This study was performed to investigate the chemical constituents in the twigs and leaves of Harrisonia perforate. Six compounds were isolated from the 95% EtOH extract of the twigs and leaves of Harrisonia perforate by silica gel, ODS, Sephadex LH-20 column chromatographies and preparative HPLC. On the basis of chemical properties and spectra data, these compounds were identified as harriperfin E (1), kihadanin A (2), kihadanin B (3), 6α-acetoxyobacunol acetate (4), gardaubryone C (5), and β-sitosterol methyl ether (6), respectively. Compound 1 is a new chromone, and compounds 2-6 are isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Phytochemicals ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Simaroubaceae ; chemistry

BACKGROUND: Adipose-derived stem cells (ADSCs) represent one of the promising cell sources for myocardial regeneration due to their easy accessibility and efficacy in the improvement of cardiac function following myocardial infarction. However, previously reported studies on the underlying mechanism of ADSCs-mediated cardioprotective effect mainly focused on the ADSCs cultured at room air. OBJECTIVE: To test the paracrine actions and anti-apoptotic effect of ADSCs under hypoxic conditions. METHODS: After isolation and culture, neonatal rat myocardial cells were injured by hydrogen peroxide and co-cultured with rat ADSCs under normoxia and hypoxia (10% O2) conditions. Ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in the cell pellet and levels of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), and basic fibroblast growth factor (bFGF) were tested by ELISA. Expression of apoptotic proteins Bax and Bcl-2 were determined by western blot. RESULTS AND CONCLUSION: GSH/GSSG, VEGF, IGF-1, and bFGF were decreased in neonatal rat myocardial cells injured by hydrogen peroxide. ADSCs significantly attenuated hydrogen peroxide-induced myocardial apoptosis by increasing the ratio of GSH/GSSG and the secretion of VEGF, IGF-1 and bFGF. ADSCs also down-regulated Bax expression and up-regulated Bcl-2 expression. To conclude, hypoxic conditions can enhance the anti-apoptosis and cardioprotective effects of ADSCs through the paracrine mechanism.

The study aimed to explore the changes of the immunocyte quantities and cytotoxicity in bone marrow, and how many hematopoietic progenitor cells retained after the bone marrow cells were activated by different combinations of various cytokines. Bone marrow cells were divided into four groups and were cultured in vitro: (1) Control: no cytokines were added. (2) IL-2 group: bone marrow cells were activated by IL-2. (3) CD3-AK group: bone marrow cells were activated by IL-2 and CD3-McAb. (4) CIK group: IFN-gamma, IL-1, IL-2 and CD3-McAb were added. CFU-GM assay and CD3 phenotype detection were performed before and after activating culture in all groups. The changes of cell quantities during culture and cytotoxicity of cultured cells were tested. CD3 positive cells markedly increased in both CD3-AK and CIK groups. The cell numbers and cytotoxicity of CD3-AK and CIK groups were higher than those of control or IL-2 group obviously after culture (P < 0.05). CFU-GM were decreased in all groups after culture and there had no significant difference among four groups. The combination of IL-2 and CD3-McAb not only stimulates the proliferation of marrow immunocytes and increases their cytotoxicity but retains enough hematopoietic progenitor cells as well. This combination of cytokines can be used to purge autologous bone marrow in vitro.

To study immunophenotype and cytotoxicity of the immunocytes in bone marrow and peripheral blood after activation by combined cytokines, mononuclear cells (MNC) of bone marrow and peripheral blood were activated by IFN-gamma, IL-1, IL-2 and McAb-CD3 in vitro. The cell amount and morphology during culture were observed. Cytochemical staining and immunophenotype analysis were done before and after culture in two groups of MNC. Cytotoxicity was tested by MTT method. The results showed that the cell number of two groups increased obviously in culture (P < 0.05), while the peripheral blood mononuclear cells increased more markedly (P < 0.05). The cytochemical staining showed POX decrease, but PAS increase in two groups. The positive ratios of CD3(+), CD56(+) and CD38(+) cells in two groups increased obviously after culture (P < 0.05), but there was no significant difference between those two groups. CD3(+) CD56(+) cells increased obviously in peripheral blood mononuclear cells activated by cytokines (P < 0.05), but CD3(+) CD56(+) cells did not increase in bone marrow mononuclear cells. There was no significant difference between two groups' cytotoxicity. It was concluded that IFN-gamma, IL-1, IL-2 and McAb-C D3 increased cell number and cytotoxicity of both bone marrow and peripheral blood mononuclear cells that can be used in cell immunotherapy.
ADP-ribosyl Cyclase ; immunology ; ADP-ribosyl Cyclase 1 ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Bone Marrow Cells ; cytology ; drug effects ; immunology ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; Cell Count ; Cell Division ; drug effects ; Coculture Techniques ; Cytokines ; pharmacology ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Humans ; Immunophenotyping ; Interferon-gamma ; pharmacology ; Interleukin-1 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Membrane Glycoproteins ; Time Factors

OBJECTIVETo evaluate protective effects of Rheum tanguticum polysaccharides (RTP) on traumatic brain injury (TBI) in rats.

