Electrochemical DNA biosensor for the multianalyte detection of HPV-16 and HPV-18 in oral and cervical lesions

Infection with high-risk human papillomavirus (HPV) is a major risk factor for oral and cervical cancers. In this study, we developed an electrochemical DNA biosensor for detection of HPV-16 and HPV-18, which are the 2 most prevalent genotypes for development of oral and cervical cancers. The assay involves the sandwich hybridization of the HPV target to silica-redox dye reporter probe and capture probe, followed by electrochemical detection. The sensor was found to be highly specific and sensitive, with detection limit of 22 fM for HPV-16 and 20 fM for HPV-18, between the range of 1 fM to 1 µM. Evaluation with oral and cervical samples showed that the biosensor result was consistent with the nested PCR /gel electrophoresis detection. The biosensor assay could be completed within 90 minutes. Due to its simplicity, rapidity and high sensitivity, this biosensor could be used as an alternative method for HPV detection in clinical laboratories. [151 words]

Identification and validation of the high expression of pseudogene TCAM1P in cervical cancer via integrated bioinformatics analysis

Background HPV as the main cause of cervical cancer has long been revealed, but the detailed mechanism has not yet been elucidated. The role of testis/cancer antigen in cervical cancer has been revealed. However, there are no reports about the statement of testis/cancer-specific non-coding RNA. In this study, we first proposed TCAM1P as a testis/cancer-specific pseudogene, and used a series of experimental data to verify its relationship with HPV, and analyzed its diagnosis value of high-grade cervical lesions and the mechanism of their high expression in cervical cancer. This provides a new direction for the prevention and treatment of cervical cancer. Methods The specific expression of pseudogenes in each tissue was calculated by “TAU” formula. ROC curve was used to judge the diagnosed value of TCAM1P for high-grade lesions. The proliferation ability of cells was measured by CCK8. The expression of TCAM1P, HPV E6/E7 were detected by qRT-PCR. The binding for RBPs on TCAM1P was predicted by starbase v2.0 database, then RIP assay was used to verify. Besides, Gene Ontology (GO) and KEGG enrichment analysis were performed with “clusterprofiler” R package. Results TCAM1P was specifically high-expressed in normal testicular tissue and cervical cancer. Interesting, with the severity of cervical lesions increased, the expression of TCAM1P increased, and TCAM1P could effectively diagnose high-grade cervical lesions. Besides, the expression of TCAM1P was HPV dependent, with highest expression in HPV-positive cervical cancer tissues. Furthermore, RIP assay showed that EIF4A3 regulated the expression of TCAM1P through binding with it. CCK8 assay showed that TCAM1P promoted the proliferation and the Gene ontology (GO) and KEGG Pathway enrichment analysis same suggested that TCAM1P is involved in multiple ways in cell proliferation including Cell cycle, DNA replication and etc. Conclusions In this study, we firstly proposed that TCAM1P is cancer/testis pseudogene and is regulated by HPV E6/E7 and EIF4A3. TCAM1P promotes the proliferation of cervical cancer cells and acts as promoter in cervical cancer. Otherwise, TCAM1P promote proliferation through regulating cell cycle and DNA replication, but more evidence needs to be provided to reveal the mechanism by which TCAM1P plays a role in cervical cancer.

Five-year Randomized Clinical Trial on the Performance of Two Etch-and-rinse Adhesives in Noncarious Cervical Lesions

SUMMARY Objectives: To evaluate the 5-year clinical performance of two-step etch-and-rinse adhesives in noncarious cervical lesions (NCCL). Methods and Materials: The sample comprised 35 adults with at least two similar-sized NCCL. Seventy restorations were placed, according to one of the following groups: Adper Single Bond 2 (SB) and Ambar (AM). The restorations were placed incrementally using a resin composite (Opallis). The restorations were evaluated at baseline and after 6 and 18 months and 5 years using some items of the FDI criteria. The differences in the ratings of the two materials after 6 months, 18 months, and 5 years were performed with Friedman repeated measures ANOVA by rank and McNemar test for significance in each pair (α=0.05). Results: Five patients did not attend the 60-month recall. No significant differences were observed between the materials for any criteria evaluated. Twenty-one restorations failed (12 for SB and 9 for AM) after 60 months. Thus, the retention rate for SB at 60 months were 55.6% for SB and 71% for AM (p=0.32). After 60 months, 12 restorations (6 for SB and 6 AM) showed some loss of marginal adaptation (p=1.0). Slight marginal discoloration was observed in 10 restorations (6 for SB and 4 AM; p=0.91). Five restorations (2 for SB and 3 for AM) showed recurrences of caries (p=1.0). Conclusions: Both two-step etch-and-rinse adhesives—Adper Single Bond 2, a polyalkenoic acid-containing adhesive, and Ambar, a 10-methacryloyloxydecyl dihydrogen phosphate (MDP)-containing adhesive—showed acceptable clinical performance after 60 months.

Evaluation of Dynamic Thiol-Disulfide Homeostasis on Hpv Positive-Women in Progression to Cervical Intraepithelial Lesion

PURPOSE: Dynamic thiol disulfide homeostasis (TDH) is critical in cervical carcinogenesis at HPV infection as a sign of antioxidant consumption native and total thiol levels decrease in progress to cervical intraepithelial lesions. TDH is the main actor in signaling pathways, apoptosis, antioxidant and detoxification reactions. In this study, we aimed to evaluate the effect of TDH intraepithelial progression of cervical precancerous lesions on HPV positive women.METHODS: This was a prospective cross-sectional study. Subjects were selected from newly diagnosed high risk HPV DNA-positive patients. TDH results were calculated as the levels of disulfide, native and total thiol, the ratios of disulfide/total thiol (SS/SH+SS), disulfide/native thiol (SS/SH) and native thiol/total thiol (SH/SH+SS).RESULTS: A total of 146 women were included in the study. Study groups were as group one; control included 66 participants, group two; HPV DNA-positive women without preinvasive cervical lesion included 30 participants and group three; HPV DNA-positive women with preinvasive cervical lesion included 50 participants. Native and total thiol levels were elevated on HPV-positive women without preinvasive cervical lesions. There were no significant differences between groups related to the ratios of SS/SH, SS/ Total SH, SH/ Total SH levels. CONCLUSIONS: HPV infection related to oxidative stress has effects on oxidant/antioxidant balance and could be demonstrated in systemic circulation by TDH parameters. Consumption of thiol substances play role in the cervical neoplastic process, replacement with antioxidants would be a treatment option for HPV infections.

