Adenocarcinoma ; pathology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Nucleosides ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; Pyridazines ; administration & dosage ; pharmacology ; Stomach Neoplasms ; pathology

Animals ; Fluorescence ; Humans ; Microscopy, Fluorescence ; Nanotechnology ; Neoplasms ; diagnosis ; Photons ; Quantum Dots ; Staining and Labeling ; instrumentation ; methods

OBJECTIVETo observe the effect of Huatuo Zaizao extractum (HTZZ) on focal cerebral ischemia/reperfusion (I/R) neurogenesis in rats induced by middle cerebral artery occlusion (MCAO) and its mechanism.

METHODTotally 55 healthy adult male Sprague-Dawley rats were divided into the sham operation group, the MCAO model group and HTZZ high, middle and low dose groups (5, 2.5, 1.25 g x kg(-1)), with 11 rats in each group, and orally administered with drugs. The focal cerebral ischemia model was established by performing a middle cerebral artery occlusion (MCAO, 90 min) followed by a seven-day reperfusion (once a day). The neurogenesis and expressions of extracellular signal-regulated kinase (ERK) and cAMP response element binding protein (CREB) were detected by the immunofluorescent staining. The enzyme linked immunosorbent assay (ELISA) was adopted to determine the vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF).

RESULTMCAO (90 min) followed by a seven-day reperfusion resulted in the significant increase in the number of penumbra cortex newborn neurons (BrdU(+) -NeuN(+)), which was accompanied by the growth of ERK and CREB phosphorylation and VEGF and BDNF levels. HTZZ could promote the generation of newborn neurons (BrdU(+)-NeuN(+)) and the ERK and CREB phosphorylation and increase VEGF and BDNF levels at the ischemic side.

CONCLUSIONHTZZ could promote the neurogenesis, which may be the interventional targets of effective traditional Chinese medicine Huatuo Zaizao extractum in promoting the self-repair function of the cerebral ischemic areas.

Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Neurogenesis ; drug effects ; Neurons ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo determine the content of imperatorin in the tetraploidy radix angelicae dahuricae, and compare it with the original diplontic varites.

METHODThe chromatographic method was carried out on Nova-pak (4.6 mm x 200 mm, 5 microns) column with acetonitrile-water solution as a mobile phase. The flow rate was 1 ml.min-1. the detection wavelength was at 248 nm, and the column temperature was 25 degrees C.

RESULTThe eontent of was imperatorin in the tetraploidy radix 0.460% and 0.225% imperatorin in the diplontic species.

CONCLUSIONThe content of the mainly active constituent in the tetraploid is double to what it is in the original diplontic species.

Angelica ; chemistry ; genetics ; Furocoumarins ; analysis ; Plant Roots ; chemistry ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Polyploidy

OBJECTIVETo observe the effect of small interfering RNA (siRNA)-induced MAPK p42 silencing on the survival of HeLa cells.

METHODSTwo siRNAs targeting at the MAPK p42 gene and one random siRNA were synthesized respectively by Silencer siRNA Construction Kit and transfected into HeLa cells by Lipofectamin 2000. The expression of p42(MAPK) in the transfected HeLa cells was analyzed by Western blotting and immunohistochemistry, and the morphology of cells were observed with electron microscope. TUNEL assay and Annexin V/PI staining were employed for detecting the cell apoptosis.

RESULTSThe expression of p42(MAPK) in the HeLa cells was remarkably suppressed after transfection with the two siRNAs, reduced by about 2.5 and 3.2 folds respectively in comparison with the negative control. Chromatin margination in the cell nuclei were observed in the transfected cells, and TUNEL assay and Annexin V/PI staining further confirmed the occurrence of cell apoptosis.

CONCLUSIONIn vitro MAPK p42 siRNA-1 and siRNA-2 transfection can specifically silence the gene expression and induce apoptosis of HeLa cells.

Apoptosis ; physiology ; Gene Silencing ; physiology ; HeLa Cells ; Humans ; Mitogen-Activated Protein Kinase 1 ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

Aim To investigate the effects of Ginkgo biloba extract(GBE)on vascular endothelial dysfunc-tion induced by chronic intermittent hypoxia(CIH)in rats and its related mechanisms.Methods The CIH rat model was established,and 50 and 100 mg·kg-1 GBE was administered by intragastric administration.The systolic blood pressure(SBP)of the tail artery was detected in each group.HE staining was used to detect the morphology of aorta tissue.DAF-FM DA staining and nitric reductase assay were used to detect NO levels.ELISA was used to detect serum ET-1,TNF-α and IL-6 levels.DHE staining was used to de-tect reactive oxygen species(ROS)levels of aortic tis-sue.Kits were used to detect the serum levels of MDA,SOD and GSH-Px.Western blot was used to detect the levels of VCAM-1,ICAM-1,nucleus Nrf2,HO-1 and NQO1 of aortic tissue.Results GBE sig-nificantly decreased the levels of SBP,ET-1,ROS,MDA,VCAM-1,ICAM-1,TNF-α and IL-6,and sig-nificantly increased the levels of NO,SOD,GSH-Px,nuclear Nrf2,HO-1 and NQO1 in CIH rats.GBE sig-nificantly improved the histomorphology of aorta in CIH rats.Conclusions GBE can improve vascular endo-thelial dysfunction and reduce blood pressure in CIH model rats.The mechanism may be related to the acti-vation of Nrf2/ARE pathway and the inhibition of oxi-dative stress and inflammation by GBE.

BACKGROUNDTo explore the effect of human osteopontin (hOPN) on the proliferation, transmigration and expression of matrix metallproteinase-2 (MMP-2) and matrix metallproteinase-9 (MMP-9) in osteosarcoma (OS) cells in vitro.

