Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.

OBJECTIVETo compare the efficacy of two surgical techniques for controllong nasal width after Le Fort I osteotomy.

METHODSFifty-five patients who received the Le Fort I osteotomy have been included in this study. They were randomly divided into 2 groups. The experimental group received extraoral ABS, and the control group received traditional intraoral ABS. 3D photos of the patient's face were taken before operation and at postoperative 3 months. Alar width was measured on the 3D photos. Data was reported as means and standard deviations, and statistic analysis was done by using student t test.

RESULTSCompared with presurgical data, G. lat-G. lat increased by (2.66 +/- 1.47) mm, Al-Al increased by (2.20 +/- 1.22) mm and Sbal-Sbal increased by (1.30 +/- 1.33) mm in experimental group. G. lat-G. lat increased by (1.38 +/- 1.29) mm, Al-Al increased by (1.06 +/- 0.95) mm and Sbal-Sbal increased by (0.36 +/- 1.33) mm in the control group. There was significant difference between two groups.

CONCLUSIONSThe surgical technique of ABS is the most important factor for determining the postoperative alar width. Both techniques have better effect on the Sbal-Sbal width control than the G. lat-G. lat and Al-Al width control. Traditional intraoral ABS can more effectively control the alar width. Both techniques cannot completely control the alar base widening after Le Fort I osteotomy.

Face ; Humans ; Nose ; anatomy & histology ; Nose Deformities, Acquired ; surgery ; Osteotomy, Le Fort ; adverse effects ; Photography

Animals ; Dose-Response Relationship, Drug ; Male ; Mice ; Mice, Inbred Strains ; Nickel ; toxicity ; Oxidative Stress ; physiology ; Testis ; drug effects ; metabolism

Paracetamol was used as a model drug in this study to investigate the synergetic effects of lipid coating and beta-cyclodextrin (beta-CD) inclusion for masking the bitter taste of poorly soluble drugs. To control the concentration as low as possible of the free drug which produced a bitter taste, a kinetic model was established to calculate the drug distribution theoretically among the free drug in medium, lipid coated particles and molecular inclusion on the basis of the preparation and characterization of the lipid microspheres, so as to select the proper amount of beta-CD. Finally, the synergetic drug delivery systems were prepared and characterized by 1H nuclear magnetic resonance (1H NMR), molecular simulation and the electronic tongue. As a result, the drug release rate constant (k) of the lipid microspheres coated with octadecanol was determined as 0.001 270 s(-1). Then, the synergetic drug delivery systems were prepared with the ratio of 6.74 : 1 (w/w) for beta-CD and paracetamol. The chemical shift values for the fingerprint peaks of paracetamol all increased and hydrogen bonds were formed between the oxygen on the phenolic hydroxyl group, the nitrogen on the imino in paracetamol and the hydrogens on the hydroxyl groups in beta-CD. The results tested by the electronic tongue indicated that the paracetamol, lipid microspheres, beta-CD inclusion and their mixture showed different taste characteristics, with the bitterness order of the synergetic drug delivery systems approximately lipid microspheres < beta-CD inclusion < paracetamol, which confirmed the synergetic taste masking effects of lipid coating and beta-CD molecular inclusion. In summary, the synergetic taste masking was jointly achieved through the retard of the drug release by the lipid coating and the inclusion of the free paracetamol by beta-CD through hydrogen bonds.
Acetaminophen ; administration & dosage ; chemistry ; Administration, Oral ; Drug Delivery Systems ; Electrical Equipment and Supplies ; Electrochemical Techniques ; instrumentation ; methods ; Hydrogen Bonding ; Kinetics ; Lipids ; chemistry ; Microspheres ; Solubility ; Taste ; drug effects ; beta-Cyclodextrins ; chemistry

Objective To study the effect of the petroleum ether fraction of ethanol extracts of Polyrhachisvicina Roger with antigout activity on liver microsomal cytochrome P450 in rats.Methods Healthy male SD rats were randomly divided into six groups (3 rats of each group):three groups of the petroleum ether fraction of ethanol extracts of Polyrhachisvi-cina Roger ( test groups, the low dose group, the middle dose group and the high dose group with equivalent to 1, 2, 4 g? kg -1, respectively), two positive control groups and one blank control group.The rats in the low dose, the middle dose and the high dose test groups were adminis-tered daily by gavage at corresponding dose of the petroleum ether frac-tion of ethanol extracts of Polyrhachisvicina Roger for 10 consecutive days.The rats in positive control groups were treated by dexamethasone ( 100 mg? kg-1? d-1 , ip, daily for 4 days ) or phenobarbital ( 80 mg? kg-1? d-1 , ip, daily for 3 days ) .Blank control rats received equivalent volume of sterile normal saline daily by gavage for 10 consecu-tive days.The levels of CYP450 activity, mRNA, protein in rat liver mi-crosome were analyzed by LC/MS/MS or RP-HPLC-FLD, RT-PCR, Western blot, respectively.Results CYP1A2 activity, protein expression and mRNA levels were increased signifi-cantly in a dose-dependent manner with the petroleum ether fraction of ethanol extracts of Polyrhachisvicina Roger at low, middle and high dose, respectively.Conclusion The petroleum ether fraction of ethanol extracts of Polyrhachis-vicina Roger can induce CYP1A2 in rats, suggesting the potential of drug-drug interactions between the petroleum ether fraction of ethanol extracts of Polyrhachisvicina Roger and the inducers, inhibitors, or substrates of CYP1A2.