Ultrasonic Elastography Combined with Human Papilloma Virus Detection Based on Intelligent Denoising Algorithm in Diagnosis of Cervical Intraepithelial Neoplasia

The aim of this research was to study the application of ultrasonic elastography combined with human papilloma virus (HPV) detection based on bilateral filter intelligent denoising algorithm in the diagnosis of cervical intraepithelial neoplasia (CIN) and provide a theoretical basis for clinical diagnosis and treatment of CIN. In this study, 100 patients with cervical lesions were selected as research objects and randomly divided into control group and experimental group, with 50 cases in each group. Patients in control group and experimental group were diagnosed by ultrasonic elastography combined with HPV detection. The experimental group used the optimized image map of bilateral filter intelligent denoising algorithm for denoising and optimization, while the control group did not use optimization, and the differences between them were analyzed and compared. The diagnostic effects of the two groups were compared. As a result, the three accuracy rates of the experimental group were 95%, 95%, and 98%, respectively; the three sensitivity rates were 96%, 92%, and 94%, respectively; and the three specificity rates were 99%, 97%, and 98%, respectively. In the control group, the three accuracy rates were 84%, 86%, and 84%, respectively; the three sensitivity rates were 88%, 84%, and 86%, respectively; and the three specificity rates were 81%, 83%, and 88%, respectively. The accuracy, sensitivity, and specificity of experiment group were significantly higher than those of control group, and the difference was statistically significant ( P < 0.05 ). In summary, the bilateral filter intelligent denoising algorithm has a good denoising effect on the ultrasonic elastography. The ultrasonic image processed by the algorithm combined with HPV detection has a better diagnosis of CIN.

Universal Adhesives and Adhesion Modes in Non-Carious Cervical Restorations: 2-Year Randomised Clinical Trial

This prospective, double-blind, six-arm parallel randomised controlled trial aimed to compare the performance of two universal adhesives (UAs) in non-carious cervical lesions (NCCLs), using the FDI criteria, and analysed if participants/NCCLs’ characteristics influenced the outcome. Thirty-eight 18- to 65-year-old participants were seeking routine dental care at a university clinic. At baseline, 210 NCCLs were randomly allocated to six groups (35 restorations’ each). The UAs tested were FuturabondU (FBU) and AdheseUniversal (ADU) applied in either etch-and-rinse (ER) and self-etch (SE) modes. FuturabondDC (FBDC) in SE and in SE with selective enamel etching (SE-EE) modes were controls. NCCLs were restored with AdmiraFusion. The analysis included nonparametric tests, Kaplan-Meier and log-rank tests (α = 0.05). At 2-years, of 191 restorations, ten were missed due to retention loss (all groups, p > 0.05). FBDC (p = 0.037) and FBU (p = 0.041) performed worse than ADU in SE mode. FBDC and FBU also showed worse functional success rate (p = 0.012, p = 0.007, respectively) and cumulative retention rates (p = 0.022, p = 0.012, respectively) than ADU. Some participants/NCCLs’ characteristics influenced (p < 0.05) the outcomes. FBU did not perform as well as ADU, especially in SE mode and due to functional properties. Participants’ age and NCCLs’ degree of dentin sclerosis and internal shape angle influenced FBU performance.

Serum Tie-1 is a Valuable Marker for Predicting the Progression and Prognosis of Cervical Cancer

Objective: To investigate whether serum Tie-1 (sTie-1) is a valuable marker for predicting progression and prognosis of cervical cancer.Methods: Enzyme-linked immunosorbent assay (ELISA) was used to detect serum sTie-1 concentrations in 75 cervical cancer patients, 40 cervical intraepithelial neoplasia (CIN) patients, and 55 healthy controls without cervical lesions, and sTie-1 levels were compared between the groups. Receiver operating characteristic curves was used to evaluate the diagnostic value of sTie-1. The relationship between sTie-1 concentrations in patients with cervical cancer and clinicopathological features and prognosis were analyzed, and the risk factors for postoperative recurrence were determined using univariate and multivariable Cox proportional hazards regression.Results: We found that sTie-1 concentrations gradually increased according to lesion severity (i.e., cancer vs. CIN; p &lt; 0.05) and were significantly elevated in adenocarcinoma compared with healthy controls. sTie-1 levels strongly distinguished between cervical cancer patients and the healthy controls (area under the curve = 0.846; cut-off value = 1,882.64 pg/ml; sensitivity = 74.6%; specificity = 96.4%). Moreover, sTie-1 levels in cervical cancer patients were significantly associated with tumor size, advanced tumor stage, lymph node metastasis, and reduced 4-years progression-free survival. Cervical cancer patients with high sTie-1 concentrations had a 3.123-fold [95% confidence interval (CI): 1.087–8.971, p = 0.034] higher risk for tumor recurrence.Conclusions: Elevated sTie-1 levels in patients with cervical carcinoma were associated with tumor progression and poor prognosis, indicating that sTie-1 may be a valuable marker for predicting progression and prognosis of cervical cancer.