[Clinical significance of HPV L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women].

OBJECTIVE: To explore the clinical significance of human papillomavirus L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women. METHODS: From November 2012 to June 2013, 386 high-risk HPV positive (detected by hybrid capture II) cases were enrolled as eligible women from Huzhou Maternity & Child Care Hospital and Women's Hospital, School of Medicine, Zhejiang University. All eligible women underwent liquid-based cytology (ThinPrep) followed by colposcopy. Biopsies were taken if indicated. Cervical exfoliated cells were collected for HPV L1 capsid protein detection by immunocytochemistry. Expression of HPV L1 capsid protein in groups with different histological diagnosis were compared, and the role of HPV L1 capsid protein detection in cervical exfoliated cells in cervical lesions screening was accessed. RESULTS: Total 386 enrolled eligible women were finally diagnosed histologically as follwed: 162 normal cervix, 94 low-grade squamous intraepithelial lesion (LSIL), 128 high-grade squamous intraepithelial lesion (HSIL) and 2 squamous cervical cancer (SCC). The positive expression rate of HPV L1 in HSIL+ (HSIL or worse) group was significantly lower than that in LSIL- (LSIL or better) group (19.2% vs 66.4%, P=0.000). While identifying HSIL+ in HPV positive cases and compared with cytology, HPV L1 detection resulted in significant higher sensitivity (80.77% vs 50.77%, P=0.000) and negative predictive value (NPV; 87.18% vs 76.47%, P=0.004), significant lower specificity (66.41% vs 81.25%, P=0.000), and comparable positive predictive value (PPV; 54.97% vs 57.89%, P=0.619). To identify HSIL+ in HPV-positive/cytology-negative women, the sensitivity, specificity, PPV, and NPV of HPV L1 detection were 87.50%, 61.54%, 41.18%, and 94.12% respectively, while 80.00%, 86.36%, 80.00% and 86.36% respectively in HPV-positive/atypical squamous cell of undetermined significance (ASCUS) women. CONCLUSIONS: HPV L1 capsid detection in cervical exfoliated cells have a role in cervical lesions screening in high-risk HPV positive women, and may be a promising triage for high-risk HPV-positive/cytology-negative or ASCUS women.

The Value of MicroRNA-375 Detection for Triaging Primary Human Papillomavirus Positive Women: A Cross-Sectional Study in a General Population.

PURPOSE: This study aims to validate the value of microRNA (miRNA) detection for triaging human papillomavirus (HPV)-positive women in the general population. PATIENTS AND METHODS: miR-375 detection in cervical exfoliated cells has been demonstrated to have the superior value to cytology in triaging primary HPV-positive women in the hospital population. In this study, residual samples of cervical exfoliated cells from 10,951 women in a general population were used to detect miRNA. The performance efficiency of miRNA detection in identifying high-grade cervical intraepithelial neoplasia (CIN) was evaluated. Pearson chi-square test and McNemar pairing test were used to compare miRNA detection and cytology. RESULTS: In valid 9,972 women aged 25-65, miR-375 expression showed a downward trend along with an increase in cervical lesion severity. The expression level of miR-375 ≤1.0 × 10-3 was identified as positive. In the HPV-positive and 12 HPV genotypes other than 16/18 (HR12)-positive women, miR-375 detection showed equivalent sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) to that of cytology (≥ASC-US) and higher or similar sensitivity and NPV but lower specificity and PPV than that of cytology (≥ASC-H) in identifying CIN3+ and CIN2+. In HPV 16-positive women, miR-375 positivity had higher sensitivity and NPV but lower specificity and PPV than that of cytology (≥ASC-H and HSIL) in identifying CIN3+ and CIN2+. The immediate CIN3+ risk of miR-375 positivity was 19.8% (61/308) in HPV-positive, 10.8% (22/204) in HR12-positive, and 43.5% (37/85) in HPV16-positive women, respectively. CONCLUSION: The detection of miR-375 in cervical exfoliated cells may be an optional method for triaging primary HPV-positive women in population-based cervical cancer screening.