Objective To explore the expression of P -glycoprotein ( P-gp) in rat liver during acute inflammation ( AI ) and the possible mechanism involved.Methods Twenty rats were randomly divided into two groups:model group (n=10) and control group (n=10).The rats in model group were treated by a single 5 mg? kg -1 intraperitoneal injec-tion of lipopolysaccharide ( LPS).Control rats received equivalent injec-tion of sterile normal saline.The levels of P-gp, ubiquitinated P-gp and 26 S proteasome activity in both groups were analyzed by Western blot, immunoprecipitation and fluorescence detection, respectively. Results Compared to the control groups, ubiquitination levels of liver P-gp and 26S proteasome activity in model groups were significantly in-creased ( P<0.01 ) , accompanied by an decrease of liver P-gp protein expression level. Conclusion The participation of the ubiquitin -proteasome system in down-regulation of liver P-gp expression levels under AI conditions.

Oroxylum indicum was a traditional Chinese medicine. In order to study the chemical constituents from the seed of O. indicum, the chemical constituents of 80% methanol extract of seeds of O. indicum were subjected to chromatography on silica gel, Sephadex LH-20, and preparative HPLC, leading to the isolation of eleven compounds. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data as oroxin B (1), chrysin (2), baicalein (3), neglectein (4), quercetin-3-O-β-D-galactopy ranoside (5), quercetin-7-O-β-D-glucopyranoside (6), 2α,3β-dihydroxylluPeol (7), lupeol (8), rengyol (9), β-sitostero (10), and stigmasterol (11). Among them, compound 5 were firstly obtained from O. indicum.
Bignoniaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Seeds ; chemistry

Paracetamol was used as a model drug in this study to investigate the synergetic effects of lipid coating and beta-cyclodextrin (beta-CD) inclusion for masking the bitter taste of poorly soluble drugs. To control the concentration as low as possible of the free drug which produced a bitter taste, a kinetic model was established to calculate the drug distribution theoretically among the free drug in medium, lipid coated particles and molecular inclusion on the basis of the preparation and characterization of the lipid microspheres, so as to select the proper amount of beta-CD. Finally, the synergetic drug delivery systems were prepared and characterized by 1H nuclear magnetic resonance (1H NMR), molecular simulation and the electronic tongue. As a result, the drug release rate constant (k) of the lipid microspheres coated with octadecanol was determined as 0.001 270 s(-1). Then, the synergetic drug delivery systems were prepared with the ratio of 6.74 : 1 (w/w) for beta-CD and paracetamol. The chemical shift values for the fingerprint peaks of paracetamol all increased and hydrogen bonds were formed between the oxygen on the phenolic hydroxyl group, the nitrogen on the imino in paracetamol and the hydrogens on the hydroxyl groups in beta-CD. The results tested by the electronic tongue indicated that the paracetamol, lipid microspheres, beta-CD inclusion and their mixture showed different taste characteristics, with the bitterness order of the synergetic drug delivery systems approximately lipid microspheres < beta-CD inclusion < paracetamol, which confirmed the synergetic taste masking effects of lipid coating and beta-CD molecular inclusion. In summary, the synergetic taste masking was jointly achieved through the retard of the drug release by the lipid coating and the inclusion of the free paracetamol by beta-CD through hydrogen bonds.

OBJECTIVETo study the mechanism of the genital system damage by nickel sulfate in male rats in order to provide the laboratory theoretical evidence for the prevention and cure of nickel genital toxicity.

METHODSThree groups of rats were injected intraperitoneally with nickel sulfate at dose of 1.25, 2.50, 5.00 mg/kg respectively for two weeks. The content of testicle nickel and blood serum testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH) were assessed with atomic absorption spectrum and radioimmuno-assay, meanwhile the activity of nitric oxide synthase (NOS) and the content of nitric oxide (NO) were measured by enzyme method.

RESULTSThe contents of testicle nickel [(0.22 +/- 0.03), (0.34 +/- 0.04), (0.41 +/- 0.02) micro g/g respectively] were increased, but the content of T, TSH, LH in blood serum were reduced; the activities of NOS in testicle tissue [(33.65 +/- 2.93), (26.53 +/- 9.52), (10.20 +/- 2.74) U/g respectively] were inhibited by nickel sulfate and the contents of NO [(0.26 +/- 0.03), (0.18 +/- 0.05), (0.15 +/- 0.02) mmol/g respectively] were decreased (P < 0.01).

CONCLUSIONNickel-induced genital system injury of male rats may be related to the decrease in the contents of T, TSH, LH, and the inhibition on NOS, as well as the fall of NO content.

Animals ; Follicle Stimulating Hormone ; blood ; Genitalia, Male ; drug effects ; metabolism ; pathology ; Luteinizing Hormone ; blood ; Male ; Nickel ; toxicity ; Nitric Oxide ; analysis ; Nitric Oxide Synthase ; analysis ; Radioimmunoassay ; Random Allocation ; Rats ; Rats, Wistar ; Testosterone ; blood

Adult ; Antigens, CD20 ; metabolism ; Cervical Vertebrae ; pathology ; Eosinophilic Granuloma ; complications ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Hodgkin Disease ; complications ; metabolism ; pathology ; surgery ; Humans ; Ki-1 Antigen ; metabolism ; Male ; Middle Aged ; Spinal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Thoracic Vertebrae ; pathology