高危型人乳头瘤病毒阴性/不典型鳞状细胞和低度鳞状上皮内病变患者的随访观察

目的分析高危型人乳头瘤病毒(high-risk human papillomavirus,HR-HPV)阴性的不典型鳞状细胞(atypical squamous cells undetermined significance,ASCUS)和低度鳞状上皮内病变(low squamous intraepithelial lesion,LSIL)患者的随访结果,探讨HR-HPV阴性的ASCUS和LSIL患者的合理处理方法。方法回顾性分析2010年1月1日至2012年12月31日期间,就诊于首都医科大学附属北京朝阳医院妇科门诊,HR-HPV阴性、宫颈液基细胞学为ASCUS和LSIL的356例患者的病理资料,对HR-HPV阴性的ASCUS和LSIL经阴道镜及宫颈活检排除高级别病变的306例患者定期进行HR-HPV随访。HR-HPV检测采用杂交检测法2代(hybird capture-Ⅱ,HC-Ⅱ)。结果 HR-HPV阴性/ASCUS和LSIL患者宫颈上皮内瘤变(cervical intraepithelial lesions,CIN)-Ⅲ及以上病变发病率分别为7.9%和9.1%。HR-HPV阳性/ASCUS和LSIL患者CIN-Ⅲ及以上病变发病率分别是22.9%和17.9%。HRHPV阳性的ASCUS和LSIL患者CIN-Ⅲ及以上病变发病率明显高于HR-HPV阴性患者,差异具有统计学意义(P=0.000)。HRHPV阳性对ASCUS及LSIL患者宫颈高级别病变的阴性预测值分别为87.13%和84.42%。HR-HPV阴性/ASCUS和LSIL患者CIN-Ⅱ及以上病变检出率是14.3%。不同年龄段的HR-HPV阴性/ASCUS和LSIL的患者,宫颈高级别病变患者的发生率不同,差异具有统计学意义(P<0.05)。30~39岁妇女宫颈高级别病变的发生率最高(18.2%)。结论 HR-HPV阴性的ASCUS和LSIL患者发生CIN-Ⅲ及以上病变分别是7.9%和9.1%,CIN-Ⅱ及以上病变达14.3%,30~39岁的HR-HPV阴性/ASCUS和LSIL患者宫颈高级别病变发生率达18%,建议对于HR-HPV阴性的ASCUS和LSIL患者应行阴道镜检查可疑病变部位活检。     

Cervical Infection of Oncogenic Human Papillomavirus (HPV) Types in Beijing,China

ObjectiveThis study was designed to determine the prevalence of oncogenic human papillomavirus (HPV) in cervical infections in Beijing, China, and to investigate the odds ratio (OR) of HPV single and multiple infections in abnormal cytology. MethodsA total of 19,018 specimens from outpatients in thedepartment ofobstetric andgynecology were collected. They were detected usinghigh-risk HPV genotyping real-time polymerase chain reaction (PCR) kit and analyzed by ThinPrep cytology test for cervical pathological diagnosis. HPV prevalence, age-specific prevalence, andOR of each type of HPV in abnormal cytology were analyzed. ResultsOverall, 19.1% (3,623/19,018) of the individuals were positive for HPV infection, 14.9%(2,833/19,018) were positive for a single HPV type, and 4.2% (790/19,018) were positive for multiple types. Among the 3,623 HPV-positive individuals, the most predominant HPV types were HPV52 (4.4%, 834/19,018), HPV16 (3.7%, 710/19,018), and HPV58 (3.4%, 644/19,018). TheOR of multiple infections and single infection differed significantly among disease severities. TheOR of dual infection was higher than that of each of the two single infection types, respectively. ConclusionHPV prevalence in the outpatients was 19.1%, and the most predominant HPV types in the study were HPV52, HPV16, and HPV58. Women with multiple infectionswere more likely to have abnormal cytology.     

