRNV). The mechanisms underlying the inflammatory response in HCFs and the development of CRNV were explored in this study. Alkali burns were applied to the corneas of rats to establish a CRNV model. The expression of long noncoding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) and mRNA and protein levels of nuclear factor kappa B (NF-κB)- activating protein (NKAP) were examined by quantitative real-time (qRT-PCR) and Western blot methods, respectively. Lipopolysaccharide (LPS) is used to stimulate HCFs for inflammatory response. The level of inflammation factors in HCF supernatant was detected using an enzyme-linked immunosorbent assay (ELISA). Binding and interactions between NEAT1 and miRNA 1246 (miR-1246) were determined by RNA immunoprecipitation (RIP) and RNA pull-down assays in HCFs. Compared with the control group (n = 6), NEAT1 was upregulated in the corneas of the CRNV rat model (n = 6). The expression of NEAT1 in HCFs was upregulated by LPS. Downregulation of NEAT1 suppressed the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). NEAT1 could bind and interact with miR-1246. LPS regulated the expression of NKAP and NF-κB signaling via the NEAT1/miR-1246 pathway. Downregulation of NEAT1 in vivo inhibited CRNV progression in the CRNV rat model. The lncRNA NEAT1 induced secretion of inflammatory factors, mediated by NF-κB, by targeting miR-1246, thereby promoting CRNV progression.
Subject(s)
Corneal Neovascularization/metabolism , Inflammation/metabolism , RNA, Long Noncoding/metabolism , Animals , Blotting, Western , Corneal Neovascularization/genetics , Enzyme-Linked Immunosorbent Assay , Immunoprecipitation , Inflammation/genetics , Male , RNA, Long Noncoding/genetics , Rats , Real-Time Polymerase Chain ReactionABSTRACT
There is increasing evidence reporting that as a common phenomenon in MetS relative diseases, insulin resistance (IR) is regarded as an independent etiological factor and a warning indicator of MetS occurrence. Therefore, for the special group (overweight or obesity), clinical regular monitoring of IR is an important basis for the prevention and early intervention of MetS relative diseases. This surveys reveals that human umbilical cord mesenchymal stem cells (HUC-MSCs)possess a kind of potential: it may become a possible theraphy for IR in type 2 diabetes mellitus (T2DM) and related diseases. Specific emphasis is focused on evaluating the improvement IR function of HUC-MSCs under the background of development in vitro and in vivo. Next, the action mechanisms of HUC-MSCs is discussed, and some of their advantages and disadvantages in the course of clinic application are presented. The final section highlights the application of HUC-MSCs in T2DM and relative diseases at this stage. Up to now, although many questions remain unresolved, we still consider that HUC-MSCs is one of the best therapy ameliorating IR in the future.
Subject(s)
Insulin Resistance , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Umbilical Cord/cytology , Diabetes Mellitus/therapy , Humans , Translational Research, BiomedicalABSTRACT
INTRODUCTION: We have previously indicated that CXCL3 was upregulated in the tissues of prostate cancer, and exogenous administration of CXCL3 played a predominant role in the tumorigenicity of prostate cancer cells. In the present study, we further explored the role and the underlying mechanism of CXCL3 overexpression in the oncogenic potential of prostate cancer in an autocrine/paracrine fashion. METHODS: CXCL3-overexpressing prostate cancer cell line PC-3 and immortalized prostate stromal cell line WPMY-1 were established by gene transfection. CCK-8, transwell assays and growth of tumor xenografts were conducted to characterize the effects of CXCL3 on PC-3 cells' proliferation and migration. Western blotting was conducted to test whether CXCL3 could affect the expression of tumorigenesis-associated genes. RESULTS: The results showed that CXCL3 overexpression in PC-3 cells and the PC-3 cells treated with the supernatants of CXCL3-transfected WPMY-1 cells stimulated the proliferation and migration of PC-3 cells in vitro and in a nude mouse xenograft model. Western blotting revealed higher levels of p-ERK, Akt and Bcl-2 and lower levels of Bax in the tumor xenografts transplanted with CXCL3-transfected PC-3 cells. Moreover, the tumor xenografts derived from the PC-3 cells treated with supernatants of CXCL3-transfected WPMY-1 cells showed higher expression of ERK, Akt and Bcl-2 and lower expression of Bax. CONCLUSIONS: These findings suggest that CXCL3 autocrine/paracrine pathways are involved in the development of prostate cancer by regulating the expression of the target genes that are related to the progression of malignancies.
