1.Relationship Between Prolactin Cleavage and the Occurrence of Hypertension, Heart Failure in Post-menopausal Female Patients
Jiyang SONG ; Nan WANG ; Yan FANG ; Yunshan CAO
Chinese Circulation Journal 2015;(11):1058-1062
Objective: To investigate the relationship between prolactin (PRL) cleavage and the occurrence of hypertension, heart failure (HF) in post-menopausal female patients.
Methods: A total of 216 post-menopausal female subjects were enrolled in our study and they were divided into 3 groups: Hypertension group,n=80 patients with essential hypertension, Hypertension + HF group,n=76 and Control group,n=60 post-menopausal women form regular physical examination. The full length (23 kD PRL), 16 kD PRL fragment, lipid peroxide (LPO), total antioxidant status (TAS), left ventricular ejection fraction (LVEF), NT-proBNP, creatinine, uric acid, fasting blood glucose (FBG) and BMI were examined and compared among different groups.
Results:①There were no real differences in age, creatinine, uric acid, FBG and BMI among groups,P>0.05.②Compared with Control group, Hypertension group had increased LPO and decreased TAS; compared with Control group and Hypertension group, Hypertension + HF group had increased LPO and decreased TAS.③Compared with Control group, Hypertension + HF group showed lower level of 23KD PRL, higher level of 16KD PRL and the higher ratio of 16KD PRL/23KD PRL,P<0.05-0.01.④Pearson correlation analysis indicated that LPO was negatively related to 23KD PRL (r=-0.784,P<0.01), positively related to 16KD PRL (r=0.807, P<0.01); TAS was positively related to 23KD PRL (r=0.768, P<0.01), negatively related to 16KD PRL (r=-0.777P<0.01); 23KD PRL was positively related to LVEF (r=0.852, P<0.01), negatively related to NT-proBNP (r=-0.832P<0.01); 16KD PRL was negatively related to LVEF (r=-0.850,P<0.01), positively related to NT-proBNP (r=0.814,P<0.01).
Conclusion: PRL cleavage was highly related to the occurrence of hypertension and HF in post-menopausal female patients.
2.Arsenic trioxide induces apoptosis of gastric cancer cell AGS and influences STAT3 and VEGF expression
Fang ZHOU ; Yunshan WANG ; Yanfei JIA ; Anla HU ; Xiaoli MA ; Maoxiu ZHANG
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Objective: To investigate the apoptosis-inducing effect of arsenic trioxide (As2O3) on gastric carcinoma cell line AGS in vitro and to assess the influence of As2O3 on the expression of signal transducers and activators of transcription 3 (STAT3) and vascular endothelial growth factor (VEGF). Methods: AGS cells were treated with different concentrations of As2O3 (1, 5, and 10 ?mol/L) for 24,48, and 72 h. The cell proliferation was detected by MTT assay, cell apoptosis and cell cycle distribution were measured by flow cytometry and TUNEL, and the expression of STAT3 and VEGF was investigated by ELISA, immunohistochemistry and real-time PCR. Results: (1) As2O3 inhibited AGS cell proliferation in a time- and dose-dependent manner; (2) FCM results showed a typical sub-diploid peak before G0/ G1 phase and cell cycle analysis showed G2/M phase arrest; (3) TUNEL analysis revealed the DNA fragmentation; (4) During the As2O3-induced apoptosis of AGS cells, the expression of STAT3 and VEGF was down-regulated, especially when As2O3 was at 10 mol/L. Conclusion: As2O3 can inhibit the proliferation of AGS cells and induce AGS cell apoptosis, which might be related with cell cycle block and down-regulation of STAT3 and VEGF expression.
