0.05);the costs were 459.00 yuan,481.68 yuan and 789.48 yuan,respectively,showing significant differences between Group A and Group C or between Group B and Group C(P0.05).CONCLUSION: Group A is preferable in terms of cost-effectiveness,while there was no significant difference between Group A and Group B in cost.

Objective To construct DNA vaccine expressing HIV-1 AE2f gp145-tat-rev-nef fusion gene( AE-Gp145TRN) and to compare the immunogenicities of DNA vaccines expressing Tat, Rev and Nef in gene fusion formulations of tat-rev-integrase(c-half)-vif-nef( AE-TRIVN) and AE-Gpl45TRN. Methods DNA vaccine was constructed by inserting the codon optimized HIV-1 AE2( gp145-tat-rev-nef fusion gene into mammalian expression DNA vector. In vitro expression efficiency of the constructed DNA vaccine was determined by Western blot and the immunogenicities of AE-Gpl45TRN and AE-TRIVN were compared by immunizing female BALB/c mice. IFN-r ELISPOT assay was used to read out the specific T cell immunity. Results Western blot assay showed the constructed DNA vaccine could be expressed efficiently in vitro. After vaccination, AE-TRIVN mounted significantly higher T cell responses against Tat, Rev and Nef[(148±91)SFCs/106 splenocytes]than Gpl45TRN[(55±28) SFCs/106 splenocytes]. Specific T cell responses elicited by AE-TRIVN predominantly targeting Rev, whereas Gpl45TRN could significantly enhance T cell responses against Nef. Conclusion AE-TRIVN and Gpl45TRN induced distinct T cell response modalities, which implied different gene fusion formulations may affect the immunogenicity of specific DNA vaccines.

Objective To evaluate the effect of surgical trauma on the cognitive function and expression of hepcidin and ferroportin 1 (FP1) in hippocampus in aged rats.Methods One hundred male Sprague-Dawley rats,aged 18 months,weighing 400-500 g,were randomly divided into 2 groups with 50 rats in each group:control group (group C) and surgical trauma group (group ST).The rats were anesthetized with chloral hydrate,but underwent no operation in group C.The rats Were anesthetized with chloral hydrate and underwent 30 min of modified exploratory laparotomy in group ST.Ten rats were chosen from each group at 24 h after operation and the cognitive function was assessed using Morris water-maze test for 6 consecutive days.Ten rats were sacrificed on 1st,3rd,5th and 7th days after beginning of Morris water-maze test and brains were removed for determination of hepcidin and FP1 expression in hippocampus by PCR and Western blot.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant and frequency of crossing the original platform were decreased on 3rd,4th and 5th days after beginning of Morris water-maze test,and the expression of hepcidin was up-regulated and the expression of FP1 was down-regulated at each time point in group ST (P ＜ 0.05).Conclusion Surgical trauma can decrease the cognitive function in aged rats and the mechanism may be related to up-regulation of hepcidin expression and down-regulation of FP1 expression in hippocampus.

Objective To investigate the effect of surgical trauma on the cognitive function and activation of microglias in hippocampus in rats of different ages.Methods Seventy-two male Sprague-Dawley rats,aged 3-4months,were randomly allocated into 2 groups:adult control group (n =30) and adult surgery group (n =42).Seventy-two male Sprague-Dawley rats,aged 18-20 months,were randomly allocated into 2 groups:aged control group (n =30) and aged surgery group (n =42).The rats were anesthetized with 5% chloral hydrate 4-6 ml/kg and underwent exploratory laparotomy in surgery groups,while normal saline 1 ml/kg was injected intraperitoneally in control groups.Morris water maze test was performed at 1-7 days after surgery.Fear conditioning test was performed 1 day after surgery to evaluate the space and fear memory abilities.The animals were sacrificed on 1st,3rd and 7th days after surgery and hippocampi were removed for measurement of OX42 expression in microglias by immunohistochemistry.Results Compared with adult control group,the percentage of freezing time in total time was significantly decreased,and OX42 expression in microglias was up-regulated on 1st day after surgery (P ＜ 0.05),and no significant change was found in the escape latency and the number of crossing the original platform in adult surgery group (P ＞ 0.05).Compared with aged control group,the escape latency was significantly prolonged,the number of crossing the original platform was decreased,the percentage of freezing time in total time was decreased,and OX42 expression in microglias was up-regulated on 1st and 3rd days after surgery in aged surgery group (P ＜0.05).Conclusion Surgical trauma decreases fear memory ability,but exerts no effect on the space memory ability in adult rats.Surgical trauma decreases the space and fear memory abilities in aged rats,which maybe related to activation of microglias in hippocampus.