Subject(s)
Cell Movement , Cell Proliferation , Chemokines, CXC/metabolism , Prostate/cytology , Prostatic Neoplasms/metabolism , RNA, Messenger/metabolism , Animals , Autocrine Communication , Cell Line, Tumor , Chemokines, CXC/genetics , Extracellular Signal-Regulated MAP Kinases/genetics , Extracellular Signal-Regulated MAP Kinases/metabolism , Gene Expression , Humans , Male , Mice , Mice, Nude , Neoplasm Transplantation , Paracrine Communication , Phosphorylation , Prostatic Neoplasms/genetics , Prostatic Neoplasms/pathology , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Stromal Cells , Transfection , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolismABSTRACT
Late sodium current (late INa) is enhanced during ischemia by reactive oxygen species (ROS) modifying the Nav 1.5 channel, resulting in incomplete inactivation. Compound 4 (GS-6615, eleclazine) a novel, potent, and selective inhibitor of late INa, is currently in clinical development for treatment of long QT-3 syndrome (LQT-3), hypertrophic cardiomyopathy (HCM), and ventricular tachycardia-ventricular fibrillation (VT-VF). We will describe structure-activity relationship (SAR) leading to the discovery of 4 that is vastly improved from the first generation late INa inhibitor 1 (ranolazine). Compound 4 was 42 times more potent than 1 in reducing ischemic burden in vivo (S-T segment elevation, 15 min left anteriorior descending, LAD, occlusion in rabbits) with EC50 values of 190 and 8000 nM, respectively. Compound 4 represents a new class of potent late INa inhibitors that will be useful in delineating the role of inhibitors of this current in the treatment of patients.
ABSTRACT
Previously we disclosed the discovery of potent Late INa current inhibitor 2 (GS-458967, IC50 of 333nM) that has a good separation of late versus peak Nav1.5 current, but did not have a favorable CNS safety window due to high brain penetration (3-fold higher partitioning into brain vs plasma) coupled with potent inhibition of brain sodium channel isoforms (Nav1.1, 1.2, 1.3). We increased the polar surface area from 50 to 84Å(2) by adding a carbonyl to the core and an oxadiazole ring resulting in 3 GS-462808 that had lower brain penetration and serendipitously lower activity at the brain isoforms. Compound 3 has an improved CNS window (>20 rat and dog) relative to 2, and improved anti-ischemic potency relative to ranolazine. The development of 3 was not pursued due to liver lesions in 7day rat toxicology studies.
Subject(s)
Azoles/pharmacology , Drug Discovery , Heart/drug effects , NAV1.5 Voltage-Gated Sodium Channel/metabolism , Pyridines/pharmacology , Ranolazine/pharmacology , Sodium Channel Blockers/pharmacology , Animals , Azoles/chemical synthesis , Azoles/chemistry , Dogs , Dose-Response Relationship, Drug , Ether-A-Go-Go Potassium Channels/antagonists & inhibitors , Ether-A-Go-Go Potassium Channels/metabolism , Haplorhini , Humans , Molecular Structure , Pyridines/chemical synthesis , Pyridines/chemistry , Rabbits , Ranolazine/chemical synthesis , Ranolazine/chemistry , Rats , Sodium Channel Blockers/chemical synthesis , Sodium Channel Blockers/chemistry , Structure-Activity RelationshipABSTRACT
We started with a medium throughput screen of heterocyclic compounds without basic amine groups to avoid hERG and ß-blocker activity and identified [1,2,4]triazolo[4,3-a]pyridine as an early lead. Optimization of substituents for Late INa current inhibition and lack of Peak INa inhibition led to the discovery of 4h (GS-458967) with improved anti-arrhythmic activity relative to ranolazine. Unfortunately, 4h demonstrated use dependent block across the sodium isoforms including the central and peripheral nervous system isoforms that is consistent with its low therapeutic index (approximately 5-fold in rat, 3-fold in dog). Compound 4h represents our initial foray into a 2nd generation Late INa inhibitor program and is an important proof-of-concept compound. We will provide additional reports on addressing the CNS challenge in a follow-up communication.