Pap Smear miR-92a-5p and miR-155-5p as potential diagnostic biomarkers of squamous intraepithelial cervical cancer

Background: one of the female-specific diseases with a high incidence and mortality is cervical cancer. The main cause of cervical cancer is infection with Human papilloma virus (HPV). Low-grade squamous intraepithelial lesions (LSIL) and High-grade squamous intraepithelial lesions (HSIL) usually is caused by an HPV infection. Considering the role of microRNAs (miRNAs) as diagnostic biomarkers for a variety of cancers, the aim of this study was to determine miR-92a-5p and miR-155-5p expression levels in LSIL and HSIL Pap Smear samples. Methods: After initial bioinformatic studies, A total of 75 samples (25 samples of patients with LSIL, 25 patients with HSIL and 25 healthy individuals) were subjected to RNA extraction and cDNA synthesis. The expressions levels of confirmed miRNAs in samples of patients with LSIL, HSIL and healthy individuals were evaluated by Real time PCR analysis. To demonstration the role of predicted miRNAs as novel biomarkers in diagnosis of LSIL and HSIL, ROC curve analysis was done. Results: Bioinformatics results showed that miR-92a-5p and miR-155-5p target the HPV E6 and E7 genes. The expression levels of these miRNAs were strikingly higher in Pap smear of patients with LSIL than in the healthy individuals (35.36, P = 0.001) (62.23, P = 0.001). Similarity, expression levels of miR-92a-5p and miR-155-5p were amazingly higher in patients with HSIL than in the healthy individuals (33.62, P= 0.001) (69.07, P= 0.001). Although, the levels of miR-92a-5p (0.95, P = 0. 85) and miR-155-5p (1.11, P = 0.84) exhibited no statistical differences between patients with LSIL and HSIL. Also, ROC curve analyses verified that miR-92a-5p and miR-155-5p are specific and sensitive and may serve as new biomarkers for the early detection of cervical cancer. Conclusion: These data suggest miR-92a-5p and miR-155-5p, which are upregulated in LSIL and HSIL, can be consider as predictive biomarkers for the prognosis of cervical cancer patients.     

Predicting CIN2+ when detecting HPV mRNA and DNA by PreTect HPV-proofer and consensus PCR: A 2-year follow-up of women with ASCUS or LSIL Pap smear

It has been suggested that human papillomavirus (HPV) testing improves follow-up of atypical cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL) in cervical cancer screening programs. To evaluate the prognostic value of including HPV testing as an adjunct to cytology, we carried out a 2-year follow-up study of 77 women with ASCUS or LSIL Papanicolaou (Pap) smear in the Norwegian Cervical Cancer Screening Program (NCCSP) for detection of histological cervical intraepithelial neoplasia (CIN) 2+. The study includes a comparison between viral mRNA and DNA detection. PreTect HPV-Proofer was used for HPV E6/E7 mRNA detection from the 5 high-risk types 16, 18, 31, 33 and 45, and Gp5+/6+ consensus PCR was used for HPV DNA detection. Twice as many women were positive for HPV DNA (54.6%) than for HPV mRNA (23.4%). PreTect HPV-Proofer and consensus PCR had a sensitivity of 85.7% (95% confidence interval [CI] = 42.1-99.6) for detecting CIN2+ during follow-up. The specificity was significantly higher for PreTect HPV-Proofer, 84.9% (95% CI = 73.9-92.5), than for consensus PCR, 50.0% (95% CI = 37.4-62.6). PreTect HPV-Proofer positive women were 69.8 times (95% CI = 4.3-1137.3) more likely to be diagnosed with CIN2+ within 2 years than PreTect HPV-Proofer negative women. Consensus PCR-positive women were 5.7 times (95% CI = 0.6-52.0) more likely to be diagnosed with CIN2+ within 2 years than PCR-negative women. With equal sensitivity and higher specificity than consensus PCR, the PreTect HPV-Proofer might offer an improvement for the triage of women with ASCUS or LSIL Pap smear.     