The nonsynonymous single-nucleotide polymorphisms in codon 31 of p21 gene and the susceptibility to cervical cancer in Chinese women.

BACKGROUND: It was suggested that single-nucleotide polymorphisms in p21 codon 31 seem to be associated with a variety of human malignancies; very few studies have focused on the association between p21 codon 31 polymorphisms and cervical cancer. This study explored whether p21 codon 31 nonsynonymous single-nucleotide polymorphisms might be associated with an increased risk of cervical cancer development among Chinese women. METHODS: Peripheral blood samples were obtained from patients with cervical cancer (n = 317) and healthy controls (n = 353) for detecting the biallelic polymorphisms at codon 31 of p21 gene by the mismatch amplification mutation assay-polymerase chain reaction. Cervix brush-off samples were obtained from patients with cervical squamous cell carcinoma (SCC) and controls for detection of high-risk human papillomavirus (HR-HPV). RESULTS: The AGA (Arg) allele frequency in patients with cervical SCCs was significantly higher than that in controls. AGA/AGA and AGA/AGC genotypes were more frequently found in cervical SCCs than in controls. There was no significant difference of allele frequency or genotype distribution between cervical adenocarcinomas and controls, or between HR-HPV-positive and HR-HPV-negative groups. CONCLUSIONS: p21 Codon 31 with AGA (Arg) allele is a genetic risk factor of cervical SCC, and the increased risk is probably not caused by increasing host susceptibility to HR-HPV infection.

[Analysis of the relationship between clinicopathological features and pelvic lymph node metastasis in patients with early stage squamous cell carcinoma of the uterine cervix].

OBJECTIVE: To evaluate clinical and pathologic factors associated with pelvic lymph node metastasis in patients with early-stage squamous cell carcinoma of the uterine cervix. METHODS: From February 2004 to January 2007, 135 patients with stage Ib-IIa cervical squamous cell carcinoma in Women's Hospital, School of Medicine, Zhejiang University, were retrospectively studied. The relationship between pelvic lymph node metastasis and age, clinical stage, tumor size, grade of differentiation, depth of muscular invasion, lymphatic vascular space invasion, pretreatment level of serum squamous cell carcinoma antigen, pretreatment plasma level of fibrinogen, pretreatment levels of hemoglobin and platelet were evaluated by univariate and multivariate analyses. RESULTS: Totally 3996 lymph nodes were dissected in 135 patients, with an average of 29.6 lymph nodes in each patient. 12.6% of the patients (17/135) had metastasized pelvic lymph nodes. Univariate analysis indicated that tumor size (P = 0.003), depth of muscular invasion (P = 0.004), vascular space invasion (P < 0.01), pretreatment levels of platelet (P = 0.006) and fibrinogen (P < 0.01) were significantly related to pelvic lymph node metastasis. Multivariate logistic regression analysis showed that lymphatic vascular space invasion (OR: 3.674, 95% CI: 1.825 - 7.393, P < 0.01) and pretreatment plasma level of fibrinogen (OR: 4.568, 95% CI: 1.779 - 11.725, P = 0.002) were significantly related to pelvic lymph node metastasis in patients with early-stage squamous cell carcinoma of the uterine cervix. CONCLUSION: In early-stage cervical squamous cell carcinoma, lymphatic vascular space invasion and higher pretreatment plasma levels of fibrinogen are risk factors of pelvic lymph node metastasis.

Postmenopausal spontaneous rupture of pyometra: A case report.

