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ABSTRACT: To investigate the correlation of the functional disequilibrium of regulatory T cells (Treg)/T-helper (Th17) cells with calcification and to explore the significance of their influence on the outcome of cardiovascular disease (CVD) in uremic patients after hemodialysis (HD).
Out of 66 uremia patients, 36 patients had CVD after HD (maintenance hemodialysis (MHD) group1) and 30 patients did not have CVD (MHD group2). Twenty healthy volunteers were selected as normal control group. Peripheral blood mononuclear cells were isolated and treated with recombinant human bone morphogenetic protein-2 (rhBMP-2). Treg and Th17 frequencies were measured by flow cytometry. Forkhead/winged helix transcription factor (Foxp3) and retinoic acid receptor-related orphan receptor-γt (ROR-γt) mRNA expressions were measured by real-time quantitative polymerase chain reaction. Levels of interleukin (IL)-10 and IL-17 were detected by enzyme-linked immunosorbent assay.
When compared with controls, rhBMP-2 upregulates Treg/Th17 functional disequilibrium in uremia patients, displaying higher Treg and Th17 frequencies, Foxp3 and ROR-γt expressions, and levels of cytokines (p < 0.05). These differences were also significant between MHD group1 and group2 (p < 0.05). It was also observed that Treg/Th17 functional disequilibrium was not only correlated with a calcification state but also consistent with the CVD.
The Treg/Th17 cell function disequilibrium might act synergistically with calcification in the high incidence of CVD after HD.
Renal Failure 04/2012; 34(6):697-702. · 0.82 Impact Factor
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ABSTRACT: We used high-resolution atomic force microscopy (AFM) to examine possible changes in the morphology of peripheral blood mononuclear cells (PBMCs), and to investigate their influence on vascular calcification in uremic patients on maintenance hemodialysis (MHD). 36 uremic patients had cardiovascular diseases after MHD (MHD group1) and 30 uremic patients did not (MHD group 2), and 20 healthy volunteers were the control group. The extent of coronary artery calcification was assessed with coronary artery calcification score (CACS). AFM was used to analyze PBMCs nuances. Concentrations of bone morphogenetic protein-2 (BMP-2) in PBMC supernatants were detected by ELISA. Protein expressions of BMP-2 were measured by Western blot. No significant differences in PBMC morphology were observed among groups by light microscopy. AFM images revealed that uremic patients exhibited significant differences of PBMC morphology and vascular calcification when compared with healthy volunteers. The PBMCs in uremic patients were larger in volume, mean height, half-maximum amplitude, average roughness and higher concentrations and expression of BMP-2 and CACS (P < 0.05), with granular processes or caveolae of uneven size distributed over cell surfaces. These differences were also significant between MHD group 1 and group 2 (P < 0.05). PBMC volume, mean height, half-maximum amplitude, and average roughness were positively correlated with BMP-2 and CACS. Moreover, the correlation PBMC with BMP-2 was higher than with CACS. PBMC morphology in MHD patients was related to the degree of vascular calcification. The larger mean height, half-maximum amplitude, average roughness and cell volume were, the higher degree of vascular calcification was.
