Abstract Background At present, there are certain deficiencies in cervical cancer screening methods. Consequently, we aimed to establish a prediction model for cervical precancerous lesions utilizing DNA methylation haplotype load (MHL) markers. Methods Three machine learning models, namely, a random forest model (RF), a support vector machine model (SVM), and a naive Bayes model (NB), were developed based on the performance of 47 regions in 52 HR-HPV + cervical cytology specimens. Additionally, external validation of the three models was conducted using 101 HR-HPV + cervical cytological samples. Results From the initial 52 samples, we derived three models with respective area under the curve (AUC) values of 0.864, 0.867 and 0.847. Subsequently, in the validation phase involving 101 samples, the RF model demonstrated superior performance compared to the E6 protein detection model and p16Ki67 model in predicting cervical intraepithelial neoplasia grade 2 (CIN2) lesions and above. Compared with the human papillomavirus (HPV) combined with cytology triage model, the RF prediction model had a PPV of 100% and an NPV of 95.15% in the HPV16/18-positive subgroup. Within the other high-risk-HPV-positive subgroups, the PPV and NPV were 41.67% and 92.36%, respectively. Conclusions Our findings suggest that the methylation haplotype load markers of multiple genes offer clear advantages in screening CIN2 lesions and above in cervical cancer. Furthermore, enhancing the methylation detection method has the potential to further improve the NPV of the prediction model.

The SPF10‐polymerase chain reaction (PCR)‐based line probe assay (LiPA‐25) with high analytical sensitivity and specificity for human papillomavirus (HPV) genotyping in clinical samples has been widely used in vaccine and epidemiologic studies. A real‐time multiplex PCR assay using type‐specific primers (Hybribio‐23) with low workload and cost has been developed recently. The study aimed to compare the performance of LiPA‐25 and Hybribio‐23 in selected 1731 cervical swab and 117 tissue samples, with a focus on 20 common HPV types (14 high‐risk: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68/73; 6 low‐risk: 6, 11, 42, 43, 44, and 53). The level of agreement of two assays was determined using Cohen's Kappa ( κ ) statistics. A total of 1296 (74.9%) swab samples were identified as HPV‐positive by Hybribio‐23 or LiPA‐25, of which 814 (62.8%) samples exhibited concordant, 358 (27.6%) showed additional or fewer types (compatible), and 124 (9.6%) were discordant. In addition, the two assays showed a perfect agreement for 20 HPV‐combined detection ( κ = 0.838) and 17 individual HPV types (all κ > 0.800), a good agreement for HPV31 ( κ = 0.792) and 43 ( κ = 0.696), and a moderate agreement for HPV42 ( κ = 0.504). Hybribio‐23 was significantly more sensitive for HPV58, 59, 68/73, 42, 43, and 44, and less sensitive for HPV35 and 66 than LiPA‐25 (McNemar's test: all p < 0.05). For 117 HPV‐positive tissue specimens, the identification of genotypes was 85.2% identical, 12.2% compatible, and only 2.6% discordant. The agreement for HPV31 ( κ = 0.786), 68/73 ( κ = 0.742), and HPV53 ( κ = 0.742) was good, while for other types (all κ > 0.853) and 20 HPV‐combined detection ( κ = 0.936) was perfect (all p > 0.05). In conclusion, Hybribio‐23 and LiPA‐25 are comparable. Hybribio‐23 could be used for the detection and genotyping of HPV in cervical samples for epidemiological and vaccine studies worldwide.

