Objective: To develop a therapeutic adjuvantfree protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: First the E6/E7 gene by PCR technology from HPV16z virus strain was isolated in the highrisk cervical cancer area of Shanxi province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28aHsp65E6/E7, expressed the Hsp65E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvantfree protein vaccine——Hsp65E6/E7.

Objective To investigate the heterogeneity of chemosensitivity in colorectal cancer using an ATP-tumor chemosensitivity assay (ATP-TCA) and the feasibility of individual chemotherapy.Methods An ATP-TCA were used to determine the effect of 16 single or combined cytotoxic drugs in surgical specimens from 50 patients with colorectal cancer.Results There were considerable differences in chemosensitivity between individuals.The most active single drugs in the assay was identified as Navelbine, Hydroxycamptothecin, 5-Fluorouracil and Paclitaxel; 34.1%, 31.6%, 27.6% and 24.3% of specimens showed sensitivity to them, respectively.5-Fluorouracil+Mitomycin+Aytarabine was found to be the most effective combination, for 100% (11/11)specimens were sensitive to this regimen.5-Fluorouracil+Cisplatin+Adriamycin and Gemcitabine+Cisplatin were moderately active regimens.Conclusions There was the heterogeneity of the in vitro chemosensitivity in colorectal cancer.The use of the ATP-TCA provides a method of selecting appropriate anti-cancer drugs in colorectal cancer.

Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response, and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo and the influence on mouse lifetime. Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could proliferate obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes in vitro. This animal therapeutic experiment in vivo showed that the vaccine suppressed the growth of TC-1 cell transplanted tumor remarkably. Conclusion: The recombined vaccine can induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.