OBJECTIVETo analyze the relation between activation of NF-kappa B and chemotherapy induced apoptosis of leukemic cells and the effect of vincristine (VCR) on them.

METHODSElectrophoretic mobility shift assay (EMSA) was used to detect the activation of NF-kappa B and tunel DNA electrophoresis was adopted to observe the apoptosis induced by cytosine arabinoside (Ara-C) and etopside (Vp-16) in P388 leukemic cells.

RESULTSThe activation of NF-kappa B induced by Ara-C and Vp-16 was obviously correlated to apoptosis in P388 cells. VCR (0.1 micromol/L) could suppress activation of NF-kappa B by 52% and 63% and significantly increase the apoptosis by 89% and 123% as induced by Ara-C (100 micromol/L) and Vp-16 (100 micromol/L). The activity of NF-kappa B could be found in P388 cells before being exposed to chemotherapeutic agent.

CONCLUSIONChemotherapeutic agents can induce apoptosis and activation of NF-kappa B of P388 cells. The mechanism of VCR potentiating chemotherapeutics induction of leukemia cell apoptosis may be related to its suppression of the NF-kappa B activity in the P388 cells.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; In Situ Nick-End Labeling ; Leukemia P388 ; drug therapy ; pathology ; Mice ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Vincristine ; pharmacology

OBJECTIVETo apply conidia of Pyricularia Oryzae to the screening of antimitotic constituents from marine animal sea hare.

METHODTo extract and fractionate active portions from sea hare through detecting deformation of mycelia germinated from conidia of P. Oryzae P-2b, in comparison with the cytotoxic test results in vitro.

RESULTTwo active portions, of which IC50 against P388 and HL-60 was 23.4, 18.6 and 19.4, 12.5 micrograms.ml-1, respectively, were screened from this animal.

CONCLUSIONThis bioassay method was efficiently applied to the primary screening of antimitotic portions from marine animals for the first time. Being convenient, speedy and cheap, the screening model is suitable for the bioassay of active constituents from marine life.

Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Aplysia ; chemistry ; HL-60 Cells ; drug effects ; Humans ; Leukemia P388 ; pathology ; Materia Medica ; isolation & purification ; pharmacology ; Mice ; Mitosis ; drug effects ; Mitosporic Fungi ; physiology ; Tumor Cells, Cultured ; drug effects

AIMTo study the chemical constituents of Lycianthes biflora.

METHODSColumn chromatography was used to separate the chemical constituents. IR, MS, 1HNMR, 13CNMR and 2D-NMR technique were used to determine the structures of the isolated constituents.

RESULTSFive compounds were isolated from this plant. Their structures were identified to be bifloride A (1), N-trans-cinnamoyltyramine (2), liquiritigenin (3), N-trans-p-coumaroyloctopamine (4), 1-O-beta-D-glucopyranosyl-2-N-2'-hydroxypalmitoyl-sphinga-4- trans-8-trans-dienine (5).

CONCLUSIONCompounds 1 and 2 are new compounds, the others were isolated from this plant for the first time. Compound 2 showed inhibitory effects on P-388.

4-Butyrolactone ; analogs & derivatives ; chemistry ; isolation & purification ; Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; Cinnamates ; chemistry ; isolation & purification ; Leukemia P388 ; pathology ; Mice ; Molecular Structure ; Plants, Medicinal ; chemistry ; Solanaceae ; chemistry ; Tumor Cells, Cultured ; Tyramine ; analogs & derivatives ; chemistry ; isolation & purification

A new sesquiterpene hydroquinone (1) was isolated from a deep sea sediment derived fungus, Phialocephala sp.. Its structure and stereochemistry were established on the basis of spectroscopic data and optical rotation. This compound was tested for cytotoxicity against P388 (murine leukemia cell) and K562 (human leukemia cell) cell lines, and displayed strong cytotoxic effects with IC50 value of 0.16 and 0.05 micromol x L(-1), separately.
Animals ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Ascomycota ; chemistry ; Cell Line, Tumor ; Cell Survival ; drug effects ; Humans ; Hydroquinones ; chemistry ; isolation & purification ; pharmacology ; Inhibitory Concentration 50 ; K562 Cells ; Leukemia P388 ; pathology ; Mice ; Molecular Structure