RATIONALE: The incidence of spontaneous perforations in pyometra occurs rarely, only 0.01% to 0.5% in gynecological patients, with high mortality and morbidity. The clinical manifestation of perforated uterine pus is similar to that of gastrointestinal perforation, but the gynecological symptoms are not so obvious, which makes preoperative diagnosis difficult. Here, we report a rare case of peritonitis with laparotomy of pyometra. PATIENT CONCERNS: An acute abdominal pain and purulent vaginal discharge developed in a 72-year-old woman who underwent an emergency laparotomy because of signs of diffuse peritonitis and in a state of shock. DIAGNOSES: We made a diagnosis of spontaneous perforation of pyometra. INTERVENTIONS: At laparotomy, about 1000 mL of pus with the source of uterine was found in the abdominal cavity, while gastrointestinal tract was intact and a crevasse with a diameter of 1.5 cm on posterior uterine wall was obvious. A total abdominal hysterectomy and a bilateral salphingo oophorectomy were performed. OUTCOMES: The patient got discharged on 34th postoperative hospitalization day with only 1 complication of wound healing. Histopathological study revealed uterine purulent inflammation, with no evidence of malignancy. LESSONS: Ultrasonography is the first and most sensitive examination for the evaluation of pyometra, but has limited role in the diagnosis of perforated pyometra. Additional diagnostic radiographic evaluation use for acute abdomen is total abdomen computed tomography scan and magnetic resonance imaging techniques of female pelvis.

Human papillomavirus (HPV) E6/E7 mRNA detection in cervical exfoliated cells: a potential triage for HPV-positive women.

Cytology triage has been generally recommended for human papillomavirus (HPV)-positive women, but is highly dependent on well-trained cytologists. The present study was designed to explore whether HPV E6/E7 mRNA detection in cervical exfoliated cells can be a potential triage for HPV-positive women from a clinic-based population. Both the primary HPV testing and Papanicolaou (Pap) test were performed on all eligible HPV-positive women. HPV E6/E7 mRNA was detected by QuantiVirus® HPV E6/E7 mRNA assay in cervical exfoliated cells. All HPV-positive women underwent colposcopy and further biopsy if indicated. The data were assessed by Pearson's Chi-squared test and the receiver operating characteristic curve. A total of 404 eligible HPV-positive women were enrolled. Positive rate of E6/E7 mRNA in high-grade squamous intraepithelial lesion (HSIL) cases was higher than that in low-grade squamous intraepithelial lesion (LSIL) or normal cases. There was no statistical difference found between mRNA and cytological testing with sensitivity (89.52% vs. 86.67%, P=0.671), specificity (48.96% vs. 48.96%, P=1.000), positive predictive value (39.00% vs. 38.24%, P=1.000), and negative predictive value (92.76% vs. 90.97%, P=0.678) for detecting ≥HSIL. HPV E6/E7 mRNA detection in cervical exfoliated cells shows the same performance as Pap triage for HSIL identification for HPV-positive women. Detection of HPV E6/E7 mRNA may be used as a new triage option for HPV-positive women.

Whole-exome sequencing reveals genetic variants in ERC1 and KCNG4 associated with complete hydatidiform mole in Chinese Han women.

Complete hydatidiform mole (CHM) is a rare pregnancy-related disease with invasive potential. The genetics underlying the sporadic form of CHM have not been addressed previously, but maternal genetic variants may be involved in biparental CHM. We performed whole-exome sequencing of 51 patients with CHM and 47 healthy women to identify genetic variants associated with CHM. In addition, candidate variants were analyzed using single base extension and Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry in 199 CHM patients and 400 healthy controls. We validated candidate variants using Sanger sequencing in 250 cases and 652 controls, including 205 new controls. Two single nucleotide polymorphisms, c.G48C(p.Q16H) inERC1 and c.G1114A(p.G372S) in KCNG4, were associated with an increased risk of CHM (p<0.05). These variants may contribute to the pathogenesis of CHM and could be used to screen pregnant women for this genetic abnormality.

Early stage primary ovarian mucinous carcinoma: Outcome-based clinicopathological study in comparison with serous carcinoma.

