To evaluate the immune response and safety of the therapeutic DNA vaccine against the hepatitis B virus in rhesus macaques immunized by intramuscular injection of therapeutic DNA vaccine combined with electroporation. HBV DNA vaccine was administered to rhesus macaques by repeated intramuscular injections combined with electroporation in doses of 0 2,1 and 2mg, monthly for three consecutive months. The anti HBs antibody level was used to evaluate the efficacy of induction of humoral immunity. After that, the vaccine was intramuscularly injected daily in the same dose for six days to evaluate its safety. It was found that immunization with the therapeutic DNA vaccine against hepatitis B virus by means of intramuscular injections combined with electroporation induced a strong and specific anti HBs humoral immune response in rhesus macaques.Nine repeated intramuscular injections of the vaccine did not show adverse effects on behaviors, hematology, blood chemistry, and histopathology. No evidences of autoimmune mediated pathology and anti nuclear antibodies were observed in the rhesus macaques. These results suggested that electroporation significantly improves the efficacy of the therapeutic HBV DNA vaccine in rhesus macaques.The vaccine is safe and well tolerated.

OBJECTIVE:To preparation Ru'an mixture,and establish its quality standard and observe its therapeutic efficacy.METHODS:Ru'an mixture was prepared with Radix Bupleuri and Ramulus Cinnamomi and Radix Angelicae Sinensis as raw material.Radix Bupleuri,Radix Paeoniae Alba,and Radix Angelicae Sinensis were identified by TLC and the content of Tanshinol was determined by HPLC.RESULTS:The TLC spots of Radix Bupleuri,Radix Paeoniae Alba,and Radix Angelicae Sinensis were all clear.The linear range of Tanshinol was 0.203～2.030?g(r=0.999 8).The total effect rate in Ru' CONCLUSION:Ru'an mixture is reasonable in preparation technique,controllable in quality.

Objective To construct a RhoA-siRNA expression vector and determine its role on the malig-nant behavior of HepG2 cells.Methods A RhoA-siRNA DNA fragment was synthesized and cloned into the expression vector of pGenesil-1.The constructed Rhon-siRNA DNA plasmid was stably transfected into HerG2 cells by lipofectamine,and then HepG2 cells were divided into the HepG2/RhoA-siRNA group (HepG2 cells were transfected with pGenesil-1-RhoA-siRNA),HepG2/control group(HepG2 cells were transfected with control plasmid) and HepG2 group (without plasmid transfection).The inbibitory effect of RhoA-siRNA on RhoA protein expression was shown by Western blot.The proliferation,migration,growth potentiality and cell cycle of transfected HepG2 cells were evaluated by MTT assay,wounded healing,the plate cloning formation test and flow cytometry,respectively.All data were analyzed by one-way analysis of variance (ANOVA) and chi-square test.Results The expression of RhoA protein in the HepG2/RhoA-siRNA group was,significantly decreased compared with that in the other two groups (F=178.19,P<0.05).Scratched cells were healed within 48 hours in the HepG2/control group and HepG2 group,but not in the HepG2/RhoA-siRNA group.The clone formation rates in the HepG2/RhoA-siRNA group,HepG2 group and HepG2/control group were 39%±3%,67%±5%and 70%±6%,respectively,with a significant difference among the three groups(χ2=33.34,38.69,P<0.05).Flow cytometry showed that the number of cells transfected with RhoA-siRNA was highest in the G0/G1 phase and lowest in the S phase(F=70.46,76.57.P<0.05).Conclusion The RhoA-siRNA expression vector can effectively suppress the proliferation and migration of HepG2 cells,which may provide a novel gene therapy for hepatocellular carcinoma.

Objective To investigate the prevalence of healthcare-associated infection (HAI)in a hospital in 2013-2015,and provide reference for the improvement of HAI management.Methods Prevalence rates of HAI among patients in the First Affiliated Hospital of Zhengzhou University in 2013 -2015 were investigated with cross-sec-tional survey method,prevalence rates of HAI and constituent ratios of HAI sites in patients of different genders and different age groups were analyzed.Results In 2013 -2015,29 605 patients should be investigated,29 581 (99.92%)were actually investigated.Prevalence rates of HAI in 2013-2015 were 2.83%,2.14%,and 1 .73%,re-spectively,difference was significant(χ2 =27.521 ,P <0.01),which showed a downward trend year by year.Preva-lence rates of HAI in male was significantly higher than that of female (2.47% vs 1 .98%,χ2 =7.954,P <0.01). The prevalence rates of HAI in the age dimension was roughly U-shaped distribution in 2013 -2015.Prevalence rates of HAI in ≥70 age group was the highest(4.15%),in 0 -30 age group decreased with age,in > 30 age group increased with age.The top three sites for prevalence of HAI were lower respiratory tract (45.67%),upper respiratory tract (9.92%),and urinary tract (8.52%).Conclusion The prevalence rates of HAI decreased year by year in 2013-2015,which suggests that HAI prevention and control measures are effectively implemented.