Subject(s)
Drug Discovery , Heart/drug effects , NAV1.5 Voltage-Gated Sodium Channel/metabolism , Pyridines/pharmacology , Ranolazine/pharmacology , Sodium Channel Blockers/pharmacology , Triazoles/pharmacology , Animals , Caco-2 Cells , Dose-Response Relationship, Drug , Ether-A-Go-Go Potassium Channels/antagonists & inhibitors , Ether-A-Go-Go Potassium Channels/metabolism , Humans , Macaca fascicularis , Microsomes, Liver/chemistry , Microsomes, Liver/metabolism , Molecular Structure , Pyridines/chemical synthesis , Pyridines/chemistry , Rabbits , Ranolazine/chemical synthesis , Ranolazine/chemistry , Rats , Sodium Channel Blockers/chemical synthesis , Sodium Channel Blockers/chemistry , Structure-Activity Relationship , Triazoles/chemical synthesis , Triazoles/chemistryABSTRACT
PURPOSE: CXCL3 and its receptor CXCR2 were considered to play particularly important roles in the progression of malignancies. However, the investigations about CXCL3/CXCR2 axis in prostate cancer have been poorly involved. Herein we firstly reported our studies on the expression and biological roles of CXCL3 and CXCR2 in prostate cancer. METHODS: Expression levels of CXCL3 and CXCR2 in prostate cancer cell lines (PC-3, DU145 and LNCaP), immortalized prostate stromal cell line (WPMY-1) and immortalized prostate epithelial cell line (RWPE-1) were investigated by RT-PCR, ELISA and western blot, whereas expression levels of CXCL3 in a prostate tissue microarray were detected by immunohistochemistry. Cell counting kit-8 and transwell assays were, respectively, utilized to determine the effects of exogenous CXCL3 on the cell proliferation and migration. We further examined whether CXCL3 could regulate the expression of genes correlated with prostate tumorigenesis by RT- PCR. RESULTS: Elevated expression of CXCR2 was detected in DU145, LNCaP and RWPE-1. Moreover, high-level CXCL3 can be secreted by PC-3 and RWPE-1, and CXCL3 protein expression level in tissue microarray is concordant with prostate cancer metastasis. Exogenous CXCL3 does not contribute to proliferation, but has a significant effect on migration of prostate cancer cells and RWPE-1. Finally, our data showed that exogenous CXCL3 can regulate the expression of genes including ERK, TP73, NUMB, BAX and NDRG3. CONCLUSION: Our findings suggest that CXCL3 and its receptor CXCR2 are overexpressed in prostate cancer cells, prostate epithelial cells and prostate cancer tissues, which may play multiple roles in prostate cancer progression and metastasis.
Subject(s)
Chemokines, CXC/genetics , Gene Expression Regulation, Neoplastic/drug effects , Prostatic Neoplasms/genetics , RNA, Messenger/metabolism , Receptors, Interleukin-8B/genetics , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Chemokines, CXC/metabolism , Chemokines, CXC/pharmacology , Epithelial Cells/metabolism , Humans , Intracellular Signaling Peptides and Proteins , Male , Membrane Proteins/genetics , Nerve Tissue Proteins/genetics , Prostate/cytology , Prostate/metabolism , Prostatic Neoplasms/metabolism , Receptors, Interleukin-8B/metabolism , Stromal Cells/metabolism , Tumor Protein p73/genetics , bcl-2-Associated X Protein/geneticsABSTRACT
Inhibition of cardiac late Na(+) current (I(Na,L)) decreases sodium-dependent calcium overload in diseased hearts. Because INa,L is small in the absence of disease, its inhibition is not expected to significantly alter function of the normal heart. To test this hypothesis, we determined the effects of GS-458967 (GS967), a novel selective inhibitor of I(Na,L) (IC(50) = 0.13 µM), on cardiac function and hemodynamics. The bradycardic agent ivabradine and the Na(+) channel blocker flecainide were used for comparison. A single per os administration of GS967 (5 mg/kg) had no effect on blood pressure or heart rate (HR) in unanesthetized rats. In anesthetized rats, GS967 (0.6 ± 0.1 µM plasma concentration) had no significant effect on HR, PR or QRS electrocardiogram intervals, or contraction. Flecainide (8 mg/kg) slowed HR by 23% ± 3% (P < 0.001), prolonged the PR and QRS intervals by 42% ± 8% and 64% ± 12% (P < 0.001), and had a significant negative inotropic effect. Ivabradine (3 mg/kg) slowed HR by 36% ± 6% (P < 0.001). In rat and rabbit isolated perfused hearts, GS967 (0.1-3 µM) had no significant effects on HR, QRS interval, or contractile function. The results show that selective inhibition of cardiac I(Na,L) is not associated with chronotropic, dromotropic, inotropic, or hemodynamic changes.
