Objective To investigate the effects of pulsed electromagnetic fields(PEMFs)on proliferation and differentiation of endothelial progenitor cells(EPCs).Methods EPCs were isolated from rat bone marrow by density gradient centrifugation.EPCs were exposed to PEMFs from the 5th day to the end of culture.MTT was used to measure the proliferation of EPCs.The expression ofⅧ-related antigen and NOS_3 was evaluated by flow cytometry. Results Compared with the control,the proliferating ability of EPCs exposed to PEMFs was stronger;the number ofⅧ-related antigen and NOS_3 positive cells increased significantly in EPCs exposed in PEMFs.Conclusion PEMFs promotes the proliferation and differentiation of rat bone marrow EPCs.

OBJECTIVETo observe the improvement effects of puerarin on glycated brain damages in rat model induced by D-galactose.

METHODThe model rats of protein glycation were induced by intraperitoneal administration of D-galactose (150 mg x kg(-1) x d(-1)) for 8 weeks, and all rats were treated with puerarin (high dose 300 mg x kg(-1), middle dose 150 mg x kg(-1), low dose 75 mg x kg(-1)) for 6 weeks. The activity of aldose reductase in red blood cells, the amount of glycated products (fructosamine in serum, glycohaemoglobin, advanced glycation end-products) and AGEs in brain tissue, calcium ion in brain cells were measured. Moreover, mitochondria in brain hippocampus cells were observed under electronic microscope.

RESULTHigh dose and middle dose of puerarin can decrease the activity of aldose reductase in red blood cells (P < 0.01), and inhibit the formation of glycation products significantly in model rats induced by D-galactose (P < 0.01). Also, puerarin can decrease the content of AGEs in brain and the level of calcium ions in brain cells (P < 0.05, P < 0.01), and decrease lesions degree in mitochondria in brain hippocampus cells.

CONCLUSIONPuerarin can produce the protective effects on glycated brain damages through inhibiting the glycation reaction in rats induced by D-galactose.

Aldehyde Reductase ; metabolism ; Animals ; Brain ; metabolism ; pathology ; Calcium ; metabolism ; Erythrocytes ; enzymology ; Female ; Fructosamine ; blood ; Galactose ; antagonists & inhibitors ; Glycated Hemoglobin A ; metabolism ; Glycation End Products, Advanced ; metabolism ; Hippocampus ; ultrastructure ; Isoflavones ; isolation & purification ; pharmacology ; Male ; Mitochondria ; ultrastructure ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To study the status and influencing factors on drug therapy compliance for the spinal tuberculosis patients after leaving hospital, so as to advance the corresponding nursing intervention strategies and to consolidate the cure effect. Methods 168 spinal tuberculosis patients in community were investigated by the basic information schedule table, compliance with regular treatment schedule table, the degree of disease cognition questionnaire, and the social support questionnaire. All questionnaires were collected and statistically treated. Results The scores of drug therapy compliance for the spinal tuberculosis pateints after leaving hospital were in a lower level, and the rate of compliance with regular treatment for spinal tuberculosis patients was 46.8%. The drug therapy compliance was affected by the years of age, the degree of disease cognition, the adverse reaction of medicine, the economy condition, the degree of civilization and the degree of social support (P <0.05). The drug therapy compliance was positively correlated with the degree of disease cognition, the economy condition, the degree of civilization and the degree of social support. The drug therapy compliance was negatively correlated with the years of age and the adverse reaction of medicine. Conclusions The druy therapy compliance for the spinal tuberculosis patients out of hospital is generally in a lower level Nursing stuffs should adopt active nursing interventions to raise the drug therapy compliance for spinal tuberculosis patients based on the influencing factors.

OBJECTIVETo elucidate the interference effect of Epigallocatechin-3-Gallate (EGCG) on targets of Activator Protein-1 (AP-1) signal transduction pathway activated by EB virus encoded latent membrane protein 1 in nasopharyngeal carcinoma (NPC) cell lines.

METHODSSurvival rate of cells was determined by MTT assay. AP-1 and CyclinD1 activation were analyzed by promoter luciferase reporter system. Nuclear translocation of JNK was analyzed by indirect immunofluorescence. Protein expression and phosphorylation were observed by Western blot.

RESULTSEGCG inhibited the survival of CNE1 and CNE-LMP1 cells and the activity of AP-1 caused by LMP1 in CNE-LMP1 cells. EGCG also inhibited the nuclear translocation of JNK and the phosphorylation of c-Jun. It also inhibited cyclinD1 promoter activity and cyclinD1 expression.

