1.Regulation of mesenchymal stem cells on CD4+ Foxp3+ T cells in systemic lupus erythematosus
Shengnan ZHAO ; Huayong ZHANG ; Zhifeng GU ; Ting XU ; Ouyang JIN ; Lingyun SUN
Chinese Journal of Rheumatology 2008;12(10):663-666
Objective To investigate the in vivo or in vitro immune regulatory effects of allogeneic bone-marrow mesenchymal stem ceils (MSC) and human umbilical cord MSC on CD4+ Foxp3+ T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and in the spleen of MRL/Ipr mice. Methods Human MSC were isolated and expanded from bone marrow cells of healthy donors and infused into five SLE patients. The percentages of CD4+ Foxp3+ T cells in peripheral blood were detected by flow cytometry. Human peripheral blood mononuclear cells (PBMC) were prepared by centrifugation on a Ficoll Hypaque density gradient. The MSC and PBMC from unrelated donors (MSC:PBMC =1:1,1:10,1:50) were added into 24-well plates. After 72h of co-culture, the percentages of CD4+ Foxp3+ T cells were detected by flow cytome- try. Twenty four 18-week-old MRL/Ipr female mice were divided into 3 groups and were injected with umbilical cord MSC (1×106 cells for one time, 1×106 cells for three times and 0.5 ml sodium chloride as control respectively). The percentages of CD4+ Foxp3+ T cells in spleen and lymphoid nodes were detected by flow cytometry. Results The percentages of blood CD4+ Foxp3+ T cells at one week (4.8± 1.6)% and at three months (6.0±2.6)% post MSC transplantation for patients with SLE were both higher than that before transplantation (2.1±1.2)% (n=5,P<0.05). The co-culture of normal bone marrow MSC with PBMC from SLE patients resulted in a statistically significant increase of CD+ Foxp3+ T cells percentage in PBMC on a dose dependent manner (P<0.05). The percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients co-cultured with lupus MSC were lower than that of normal MSC (P<0.05). The cultured supematant of normal MSC also upregulated the percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients (P<0.05). The MRL/lpr mice that had been injected umbilical cord MSC for one time and three times had higher percentages of CD4+ Foxp3+T cells in the spleen but lower in the lymphoid nodes as compared with controls (P<0.01), but without statistical significant difference. Conclusion Allogeneic or heterogeneie MSC transplantation upregulates the percentages of CD4+ Foxp3+ T cells in SLE patients or in MRL/Ipr mice. Upregnlation of Treg population may be one of the mechanisms of MSC transplantation that is effective for SLE treatment.
2.The effect of hydroxychloroquine treatment on MRL/lpr lupus mice
Ouyang JIN ; Shengnan ZHAO ; Ting XU ; Zhifeng GU ; Huayong ZHANG ; Hong WANG ; Lingyun SUN
Chinese Journal of Rheumatology 2009;13(1):16-19,后插二
Objective To investigate the therapeutic effects and mechanisms of hydroxychloroquine (HCQ) in the MRL/lpr mice. Methods MRL/lpr mice were divided into HCQ, the artesunate (ART) and proteinuria was detected with Coomassi Brilliant blue method. Enzyme linked immunosorbent assay (ELISA) was used to measure the anti-doubM-stranded DNA (ds-DNA) antibody. Renal tissue sections were dyed By PAS methods. The percentage of CD4+ Foxp3+ T cells in the spleen and lymph nodes were detected by flow 2.0) mg groups were decreased than in the control group (4.8±3.2) mg (P<0.05). And it was also lower in the HCQ (2.8±1.1) mg and ART (2.4±1.9) mg group than in the control group (6.4±1.9) mg (P<0.01) at 30 in the control group (37.1±1.0) g (P<0.01), while serum creatinine decreased significantly (7.8±4.0) μmol/L than in the control group (12.5±2.3) μmol/L (P<0.05), and the serum anti ds-DNA antibodies levels (3047±renal damage in the HCQ group and in the ART group was Both significantly improved than that in the entages of CD4+ Foxp3+ T cells in spleen when compared with the control group (1.5±0.5)% (P<0.05). The mice in the HCQ group (0.68±0.33)% and in the ART group (0.97±0.28)% had higher percentages of CD4+ Foxp3+ T cells in lymph nodes as compared with control group (2.15±0.72)%(P<0.01). Conclusion HCQ is effective in treating MRL/lpr lupus mice. It can improve the pathologic lesions of lupus nephritis, reduce proteinuria and antibody production. Both HCQ and ART can up-regulate the percentage of CD4+ Foxp3+ T cells in spleen of MRL/lpr mice.
