To construct, express and purify fusion protein containing HBcAg and HBsAg preS1 epitope peptide for the purpose of investigating a novel HBV vaccine with both prophylactic and therapeutic functions. Using DNA recombinant technology, prokaryotic expression plasmid pBTcs1 expressing HBcAg and HBsAg pre-S1 epitope peptide fusion protein was constructed. After expressed in E.coli. HB101, the production BTcs1 was purified by sucrose density gradient ultracentrifugation and identified by SDS-PAGE, SEC, Western-blot and electron microscope. The results indicated that expression plasmid pBTcs1 was constructed successfully, and 20~25 mg purified BTcs1 fusion protein was obtained from 1L LB culture. Result of DOT-BLOT indicated that the distribution of BTcs1 was mainly in 30~50% sucrose, the purity of BTcs1 was greater than 95% by SDS-PAGE and SEC analysis. BTcs1 could probe with specific antibodies at 28 kDa by Western-blot, BTcs1 could also self assemble VLP by electron microscope analysis, its diameter was 30~34 nm approximately. The present study lay a foundation for further research functions and applications of BTcs1.

1:16) of the relative serum antibody has been observed in 80%, while none of this high level was observed in patients with Echovirus-25 and adenovirus dermatitis (

Adolescent ; Female ; Humans ; Mucocutaneous Lymph Node Syndrome ; complications ; Uveitis, Anterior ; etiology

AIMTo determine clonidine in rabbit plasma by LC-MS.

METHODSThe LC-MS system consisted of Waters Alliance 2790 HPLC and Micromass ZQ-4000 MS. The HPLC was performed by using XTerra C18 (150 mm x 2.1 mm ID, 5 microm). The mobile phase, consisting of acetonitrile/ammonium hydrogen carbonate solution, was maintained to a flow-rate of 0.2 mL x min(-1) and the linear gradient elution was adopted. Mass spectrum was obtained by using electrospray ionization interface and the m/z of SIM was 230.

RESULTSThe average recovery was high and the method was reproducible. The calibration curve showed good linearity in the range of 1 - 80 microg x L(-1), the lowest limit of detection was 0.05 microg x L(-1). The Cmax, AUC0-t, and Tmax value of the pharmacokinetics parameter were (27 +/- 9) microg x L(-1), (5,352 +/- 1,121) microg x L(-1), (79 +/- 17) h.

CONCLUSIONThe results demonstrated that the method had high sensitivity, good selectivity, accuracy and precision. It is used to determine the clonidine concentration in plasma. The transdermal patch can deliver clonidine to the surface of rabbit skin stably for periods of up to 1 week after a single application.

Administration, Cutaneous ; Animals ; Antihypertensive Agents ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Clonidine ; administration & dosage ; blood ; pharmacokinetics ; Rabbits ; Spectrometry, Mass, Electrospray Ionization ; methods

OBJECTIVETo evaluate the effects of paroxetine on protein kinase PKA, PKC and CaMKII activities in different brain regions in a rat model of depression.

METHODSThirty-six adult male SD rats were randomized into 6 groups, including one control group (I) and 5 groups of depression model established by forcing the rats to swim for 4 weeks. The 5 depression groups received no treatment (II) or were treated with paroxetine at a single dose (III), for a week (IV), 2 weeks (V) or 4 weeks (VI). The radioactivity of PKA, PKC and CaMKII in the hippocampus and prefrontal cortex was quantitatively measured using a liquid scintillation counter.

RESULTSIn the rat hippocampus, PKA and CaMKII activities were significantly lower in groups II, III, IV, and V than in groups I and VI (P<0.01 or P<0.05), but comparable between groups VI and I (P>0.05). PKC activity was significantly lower in group II than in group I (P<0.01), but showed no significant difference between the paroxetine-treated groups and group I (P>0.05). In the prefrontal cortex, the activity of PKA in groups I, II, III, and IV was similar (P>0.05), but all significantly lower than that in groups V and VI (P<0.01). PKC activity was significantly higher in groups II and III than that in group I and other paroxetine-treated groups (P<0.01), and similar between groups IV and I (P>0.05); groups V and VI had significantly lower PKC activity than group I (P<0.01). Group I had the highest CaMKII activity among the groups (P<0.01).

CONCLUSIONChronic administration of paroxetine can reverse chronic stress-induced inhibition of PKA, PKC and CaMKII activity in rat hippocampus, while the effects of paroxetine on the protein kinases can be more complex in prefrontal cortex.

