Animals ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Endothelial Cells ; cytology ; drug effects ; physiology ; Female ; Humans ; Mice ; Mice, Inbred C57BL ; Neovascularization, Physiologic ; drug effects ; Vascular Endothelial Growth Factor A ; pharmacology

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Objective To compare effection of two kinds of bowel preparation for enteroscopy in aged patients. Methods 100 patients who would receive enteroseopy were divided into two groups. 50 cases in each group. Patients in experimental group received colon dialysis, patients in control group received oral medication. Observed the effectiveness of two kind of bowel preparations and adverse reaction. Results The effectiveness of colon dialysis was superior to oral medication(P <0. 05) ,there was no significant deviation the adverse reaction of two groups. Conclusions Colon dialysis is a suitable method in bowel preparation for enteroscopy in aged patients.

Objectives:To study the expression of GST-pi and MDR1 genes in operative specimens of ovarian cancer,and to analyze the possible clinical role of GST-pi and MDR1. Methods:Eighteen frozen specimens of ovarian carcinoma and ten specimens of normal ovarian tissues from patients were examined for the expression of GST-pi and MDR1 genes by means of RT-PCR, and quantitative analysis was performed using β-actin as internal contrast.Results: Positive expression rate of GST-pi and MDR1 in ovarian carcinoma were 61.1% and 33.3%,respectively,and in contrast, 20% and 10% in normal ovarian tissues respectively. The level of GST-pi gene expression in ovarian carcinoma was obviously higher than that in normal ovarian tissue (P<0.05)and MDR1 gene also had high level expression in ovarian carcinoma, but had no statistical significantance. Four patients with ovarian carcinoma had GST-pi and MDR1 coexpression. Expression levels of GST-pi mRNA were lower than that of protein. Conclusions: (1) GST-pi and MDR1 had higher level expression in ovarian carcinoma than in normal ovarian tissues. (2) GST-pi and MDR1 may have same regulating factors but different mechanisms of action. (3)Processing after transcription and/or regulation of translation level may exist in GST-pi expression.

In this work, polyelectrolyte microcapsules containing gold nanoparticles were prepared via layer by layer assembly. Gold nanoparticles and poly (allyamine hydrochloride) (PAH) were coated on the CaCO3 microparticles. And then EDTA was used to remove the CaCO3 core. Scanning electron microscopy (SEM) was used to characterize the surface of microcapsules. SEM images indicate that the microcapsules and the polyelectrolyte multilayer were deposited on the surface of CaCO3 microparticles. FITC-bovine serum albumin (FITC-BSA, 2 mg) was incorporated in the CaCO3 microparticles by co-precipitation. Fluorescence microscopy was used to observe the fluorescence intensity of microcapsules. The encapsulation efficiency was (34.31 +/- 2.44) %. The drug loading was (43.75 +/- 3.12) mg g(-1).
Calcium Carbonate ; chemistry ; Capsules ; Drug Carriers ; Drug Delivery Systems ; methods ; Electrolytes ; chemistry ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; chemistry ; Gold ; chemistry ; Microscopy, Electron, Scanning ; Microscopy, Fluorescence ; Nanoparticles ; Particle Size ; Serum Albumin, Bovine ; chemistry

OBJECTIVETo study the influence of multiple myeloma (MM) cells on normal endothelial cells in co-culture system in vitro.

METHODSA co-culture system of human MM cell line RPMI8226 with human umbilical vein endothelial cells (HUVECs) was established in vitro. Mono-cultured normal endothelial cells were used as control. Light microscopy and transmission electron microscopy were used to observe the morphology of the endothelial cells. The effects of HUVECs co-cultured with RPMI8226 on HUVECs angiogenesis were studied by modified transwell migration assay and net-like formation assay. The protein expression of brain derived neurotrophic factor (BDNF), TrkB, Endoglin, Tie-2, beta 3 integrin and vascular cell adhesion molecule-1 (VCAM-1) in HUVECs were determined by FACS and Western blot analysis, respectively.

RESULTSThe morphology of HUVECs co-cultured with RPMI8226 cells became a narrower apart of extended shape as they began to align themselves. The sizes of nucleus and nucleolus were enlarged with an increased ratio of nuclear to nucleoplasm. The endoplasm was lose and distorted and the number of surface microvilli decreased. The RPMI8226 cell stimulated the migration and net-like formation of HUVEC, the number of net-like structure and migration cell being increased by 112% and 136%, respectively, compared with that of mono-cultured HUVECs. The expressions of BDNF, TrkB, Endoglin, Tie-2, beta 3 integrin and VCAM-1 in the ECs co-cultured with RPMI8226 were all up-regulated in comparison with those in the controls.

CONCLUSIONThe MM cells promote formation of new vessels in co-cultured endothelial cells and the endothelial cells in MM are different from the normal ECs in character, and behavior.

Brain-Derived Neurotrophic Factor ; metabolism ; Cells, Cultured ; Coculture Techniques ; Endothelial Cells ; cytology ; Endothelium, Vascular ; cytology ; Humans ; Multiple Myeloma ; metabolism ; pathology ; Neovascularization, Physiologic ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo investigate the cytotoxic mechanism of cadmium (Cd) on cerebral cortical neurons.

METHODSThe primary cultures of rat cerebral cortical neurons were treated with different concentrations of cadmium acetate (0, 5, 10, and 20 micromol/L), and then the cell viability, apoptosis, ultrastructure, intracellular [Ca2+], and reactive oxygen species (ROS) levels, mitochondrial membrane potential (delta psi), activities of catalase (CAT) and superoxide dismutase (SOD) were measured.