3.Characterization of the ultrastructure and cytoskeleton of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus
Zhifeng GU ; Shengnan ZHAO ; Huayong ZHANG ; Ting XU ; Ouyang JIN ; Kangxing ZHOU ; Haiou LIU ; Tianyi ZHANG ; Lingyun SUN
Chinese Journal of Rheumatology 2008;12(6):397-400,插3
Objective To explore ultrastructure and cytoskeleton characteristics of bone marrow-deftved mesenchymal stem cells (MSCs) in patients with systemic lupus erythematosus (SLE).Methods MSCs were isolated from bone marrow of 2 SLE patients and 2 healthy controls.Their ultrastrnctures were examined by transmission electron microscope (TEM).The expression pattern of actin and vinculin was assessed by laser confocal microscopy (LCM).Results MSCs in patients with SLE presented with signs of ageing and lots of autophagosome could be found in most of the cells.F-actin was aggregated and condensed at the:border of cytoplasm.Vinculin was arranged disorderly and condensed in the cytoplasm.Conclusion The change of uhrastructure and cytoskeleton patterns of bone marrow derived mesenchymal cells of SLE patients may play an important role in the abnormal proliferation of these cells in vitro.
4.Value of 18F-FDG PET/CT combined with conventional imaging modalities in TNM staging of rectal cancer
Changhong LI ; Zhen LIU ; Cheng GUO ; Huayong GU ; Xipeng ZHAO ; Lin GAO ; Xinglong GUO ; Yue LU ; Yanli WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(6):349-354
Objective:To investigate the value of 18F-FDG PET/CT combined with conventional imaging modalities in the evaluation of the depth of tumor invasion, regional lymph node metastasis, distant organ and lymph node metastasis (TNM staging), and the adjacent structure invasion of rectal cancer. Methods:Fifty-four patients (28 males, 26 females, age (65.8±11.0) years) with pathologically confirmed rectal cancer admitted to the Affiliated Qingdao Central Hospital of Qingdao University between September 2019 and June 2021 were retrospectively analyzed. 18F-FDG PET/CT examination, conventional imaging modalities including high-resolution MRI (HR-MRI), chest CT plain scan, upper abdominal MRI or CT plain scan+ enhanced examination were performed within 2 weeks before or after the rectal cancer being confirmed. The TNM staging and adjacent structural invasions including circumferential resection margin (CRM), extramural vascular invasion (EMVI), anal sphincter complex involvement were evaluated by 18F-FDG PET/CT and conventional imaging modalities separately or in combination, and those results based on imaging were compared with the pathological results or clinical follow-up results. χ2 test was used to compare the differences of diagnostic sensitivity, specificity and accuracy between the 18F-FDG PET/CT or conventional imaging modalities and combined examination. Results:The accuracy for T staging and the sensitivity and accuracy for N staging of the combined examination were 96.30%(52/54), 98.65%(73/74) and 93.91%(185/197), respectively, which were significantly higher than those of 18F-FDG PET/CT (85.19%(46/54), 66.22%(49/74), 81.73%(161/197); χ2 values: 3.97, 26.88, 13.66, all P<0.05). The specificity (91.06%, 112/123) and accuracy of the combined examination for N staging were higher than those of the conventional imaging modalities (77.24%(95/123), 83.76%(165/197); χ2 values: 8.81, 10.23, both P<0.05). The sensitivity and accuracy of the combined examination for M staging were higher than those of the conventional imaging modalities (97.01%(65/67) vs 73.13%(49/67), 95.95%(71/74) vs 68.92%(51/74); χ2 values: 15.05, 18.66, both P<0.001). The sensitivities of the combined examination in evaluating CRM and EMVI were 100%(22/22) and 95.00%(19/20), and the accuracies were 98.15%(53/54) and 96.30%(52/54), all of which were higher than those of 18F-FDG PET/CT (CRM: 54.55%(12/22), 74.07%(40/54); EVMI: 30.00%(6/20), 74.07%(40/54); χ2 values: 12.94, 13.08, 18.03, 10.56, all P<0.01). The accuracy of the combined examination in evaluating EMVI was higher than that of the conventional imaging modalities (85.19%(46/54); χ2=3.97, P=0.046). Conclusion:18F-FDG PET/CT combined with conventional imaging modalities can improve the diagnostic efficacy for TNM staging and assessment of adjacent structural invasion in rectal cancer.