Objective To investigate the biological function of SPA-PEI conjugate(staphylococcal protein A-polyethyleneimine cross-linker),which is one key component for construction of a novel antibody-targeted DNA delivery system.Methods The binding capacity of SPA-PEI conjugate with multiple sources of IgG was determined by enzyme-linked immunosorbent assay and neutralization inhibition assay.The binding capacity of SPA-PEI conjugate with DNA fragment was determined by DNA gel retardation assay,and its DNA condensing ability was measured by Ethidium bromide exclusion assay.Results SPA-PEI conjugate could bind well to many species-derived IgGs.SPA-PEI conjugate had no significant effect on the binding properties of SPA.SPAPEI conjugate could neutralize negative charges of the plasmid DNA or DzTi.Its DNA condensing ability was nearly same to that of free PEI,which suggested a excellent DNA condensing ability of the SPA-PEI conjugate.Conclusion SPA-PEI cross-linkers prepared by this project group maintained the biological activity of SPA and PEI.SPA-PEI cross-linkers could be used for the construction of a novel antibody-targeted non-virus DNA delivery system.

To improve the class effectiveness,different teaching methods were chosen based on the characteristics of pharmacy students,such as to stimulate the students’ interest,raise the independently ability,design the appropriate multimedia content and practice.

Aged ; Aged, 80 and over ; Coronary Artery Disease ; complications ; therapy ; Coronary Stenosis ; complications ; therapy ; Female ; Humans ; Male ; Middle Aged ; Percutaneous Coronary Intervention

Objective To study the relationship between sperm activity and the level of L-carnitine in seminal plasma of patients with idiopathic asthenospermia by HPLC.Methods Experimental group:idiopathic asthenospermia 50 cases,and the control group:male with pregnancy history 10 cases.The semen parameters (volume,sperm density,viability,motility),and L-carnitine level in seminal plasma were detected with high performance liquid chromatography method.Results There were significant difference between A and B group about sperm density and the percentage of progressive motile spermatozoa,and the level of L-carnitine(P＜0.01).Conclusions The decrease of L-carnitine level in patients with idiopathic asthenospermia,is closely related to the low sperm motility,which shows that L-carnitine can be used to treat idiopathic asthenospermia effectively.

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Using a UPLC-MS/MS (MRM) and cocktail probe substrates method, the metabolic fingerprint of the compatibility of Radix Aconite (RA) and Radix Paeoniae Alba (RPA) and its effect on CYP450 enzymes were investigated. These main CYP isoforms include CYP 1A2, CYP 2C, CYP 2E1, CYP 2D and CYP 3A. Compared with the inhibition effect of RA decoctions on CYP450 isoforms, their co-decoctions of RA and RPA with different proportions can decrease RA' inhibition on CYP3A, CYP2D, CYP2C and CYP1A2, but can not reduce RA' effect on CYP2E1. The metabolic fingerprints of RA decoction and co-decoctions with different proportions of RPA in CYP450 of rat liver were analyzed by UPLC-MS. Compared with the metabolic fingerprints of RA decoction, the intensity of diester-diterpenoid aconitum alkaloids decreased significantly, while the intensity of monoester-diterpenoid alkaloids significantly increased in the metabolic fingerprints of co-decoctions of RA and RPA. The results suggest that RA coadministration with RPA increased the degradation of toxic alkaloid and show the effect of toxicity reducing and efficacy enhancing.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Liver ; drug effects ; enzymology ; Metabolome ; Paeonia ; chemistry ; Rats ; Tandem Mass Spectrometry

Adolescent ; Adult ; Astrocytoma ; diagnosis ; surgery ; Brain Neoplasms ; diagnosis ; surgery ; Child ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Microsurgery ; methods ; Middle Aged ; Thalamic Diseases ; diagnosis ; surgery ; Thalamus ; surgery ; Treatment Outcome

OBJECTIVETo explore the effect of Yixintai Granule (YG) on mRNA and protein expression levels of AQP2 in renal medulla of chronic heart failure (CHF) rabbits.

