Objective To search for a candidate DNA vaccine of Fasciola hepatica. Methods Using RT-PCR and digestion with Hind III and BamHI, Fasciola hepatica secreted cathepsin L1(FheCL1) cDNA was cloned into the expression vector pcDNA3.1. Results The cloning was successful, the cDNA sequence and its deduced amino acid sequence were analyzed. There was much difference between the cloned FheCL1 and the published one. But the first 20 residues of their amino acid sequences were the same. Conclusion The recombinant plasmid pcDNA3.1-FheCL1 may be a new type of candidate DNA vaccine candidate for Fasciola hepatica. It is possible that Fasciola hepatica presents different sub-species but their amino acid residues (1 to 20) encoded by FheCL1 might build up membrane spanning helix.

Objective To set up an experiment model on cellular level in order to investigate the effects of aconitine on Connexin43(Cx43)in cardiomyocytes of rat.Methods The cultured cardiomyocytes of rat were incubated with aconitine at 6 different concentrations of 0.25,0.5,0.75,1.0,1.5 and 2.0 ?mol/L.The changes of Cx43 phosphorylation status of each group were investigated by Western blot analysis and immunofluorescent microscopy techniques.Results The total amount of Cx43 had not changed in cardiomyocytes after aconitine incubation by western blot analysis,but it began to induce Cx43 dephosphorylation after incubation of the cultures with 0.5 ?mol/L aconitine and Cx43 underwent significant dephosphorylation when the concentration of aconitine elevated to 1.0 ?mol/L,and the dephosphorylation effects of 1.5 and 2.0 ?mol/L aconitine on Cx43 were similar to that of 1.0 ?mol/L aconitine.Quantitative immunofluorescent microscopy revealed that Ser368,one of the serine amino acid phosphorylation sites in the C-terminal domain of Cx43,underwent significant dephosphorylation when incubation of the cardiomyocytes with 1.0 ?mol/L aconitine.Conclusion Under certain concentration conditions,aconitine can induce significant dephosphorylation of Cx43 in the cardiomyocytes of rat.

Aim To study the effect of blocking adenosine A_1 receptors on BDNF protein expression and the endoplasmic reticulum(ER) content of the rat hippocampus.Methods The effect of DPCPX on BDNF protein expression in granular cells of the rat hippocampus was observed using immunohistochemical method,and the effect of DPCPX on ER in neurons of the rat hippocampus was observed with transmission electron microscope.Results DPCPX(0.1 mg?kg~(-1),ip,15 d) significantly increased the number of BDNF positive cells in hippocampus granular cells.DPCPX(0.5 mg?kg~(-1),ip,15 d) significantly increased both the levels of BDNF protein and the number of BDNF positive cells in hippocampus granular cells(P

OBJECTIVE:To determine the concentration of dehydroisoandrosterone sulfate (DHEAS) in human plasma by LC-MS.METHODS:With estrogen sulfate (ES) served as an internal standard,the plasma samples were deproteinized with acetonitrile,extracted by solid phase,hydrolyzed and derivatized.Then the concentration of DHEAS was determined by HPLC-MSD on Agilent SB C18 with column temperature kept at 40℃.The mobile phase consisted of acetonitrile-water (in gradient elution).Atmosphere pressure chemical ion source in negative ion detection model was employed.The ions selected for SIM (selected ion monitoring) quantitative analysis included m/z 490.0 (DHEAS ) and m/z 472.1(ES)[M-H]-.RESULTS:The linear range of DHEAS was 250.0～320.0 ng?mL-1(r=0.999 4).The extraction recovery of the simulated human albumin samples ranged from 71.1%～78.9% and its relative recovery ranged from 98.3%～101.4%.Both the intra-day RSD and inter-day RSD were less than 10%.The mean concentration of DHEAS in 15 health aged male volunteers was (981.6?353.4) ng?mL-1.CONCLUSION:The method is simple,practical,accurate and sensitive,and it is applicable for the determination of plasma concentration of DHEAS.

Object To compare the content in corilagin of Phyllanthus urinaria L. harvested from ten different regions of China in different months. Methods The content of corilagin was determined by UV spectrophotometry, detective wavelength is 270 nm. Results The content of corilagin in P. urinaria from Hainan and Guizhou province was the lowest, 0 58%. The highest content was given by the plant from Xi'an zone, Shanxi province, 1 78%. The best harvest time is in August and September, content of corilagin is 1 24%. Conclusion The difference of the corilagin contents in P urinaria . harvested from different regions and in different harvest time in the same region is great.

OBJECTIVE: To evaluate the efficacy and safety of huperzine A for patients with mild and moderate Alzheimer disease(AD). METHODS: The randomized controlled trials (RCTs) comparing huperzine A with placebo retrieved from Medline, Embase, the Cochrane Library, CBMdisc and CNKI were enrolled. The methodology quality of the included studies was evaluated and a Meta-analysis was performed using RevMan4.2 software. RESULTS: A total of five RCTs were included. The Meta-analysis results showed that as compared with placebo, huperzine A treatment significantly increased the scores of MMSE, decreased the scores of ADL(activities of daily living), moreover, the associated therapeutic effect increased gradually with the prolonging of the treatment time yet with small publication bias; huperzine A treatment significantly increased the scores of MQ(memory quotient), but some publication bias might be existed because of the small number of RCTs included. The sensitivity analysis on the comparison of HDS score changes showed that the influence of the quality of the RCTs could reverse the results. The comparison on effective rate showed that huperzine A was more effective than placebo. The comparison on safety showed that huperzine A was more likely to induced side effects than placebo, but the publication bias was big because there was only a few literature included. CONCLUSION: According to the Meta-analysis’s results, we think huperzine A is effective in improving memory and cognitive function of AD patients, and there is significant difference in clinical efficacy as compared with placebo, and there is small publication bias. In terms of safety, huperzine A has higher incidence of side effects than placebo, but the side effects are mostly mild and have little impact on the treatment. The above conclusion may serve as reference for clinical medication.

