Rat mesenchymal stem cells differentiate into neuron-like cells induced by berberine in vitro     

XIANG Ping  LI Hai-biao 《园艺学报》2004,20(1):51-53

AIM: To investigate whether berberine can induce rat mesenchymal stem cells (MSCs) to differentiate into neuron -like cells in vitro. METHODS: MSCs were separated from young rat femurs marrow and expanded in culture medium. MSCs were induced to differentiate by berberine. The morphological changes of MSCs were evaluated by light microscope.Neuron-spcific enolase (NSE), neurofilament (NF), glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. RESULTS: Induced by berberine for 8 hours,MSCs exhibited neurotype . The expression of NSE and NF in the neuron-like cells was positive, but the glial astrocyte marker GFAP didn't express. CONCLUSION: Berberine may induce adult rat MSCs to differentiate into neuron-like cells in vitro.  相似文献   

Analysis of blood biochemistry and arterial blood gas of systemic inflammatory response syndrome in the early time after trauma     

ZHOU Ze-qiang  YANG Feng  HAN Qing 《园艺学报》2004,20(9):1716-1718

AIM: To explore the variation of blood biochemistry and arterial blood gas of patients with systemic inflammatory response syndrome (SIRS) in the early time after trauma and improve the diagnosis and first aid. METHODS: Eighty-eight patients with trauma from August 2003 to February 2004 were divided into two groups by their AIS-ISS90 score. The data of temperature, pulse, respiratory rate, white blood cell counts, Hb, blood glucose and arterial blood gas (PaO₂, PaCO₂, HCO₃^-, AG) were collected and compared with each group by statistic methods. RESULTS: Of the 88 patients, 49 underwent SIRS, 12 in light trauma group (ISS≥16) and 37 in severe trauma group (ISS<16). Compared with light trauma group, the data of pulse, respiratory rate, white blood cell counts, blood glucose, AG and rate of SIRS of severe trauma group were higher, PaO₂ and HCO₃^- were lower and the cases of PaCO₂>45 mmHg or <35 mmHg were more (P<0.01). The data of temperature and Hb had not significant difference between two groups (P>0.05). 13 patients had MODS in severe trauma group and 2 died while none had MODS or died in light trauma group. CONCLUSION: Application of AIS-ISS90 and SIRS-related blood biochemistry and arterial blood gas is beneficial for the diagnosis and treatment of patients in the early time after trauma.  相似文献   

Inhibition of K562 cell growth by antisense drug targeting with VEGF mRNA in vitro     

FEI Jia  ZHANG Huan  HAN Zhong-chao 《园艺学报》2004,20(10):1769-1772

AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs.  相似文献   

Comparison of different conditions inducing embryonic stem cells     

QIAN Hai-yan  LI Geng-shan  XU Hong-xin  L&#  Jing-jun  JIANG Wen  WANG Jing 《园艺学报》2004,20(12):2198-2201

AIM: To evaluate the different conditions inducing mouse embryonic stem cells (ESC) in vitro to differentiate into cardiomyocytes. METHODS: BRL conditioned medium was used to promote the growth of ESC and maintain them in an undifferentiated state. During the inducing process, retinoic acid (RA), DMSO, activin-A and TGF-β₁ were used as inducing reagents, and made up six kinds of differentiating medium. Then a three-step method inducing ESC cultured in hanging drops, in suspension and in plating was used to induce the differentiation of ESC. RESULTS: ESC were induced in vitro to differentiate into cardiomyocytes. Of all groups, the highest differentiating rate was observed in the group induced by activin-A (20 μg/L) and TGF-β₁ (2 μg/L). CONCLUSION: The inducing conditions including activin-A (20 μg/L) and TGF-β₁ (2 μg/L) is very valuable in inducing ESC differentiation into cardiomyocytes.  相似文献   

Proteomic analysis of the effects of tumor necrosis factor-α on endothelial cells     

L&#  Jun-ping  WANG Shu-ren  MA Zeng-chun  WANG Sheng-qi 《园艺学报》2004,20(7):1121-1125

AIM: To investigate the affected proteins by tumor necrosis factor (TNF)-α in endothelial cells, and further explore the potential molecular mechanism of TNF-α on endothelial cells. METHODS: Nitric oxide (NO) production in the cultured human umbilical vein endothelial cells (HUVECs) was measured by a NO assay kit. Proteomic alterations were analyzed using two-dimensional electrophoresis, and peptide mass fingerprinting with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. RESULTS: NO production in HUVECs decreased significantly after TNF-α treatement. Proteomics analysis showed 21 protein spots were changed including 9 spots that were increased and 11 spots that were decreased after TNF-α stimulation, and 1 spot was only detected in TNF-α activated cell gels. CONCLUSIONS: Decreased the expression of ecNOS by TNF-α might result in decreased NO production. Up-regulated MAP/ERK kinase 3 expression might imply that TNF-α activates the expression of adhesion molecules. Cytoskeletal protein actin is also involved in TNF-α injuried HUVECs. Proteomic analysis can find some clues for identifying new potential target of TNF-α.  相似文献   

Human interleukin-1β induces A375-S2 human melanoma apoptosis through caspase pathway     

WANG Che  WANG Min-wei  TASHIRO Shin-ichi  ONODERA Satoshi  IKEJIMA Takashi 《园艺学报》2004,20(7):1138-1143

