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

Our previous studies have demonstrated the effects of brain derived neurotrophic factor (BDNF) on promoting proliferation of multiple myeloma (MM) cells and inducing angiogenesis in MM in vitro. To further investigate whether the PI3K/Akt and MEK1/ERK pathway play a role in the BDNF-induced angiogenesis in vitro and to explore the further molecular mechanisms, two ways to establish human myeloma xenograft animal model were developed, their advantages and disadvantages were elucidated. The phosphorylation of AKT and ERK1/2 were detected in human umbilical vein endothelial cells (HUVECs) by Western blot. The angiogenic activity in vitro was evaluated by transwell migration assay and tubule formation assay. Cell proliferation was determined by crystal violet staining. Cell apoptosis was detected by FITC-Annexin-V/PI double staining and flow cytometry. The results showed that the BDNF activated the PI3K/Akt and MEK1/ERK pathway in the time-dependent manner. Ly294002 and PD98059 blocked the activation of Akt and ERK1/2 respond to BDNF. 100 ng/ml BDNF significantly increased HUVEC tube formation, migration and proliferation in vitro at a similar degree of 25 ng/ml VEGF. Furthermore, tube formation of HUVECs toward BDNF was significantly inhibited by 57% and 40% with 20 micromol/L Ly294002 and 20 micromol/L PD98059 treatment, respectively. At the same time, Ly294002 and PD98059 reduced the BDNF-induced migration of HUVECs by 74% and 36%, respectively. While BDNF-induced survival was only blocked by Ly294002 and BDNF-induced proliferation was only inhibited by PD98059. It is concluded that BDNF promotes angiogenesis of HUVECs in vitro. ERK and Akt are two crucial events in BDNF-mediated signal transduction leading to HUVECs angiogenesis by different mechanisms. Moreover, the latter is more important.
Angiogenesis Inducing Agents ; pharmacology ; Brain-Derived Neurotrophic Factor ; pharmacology ; Chromones ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; Flavonoids ; pharmacology ; Humans ; MAP Kinase Kinase 2 ; genetics ; metabolism ; Mitogen-Activated Protein Kinase 3 ; genetics ; metabolism ; Morpholines ; pharmacology ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; Umbilical Veins ; cytology

AIMTo observe the expression of Presenilin-1 (PS-1) and production of amyloid beta-protein (Abeta) in hippocampus of female senile rats and to investigate the effect of vitamin E(VE) on preventing Alzheimer's disease after menopause.

METHODSThe animal model was established using female senile rats. Experimental groups (n=8) were respectively given different doses of VE(5 mg/kg, 15 mg/kg, 60 mg/kg) per day. The expression of PS-1 in hippocampus was detected by immunohistochemistry, the level of Abeta in hippocampus was measured by Radioimmunoassay, and neuronal ultrastructure in hippocampal DG area was observed using transmission electron microscope.

RESULTSThe expression of PS-1 in rat hippocampus of senile control group was stronger than that of adult control group. PS-1 expressed weakly in three medication groups along with augmentation of dosage. The levels of Abeta were found to correlate statistically with the expression of PS-1. The content of Abeta in VE groups was significantly decreased compared to that in senile control group (P < 0.01). There were some changes in the neuronal ultrastructure of senile rats. Neurons were gradually recovered in VE groups.

CONCLUSIONVE may depress the production of Abeta by regulating the expression of PS-1, reducing neuronal injuries. VE may play a role in neuronal protection.

Aging ; Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Female ; Hippocampus ; drug effects ; metabolism ; Presenilin-1 ; metabolism ; Rats ; Rats, Wistar ; Vitamin E ; pharmacology

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

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