Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)％ compared to the gld+/+ apoE+/+ group [(0.039±0.005)％,P＜0.01],the gld group [(0.025±0.001)％,P＜0.05],and the apoE-/-group [(0.028±0.002)％,P＜0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)％ compared to the gld+/+apoE+/+ group [(0.42±0.05)％,P＜0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)％] compared to the gld+/+apoE+/+ group [(87.5±3.0)％,P＜0.01],and the gld group [(84.2±6.0)％,P＜0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)％] compared to the gld+/+ apoE+/+ group [(86.0±7.3)％,P＜0.01],the gld group [(73.0±1.0)％,P＜0.01],and the apoE-/-group [(65.0±4.0)％,P＜0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P＜0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.

Objective To investigate the changes and clinical significance of γδT cells and their major functional subsets γδT1 and γ8T17 cells in the peripheral blood of patients with systemic lupus erythematosus(SLE).Methods Ratios and absolute numbers of γδT cells,γδT1 cells and γT17 cells in peripheral blood were detected by flow cytometry in 21 SLE patients(SLE)group and 16 healthy controls(HC group).Correlations of γδT cells,γδT1 cells and γ8T17 cells in the peripheral blood with clinical indicators were analyzed by Pearson correlation analysis.Results In SLE pa-tients,the percentage of γδT cells to lymphocytes in the peripheral blood was(2.30±1.10)％,which was significantly lower than(3.30±1.51)％in the HC group(P＜0.05);the percentage ofγδT1 cells to lymphocytes in peripheral blood was(0.93±0.67)％,which was significantly lower than(2.09±1.21)％in the HC group(P＜0.001);the ratio of γδT17 cells to lymphocytes in the peripheral blood was(1.53±2.82)‰,which showed an increasing trend when compared to(0.18± 0.12)％o in the HC group(P＞0.05).The absolute number of γδT cells in the peripheral blood in SLE patients was(2.88±2.18)× 104/mL,which was significantly lower than(7.96±3.96)× 104/mL in the HC group(P＜0.000 1);the absolute number of γδT1 cells in the peripheral blood in SLE patients was(1.20±1.17)× 104/mL,which was significantly lower than(5.05±3.04)× 104/mL in the HC group(P＜0.000 1);the absolute number of γδT17 cells in the peripheral blood in SLE patients was(1.03±1.08)× 103/mL,which was significantly higher than(0.40±0.24)×103/mL in the HC group(P＜0.05).The proportion of γδT17 cells in the peripheral blood of SLE pa-tients was positively correlated with the Systemic Lupus Erythematosus Disease Activity Index(SLE-DAI)(r=0.59,P＜0.01)and erythrocyte sedimentation rate(r=0.65,P＜0.01),while the absolute number of y8T17 cells was positively correlated with the SLEDAI score(r=0.59,P＜0.01)and negatively correlated with the complement C3 level(r=-0.53,P＜0.05).Conclusion The significant decrease in the number of γδT cells in the peripheral blood of SLE patients is mainly due to the reduction of its γδT1 cell subset;γδT17 cells play an important role in the patho-genesis and activity of SLE.

Objective To investigate the changes and clinical significance of γδT cells and their major functional subsets γδT1 and γ8T17 cells in the peripheral blood of patients with systemic lupus erythematosus(SLE).Methods Ratios and absolute numbers of γδT cells,γδT1 cells and γT17 cells in peripheral blood were detected by flow cytometry in 21 SLE patients(SLE)group and 16 healthy controls(HC group).Correlations of γδT cells,γδT1 cells and γ8T17 cells in the peripheral blood with clinical indicators were analyzed by Pearson correlation analysis.Results In SLE pa-tients,the percentage of γδT cells to lymphocytes in the peripheral blood was(2.30±1.10)％,which was significantly lower than(3.30±1.51)％in the HC group(P＜0.05);the percentage ofγδT1 cells to lymphocytes in peripheral blood was(0.93±0.67)％,which was significantly lower than(2.09±1.21)％in the HC group(P＜0.001);the ratio of γδT17 cells to lymphocytes in the peripheral blood was(1.53±2.82)‰,which showed an increasing trend when compared to(0.18± 0.12)％o in the HC group(P＞0.05).The absolute number of γδT cells in the peripheral blood in SLE patients was(2.88±2.18)× 104/mL,which was significantly lower than(7.96±3.96)× 104/mL in the HC group(P＜0.000 1);the absolute number of γδT1 cells in the peripheral blood in SLE patients was(1.20±1.17)× 104/mL,which was significantly lower than(5.05±3.04)× 104/mL in the HC group(P＜0.000 1);the absolute number of γδT17 cells in the peripheral blood in SLE patients was(1.03±1.08)× 103/mL,which was significantly higher than(0.40±0.24)×103/mL in the HC group(P＜0.05).The proportion of γδT17 cells in the peripheral blood of SLE pa-tients was positively correlated with the Systemic Lupus Erythematosus Disease Activity Index(SLE-DAI)(r=0.59,P＜0.01)and erythrocyte sedimentation rate(r=0.65,P＜0.01),while the absolute number of y8T17 cells was positively correlated with the SLEDAI score(r=0.59,P＜0.01)and negatively correlated with the complement C3 level(r=-0.53,P＜0.05).Conclusion The significant decrease in the number of γδT cells in the peripheral blood of SLE patients is mainly due to the reduction of its γδT1 cell subset;γδT17 cells play an important role in the patho-genesis and activity of SLE.