Human papillomavirus type specific risk of progression and remission during long‐term follow‐up of equivocal and low‐grade HPV‐positive cervical smears

The prevalence of clinically relevant HPV types and their specific risk for progression and regression in women with atypical squamous cells of uncertain significance (ASCUS) and low‐grade squamous intraepithelial lesions (LSIL) were studied in a routine screening population. A 4‐year cohort of women (n = 820) with ASCUS/LSIL and a positive HPV test in triage were followed for 6–9 years. The progression risks for CIN2+/CIN3+ were determined for single (71.2%) and multiple HPV infections (28.8%). The CIN2+ progression risk for all HPV 16, all HPV 35, single HPV 16 and single HPV 35 infections were 65.3% (95% CI: 59.6–71.0), 64.4% (95% CI: 50.4–78.4), 63.8% (95% CI: 56.2–71.4) and 73.7% (95% CI: 53.9–93.5), respectively. Based on CIN2+ progression risks four main groups were defined; the HPV 16 group, the HPV 31/33/35 group, the HPV 18/45/51/52 group and the HPV 39/56/58/59/66/68 group with progression risks of 65.3% (95% CI: 59.6–71.0), 62.1% (95% CI: 54.8–69.4), 52.6 (95% CI: 45.9–59.3) and 39.5 (95% CI: 33.0–46.0), respectively. In multivariate analyses, women in the age group 40–49 years had an increased risk of CIN2+ progression. As for CIN3+, HPV 16 had a higher progression risk than other HPV risk groups (p < 0.05). In multiple infections only HPV 16 had a significant additive CIN3+ progression risk (p < 0.05) as compared to other HPV risk groups. In summary, HPV types 16 and 35, including the HPV risk group 31/33/35, had a similar CIN2+ progression risk, but only HPV 16 had a higher risk for CIN3+ progression.     

Human papillomavirus infection and cervical intraepithelial neoplasia in a cohort of low-risk women

Objective

This study was a prospective examination of the incidence of human papillomavirus (HPV) infection in a low-risk female population and an assessment of the risk of development of LSIL with HPV infection.

Study design

In a longitudinal study, women aged 19–60 years – non-smokers, and married or living with a constant partner, who presented for cervical cancer screening at an outpatient clinic – were invited to participate in a prospective study of cervical HPV infection, and were examined every 3 months.

Results

Of the 464 women at risk, 20 presented with HPV infections during the follow-up. Low-grade squamous intraepithelial lesions (LSIL) event developed in 18 women. Among these women, 13 were HPV-positive (10 high-risk and 3 low-risk types). The average duration of new LSIL was 20.1 months (95% CI: 13.9–26.3) and 55.3 months (95% CI: 45.7–64.9) in the HPV-positive and negative groups, respectively, the difference was statistically significant (p < 0.001).With the use of Cox proportional hazard regression, we estimated the relative risk as 90.0 for a first instance of LSIL among women testing positive for HPV as compared with women testing negative for HPV.

Conclusion

This study has provided evidence that HPV infection is associated with an increased risk of LSIL.     

Flow Cytometric Analysis of DNA Ploidy in Liquid Based Cytology for Cervical Pre-Cancer and Cancer

Background: DNA ploidy analysis of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer samples by flow cytometry (FCM) has been established as an aid to prognostic assessment. Liquid based cytology (LBC) increases diagnostic specificity by using ancillary techniques that provide information beyond morphology. The present study was undertaken to assess DNA ploidy in LBC samples as an adjunct for early detection of cervical pre-cancer and cancer. Methods: DNA ploidy assessment was performed on LBC samples of 50 cases and 31 controls. Cell pellets were obtained by centrifugation and stained with Telford reagent. At least 20,000 R1 gate (G0-G1) events were acquired on a BD FACSCalibur by using a 575±10 nm filter. Results: Mean diploid G1 values were lowered significantly (p<0.01) while diploid S values were significantly elevated (p<0.01) in both high grade squamous intraepithelial lesions (HSILs) and squamous cell carcinomas (SCCs) as compared to controls. Receiver operating curve (ROC) analysis of the diploid G1 value was found to have significant diagnostic potential (AUC=0.682, Z=2.00, p=0.046) for distinction between control and low grade squamous intraepithelial lesion (LSIL) at a cut off value of ≤91.6 with a sensitivity and specificity of 50.0 and 87.1%, respectively. Conclusions: ROC analysis of diploid G1 and diploid S values allows discrimination between LSIL and HSIL with sensitivities and specificities of 65 and 100% and 70 and100%, respectively, and between LSIL and SCC cases with values of 71.4 and 100% and 64.3 and 100%, respectively.     

