Expanded Histopathology and Tropism of Ebola Virus in the Rhesus Macaque Model: Potential for Sexual Transmission, Altered Adrenomedullary Hormone Production, and Early Viral Replication in Liver.

The pathogenesis of Ebola virus disease (EVD) is still incomplete, in spite of the availability of a nonhuman primate modelfor more than 4 decades. To further investigate EVD pathogenesis, a natural history study was conducted using 27 Chinese-origin rhesus macaques. Of these, 24 macaques were exposed intramuscularly to Kikwit Ebola virus and euthanized at predetermined time points or when end-stage clinical disease criteria were met, and 3 sham-exposed macaques were euthanized on study day 0. This study showed for the first time that Ebola virus causes uterine cervicitis, vaginitis, posthitis, and medullary adrenalitis. Not only was Ebola virus detected in the interstitial stromal cells of the genital tract, but it was also present in the epididymal and seminal vesicular tubular epithelial cells, ectocervical and vaginal squamous epithelial cells, and seminal fluid. Furthermore, as early as day 3 after exposure, Ebola virus replicative intermediate RNA was detected in Kupffer cells and hepatocytes. These findings in the nonhuman model provide additional insight into potential sexual transmission, possible disruption of sympathetic hormone production, and early virus replication sites in human EVD patients.

Prevalence and genotype distribution of high-risk HPV infection among women in Beijing, China.

Cervical cancer (CC) is highly associated with high-risk human papillomavirus (HPV) infection and genotype distribution of high-risk HPV (HR-HPV) infection varies greatly in different regions. Clinical specimens were collected from 46 365 patients at Beijing Friendship Hospital, Capital Medical University from January 2017 to December 2020. HPV DNA genotype testing was performed using real-time PCR. The infection rates based on disease group were compared using the χ 2 test. The linear-by-linear association test and gamma value were used to assess the changes in HPV prevalence over calendar year and age group. A total of 10 514 women were infected with HR-HPV, with an overall positive rate of 22.7%. The most prevalent HR-HPV types were HPV52, 58, 16, 51, and 66, and HPV59 had a higher prevalence except for HPV16, 58, and 52 in the CC group. Single infection of HR-HPV was dominant among different disease groups. The infection rate of HR-HPV decreased first and then increased from below 20 years old to over 60 years old. There were significant differences in the HR-HPV infection rates among the age and disease groups. Our findings demonstrate that the genotype distribution of HR-HPV varied with age and diseases. The HR-HPV genotypes prevalence was found to be directly useful for local governments to promote HPV targeted vaccination in the study region.

Clinical Value of Human Papillomavirus E6/E7 mRNA Detection in Screening for Cervical Cancer in Women Positive for Human Papillomavirus DNA or.

BACKGROUND: The detection of human papillomavirus (HPV) E6/E7 mRNA indicates a risk of further deterioration in cervical lesions. We explored the clinical value of HPV E6/E7 mRNA detection in cervical cancer screening in women positive for HPV or with abnormal thin-prep cytology test (TCT) results in the Xinjiang region of China. METHODS: A total of 6,800 women were screened in our hospital for cervical cancer by both TCT and HPV DNA testing from August 2013 to June 2015. Of these, 197 had abnormal cytological or HPV test results and subsequently underwent HPV E6/E7 mRNA detection and histopathological examination, while 101 underwent an HPV DNA typing test. Using pathological results as the gold standard, we compared the accuracies of HPV E6/E7 mRNA detection or HPV DNA type testing alone, in parallel, and in series for diagnosing high-grade cervical lesions. RESULTS: Pathological examination revealed 80 cases of chronic cervicitis, 16 cases of cervical intraepithelial neoplasia (CIN)-I, 50 cases of CIN-II-III, and 51 cases of cervical cancer. The area under the receiver operating characteristic (ROC) curve (AUC) for diagnosing high-grade cervical lesions by HPV E6/E7 mRNA detection was 74.95% (sensitivity, 85.15%; specificity, 66.67%; Youden index, 0.139; positive predictive value, 72.9%; negative predictive value, 81.0%; positive likelihood ratio, 2.555; negative likelihood ratio, 0.222; and post-test probability, 72.9%). CONCLUSIONS: HPV E6/E7 mRNA detection is superior to HPV DNA type testing for diagnosing high-grade cervical lesions.

