1.Necrotic cells induce and promote inflammatory responses in vitro.
Xuan LIU ; Yu-wei LUO ; Er-wei SUN
Journal of Southern Medical University 2009;29(4):659-662
OBJECTIVETo investigate the effect of necrotic cells on the secretion of inflammatory cytokines.
METHODSRAW264.7 macrophages and necrotic mouse thymocytes induced by heating were incubated for 18 h at a ratio of 5:1 in the absence or presence of lipopolysaccharide (LPS, 100 ng/ml). The supernatant of the cell culture was collected and the expression and secretion of the pro-inflammatory cytokines were measured using Bio-Plex suspension system.
RESULTSThe secretions of tumor necrosis factor-alpha (TNF-alpha) and interlukine-6 (IL-6) by macrophages co-cultured with the necrotic cells were significantly enhanced as compared with the control cells. The necrotic cells also significantly augmented the secretion of the pro-inflammatory cytokines induced by LPS.
CONCLUSIONNecrotic cells not only induces pro-inflammatory cytokine expression by themselves but also work synergistically with LPS to enhance the cytokine production, suggesting the important roles of necrotic cells to initiate and maintain the inflammatory responses.
Animals ; Cell Line ; Hot Temperature ; Inflammation ; etiology ; metabolism ; pathology ; Interleukin-6 ; secretion ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; drug effects ; metabolism ; Male ; Mice ; Necrosis ; complications ; Tumor Necrosis Factor-alpha ; secretion
2.Effects of apoptotic lymphocytes on proinflammatory cytokine secretion by hepatic sinusoidal endothelial cells.
Dong HE ; Yu-Wei LUO ; Li MA ; Er-Wei SUN
Journal of Southern Medical University 2009;29(6):1127-1129
OBJECTIVETo investigate the effects of apoptotic lymphocytes on the secretion of cytokines by hepatic sinusoidal endothelial cells (HSEC).
METHODSHuman HSEC cells were co-cultured for 16 h with allogenetic apoptotic lymphocytes induced by UVB irradiation. The supernatants were collected and the levels of interleukin-2, interferon-gamma, and tumor necrosis factor-alpha were detected by Luminex technique.
RESULTSAll the cytokines were down-regulated by about 50% in HSECs after co-culture with the apoptotic lymphocytes as compared with those in the control group (P<0.05).
CONCLUSIONSCo-culture with apoptotic lymphocytes can down-regulate the secretion of pro-inflammatory cytokines in HSECs, which may contribute to tolerogenic microenvironment in the liver.
Apoptosis ; physiology ; Cells, Cultured ; Coculture Techniques ; Cytokines ; secretion ; Down-Regulation ; Endothelial Cells ; cytology ; metabolism ; Humans ; Immune Tolerance ; Interferon-gamma ; secretion ; Interleukin-2 ; secretion ; Liver ; cytology ; Lymphocytes ; cytology ; Tumor Necrosis Factor-alpha ; secretion
3.In vivo tracing of transferred apoptotic cell labeled using CFSE: a flow cytometry-based assay method.
Yan WANG ; Yi GAO ; Er-wei SUN ; Jin-min XIE ; Hui-ying ZHANG ; Jian-bo CHEN
Journal of Southern Medical University 2006;26(5):599-602
OBJECTIVETo establish an assay method for detecting the migration of transferred apoptotic cells into the recipient using flow cytometry.
METHODSSpleen lymphocytes were isolated and labeled with an intracellular amine dye, carboxyfluorescein diacetate succinimidyl ester (CFSE), to allow discrimination. The labeled cells were induced with dexamethasone to undergo apoptosis and transferred into recipient mice via tail venous transfusion. Flow cytometry and histological examination of different tissues were performed at different time points. The stability of CFSE labeling for apoptotic cells was also tested.
RESULTSThe CFSE-labeled apoptotic cells were highly fluorescent with a positive labeling rate of (98.0+/-1.9)%. The stability of CFSE-labeling was testified, and the CFSE-labeled apoptotic cells entering different tissues at different time points were detected by flow cytometry and verified by histological examination.
CONCLUSIONFlow cytometry using CFSE labeling is reliable, sensitive, precise and convenient for apoptotic cell tracing in vivo and in vitro.
Adoptive Transfer ; methods ; Animals ; Apoptosis ; Dexamethasone ; pharmacology ; Female ; Flow Cytometry ; methods ; Fluoresceins ; chemistry ; pharmacokinetics ; Fluorescent Dyes ; chemistry ; pharmacokinetics ; Lymphocytes ; chemistry ; cytology ; drug effects ; Mice ; Mice, Inbred BALB C ; Reproducibility of Results ; Spleen ; cytology ; Succinimides ; chemistry ; pharmacokinetics
4.Does exposure to computers affect the routine parameters of semen quality?