OBJECTIVE: To compare clinicopathological characteristics and survival rates between patients with primary ovarian mucinous carcinoma and those with primary ovarian serous carcinoma. METHODS: This retrospective study reviewed archival tumour specimens, originally diagnosed as primary ovarian mucinous carcinoma, using refined histological criteria. All patients were contacted to establish survival status. Clinicopathological characteristics and patient survival data were compared with a group of control patients with primary ovarian serous carcinoma. RESULTS: Of the 33 patients originally diagnosed with primary ovarian mucinous carcinoma, this diagnosis was only confirmed in 18. Primary ovarian mucinous carcinoma was more commonly associated with early International Federation of Gynecology and Obstetrics tumour stages and low-grade histology than primary ovarian serous carcinoma. Patients with primary ovarian mucinous carcinoma had a significantly higher overall 5-year survival rate than those with primary ovarian serous carcinoma (12/12 [100%] versus 14/24 [58%]). Kaplan-Meier survival plots demonstrated that patients with primary ovarian mucinous carcinoma had a survival advantage over patients with primary ovarian serous carcinoma. CONCLUSIONS: Primary ovarian mucinous carcinomas are frequently low-grade, stage I tumours and have an excellent prognosis.

CUL2 overexpression driven by CUL2/E2F1/miR-424 regulatory loop promotes HPV16 E7 induced cervical carcinogenesis.

It has been shown that HPV16 E7, but not other genotypes, can bind to scaffold protein CUL2 during inducing cervical carcinogenesis, but the expression level, associated regulating mechanism, and potential carcinogenicity of CUL2 itself is still unknown as yet. Here, we demonstrated that CUL2 was specifically overexpressed in HPV16 positive cervical cancer cells and tissues, and CUL2 expression was significantly increased along with the cervical lesion progression and positively correlated with HPV16 E7. CUL2 knockdown slowed the growth of xenograft tumors in mouse models. Importantly, CUL2 specifically bound to HPV16 E7, but not HPV18 E7. Moreover, CUL2 acted as a direct target of miR-424, and reversely suppressed miR-424; E2F transcription factor 1 (E2F1) suppressed miR-424 expression; CUL2 bound to E2F1 and promoted E2F1 expression. Our results indicate the existence of a regulatory loop among CUL2, E2F1, and miR-424 in HPV16 positive cervical cancer cells. Our results suggest that E7 recruited CUL2, driven by CUL2/E2F1/miR-424 regulatory loop, is overexpressed and accelerates HPV16-induced cervical carcinogenesis. Our findings may serve as one of the explanations for a clinical phenomenon that HPV16 possesses the strongest cervical carcinogenicity among high-risk HPV genotypes.

MicroRNA detection in cervical exfoliated cells as a triage for human papillomavirus-positive women.

BACKGROUND: Papanicolaou (Pap) triage, with high specificity, has been recommended for primary Human papillomavirus (HPV) testing but is flawed by poor sensitivity and cytologist dependence. We evaluated the potential role of microRNA (miRNA) detection in cervical exfoliated cells in HPV-positive women from a clinic-based population. METHODS: Primary HPV testing as well as Pap test were performed on all eligible women. Six miRNAs (miR-424/miR-375/miR-34a/miR-218/miR-92a/miR-93) were detected by RT-qPCR in cervical exfoliated cells. All HPV-positive women underwent colposcopy and further biopsy if indicated. Mann-Whitney U test, the receiver operating characteristic curve, logistic regression, and Pearson's Chi-square were used to assess data. All tests of statistical significance were two-sided. RESULTS: A total of 1021 eligible HPV-positive women were enrolled. The expression of miR-424/miR-375/miR-34a/miR-218 in high-grade cervical intraepithelial neoplasia (CIN) and abnormal cytology was statistically significantly lower than that in low-grade CIN and normal cytology, respectively (all P < .05). Compared with the Pap test, both miR-424 and miR-375 detection achieved higher sensitivity (76.0% and 74.9% vs 63.8%, P < .05), higher negative predictive value (NPV) (85.7% and 85.4% vs 79.3%, P < .05), and comparable specificity while identifying CIN2 or worse (CIN2+). Similar results were achieved while identifying CIN3+. Multi-marker panels based on miR-424, miR-375, and miR-218 further improved the performance over any single miRNA test or Pap test. CONCLUSION: Single miR-424 or miR-375 detection and miR-424/miR-375/miR-218-based multimarker panels in cervical exfoliated cells show superior performance over Pap triage for high-grade CIN identification in a clinic-based population. Detection of miRNA may provide a new triage option for HPV-positive women.