METHODSThe prokaryotic-expression vector of hOPN was produced. hOPN was then subcloned into E. coli BL21 (DE3) cells and purified with ProBond trade mark Columns. The proliferation, cell cycle and the expression of cyclin A in OS cells were investigated by using MTT assay, flow cytometry and Western blot respectively. The transmigration of OS cells was checked by using transwell cell culture chamber. The micro-pore-filter-membrane system was used to study the chemiotaxis of hOPN to OS cells. The levels of total protein were examined according to Coomassie Brilliant Blue manuals. The expression of MMP-2 and MMP-9 were evaluated by detecting the volume of degradation of gelatin on SDS-PAGE gel.

RESULTSThe prokaryotic-expression vector of hOPN and purified hOPN protein were achieved hOPN promoted OS cells proliferation in a dose-dependent manner, and stimulated cyclin A expression in OS cells to accelerate cell division cycle. hOPN facilitated the trans-membrane migration of OS cells. hOPN also enhanced the secretion of MMP-2 and MMP-9 in OS cells.

CONCLUSIONhOPN could stimulate cyclin A expression in OS cells. hOPN has chemiotaxis to OS cells and increases their transmigration. hOPN enhances the secretion of MMP-2 and MMP-9 in OS cells.

Bone Neoplasms ; pathology ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; secretion ; Matrix Metalloproteinase 9 ; secretion ; Osteopontin ; Osteosarcoma ; pathology ; Sialoglycoproteins ; pharmacology

Transmembrane protein p185 (the product of Her2/c-erbB-2 gene) is a member of the epidermal growth factor receptor (EGFR) family. Its overexpression was found in about 30% of breast cancer. It is essential to obtain soluble extracellular domain (ECD) of p185, especially disulfide bond rich domains, for identifying the epitopes of anti-p185 antibodies and researching the interrelationship between the antigen and antibody. The disulfide bond rich domain I-II and domain IV of p185 ECD were amplified from plasmid pBabe/erbB-2 by PCR respectively. These two fragments were inserted into pGEX/4T-1 vector, transfected into E. coli Origami B (DE3) pLysS and expressed inductively by low concentration of IPTG and low temperature overnight. After the pressure lysis of cells, the supernatants were analyzed by SDS-PAGE and the result demonstrated that this GST-fusion protein was expressed solubly in the amount of 10-15 mg/L. By the ELISA, Western blot and other immunological assays, the fusion proteins and their GST cut-off derivates both showed binding activities with several anti-p185 antibodies respectively. These results indicated that it was a feasible and effectual method to express disulfide bond rich domain I-II and domain IV of p185 ECD and this method may also be used to express other disulfide bond rich proteins.
Antibodies, Monoclonal ; immunology ; Disulfides ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Receptor, ErbB-2 ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Solubility ; Transfection

Objective To evaluate the value of transesophageal echocardiography (TEE) in transthoracic minimally invasive device closure of ventricular septal defect(VSD).Methods A total of 164 cases of VSD were recruited as candidates to receive transthoracic minimally invasive device closure between January 2007 and October 2010,including 138 perimembranous VSDs,3 muscular VSDs and 23 supracristal VSDs.Among these groups,85 male patients were included.Four-champer view,five-champer view,left ventricular long-axis view,short-axis view and right ventricular inflow view were detected to evaluate the availability of device closure.Results A total of 152 cases(92.7%)were successfully closed with a device.All the patients were followed up more than 3 months arranged with a standard protocol.No complete atrioventricular block (CAVB) or associated valvular complications were observed.Three of the five cases with traced residual shunt after device closure closed spontaneously.Conclusion TEE plays an important role in transthoracic minimally invasive device closure of ventricular septal defect,which has been proved by the good follow-up results without CAVB and associated valvular complications.

This study was purposed to investigate the dynamically monitoring minimal residual disease (MRD) by flow cytometry (FCM) in patients with acute leukemia (AL) after complete remission and its relation with prognosis. From October 2010 to May 2012, 58 cases of AL (including 45 cases of AML and 13 cases of ALL) were regularly monitored for MRD in bone marrow by FCM and their bone marrow morphology was observed by light microscopy at the same time which continued to relapse or to follow-up deadline in the Department of Hematology, the First Affiliated Hospital of Zhengzhou University. Through average follow-up for 9 months (3 - 21 months), the average MRD level of patients with CR was got. And the prognostic value of MRD level at different time points in AL patients after CR was analysed and summarized. MRD ≥ 1% was defined as positive, otherwise, as negative. The results showed that the maximum and minimum MRD levels of 45 AML patients were 9.57% and 0.01% respectively, the average was 0.67%; the maximum and minimum MRD levels of 13 cases of ALL patients were 7.9% and 0.0016% respectively, the average was 0.99%. Among 44 cases after induction therapy, the relapse rate of MRD(+) group was 53.3% (8/15), the relapse rate of MRD(-) group was 10.3% (3/29), and the relapse rate of MRD(+) group was higher than that of MRD(-) group (χ(2) = 7.58, P = 0.006). Among 58 cases after the first consolidatory therapy, the relapse rate of MRD(+) group was 62.5% (5/8), the relapse rate of MRD(-) group was 16.0% (8/50), and the relapse rate of MRD(+) group was higher than that of MRD(-) group (χ(2) = 6.11, P = 0.013). It is concluded that MRD detected by FCM has a large range (10(-6) - 10(-2)), which can not be used as a single indicator of complete remission. When MRD ≥ 1% after induction therapy and the first consolidatory therapy, the relapse rate significantly increases, MRD can be used as a sensitive indicator for prognosis.
Adolescent ; Adult ; Aged ; Female ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; pathology ; Prognosis ; Recurrence ; Remission Induction ; Young Adult