This study was aimed to investigate a new method of avoiding graft-vs-host disease (GVHD) through selective elimination of alloreactive donor lymphocytes by using total body irradiation (TBI) and cyclophosphamide (Cy). Female (BALB/c x C57BL/6) F1 mice (H-2(d/b)) as recipients received (60)Co gamma-ray sublethal TBI of 4 Gy on day 0 followed by being inoculated with P388D1 leukemia cell line on day 1, injection of allogeneic splenocytes from C57BL/6 male mice (H-2(b)) was carried out for induction of graft-vs-leukemia (GVL) effect prior to stem cell transplantation (SCT), intraperitoneally injection of cyclophosphamide (Cy) (200 mg/kg) and TBI (9 Gy) was given on day 6. One day later, treated mice were rescued with bone marrow hematopoietic stem cells from (BALB/c x C57BL/6) F1 male mice (H-2(d/b)). The results showed that recipients had no occurrence of leukemia and GVHD through selective elimination of alloreactive donor lymphocytes by Cy and TBI, survived more than 210 days, the complete-donor chimerism occurred on day 21 after transplantation. The ratio of chimerism descended subsequently, but still displayed mixed-chimerism at 90 days. Control mice died of GVHD, leukemia or other death-related-transplantation within 20 to 36 days (P<0.01). It is concluded that to induce GVL effects by MHC mismatched splenocytes given before syngeneic bone marrow transplantation followed by selective elimination of alloreactive donor lymphocytes through TBI and Cy, graft-vs-host disease was thus avoided.
Animals ; Cyclophosphamide ; therapeutic use ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Tumor Effect ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Leukemia P388 ; therapy ; Lymphocyte Depletion ; Lymphocytes ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Whole-Body Irradiation

OBJECTIVETo study the influence of a discrete nano-hydroxyapatite crystal (nano-HAp) on lymphatic leukemia P388 behavior by in vivo techniques.

METHODSA nano-HAp was prepared by a neutralization reaction of 0.1 mol calcium hydroxide suspension and 0.06 mol phosphoric acid solutions at room temperature over pH7. The various doses of the nano-HAp only and the nano-HAp mixture with cyclophosphamide (CY) were injected into mice inoculated with solid tumor lymphatic leukemia P388 and dispersed into PRMI 1640 media harvested the leukemia P388 cells. Sixty P388 BALB/C mice were randomly grouped; 36 of them were used as nano-HAp treated groups and 24 mice as the control groups. The leukemia growth in the mice was examined morphologically, histopathologically and under a transmission electron microscope (TEM).

RESULTSThe nano-HAp was identified as a hydroxyapatite by an X-ray diffractometry (XRD) and a Fourier transform infrared spectroscopy (FTIR). The morphology and sizes were observed under a TEM. The tissue growth inhibition ratio (weight%) of solid lymphatic leukemia P388 bearing mice treated with nano-HAp at doses 35 mg/kg, 53 mg/kg and nano-HAp (53 mg/kg) combined with CY (35 mg/kg) in 3 consecutive days via intraperitineal injections were 14.95%, 32.67% and 60.45% respectively. Apoptosis of P388 cell cocultured with nano-HAp was confirmed by TEM.

CONCLUSIONSThe tissue growth restriction of solid tumor lymphatic leukemia P388 was greater after an injection of nano-HAp only or nano-HAp mixed with CY than that obtained after injection with physiological saline solution as a control (P < 0.01), and the tissue growth restriction of solid tumor after an injection of nano-HAp combined with CY was greater than that obtained after nano-HAp or CY injection only (P < 0.01).

Animals ; Biocompatible Materials ; pharmacology ; Calcium Hydroxide ; chemistry ; Cell Line, Tumor ; drug effects ; pathology ; Durapatite ; pharmacology ; Female ; Leukemia P388 ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Nanoparticles ; chemistry ; X-Ray Diffraction ; methods ; X-Rays

AIMTo synthesize eudistomin U and its 6-OCH3/Br derivatives and 5'-Br derivatives as antitumor agents.

METHODSUsing tryptamine and indole-3-aldehyde as starting materials, through condensation, Pictet-Spengler cyclization and dehydrogenation three steps, the alkaloids and its derivatives were prepared.

RESULTSThe structures of the compounds were determined by 1HNMR, MS and HRMS. Antitumor activity in vitro was tested.

CONCLUSIONEudistomin U and its derivatives were synthesized. The results showed that they all showed antitumor activities against mouse P388 strain.