Yue-Lian SUN ; Wei-Jin ZHOU ; Jun-Qing WU ; Er-Sheng GAO
Asian Journal of Andrology 2005;7(3):263-266
AIMTo assess whether exposure to computers harms the semen quality of healthy young men.
METHODSA total of 178 subjects were recruited from two maternity and children healthcare centers in Shanghai, 91 with a history of exposure to computers (i.e., exposure for 20 h or more per week in the last 2 years) and 87 persons to act as control (no or little exposure to computers). Data on the history of exposure to computers and other characteristics were obtained by means of a structured questionnaire interview. Semen samples were collected by masturbation in the place where the semen samples were analyzed.
RESULTSNo differences in the distribution of the semen parameters (semen volume, sperm density, percentage of progressive sperm, sperm viability and percentage of normal form sperm) were found between the exposed group and the control group. Exposure to computers was not found to be a risk factor for inferior semen quality after adjusting for potential confounders, including abstinence days, testicle size, occupation, history of exposure to toxic substances.
CONCLUSIONThe present study did not find that healthy men exposed to computers had inferior semen quality.
Adult ; Case-Control Studies ; China ; Computers ; Electromagnetic Fields ; adverse effects ; Humans ; Male ; Semen ; Surveys and Questionnaires
5.Effects of decitabine on proliferation and apoptosis of NB4 and K562 cells.
Xin-Ai HAN ; Hui-Lan ZENG ; Yan-Ping HAN ; Er-Wei SUN
Journal of Experimental Hematology 2013;21(2):356-360
This study was aimed to investigate the effects of decitabine (DAC) on proliferation and apoptosis of leukemia NB4 and K562 cells. The proliferation inhibition of DAC on NB4 and K562 cells was detected by Trypan blue staining. After treatment of DAC at different concentrations, the changes of cell cycle and CD11b expression was determined by flow cytometry. The cell morphological changes were observed by Wright's staining. The DNA ladder was used to detect cell apoptosis. The results indicated that DAC significantly inhibited the proliferation of NB4 and K562 cells in dose-and time-dependent manner. The median inhibitory concentration (IC50) of DAC-treated NB4 and K562 cells for 72 h was 0.113 µmol/L and 0.138 µmol/L, respectively. After treating these two cell lines with DAC at different concentration for 72 h, the cell ratio in G0/G1 phase significantly increased, while the cell ratio in S phase obviously decreased in 0.15 µmol/L DAC group (P < 0.05). The expression levels of myeloid differentiation antigen CD11b of both cell lines significantly increased in contrast to the control group (P < 0.05). The cell morphology detected by Wright's staining displayed partial differentiation and apoptosis after treating NB4 and K562 cells with DAC for 48 h. Typical apoptotic DNA ladder was observed in 0.15 µmol/L DAC group at 48 h. It is concluded that DAC can inhibit NB4 and K562 cell proliferation, induce cell differentiation and apoptosis, but more obviously for NB4 cells.
Apoptosis
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drug effects
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Azacitidine
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analogs & derivatives
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pharmacology
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Cell Differentiation
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drug effects
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Humans
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K562 Cells
6.The phagocytosis of apoptotic allogeneic lymphocytes by recipient liver antigen-presenting cells.
Yan WANG ; Yi GAO ; Er-wei SUN ; Zhong-hua CHEN
Chinese Journal of Hepatology 2006;14(5):370-374
OBJECTIVETo track the location of the transfused apoptotic allogeneic lymphocytes and asses the process of their accumulation and phagocytosis removal as consequences on allograft tolerance in recipient mice.
METHODSDonor spleen lymphocytes were labeled by CFSE and induced to apoptosis by dexamethasone incubation. After purification by anti-annexin V-conjugated magnetic beads isolation, apoptotic lymphocytes were transfused into recipient mice through the tail veins. Tissue samples from various organs were taken at various time points to analyze the fates of the apoptotic allogeneic lymphocytes and the phagocytosis of them by organ resident APCs.
RESULTSUsing fluorescent microscopy and flow cytometry, after the apoptotic cells were recognized and uptaken, the largest amount of labeled cells were accumulated in the livers and disappeared within not more than 12 hours. Recipient liver APCs were highly efficacious in phagocytosis of apoptotic allogeneic lymphocytes; the removal was completed within 15 minutes after incubation. LSEC, KC and LDC all phagocytosized the apoptotic allogeneic lymphocytes but with significantly different rates. Considering the numbers of those cells in a normal liver, it could be calculated that LSEC and KC had greater effects in this activity.
CONCLUSIONSThe liver deserves foremost attention for study of the mechanism of allografts tolerance induced by pre-transfusion of apoptotic donor spleen lymphocytes. LSEC and KC are the main functional APCs to the alloantigens.
Animals ; Antigen-Presenting Cells ; cytology ; immunology ; Apoptosis ; physiology ; Liver ; cytology ; immunology ; Lymphocyte Transfusion ; Lymphocytes ; cytology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Phagocytosis ; physiology ; Spleen ; cytology ; Transplantation, Homologous
7.Protecting effects of p44/42 MAPK signal transduction pathway on hepatocytes in ischemic preconditioning.
