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Objective:To explore the association of bone resorption marker β carboxyterminal peptide of collagen Ⅰ (β-CTX) with hypercalcemia in patients with Graves′ disease (GD).Methods:287 patients with GD who were hospitalized in the endocrinology department of Fuyang People′s Hospital from January 2021 to December 2021 were divided into control group ( n=251) and hypercalcemia group ( n=36) according to the corrected blood calcium level. The clinical data and serum β-CTX level of the two groups were compared. Logistic regression model was used to analyze the risk factors of hypercalcemia in GD patients. Pearson correlation was used to analyze the correlation between serum β-CTX level and other indexes. Results:Of the 287 GD patients, 36 were diagnosed as hypercalcemia, and the incidence of hypercalcemia was 12.54%. The levels of free triiodothyronine (FT3), free thyroxine (FT4), blood phosphorus (P) and β-CTX in hypercalcemia group were higher than those in control group, and the total parathyroid hormone (iPTH) in hypercalcemia group were lower than those in control group (all P<0.05). Multivariate Logistic regression analysis showed that FT3 ( OR=1.283, 95% CI: 1.049-1.570, P<0.05), iPTH ( OR=0.924, 95% CI: 0.863-0.989, P<0.05), β-CTX ( OR=2.488, 95% CI: 1.193-5.189, P<0.05) were the influencing factors for hypercalcemia in GD patients. Pearson correlation analysis showed that β-CTX was positively correlated with FT3, FT4, blood calcium, P, alkaline phosphatase (ALP), total procollagen type I amino end terminal peptide (PINP), N-bone-gamma-carboxyglutamic-acid-containing proteins (N-MID) and 25(OH)D, and negatively correlated with iPTH (all P<0.05). Conclusions:β-CTX is highly expressed in the serum of GD patients with hypercalcemia, which is a risk factor for the occurrence of hypercalcemia in GD patients.
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Objective To study the alterations of CD19+CD5+CD1dhigh B, Th1 and Th17 cells in non-obese diabetic ( NOD) mice and the correlation between B10 cells and type 1 diabetes in NOD mice. Methods Flow cytometry ( FCM) was used to measure the levels of CD19+CD5+CD1dhigh B, CD19+IL-10+B, CD4+IFN-γ+Th1, CD4+IL-17+Th17 and CD4+CD25+Foxp3+T cells in NOD mice ( 4 weeks old NOD mice:group A, n=10;8 weeks old NOD mice:group B, n=10; NOD mice with diabetes: group C, n=10) and age-matched C57BL/6 mice ( control group, n=20 ) .Hematoxylin-eosin staining of pancreatic tissues was performed for histopathological assessment of the development of insulitis in NOD mice.Results (1) Histopathological analysis showed that mice from A, B and C groups respectively showed no insulitis, insulitis and obvious insulitis with no intact islets.(2) The highest levels of B10 cells in NOD mice were ob-served in group B, followed by those in group C and group A (P0.05).More B10 cells were detected in pancreatic lymph nodes than in other tissues of 8 weeks old NOD mice, the levels of which were also higher than those in pancreatic lymph nodes of mice form group C ( P0.05).Conclusion The percentages and distribution of B10 cells in NOD mice changed with age and the development of insulitis.The decrease of B10 cells might participate in the development of type 1 diabe-tes in NOD mice.
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Objective To investigate the phenotypes and percentages of B 10 cells in different tis-sues from wild-type mice and to identify their biological functions .Methods The percentages of B10 cells derived from different tissues of mice and their responses to lipopolysaccharide ( LPS) stimulation were ana-lyzed by flow cytometry .Magnetic-activated cell sorting ( MACS ) and fluorescence-activated cell sorting (FACS) were used to purify B10 cells, CD4+CD25-T cells and Treg cells.CD4+CD25-T cells and Treg cells labeled by CFSE were co-cultured with or without B10 cells, and then their proliferation were evaluated after 72 h.Results (1) A subset of CD19+CD5+CD1dhigh regulatory B cells was identified in spleen , pe-ripheral blood and lymph nodes from wild-type mice , of which the highest frequency was detected in spleen (3.95%±0.79%, P<0.05).The isolated B cells from different tissues were stimulated by LPS , PMA, ionomycin and monensin (L+PIM) in vitro to express IL-10.Among them, splenic CD19+IL-10+B cells showed the highest expression of IL-10 (P<0.05).(2) Prolonged LPS stimulation (48 h) to CD5+CD1dhigh B cells enhanced the expressions of IL-10 (P<0.01).(3) CD19+CD5+CD1dhigh B cells inhibited the prolif-eration of CD4+CD25-T cells in vitro in a dose-dependent manner (P<0.01), but increased the secretion of IL-10 by CD4+T cells (P<0.01) and the proliferation of Treg cells in vitro (P<0.01).Conclusion Com-pared with other tissues , the percentage of B10 cell subset in spleen is the highest in wild-type mouse , and B10 cells subset can be activated through Toll-like receptor ( TLR ) signaling pathway .The responses of CD4+CD25-T cells and Treg cells in co-culture with B10 cells are regulated by B 10 cell subset through an increased IL-10 production .B10 cells might be a useful cell population for the treatment of inflammatory au-toimmune diseases.