Alkaloids ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Carbolines ; chemical synthesis ; chemistry ; pharmacology ; Cell Division ; drug effects ; Cell Line, Tumor ; Indoles ; Leukemia P388 ; pathology ; Mice ; Molecular Structure ; Tryptamines

To investigate the effect of bone marrow mesenchymal stem cells (BMMSC) on acute graft versus host disease (aGVHD) and graft versus leukemia (GVL) after allogeneic bone marrow transplantation (allo-BMT), both bone marrow cells and BMMSC obtained after three to four weeks of culture from donor mice were transplanted into the recipient mice injected with acute lymphocytic leukemia cells 5 days before, the control group was injected with bone marrow cells alone. The survial time after allo-BMT was recorded; the general manifestation and pathological changes of aGVHD in recipient mice were observed; the effects of BMMSC on the quatity of CD4(+) and CD8(+) T cell in vivo after allo-BMT were evaluated by flow cytometry; chimerism was detected by sex chromosome. The results showed BMMSC could increase obviously the survival time, and delay onset of aGVHD, BMMSC could decrease the amount of CD4(+) T cell and increase CD8(+) T cell in vivo. It is concluded that cotransplantation of bone marrow cells with BMMSC from the same donor mice has GVL effect. BMMSC can alleviate aGVHD and maintain GVL effect after allo-BMT.
Animals ; Bone Marrow Transplantation ; adverse effects ; methods ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Cell Line, Tumor ; Female ; Flow Cytometry ; Graft vs Host Disease ; etiology ; immunology ; prevention & control ; Graft vs Leukemia Effect ; immunology ; Leukemia P388 ; immunology ; pathology ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; adverse effects ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Transplantation, Homologous

This study was aimed to explore whether the GVHD in mice can be ameliorated and the GVL effect in mice can be reserved by transfusion of lymphocytes of donors fed with recipient splenocytes effect. Male (DBA-2) mice (H-2(d)) as donors were fed with BALB/c splenocytes, DBA-2 splenocytes, bovine serum albumin, or regular chow, every other day. Induction of tolerance was assessed by a mixed lymphocyte reaction (MLR). Female (BALB/c) mice (H-2(d)) as recipients received total body irradiation (TBI) of 6.0 Gy ((60)Cogamma-ray) followed by inoculation of 3 x 10(3) P388 mouse leukemia cells on the same day. Subsequently, tail vein injection of 2 x 10(7) splenocytes supplied by DBA-2 was undertaken. Control groups were fed identically without leukemia cell inoculation. The results showed that GVHD was significantly ameliorated and CD4(+)/CD8(+) ratio increased in recipient-mice transplanted with splenocytes of tolerated donors, compared with control animals. There was no significant difference in survival rate between different groups of recipients inoculated with leukemia cell. It is concluded that the peroral recipient-mouse splenocytes can ameliorate GVHD without hampering effect on GVL.
Adjuvants, Immunologic ; pharmacology ; Animals ; Cell Extracts ; immunology ; pharmacology ; Cell Transplantation ; Female ; Graft vs Host Disease ; immunology ; prevention & control ; Graft vs Leukemia Effect ; immunology ; Leukemia P388 ; therapy ; Lymphocytes ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred DBA ; Spleen ; cytology ; immunology ; Whole-Body Irradiation

OBJECTIVETo observe the pharmacodynamic and side effects of Wulong Kangai, a new drug of Chinese traditional herbal medicine, on 4 strains of mice transplantable tumors.

METHODMice transplantable tumors S180, H22, P388 and Lewis were used in the pharmacodynamic test on the granules of Wulong Kangai. The test on each tumor strain was repeated three times. In each test, 50 mice were used and divided into 5 groups. They were negative control group treated by physiological saline, cyclophosphamide control group and 3 test groups treated respectively with Wulong Kangai at deferent dosages of 10, 25, 40 g x kg(-1) x d(-1) in the treatment of Lewis and P388 and 15, 30, 50 g x kg(-1) x d(-1) in the treatment of S180 and H22.

RESULTThe tumor weight were inhibited at the rates of 90.1%, 30.8%, 49.8% and 52. 3% in the mice with tumors of Lewis, P388, S180, and H22 by high dosage of Wulong Kangai as compared with negative control group. The inhibitory rates in cyclophosphamide groups were 90.6%, 77.2%, 79.6% and 60.3% respectively. The mice body weights grew slower in high dose groups treated by Wulong Kangai granule.

CONCLUSIONWulong Kangai was effective in treating mice transplantable tumors of Lewis, P388, S180 and H22 with a dose-dependent manner. The Lewis was the most sensitive strain to the drug among the 4 kinds of tested tumors. Side effects appeared during 9-11 days of uninterrupted treatment with high dose Wulong Kangai.

Animals ; Antineoplastic Agents ; pharmacology ; toxicity ; Arthropods ; chemistry ; Carcinoma, Lewis Lung ; pathology ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; toxicity ; Female ; Leukemia P388 ; pathology ; Liver Neoplasms, Experimental ; pathology ; Male ; Materia Medica ; isolation & purification ; pharmacology ; toxicity ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; Plants, Medicinal ; chemistry ; Sarcoma 180 ; pathology