CONCLUSIONEGCG inhibits AP-1, JNK, c-Jun and cyclinD1 which are key targets on AP-1 signal transduction pathway. The results may explain the molecular mechanism of action of EGCG against nasopharyngeal carcinoma.

Carcinoma, Squamous Cell ; metabolism ; pathology ; virology ; Catechin ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Herpesvirus 4, Human ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; MAP Kinase Kinase 4 ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; virology ; Phosphorylation ; drug effects ; Protein Transport ; drug effects ; Proto-Oncogene Proteins c-jun ; metabolism ; Signal Transduction ; drug effects ; Transcription Factor AP-1 ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism ; physiology

The study was purposed to investigate the effect of arsenic trioxide (As(2)O(3))- induced p16 gene demethylation by a sensitive and specific PCR-based method (nested-methylation specific PCR, n-MSP) and DNA sequencing for rapid analysis of the promoter demethylation status, and to explore the possible mechanism of the p16 gene demethylation in human multiple myeloma U266 cells induced by As(2)O(3). The methylation status of the p16 gene in U266 cell line before and after treatment with As(2)O(3) was detected by the nested-methylation specific PCR and DNA sequencing, the mRNA of p16, DNA methyltransferase (DNMT 1, DNMT3A and 3B) gene were determined by RT-PCR, and the induced growth inhibition of U266 cell was assayed by growth curve, MTT and CFU; the DNA content of U266 cells was analyzed by flow cytometry after being exposed to As(2)O(3). The results showed that (1) all cytosines in CpG dinucleotides in untreated U266 cell not were changed, while all cytosines in treated U266 cells with As(2)O(3) had been converted to thymidine. (2) p16 gene was not expressed in U266 cell line after methylation. As compared with the beta-actin, the expression of U266 cell p16 gene mRNA was increased to (0.22 +/- 0.10), (0.59 +/- 0.11), (0.68 +/- 0.09) after exposed to 0.5 micromol/L, 1.0 micromol/L and 2.0 micromol/L As(2)O(3) for 72 hours respectively. (3) As(2)O(3) could significantly down-regulate DNA methyltransferase 1 (DNMT 1), DNMT3A and DNMT3B gene at mRNA level in a dose-dependent manner. (4) U266 cells line grew slowly and arrested at G(0) - G(1) phase after treatment with three different concentrations of As(2)O(3). It is concluded that As(2)O(3) can activate and up-regulate the expression of p16 gene which inhibits the proliferation of U266 cell through inducing the G(0) - G(1) arrest by demethylation or/and by inhibiting DNMT 1, DNMT3A and 3B gene.
Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Base Sequence ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; biosynthesis ; genetics ; DNA Methylation ; drug effects ; Genes, p16 ; Humans ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Oxides ; pharmacology ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transcription, Genetic ; drug effects

OBJECTIVETo investigate the prevalence of JAK2V617F and MPLW515L/K mutation in patients with slightly elevated platelets (BPC) or hemoglobin (Hb) not meeting the criteria of polycythemia vera (PV) or essential thrombocythemia (ET).

METHODSGenomic DNA from bone marrow or blood mononuclear cells was screened with allele specific polymerase chain reaction (AS-PCR) for JAK2V617F and MPLW515L/K mutation. The history of thrombosis was assessed retrospectively by patients files.

RESULTSOf 30 patients, 14 (46.7%) were positive for the JAK2V617F mutation, none of them had the MPLW515L/ K. Five of these 14 patients had a history of thrombosis. Follow-up results were available in 22 patients. Among them, 12 patients with JAK2V617F mutation turned out to be MPD in 6-24 months; only 2 out of 10 patients without this mutation evolved to MPD.

CONCLUSIONJAK2V617F mutation could be one of the diagnosis criteria of early MPD. No MPLW515L/K expression was found in early MPD.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Early Diagnosis ; Female ; Follow-Up Studies ; Humans ; Janus Kinase 2 ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; diagnosis ; genetics ; metabolism ; Receptors, Thrombopoietin ; genetics ; metabolism ; Young Adult