The SPF10‐PCR‐based line probe assay (LiPA‐25) for human papillomavirus (HPV) genotyping with high analytical sensitivity and specificity was widely used in HPV vaccine clinical trials and epidemiologic studies. In the study, we aimed to compare a novel GP5+/6+‐PCR‐based reverse dot blot hybridization assay (Yaneng‐23) with LiPA‐25. The performance of two assays was evaluated in 1735 cervical swab and 117 tissue samples, with a focus on 19 common HPV types (14 high‐risk: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68/73; 5 low‐risk: 6, 11, 42, 43, and 53). A total of 1197 (69.0%) swab samples were identified as HPV‐positive by two assays. Of these, 878 (73.4%) samples displayed absolute agreement (concordant), 255 (21.3%) showed additional or fewer types (compatible), and the remaining 64 (5.3%) samples were discordant. Additionally, the two assays showed an excellent strength of agreement for 19 HPV‐combined detection (κ = 0.886) and 17 individual HPV types (all κ > 0.800), and displayed a good agreement for HPV39 (κ = 0.780) and 42 (κ = 0.699). Yaneng‐23 was more sensitive than LiPA‐25 for HPV58, 59, 68/73, 42, 43 and 53 (McNemar's test: all p < 0.05), while LiPA‐25 was more sensitive for HPV31, 39, 52, and 66 than Yaneng‐23 (all p < 0.05). In 113 HPV‐positive tissue specimens, the identification of genotypes was 82.3% identical and 17.7% compatible. The agreement between the tests for HPV45 (κ = 0.796) and 51 (κ = 0.742) was good, and for other types (all κ > 0.843) and 19 HPV‐combined detection (κ = 0.929) was perfect (all p > 0.05). In conclusion, Yaneng‐23 and LiPA‐25 are comparable. Yaneng‐23 could be used for the detection and genotyping of HPV in cervical samples for epidemiological and vaccine studies worldwide.

Background The causal role of high-risk Human papillomavirus (HR-HPV) in the pathogenesis of anogenital cancers is well established. In contrast, information on HR-HPV distribution of continuous anatomic sites within the female genital tract is limited, and the impact of sample type on the clinical performance in HPV-based cervical cancer screening warrants investigation. Methods A total of 2,646 Chinese women were enrolled in the study from May 2006 to April 2007. We analyzed the infection features by infection status and pathological diagnoses of 489 women with complete HR-HPV type and viral load data on the cervix, upper vagina, lower vagina, and perineum samples. Additionally, we assessed the clinical performance for detecting high-grade cervical intraepithelial neoplasia of grade two or worse (≥ CIN2) among these four types of samples. Results HR-HPV positivity rate was lower in the cervix (51.53%) and perineum (55.83%), higher in the upper (65.64%) and lower vagina (64.42%), and increased with the severity of cervical histological lesions (all P<0.001). Single infection was more dominant than multiple infections at each anatomic site of the female genital tract. The proportion of single HR-HPV infection decreased successively from the cervix (67.05%) to the perineum (50.00%) ( P trend =0.019) in cervical intraepithelial neoplasia grade 1 (CIN1) and was higher in samples of the cervix (85.11%) and perineum (72.34%) in ≥ CIN2. In addition, the highest viral load was observed in the cervix compared to the other three sites. The overall agreement of the cervical and perineum samples was 79.35% and increased continuously from normal (76.55%) to ≥ CIN2 (91.49%). As for the detection of ≥ CIN2, the sensitivity was 100.00%, 97.87%, 95.74%, and 91.49% for the cervix, upper vagina, lower vagina, and perineum samples, respectively. Conclusions Single HR-HPV infection predominated throughout the female genital tract, but the viral load was lower compared to multiple HR-HPV infections. Despite the decreasing viral load from cervix to perineum, the clinical performance for detecting ≥ CIN2 of the perineum sample was comparable to that of the cervix.

Background Studies on the clinical performance of p16/Ki-67 dual-staining in detecting cervical lesions by menopausal status were limited. Methods 4364 eligible women were enrolled with valid p16/Ki-67, HR-HPV, and LBC test results, including 542 cancer and 217 CIN2/3 cases. The positivity rates of p16 and Ki-67 single staining and p16/ Ki-67 dual-staining were analyzed by different pathological grades and age groups. The sensitivity (SEN), specificity (SPE), positive predictive value (PPV), and negative predictive value (NPV) of each test in different subgroups were calculated and compared. Results P16/Ki-67 dual-staining positivity increased with histopathological severity in premenopausal and postmenopausal women ( P < 0.05), while no increasing trends of individual expression of p16 single staining and Ki-67 single staining were observed in postmenopausal women. P16/Ki-67 showed higher SPE (88.09% vs. 81.91%, P < 0.001) and PPV (33.8% vs. 13.18%, P < 0.001) in detecting CIN2/3, and higher SEN (89.97% vs. 82.61%, P = 0.012) and SPE (83.22% vs. 79.89%, P = 0.011) in detecting cancer in premenopausal women than postmenopausal women. For triaging the HR-HPV+ population to identify CIN2/3, p16/Ki-67 performed comparably to LBC in the premenopausal women, and showed higher PPV (51.14% vs. 23.08%, P < 0.001) in premenopausal than postmenopausal women. For triaging ASC-US/LSIL population, p16/Ki-67 demonstrated higher SPE and lower colposcopy referral rate than HR-HPV in both premenopausal and postmenopausal women. Conclusions Expressions of p16/Ki-67 dual-staining between premenopausal and postmenopausal women are varied. P16/Ki-67 performs better in detecting cervical lesions in premenopausal women. For triaging, p16/Ki-67 is suitable for HR-HPV+ women, especially premenopausal women, to identify CIN2/3 and women with ASC-US/LSIL.