Animals ; Brain ; drug effects ; enzymology ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Depression ; enzymology ; Disease Models, Animal ; Hippocampus ; drug effects ; enzymology ; Male ; Paroxetine ; pharmacology ; Protein Kinase C ; metabolism ; Random Allocation ; Rats

Objective To evaluate the relationship between interleukin-18(IL-18) and insulin resistance, measured by euglycemic hyperinsulinemic clamp, in women with polycystic ovary syndrome (PCOS). Methods Forty-two young women with PCOS and 38 age-and body mass index (BMI)- matched control women were recruited in this study. A complete hormonal assay was performed and serum IL-18 level was determined in each subject. And euglycemic hyperinsulinemic clamp test was completed in 41 of the above subjects. Results Serum IL-18 levels were increased in the PCOS women, as compared with the control (P=0.033). When all subjects were divided into lean and obese groups, the IL-18 levels were slightly increased in the obese subjects (P=0.902). IL-18 levels were negatively correlated with all the clamp parameters [mean glucose infusion rate (M), M-to-insulin ratio (M/I) and glucose metabolic clearance rate (MCRg), r =-0.419,-0.396,and-0.405,P=0.006,0.010 and 0.009 respectively], but were positively associated with HOMA-β index(r=0.334, P=O.035). Conclusion Serum IL-18 level was significantly increased in PCOS women and was strongly associated with the parameters obtained from the euglycemic hyperinsulinemic clamp, indicating that IL-18 may be an important mediator between inflammation and insulin resistance.

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.

The purpose of this study was to investigate the preparation and characteristics of curcumin phospholipid complex, including the effects of reaction time, reaction solvent, reaction concentration and reaction temperature. Preparation technology resulted in that 0.5 g curcumin and 10 g soy phospholipid dissolved in 100 mL anhydrous alcohol, were stirred 1 h in 50 degrees C waterbath, then steamed alcohol in decompression, collected solid residue and vacuum dried for 12 h. The physicochemical properties for the new complex including IR spectrometer, mass spectrograph and HNMR equipment were detected. As a result, the formation of the complex is based on the reaction between phospholipid polar group rounding phosphorus atom and curcumin. This result gave the evidence for the formation mechanism of phospholipid complex.
Curcuma ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Phospholipids ; chemistry

OBJECTIVETo use a new kind of fixing material, i.e. Sol-Gel organic-inorganic hybridized material to immobilize bacterium to detect Biochemical oxygen demand quickly.

METHODSThe biosensor was fabricated using a thin film in which Hansenula anomala was immobilized by sol-gel and an oxygen electrode. The optimum measurement for biochemical oxygen demand was at pH 7.0; 28 degrees C; response time 3 - 12 min. Pure organic compound, sewage and rate of recovery were detected with the biosensor.

RESULTSIt shows that the BOD biosensor can be used to detect many organic compounds such as amino acid, glucide. It is suitable to monitor sewage and industrial waste water which has low level alcohols and phenols. The microbial membrane can work 3 months and remain its 70% activity. It is measured that the rate of recovery of BOD is between 90% to 105% in sewage.

CONCLUSIONThe study confirmed the effectiveness and usefulness of BOD sensor, which is quick, convenient, low cost and reliable with little interference.

Bacteria ; Biosensing Techniques ; instrumentation ; Cells, Immobilized ; Gels ; Membranes, Artificial ; Nylons ; Oxygen ; analysis ; Sewage ; analysis ; microbiology

Objective To explore the possible correlation between serum highly sensitive C reactive protein (hs CRP) and blood glucose, insulin, lipids ,insulin sensitivity index (SI),acute insulin response(AIR), and adiponectin in subjects with impaired glucose tolerance (IGT) or newly diagnosed type 2 diabetes mellitus(DM). Methods SI and AIR were assessed by the reduced sample number of Bergman′s minimal model method by intravenous glucose tolerance test in subjects. Meanwhile body mass index (BMI), waist hip ratio (WHR), the serum lipid profile, hs CRP, adiponectin levels were measured. Results Compared with normal control (NC) group[SI(6.6?2.4) 10 -4 (min?mU/L) -1 ,adiponectin7.77(6.35 10.70 mg/L),hs CRP0.40(0.21 1.67mg/L)], the SI and serum adiponectin in IGT group [(1.5?1.1) 10 -4 (min?mU/L) -1 , 4.29(3.59 6.22 mg/L) respectively] and type 2 DM group [(1.5?1.0)?10 -4 (min?mU/L) -1 , 3.46(2.37 4.72 mg/L) respectively] were significantly decreased (all P