Common MIR146A Polymorphisms in Chinese Ankylosing Spondylitis Subjects and Controls

<div><p>Common polymorphisms of microRNA gene MIR146A were reported as associated with different autoimmune diseases, include systemic lupus erythematosus, psoriatic arthritis, asthma and ankylosing spondylitis. In this study we investigated MIR146A SNPs in Chinese people with ankylosing spondylitis. Three common SNPs: rs2910164, rs2431697 and rs57095329 were selected and genotyped in 611 patients and 617 controls. We found no association between these SNPs and ankylosing spondylitis in our samples.</p></div

MIR146A gene SNPs distribution in AS and control subjects.

<p>MIR146A gene SNPs distribution in AS and control subjects.</p

Expansion of Tfh cells in MRL/lpr mice.

<p>(<b>A</b>) Splenomegaly in MRL/lpr mice. (<b>B</b>) Splenocytes were isolated from MRL/lpr and B6 mice. After staining, cells were first gated for CD4⁺ T cells, and the CXCR5⁺PD-1⁺ cells were analyzed with a CD4⁺ gate by flow cytometry. (<b>C</b>) The percentages of CXCR5⁺PD-1⁺ cells among CD4⁺ T cells (n = 6 for each group). (<b>D</b>) IL-21 mRNA expression in fresh isolated splenocytes was determined by real-time RT-PCR (n = 6 for each group). (<b>E</b>) Bcl-6 mRNA expression in fresh isolated splenocytes was determined by real-time RT-PCR (n = 6 for each group). (<b>F</b>) Sorted CD4⁺CXCR5⁺PD-1⁺ Tfh cells from MRL/lpr and B6 mice were cultured in the presence of anti-CD3 and anti-CD28 for 2 days, IL-21 mRNA expression was determined by real-time RT-PCR. Results shown are representative of at least three independent experiments. (<b>G</b>) Sorted CD4⁺CXCR5⁺PD-1⁺ Tfh cells from MRL/lpr and B6 mice were cultured in the presence of anti-CD3 and anti-CD28 for 2 days, Bcl-6 mRNA expression was determined by real-time RT-PCR. Results shown are representative of at least three independent experiments. (<b>H</b>) IL-21 expression in spleens was confirmed by immunohistochemical staining. Further magnification of the black-bordered box shows the predominance of IL-21⁺ lymphocytes. The scale bar represents 50 μm.</p

IL-21 induces IL-10 production during the differentiation of B10 cells via activation of p-STAT3.

<p>(<b>A</b>) Naïve B cells sorted from B6 mice were cultured in the presence of LPS with or without 10 ng/ml IL-21 for the indicated times and stimulated with PIB for the final 5 hours. STAT3 mRNA expression was detected by real-time RT-PCR. (<b>B</b>) The expression of p-STAT3 and STAT3 in sorted CD19⁺CD5⁺CD1d^high B cells from MRL/lpr mice and B6 mice was analyzed by Western blot. (<b>C</b>) Naïve B cells sorted from B6 mice were cultured with or without 10 ng/ml IL-21 for the indicated times. Then, p-STAT3 proteins were analyzed by Western blot. (<b>D</b>) Naïve B cells sorted from B6 mice were cultured with or without 10 ng/ml IL-21 or AG490 for the indicated times, and the levels of p-STAT3 and STAT3 proteins were determined by Western blot. (<b>E</b>) Naïve B cells sorted from B6 mice were cultured in the presence of LPS with or without 10 ng/ml IL-21 or AG490 for the indicated times and stimulated with PI for the final 5 hours. IL-10 in supernatants was detected by ELISA. (<b>F</b>) Naïve B cells sorted from B6 mice were cultured in the presence of LPS with or without the indicated concentrations of IL-21 or AG490 for 48 hours and stimulated with PI for the final 5 hours. IL-10 in supernatants was detected by ELISA. (<b>G</b>) Sorted CD19⁺CD5⁺CD1d^high B cells from MRL/lpr mice were cultured in the presence of LPS and 10 ng/ml IL-21 with or without AG490 for 48 hours, and IL-10 in supernatants was detected by ELISA. (<b>H</b>) Naïve B cells sorted from B6 mice were cultured with or without 10 ng/ml IL-21 or SPI for 15 minutes, p-STAT3 proteins were then analyzed by Western blot. (<b>I</b>) Naïve B cells sorted from B6 mice were cultured in the presence of LPS with or without 10 ng/ml IL-21 or SPI for the indicated times and stimulated with PI for the final 5 hours, IL-10 in supernatants was then detected by ELISA. (<b>J</b>) CFSE-labeled CD4⁺CD25^- T cells from MRL/lpr mice were cultured for 3 days with anti-CD3 and anti-CD28 antibodies in combination with 20% of the supernatants from IL-21-induced B10 cell cultures (from MRL/lpr mice) with or without neutralization of IL-10. The proliferation of these cells was determined by flow cytometry. All the above results shown are representative of at least three independent experiments.</p

Tfh cells are associated with autoantibody production in MRL/lpr mice.

