白色脂肪组织(WAT)在调节全身代谢稳态和代谢性心血管疾病发生、进展中发挥重要作用.WAT来源的细胞外囊泡(EV)的产生异常和功能障碍与多种代谢性心血管疾病的发病有关.EV通过影响内皮功能、炎症反应、血管钙化、胰岛素抵抗和脂质代谢等途径参与动脉粥样硬化的发生发展,还通过影响糖脂代谢、氧化应激和钙超载等机制参与心肌缺血再灌注损伤,并可通过调节miRNA参与心力衰竭的发病过程.WAT来源EV产生异常和功能障碍的机制及其携带miRNA影响代谢性心血管疾病的机制仍有待进一步研究.

目的研究糖蛋白M6B（Glycoprotein M6B,GPM6B）在白色脂肪细胞向棕色脂肪表型转变中的作用。方法 （1） Western blot和qPCR检测小鼠白色脂肪组织（white adipose tissue, WAT）和棕色脂肪组织（brown adipose tissue, BAT）中GPM6B的表达。（2）分别用对照组（shScramble）和敲低GPM6B组（shGPM6B1和shGPM6B2）的慢病毒液感染C3H10T1/2小鼠间充质细胞系,荧光显微镜下观察病毒的转染效率。Western blot和qPCR检测GPM6B的干涉效率。（3）将shScramble, shGPM6B1和shGPM6B2的稳转细胞系分别给予白色脂肪诱导分化液8 d,油红O染色观察脂滴形态,Western blot检测棕色脂肪标记基因解偶联蛋白（uncoupling protein, UCP1）的表达,qPCR检测热生成相关基因的mRNA表达。结果 （1）与WAT比,BAT中GPM6B的表达在蛋白和mRNA水平均显著降低（P＜0.05）。（2）与shScramble组相比,shGPM6B1组和shGPM6B2组GPM6B的干涉效率在蛋白和mRNA水平均显著降低（P＜0.05）。（3）敲低GPM6B后,白色脂肪细胞的脂滴变小,UCP1的表达和热生成相关基因的mRNA均显著升高（P＜0.05）。结论敲低GPM6B促进白色脂肪细胞向棕色脂肪表型转变。

Objective To investigate the alterations of plasmacytoid dendritic cells ( pDC) in pa-tients with primary Sj?gren′s syndrome ( pSS) and their significance.Methods Peripheral blood samples were collected from 39 patients with pSS and 15 healthy subjects.The percentages of pDC ( Lin-CD123+HLA-DR+) in peripheral blood samples were analyzed by flow cytometry.The levels of IFN-α, TNF-αand IL-6 in serum samples were detected by ELISA.Results The percentages of pDC in patients with pSS were lower than those in healthy subjects (P<0.05).Higher levels of pDC were detected in patients positive for anti-Sj?gren syndrome antigen A ( anti-SSA) antibody as compared with those in patients negative for anti-SSA antibody.The levels of TNF-αand IL-6 in serum samples from patients with pSS were higher than those in healthy subjects.Conclusion Patients with pSS showed decreased percentages of pDC population, but increased levels of TNF-αand IL-6.The changes of pDC population, TNF-αand IL-6 in peripheral blood might play active roles in the pathogenesis of pSS.

To investigate the impact of hydroxychloroquine(HCQ) on plasmacytoid dendritic cells activation induced by CpG-ODN in vitro,cord blood(250-300 ml) was collected from delivery room.Density gradient centrifugation was used to get the cord blood monocytes and BDCA-4 beads sorting was used to obtain the precursor pDCs.Precursor pDCs was co-cultured with IL-3,IL-3 plus CpG-ODN2216,and IL-3 plus CpG-ODN2216+HCQ,respectively for 3 days.Then flow cytometry was used to measure the pDCs surface expression of BDCA-2,CD86 and CD32;while ELISA was employed to detect the concentration of IFN-α in the supernatants.We found that CpG-ODN2216 activated pDCs demonstrated higher expressions of CD86 and CD32,more IFN-α production,but lower expression of BDCA-2 in vitro(P 0.05).And HCQ could significantly down-regulate the CD32 expression,CD32 MFI level,and IFN-α level in the supernatants(P 0.05).In conclusion,HCQ may inhibit pDCs activation and IFN-α secretion induced by CpG-ODN2216 via reducing CD32 expression on pDCs.

<正>抗疟药主要指喹啉类化合物和吖啶化合物,前者如羟氯喹、氯喹、奎宁、阿莫地喹、喷他喹、喹啉衍生物(4-氨喹啉衍生物、8-氨喹啉衍生物),后者如米帕林。抗疟药最初用来治疗疟疾,20世纪50年代,抗疟药被誉为免疫调节剂,广泛用于系统性红斑

Background Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by immunological intolerance, hyperactivity of various immunocyte and production of a variety of autoantibodies. Although etiology of the disease remains unclear, there is an increasing evidence that the presence and accumulation of dendritic cells in target tissues may play a role in the pathogenesis of SLE. Objectives To explore the changes and significance of plasmacytoid dendritic cells (PDC), myeloid dendritic cells (MDC) and cytokines in peripheral blood in patients with SLE. Methods The propotions of PDC and MDC in peripheral blood from 38 SLE and 15 normal individuals were measured by flowcytometer. The concentrations of IFN-α, TNF-α and IL- 6 were detected using ELISA. Results 1. The levels of PDC and MDC in peripheral blood of SLE patients decreased significantly comparing with normal controls (P<0.01). 2. It was found that the percentage of PDC in SLE patients with stable disease was significantly higher than patient with active disease (P<0.05). No significant difference was found in MDC between patients with and without disease activity. 3. SLE patients with proteinuria had lower percentage of PDC comparing with patients without proteinuria (P<0.05). In addition, anti-dsDNA negative patients had higher percentage of PDC than anti-dsDNA positive patients (P<0.05). 4. The percentage of PDC was correlated significantly with SLEDAI, C3 and ESR (with r value of -0.363, 0.451, -0.453 and p value of 0.031, 0.001, 0.008 respectively). 5. Further study found SLE patients had higher level of IFN-α, IL-6 and TNF-α than controls (P<0.05). No correlation was found between the percentage of PDC, MDC and level of IFN-α, IL-6 and TNF-α. Conclusions The percentages of PDC and MDC in peripheral blood are decreased in patients with SLE. The reduction of PDC is correlated with disease activity and kidney damage of SLE. Disclosure of Interest None Declared