Objective To investigate the protective effects of combination alprostadil and dexmedetomidine preconditioning treatment on skeletal muscle ischemia reperfusion and secondary lung injuries.Methods Sixty patients aged less than 70 years old undergoing unilateral lower extremity surgery were randomly assigned into four groups:control group(group A),dexmedetomidine group(group B),alprostadil group(group C)and combined treatment group(group D) with 15 cases in each.In group C and D,10 μg alprostadil was given from vein at 15 min before tourniquet inflation,in group B and D 1 μg/kg dexmedetomidine was given from vein at 10 min before tourniquet inflation,the equal volume of normal saline was infused in control group.The blood samples were drawn from vein and artery for blood gas analysises and determination of malondialdehyde(MDA) and human pulmonary surfactant specific protein D (SP D) concentrations at the time before oxygen inhalation(T1),2 h after tourniquet deflation(T2),6 h after tourniquet deflation(T3) respectively.Results In group B,C and D,the alveolar arterial oxygen pressure difference(PA-a DO2)and respiratory index(R1) were significantly lower than those in group A at T2(P＜0.05).PA-aDO2 and R1 were significantly higher at T3 than those at T1 in every groups(P＜0.05);In group B,C and D,MDA and SP D were significantly lower than those in group A at T2 and T3(P＜0.05).In group D,MDA and SP D were significantly lower than those in group B and group C at T3(P＜0.05).Conclusion Alprostadil and dexmedetomidine are infused from vein preconditioning can attenuate the damages of skeletal muscle ischemia reperfusion and secondary lung injuries.The combination of alprostadil with dexmcdetomidine can produce stronger effects.

OBJECTIVE:To optimize the formulation of Asiaticoside cationic liposomes,and to investigate the characteristics of drug release in vitro. METHODS:The thin film dispersion method was used to prepare liposome;using encapsulation efficiency and drug-loading amount as index,the formulation of Asiaticoside liposomes was optimized by central composite design-response surface method with the ratio of drug to lipid(X1),the ratio of cholesterol to lipid(X2)and the concentration of D-mannose(X3) as factors. Using sodium lauryl sulfate as medium,in vitro release characteristics of cationic liposomes prepared with 1%octadecyl-amine was investigated by bag filter method,and compared with those of Asiaticoside solution and common liposome. RESULTS:The optimal formulation was X1 0.07,X2 0.17 and X3 0.03 g/ml. The encapsulation efficiency was (75.529 ± 1.071)%,and drug-loading amount was(2.539±0.029)%(n=3);the deviation from the predicted values were -0.217% and 0.205%;1% oc-tadecylamine was add into formulation to obtain cationic liposomes,and the Zeta potential had changed from -5.6 mV to 20.8 mV. in vitro accumulative release rates of Asiaticoside solution,common liposomes and cationic liposomes were 89.13%(12 h), 87.58%(72 h) and 94.46%(72 h),and the latter was in line with Weibull model. CONCLUSIONS:Asiaticoside cationic lipo-somes have high encapsulation efficiency,and can releases for 72 h.

AIM: To explore the different inhibitory effect of arsenic trioxide (As_2O_3) on hepatocarcinoma cell growth in SMMC-7721 and BEL-7402 cell lines and its mechanism. METHODS: The cell culture and trypan blue staining were used to study the inhibitory effect of arsenic trioxide on cell growth, and the glutathione (GSH) contents in hepatocarcinoma cells treated with arsenic trioxide were detected. RESULTS: Arsenic trioxide inhibited the growth of BEL-7402 cells in a time and dose-dependent manner. The inhibitory effect was significant at a lower dose of 0.50 ?mol/L for 24 h, however, to SMMC-7721 cells, a higher dose of 2.00 ?mol/L for 96 h was needed. The inhibitory rate of arsenic trioxide (0.25-2.00 ?mol/L) on BEL-7402 cell growth was higher than that on SMMC-7721 cells. The content of GSH in SMMC-7721 cells was much higher than that in BEL-7402 cells [(50.8?5.2) (?mol/g) protein and (18.7?1.4) ?mol/g protein, respectively]. CONCLUSION: There was a significant difference in inhibition of hepatocarcinoma cell growth by arsenic trioxide between BEL-7402 and SMMC-7721 cell lines, the cause of which may be due to the difference in GSH content in BEL-7402 and SMMC-7721 cells. [

Objective:To examine the content changes of neochlorogenic acid, chlorogenic acid, 1, 5- dicaffeoylquinic acid and total phenolic acids in the water extract from raw and fried Xanthii Fructus. Methods:The stir-frying method was used to process fried Xanthii Fructus from different habitats. The contents of neochlorogenic acid, chlorogenic acid and 1,5-dicaffeoylquinic acid in the wa-ter extract were determined by HPLC, the total phenolic acids content was determined by UV. Results:The contents of neochlorogenic acid, chlorogenic acid, 1,5-dicaffeoylquinic acid and total phenolic acids in the water extract from fried Xanthii Fructus were all in-creased. Conclusion:Fried Xanthii Fructus can increase the contents of effective ingredients in the decoction resulting in the enhanc-ment of clinical curative effect.