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.

Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. In order to investigate the role of sphingosine kinase-1 (SphK-1)/sphingosine 1-phosphate (S1P) signal pathway in the expression of CML cells, and to explore whether P210(bcr/abl) involved is activating SphK-1/S1P signal pathwey, the expressions of SphK-1 and S1P receptor mRNA in bcr/abl positive K562 cells and bcr/abl positive primary CML cells were detected by RT-PCR, the imatinib mesylate, the specific inhibitor of P210(bcr/abl) was employed to inhibit the P210(bcr/abl) tyrosine kinases of K562 cells and CML primary cells, and then the intracellular SphK-1 activity was assayed. The results indicated that after being cultured with 2.5 micromol/L imatinib mesylate for 0.5, 2, 6, 24 and 48 hours, the intensions of inhibiting SphK-1 activity were 0.007%, 38.9%, 34.6%, 28.1% and 76.1% resepectively. SphK-1 activity in CML cells also was reduced by 2.5 micromol/L imatinib mesylate (16.8% - 41.9% decrease). It is concluded that the CML cells express SphK-1 and different S1P receptor, and P210(bcr/abl) fusion protein in CML cells can activate SphK-1.
Benzamides ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Lysophospholipids ; genetics ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Signal Transduction ; genetics ; Sphingosine ; analogs & derivatives ; genetics ; metabolism

italic>Schisandra chinensis is a traditional Chinese medicine with the functions of reinforcing deficiency, strengthening, and inducing astringency, appliable to treat the chronic cough and deficiency in breath, palpitation, and insomnia, etc. A hybrid mass spectrometry scanning strategy (high-definition data-independent/data-dependent acquisition, HDDIDDA), enabling the ion mobility separation and alternating data-independent acquisition/data-dependent acquisition, was established, which, in combination with in-house library-driven automatic peak annotation workflows facilitated by the UNIFI software, was utilized to systematically characterize the multi-classes of chemical components from S. chinensis. The use of an HSS T3 column (100 mm × 2.1 mm, 1.8 μm), 0.1% formic acid in H₂O-acetonitrile as the mobile phase running at the flow rate of 0.3 mL·min^-1, and column temperature at 35 ℃, could enable good separation of the S. chinensis components within 42 min. HDDIDDA scan in both the positive and negative ion modes was employed for data acquisition. Based on the automatic peak annotation, reference standards comparison, MS² data interpretation, and literature analysis, we were able to identify or tentatively characterize 105 compounds in the S. chinensis decoction, involving 56 terpenoids, 42 lignans, five glycosides, one organic acid, and one flavonoid. HDDIDDA scanning can improve the coverage of data acquisition and improve the accuracy of identification, while CCS prediction analysis provides the possibility to distinguish isomers by the ion mobility technology. The results provide reference for the intelligent material basis research of TCM.

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

Acute-Phase Proteins ; metabolism ; Animals ; Female ; Hepatitis, Autoimmune ; immunology ; metabolism ; pathology ; Interleukin-17 ; metabolism ; Lipocalin-2 ; Lipocalins ; metabolism ; Liver ; pathology ; Mice ; Mice, Inbred BALB C ; Oncogene Proteins ; metabolism

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.

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

CD4+ T cells mainly interact with Sphingosine 1-phosphate (S1P) to regulate immune function through Sphingosine 1-phosphate receptor 1 (S1P1). This study was aimed to investigate the effects of recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization on S1P1 expression in CD4+ T cells of donor's peripheral blood. The CD4+T cells of peripheral blood were isolated by magnetic beads from 17 allo-hematopoietic stem cell transplantation (allo-HSCT) donors before and at fourth day of mobilization with rhG-CSF. The S1P1 expression was detected by real time quantitative PCR in the RNA extracted from CD4+ T cells collected before and after rhG-CSF mobilization. The results showed that the expression of S1P1 was found in CD4+ cells before and after rhG-CSF mobilization, but the expression level of SIP1 in CD4+ cells after rhG-CSF mobilization was significantly lower than that before rhG-CSF mobilization (p<0.01). It is concluded that the mobilization with rhG-CSF obviously down-regulates the expression of S1P1 in CD4+ T cells of donor's peripheral blood.
CD4-Positive T-Lymphocytes ; drug effects ; metabolism ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, Lysosphingolipid ; metabolism ; Recombinant Proteins

OBJECTIVETo elucidate association between the mutation of nuclear factor of activated T cells 1 (NFATC1) gene in IPT-NFAT region and simple congenital heart disease (CHD) in children.

METHODWe used polymerase chain reaction (PCR) and the sequencing reaction to detect the mutations on the patients and their parents and (or) siblings.

RESULTSPCR amplification of the exon 7 region showed that 2 bands are obtained in 58% of patients with CHD and in 74% of their healthy parents and (or) siblings. Sequencing of the 2 bands revealed that both are amplicons of the exon 7 region, and that the additional band harbors an additional 44 nucleotides segment in the intronic region. The homozygous form of this allele was only present in patients with ventricular septal defect (2/24), atrial septal defect (3/18) and bicuspid aortic valve (1/4) in which G to A transition at nucleotide 17 of the third 44 bps was found. Neither the unrelated non-CHD individuals nor the ones with other CHD showed positive presence for the homozygous form of this allele.

CONCLUSIONSThere is a differential amplification of a tandem repeat region in intron 7 of NFATC1 and homozygous form of this allele in patients with ventricular septal defect, atrial septal defect and bicuspid aortic valve. NFATC1 gene may be an a susceptibility marker for ventricular septal defect, atrial septal defect and bicuspid aortic valve.

Adolescent ; Adult ; Aged ; Base Sequence ; Child ; Child, Preschool ; Female ; Genetic Testing ; Heart Defects, Congenital ; genetics ; Humans ; Infant ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; NFATC Transcription Factors ; genetics ; Pedigree ; Young Adult