This study was aimed to investigate the reversing effect of arsenic trioxide (As2O3) on methylation status and the regulatory effect on transcription of malignant lymphoma cell line CA46 p16 gene as well as their possibe mechanisms. The hypermethylated malignant lymphoma cell line CA46 was used as a subject of experiment for studying relation of gene methylation with expression. The effect of As2O3 on the proliferation and viability of CA46 was detected by SRB method, the change of p16 methylation status after exposure to As2O3 was determined by nMSP, the expressions of p16, DNMT1, DNMT3A, DNMT3B mRNA were assayed by RT-PCR, the influence of As2O3 on CA46 cell cycle was analyzed by flow cytometry using analytical method for DNA ploidy. The results showed that the methylation level of p16 gene was obviously reduced after treatment with As2O3 for 72 hours and the hypermethylation of p16 gene was successfully reversed; the expression of p16 gene in untreated (control) group was low while it was enhanced in treated groups; the gray scale ratios of p16 gene to beta-actin in groups treated with As2O3 of concentration 0.5, 1.0 and 2.0 micromol/L were 0.33+/-0.10, 0.57+/-0.11 and 0.67+/-0.09 respectively, exhibiting a significant difference in comparison with 0.73+/-0.13 of positive control (p<0.01); as compared with the untreated group, the expression of DNMT3A and DNMT3B in treated groups was obviously down-regulated in a concentration-dependent manner, while expression of DNMT1 was nearly unchanged; as compared with control, all the 3 different concentrations of As2O3 could inhibit the proliferation of CA46 cells and increase the cell number in G0/G1 phase. It is concluded that the As2O3 may up-regulate the expression of p16 gene, recover the activity of p16 gene, thereby promote the regulatory function on cell cycle resul-ting in arrest of cells in G0/G1 phase and inhibit growth of tumor cells through depressing the expression of DNMT3A and DNMT3B and/or directly reversing the methylation status of p16 gene.
Arsenicals ; pharmacology ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; drug effects ; Genes, p16 ; Humans ; Lymphoma ; genetics ; Oxides ; pharmacology ; Transcriptional Activation

To decrease the hemolysis side effect of puerarin, the basic formula and preparation of puerarin submicron emulsion were optimized and the physicochemical properties were evaluated. Puerarin submicron emulsions were prepared by phase inversion-ultrasound combining with phospholipids complexes technology. The effects of preparative parameters, such as emulsification time, stirring velocity and ultrasound time, on mean diameter, span of dispersity, entrapment efficiency and overall desirability were investigated. The three dimensional response surface graphs were produced by second-order polynomial and liner equation, which predict the optimal experiment conditions. All response variables were found to be greatly dependent on three independent variables. Second-order polynomial equations were fitter than liner equations for this study. The optimal emulsification time, stirring velocity and ultrasound time was 15 min, 2 000 r x min(-1), 30 min, respectively. The mean diameter, span of dispersity, entrapment efficiency, drug content and zeta potential of emulsions prepared by the method were 228.23 nm, 0.628 4, 84. 32%, 9.98 mg x mL(-1), - 29.03 mV, respectively. Puerarin submicron emulsion was prepared by the optimized preparation method. The narrow particle diameter distribution, high envelopment efficacy and good stability were obtained. The physicochemical properties were suitable for the requirement of the intravenous emulsion.
Emulsions ; Isoflavones ; administration & dosage ; chemistry ; Particle Size

Objective Using substituted benzyl piperazine as the raw material to design and synthesize new piperazine deriva-tives with protein tyrosine kinase(PTK)inhibitory activity. Methods Benzoic acid was used as starting compound to synthesize a key intermediate,2-chloroethyl benzoate,and the target compounds were synthesized by further reaction of the key intermediate with different substituted benzyl piperazine derivatives.Enzyme-linked immunosorbent assay(ELISA)was used to test the PTK inhibitory activity of the compounds.Results Fifteen new compounds were synthesized and their structures were verified by IR,1H NMR,MS, and elemental analysis.The PTK inhibitory activity of 3h and 3o was stronger than that of the other compounds.Conclusion The syn-thetic method is simple,and the raw materials are cheap and readily available.Compounds 3h and 3o showed relatively higher PTK in-hibitory activities.

Objective To study the routine examination of Aconitum sinomontanum Nakai and determine the contents of lappacontine and ranaconitine; To provide basis for establishing the quality standard of Aconitum sinomontanum Nakai.Methods Aconitum sinomontanum Nakai were collected from different areas.A method of TLC was used for qualitative discrimination. The methods in the Chinese Pharmacopoeia were adopted for the determination of moisture content, ash content and extractives. Determination of lappacontine and ranaconitine were performed by HPLC. Results The TLC showed that the spots were clear and the separation was good. Individual provisional standards:the moisture,total ash and acid-insoluble ash content of Aconitum sinomontanum Nakai were not more than 11.0%, 12.0%, and 7.0%, respectively; water soluble and alcohol soluble extractives were not less than 18.2% and 10.6%,respectively.The content of ranaconitine and lappacontine in Aconitum sinomontanum Nakai were not less than 0.125% and 0.815%, respectively. Conclusion The method established by the study is accurate and reliable,and can be used for quality evaluation of Aconitum sinomontanum Nakai.