Application Value of Detection of High-Risk HPV Infection in Early Cervical Cancer Patients in Disease Diagnosis and Prognosis Evaluation.

BACKGROUND: To analyze the relationship between HPV infection and early cervical cancer and postoperative survival outcomes. METHODS: A total of 556 women were recruited to receive TCT and HPV tests from October 2017 to October 2018. The type of disease was pathologically diagnosed. The HPV positive rate, HPV-DNA, and E6/E7 mRNA quantitative level were detected, and the diagnostic accuracy of the subjects was analyzed by the receiver operating characteristic (ROC). The cervical intraepithelial neoplasia (CIN) and early cervical cancer patients were radically cured and followed up for 12.0 months to analyze the recurrence rate. RESULTS: Seventy-two cases of chronic cervicitis, 54 cases of CIN, and 51 cases of cervical cancer patients were pathologically diagnosed (32 cases in early stage and 19 cases in middle and late stage). HPV positive rate increased gradually in chronic cervicitis, CIN, and cervical cancer group (p < 0.001) and HPV 16 + 18 subtype. The positive rate was significantly different (p = 0.009). HPV-DNA and E6/E7 mRNA quantification also showed significant differences (p < 0.001). ROC analysis indicated that the accuracy of HPV-DNA and E6/E7 mRNA quantitative diagnosis of malignant lesions (CIN+ cervical cancer) were 0.865 and 0.879, respectively. There were 4 cases (7.41%) of recurrence in CIN group and 5 cases (15.63%) in early cervical cancer group. There was no difference (p = 0.401) among all of the patients. All patients with recurrence were HPV positive. CONCLUSIONS: HPV detection is an indispensable screening method for early cervical cancer and precancerous lesions, and comprehensive HPV 16 and 18 subtypes. DNA and E6/E7 mRNA quantification assay would further improve the accuracy of screening.

Evaluation of hGM-CSF/hTNFα surface-modified prostate cancer therapeutic vaccine in the huPBL-SCID chimeric mouse model.

To validate its efficacy in the context of the human immune system, a novel therapeutic vaccine of hGM-CSF/hTNFα surface-modified PC-3 cells against human prostate cancer was evaluated in the human peripheral blood lymphocytes-severe combined immunodeficiency (huPBL-SCID) chimeric mouse model. The hGM-CSF or/and hTNFα modified vaccines inhibited prostate cancer growth effectively so as to prolong the mouse survival significantly. The splenocytes from the hGM-CSF/hTNFα vaccine-inoculated mice showed the strongest tumor-specific cytotoxicity against PC-3 cells and the highest production of IFNɤ. These features indicated that type 1 protective immune response was induced efficiently against human prostate cancer and further enhanced through synergetic adjuvant effects of hGM-CSF and hTNFα.

Efficient production of sTNFRII-gAD fusion protein in large quantity by use of the modified CHO-S cell expression system.

TNFα is one of the initial and important mediators to activate downstream signaling pathways by binding to trimerized TNFα receptors (TNFR), and thus is an ideal drug target for cancer therapy. Taking advantage of intrinsic homotimerization of the globular domain of adiponectin (gAD), we have developed a novel TNFα antagonist, the trimerized fusion protein named sTNFRII-gAD. However, our previously-used CHO expression system yielded less than 10 mg/L of sTNFRII-gAD. To produce large quantities of sTNFRII-gAD efficiently, we used a modified CHO-S cell expression system, which is based on a pMH3 vector with non-coding GC-rich DNA fragments for high-level gene expression. We obtained stable clones that produced 75 mg/L of sTNFRII-gAD in the 96-well plate, adapted the clones to 40 ml suspension serum-free batch culture, then optimized the culturing conditions to scale up the fed-batch culture in a 3 L shake-flask and finally in a 5 L AP30 bioreactor. We achieved a final yield of 52 mg/L of sTNFRII-gAD. The trimerized sTNFRII-gAD exhibited the higher affinity to TNFα with a dissociation constant (Kd) of 5.63 nM than the dimerized sTNFRII-Fc with a Kd of 13.4 nM, and further displayed the higher TNFα-neutralizing activity than sTNFRII-Fc (p<0.05) in a L929 cytotoxicity assay. Therefore, the strategy employed in this study may provide an efficient avenue for large-scale production of other recombinant proteins by use of the modified CHO-S cell expression system.