Subject(s)
Heart/drug effects , Heart/physiology , Myocardial Contraction/drug effects , Myocardial Contraction/physiology , Sodium Channel Blockers/pharmacology , Animals , Electrophysiological Phenomena/drug effects , Electrophysiological Phenomena/physiology , Female , Heart Rate/drug effects , Heart Rate/physiology , Male , Organ Culture Techniques , Pyridines/pharmacology , Rabbits , Rats , Rats, Sprague-Dawley , Triazoles/pharmacologyABSTRACT
Inhibition of cardiac late sodium current (late I(Na)) is a strategy to suppress arrhythmias and sodium-dependent calcium overload associated with myocardial ischemia and heart failure. Current inhibitors of late I(Na) are unselective and can be proarrhythmic. This study introduces GS967 (6-[4-(trifluoromethoxy)phenyl]-3-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridine), a potent and selective inhibitor of late I(Na), and demonstrates its effectiveness to suppress ventricular arrhythmias. The effects of GS967 on rabbit ventricular myocyte ion channel currents and action potentials were determined. Anti-arrhythmic actions of GS967 were characterized in ex vivo and in vivo rabbit models of reduced repolarization reserve and ischemia. GS967 inhibited Anemonia sulcata toxin II (ATX-II)-induced late I(Na) in ventricular myocytes and isolated hearts with IC(50) values of 0.13 and 0.21 µM, respectively. Reduction of peak I(Na) by GS967 was minimal at a holding potential of -120 mV but increased at -80 mV. GS967 did not prolong action potential duration or the QRS interval. GS967 prevented and reversed proarrhythmic effects (afterdepolarizations and torsades de pointes) of the late I(Na) enhancer ATX-II and the I(Kr) inhibitor E-4031 in isolated ventricular myocytes and hearts. GS967 significantly attenuated the proarrhythmic effects of methoxamine+clofilium and suppressed ischemia-induced arrhythmias. GS967 was more potent and effective to reduce late I(Na) and arrhythmias than either flecainide or ranolazine. Results of all studies and assays of binding and activity of GS967 at numerous receptors, transporters, and enzymes indicated that GS967 selectively inhibited late I(Na). In summary, GS967 selectively suppressed late I(Na) and prevented and/or reduced the incidence of experimentally induced arrhythmias in rabbit myocytes and hearts.