Evaluation of p16INK4a/Ki‐67 dual stain in comparison with an mRNA human papillomavirus test on liquid‐based cytology samples with low‐grade squamous intraepithelial lesion

BACKGROUND:

The objective of the current study was to investigate the clinical performance of detecting high‐grade lesions with the CINtec PLUS p16^INK4a/Ki‐67 dual stain and the APTIMA human papillomavirus (HPV) Assay in a cohort of women with low‐grade squamous intraepithelial lesion (LSIL) cytology. The authors also assessed the reproducibility of the evaluation of immunocytochemical staining.

METHODS:

The 2 tests were performed on liquid‐based residual material from 469 women with LSILs. The samples had at least 5 years of follow‐up and the gold standard used was high‐grade cervical intraepithelial neoplasia (CIN2+/CIN3+) proven on histology.

RESULTS:

Approximately 69% of all the women included in the study had a positive test for HPV mRNA and 56% was positive for the dual stain. The 2 tests demonstrated high sensitivities. When examining the specificities, the APTIMA HPV Assay performed with significantly lower values than the CINtec PLUS test. For patients with CIN2+, the APTIMA HPV Assay had a specificity of 36.1% versus 51.3% for the CINtec PLUS test, and for women with CIN3+, the specificity was 33.8% versus 48.2%, respectively. The difference was even more pronounced when analyzing women aged < 30 years separately. The kappa values between the 3 observers in scoring the dual stain ranged from 0.43 to 0.49 and improved in a second evaluation round to values ranging from 0.50 to 0.66.

CONCLUSIONS:

The CINtec PLUS p16^INK4a/Ki‐67 dual‐staining test in LSIL cytology samples demonstrated high sensitivity that was similar to that of the APTIMA HPV Assay in the detection of underlying high‐grade disease but with enhanced specificity, especially among women aged < 30 years. The kappa value for the evaluation of the CINtec PLUS dual‐staining test was moderate but could be improved through training. Cancer (Cancer Cytopathol) 2013. © 2012 American Cancer Society.     

Cryotherapy for HPV clearance in women with biopsy-confirmed cervical low-grade squamous intraepithelial lesions

Objective

To compare the clearance rate of HPV infection among women aged older than 30 years with biopsy-confirmed cervical low-grade squamous intraepithelial lesions (LSIL) 1 year after cryotherapy with the spontaneous clearance rate (observation).

Method

HPV DNA typing by polymerase chain reaction and reverse line blot hybridization were used to identify 14 high-risk types and 23 low-risk types. HPV DNA sequencing was also used for other types.

Result

Between December 2007 and March 2009, 100 women were recruited to the study and 60 cases had positive results on HPV testing. Twenty-nine patients were randomly allocated to the cryotherapy group and 31 to the observation group. At 1 year, 89.7% (26/29; 95% CI, 78.6-100%) of the cryotherapy group and 90.3% (28/31; 95% CI, 79.9-100%) of the observation group had negative results on HPV testing (0.6% difference; 95% CI, -15.8 to 14.6%, P = 0.94).

Conclusion

Cryotherapy failed to increase the clearance of prevalent HPV infections among women with LSIL, although in both arms the clearance rates were above 80%. However, in coupling with visual inspection with acetic acid as a single visit approach, its effect on prevention of HSIL and cervical cancer is still promising. Therefore, cryotherapy should not be withdrawn from such programs. (ClinicalTrials.gov Identifier: NCT00566579).