METHODThe polysaccharides (RTP) were extracted from Tanguficum Maxim. 120 rats were divided into 15 groups, with 8 rats in each group. RTP at 100, 200 and 400 mg x kg(-1) were administrated orally once a day for five days, and model of brain injury was made by dropping weight method.

RESULTRTP reduced water content and malondialdehyde (MDA) levels, and increased total SOD activity and Na+-K+ ATPase activity after injuried.

CONCLUSIONThe polysaccharides may be one of the effective comptents in Rheum tanguticum, showing significant neuroprotective effects.

Animals ; Brain Injuries ; enzymology ; metabolism ; pathology ; Cerebral Cortex ; enzymology ; ultrastructure ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism

This study was aimed to evaluate whether mesenchymal stem cells (MSCs) obtained from patients with myelodysplastic syndrome possess immunosuppressive effect. MSCs from bone marrow samples of MDS patients were isolated, cultured and expanded. MSCs were morphologically analyzed and their immunophenotype were determined by flow cytometry. Various amounts of MSCs were added into one-way mixed lymphocyte reaction. MSCs from MDS patients were tested for their ability to suppress in vitro proliferation of autologous and allogeneic peripheral blood lymphocytes (PBLs). The results showed that 3 x 10(3 - 1) x 10(5) MSCs from MDS patients could inhibit autologuous PBLs proliferation to (66.9 +/- 20.1)% - (30.2 +/- 5.9)% of maximal response, as well as inhibit allogeneic PBLs proliferation to (56.6 +/- 14.7)% - (20.5% +/- 9.7)% of maximal response, as compared with inhibitory ability of MSCs from healthy donors, there was no significant difference (P>0.05). It is concluded MSCs from patients with myelodysplastic syndrome also possess immunosuppressive activity.
Bone Marrow Cells ; immunology ; pathology ; Cell Proliferation ; Cells, Cultured ; Humans ; Immune Tolerance ; Lymphocyte Culture Test, Mixed ; Lymphocytes ; cytology ; Mesenchymal Stromal Cells ; immunology ; pathology ; Myelodysplastic Syndromes ; immunology ; pathology

The aim of this research was to understand the influence of rhG-CSF on the sphingosine kinase (SphK) activity of monocytes. The peripheral blood monocytes were collected from 6 peripheral blood progenitor cell donors on the fifth day of mobilization with rhG-CSF and from 5 blood donors' buffy coats. The mRNA expressions of monocyte G-CSF receptor and SphK were tested with RT-PCR. The changes of SphK activity of monocytes were assayed after being treated with rhG-CSF. The results showed that the two kinds monocytes collected from both blood donors and peripheral blood progenitor cell donors mobilized with rhG-CSF expressed mRNA of G-CSF receptor and SphK. The SphK activity of monocytes collected from blood donors was not changed significantly after being treated with rhG-CSF (P > 0.05). The SphK activity of monocytes collected from peripheral blood progenitor cell donors transiently increased by (39.6 - 87.2)% after being treated by means of rhG-CSF (P < 0.05) without obviously dose-dependent effect. It is concluded that the SphK activity of monocytes collected from peripheral blood progenitor cell donors can be activated by rhG-CSF.
Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Humans ; Monocytes ; cytology ; enzymology ; Phosphotransferases (Alcohol Group Acceptor) ; drug effects ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; biosynthesis ; genetics ; Recombinant Proteins