Human Papillomavirus Genotyping by Surface Plasmon Resonance-Based Test.

BACKGROUND: This study was performed to evaluate the surface plasmon resonance-based (SPR) test with the W2600 System of GP Medical Technologies as a screening method for human papillomavirus (HPV) genotyping in China. METHODS: Between November 2012 and December 2013, the SPR test, hybrid capture II (HC2) test, and direct DNA sequencing assay were performed on 424 patients aged 22 - 65 years who attended the outpatient clinic at the Department of Obstetrics and Gynecology of West China Second University Hospital with diagnoses of chronic cervicitis or abnormal vaginal bleeding. Cervical specimens were collected from the cervical epithelia. Cytological and histological studies were also performed. RESULTS: A total of 424 patient specimens were screened. The positive, negative, and overall concordance between the SPR test and HC2 test was 93.48%, 89.16%, and 90.57%, respectively. The concordance between the SPR test and the direct DNA sequencing assay was 97.41%. Against the results of the direct DNA sequencing assay, the SPR test demonstrated a sensitivity of 96.86% and a specificity of 97.74%. CONCLUSIONS: The SPR test is reliable, sensitive, and specific for HPV genotyping. Compared to the hybrid-capture assay, the SPR test may work on more than one specimen at a time and is a low-cost, rapid, and easy-to-use method with the potential for automation. The SPR test may be widely employed in China and other countries for HPV genotyping.

[Low rate of oropharyngeal human papillomavirus infection among women with cervical lesion. Preliminary results from the South-Eastern Hungarian population]. / Oropharyngealis humán papillomavírus ritka elofordulása cervicalis laesióval rendelkezo nokben. Elozetes eredmények dél-magyarországi vizsgálat alapján.

INTRODUCTION: Although the natural history of cervical and oral human papillomavirus infection has been intensively investigated in the past years, the ability of this virus to infect oral and genital mucosae in the same individual and its potential to co-infect both cervical and oral mucosa are still unclear. AIM: The aim of the authors was to assess the presence of oropharyngeal human papillomavirus infection in women with cervical lesions in the South-Eastern Hungarian population. METHOD: The total of 103 women have been included in the study between March 1, 2013 and January 1, 2015. Brushing was used to collect cells from the oropharyngeal mucosa. Human papillomavirus DNA was detected using polymerase chain reaction, and Amplicor line blot test was used for genotyping. RESULTS: Oropharyngeal human papillomavirus infection was detected in 2 cases (3%). The detected genotypes were 31, 40/61 and 73 in the oropharyngeal region. CONCLUSIONS: The results indicate that in women with cervical lesions oropharyngeal human papillomavirus infection rarely occurs.

[Relationship of Human Papillomavirus Subtypes and Multiple Infection with Different Cervical Precancerous Diseases in Sichuan Province].

OBJECTIVE: To investigate the relationship of human papillomavirus (HPV) subtypes and multiple infections with different cervical precancerous diseases. METHODS: Retrospective study was done to review 1 226 patients with different cervical lesions who were pathologically diagnosed and scanned for HPV 23 subtypes with positive results from June 2006 to May 2012. These patients were divided into the following groups, chronic cervicitis, cervical condyloma, cervical intraepithelium neoplasia grade I (CIN I), grade II (CIN II), grade III (CINIII). RESULTS: There were significant differences in the proportion of HPV low risk types and high risk types between cervicitis, condyloma, CIN I group and CIN II + III groups (P<0. 05). HPV low risk types in condyloma group were mainly 6 and 11 subtype, while the other four groups were 42 and 43 subtype. The four most prevalence high risk types were 58, 16, 52,18 subtype. The infection rates of HPV16 were significant different in cervicitis (11. 0%), CIN II (20. 3%), and CIN III (20. 2%)(P<0. 01), and the infection rates of HPV58 was quite different between cervicitis (15. 9%) and CIN II (21. 4%) (P<0. 05). HPV multiple infection rate in condyloma (68. 8%) was significant different from that of cervicitis (23. 1%), CINI (26. 1%), CIN II (27. 8%) and CIN III (27. 1%) (P<0. 01); while the rest four groups were not significantly different (P>0. 05). CONCLUSION: There is a unique epidemiologic characteristic of HPV infection in Sichuan Province. The HPV low risk types were mainly 42 and 43, and high risk types were mainly 58, 16, 52, 18. It seems that HPV multiple infection is not the leading cause of progression of cervical disease.

