Effects of dexamethasone on MRL/Ipr mice with systemic lupus erythematosus complicated with cognitive dysfunction / 中南大学学报(医学版)

Objective:To evaluate the effects of dexamethasone on systemic lupus erythematosus complicated with cognitive dysfunction.Methods:Ten wild type mice and 20 MRL/lpr mice were applied for the research.MRL/lpr mice were randomly assigned to a MRL/lpr group and a MRL/lpr + dexamethasone (1.5 mg/kg) group.Interleukin-6 (IL-6),IL-1β,and tumor necrosis factor alpha (TNF-α) in serum and hippocampus were detected.The protein phosphorylation levels of phosphoinositide 3-kinase (P-PI3K),protein kinase B (P-Akt),NF-kappa-B inhibitor alpha (P-IκBa) and nuclear transcription factor kappa-B p65 (P-NF-κB p65) were detected by Western blot,the level of P-NF-κB p65 also was detected by immunohistochemistry.Results:Treatment with dexamethasone (1.5 mg/kg) alleviated the cognitive dysfunction and decreased the levels of IL-6,IL-1 β and TNF-α in serum and hippocampus,and reduced the levels of P-PI3K,P-Akt,P-IκBa and P-NF-κB p65 in hippocampus in MRL/lpr mice.Conclusion:Dexamethasone may play a protective role in the cognitive function by decreasing the levels of TNF-α and IL-1 β in the hippocampus of MRL/lpr lupus mice.

Peripheral blood T cell TNF-α and IFN-γ production stimulated by low molecular peptide of Mycobacterium tuberculosis heat-resistant antigen for differential diagnosis between pulmonary tuberculosis and latent tuberculosis infection / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of low molecular peptide of Mycobacterium tuberculosis heat-resistant antigen (Mtb-HAg-10k) on the production of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in peripheral blood T cells and test the feasibility of differential diagnosis between pulmonary tuberculosis (PTB) and latent tuberculosis infection (LTBI) by assessing the number of Mtb-HAg-10k-stimulated IFN-γ-producing T cells.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMCs) were separated from the peripheral blood of 10 healthy adults, 6 individuals with LTBI and 13 patients with PTB. The PBMCs were cultured in the presence of Mtb-HAg-10k obtained by ultrafiltration centrifugation, with Mtb-HAg and phytohaemagglutinin (PHA) as the controls. The proportions of TNF-α- and IFN-γ-producing cells in the T cell subsets were detected by flow cytometry (FCM), and the number of IFN-γ-producing cells from patients with PTB and LTBI was detected with ELISPOT.</p><p><b>RESULTS</b>Flow cytometry showed that Mtb-HAg-10k exposure resulted in a significantly higher proportion of TNF-α-producing γδT cells than that of IFN-γ-producing γδT cells in the PBMCs (P<0.01). Compared with the PBMCs exposed to PHA, the PBMCs exposed to Mtb-HAg-10k exhibited a significantly greater proportion of γδT cells that produced both TNF-α and IFN-γ (P<0.01) but a significantly lower proportion of αβT cells producing both TNF-α and IFN-γ (P<0.01). Mtb-HAg-10k exposure of the PBMCs caused a significant reduction in the number of IFN-γ-producing cells as compared with Mtb-HAg and PHA treatments (P<0.01), and this reduction was more obvious in PBMCs from patients with PTB than in those from individuals with LTBI (P<0.01).</p><p><b>CONCLUSION</b>Mtb-HAg-10k can markedly induce γδT cells in the PBMCs to produce TNF-α and IFN-γ, and detection of the number of IFN-γ-producing cells in the PBMCs following Mtb-HAg-10k stimulation helps in the differential diagnosis between pulmonary tuberculosis and latent tuberculosis infection.</p>

Simultaneous determination of 3-chlorotyrosine and 3-nitrotyrosine in human plasma by direct analysis in real time-tandem mass spectrometry

A novel method for the simultaneous determination of 3-nitrotyrosine (NT) and 3-chlorotyrosine (CT) in human plasma has been developed based on direct analysis in real time-tandem mass spectrometry (DART-MS/MS). Analysis was performed in the positive ionization mode using multiple reaction monitoring (MRM) of the ion transitions at m/z 216.2/170.1 for CT, m/z 227.2/181.1 for NT and m/z 230.2/184.2 for the internal standard, d (3)-NT. The assay was linear in the ranges 0.5-100 μg/mL for CT and 4-100 μg/mL for NT with corresponding limits of detection of 0.2 and 2 μg/mL. Intra- and inter-day precisions and accuracies were respectively <15% and ±15%. Matrix effects were also evaluated. The method is potentially useful for high throughput analysis although sensitivity needs to be improved before it can be applied in clinical research.

Optimization of process variables for the manganese peroxidase of the white-rot fungus Schizophyllum sp. F17 by full factorial central composite design / 生物工程学报

White-rot fungus manganese peroxidase (MnP) that has great potential in degrading azo dyes is one of the extracellular glycolsylated heme proteins. MnP from Schizophyllum sp. F17 was isolated and purified by Sephadex G-75 gel filtration chromatography followed by DEAE-cellulose anion exchange chromatography. The molecular weight of the puried enzyme was 49.2 kDa, while the half-life of the MnP in the presence of 0.1 mmol/L H2O2 was 5-6 min. The efficiency of MnP-catalyzed reactions were determined by three key factors: the concentrations of Mn2+, H2O2, and the amount of MnP. Using single factor analysis, an optimized concentration of Mn2+, H2O2 and enzyme were optimized to be 1.2 mmol/L, 0.1 mmol/L, and 0.4 mL, respectively. A response surface methodology (RSM) employing two-level-three-factor full factorial central composite design was used to optimize the catalytic conditions. The result showed that the concentration of H2O2 and the interaction between H2O2 and MnP mostly affect the MnP catalytic efficiency. Finally, we show that the azo dyes could be efficiently decolorized by the purified MnP under optimized conditions.

Effect of Tenuigenin on Mash1 expression and directional differentiation of neural stem cells from mouse hippocampus in vitro / 中华神经医学杂志

Objective To determine the effect of Tenuigenin (TEN) on the Mashl expression in neural stem cells (NSCs) from mouse hippocampus and directional differentiation of its NSCs in vitro. Methods NSCs were generated from the mouse hippocampus within 24 hours after birth and cultured with EGF and bFGF (20 ng/mL, each) in a 50-mm uncoated culture flask. The third-passage NSCs were cultured with TEN medium and divided them into vehicle, 5 mg/L, 20 mg/L and 100 mg/L groups according to different TEN concentrations. Immunocytochemical techniques were employed to detect the NSE-positive neurons and ChAT-positive neurons induced by TEN, and the expression of Mash1 in NSCs affected by TEN was measured by RT-PCR techniques. Results The immunocytochemical and RT-PCR results revealed that ChAT-positive neurons and NSE-positive neurons were detected with statistical significance and 20 mg/L group showed a stronger effect than other concentration groups (P< 0.05). The expression of Mash1 in the 20 mg/L group increased as compared with that in the vehicle group (P<0.05). Conclusion TEN can induce NSCs in vitro differentiating into neurons and cholinergic neurons, and increase the expression of Mashl in NSCs.