Therapeutic apheresis and dialysis: official peer-reviewed journal of the International Society for Apheresis, the Japanese Society for Apheresis, the Japanese Society for Dialysis Therapy 04/2012; 16(2):173-80. · 1.39 Impact Factor
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ABSTRACT: Diabetic nephropathy (DN) is a major cause of type 2 diabetes mellitus (T2DM) mortality. Innate immunity has been shown to be closely associated with the occurrence and progression of T2DM-associated complications. In this study, we investigated the expression of Toll-like receptor 4 (TLR4) and CD14(+)CD16(+) monocytes in patients with T2DM and DN patients with uremia and TLR4 response to lipopolysaccharide (LPS), and to further explore the potential effects of inflammatory immune response in T2DM and DN uremia. Thirty DN patients with uremia, 28 T2DM patients, and 20 healthy volunteers were enrolled for the determination of CD14(+)CD16(+) fluorescence intensity and TLR4 expression on monocytes by using peripheral blood flow cytometry. Serum C-reactive protein (CRP) level was determined by using the immunoturbidimetry. Peripheral blood mononuclear cells (PBMCs) were isolated and stimulated with LPS for 24 h. monocytes were collected to detect NF-κB p65 and phosphorylated STAT5(p-STAT5) expressions by using Western blotting. Supernatants were sampled for the determination of interleukin-6 (IL-6) concentration by using ELISA. Compared to normal control, T2DM patients and DN uremic patients had a significantly higher CD14(+)CD16(+) fluorescence intensity, TLR4 expression, serum IL-6 and CRP level, whilst these biomarkers were more upregulated in DN uremic patients than in T2DM patients. Following the exposure to LPS, PBMCs showed a significant upregulation in NF-κB-p65 and p-STAT5 expression and a remarked increase in Supernatants IL-6 level, in a positive correlation with disease severity. Our results suggest that the disturbance in proinflammatory CD14(+)CD16(+) monocytes occurs in T2DM and DN uremic patients. Such immunological dysfunction may be related to the activation of TLR4/NF-κB and STAT5 signaling pathways underlying the immune abnormalities of CD14(+)CD16(+) monocytes.
Inflammation 08/2011; 35(1):388-96. · 1.75 Impact Factor
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ABSTRACT: The objective of this paper was to study the change of P38MAPK and Fas in the apoptosis of THP-1 cells induced by allicin.
The proliferation inhibition rates of THP-1 cells after various treatments were examined by MTT assay. Apoptosis rate was determined with Annexin V- FITC/PI double staining by flow cytometry. The expression and distribution change of the phosphorylation p38MAPK (P-p38MAPK) were detected by immunohistochemical staining. The changes of P-p38 MAPK and Fas proteins were detected by Western blot.
The proliferations of leukemia cell line THP-1 are inhibited by allicin. MTT assay showed that allicin can inhibit the proliferation of the THP-1 cell, and the inhibition was dependent on both dose and time. The IC50 of 72 hours was 12.8 mg x L(-1). Apoptosis rate detected by Annexin V-FITC/PI was proportional to the concentration of the allicin. After the immunohistochemical staining test, the P-p38MAPK was located in the cell nucleus and plasma, showing deep brown, when adding allicin to THP-1 cell. Western blot test showed that the P-p38MAPK proteins expression was proportional to the concentration of Allicin and was also dose dependent. The levels of P-p38MAPK in negative control group, 1/2 IC50 of 72 hours group and IC50 of 72 hours group were 0.259 8 +/- 0.013 2, 0.61 2 +/- 0.008 3 and 0.505 6 +/- 0.005 5 respectively, and the levels of Fas proteins were 0.287 4 +/- 0.008 9, 0.426 8 +/- 0.007 9 and 0.597 1 +/- 0.010 9 respectively. The difference was statistically significant when compared with the negative control group (P < 0.01).
Allicin can significantly induce THP-1 cells apoptosis, and its mechanism may be related to the activation of P38MAPK/Fas.
Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica 06/2009; 34(11):1439-43.
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ABSTRACT: To investigate the ability of alveolar macrophage (AM) and peripheral blood monocyte (PBMC) from lung cancer patients to produce Interleukin-10 (IL-10) and to evaluate the local and general cellular immune state of lung cancer patients.
AM and PBMC were obtained from 57 patients with lung cancer, 33 patients with benign pulmonary diseases and 12 healthy volunteers. IL-10 in the culture supernatants was measured quantitatively by ELISA.