Subject(s)
Anti-Arrhythmia Agents/pharmacology , Arrhythmias, Cardiac/drug therapy , Cardiotonic Agents/pharmacology , Pyridines/pharmacology , Sodium Channel Blockers/pharmacology , Triazoles/pharmacology , Acetanilides/pharmacology , Action Potentials/drug effects , Animals , Arrhythmias, Cardiac/etiology , Arrhythmias, Cardiac/physiopathology , Cnidarian Venoms/pharmacology , Female , Flecainide/pharmacology , Heart Conduction System/drug effects , Long QT Syndrome/genetics , Long QT Syndrome/physiopathology , Mutation/physiology , Myocardial Ischemia/complications , Myocardial Ischemia/physiopathology , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/metabolism , Patch-Clamp Techniques , Piperazines/pharmacology , Potassium Channel Blockers/pharmacology , Quaternary Ammonium Compounds/pharmacology , Rabbits , RanolazineABSTRACT
OBJECTIVE: To explore the influences of milk or coenzyme Q(10) pretreatment to acrylonitrile on vascular endothelial functions in rats. METHODS: A total of 80 rats were randomly divided into 4 groups: control group (Con), acrylonitrile exposure group (ACN), milk pretreatment group (M + ACN) and coenzyme Q(10) pretreatment group (Q(10) + ACN). The experiment was conducted by the method of gavage exposure in rats. Control group was exposed to corn oil; acrylonitrile was administered to other three groups at the doses of 25 mg/kg. The M + ACN and Q(10) + ACN groups were pretreated by milk or coenzyme Q(10) at 30 minutes before acrylonitrile exposure. After a 12-week exposure, the activities of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) were measured in serum and aortal tissues. RESULTS: As compared with Con group [(21.9 ± 1.6) U/ml], the activity of blood serum iNOS was higher in ACN, M + ACN and Q(10) + ACN groups [(42.9 ± 2.5) U/ml, (26.5 ± 4.4) U/ml, (26.7 ± 3.3) U/ml, P < 0.05]. As compared with Con group [(0.540 ± 0.028) U/mg protein], the activity of aortal iNOS was higher in ACN, M + ACN and Q(10) + ACN groups [(0.812 ± 0.008), (0.773 ± 0.019), (0.622 ± 0.013) U/mg protein, (P < 0.05)]. Furthermore the activity of aortal eNOS in Q(10) + ACN group [(0.471 ± 0.011) U/mg protein] was higher than Con, ACN or M + ACN group [(0.371 ± 0.029), (0.380 ± 0.016), (0.425 ± 0.020) U/mg protein, P < 0.05]. CONCLUSION: Chronic administration of ACN by gavages results in vascular endothelial dysfunctions. Milk and coenzyme Q(10) pretreatment reduce this effect in rats.
Subject(s)
Acrylonitrile/toxicity , Endothelium, Vascular/physiopathology , Milk , Ubiquinone/analogs & derivatives , Animals , Endothelium, Vascular/metabolism , Female , Male , Nitric Oxide Synthase Type II/metabolism , Nitric Oxide Synthase Type III/metabolism , Rats , Rats, Sprague-Dawley , Toxicity Tests , Ubiquinone/pharmacologyABSTRACT
OBJECTIVE: To explore the postoperative visual acuity results of wavefront-guided LASIK with iris recognition for myopia or myopic astigmatism and the changes of higher-order aberrations and contrast sensitivity function (CSF). METHODS: Series of prospective case studies, 158 eyes (85 cases) of myopia or myopic astigmatism were divided into two groups: one group underwent wavefront-guided LASIK with iris recognition (iris recognition group); another group underwent wavefront-guided LASIK treatment without iris recognition through the limbus maring point (non-iris recognition group). To comparative analyze the postoperative visual acuity, residual refraction, the RMS of higher-order aberrations and CSF of two groups. RESULTS: There was no statistical significance difference between two groups of the average uncorrected visual acuity (t = 0.039, 0.058, 0.898; P = 0.844, 0.810, 0.343), best corrected visual acuity (t = 0.320, 0.440, 1.515; P = 0.572, 0.507, 0.218), and residual refraction [spherical equivalent (t = 0.027, 0.215, 0.238; P = 0.869, 0.643, 0.626), spherical (t = 0.145, 0.117, 0.038; P = 0.704, 0.732, 0.845) and cylinder (t = 1.676, 1.936, 0.334; P = 0.195, 0.164, 0.563)] at postoperative 10 days, 1 month and 3 month. The security index of iris recognition group at postoperative 3 month