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.

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.

Analogy is to construct the theory of traditional Chinese medicine in clinical thinking method,which the proper use of this thinking is helpful to improve the clinical thinking of Chinese medicine,contributes to the development of disease differentiation and treatment ideas.This article tried to analyze the diagnosis and treatment of kidney diseases with Professor Zhu Zongyuan on the process,to explore the analogy on disease differentiation treatment guide.

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.

BACKGROUND: Severe acute respiratory syndrome (SARS) is a kind of very strong infective respiratory system diseases. Due to rapid development of disease course, severe symptoms, forced isolation, being away the supports from relatives and friends, many patients with SARS presented extremely anxiety and panic, even depression and despair, and they are in the crisis state and need mental support and help.OBJECTIVE: To evaluate the therapeutic effect of mental crisis intervention in patients with SARS.DESIGN: Controlled observation.SETTING: Xiaotangshan Hospital of Chinese PLA.PARTICIPANTS: Totally 680 patients with SARS who received the mental intervention in the Psychological Counseling Center, Xiaotangshan Hospital of Chinese PLA in May 2003. The patients, 379 male (55.74%)and 301 female (44.26%), were aged (35±7) years. Degree of education:34.5% of patients received education below junior high school, 31.5% senior high school and 34.0% college. 100% of the patients had family member or colleague-infected life events, and 14.4% of the patients lost their family members. Degree of disease: 6.9% of patients were in the severe period, 67.5% in the stable period and 25.6% in the rehabilitation period.METHODS: ① Totally 489 patients who received telephone counseling,and attended telephone mutual-help group, group debugging, individual counseling and group counseling were chosen, serving as intervention group, and the other 191 patients served as control group. ②The mental health state of patients in the 2 groups was evaluated at admission with symptom checklist 90 (SCL-90), which consisted of somatization, obsessive compulsive, interpersonal sensitivity, depression, anxiety, hostility, phobic anxiety, paranoid ideation and psychoticism 9 factors. The grading scoring was used. The higher grades, the more severe symptoms. ③ The patients of intervention group were given comprehensive mental intervention, which included handing out materials(self-debugging propagandist manual), care from staff, mental counseling by telephone, group debugging, individual counseling, guidance by mental doctor, etc. Patients in the control group only received handed materials (self-debugging propagandist manual) and the care from staff. SCL-90 was used to re-evaluate the patients of intervention group and control group before discharge. ④Intervention therapeutic effect of patients in 2 groups were evaluated with comprehensive mental intervention questionnaires, which were mainly about self and social cognition, feeling and emotion, life behavior styles, physiological and body symptoms 4 dimensions. Grading scoring was used. The higher points, the better intervention effect. Self-rating and rating by others were used in each question. If the results in self-rating are the same as those in rating by others, questionnaire is valid, otherwise, it is invalid). ⑤ The scoring of all the patients at admission was compared with that of Beijing norm by using SCL-90.The mental health of patients of 2 groups after intervention was measured, evaluated and compared with SCL-90.MAIN OUTCOME MEASURES: Difference in scoring of SCL-90 between all the patients and Beijing norm; Changes in the scoring of SCL-90 between intervention group and control group at admission and after intervention.RESULTS: Totally 680 comprehensive mental intervention questionnaires were handed out, and 669 of them were retrieved, among which 483 from intervention group and 186 from control group. Altogether 669 patients participated in the result analysis. ① Points of each factor in SCL-90 of patients at admission were significantly higher than norm, especially in anxiety and phobic anxiety. 45.5% and 50.1% of the investigated patients had more than 3 points in anxiety and phobic anxiety respectively. 86.5% of the investigated patients had mental problems to different degrees. ②There was no significant difference in the points of various factors in SCL-90 at admission between intervention group and control group (P＞0.05).In the various factors of SCL-90, no significant difference in the points of various factors in SCL-90 at admission between intervention group and control group (P＞0.05).In the various factors of SCL-90,no significant difference exists in phobic anxiety and paranoid ideation and psychoticism between two groups (P＞0.05),but the points of the other factors of intervention group were significantly lower than those of control group (P＜0.05-0.01).CONCLUSION: Mental crisis intervention can remarkably lighten the somatization, obsessive-compulsive, anxiety and depression, lower interpersonal sensitivity and improve hostility.

Objective To explore the effect of aluminium on the concentration of intracellular Ca2+ in the hippocampus neurons of postnatal rat. Methods The hippocampus neurons of postnatal rats were primarily cultured and treated with Al3+ at doses of 0,10,100 and 1 000 ?mol/L. Fura-2-AM calcium ions fluorescence indicator was used to determine the concentration of the intracellular Ca2+ when they were cultured for 15 min,30 min and 45 min. Results The concentration of intracellular Ca2+ in the hippocampus neurons increased with the aluminum concentration increased and as the concentration of the aluminium was 1 000 ?mol/L,at the three cultured time points,the concentration of intracellular Ca2+ were (276.2?53.4) nmol/L,(306.5?73.7) nmol/L and (408.8?79.7) nmol/L respectively,significantly higher than the control group(P