Objective To investigate the clinical practice pressure and mental health of medical students with type D personality.Methods Type D Scale-14 (DS14) and Beck-Srivastava Stress Inventory (BSSI) test were applied to 371 medical students to assess the personality types and pressure.The symptom checklist 90 (SCL-90) was used to evaluate the psychological health.Results ①The detection rate of type D personality of medical students was 36.39％.②The average score in BSSI of medical students of type D personality was (99.27± 10.51),which was higher than medical students of non-type D personality (87.60± 11.37),and the difference was statistically significant (t=9.9711,P=0.0000).The medical students' score of type D personality in SCL-90 of 9 factors were all higher than medical students of non-type D personality,but the statistically significant difference were only in the score of depression,anxiety and psychosis-like symptoms (t=2.4409,P=0.0151;t=2.8662,P=0.0044;t=2.7783,P=0.0057).Conclusion In face of the same pressure of medical clinical practice,the medical students of type D personality are more likely to have a heavier psychological burden,and the college should pay special attention to the problem and try to intervene the problem,so as to reduce the pressure caused by a variety of psychological problems.

Objective To investigate the effect of dexamethasone on the postoperative cognitive function in rats.Methods One hundred and eighty Sprague-Dawley rats,aged 18-20 months,weighing 400-600 g,were randomly allocated into 3 groups (n =60 each)∶ control group (C group),surgery group (S group) and dexamethasone group (D group).In groups S and D,the rats were anesthetized with 5％ chloral hydrate 4-6 ml/kg and underwent abdominal surgery.The rats in group D received intraperitoneal injection of dexamethasone 10 mg/kg at the beginning of anesthesia,while the rats in group C underwent no surgery and received intraperitoneal injection of normal saline 1 ml/kg instead.Six rats in each group were chosen at 3 h and 7 days after surgery and sacrificed,and their brains were immediately removed for detection of the expression of OX42 (a specific marker for activation of microglia) in hippocampus.Another 6 rats in each group were chosen at 3 h,and 1,3 and 7 days after surgery and sacrificed,and their brains were immediately removed for detection of the expression of IL-1β mRNA and TNF-α mRNA in hippocampus.Cognitive function was assessed by Morris water maze test and fear conditioning test.Results Compared with group C,the escape latency was prolonged,the frequency of crossing the original platform was decreased,the postoperative freezing time was shortened,and the expression of OX42 after surgery and IL-1β mRNA and TNF-α mRNA at 3 h and 1 and 3 days after surgery was up-regulated in groups S and D (P ＜ 0.05 or 0.01).Compared with group S,the escape latency was shortened,the frequency of crossing the original platform was increased,the postoperative freezing time was prolonged,and the expression of OX42 at 3 h after surgery and IL-1β mRNA and TNF-α mRNA at 3 h and 1 and 3 days after surgery was down-regulated in group D (P ＜ 0.05 or 0.01).Conclusion Dexamethasone can inhibit the over-activation of microglia and reduce the inflammatory response,thus improving cognitive function in rats.

Objective To investigate the variation of fentanyl concentration and the gene expression and activity of rat intestinal cytochrome P450 3A1 in anhepatic phase. Methods The experiment was comprised of 2 steps. Step 1: The rats were randomly divided into experimental group (group A2, underwent occlusion of the hepatic portal) and control group (group A1), with 10 rats in each group. Fentanyl blood concentration was analyzed by LC/MS/MS. Step 2: The rats were randomly divided into group B1 (control), group B2 and group B3 (the rats underwent devascularization of the hepatic portal for 30 or 60 min). The levels of CYP3A1 in rat small intestine were assessed with RT-PCR and the enzymic activity of CYP3A1 was detected by fluorometry. Results Fentanyl concentration in anhepatic phase dropped more slowly in group A2 than group A1 (P

BACKGROUND: The connection between Natural killer(NK)-cells and allogeneic bone marrow transplantation(allo-BMT)has aroused increasing attention.OBJECTIVE: To explore the effect of NK cells on graft rejection,hematopoietic and immune reconstitution in mouse undergoing allo-BMT.METHODS: Lethally and nonlethally irradiated BALB/c(H-2d)mice were transplanted with C57BL/6(H-2b)bone marrow plus donor peripheral T cells and/or NK cells.RESULTS AND CONCLUSION: Compared with lethally irradiated and allo-BMT group without infusion of NK cells,the survival rate in lethally irradiated and allo-BMT group with infusion of NK cells significantly enhanced; leukocytes count,expression level of CD19+and CD34+cell count recovered rapidly; expression level of H-2b*cell obviously increased.Expression level of CD34"cell in the group with infusion of NK cells was obviously lower than that of the group without infusion of NK cells at 28 days after transplantation,but there was no significant difference between the 2 groups at 60 days(P > 0.05).In nonlethally irradiated and allo-BMT group without NK cell infusion,expression level of H-2b*cell significantly decreased at 30 days after transplantation,and reduced to before transplantation level at 60 days; while expression of H-2b+cell yet could be detected with more than 80% at 60 days after transplantation in group infused with high and low concentration of NK cells.In alIo-BMT mice,alloreactive NK cell inhibits graft rejection,enhances engraftment,promotes the reconstitution of hematopoiesis and immunity,and increases survival rates.

