An optimized recombinant HPV16 E6E7 fusion gene(HPV16 ofE6E7)was constructed according to codon usage for mammalian cell expression,and a mutant of HPV16 ofE6E7 fusion gene(HPV16 omfE6E7)was generated by site-directed mutagenesis at L57G,C113R for the E6 protein and C24G,E26G for the E7 protein for HPV16 ofE6E7 [patent pending(CN 101100672)].The HPV16 omfE6E7 gene constructed in this work not only lost the transformation capability to NIH 3T3 cells and tumorigenicity in SCID mice,but also maintained very good stability and antigenicity.These results suggests that the HPV16 omfE6E7 gene should undergo further study for application as a safe antigen-specific therapeutic vaccine for HPV16-associated tumors.

Objective: To investigate the effect of interleukin-8 (IL-8) on survival and apoptosis of hypoxic human umbilical vein endothelial cells (HUVECs) with its mechanisms.
Methods: Hypoxic HUVECs were induced by CoCl2. HUVECs were cultured with different concentrations of CoCl2 or IL-8 and the survival rates of HUVECs were examined. Normal or hypoxic HUVECs were incubated with IL-8 or simultaneously incubated with anti-IL-8 or Akt inhibitor LY294002 for 48h. The HUVECs survival rate was detected by MTT method, apoptosis rate was measured by Annexin V-FITC/PI method, the protein expressions of Caspase-3, phosphorylated Akt (pAkt) and GSK-3βser9 (pGSK-3βser9) were evaluated by Western blot analysis.
Results: By 12 h incubation with low concentration of CoCl2 (50, 100 μmol/L), HUVECs proliferation were improved, P<0.01; while by 24h and 48 h incubation with high concentration of CoCl2 (200, 400 μmol/L) HUVECs survival were decreased. By 48h and 72h incubation with IL-8 at (10, 50, 100 ng/ml), the hypoxia induced cell apoptosis was decreased and the survival was increased,P<0.01. IL-8 at 10 ng/ml may obviously inhibit the normal and hypoxic HUVECs apoptosis, down-regulate caspase-3 and up-regulate pAkt and pGSK-3βser9 expressions in HUVECs,P<0.01, while the above effects could be reversed by LY294002 and anti-IL-8,P<0.01.
Conclusion: IL-8 could down-regulate caspase-3 and up-regulate pAkt and pGSK-3βser9 expressions in HUVECs and therefore, inhibit the apoptosis and improve the survival of hypoxic HUVECs.

Objective To investigate the infection following the lymphocytes deleted agent (ATG) and IL-2 receptor antagonists (Basilixinab and Daclizumab)-based induction therapy after renal trausplantation.Methods A retrospective analysis was carried out on 701 kidney transplant recipients between Jan. 1,2005 to Dec.31,2010.According to exclusive and inclusive criteria,finally 549 patients were evaluated,including 429 patients treated with ATG (ATG group) and 120 patients with anti-CD25 monoclonal antibodies (monoclonal antibodies group; 86 patients with Basiliximab,and 34 patients with Daclizumab).The incidence of acute rejection,infection rate,infection time,hospital stay,severe infection rate and mortality were analyzed.After operation,the patients received an immunosuppression therapy including Tacrolimus (cyclosporine A),Mycophenolate-Mofetil and prednisone to present rejection. Part of the patients were treated with ganciclovir and sulfamethoxazole sulfadiazine and trimethoprim for infection prevention.Results The acute rejection rate in ATG group and monoclonal antibodies group was 15.9％ (68/429) and 10.0％ (12/120),and there was no statistically significant difference (P＞0.05).The infection rate in ATG group was 11.9％ (51/429),including 13.7％ (7/51) with severe infection,and mortality was 7.8％(4/51).The infection rate was 15.0％ (18/120) in monoclonal antibodies group,including 11.1％ (2/18) with severe infection,and mortality was 5.6％ (1/18).There was no statistically significnat difference in infection rate,severe infection rate and mortality between two groups (P＞0.05).The hospital stay in ATG group and monoclonal antibodies group was 25.8 days and 19.1 days respectively (P＜0.05).Dead cases had not received regular anti-infection treatment,and the patients age was over 50 years.Conclusion The infection risk and mortality between these two induction therapies are identical,but hn comparison to the patients using ATG,the infection of patients using anti-CD25 monoclonal antibodies is easier to control.Anti-infection prophylaxis is important to reduce infection rate and decrease infectious mortality.