Objective To invesigate the clinical significance of anti-cyclic citrullinated peptides antibodies(anti-CCP) in systemic lupus erythematosus(SLE) patients with arthritis,and to explore the association of anti-CCP with erosive arthritis and judge prognosis in SLE patients.Methods 145 SLE patients were examined for anti-CCP and IgM-RF with enzyme-linked immunosorbent assay method and rate turbidimetric method.Results The frequency of ant-CCP was 24.1% in this group of SLE patients.The frequency of arthritis was significantly higher in SLE patients with anti-CCP than without anti-CCP(77.1% VS 38.2%,P<0.05).Anti-CCP antibodies were more frequently detected in SLE patients with arthritis than those without arthritis(39.1% VS 10.5%,P<0.05).The frequency of RF-positive group with Anti-CCP than that without anti-CCP was(38.5% VS 24.4%,P<0.05).When compared with anti-CCP negative patient,erosive arthritis occurred more often in anti-CCP positive patients than nagative anti-CCP patients(25.7% VS 4.6%,P<0.001).Conclusion Anti-CCP antibodies have a prevalence in our group of SLE patients and its presence is closely associated with erosion of bone and joint destruction.

Objective To explore the role of hydroxychloroquine (HCQ) in ultraviolet B (UVB)- induced expression of interleukin (IL)-10 and interferon (IFN)-γ from CD4+T cells in patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 healthy controls were enrolled in the study. CD4+ T cells were isolated using magnetic beads from SLE patients and healthy controls. HCQ was added in culture media before and after irradiation with UVB 311 nm narrow band ultraviolet B (NB-UVB). The levels of IL-10 and IFN-γ in the supernatant were detected with enzyme-linked immunosorbent (ELISA). Comparisons between groups were performed by t-test. Results The level of IL-10 was higher in SLE patients [(27±4) pg/ml] than that in healthy controls [(18±3) pg/ml, P=0.011]. After exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages, the level of IL-10 was increased significantly in patients with active disease (P=0.022, P=0.048). After exposure of CD4+T cells to UVB in 100 mJ/cm2 dosages, the levels of IL-10 was higher in patients with active disease [(77±42) pg/ml] than patients with stable disease [(24± 4) pg/ml, P=0.029]. When CD4+ T cell were cultured with HCQ, IL-10 and IFN-γ levels in patients with active disease [(2.6±4.0), (17.5±2.3) pg/ml] were decreased significantly (P=0.018, P=0.017). HCQ reversed UVB-induced IL-10 expression in active SLE patients after exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages (P=0.037, P=0.04). HCQ also reversed UVB-induced IFN-7 expression in active SLE patients and stable SLE patients after exposure to CD4+T cells with UVB in 100 mJ/cm2 dosages (P=0.013, P= 0.049). Conclusion UVB can aggravate the imbalance of Th1 and Th2 cytokines. HCQ inhibits UVB-induced IL-10 and IFN-7 expression of CD4+T cells in patients with SLE, especially in patients with active disease.

Objective To explore the effect of hydroxychloroquine on ultraviolet ray-induced genomic DNA hypomethylation in CD4+ T cells from patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 normal human controls were enrolled in the study. CD4+ T cells were isolated from these subjects by using magnetic beads, and cultured. Hydroxychloroquine of 50 mg/L was added to the culture medium of CD4+ T cells before or after the exposure to narrow band ultraviolet B (NB-UVB) 311 nm.After additional culture, the levels of genomic DNA methylation in CD4+ T cells were determined with the Imprint Methylated DNA Quantification kit. Results The levels of DNA methylation was lower in SLE patients than in the normal controls [(3.922 ±2.215)% vs. (10.210 ± 5.573)%, t= 3.450, P = 0.026]. After exposure to UVB at 45 and 100 mJ/cm2, the DNA methylation level in patients with active SLE decreased from (7.235 ±3.846)% to (1.784 ± 1.033)% and (1.932 ± 1.844)% respectively (t= 3.000, 4.118, both P< 0.05). Decreased DNA methylation level was observed in CD4+ T cells from patients with active SLE compared with those from patients with stable SLE and normal human controls [(1.932 ± 1.844)% vs. (7.235 ± 3.846)% and (5.472 ±5.573)%, t = 2.648, 3.000, both P< 0.05] after irradiation with UVB of 100 mj/cm2. A significant increase in the methylation level was observed in active SLE patient-derived CD4+ T cells treated with hydroxychloroquine following the irradiation with UVB of 45 (4.698% ± 1.948%) and 100 mJ/cm2(8.698% ± 3.151%) compared with those only treated with UVB irradiation (t = 4.827, 3.184, both P< 0.05), as well as in those treated with hydroxychloroquine before and after the irradiation with UVB of 45 mJ/cm2 compared with those receiving irradiation alone [(5.404 ± 2.308)% vs. (1.784 ± 1.033)%, t = 4.827, P< 0.01]. Conclusion Hydroxychloroquine can reverse the UVB-induced genomic DNA hypomethylation in CD4+ T cells from patients with SLE,especially in those from patients with active SLE.