Alu antisense RNA ameliorates methylglyoxal-induced human lens epithelial cell apoptosis by enhancing antioxidant defense.

AIM: To determine whether an antisense RNA corresponding to the human Alu transposable element (Aluas RNA) can protect human lens epithelial cells (HLECs) from methylglyoxal-induced apoptosis. METHODS: Cell counting kit-8 (CCK-8) and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were used to assess HLEC viability. HLEC viability/death was detected using a Calcein-AM/PI double staining kit; the annexin V-FITC method was used to detect HLEC apoptosis. The cytosolic reactive oxygen species (ROS) levels in HLECs were determined using a reactive species assay kit. The levels of malondialdehyde (MDA) and the antioxidant activities of total-superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) were assessed in HLECs using their respective kits. RT-qPCR and Western blotting were used to measure mRNA and protein expression levels of the genes. RESULTS: Aluas RNA rescued methylglyoxal-induced apoptosis in HLECs and ameliorated both the methylglyoxal-induced decrease in Bcl-2 mRNA and the methylglyoxal-induced increase in Bax mRNA. In addition, Aluas RNA inhibited the methylglyoxal-induced increase in Alu sense RNA expression. Aluas RNA inhibited the production of ROS induced by methylglyoxal, restored T-SOD and GSH-Px activity, and moderated the increase in MDA content after treatment with methylglyoxal. Aluas RNA significantly restored the methylglyoxal-induced down-regulation of Nrf2 gene and antioxidant defense genes, including glutathione peroxidase, heme oxygenase 1, γ-glutamylcysteine synthetase and quinone oxidoreductase 1. Aluas RNA ameliorated methylglyoxal-induced increases of the mRNA and protein expression of Keap1 that is the negative regulator of Nrf2. CONCLUSION: Aluas RNA reduces apoptosis induced by methylglyoxal by enhancing antioxidant defense.

Reactive arthritis and subacute infective endocarditis caused by Streptococcus gordonii infection: A case report.

Streptococcus gordonii (S. gordonii) belongs to the alpha-hemolytic Streptococcus group. It is a symbiotic bacterium found in the human oral mucosa which is present in large quantities on the surface of the teeth. It is generally considered nonpathogenic or weakly pathogenic and is known to cause subacute endocarditis; however, there are few reports of reactive arthritis (ReA) caused by S. gordonii. Herein, we report a case of ReA complicated by subacute infective endocarditis caused by S. gordonii and explore the possible pathogenic mechanism of ReA caused by S. gordonii.

The Dynamics of Circulating Monocyte Subsets and Intra-Plaque Proliferating Macrophages during the Development of Atherosclerosis in ApoE-/- Mice.

To detect the development of monocytes and proliferative macrophages in atherosclerosis of ApoE-/- mice, we randomly assigned 84 ApoE-/- mice fed western diet or chow diet. On weeks 2, 4, 6, 8, 10, and 12 after fed high-fat diet or normal chow diet, animals were euthanized (n = 7 for each group at each time point). Flow cytometry methods were used to analyze the proportions of circulation monocyte subsets. The macrophage and proliferative macrophage accumulation within atherosclerotic plaques was estimated by confocal florescence microscopy. Plasma levels of total cholesterol and triglyceride were measured by ELISA kit. The plaques of aortic sinus were stained with Oil Red O. The percent of Ly6Chi circulation monocyte, the density of proliferation macrophage, the total plasma cholesterol and triglyceride levels, the lesion area of ApoE-/- mice were consistently elevated in chow diet throughout the trial. The total plasma cholesterol and triglyceride levels, the lesion area were elevated in western diet group with age, and they were always higher than the chow diet group. The Ly6Chi monocytes and proliferative macrophages reached a plateau at 8 weeks and 6 weeks; despite continued high-triglyceride high-cholesterol diet the percent did not significantly change. Interestingly, the density of macrophage did not change significantly over age in western and chow diet groups. Our results provide a dynamic view of Ly6Chi monocyte subset, the density of macrophage and proliferation macrophage change during the development and progression of atherosclerosis, which is relevant for designing new treatment strategies targeting mononuclear phagocytes in this model.

Trajectories of Circulating Monocyte Subsets After ST-Elevation Myocardial Infarction During Hospitalization: Latent Class Growth Modeling for High-Risk Patient Identification.