This study aimed to examine whether ophiopogonin D (OP-D) is capable of protecting cardiomyocytes against DOX-induced injury and the mechanisms involved. H9c2 cells were cultured. MTT assay was used to evaluate cell viability and toxicity. Mito-tracker as fluorescence probe was used to measure ROS content raised from mitochondria. The mRNA and protein expression of ATF6alpha, GRP78 and CHOP were analyzed using real-time PCR and Western blotting, respectively. The results showed that a significant endoplasmic reticulum stress (ERS) was induced upon exposure of H9c2 cells to DOX as indicated by the increase in the expression of ERS related proteins, which was paralleled with the accumulation of reactive oxygen species (ROS) and decrease in the viability of H9c2 cells. Whereas, DOX-induced ROS accumulation and up-regulation of ERS related proteins were partially abolished by pretreatment with OP-D. Consequently, a DOX-induced ERS was mitigated by application of OP-D. Similarly, DOX-induced decrease in cell viability was partially attenuated by either inhibiting CHOP or pretreatment with N-acetylcysteine (NAC), an antioxidant. Moreover, cardiac ultrastructural abnormalities seen in mouse receiving DOX injections were obviously ameliorated by pretreatment of OP-D. Taken together, the present study proved that OP-D protects cardiomyocytes against DOX-induced injury, at least in part, through reducing ROS accumulation and alleviating ERS.
Acetylcysteine ; Activating Transcription Factor 6 ; metabolism ; Animals ; Antioxidants ; Cell Line ; Cell Survival ; Doxorubicin ; adverse effects ; Endoplasmic Reticulum Stress ; drug effects ; Heat-Shock Proteins ; metabolism ; Mice ; Mitochondria ; metabolism ; Myocytes, Cardiac ; drug effects ; Rats ; Reactive Oxygen Species ; metabolism ; Saponins ; pharmacology ; Spirostans ; pharmacology ; Transcription Factor CHOP ; metabolism ; Up-Regulation

The study was to understand the impact on the proliferation and cytotoxicity of donor's T cells during mobilization with rhG-CSF. The peripheral blood mononuclear cells (PBMNC) were collected from 15 donors before mobilization and on fifth day of mobilization with rhG-CSF. After the PBMNC were activated with 500 ng/ml of CD3 monoclonal antibody and 500 microg/ml of rhIL-2 for 96 hours, the activated T cells were collected for testing proliferation, cytotoxicity, Fas expression, perforin and Fas ligand (FasL) mRNA expression, the IFN-gamma concentration in the culture medium of the activated T cells was determined by radioimmunoassay. The results showed that the proliferation activity of T lymphocytes and the cytotoxicity of T cells activated with CD3 monoclonal antibody and rhIL-2 were reduced markedly after mobilization with rhG-CSF (P < 0.05). The Fas molecule expression in the activated T cells was very high both before and after mobilization with rhG-CSF (P > 0.10). The activated T cells expressed perforin mRNA and didn't express FasL mRNA both before and after mobilization with rhG-CSF. The concentration of IFN-gamma in the culture medium of the activated T cells decreased significantly after mobilization with rhG-CSF (P < 0.01). It is concluded that activity of proliferation and cytotoxicity of donor's T cells is impaired after mobilization with rhG-CSF.
Adolescent ; Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fas Ligand Protein ; biosynthesis ; genetics ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; T-Lymphocytes ; cytology ; drug effects ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; fas Receptor ; biosynthesis ; genetics

The study was aimed to investigate the mobilization effect of medium dose of granulocyte colony stimulating factor (G-CSF) in allogeneic peripheral stem cell transplantation and changes of T lymphocyte subgroup in PBMNC before and after mobilization. G-CSF was administered at 600 microg/d (i.e. 300 microg i.v. twice a day) for successive 5 days to 31 matched sibling or unrelated donors for the mobilization. Stem cells were harvested on the fourth day. FACS was used to analyze the NC, MNC and T lymphocyte subgroups. The results showed that the number of NC, MNC, CD34(+) cells and CFU-GM in dose of 600microg/d significantly increased (P < 0.05), compared with 300 microg/d; the time for hematological reconstruction was significantly shortened (P < 0.05); the ratio of adverse effects was not obviously increased (P > 0.05) and the median percentage of CD3(+) lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization, while the ratio of CD4(+)/CD8(+) did not significantly changed. It is concluded that the administration of G-CSF 600 microg/d in allo-PBSCT has a good effect in the mobilization of PBSC with minor side effects, which can markedly promote hematopoietic reconstitution after transplantation. The relative amount of CD3(+) lymphocytes significantly decreased and the ratio of CD4(+)/CD8(+) remained unchanged, which may lead to alleviation of a GVHD after PBSCT.
Adolescent ; Adult ; Antigens, CD34 ; analysis ; Blood Donors ; Female ; Flow Cytometry ; Graft vs Host Disease ; prevention & control ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; methods ; Recombinant Proteins ; T-Lymphocyte Subsets ; cytology ; drug effects ; immunology ; Time Factors