RESULTSA progressive loss in cell viability and an increased number of apoptotic cells were observed. In addition, Cd-induced apoptotic morphological changes in cerebral cortical neurons were also demonstrated by Hoechst 33258 staining. Meanwhile, ultrastructural changes were distortion of mitochondrial cristae and an unusual arrangement. Simultaneously, elevation of intracellular [Ca2+]i and ROS levels, depletion of Delta Psi were revealed in a dose-dependent manner during the exposure. Moreover, CAT and SOD activities in the living cells increased significantly.

CONCLUSIONExposure of cortical neurons to different doses of Cd led to cellular death, mediated by an apoptotic mechanism, and the apoptotic death induced by oxidative stress may be a potential reason. And the disorder of intracellular homeostasis caused by oxidative stress and mitochondrial dysfunction may be a trigger for apoptosis in cortical neurons.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; Cerebral Cortex ; cytology ; drug effects ; metabolism ; In Vitro Techniques ; Neurons ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism

This study was aimed to further explore whether brain derived neurotrophic factor (BDNF) pathway is a potential therapeutic target in multiple myeloma (MM) and whether anti-BDNF monoclonal antibody can prevent the development of this disease. The in vivo antitumor effect of anti-BDNF monoclonal antibody (McAb) on a human myeloma xenograft animal model was evaluated. The model of xenograft tumors was established in the nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice by subcutaneous injection of human myeloma cell line RPMI8226. The antibodies were injected intraperitoneally at a dose of 20 microg/mouse at day 1, 2, 3 after inoculation or at a dose of 100 microg/mouse once a week after tumors were detected. The microvascular densities in tumors were analyzed by immunohistochemistry study. The effect of anti-BDNF McAb on the proliferation of RPMI8226 cells in vitro and on endothelial cells network formation in the co-culture system were determined by using a (3)H-thymidine incorporation assay and a Matrigel network formation assay, respectively. The results showed that multiple injections of anti-BDNF McAb reduced the tumor size, decreased the microvascular density and significantly prolonged tumor-free time and survival time. Moreover, the proliferation of RPMI8226 cells was inhibited in vitro by anti-BDNF McAb, but not by the control IgG. Anti-BDNF McAb also inhibited RPMI8226-induced network formation in endothelial cells in vitro. It is concluded that anti-BDNF monoclonal antibody can inhibit cell growth and angiogenesis in subcutaneous plasmacytoma.
Animals ; Antibodies, Monoclonal ; therapeutic use ; Brain-Derived Neurotrophic Factor ; immunology ; Cell Line, Tumor ; Humans ; Male ; Mice ; Mice, SCID ; Multiple Myeloma ; drug therapy ; metabolism ; pathology ; Neoplasms, Plasma Cell ; drug therapy ; Xenograft Model Antitumor Assays

OBJECTIVETo evaluate the in vivo antitumor effect of anti-brain derived neurotrophic factor (BDNF) monoclonal antibody (MoAb) on a human myeloma xenograft animal model.

METHODSThe xenograft tumor model was established in the nonobese diabetic/severe combined immunodeficiency (NOD/ SCID) mice by subcutaneous injection of human myeloma cell line RPMI 8226. The antibodies were injected intraperitoneally at a dose of 20 microg/mouse at day 1, day 2, day 3 after tumor cell inoculation or at a dose of 100 microg/mouse once a week after tumors were developed. The histologic and cytologic examination were performed to confirm the development of plasmacytomas. The microvascular densities (MVD) in tumors were analyzed by immunohistochemistry. The effect of anti-BDNF MoAb on the proliferation of RPMI 8226 cells in vitro and on endothelial cell network formation in the co-culture system were determined by 3H-thymidine incorporation assay and Matrigel network formation assay, respectively.

RESULTSThe xenograft NOD/SCID animal model had high capacity for growth of RPMI 8226 subcutaneous tumors and presented pathologic features of plasmacytomas. After subcutaneous injection of RPMI 8226 cells, all mice developed localized tumors in (20 +/- 2) d. On 20 microg anti-BDNF MoAb 3 consecutive treatment, the mean tumor-free time was extended to (30 +/- 6) d and survival was significantly prolonged compared with IgG-treated group [(57 +/- 7) d vs (48 +/- 4) d, P < 0.05]. When mice died naturally, the tumors size in anti-BDNF MoAb treated ones was also reduced compared with control group [(157.9 +/- 21.6) mm3 vs (405.5 +/- 35.2) mm3, P < 0.05]. When the antibody treatment (100 microg/mouse) underwent from 27 th to 60 day once a week after tumor inoculation, the local tumor growth was inhibited partially and necrosis and infiltration were observed in the tumors. The median MVD in the antibody-treated mice (100 microg/mouse) was 11 vessels/0.216 mm2. The IgG treated mice had no decrease in MVD of subcutaneous tumors compared with untreated mice. In vitro, anti-BDNF MoAb (1.5 microg/ml) significantly but partially inhibited HUVEC network formation induced by RPMI 8226 (68.2% reduction) and significantly inhibited RPMI 8226 proliferation, too. The IgG (1.5 microg/ml) treated mice had no significant effect on both of two assays.

CONCLUSIONSAnti-BDNF monoclonal antibody could inhibit growth and angiogenesis in subcutaneous myeloma tumors. BDNF is a potential therapeutic target in MM.

Animals ; Antibodies, Monoclonal ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Brain-Derived Neurotrophic Factor ; immunology ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Multiple Myeloma ; drug therapy ; metabolism ; Neovascularization, Pathologic ; drug therapy ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays