Purpose　The aim is to immobilize the thrombin and to increase the stability of immobilized enzyme.Methods　Thrombin was immobilized on chitosan and crosslinked with glutaraldehyde, and the optimum conditions for immobilization and stability of immobilized enzyme were studied.Results　This preparation of immobilized thrombin was practicable,and the stability of this immobilized enzyme was obviously improved than that of thrombin.Conclution　The conditions for immobilization were good, and reliable.Stability of immobilized thrombin to light, high temperature,low temperature and observation on reserving at room temperature was good.

Aims Tocomparethepharmacokineticsof ephedra alkaloids,cinnamic acid and cinnamic alcohol in Ephedrae, Cinnamomi and Ephedrae-Cinnamomi herb couple through UPLC-MS/MS in rats respective-ly,and to investigate the effect of combination on phys-iologicaldisposition.Methods Plasmasampleswere collected at different times after oral administration of Ephedrae,Cinnamomi and Ephedrae-Cinnamomi herb couple extracts.The concentrations of ephedra alka-loids,cinnamic acid and cinnamic alcohol in plasma samples were determined by UPLC-MS/MS.DAS 3. 0 was used to calculate pharmacokinetic parameters.The differences of samples in two groups were conducted with univariate statistical analysis using SPSS 13. 0. Results ComparedwithEphedraegroup,theCmaxof norephedrine hydrochloride, norpseudoephedrine hydrochloride,ephedrine hydrochloride,pseudoephed-rine hydrochloride,and methylephedrine hydrochloride in Ephedrae-Cinnamomi herb couple group were signif-icantly greater (P<0. 05 );the AUC0-t of norpseudo-ephedrine hydrochloride was significantly greater (P<0. 05 );the MRT0-t of norephedrine hydrochloride, phedrine hydrochloride, pseudoephedrine hydrochlo-ride,and methylephedrine hydrochloride were signifi-cantly less (P <0. 05 );the T1/2z of norephedrine hydrochloride,phedrine hydrochloride,and methyl-ephedrine hydrochloride were significantly less (P <0. 05 ).The AUC0-t and MRT0-t of cinnamic acid and cinnamic alcohol were significantly greater than those inCinnamomigroup(P<0.05).Conclusion The combination of Ephedrae and Cinnamomi improves the absorption concentration of five ephedra alkaloids, Slows down the elimination of norephedrine hydrochlo-ride,phedrine hydrochloride,pseudoephedrine hydro-chloride,and methylephedrine hydrochloride,and in-creases the bioavailability of cinnamic acid and cin-namic alcohol.

Objective Through clinical observation and research with persistent atrial fibrillation in patients with blood-brain natriuretic peptide ( BNP) level change detection renin, angiotensin Ⅱ, aldosterone, analyze their data, investigate persistent atrial fibrillation ( PAF ) serum brain natriuretic hormone levels change and the relationship between the renin-angiotensin-aldosterone system (RAAS).Methods 80 patients with coronary artery disease in our hospital from December 2013 to October 2015 were selected, randomly divided into experimental principle PAF group (40 cases) and control group ( patients in sinus rhythm,with 40 cases) , BNP, renin, angiotensinⅡ, aldosterone levels were measured in two groups, and left atrial diameter (LAD) measurements.Results Under the same condition of cardiac function, the blood BNP concentration in PAF group was higher than that in control group (P <0.05).The results of the linear regression analysis showed that the correlation coefficient of blood BNP level and AngII concentration in PAF group was 0.301, the correlation coefficient of blood BNP level and LAD indicators in PAF group was 0.373, the concentrations of BNP and RAAS were positively correlated with LVMI, the differences were statistically significant (P<0.05), the blood BNP level was closely related to the level of RAAS in the PAF group (P<0.05).Conclusion Blood BNP concentration was significantly higher in patients with PAF and blood BNP concentration is closely related with the RAAS, LAD indicators, and a positive correlation.

Objective To study the serum levels of proinsulin(PI),insulin(INS),proamylin(PA) and amylin(AMY) in abnormal glucose metabolism individuals and to explore the relationship between PI and insulin resistance(IR) and between PA and islet beta cell function.Methods Totally 79 subjects who received Oral glucose tolerance test(OGTT) and insulin release test in our department from March to May 2008 were divided into 4 groups according to the results of OGTT,that is,normal group,impaired glucose regulation(IGR) group,T2DM 1[fasting blood-glucose(FBG)15 mmol/L) group.All subjects underwent examination of anthropometry and serum bio-chemical indicators,HOMA-IR and HOMA-B were calculated.Results No significant difference was found in the serum levels of PI or AMY among the 4 groups,but the level of PA,PI/INS and PA/AMY among the 4 groups did have significant differences(P

Objective:To collect and evaluate the information of literature on perimenopausal syndrome.Methods:The computer retrieval method supplemented by manual inspection of literature.Results:590 relevant documents were retrieved and among which 129 were comprehensively studied.Conclusion:The quality of the literature should be improved though there was an annual quantitive increase of the literature on perimenopausal syndrome in TCM.Great efforts should be made in this aspect.