High-risk human papilloma virus management in pregnancy with cervical intraepithelial neoplasia during pregnancy and postpartum in China.

AIM: To investigate the relationship between cervical intraepithelial neoplasia (CIN) and high-risk human papilloma virus (HR-HPV) during pregnancy and postpartum in China. METHODS: In this prospective case-control study, 168 pregnant women with CIN and cervicitis were diagnosed by colposcopic cervical biopsy. All the cases underwent hybrid capture assay version II (HCII) to detect HR-HPV DNA load amounts and the tests were completed in 3-6 months after childbirth. RESULTS: During pregnancy: as the CIN grade increased, the HR-HPV infection rates increased (P = 0.002), but HR-HPV DNA load amounts (in logarithms) did not change obviously (P = 0.719). 3-6 months postpartum: as the CIN grade increased, the natural negative rate of HR-HPV decreased (P = 0.000), while the amount of HR-HPV DNA (in logarithms) increased (P = 0.036); especially the amount of HR-HPV DNA in pregnant women with CINIII was significantly higher than that of other grades. During pregnancy and 3-6 months postpartum : the amount of HR-HPV DNA (in logarithms) during pregnancy was higher than that of 3-6 months postpartum with the same grade of CIN. CONCLUSION: The findings emphasize the importance of undergoing the HCII test 3-6 months postpartum. It should be noted that HR-HPV may turn negative in pregnancy with CINIII 3-6 months after childbirth. Further treatments of pregnancy with CIN should be considered according to the CIN grade diagnosed by cervical biopsy via colposcopy 3-6 months after birth, but not according to the persistence of HR-HPV during pregnancy.

Atypical presentation of herpes simplex virus 2 primary infection: a case report.

BACKGROUND: Cervicitis, an infectious or noninfectious inflammation of the cervix, encompasses a wide range of clinical conditions, from asymptomatic infections to severe lesions, making its diagnosis difficult. Acute cervicitis may develop into pelvic inflammatory disease. In patients with cervicitis, current guidelines recommend testing for herpes simplex virus when external genital lesions are present. Here, we present the case of a patient with an atypical primary herpes simplex virus 2 infection manifesting as cervicitis without genital lesions. CASE PRESENTATION: A 29-year-old Caucasian woman was hospitalized for pelvic inflammatory disease. The patient complained of severe suprapubic pain, fever, and heavy vaginal discharge. The external genitalia were unremarkable, so empirical antibiotic treatment was initiated. Despite 48 hours of well-administered antibiotic therapy, her complaints persisted. Polymerase chain reaction for possible microbial causes was negative for Chlamydia trachomatis and Neisseria gonorrhoeae. There was no bacterial vaginosis. Repeat gynecological examinations with endovaginal ultrasound revealed an enlarged cervix, and pelvic magnetic resonance imaging supported a diagnosis of cervicitis. At this point, additional screening for other sexually transmitted infections and infectious disease-related etiologies of cervicitis was performed, and the polymerase chain reaction analysis of newly isolated samples was positive for herpes simplex virus 2. No antiviral treatment was initiated given the delay in diagnosing herpes simplex virus 2 infection and the slow but spontaneous abatement of symptoms. CONCLUSION: Herpes simplex virus infection should be considered as a possible cause of cervicitis, even in the absence of typical genital lesions. Early detection of herpes simplex virus allows early treatment, helping to reduce the duration and severity of symptoms and therefore potentially reducing recurrences and improving disease control. These data and data from future cases might spur changes in the guidelines on cervicitis testing and treatment.

Torque teno mini virus infection in chronic cervicitis and cervical tumors in Isfahan, Iran.