Objective To explore the correlation of CPNE3 expression with long-term prognosis of patients with gastric cancer(GC)and the possible mechanism.Methods We retrospectively collected the data of 104 GC patients undergoing radical surgery in our hospital from February,2013 to October,2017.TCGA database and immunohistochemistry were used to analyze CPNE3 expression level in GC tissues and its effects on tumor progression and long-term prognosis of the patients.GO analysis was performed to predict the biological role of CPNE3 in GC.We also conducted cell experiments with MGC803 cells and observed the effects of CPNE3 knockdown,CPNE3 overexpression and LY294002(a PI3K/AKT inhibitor)treatment on cell apoptosis and cellular expressions of apoptotic proteins using flow cytometry and Western blotting.Results TCGA analysis and immunohistochemistry both showed high expressions of CPNE3 in GC(P<0.05).The patients with high CPNE3 expressions had a reduced 5-year survival(P<0.01),and a high CPNE3 expression,CEA level≥5 μg/L,CA19-9 level≥37 kU/L,T3-T4 stage,and N2-N3 stage were all independent risk factors for a lowered 5-year survival rate after surgery.The sensitivity and specificity of CPNE3 for predicting 5-year mortality was 79.59%and 74.55%,respectively(P<0.05).GO analysis predicted that CPNE3 negatively regulated GC cell apoptosis.In MGC803 cells,CPNE3 knockdown significantly increased cell apoptosis,enhanced Bax and Cleaved Caspase-3 expressions and decreased Bcl-2 expression,while CPNE3 overexpression produced the opposite results(P<0.05).The cellular expressions of p-PI3K and p-AKT were significantly decreased following CPNE3 knockdown and increased following CPNE3 overexpression(P<0.05).Treatment with LY294002 obviously attenuated the inhibitory effect of CPNE3 overexpression on apoptosis of MGC803 cells(P<0.05).Conclusion CPNE3 is highly expressed in GC tissues and affects the long-term prognosis of the patients possibly by inhibiting GC cell apoptosis through activation of PI3K/AKT signaling.

Objective To explore the correlation of CPNE3 expression with long-term prognosis of patients with gastric cancer(GC)and the possible mechanism.Methods We retrospectively collected the data of 104 GC patients undergoing radical surgery in our hospital from February,2013 to October,2017.TCGA database and immunohistochemistry were used to analyze CPNE3 expression level in GC tissues and its effects on tumor progression and long-term prognosis of the patients.GO analysis was performed to predict the biological role of CPNE3 in GC.We also conducted cell experiments with MGC803 cells and observed the effects of CPNE3 knockdown,CPNE3 overexpression and LY294002(a PI3K/AKT inhibitor)treatment on cell apoptosis and cellular expressions of apoptotic proteins using flow cytometry and Western blotting.Results TCGA analysis and immunohistochemistry both showed high expressions of CPNE3 in GC(P<0.05).The patients with high CPNE3 expressions had a reduced 5-year survival(P<0.01),and a high CPNE3 expression,CEA level≥5 μg/L,CA19-9 level≥37 kU/L,T3-T4 stage,and N2-N3 stage were all independent risk factors for a lowered 5-year survival rate after surgery.The sensitivity and specificity of CPNE3 for predicting 5-year mortality was 79.59%and 74.55%,respectively(P<0.05).GO analysis predicted that CPNE3 negatively regulated GC cell apoptosis.In MGC803 cells,CPNE3 knockdown significantly increased cell apoptosis,enhanced Bax and Cleaved Caspase-3 expressions and decreased Bcl-2 expression,while CPNE3 overexpression produced the opposite results(P<0.05).The cellular expressions of p-PI3K and p-AKT were significantly decreased following CPNE3 knockdown and increased following CPNE3 overexpression(P<0.05).Treatment with LY294002 obviously attenuated the inhibitory effect of CPNE3 overexpression on apoptosis of MGC803 cells(P<0.05).Conclusion CPNE3 is highly expressed in GC tissues and affects the long-term prognosis of the patients possibly by inhibiting GC cell apoptosis through activation of PI3K/AKT signaling.