(1) Elevated levels of IL-10 produced by PBMC were found in lung cancer group with respect to healthy volunteers and patients with benign pulmonary diseases (148.60±35.56 vs 93.83±9.22 and 108.91±15.95 ng×L⁻¹ respectively) (P < 0.001). The level of IL-10 by AM from lung cancer patients was 132.06±30.42 ng×L⁻¹, which was remarkably higher than that of healthy volunteers (92.67±11.22 ng×L⁻¹) and benign pulmonary diseases (94.39±10.04 ng×L⁻¹) (P < 0.001). (2) The level of IL-10 produced by PBMC from lung cancer group of stage IV was significantly higher than that of stage I+II (178.33±13.52 vs 131.57±25.35 ng×L⁻¹) (P < 0.001). AM from lung cancer patients of stage III and IV produced more IL-10 than that of stage I+II did (150.13±15.57 and 160.50± 18.75 vs 117.05±28.12 ng×L⁻¹ respectively) (P < 0.001); PBMC from small cell lung cancer patients produced higher level of IL-10 than squamous cell carcinoma and adenocarcinoma did (194.83±23.88 vs 140.37± 27.00 ng×L⁻¹ and 136.50±27.39 ng×L⁻¹ respectively) (P < 0.01). AM from small cell lung cancer patients produced higher levels of IL-10 than squamous cell carcinoma and adenocarcinoma did (165.33±23.78 vs 127.74±26.19 ng×L⁻¹ and 120.30±29.66 ng×L⁻¹ respectively) (P < 0.01); Size of mass and performance status greatly affected the level of IL-10; IL-10 level of lung cancer patients with survival time more than 2 years was remarkably lower than that of patients with survival time less than 2 years (P < 0.01).
Higher level of IL-10 presents both in local and general body of lung cancer patients. IL-10 may play an important role in deterioration of lung cancer. Detection of IL-10 level may be helpful to evaluate cellular immunity and predict prognosis of lung cancer patients.
Zhongguo fei ai za zhi = Chinese journal of lung cancer 04/2004; 7(2):158-60.
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Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology 01/2003; 10(6):464-5.
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ABSTRACT: To investigate the effect of curcumin on the proliferation, cell cycle distribution and apoptosis of hepatocarcinoma cell line QGY.
The MTT method was used to assay the biologic activities of curcumin in different times and different doses. The cell cycle distribution was detected by flow cytometic analysis. The cell ultrastructure was observed by electronic microscopy.
Curcumin could inhibit effectively QGY in a dose- and time- dependent manner. IC(50) of curcumin to QGY was 49.50 micromol/L in 72 hours. The cell growth was arrested at S stage. Curcumin could lead to the degeneration, necrosis, and apoptosis.
Curcumin can interrupt the cell cycle and has a role in cytotoxicity, antiproliferation and inducing apoptosis of QGY.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology 01/2003; 10(6):449-51.
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Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology 05/2002; 10(2):160.
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ABSTRACT: To study the cell cycle block and apoptosis induced by hydroxycamptothecine (HCPT) in cultured rabbit lens epithelial cells (RLECs) and its affect on the gene expression of fas/fas-L in RLECs.
Flow cytometry was used to examine HCPT-induced cell cycle block in cultured RLECs. We observed cell morphological changes with light microscope and electron microscope. Apoptosis was detected by terminal deoxyribose transferase-mediated deoxy-uridine triphosphate nick end labeling (TUNEL) assay. The TUNEL assay was performed following the manufacturer's instruction of in situ cell death detection. Expression of Fas and Fas-L was estimated by immunohistochemical staining.
The flow cytometric analysis showed that the HCPT induced cell cycle block was at S phase. The ratio of nucleus and cytoplasm was increased. The electron microscopic examinations demonstrated that the HCPT initiated apoptosis with typical morphological features. The apoptosis of RLECs treated with HCPT was observed by TUNEL assay, and the number of the cells with apoptosis was increased with the increase of the HCPT concentration. The HCPT did not affect the gene expression of Fas/FasL.
HCPT not only induces mitotic block but also induces apoptosis of RLECs which is not mediated by the Fas/FasL signaling pathway.
[Zhonghua yan ke za zhi] Chinese journal of ophthalmology 04/2002; 38(3):161-4.