METHODSCHF rat model was established by ear marginal vein injection of adriamycin. Successfully modeled rabbits were divided into the model group, the high (8.4 g/kg), middle (4.2 g/kg), and low dose (2.1 g/kg) YG group, and the Furosemide group (2 mg/kg). Besides, a normal control group was set up. Equal volume of physiological saline was administered to rabbits of the model group and the normal control group by gastrogavage. YG at different doses was administered to rabbits of the 3 YG groups by gastrogavage. The intervention lasted for 4 weeks, once per day. After treatment the urine volume and pathomorphological changes of renal medulla tissue were observed. mRNA and its protein expression levels of AQP2 were detected.

RESULTSCompared with the normal control group, the urine volume decreased significantly, mRNA and protein expression levels of renal medulla AQP2 increased significantly in the model group (all P < 0.01). Compared with the model group, the urine volume increased significantly, and mRNA and protein expression levels of renal medulla AQP2 decreased significantly in all medicated groups (all P < 0.01). Compared with the low dose YG group, the urine volume significantly increased and the mRNA expression level of renal medulla AQP2 significantly decreased in the middle and high dose YG groups (all P < 0.01). The expression level of AQP2 protein significantly decreased in the high dose YG group (P < 0.01). Pathological changes of the renal medulla was the most obviously seen in the model group. But they were alleviated to various degrees in all medicated groups. They were more obviously attenuated in the middle and high dose YG groups.

CONCLUSIONYG could improve CHF possibly through down-regulating mRNA and protein expression levels of AQP2 in renal medulla, and elevating the urine volume.

Animals ; Aquaporin 2 ; genetics ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Heart Failure ; drug therapy ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Rats, Sprague-Dawley

Objective To study the cyfluthrin resistance and potential mechanisms of Anopheles sinensis in Nanchang Chang-bei International Airport,Nanchang City,Jiangxi Province. Methods The resistance levels of the local An. sinensis were de-tected by WHO drug resistance bioassay. During the bioassay,the dying mosquitos were classed as sensitive mosquitos,and the survival ones were classed as resistant mosquitos. The P450 monooxygenase activity and glutathione S-transferase activity were detected and compared between the two groups. At the same time,the death time of each sensitive mosquito was recorded,and the correlations between the death time and the P450 monooxygenase activity and glutathione S-transferase activity were ana-lyzed,respectively. Results The bioassay mortality of the local An. sinensis was 59.5%. The differences of the P450 monooxy-genase activities among the resistant mosquitos,sensitive mosquitos and laboratory sensitive mosquitos had statistical signifi-cances(F=151.89,P<0.01),the resistant mosquitos>sensitive mosquitos>laboratory sensitive mosquitos. The differences of glutathione s-transferase activities among the three groups had no statistical significance(F=0.72,P=0.49). There existed positive correlation between the mosquito death time and the P450 monooxygenase activity,and the regression equation was y=79.479+1.512x with the correlation coefficient of 0.88,while there was no correlation between the mosquito death time and the glutathione S-transferaseactivity. Conclusion The An. sinensis in Nanchang Changbei International Airport has been resistant to cyfluthrin,and the promotion of P450 monooxygenase activity maybe one of the reasons for the resistance.

Objective To investigate the effect of neutral amino acid on preventing Parkinson disease.Methods Mice were injected with L-Valine,L-Pheylalanine,D-Valine or L-Lysine before or after paraquat administration,by which prakinsonian mouse model was constructed.The paraquat immunoreactivity was observed within nigral cell bodies.Then neurodegeneration and ?-synuclein aggregation were observed by immunohistochemistry and Western blot.Results Paraquat immunoreactivity was abolished by the administration of L-Valine,L-Pheylalanine before paraquat exposure.Pre-treatment with these two amino acids also protected the paraquat-induced loss of nigrostriatal dopaminergic cells and formation of thioflavine S-positive aggregates.In contrast, paraquat-induced toxicity was unaffected if animals were injected with these two amino acids after paraquat exposure or pre-treated with D-Valine or L-Lysine.Conclusions L-type neutral amino acids such as L Valine and L-Pheylalanine can prevent paraquat-induced neurodegeneration and a synuclein pathology through a competitive inhibition mechanism with stereospecificity in the central nervous system (CNS).Neutral amino acid could protect the dopaminergic neuron in substantia nigra and may prevent Parkinson disease.