OBJECTIVES: Torque teno mini virus (TTMV) is classified as the Betatorquevirus genus of Anelloviridae. Little is known about the prevalence of TTMV in humans. Worldwide, cervical cancer is the second most common cancer affecting women. This study aimed to estimate the TTMV infection frequency in cervicitis cases and cervical tumors including intraepithelial neoplasia (CIN), squamous cell carcinoma (SCC) and adenocarcinoma, and the possible role of this virus in the etiology of them in an Isfahan population. METHODS: 79 cervicitis cases and 42 tumors were collected from histopathological files of Al-Zahra Hospital in Isfahan, Iran. DNA was extracted and subjected to nested polymerase chain reaction. RESULTS: Totally 62% of the tested samples were positive for TTMV. It was positive in 52.4% of adenocarcinoma, 68.4% of CIN and 100% SCC cases. In cervicitis, 48% of the cases were positive. In the phylogenetic construct two of the cervical tumor isolates and two of the cervicitis isolates were placed in the same cluster with already reported isolates from Japan (EF538880 and AB041962). Also, three of the cervical tumors isolated (JQ734980, JQ734981 and JQ734982) were placed in another cluster. CONCLUSION: The presence of the virus in cervical tissues suggests possible sexual transmission of the virus.

Prevalence and predictors of cervicitis in female sex workers in Peru: an observational study.

BACKGROUND: Cervicitis is a syndrome of cervical inflammation and a common condition in female sex workers (FSW), a subpopulation vulnerable to sexually transmitted infections. Local data is essential for guiding syndromic management of cervicitis in FSW working in Peru. We sought to describe the prevalence and etiologies of cervicitis in this population. We also aimed to identify sociodemographic, behavioral and biological factors associated with cervicitis, including bacterial vaginosis (BV), a condition with a possible role in cervicitis. METHODS: FSW 18 years of age or older presenting to a free public sexual health clinic in Callao-Lima, Peru were eligible for inclusion upon consent. 467 participants completed a face-to-face questionnaire and underwent genital examination. Vaginal, endocervical and blood samples were collected and tested for C. trachomatis (CT), N. gonorrhea (GC), T. vaginalis (TV), BV, HIV and Human T-Cell Lymphotropic Virus -1. Logistic regression was used to determine whether sociodemographic, behavioral, or other sexual health related characteristics were associated with the diagnosis of cervicitis. RESULTS: Cervicitis was detected in 99 (24.9%) of 397 FSW. The presence of cervicitis was unable to be determined in 70 participants. In women with cervicitis, CT was present in 4.6% (4/87), TV in 4.0% (4/99), GC in 0% (0/87) and no pathogen was detected on cervical microbiology in 91.9% (91/99). BV was detected on vaginal microbiology in 36.9% (31/84) of cervicitis cases. BV was more common in women with cervicitis, however this association did not reach statistical significance (aOR = 1.47 [0.87, 2.48], p = 0.15). Other STI were not associated with cervicitis. Regular clinic attendance (aOR = 0.54 [0.34, 0.87], p = 0.01) and Ecuadorian nationality (aOR = 0.31 [0.13, 0.76], p = 0.01) were associated with reduced risk of cervicitis. CONCLUSIONS: Cervicitis was common in FSW working Peru and was predominantly nongonococcal and non-chlamydial in etiology. Further study is warranted to clarify the role of BV and other emerging cervicitis pathogens in this population. The current Peruvian program of free health checks for FSW may be effective for reducing rates of cervicitis. The protective effect of Ecuadorian nationality prompts further study.

Human papillomavirus reactivation following treatment of genital graft-versus-host disease.

Vaginal chronic graft-versus-host disease (cGVHD) is a common complication of stem cell transplantation. Human papillomavirus (HPV) disease can reactivate after transplantation, presumably because of immune factors affecting systemic immunity, such as waning antibody titers, impaired T- and B-lymphocyte responses, and the use of immunosuppressive therapies. However, a relationship between the use of local immunosuppressive agents and HPV reactivation and spread has not been previously described, to our knowledge. A 30-year-old woman, 2 years post transplant receiving systemic cyclosporine for cGVHD, was treated with vaginal dilators, topical corticosteroids, and estrogen for vaginal cGVHD. Colposcopy and biopsy for abnormal cytology revealed condylomatous cervicitis. Over the next 4 months, while continuing dilator therapy, linear verrucous lesions developed in the vagina and vulva, and were successfully treated with laser therapy. Use of local immunosuppression and dilators for genital GVHD can enhance spread of HPV infection. Integration of HPV screening and treatment into the care of women with genital cGVHD and development of strategies to manage both conditions simultaneously are warranted.