3.Effects of RATG on CD4+and CD8+ T cell eostimulatory molecule gene expression and productiun of immune-regulatory cytokines
Xiaoping WANG ; Zidong LIU ; Yusong FANG ; Geng WANG ; Liangming ZHU ; Yunshan ZHU ; He XU
Chinese Journal of Organ Transplantation 2008;29(9):526-530
Objective To investigate the immunological effects of thymoglobulin (RATG) on human CD4+and CD8+cells for costimulatory molecule gene expression and the production ofimmune-regulatory cytokines. Methods CD4+and CI8+T cells were isolated and purified fromnormal human peripheral blood mononuclear cells (PBMC) followed by incubation with RATG at37℃. Cells and culture supematants were collected at 24, 48, and 72 h after incubation, and analyzedby real-time quantitative polymerase chain reaction (RT-PCR) for CTLA-4, CD154, forkhead box P3(Foxp3), OX40, IFN-γ, IL-2, IL-10 and CD25 gene expression, and multiplex cytokine detectionassay for IFN-y, IL-2, IL-10, and IL-4 production. Untreated and rabbit isotype Ig-treated cells wereused as negative controls. Results RT-PCR demonstrated that RATG pre-treated CI+and CD8+cells upregulated the expression of CTLA-4, OX40, Foxp3, CD25, IFN-γ, IL-10 and IL-2 genes, anda dramatic increase of supernatant IFN-γ, IL-10, IL-2 and IL-4 was revealed 24 h after treatment asdetermined by multiplex cytokine detection assay when compared with negative controls. Theupre gulation of CTLA-4, Foxp3, OX40, IL-10 and CD25 was reduced, and a down-regulation ofCD154 and IL-2 gene expression was revealed 48 h after treatment. Cells, treated with RATG for 72h, demonstrated up-regulation of CTLA-4, Foxp3, OX40, IFN-y and CD25 gene expression, and theexpression of IL-2 and IL-10 genes was down-regulated. Additionally, supernatant IFN-γ, IL-2,IL-10 and IL-4 levels were decreased. Conclusion RATG stimulates CI4/CD8 T cells to up-regulatecostimulatory molecules and release immune regulation associated cytokines IF'N-γ, IL-2, IL-10in vitro. These results suggest that the unique effect of RATG on CD4+CD8+T cells may be animportant mechanism for its action in inducing immunoregulation, immunosuppression and transplanttolerance.
4.Detection of monocyte-derived costimulatory molecule expression and the role during discordant xenogeneic immune responses
Yusong FANG ; Zidong LIU ; Liangming ZHU ; Pu WANG ; Yunshan WANG ; He XU
Chinese Journal of Organ Transplantation 2008;29(10):589-593
Objective To explore the expression and the role of monocyte-derived costimulatory molecuels during xenogeneic immune responses. Methods Porcine endothelial cells (PEC) were isolated from aorta, and subcultures were performed. CD4+ cells and monocytes were purified from human peripheral blood mononuclear cells (PBMC). PBMC-PEC co-cultures were established, and the cells were collected followed by staining with florescent-labeled monoclonal antibodies and analyzing by FACS. In selected experiments, monoclonal antibodies specific for CD154, CD80 and CD86 were added into PBMC-PEC co-cultures, and the effects of co-stimulatory molecule blockade in inhibiting lymphocyte proliferation in response to PEC were determined by 3H-thymidine up-take. The proliferation of CD4+ cells induced by PEC-conditioned monocytes with or without co-stimulation blockade was evaluated. Results PBMC-PEC co-incubation demonstrated dramatic lymphocyte proliferation as determined by 3H-thymidine up-take. FACS found that resting monocytes expressed only CD86 but not CD40 and CD80. CD14+ monocytes from PEC-stimulated PBMC demonstrated up-regulation of CD80 and CD40 expression. The up-regulation of CD86 was revealed. PEC-activated monocytes induced CD4+ cell proliferation while resting monocytes did not and this proliferation was inhibited by anti-CD154, anti-CD80 or anti-CD86 antibodies. Conclusions CD14+ monocytes play an important role during xenogeneie immune responses in indirect antigen presentation and co-stimulation- The interaction between monocyte-derived co-stimulatory molecules and CD4+ cell-derived CD154 and CD28 delivers secondary signal and induces CD4+ proliferation, and the co-stimulation blockade inhibits xe-nogeneic cell-mediated immune responses.