was 1.06 and non-iris recognition group was 1.03; the efficacy index of iris recognition group is 1.01 and non-iris recognition group was 1.00. Postoperative 3 month iris recognition group 93.83% eyes and non-iris recognition group of 90.91% eyes spherical equivalent within ± 0.50 D (χ(2) = 0.479, P = 0.489), iris recognition group of 98.77% eyes and non-iris recognition group of 97.40% eyes spherical equivalent within ± 1.00 D (Fisher test, P = 0.613). There was no significance difference between the two groups of security, efficacy and predictability. Non-iris recognition group postoperative 1 month and postoperative 3 months 3-order order aberrations root mean square value (RMS) higher than the iris recognition group increased (t = 3.414, -2.870; P = 0.027, 0.045), in particular of coma; the general higher-order aberrations (t = 0.386, 1.132; P = 0.719, 0.321), 4-order aberrations (t = 0.808, 2.720; P = 0.464, 0.063), and 5-order aberrations (t = 0.148, -1.717; P = 0.890, 0.161) show no statistically significant difference. Three months after surgery, two groups have recovered at all spatial frequencies of CSF, iris recognition group at 3.0 c/d (t = 3.209, P = 0.002) and 6.0 c/d (t = 2.997, P = 0.004) spatial frequencies of CSF under mesopic condition was better than non-iris recognition group, glare contrast sensitivity function (GCSF) for 3.0 c/d (t = 3.423, P = 0.001) and 6.0 c/d (t = 6.986, P = 0.000) spatial frequencies under mesopic condition and 1.5 c/d (t = 9.839, P = 0.000) and 3.0 c/d (t = 7.367, P = 0.000) spatial frequencies under photopic condition in iris recognition group were better than non-iris recognition group, there were no significant difference between two groups at the other spatial frequencies. CONCLUSIONS: Wavefront-guided LASIK with or without iris recognition both acquired better postoperative visual acuity, but in comparison with without iris recognition, wavefront-guided LASIK with iris recognition is efficient to reduce coma and enhance contrast sensitivity of postoperative.
Subject(s)
Astigmatism/surgery , Iris , Keratomileusis, Laser In Situ/methods , Myopia/surgery , Adolescent , Adult , Contrast Sensitivity , Female , Humans , Male , Myopia/complications , Prospective Studies , Treatment Outcome , Visual Acuity , Young AdultABSTRACT
We tested the effect of the antianginal agent ranolazine on ventricular arrhythmias in an ischemic model using two protocols. In protocol 1, anesthetized rats received either vehicle or ranolazine (10 mg/kg, iv bolus) and were subjected to 5 min of left coronary artery (LCA) occlusion and 5 min of reperfusion with electrocardiogram and blood pressure monitoring. In protocol 2, rats received either vehicle or three doses of ranolazine (iv bolus followed by infusion) and 20 min of LCA occlusion. With protocol 1, ventricular tachycardia (VT) occurred in 9/12 (75%) vehicle-treated rats and 1/11 (9%) ranolazine-treated rats during reperfusion (P = 0.003). Sustained VT occurred in 5/12 (42%) vehicle-treated but 0/11 in ranolazine-treated rats (P = 0.037). The median number of episodes of VT during reperfusion in vehicle and ranolazine groups was 5.5 and 0, respectively (P = 0.0006); median duration of VT was 22.2 and 0 s in vehicle and ranolazine rats, respectively (P = 0.0006). With protocol 2, mortality in the vehicle group was 42 vs. 17% (P = 0.371), 10% (P = 0.162) and 0% (P = 0.0373) with ranolazine at plasma concentrations of 2, 4, and 8 microM, respectively. Ranolazine significantly reduced the incidence of ventricular fibrillation [67% in controls vs. 42% (P = 0.414), 30% (P = 0.198) and 8% (P = 0.0094) in ranolazine at 2, 4, and 8 microM, respectively]. Median number (2.5 vs. 0; P = 0.0431) of sustained VT episodes, incidence of sustained VT (83 vs. 33%, P = 0.0361), and the duration of VT per animal (159 vs. 19 s; P = 0.0410) were also significantly reduced by ranolazine at 8 microM. Ranolazine markedly reduced ischemia-reperfusion induced ventricular arrhythmias. Ranolazine demonstrated promising anti-arrhythmic properties that warrant further investigation.