objective To evaluate the influence of HLA matching and new immunosuppressants on pre-venting acute rejection of renal allograft in sensitized recipients. Methods 751 recipients underwent renal transplantation were enrolled in this study including 46 sensitized recipients (study group) with PRA be-tween 10%-90% and 705 non-sensitized recipients (control group) with PRA less than 10% pretransplant. All patients in the study group received induction course (ATG 100 mg/d, 5-7 d) plus triple-immunosup-pressive therapy including FK506 + MMF + steroid. The rate of acute rejection and delayed graft function after renal transplantation was analyzed. The influence of HLA matching on preventing acute rejection was al-so evaluated. Results The acute rejection rate in the study group and control group was 30.43% and 19. 57%, respectively, (P < 0.05). The rate of delayed graft function was 60.86% in the study group, signifi-cantly higher than that of the control group (11.87%). There was no statistically difference of one-year pa-tient / graft survival rotes between the two groups. The average serum creatinin levels at one-year posttrans-plantation were similar between the two groups (130 mmol/dl in the study group and 125 mmol/di in the control group). The average loci of HLA matching in the study group (4.2) was significantly higher than that in the control group (2.8). The acute rejection rate in the study group was significantly higher when lo-ci of HLA mismatch ranging from 2-4 compared with loci of HLA mismatch less than 2. The acute rejection rate was significantly higher in the highly sensitized recipients (PRA ranging from 50% -90% pretmnsplant) than that in the less sensitized (PRA ranging from 10% to 20% pretransplant) in the study group. Patients with higher PRA level posttransplantation were prone to developing acute rejection. Conclusion HLA matching and new immunosuppressants can reduce the incidence of acute rejection in sensitized recipi-ents and increase the survival rate of patients and allografts.

Objective To investigate the incidence of infection and the effect of anti-infection prophylaxis in renal transplanted patients after Basiliximab induction therapy. Methods A total of 204patients who have received renal transplantation and Basiliximab induction therapy from January 1,2001 to December 31, 2010 in our hospital have been retrospective analysed in this study. These patients were divided into a prophylaxis group (118 cases) with Ganciclovir + Sulfadiazine +Trimethoprim therapy and a control group (86 cases) without any anti-infection prophylaxis.Furthermore, 440 transplanted patients in the same peroid without any induction therapy were also analysed. They were also devided into two groups: an anti-infection prophylaxis group (206 cases)and a control group (234 cases) without any anti-infection prophylaxis. Results In the prophylaxis group with Basiliximab induction therapy, there were 23 patients (19. 5 %, 23/118)experienced hospitalization due to infection, 3 cases (13. 0 %,3/23) among them were severe infection, and 3patients (13.0 %, 3/23) died from vital infection. In the non-prophylaxis control group with Basiliximab induction therapy, 27 patients (31.4 %, 27/86) had infection complication, 7 patients (25.9 % ,7/27) among them were severe infection, and 4 patients(14. 8 % ,4/27)died. The incidence of infection between the above two groups is significantly different (P＜0. 05). In the prophylaxis group without induction therapy, the incidence of infection was 15.0 % (31/206), there were no severe infection cases but 7 patients (22. 6 %, 7/31) died from infection. In the non-prophylaxis control group without induction therapy, the incidence of infection was 12. 8 % (30/234), 3 cases among them were severe infection(10. 0 %,3/30)and 5 patients died from infection (16. 7 %, 5/30).The incidence of infection in Basiliximab induced patients without anti-infection prophylaxis is significantly higher than that in patients without induction therapy and anti-infection prophylaxis (31.4 % vs. 12.8 %,P＜0.01). Conclusion Basiliximab induction therapy increased the risk of infection, but not the rate of mortality. It is necessary to give anti-infection prophylaxis in renal transplanted patients with Basiliximab induction therapy.

Objective To evaluate the postoperative quality of life of donors in living donor renal transplantation patients.Methods One hundred and seventeen donors were involved in present study from 2006-2008.A crosssectional survey was performed with questionnaire research to all the donors who received living donor nephrectomy during this period.The questionnaire included sociodemographic characteristics,surgical complications,economic status,donors awareness status,family support,the health care,social welfare and daily exercise after surgery.The Chinese version of SF-36 was used as the measurement of quality of life.The statistic analyze include T test,analysis of variance and stepwise regression analysis.Results The donors' mental health status was better than the healthy population (P ＜ 0.05).The difference of quality of life and scores of other dimensions compared with the healthy population was not statistically significant (P ＞ 0.05).In univariate analysis,four kinds of fields such as age,education level,economic status and physical exercise were associated with quality of life.In further multivariate analysis,with exclusion of the interaction between various factors,the main factors for postoperative quality of life are the cultural,economic status and physical activity (P ＜ 0.05).Conclusions Social and psychological factor should be concerned in donor's preoperative screening.Good social psychological background,the necessary psychological intervention and postoperative follow-up maybe play an important role to improve the postoperative quality of life in living donor renal transplantation.

