ObjectiveTo construct one recombinant adenovirus AdE7E6 expressing HPV16 E6 and E7 fusion protein as candidate for HPV16 therapeutic vaccine.MethodsThe codon-optimized E6 and E7 gene,were fused to create one open reading frame,then inserted into adenovirus vector pCD316.A strain of recombinant adenovirus was constructed through homologous recombinant in 293 cells,and identified by PCR and Western blot.Finally,it was employed to study it's immunogenicity and the activity of the tumor growth regression.ResultsThe PCR result showed that E6E7 fusion gene had been integrated in recombinant Ad5 DNA.Western blot test confirmed that the E6E7 fusion protein was highly expressed in 293 cells infected with Ad5E7E6 recombinant adenovirus.The recombinant adenovirus elicited significant E7 specific CD8+ T lymphocyte response in vaccinated mice.These responses could completely prevent the TG-1 tumor cell bearing mice treated with AdE7E6 from developing into tumor.ConclusionThese results suggested that rAd5E7E6 could be a potent vaccine candidate for the treatment of HPV16-associated tumors and their precancerous transformations.

Objective To explore the value of medical three-dimensional visualization technology in preoperative precise assessment for hepatocellular carcinoma (HCC).Methods The clinical data of 24 HCC patients were analyzed retrospectively from Jan 2015 to Dec 2016 in our department.Thin-layer 320-slice CT scaning was performed and data stored in the form of DICOM.Three dimensional reconstruction and simulated hepatectomy were conducted,and correlation test analysis was performed.Results The volume of simulated resected liver was (565 ± 296)ml,while the actual liver resection volume was(523 ± 252)ml.There was no significant difference between the predicted and the actual values (t =1.734,P ＞ 0.05).The actual liver resection volume was positively correlated with the predicted liver resection volume (r =0.920,P ＜ 0.05).The stimulated resection margin was (1.2 ± 0.6)cm,while the actual resected tumor margin was (1.0 ± 0.5) cm.There was no significant difference between the predicted and the actual values (t =1.853,P ＞0.05).The actual margin was positively correlated with the predicted one (r =0.956,P ＜ 0.05).Conclusion The three-dimensional visualization technology has a certain degree of clinical value in the preoperative evaluation of HCC.

Objective:To investigate the effect of situational simulation combined with video feedback in cardiopulmonary and abdominal clinical probation of diagnostics.Methods:Two hundred and twenty-four clinical undergraduates from the second affiliated hospital of Guangxi Medical University were randomized into the experimental group ( n=112) and the control group ( n=112). The experimental group applied situational simulation combined with video feedback methods in the cardiopulmonary and abdominal clinical probation. In other words, according to the assignment of diseases, two students played the role of "physician and patient" and conducted corresponding consultation and physical examination, in which the positive and negative video feedback were integrated to standardize students' physical examination skills. The control group adopted conventional teaching method. The teaching effect of the two groups was evaluated through unified and standardized scores of theoretical tests, cases analysis, and clinical skills operation, and the experimental group was conducted anonymous questionnaires to 112 students. SPSS 22.0 was used to perform the t test. Results:The evaluation results show that the average scores of theoretical tests (90.12±0.94), cases analysis (9.30±0.10) and physical examination (93.50±0.72) in the experimental group are higher than those of the control group, (83.20±1.66), (8.10±0.11), (85.00±1.75), respectively, with statistical significance ( P<0.05). All 112 questionnaires were collected, with an effective rate of 100%. The percentage of "yes" respondents is as follows, 91.1% of "Enhancing the mastery and application of knowledge"; 97.3% of "Cultivating clinical thinking"; 87.5% of "Improving doctor-patient communication skills"; however, there are 93.8% students believe that this new teaching approach takes more time to prepare before class, consequently, their initiative and enthusiasm of study and learning interest have slid into decline, with percentage of 80.4% and 83.0%, respectively. Conclusion:The teaching method of situational simulation and video feedback in cardiopulmonary and abdominal clinical probation of diagnostics is beneficial to enhance students' understanding and mastery of theoretical knowledge, standardize physical examinations, cultivate clinical thinking ability and improve the doctor-patient communication skills. However, it takes a lot of time for students to prepare before class. If they fail to make reasonable arrangements, the teaching effect will be half the effort and students' initiative and enthusiasm of learning will be reduced.

Objective: To investigate the cellular and humoral immune responses induced by combined immunization with the fusion protein of human papillomavirus type 18 (HPV18) and the recombinant vaccinia virus. Methods: Purified HPV18L2_31-600E7E6 fusion protein, expressed by prokaryotic expression system, were immunized in combination with the recombinant vaccinia virus vaccine expressing HPV18E7E6 fusion protein (rVV18E7E6) by using various prime-boost regiments in C57BL/6 mice. Cellular and humoral immune responses were analyzed by enzyme-linked immunospot assay (ELISPOT), enzyme-linked immunosorbent assay (ELISA), and pseudovirus neutralization assay. Results: Higher levels of cellular immune responses were induced in mice primed with the HPV18L2_31-600E7E6 fusion protein/adjuvant CpG and boosted with the recombinant vaccinia virus rVV18E7E6, than in other immunized mice. Higher binding antibody level was induced, and low level neutralizing antibody against pseudovirus was detected simultaneously. Conclusions Priming with HPV18L2_31-600E7E6 fusion protein/CpG and boosting with the recombinant vaccinia virus rVV18E7E6 could induce higher cellular and humoral immune response in immunized mice, which might be taken as vaccine candidate for treatment of HPV18 chronic infection and postoperative adjuvant treatment for cervical cancer.