Yi GAO ; Yu WANG ; Ming-xin PAN ; Er-wei SUN ; Yu-qiang SHAN
Chinese Journal of Hepatology 2003;11(3):166-169
OBJECTIVETo investigate the significance of PKC and p44/42 mitogen-activated protein kinase (MAPK) signal transduction in ischemic preconditioning (IP).
METHODSThrough liver cell IP models, PKC inhibitor and MEK inhibitor were utilized to analyze the phosphorylation level of p44/42 MAPK and cell viability was also observed. Rat liver IP models were established which were treated with various drugs. Then the phosphorylation level of p44/42 MAPK in vivo and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) concentrations were detected. And cellular structures were observed under light microscopy.
RESULTSSimilar results were obtained in vivo and in vitro IP models. Compared with the ischemia reperfusion (IR) group in vivo, the phosphorylation level of p44/42 MAPK was obviously increased in IP treated rats (q = 27.217, P < 0.01), and the cellular structure injured slightly. The concentrations of serum ALT and AST in IP group were significantly lower than those in IR group (281.0 U/L +/-35.6 U/L vs 762.8 U/L +/-130.5 U/L and 407.7 U/L +/-73.7 U/L vs 820.9 U/L +/-111.3 U/L, P < 0.01). However, opposite changes were found in PKC and MEK inhibited groups, when compared to IP group. The phosphorylation level of p44/42 MAPK was obviously decreased, the liver tissues injured evidently, and the concentrations of serum ALT and AST (645.61 U/L +/-90.4 U/L, 678.6 U/L +/-136.5U/L and 466.2 U/L +/-82.8 U/L, 732.9 U/L +/-91.1 U/L, respectively) were significantly greater than those in IP group.
CONCLUSIONThese results suggest that p44/42 MAPK pathway plays a vital role in the protection of hepatocytes in ischemic preconditioning.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Hepatocytes ; cytology ; physiology ; Humans ; Ischemic Preconditioning ; Liver ; blood supply ; cytology ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction
9.Phage display screening of therapeutic peptide for cancer targeting and therapy.
Protein & Cell 2019;10(11):787-807
Recently, phage display technology has been announced as the recipient of Nobel Prize in Chemistry 2018. Phage display technique allows high affinity target-binding peptides to be selected from a complex mixture pool of billions of displayed peptides on phage in a combinatorial library and could be further enriched through the biopanning process; proving to be a powerful technique in the screening of peptide with high affinity and selectivity. In this review, we will first discuss the modifications in phage display techniques used to isolate various cancer-specific ligands by in situ, in vitro, in vivo, and ex vivo screening methods. We will then discuss prominent examples of solid tumor targeting-peptides; namely peptide targeting tumor vasculature, tumor microenvironment (TME) and over-expressed receptors on cancer cells identified through phage display screening. We will also discuss the current challenges and future outlook for targeting peptide-based therapeutics in the clinics.
10.Effect of FK506 on cytokine secretion in whole blood.
He YI ; Li MIN ; Zhan-guo LIU ; Yu-wei LUO ; Ming WANG ; Jie ZHANG ; Er-wei SUN
Journal of Southern Medical University 2010;30(2):249-251
OBJECTIVETo investigate the effect of FK506 on cytokine secretions in whole blood from healthy individuals.
METHODSBlood samples collected from healthy volunteers were co-cultured with different concentrations of FK506 and stimulated with PMA and IONO. The concentrations of 8 cytokines including IL-2, IL-6, IL-12, IL-17, IFN-gamma, TNF-alpha, GM-CSF and G-CSF were detected by Bio-Plex suspension system.
RESULTSCompared with the control group, high-concentration FK506 (20 ng/ml) significantly inhibited the secretions of IL-2, IL-6, IL-12, IL-17, IFN-gamma, TNF-alpha, GM-CSF and G-CSF. At a moderate concentration (5 ng/ml), FK506 inhibited the secretion of GM-CSF significantly.
CONCLUSIONFK506 effectively inhibits the secretion of proinflammatory cytokines including IL-6, IFN-gamma and TNF-alpha and also the secretion of IL-2, IL-12, IL-17, GM-CSF and G-CSF. FK506 might play the role of immunosuppression by inhibiting the production of these cytokines by the immune cells. Monitoring the levels of these cytokines might be a potential method for evaluating the adequacy of FK506 doses administered.
Adult ; Cytokines ; blood ; secretion ; Down-Regulation ; drug effects ; Female ; Humans ; Immunosuppressive Agents ; pharmacology ; Interferon-gamma ; blood ; secretion ; Interleukin-6 ; blood ; secretion ; Male ; Tacrolimus ; pharmacology ; Tumor Necrosis Factor-alpha ; blood ; secretion ; Young Adult