<p>(<b>A</b>) H&E staining of spleens from MRL/lpr and B6 mice (left). PNA⁺ GC cells were determined by immunohistochemical staining (right). The scale bar represents 50 μm. (<b>B</b>) A positive correlation between the percentages of CD4⁺CXCR5⁺PD-1⁺ T cells and the number of PNA⁺ GC cells in spleens of MRL/lpr mice (n = 6) was observed. (<b>C</b>) A positive correlation between the percentages of CD4⁺CXCR5⁺PD-1⁺ T cells and renal scores of MRL/lpr mice (n = 6) was observed. (<b>D</b>) A positive correlation between serum levels of IL-21 and ANA in MRL/lpr mice (n = 6) was found. (<b>E</b>) A positive correlation between serum levels of IL-21 and ds-DNA in MRL/lpr mice (n = 6) was found. (<b>F</b>) Sorted CD4⁺CXCR5⁺PD-1⁺ Tfh cells from MRL/lpr and B6 mice were cultured in the presence of anti-CD3 and anti-CD28 for 2 days, detection of IL-21 in the supernatants by ELISA. Results shown are representative of at least three independent experiments. (<b>G</b>) The concentrations of IgM and IgG₁ in sorted naive B cells from B6 mice following a 3-day induction with LPS, anti-CD40, anti-IgM, and 20% of the supernatants from cultured Tfh cells from MRL/lpr (mTfh) and B6 mice (bTfh) or vehicle (culture media with 2 µg/ml plate-bound anti-CD3 and 2 µg/ml soluble anti-CD28) with or without neutralization of IL-21. Results shown are representative of at least three independent experiments.</p

Table1_Comparison of the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells and adipose-derived stem cells on erectile dysfunction in a rat model of bilateral cavernous nerve injury.DOCX

Background: Cavernous nerve injury (CNI) is the leading cause of erectile dysfunction (ED) after radical prostatectomy and pelvic fracture. Transplantation of human adipose-derived stem cells (ASCs) has been widely used to restore erectile function in CNI-ED rats and patients. Umbilical cord blood-derived MSCs (CBMSCs) are similarly low immunogenic but much primitive compared to ASCs and more promising in large-scale commercial applications due to the extensive establishment of cord blood banks. However, whether CBMSCs and ASCs have differential therapeutic efficacy on CNI-ED and the underlying mechanisms are still not clear.Materials and methods: A bilateral cavernous nerve injury (BCNI) rat model was established by crushing the bilateral cavernous nerves. After crushing, ASCs and CBMSCs were intracavernously injected immediately. Erectile function, Masson staining, and immunofluorescence analyses of penile tissues were assessed at 4 and 12 weeks. PKH-26-labeled ASCs or CBMSCs were intracavernously injected to determine the presence and differentiation of ASCs or CBMSCs in the penis 3 days after injection. In vitro experiments including intracellular ROS detection, mitochondrial membrane potential assay, EdU cell proliferation staining, cell apoptosis assay, and protein chip assay were conducted to explore the underlying mechanism of CBMSC treatment compared with ASC treatment.Results: CBMSC injection significantly restored erectile function, rescued the loss of cavernous corporal smooth muscles, and increased the ratio of smooth muscle to collagen. PKH-26-labeled CBMSCs or ASCs did not colocalize with endothelial cells or smooth muscle cells in the corpus cavernosum. Moreover, the conditioned medium (CM) of CBMSCs could significantly inhibit the oxidative stress and elevate the mitochondria membrane potential and proliferation of Schwann cells. Better therapeutic effects were observed in the CBMSC group than the ASC group both in vivo and in vitro. In addition, the content of neurotrophic factors and matrix metalloproteinases in CBMSC-CM, especially NT4, VEGF, MMP1, and MMP3 was significantly higher than that of ASC-CM.Conclusion: Intracavernous injection of CBMSCs exhibited a better erectile function restoration than that of ASCs in CNI-ED rats owing to richer secretory factors, which can promote nerve regeneration and reduce extracellular matrix deposition. CBMSC transplantation would be a promising therapeutic strategy for CNI-ED regeneration in the future.</p

Additional file 1: of Physiological, hematological and biochemical factors associated with high-altitude headache in young Chinese males following acute exposure at 3700Â m

The distribution and QQ-norm plot of SpO2 at 50 m and 3700 m. (DOCX 168 kb

Additional file 2: of Physiological, hematological and biochemical factors associated with high-altitude headache in young Chinese males following acute exposure at 3700Â m

The incidence of mild, moderate and severe headaches after ascent to 3700 m altitude. (DOCX 16 kb

Additional file 3: of Physiological, hematological and biochemical factors associated with high-altitude headache in young Chinese males following acute exposure at 3700Â m

The Shapiro-Wilk normality test of parameters at 50 m and 3700 m. (DOCX 12 kb

Le Courrier

18 septembre 18371837/09/18 (N261)