5.A new generation of rabbit anti-human leukocyte polyclonal antibody in inhibiting xenogeneic cell-mediated immune pesponses
Liangming ZHU ; Yusong FANG ; Zidong LIU ; Xi WANG ; Xiuqing GUO ; Yunshan WANG ; Ke XU
Chinese Journal of Organ Transplantation 2009;30(2):103-106
Objective Delayed xenograft rejection (DXR) is a major barrier to the long-term xenograft survial.This study evaluated the interaction between human peripheral blood mononuelear cells (PBMC) and porcine endothelial cells (PEC),and the effects of new generation of rabbit antihuman leukocyte polyclonal antibody (newRALG) inhibiting xenogeneic cell-mediated immune responses.Methods newRALG was obtained from rabbits after immunization with activated lymphocytes and monoeytes.PEC were isolated from aorta,and human PBMC were isolated from peripheral blood.Co-cultures of PKH-26 labeled PEC with PBMC were established,newRALG,thymoglobulin,isotype Ig and scavenger receptor (SR) ligand poly G were added into the co-cultures.Cells were collected,then FACS analysis was carried out to detect the up-take of PEC membrane by monocytes and the expression of costimulatory molecules.Lymphocyte proliferative responses to PEC with or without antibody were evaluated by a xenogeneie mixed lymphocyte-endothelial cell reaction (xMLER).Results FACS analysis revealed that monocytes from PBMC-PEC co-cultures became positive for PKH-26 following their interaction with PKH-26 labeled PEC,indicating that they engulfed PEC membranes during activation.PKH-26 positive monocytes up-regulated the CD40 and CD80 expression.Furthermore,SR blockade with poly-G prevented PEC membrane up-take by monocytes,newRALG greatly reduced SR-mediated PEC membrane up-take.The effects of thymoglobulin in inhibiting PEC membrane uptake were limited.xMLER demonstrated strong lymphocyte proliferation in response to PEC,and lymphocyte proliferation was dramatically inhibited by newRALG but not isotype Ig at a dosmdependent manner.Conclusions Monocytes play an important role in xenogeneic immune responses.SR ligand poly G inhibits PEC membrane up-take.newRALG inhibits PEC membrane up-take by monocytes,suggesting that newRALG blocks SR.Additionally,newRALG inhibits lymphocyte proliferation in response to PEC.These results suggest that this new polyclonal preparation may thus impair the initiation of xeno-specific immune responses and prevent xenograft rejection.
6.Chemical constituents from the seed coat of Juglans regia.
Chuanshui LIU ; Zhigang TAI ; Siquan FENG ; Yunshan FANG ; Le CAI ; Zhongtao DING
China Journal of Chinese Materia Medica 2012;37(10):1417-1421
Fifteen compounds were isolated from the seed coat of Juglans regia by silica gel, MCI gel and Sephadex LH-20 gel column chromatography, as well as high preparative performance liquid chromatography. Their structures were identified as salidroside (1), (6S, 9S)-roseoside (2), (6S, 9R)-roseoside (3), blumenol C glucoside (4), byzantionoside B (5), 5-hydroxy-2-methoxy-1, 4-naphthoquinone (6), gallic acid (7), glycerol 1-(9Z-octadecenoate)-2-(9Z, 12Z-octadecadienoate)-3-(9Z, 12Z, 15Z-octadecatrienoate) (8), glycerol 1, 2, 3-tri-(9Z, 12Z-octadecadienoate) (9), glycerol 1, 2, 3-tri-(9Z, 12Z, 15Z-octadecatrienoate) (10), glycerol 1-hexadecanoate-2, 3-di-(9Z, 12Z-octadecadienoate) (11) on the basis of EI-MS, FAB-MS and NMR spectra. Moreover, 35 volatile compounds were identified by GC-MS.
