Granulocyte-macrophage colony-stimulating factor effectively induces CD14+ HLA-DR-iNOS+ myeloid derived suppressor cells from peripheral blood monocytes / 中华器官移植杂志

Objective To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the generation of human myeloid derived suppressor cells (MDSCs) relied on peripheral blood monocytes,and to establish efficient induction system in vitro of MDSCs.Methods Kidney transplantation recipients between January and March 2017 were included in this study.Purified CD14 + cells isolated from peripheral blood were cultured in the presence of GM-CSF with different concentrations for 7 days.Phenotypes and immunosuppressive abilities of induced MDSCs (iMDSCs) were investigated with FACS analyses.Inducible nitric oxide synthase (iNOS) mRNA expression was detected by qRT-PCR to determine the influence of iNOS-pathway on the immunosuppressive abilities of iMDSCs.Results A total of 11 recipients were included in this study.HLA-DR expression decreased sharply after the culture with GM-CSF.iMDSCs showed the similar phenotype characteristics with monocytic-MDSCs (M-MDSCs) as well as significant ability to suppress T cells proliferation and cytokines production.iMDSCs expressed higher levels of iNOS than monocytes,and the inhibitor effects of iMDSCs were significantly reduced after treatment with L-NMMA (1 mmol/L).The variations of phenotype and suppressive ability were concentrationdependent,and more significant changes could be revealed in the group of 10 μg/L GM-CSF.Conclusion GM-CSF-treated peripheral blood monocytes can be efficiently transformed to M-MDSCs,and suppress T cells proliferation and cytokines secretion via iNOS-dependent pathway.These results may contribute to establish MDSCs induction system,which would provide a basis for the clinical application of MDSCs.

Effect of induction therapy regimens on monocytic myeloid-derived suppressor cells in kidney transplantation recipients / 中华器官移植杂志

Objective To investigate the effects of commonly used inductive agents on peripheral blood monocytic myeloid-derived suppressor cells (M-MDSCs) in renal transplantation recipients and to discuss their possible mechanism.Methods The enrolled patients received rabbit anti-thymocyte globulin (rATG) or basiliximab for induction therapy,with the maintenance immunosuppressive regimen of tacrolimus,mycophenolate mofetil and steroid.The number of CD11 b + CD33 + HLA-DR-CD14 + CD1 5-M-MDSCs and cytokine levels in peripheral blood,including interferon-γ(IFN-γ),interleukin-2 (IL-2),IL-4 and IL-6,were measured by flow cytometry before and 1 week,2 weeks,1 month,2 months,3 months after operation.Results A total of 47 recipients (29 given rATG 29,and 18 given basiliximab) were included in this study.Compared to the patients with basiliximab,asignificant increase in the frequency of M-MDSCs was observed in the rATG group at 2nd month after operation (5.5％ ± 2.8％ vs.3.8％ ± 1.6％,P＜0.001) and at 3rd month after operation (7.0 ％ ± 3.1％vs.4.1％ ± 2.3 ％,P＜ 0.001),while there was no significant difference in the cell number between the two groups.In the cytokine detection,levels of IL-2 and IL-4 in the rATG-treated recipients were significantly higher at 2nd weekpostoperation (Pr2 =0.032,and PIL-4 =0.019)and 1st month postoperation (PIL-2 =0.024,PIL-4 ＜0.001) than the basiliximab group.Conclusions ATG promotes the expansion of M-MDSCs,which is associated with the secretion of IL-2 and IL-4 due to the lymphocytes depletion.The synergistic immunosuppressive effect may contribute to the induction of immune tolerance.