AIM: To study the molecular biological mechanism and signal transduction pathway of interleukin-1β (IL-1β)-induced apoptosis in A375-S2 melanoma cells. METHODS: Photomicrocropy showed typical apoptotic changes. The cytotoxic effect of IL-1β in vitro and influences of caspases in this effect were measured by MTT assay. The cytotoxicity of cells was assessed by LDH-based assay. Degradation of DNA was detected by agarose gel electrophoresis. RESULTS: The inhibitory effect of IL-1β on A375-S2 cell growth was in a dose and time-dependent manner, and cell death rate reached more than 90% at 72 h after treatment with 10^-9mol/L IL-1β. The inhibitors of caspase-family, -1, -3, -8, -9, and -10, partially blocked cell death at early stage. LDH assay showed that major IL-1β-induced cell death was apoptosis, and in a dose and time-dependent manner. Typical apoptotic DNA ladder was observed in agarose gel electrophoresis. CONCLUSION: IL-1β induced apoptosis in melanoma A375-S2 cells by activating caspase pathway.  相似文献   

The ex vivo expansion characteristic of endothelial progenitor cells     

LIU Chao  SUN Zong-quan  WU Yong-chao  CHEN Xin-zhong  FENG Jian-e 《园艺学报》2004,20(5):760-764

AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34⁺ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34⁺ and CD34^- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34⁺ and CD34^-,total MNC led to a significant increase in the expansion of CD34⁺ cells compared with CD34 enrichment (P＜0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P＞0.05). These differentiated EPCs were stained positive for CD34⁺, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34⁺ and AC133⁺cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34⁺ and CD34^- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs.  相似文献   

Bone marrow mesenchymal stem cells are induced to differentiate into cardiomyocytes by hepatocyte growth factor in vitro     

FANG Tian-ling  MIN Jun  DENG Xiao-geng  QIAN Shi-kun  CHU Zhong-hua  CHEN Ya-jin  SHAO Jing  WEI Jing  CHEN Ji-sheng 《园艺学报》2004,20(7):1167-1170

AIM: To explore a new method of hepatocyte growth factor (HGF) inducing bone marrow mesenchymal stem cells (MSC) to differentiate into cardiomyocytes. METHODS: Bone marrow MSC was cultured with DMEM media (10% fetal calf serum) 4-6 passages, and induced by HGF (10 μg/L) for 30 d. Automatical beating of the differentiated cells was observed daily with transverse microscopy, or under condition of 0.1% isoproterenol or cal-cium-deprived incubation. Specific cardiac myosin in the cells was indentified by immunochemistry. RESULTS: At 14-20 d of differentiation, bone marrow mesenchymal stem cells formed clones, in 10%-50% of which spontaneous beating cell-mass had come to continuously exist. Isoproterenol increased the beating rate and calcium-deprived media inhibited the beating. The cells were identified to be cardiomyocytes by expression of cardiac myosin heavy chain. CONCLUSION: HGF may induce bone marrow mesenchymal stem cells into cardiomyocytes with high efficiency, but the differentiating pathway of stem cells remains to be further studied.  相似文献   

Induction of tumor-specific cytotoxic T lymphocytes in vitro by dendritic cells pulsed with different glioma antigens     

ZHU Xin-mei  QIAO Jian  XIAO Bao-guo  LU Chuan-zhen 《园艺学报》2004,20(4):584-589

AIM: The goal of this study was to compare different methods for tumor antigen preparation, to observe the induction of tumor-specific cytotoxic T lymphocytes in rats by dendritic cells (DCs) pulsed with different tumor antigens. METHODS: The precursors of dendritic cells were isolated from bone marrow of rats, stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4). Then rat DCs were pulsed with C6 tumor cell antigens prepared with different methods: freeze-thaw, boiling or total protein extracted from ultrasonic crushed tumor cell. Subsequently primed DCs were cocultured with T lymphocytes isolated from spleen to induce CTL. Lymphocyte chemoattractant factor from DCs and cytokine IFN-γ release were determined by ELISA, the cytotoxicity of CTL was assayed by JAM test. RESULTS: DCs pulsed with boiled tumor cell in vitro induced an enhanced ability of T-cell proliferation and cytotoxic T lymphocyte activity.CONCLUSION: Our results demonstrated that DCs primed with boiled tumor cell may represent a method for inducing immune responses against the entire repertoire of tumor antigens of malignancies.  相似文献   

The mechanism of apoptosis induced by homoharringtonine in HL-60 cells     

CHEN Chun-yan  JIA Ji-hui  PAN Xiang-lin  ZHANG Qi  WANG Juan-dong  ZHOU Ya-bin 《园艺学报》2004,20(7):1183-1186

AIM: To study the signal transduction pathway of apoptosis initiation induced by homoharringtonine in HL-60 cells. METHODS: After establishing the model of apoptosis initiation induced by homoharringtonine in HL-60 cells, at the point of apoptosis initiation, molecular caspase-3, Bcl-2, Bax and Fas/FasL were measured with flow cytometry and transmission electron microscope. ERK2 and P38 expression in HL-60 cells were detected by using immunohistochemistry. RESULTS: The model of apoptosis initiation induced by homoharringtonine was established in HL-60 cells. At the point of apoptosis initiation, upregulation of caspase-3 and decrease in Bcl-2/Bax were observed. However, the expression of Fas/FasL did not significantly change. ERK2 expression decreased and P38 expression increased. CONCLUSIONS: Caspase-3, Bcl-2, Bax and mitogen activated protein kinase pathways were involved in signal transduction of apoptosis initiation induced by homoharringtonine in HL-60 cells.  相似文献