Objective To improve the technology of retroperitoneal laparoscopic living donor nephrectomy and observe its clinical effect.Methods Forty-one cases of living donors subject to nephrectomy by the new retroperitoneal laparoscopic technique from July 2009 to June 2012 were retrospectively.The new technique was modified as follows: (1) Alternate use of blunt dissection,sharp dissection and harmonic scalpel; (2) After separation of renal vein,artery and ureter,a 5-6 cm incision parallel to rectus abdominis from Trocar was made in order to put a hand inside retroperitoneum; (3) A biopsy of the kidney was made from Trocar with the help of a hand for holding the kidney; (4) Pulling the kidney with a proper strength and blocking renal artery and renal vein with Hem-o-lock,then cutting off them and taking out the kidney.Results Forty-one cases of live donors subject to nephrectomy were operated on successfully,and were not converted to open operation.The operative time was 65-130 min (mean 85 min).The warm ischemia time was 58-110 s (average 78 s).Living donor kidney artery length was 2.1-3.7 cm (average 2.9 cm).Living donor kidney vein length was 2.5-4.1 cm (average 3.5 cm).Blood loss was 15-80 ml (average 28 ml).Hospital stay after surgery was 4-7 days (average 4.8 days).All biopsy specimens were achieved from 41 cases.None suffered from complications except two cases of perilymphorrhea.Forty-one recipients recovered well after renal transplantation.Conclusion The improved retroperitoneal laparoscopic living donor nephrectomy is considered to be safe,effective and feasible.It is a good way to protect renal function and reduce injury.

Objective To analyze the dynamic changes of dendritic ceils (DCs) and their subsets plasmacytoid DC (pDC) and myeliod DC (mDC) in peripheral blood of renal transplantation patients,and to confer the relationship between DCs subsets and graft rejection.Methods White blood cells (WBC) and mononuclear cells (PBMNCs) in peripheral blood of 28 renal transplantation recipients (test group) were measured before operation and at 1st,7th,28th day after operation.The number of DCs and subsets,and pDC/mDC were detected by using flow cytometry,and IL-10 and IL-12 levels were determined by using ELISA before and after operation.Ten volunteers (control group) served as controls.Results The levels of DCs,pDC and mDC before operation in test group were lower than in control group (P＜0.05),but there was no statistically significant difference in pDC/mDC ratio between two groups (P＞0.05).The number of DCs in test group was significantly decreased on the first day after operation up to the lowest level,then slowly increased,and recovered 73.7 ％ at 28th day after operation.The number of mDC and pDC was also decreased after operation,but mDC recovered faster than pDC (P＜0.05).On the day 7th after operation,the number of mDC in the recipients with graft rejection was higher than in those without graft rejection in test group (P＜0.01 ).There was no significant difference before and after operation in the levels of IL-10 and IL-12 in test group.Conclusion The number of DCs and subsets are related to the recipients' immune state,and their abnormality displays unstable immune state of recipients.The number of DCs and subsets can be used as an assistance index to diagnose graft rejection.

Abstract Comprehensive sexuality education is an important part of quality education, primary and secondary schools are the most suitable places for sex education. This paper sorts out the current status of sexuality education for primary and secondary school students in developed countries after presenting the overall significance of school based sexuality education, and further points out the problems and urgency of sexuality education for primary and secondary school students in China. It also put forward the way to new directions for advocacy, including the comphrehensive sexuality education curriculum system, training of sexuality education teachers, the positive and active role of families, as well as social and community support for sexuality education in schools.

Objective To reveal the change trend of malignant tumor after renal transplantation in China based on the epidemiological and clinical features that were publicly reported in China in recent 10 years. Methods The search terms ‘renal transplantation’and ‘tumor’were searched on China Academic Journal Network Publishing Database and China Science Periodical Database to screen out the qualified researches strictly.General conditions,tumor sites and regional differences of malignant tumors were analyzed.Results Fifteen thousand one hundred and twenty cases from nine literatures published from 2003 to 2014 and a single-center experience of renal transplantation in Beijing Friendship Hospital of Capital Medical University were screened out.Four hundred and fourty-six cases had malignant tumor after renal transplantation with the total tumor incidence of 2.95% (446 /15 120) and the tumors were mainly urinary system tumors after transplantation (55.8%).Conclusions The total incidence of malignant tumor in renal transplant recipients is 2.95% and the urinary system tumors are most common.Thus,tumor screening after renal transplantation should be taken as the routine examination during follow-up.