Sleep disorders and its related risk factors in patients undergoing chronic peritoneal dialysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The prevalence of sleep disorders has been shown to be high in patients with chronic dialysis patients and may contribute to impaired quality of life and higher mortality in this population. However, there are few data on the relationship of sleep disorders and their risk factors in chronic dialysis patients. The aim of this study was to evaluate the relationship of sleep disorders and their risk factors in chronic dialysis patients.</p><p><b>METHODS</b>A total of 42 continuous ambulatory peritoneal dialysis (CAPD) patients were involved in this cross-sectional study. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI). Restless legs syndrome (RLS) was diagnosed according to the criteria of the International Restless Legs Syndrome Study Group. And depression was assessed by Hamilton depression scale. General information and laboratory data were collected.</p><p><b>RESULTS</b>The prevalence of sleep disorders was 47.6% in the CAPD patients. According to the PSQI, the 42 CAPD patients were divided into sleep disturbance group and non-sleep disorders group. There were no significant differences in age, gender, dialysis duration, hemoglobin, serum creatinine, urea nitrogen, β2-microglobulin, parathyroid hormone, calcium, and phosphorus between CAPD patients with sleep disorders and those without sleep disorders. But the level of serum albumin (Alb) in CAPD patients with sleep disorders was significantly lower than that in CAPD patients without sleep disorders (31.3 ± 1.4 vs. 34.3 ± 3.7, t = 3.603, P = 0.001) . And the prevalence of RLS and depression was significantly higher than that in CAPD patients without sleep disorders (RLS: 11/22 vs. 1/20, χ(2) = 10.395, P = 0.001; depression: 7/22 vs. 1/20, χ(2) = 4.886, P = 0.027). In CAPD patients with RLS, the prevalence of sleep disorders was significantly higher than that in CAPD patients without RLS (11/22 vs. 11/30, χ(2) = 10.395, P = 0.001). And in CAPD patients with depression, the prevalence of sleep disorders was significantly higher than that in CAPD patients without depression (7/8 vs. 15/34, χ(2) = 4.886, P = 0.027). In CAPD patients, bivariate correlation analysis showed that sleep disorders was negatively correlated with serum Alb (r = -0.606, P = 0.000) and positively correlated with RLS (r = 0.497, P = 0.001) and depression (r = 0.341, P = 0.029). Multivariate regression analysis revealed that the odds ratio of RLS, depression, and low serum Alb was 22.900, 42.209, and 0.597, respectively.</p><p><b>CONCLUSIONS</b>The prevalence of sleep disorders was relatively high in CAPD patients. RLS, depression, and low serum Alb were the risk factors for CAPD patients with sleep disorders.</p>

Identification of nPKCε-interacted proteins in the cortex of hypoxic preconditioned mice / 基础医学与临床

Objective Identify novel protein kinase Cε(nPKCε)-interacted proteins in the cortex of hypoxic preconditioned mice.Methods Immunoprecipitation (IP) and two-dimensional electrophoresis (2-DE) combining with ImageMaster 2D Platinum software were applied to analyze the differential expressions of nPKCe-interacted proteins;the target protein spots were identified by matrix-associated laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and Western blot.Results Compared with control group,there were 34 upregulated protein spots and 20 downregulated protein spots in cytosolic fraction,while 27 upregulated prtein spots and 28 downregulated protein spots were determined in particulate fraction of cerebral cortex of HPC mice.The levels of nPKCε-interacted HSP 70 and 14-3-3γ/protein expressions increased significantly in both cytosolic and particulate fractions;but the protein level of nPKCε-interacted HSP60 increased only in particulate fraction of cerebral cortex of HPC mice.Conclusion nPKCε might be involved in the development of cerebral HPC via the regulation of its interacted proteins such as HSP60,HSP70 and 14-3-3γ.

CRMP-2 is involved in hypoxic preconditioning-induced neuroprotection against cerebral ischemic injuries of mice / 基础医学与临床

Objective To investigate whether conventional protein kinase C (cPKC ) βⅡ-interacting collapsin response mediator protein-2 (CRMP-2) provides neuroprotection against cerebral ischemic (I) injuries. Methods Male BALB/c mice were randomly divided into normoxic control (Nor) , HPC, Nor + Sham, HPC + Sham, Nor + I and HPC + I groups (n = 6 per group). Using our HPC and MCAO mouse models, we applied immunoprecipita-tion, two-dimensional electrophoresis and mass spectrometry to characterize cPKCβⅡ-interacting proteins and combined with SDS-PAGE and Western blot to quantitatively analyze CRMP-2 phosphorylation and degradation levels in the brain of mice after HPC and MCAO. Results The expression level of 10 cPKCβⅡ-interacting proteins changed obviously in cerebral cortex of HPC mice when compared with Nor group. One of these proteins, CRMP-2 protein level increased in particulate fraction and decreased in cytosolic fraction of cerebral cortex of HPC mice. CRMP-2 phosphorylation level in ischemic core (Ic) of cerebral cortex decreased significantly ( P < 0. 05 , n = 6) as compared with that of Nor + sham group, but CRMP-2 phosphorylation level in HPC +I group increased significantly as compared with that of Nor +I group ( P < 0. 05, n = 6). In ischemic cortex, CRMP-2 degradation (proteolysis) was observed as the appearance of 55 ku breakdown products (BDP). However, the CRMP-2 degradation level, BDPs products decreased significantly in penumbra ( P) of ischemic cortex from HPC +I group when we compared with that of Nor +I group (P < 0. 05, n = 6 ). Conclusion CRMP-2 is involved in attenuating the decrease of CRMP-2 phosphorylation in ischemic core and in inhibiting its degradation in penumbra of cerebral cortex of mice thereby to lessen the ischemic injuries.

Troubleshooting experience of fridge used in medicine / 中国医疗器械杂志

This paper gives a brief introduction on common troubleshooting of fridge used in medicine and the corrective way of maintenance.