OBJECTIVE To investigate the distribution of pathogenic bacteria in blood culture and their drug-sensitivity in our hospital in the past year.METHODS The blood samples were cultivated in the BacT/Alert 3D system and the isolated colonies were identified by the VITEK32 system or traditional routine procedures.The K-B method was carried out for the drug-sensitivity festing.RESULTS From the 716 blood samples received,we identified 109 strains of pathogens in which Gram-positive cocci accounted for 50.46%,Gram-negative bacilli for 44.04% and fungi for 5.5%.CONCLUSIONS There are many kinds of pathogens identified from blood culture and they have high resistance to many antibiotices.It is very important for doctors to get the results of blood culture in time in order to give the patients correct antibiotic therapy and to improve the cure rate and to control cross-infection in hospials.

Objective:Aimed to investigate the value of adrenocorticotropic hormone (ACTH) stimulation in adrenal venous blood sampling (AVS).Methods:Patients who diagnosed as primary aldosteronism (PA) and completed successful bilateral cannulation judged by selection index (SI) for routine and(or) ACTH stimulation AVS were enrolled. The lateralization index(LI) was calculated to compare the effect of ACTH stimulation on AVS cannulation success rate and lateralization judgment.Results:A total of 73 patients with PA were enrolled in the study, of whom 28 were confirmed as aldosterone producing adenoma (APA) after unilateral adrenalectomy. Cortisol and aldosterone in peripheral and adrenal veins were significantly increased after ACTH stimulation. The left SI was increased from 6.5(3.0-13.6) to 26.8 (16.9-40.3) ( P<0.01) and the right SI from 20.8(4.8-34.8) to 57.6(35.7-80.9) ( P<0.01) after ACTH stimulation. There was no significant difference on LI before and after ACTH stimulation [7.7(2.3-19.6) vs 5.6(1.9-14.6), P=0.14]. The success rates of left and right adrenal cannulation were increased by 15% and 10% respectively after ACTH stimulation. For 57 patients who were determined in successful cannulation by both routine and ACTH stimulation AVS, 27 patients were determined to have lateralization by both AVS methods, 21 patients were determined to have bilateralization, and the consistency of lateralization by both AVS methods was 84%(48/57). Among the 28 patients who were confirmed to be APA after unilateral adrenalectomy, the correct rate of lateralization by both AVS methods was 89% (25/28). Conclusion:ACTH stimulation is able to improve the success rate of bilateral adrenal vein cannulation, and is helpful to judge AVS results. For patients with successful cannulation, there is no significant difference in lateralization judgment for routine and ACTH stimulation AVS.

To investigate the renal venous and spermatic venous sampling in assisting the diagnosis of reninoma. The case of reninoma was retrospectively reviewed together with the literature review of reninoma diagnosed with renal venous and spermatic venous sampling. A young patient with hypertension and headache was admitted to our hospital. Laboratory test showed high plasma renin concentration (>500 mIU/L), and enhanced computed tomography(CT) in the upper abdomen showed a mass in left inferior renal pole. The concentration of renin in the left spermatic vein was significantly higher than that in renal veins and branches, and peripheral vein, which was considered the left reninoma possibility. The left renal mass was resected surgically and pathologic exam revealed reninoma. The renin level and the blood pressure recoveried normal after operation.The literature reported that the positive rate of renal vein segmental blood collection for locating renin tumor was only 8.3%-64%. The possible reason was that reninoma usually located in the renal cortex, and the tumor blood might be collected by renal capsule vein instead of renal vein. In fact, the renal capsule vein intersects with the lateral division of the spermatic vein, but there have been no reports about the localization of reninoma by spermatic vein sampling. Since renin secreted by reninoma may go into the spermatic vein through renal capsule vein, it should be noted that spermatic venous blood should be collected simultaneously in renal vein sampling when locating reninoma.