Subject(s)
Acetanilides/pharmacology , Anti-Arrhythmia Agents/pharmacology , Myocardial Ischemia/drug therapy , Myocardial Reperfusion Injury/drug therapy , Piperazines/pharmacology , Tachycardia, Ventricular/prevention & control , Ventricular Fibrillation/prevention & control , Acetanilides/administration & dosage , Acetanilides/blood , Angina Pectoris/drug therapy , Animals , Anti-Arrhythmia Agents/administration & dosage , Anti-Arrhythmia Agents/blood , Blood Pressure/drug effects , Disease Models, Animal , Dose-Response Relationship, Drug , Electrocardiography , Female , Heart Rate/drug effects , Infusions, Intravenous , Injections, Intravenous , Male , Myocardial Ischemia/complications , Myocardial Ischemia/physiopathology , Myocardial Reperfusion Injury/complications , Myocardial Reperfusion Injury/physiopathology , Piperazines/administration & dosage , Piperazines/blood , Ranolazine , Rats , Rats, Sprague-Dawley , Tachycardia, Ventricular/etiology , Tachycardia, Ventricular/physiopathology , Time Factors , Ventricular Fibrillation/etiology , Ventricular Fibrillation/physiopathologyABSTRACT
OBJECTIVE: To clone and express surface antigen SAG4 gene of Toxoplasma gondii, and analyze its immunoreactivity. METHODS: Specific primers were designed based on the reported SAG4 gene of T. gondii RH strain (GenBank Accession No: AF340224.1). Using genomic DNA from T. gondii as templates, SAG4 gene was amplified by PCR. The PCR product was cloned into pMD19-T vector and identified by digestion with restriction enzyme and PCR. Then the target fragment was subcloned into pET28a(+) vector, transformed into E. coli BL21 and followed by expression of the protein induced by IPTG. The protein was identified by Western blotting. RESULTS: The target gene was amplified with the length of 537 bp. Sequence analysis showed that the predicted amino acid sequence was identical with that of SAG4 as a membrane protein in T. gondii. After induced by IPTG, the recombinant SAG4 protein existed in an inclusion body form. The recombinant SAG4 (Mr 18 740) was recognized by serum of infected mice. CONCLUSION: SAG4 has been expressed and shows certain immuno-response activity.
Subject(s)
Antigens, Protozoan/genetics , Membrane Glycoproteins/immunology , Protozoan Proteins/immunology , Toxoplasma/genetics , Animals , Antigens, Protozoan/immunology , Antigens, Surface/genetics , Antigens, Surface/immunology , Blotting, Western , Cloning, Molecular , Female , Membrane Glycoproteins/genetics , Mice , Protozoan Proteins/genetics , Toxoplasma/immunologyABSTRACT
BACKGROUND: Impaired brachial flow-mediated dilation (FMD) and increased carotid intima-media thickness (IMT) are associated with increased risk of cardiovascular events. METHODS: We measured brachial FMD and a mean of 12 sites maximum carotid IMT (mmIMT) in 279 patients (mean age 62 +/- 12 y; 163 men) admitted for coronary angiography due to chest pain. HYPOTHESIS: There are gender differences in the predictive values of FMD and IMT for cardiovascular events. RESULTS: Univariable analysis showed that impaired FMD (p < 0.001), but not increased mmIMT (p = 0.056), significantly predicted spontaneous cardiovascular events. After adjusting for the extent of coronary artery disease (CAD) and other clinical variables, age (heart rate [HR] 1.05, 95% confidence interval [CI]: 1.01-1.09, p = 0.017) and FMD (HR 0.85, 95% CI: 0.75-0.97, p = 0.012) were independent predictors for cardiovascular events. A total of 148 (53%) patients had CAD (> or =50% diameter stenosis). Over a median follow-up of 16 mo, 36 (12.9%) patients experienced spontaneous cardiovascular events (cardiovascular death, stroke, acute myocardial infarction [MI], unstable angina pectoris, and congestive heart failure [HF]). Women were more likely than men to develop cardiovascular events in patients without significant CAD (11.9% versus 1.6%, odds ratio [OR] = 8.54, p = 0.033), but not in those patients with CAD (20.4 % versus 17.2%, OR = 1.24, p = 0.66). Moreover, women accounted for 8 (88.9%) events in non-CAD patients. Furthermore, impaired FMD predicted the occurrence of cardiovascular events in both men and women (p < 0.05). CONCLUSION: Brachial FMD, rather than carotid IMT, was an independent predictor for cardiovascular events after adjusting for the extent of CAD. Moreover, impaired brachial endothelial function in women without significant CAD was associated with an increased risk of spontaneous cardiovascular events.