OBJECTIVE:To establish RP-HPLC method for the determination of betahistine in human plasma.METHODS:The separation of betahistine was performed on C18 column with a detection wavelength of 261nm.the mobile phase was composed of 0.05mol/L ammonium acetate-acetonitrile-0.3mol/L sodium lauryl sulphate (60∶40∶1.5)with a flow rate of 0.5ml/min.RESULTS:The linear range of betahistine was 24～480ng/ml,the recovery rate was 81.75%～87.99%with the intra-day RSD at 0.60%～4.82%and inter-day RSD at 4.78%～12.15%.CONCLUSION:This method developed in the present study is simple,fast,accurate and reproducible,which can be used as the pharmacokinetic study for betahistine in human body.

Acupuncture ; Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Hiccup ; therapy ; Humans ; Male ; Middle Aged

Thyroid-stimulating antibodies were determined with cyclic AMP accumulation induced by unfractionaled serum using cultured human thyroid cells. Thyroid-stimulating antibodies were found in 83.3% (25/30) of patients with Graves' disease without treatment, 43.8% (7/16) of patients with Graves' disease during antithyroid treatment, 4% (1/25) of normal people and none of patients with simple goitre or thyroid adenoma. The results indicate that determination of thyroid-stimulating antibodies using this method is sensitive and characteristic in patients with Graves' disease.

AAV-ITR gene expression mini vector is a double-strand or single-strand DNA that only contains inverted terminal repeats of adeno-associated virus, cis-elements and gene of interest and does not contain any other foreign DNA sequences. We prepared Bac-ITR-EGFP and Bac-inrep. Spodoptera frugiperda cells were infected with Bac-ITR-EGFP (P3) and Bac-inrep (P3). Up to 100 μg of AAV-ITR-EGFP gene expression mini vectors were extracted from 2 x 10(7) cells of Sf9 72 h after infection. The gel electrophoresis analysis shows that most forms of AAV-ITR-EGFP gene expression mini vector were monomer and dimer. The mini vector expression efficacy was examined in vitro with HEK 293T cells. The EGFP expression was observed at 24 h after transfection, and the positive ratio reached 65% at 48 h after transfection.
Animals ; Baculoviridae ; DNA, Single-Stranded ; Dependovirus ; Gene Expression ; Genetic Vectors ; HEK293 Cells ; Humans ; Sf9 Cells ; Terminal Repeat Sequences ; Transfection

ObjectiveTo evaluate the effects of ventilator circuit change frequency on ventilatorassociated pneumonia (VAP).MethodsMeta-analysis of effects of ventilator circuit change frequency on VAP was conducted with study-level data from 1995 to 2010 in Pubmed,Embase,Web of Science databases.ResultsNine articles were included (sample size:20 326 mechanically ventilated patients).Analysis of six articles showed that the incidence of VAP in ventilator circuit change every 2 or 3 days was 4.05％,while 3.65％ in ventilator circuit change every 7 days.Compared with change ventilator circuit every 2 or 3 days,the risk ratio (RR) of VAP in weekly changes was 0.77 [0.54,1.09] ( P =0.14 ).Analysis three articles showed that compared to ventilator circuit change every 7 days with 15.89％ incidence of VAP,the incidence of VAP in circuit change more than 14 days was 14.9％,and RR was 0.98 [0.69,1.39](P =0.91 ).ConclusionsRegular ventilator circuit change frequency in various intervals can't difference in the incidence of VAP in mechanical ventilation patients.

Objective To investigate the change of the content of suppressor of cytokine signaling (SOCS-1) in the liver of septic mice and its working mechanism.Methods Adopted Cecalligation and puncture (CLP) to create models of sepsis and divided randomly adult male BALB/c mice into 8 groups,including normal controlled group,sham-operated group,and the killed groups 2 hours,6 hours,12 hours,24 hours and 48 hours after operation.After extracting the RNA and protein from the liver tissue of the mouse groups,reverse transcription polymerase chain reaction (RT-PCR) was adopted to determine the relative content of SOCS-1 mRNA in the tissue,Western blot was adopted to determine the relative content of protein and the SPSS statistics software was adopted to calculate the correlation.Then observed the pathological change of liver tissues,and detected SOCS-1 protein expression by immunohistochemistry.Results After CLP suergery,the expression of SOCS-1 on gene degree in the liver and the expression of SOCS1 on protein degree in the liver increased rapidly at the 6th hour ( P ＜ 0.05 ),with the former reaching peak ( P ＜ 0.05 ) at the 24th hour and the latter remaining high all the time.There were pathological changes such as fatty degeneration and necrosis in the septic liver tissue,hepatic SOCS-1 protein expression could be detected by immunohistochemistry.Conclusions CLP induced sepsis could lead to the increase of the expression of SOCS1 in the liver.