It remains unclear if the developmental trajectories of a specific inflammatory biomarker during the acute phase of ST-elevation myocardial infarction (STEMI) provide outcome prediction. By applying latent class growth modeling (LCGM), we identified three distinctive trajectories of CD14++CD16+ monocytes using serial flow cytometry assays from day 1 to day 7 of symptom onset in 96 de novo STEMI patients underwent primary percutaneous coronary intervention. Membership in the high-hump-shaped trajectory (16.8%) independently predicted adverse cardiovascular outcomes during a median follow-up of 2.5 years. Moreover, inclusion of CD14++CD16+ monocyte trajectories significantly improved area under the curve (AUC) when added to left ventricular ejection fraction-based prediction model (ΔAUC = 0.093, P = 0.013). Therefore, CD14++CD16+ monocyte trajectories during STEMI hospitalization are a novel risk factor for post-STEMI adverse outcomes. These results provide the first proof-of-principle evidence in support of the risk stratification role of LCGM-based longitudinal modeling of specific inflammatory markers during acute STEMI.

Isolation and identification of anti-tumor polysaccharide LSP21 from Limonium sinense (Girard) Kuntze.

Limonium sinense (Girard) Kuntze is a traditional Chinese folk medicine used for the treatment of fever, hemorrhage, hepatitis and other disorders. Recently, it was found that the crude polysaccharides from L. sinense (LSP) has significant anti-tumor activity. However, research on the isolation and identification of anti-tumor polysaccharide fractions from LSP has not yet been reported. In this study, three polysaccharides LSP11, LSP21, LSP31 were isolated and purified from LSP by using DEAE-52 cellulose column and Sephadex G-100 column chromatography. It was found that LSP21 exhibited the most significant inhibitory effect on the growth of HepG2 cells in vitro. Further research showed that LSP21 inhibited the growth of HepG2 cells in a dose-dependent manner and could induce cell body shrinkage, chromatin condensation, and reduction in the number of tumor cells with normal morphology which suggested that its cytotoxicity on tumor cell might be related to both inhibition on cell proliferation and inducement of cell death. Finally, the structural characteristics of LSP21 were analyzed by high performance liquid chromatography (HPLC) and gas chromatography (GC). The results showed that LSP21 is a heteropolysaccharide with an average molecular weight of 1.31×10(6) Da and consists of glucose, galactose and mannose in the ratio of 1.77:1:2.38.

Antitumor and immunomodulatory activity of polysaccharides from the root of Limonium sinense Kuntze.

Limonium sinense (Girard) Kuntze is a traditional Chinese folk medicine used for the treatment of fever, hemorrhage, hepatitis and other disorders. The study focused on the antitumor and immunomodulatory activities of L. sinense polysaccharides (LSP) which was obtained from the root of the plant. The antitumor effects of only LSP and LSP in combination with 5-fluorouracil (5-FU) were both evaluated with Heps-bearing tumor mice models. In addition, the macrophage phagocytosis assay, splenocyte proliferation and cytokines production tests were used to assess the immunomodulatory activities of LSP. The results revealed that the LSP (at the dose of 200 and 400 mg/kg) had an obvious inhibition on the growth of transplanted mouse tumor. It also exhibited a significant synergistic effect of antitumor activity when combined with 5-FU (p<0.05). Furthermore, the LSP (at the dose of 100 and 200 mg/kg) remarkably improved macrophage phagocytosis function in immune suppressed mice. In addition, LSP (at the dose of 50-200 µg/ml) showed significant synergistic effects on ConA-stimulated proliferation and IFN-γ and IL-2 production of splenocyte in vitro (p<0.05). These findings suggest that LSP had clear antitumor activity which might be related to its regulation of immune function in mice.

Preservation on calcium homeostasis is involved in mitochondrial protection of Limonium sinense against liver damage in mice.

Mechanisms underlying the mitochondrial protection of Limonium sinense extracts (LSE) was studied in lipopolysaccharide and D-galactosamine (LPS/D-GalN) intoxicated mice. It was found that increased activities of serum aspartate aminotransferase and alanine aminotransferase induced by LPS/D-GalN were significantly inhibited by pretreatment with LSE. The obvious disruption of membrane potential, intramitochondrial Ca (2+) overload and suppression in mitochondrial Ca (2+) -ATPase activity induced by LPS/D-GalN were significantly blocked by pretreatment with LSE. It was concluded that mechanisms underlying protection of LSE against liver mitochondria damage might be related to the preservation on mitochondrial Ca (2+) homeostasis through the preservation on mitochondrial Ca (2+) -ATPase activity.