Objective To analyze CD127 expression on the memory CD8+ lymphocytes from hepatitis B e antigen (HBeAg) positive chronic hepatitis B (CHB) patients treated with peginterferon α-2a (Pegasys). Methods Thirty HBeAg positive CHB patients were treated with peginterferon α-2a 180 μg once a week for 48 weeks and followed up for 24 weeks. The memory CD8+ lymphocytes were characterized by expressing CD45RA and CD27 markers. CD127 expression on cell surface was measured by four-colour flow cytometry. The difference of mean values between groups was evaluated by Mann-Whitney test. Results The CD127 expression on CD8+ T lymphocytes was significantly lower in HBeAg positive CHB patients compared to healthy controls (Z=2.889, P＜0.05), which was negatively correlated with serum hepatitis B virus (HBV) DNA level and HBeAg titers. The CD127 expression increased along with the decrease of HBV DNA and HBeAg after 24-week, 48-week and 72-week treatment in patients showing good response to peginterferon α-2a, while CD127 expression didn't change markedly in non responders (Z24w = 1.954, Z48w = 2.789, Z72w = 2. 989; all P＜0. 05). Conclusion CD127 expression on memory CD8+ lymphocytes increases along with effective anti-HBV treatment in CHB patients, which can be used as a marker for evaluating the effectiveness of anti-viral treatment.

Objective To compare the 2-year efficacy of de novo combination therapy with lamivudine (LAM) and adefovir dipivoxil (ADV) to that of entecavir (ETV) monotherapy in treatment of patients with hepatitis B virus ( HBV )-related decompensated cirrhosis.Methods A total of 120 naive patients with HBV-related decompensated cirrhosis admitted to Shangyu People's Hospital and the First Affiliated Hospital of Zhejiang University from January 2007 to April 2008 were enrolled,in which 60 were treated with LAM and ADV combination therapy,and other 60 patients were treated with ETV monotherapy.Tests for liver and kidney function,alpha-fetoprotein,HBV serum markers,HBV DNA load,prothrombin time (PT),and ultrasonography or CT scan of liver were performed every 1-3 months.Repeated measure ANOVA and x2test were used to compare the efficacy,side effects and accumulated survival rates at 12 and 24 month in two groups.Results Forty-five patients in each group were followed-up for 24 months.There was no significant difference in HBV DNA negative rates and ALT normalization rates at month 12 (x2 =2.12 and 2.88,P ＞0.05 ) and month 24 between two groups (x2 =3.21 and 3.24,P ＞ 0.05); while HBeAg seroconversion rate in LAM + ADV group at month 24 was significantly higher than that in ETV group (43.5％ vs.36.4％,x2 =4.09,P＜0.05).Viral breakthrough occurred in 2 cases (4.4％) by month 12 and 3 cases (6.7％) by month 24 in LAM + ADV group,and no viral mutation was observed; while in ETV group,viral breakthrough occurred in 1 case ( 2.2％ ) by month 12 and 2 cases (4.4％) by month 24,and viral mutation was observed in 1 case (2.2％) by month 24.At the end of month 24,increase of AIb (F=18.9 and 17.3,P＜0.05),decrease of TBil and ALT (F=16.5,17.1 and 23.7,24.8,P ＜0.05 ),shortening of PT ( F =22.7 and 24.5,P ＜ 0.05 ),and the improvements of CTP and MELD scores (F=18.5,17.8 and 24.2,23.8,P＜0.05) were observed in both groups.The accumulative rates of mortality or liver transplantation were 16.7％ ( 10/60 ) and 18.3％ ( 11/60 ) in LAM + ADV and ETV groups,respectively.No blood creatinine increased above the normal upper limit was observed in both groups.Conclusion Both LAM + ADV combination therapy and ETV monotherapy can effectively inhibit HBV replication,improve liver function,decrease mortality and viral resistance,but the 24-month HBeAg seroconversion rate in combination therapy group is higher than that in monotherapy group.