OBJECTIVE To study the prevalence,fungal species,therapy and prevention of invasive fungal infections in our Affiliated Hospital.METHODS From Jan 2004 to Dec 2005,all patients with positive results for fungal culture of blood and sterile fluid were reviewed retrospectively.The in vitro activity of antifungal agents against fungal isolates was determined.RESULTS Eighty-eight positive patients were collected in this analysis based on the diagnosis of invasive fungal infection in our hospital for two years,from them the majority were diagnosed as candidal infections(n=72,81.8%),the others were Cryptococcus neoformans(n=8,9.1%),Aspergillus(n=5,5.7%) and other different fungal infections.The most common was C.albicans(n=48,66.7%),C.glabrata(n=12,16.7%),and C.tropicalis(n=8,11.1%),C.parapsilosis and C.krusei were both the same results(n=2,2.8%).CONCLUSIONS Infections caused by Candida are the most common invasive fungal ones in our hospital,and C.albicans is the most common one.Fluconazole remains effective for treament of candidal infections,especially those caused by C.albicans.

To simultaneously determine the contents and explore the pharmacokinetics of ferulic acid and paeoniflorin by high performance liquid chromatography (HPLC) after oral administration of Modified Xiao-yao Decoction (MXYD), a compound of traditional Chinese herbal medicine.

Objective To construct a luciferase reporter plasmid containing interferon regulatory factor 3(IRF-3)human gene promoter and to evaluate promoter activity in human embryonic kidney(HEK)-293 cells.Methods The 1 000 bp fragment was amplified by PCR with human genomic DNA as a template and was directionally cloned into pGL3-basic multiple cloning sites to construct the luciferase repor-ter plasmid pGL3-pIRF-3.Transfection of HEK-293 cells with the promoter-driven lucife-rase construct was performed to induce lucife-rase gene expression and calculate the relative luciferase activity unit(RLU).Promoter sequence of 1 000 bp upstream of transcription initiation site of IRF-3 was analyzed by using Promoter 2.0 Prediction software.Results DNA sequencing and restriction endonuclease analysis verified the successful construction of the plasmid pGL3-pIRF-3.This IRF-3 promoter exhibited a strong promoter activity with an increase of 42.2-fold of RLU in HEK-293 cells when compared with pGL-3 basic vector.The transfection experiment confirmed that the levels of its activation were significantly higher than that in controls in HEK-293 cells.Function analysis of IRF-3 promoter disclosed seve-ral GATA-1 and specific protein 1(Sp1) sites and E2F in minimal promoter region.Conclusion The plasmid pGL3-pIRF-3 promoter is successfully constructed and has a strong basal promoter activity in HEK-293 cells.

Objective To estimate the uncertainty of measurement in detecting of hepatitis B virus DNA(HBV DNA)by method of fluorescence quantitative polymerase chain reaction(FQ-PCR),and discuss the application value.Methods The process of the detection of HBV DNA by FQ-PCR was analysed to confirm and simplify the sources of uncertainties of measurement,which were obtained by disposing the data of methodology validation,internal quality control(type A evaluation of uncertainty)and external quality assessment(type B evaluation of uncertainty);combined uncertainty and expanded uncertainty were obtained by statistical methods.Results The main sources of uncertainties of measurement were:precision within laboratory,precision between laboratory,method bias.The expanded uncertainty of HBV DNA by FQ-PCR was U=0.62(k=1.96,n=2).The uncertainty caused by method bias was found mostly.Conclusion Expanded uncertainty can be compared in different results of HBV DNA by FQ-PCR,and it provides guide significance for observing the cure effect of anti-HBV and choosing the concentration of quality control.

OBJECTIVE: To probe the correlation between the postmortem interval (PMI) and the DNA degradation of costicartilage and dental pulp cells in human being after death, and to seek a new method for estimating PMI. METHODS: The image cytometry was used to measure the DNA degradation under different ambient temperatures (30-35 degrees C, 15-20 degrees C) in 0-15 days after death. RESULTS: The average DNA content of two kinds of tissue was degradated with the prolongation of PMI. But there was a plateau period of 0-4 days for dental pulp cells of human being in 15-20 degrees C. There was a high negative correlativity P<0.01 between the average DNA content and PMI. CONCLUSION PMI could be estimated accurately according to the DNA degradation of costicartilage and dental pulp cells in human being after death.
Adolescent ; Adult ; Autopsy ; Cartilage/metabolism* ; DNA/metabolism* ; Dental Pulp/cytology* ; Female ; Forensic Pathology/methods* ; Humans ; Image Processing, Computer-Assisted/methods* ; Male ; Middle Aged ; Postmortem Changes ; Ribs/metabolism* ; Temperature ; Time Factors ; Young Adult