Gas Chromatography-Mass Spectrometry
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Juglans
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chemistry
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Magnetic Resonance Spectroscopy
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Seeds
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chemistry
7.The mechanism research of Xiaoyan Lidan formula for the intervention of chronic intrahepatic cholestasis based on metabolomics combined with molecular docking analysis
Si-min CHEN ; Jin-hao HUANG ; De-qin WANG ; Yu-ying XIA ; Mei-qi WANG ; Run-feng SHI ; Fang-le LIU ; Chen-chen ZHU ; Chao-zhan LIN
Acta Pharmaceutica Sinica 2023;58(11):3408-3420
In this study, the mechanism of Xiaoyan Lidan formula (XYLDF) against 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-collidine (DDC)-induced chronic intrahepatic cholestasis (CIHC) in mice was investigated based on metabolomics, molecular docking and pharmacological methods. In the pharmacodynamics study, a dosage of 5 g·kg-1 (clinical equivalent) XYLDF was administered in DDC-induced mice, then the effect of XYLDF against CIHC was evaluated by measuring the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP) as well as total bilirubin (TBIL) in serum and observing liver histopathological changes. All experiments were approved by the Ethical Committee Experimental Animal Center of Guangzhou University of Chinese Medicine (ZYD-2021-001). The serum metabolites of mice in each group were detected and identified based on ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry, and the relevant biological pathways and molecular key targets were further enriched. Molecular docking technology was used to further evaluate the binding activity of the main active ingredients of XYLDF with potential targets. Subsequently, the
8.Immunosuppressive effects of fetal bone marrow derived mesenchymal stem cells on in vitro proliferation of adult peripheral lymphocyte and expression of immune-related factors.
Fang LI ; Junqiang LYU ; Yongjuan DUAN ; Yi SUN ; Dong LI ; Yunshan WANG ; Xiao HU ; Dongjie XIAO ;
Chinese Journal of Hematology 2014;35(10):891-896
OBJECTIVETo investigate the potential immunomodulatory properties of fetal bone marrow derived mesenchymal stem cells (FBM- MSCs).
METHODSMononuclear cells from the bone marrow of second trimester (14-22 wks) fetus were isolated and cultured for the derivation of MSCs. The derived FBM-MSC cells were characterized via morphology, immunophenotyping and the adipogenic and osteogenic differentiation assays. The immunomodulatory properties of FBM-MSC on lymphocytes were evaluated through the co- culture assay with PHA activated adult peripheral blood mononuclear cells (PBMCs).
RESULTSDerived FBM-MSCs were CD29⁺, CD44⁺, CD49e⁺, CD73⁺, CD90⁺, CD105⁺ and CD31⁻ , CD34⁻ , CD45⁻ , HLA-DR⁻ and can be differentiated into adipocytes and osteocytes. When co-cultured with PHA-activated PBMCs, FBM-MSCs inhibited the proliferation of lymphocytes up to 96% and down-regulated the secretion of inflammatory cytokines such as IFN-γ and TNF-α up to 90.9% and 58.4% respectively. When compared with FBM-MSCs cultured alone, the expression of MSCs derived immunomodulatory cytokines, such as IDO, TSG-6 and TGF-β, was up-regulated significantly in the co-culture system.
CONCLUSIONMSC derived from fetal bone marrow demonstrated immunosuppressive effects on adult PBMCs in vitro. MSC-derived cytokines like IDO, TSG-6 and TGF-β may be critical for FBM-MSCs mediated immunosuppressive function.
Adult ; Bone Marrow ; Bone Marrow Cells ; cytology ; immunology ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokines ; Hematopoietic Stem Cells ; Humans ; Immune Tolerance ; Immunophenotyping ; In Vitro Techniques ; Leukocytes, Mononuclear ; Lymphocytes ; Mesenchymal Stromal Cells ; cytology ; immunology ; Osteogenesis