Objective To observe the antihypertensive effect of indapamide combined with enalapril on spontaneously hypertensive rats(SHRs).Methods Forty SHRs were randomly divided into 4 groups: control,indapamide,enalapril,and indapamide+enalapril(n=10 in each group).Medicine in varied doses was given to rats by intragastric administration.Variations of weight,heart rate and blood pressure were measured. Results Varied doses of medicine did not exert significant effects to the weight and heart rate of SHRs during and after the administration.In indapamide+enalapril group,the pressure of SHRs was significantly decreased with varied doses compared to that before the administration(P

Objective To investigate the expression of human bone morphogenetic protein-7(hBMP-7)in rat bone malTOW stromal cells(BMSC)with a recombinant adenovirai vector carrying the hBMP-7 gene(Ad-hBMP-7) and study the effecta of Ad-hBMP-7 transfection on BMSC difierentiation.in order to explore the possibility for hBMP-7 gene therpy.Methods The rat BMSC cultured in vitro.They were divided into 3 groups:untreated group,Ad-hBMP-7 and Ad-GFP transduced treated group.The rat BMSC were transfected by Ad-hBMP-7 and Ad-GFP. The expression of hBMP-7 was detected by RT-PCR and Western blot analysis.and the alkaline phosphatage (ALP)activity of the BMSC was observed.ResulIs In the Ad-hBMP-7 transduced treated group.hBMP-7 mRNA expression wag manifested detected by RT-PCR(470 bp),Westem blot analysis demonstrated that these cells indeed produced the hBMP-7 protein(Mr.15×103);10 days after transduction treatment,most of the BMSC were had brown black particles stained positively by ALP activity.But in Ad-GFP transduced treated group and untreated group they did not.Conclusions Ad-hBMP-7 could efficiently transfect BMSC and promote the conversion to osteoblast.The expression of hBMP-7 in rat BMSC provides a basis for hBMP-7 gene therapv.

Background The retina microglia play a eliminating effect on apoptotie cells in the neural retinal layer of normal rats during postnatal development.Milk fat globule epidermal growth factor 8(MFG.E8)can combine specifically with phosphatidylinositol serine of the surface of apoptotie cells and enhance macrophage phagoeytosis of apoptotic cells.Objective Present study was to evaluate the localization and expression of MFG-E8 and its relevant cytokines in the neural retinal layer of normal rats during postnatal development Methods Normal royal college of surgeon(RCS)rats were divided into P0,P3,P7,Pi4,P30,P45 groups according to their postnatal days,and the 30-day-old RCS rats(2 rats)served as controls.Double stain of M FG.E8 and microglial cells marker(CD11b)was performed by immunofluorescence.Expressions of MFG-E8,integrin β5,CD11b and interleukin-6(IL-6)mRNA in the neural retina were analyzed by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).The utilization of animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State and Science and Technology Commission.Results MFG-E8 and CD11b were positively co-expressed in retinal ganglion cell layer and external plexiform layer with the green fluorescence for FITC-labeled IgG and red fluorescence for cy3-labeled lgG respectively in normal adult rats.RT-PCR showed that the mRNA of MFG-E8,integrin 85,CD11b and IL-6 was detectable at P0 rats.The expression level of these eytokines began to rise fterward and reached peak value at P14 rats and then declined gradually,showing significant differences among different ages groups in various cytokines mRNA expression(all P<0.05).Conclusion MFG-E8 can be specifically expressed in the neural layer of retina microglia in RCS rat.

Toxic effects of L7 lyophilized powder of extracellular active components (L7-LPEAC), extracted from the Algae-lysing bacteria L7, on Chlorella pyrenoidosa were studied according to the changes of effective photosynthesis rate (EPR), the protein content, the chlorophyll a content and the MDA content of algae. The results showed that the growth of alga was promoted at low concentrations of L7-LPEAC (0.80 g/L, 1.25 g/L). The 96 h-EC50 and 120 h-EC50 upon Chlorella pyrenoidosa are 5.75 g/L and 2.55 g/L, respec tively. The chlorophyll a content increased firstly and then decreased at high concentrations of L7-LPEAC (≥2 g/L), so did the protein content. Compared with the control group, there is a significant statistics diff erence (P