OBJECTIVE:To assess the association between endocervical adenocarcinoma (ECA) and HPV (Human papillomavirus) infection, as well as the characteristics of ECA distribution in China. METHODS:A total of 756 specimens were collected from seven geographic regions across China. All cases were histologically categorized according to the 2020 WHO classification of female genital tract cancers, and 496 cases were included. We performed the SPF10-DEIA-LiPA25 assay on all specimens' whole tissue sections using PCR (WTS-PCR) to detect HPV DNA and 141 WTS-PCR HPV-positive specimens were selected for the laser capture microdissection (LCM). RESULTS:Four predominant prevalent histological categories of ECA in China were usual type (51.8%, 257), invasive stratified mucin-producing carcinoma (iSMILE) (11.5%, 57), mucinous NOS (not otherwise specid) (10.3%, 51), and gastric type (7.9%, 39). HPV positivity was 91.4% (235/257), 100.0% (57/57), and 90.2% (46/51) in usual type, iSMILE, and mucinous NOS by WTS-PCR detection, respectively (P < 0.001). LCM-PCR results showed a decreasing trend in HPV DNA positivity, and 21 (95.5%) patients with HPV-I were negative for HPV-DNA in glandular epithelial tissue. The most prevalent HPV genotypes in ECA were HPV16 (47.5%), 18 (40.8%), and 52 (6.5%). The average age of patients with HPVA was 44.9 years, while that of patients with HPV-I was 49.1 years, HPVA is more prevalent in younger females in China (P < 0.001). CONCLUSIONS:In China, the predominant prevalent histological category of ECA is the usual type of adenocarcinoma, followed by iSMILE. Additionally, patients with HPVA tended to be younger than those with HPV-I.

Background: Quadruple therapy with polaprezinc provided an alternative to Helicobacter pylori eradication; however, the effect on gut microbiota remains uncertain. This study aims to identify whether polaprezinc-containing quadruple therapy causes adverse microbiota effects among asymptomatic adults, compared with bismuth therapy. Methods: This was a randomized control trial. One hundred asymptomatic H. pylori-infected adults were randomly (1:1) assigned to two treatment groups (polaprezinc-containing therapy, PQT; or bismuth-containing therapy, BQT). Fecal samples were collected from subjects before and 4–8 weeks after therapy. Samples were sequenced for the V4 regions of the 16S rRNA gene. Results: The relative abundance of the three dominant bacterial phyla (Bacteroidota, Firmicutes, and Proteobacteria) accounted for more than 95% of each treatment group. The alpha diversity between eradications that succeeded and those that failed had no significant difference (p > 0.05). After successful eradication, the alpha diversity in the BQT group decreased in comparison with the baseline (p < 0.05). Subjects who were successfully eradicated by BQT showed considerably lower alpha diversity indices than those of the PQT at follow-up (p < 0.05). The abundance of Parasutterella in subjects who were successfully eradicated by PQT was four times greater than that of BQT (q < 0.05). Conclusion: A 14-day PQT may be superior to BQT in maintaining short-term gut microbiota homeostasis after H. pylori treatment. Our findings preliminarily provide evidence of the short-term impacts of the gut microbiota after PQT treatment of H. pylori infection.