Genital tract inflammation during early HIV-1 infection predicts higher plasma viral load set point in women.

BACKGROUND: The biggest challenge in human immunodeficiency virus type 1 (HIV-1) prevention in Africa is the high HIV-1 burden in young women. In macaques, proinflammatory cytokine production in the genital tract is necessary for target cell recruitment and establishment of simian immunodeficiency virus (SIV) infection following vaginal inoculation. The purpose of this study was to assess if genital inflammation during early HIV-1 infection predisposes women to rapid disease progression. METHODS: Inflammatory cytokine concentrations were measured in cervicovaginal lavage (CVL) from 49 women 6, 17, 30, and 55 weeks after HIV-1 infection and from 22 of these women before infection. Associations between genital inflammation and viral load set point and blood CD4 cell counts 12 months after infection were investigated. RESULTS: Elevated genital cytokine concentrations 6 and 17 weeks after HIV-1 infection were associated with higher viral load set points and, to a lesser extent, with CD4 depletion. CVL cytokine concentrations during early infection did not differ relative to preinfection but were elevated in women who had vaginal discharge, detectable HIV-1 RNA in their genital tracts, and lower blood CD4 counts. CONCLUSION: Genital inflammation during early HIV-1 infection was associated with higher viral load set point and CD4 depletion, which are markers of rapid disease progression. Strategies aimed at reducing genital inflammation during early HIV-1 infection may slow disease progression.

[The use of human papillomavirus genotying kit based on gene chip technology for detecting and typing of human papillomavirus].

OBJECTIVE: To evaluate a new human papillomavirus (HPV) genotyping technique based on gene chip technology (HPG) for HPV genotyping and its clinical efficacy. METHODS: HPV genotyping (HPG) test, hybrid capture II (HC2) test and DNA sequencing assay were performed in 151 patients aged 20-75 years with diagnosis of chronic cervicitis or abnormal vaginal bleeding. The cervical specimens were collected from cervical epithelium. All the cervical samples were analyzed by the HPG test, HC2 test and DNA sequencing. The clinical efficacy of the HPG test was analyzed. RESULTS: The consistent rate between HPG test and HC2 test was 87.42% (kappa = 0.75, P < 0.05). When DNA sequencing assay was regarding as the final test result, the sensitivity and specificity of HPG test for high risk HPV were 100% and 96.49%, respectively. The consistent rate between HPG test and direct DNA sequencing was 98.70% (kappa = 0.97, P < 0.05). The most common six HPV genotypes detected by HPG test were HPV 16 (13.25%), 58 (11.92%), 52 (11.92%), 31 (6.62%) 39 (5.96%), 33 (5.96%) in descending order of frequency. The incidence of multiple-types infection detected by HPG test was 23.84%. CONCLUSION: HPG test is a rapid and accurate test for HPV genotyping which could detect 29 types of HPV infection at one time. It is suitable for cervical HPV infection screening in clinic.

Relationship between cervical disease and infection with human papillomavirus types 16 and 18, and herpes simplex virus 1 and 2.

Persistent infection with high-risk HPV, particularly Type HPV 16 and 18, is necessary in the development of cervical cancer, but apart from HPV infection, other causative factors of most cervical cancers remain unknown. The aim of this study was to determine the prevalence of HPV 16 and HPV 18 and HSV 1 and HSV 2 in cervical samples, and to assess the role of HSVs in cervical carcinogenesis. Two hundred thirty-three healthy controls and 567 cases (333 of cervicitis, 210 of cervical intraepithelial neoplasia, and 24 of squamous cell carcinoma) in cervical exfoliative cells were tested for HPV 16, HPV 18, HSV 1, and HSV 2 DNA using the triplex real-time polymerase chain reaction method. In contrast to healthy women, positive rate of HPV is related significantly to cervical lesions (odds ratios (ORs) = 4.1, P < 0.01 for cervical intraepithelial neoplasia; ORs = 24.9, P < 0.01 for squamous cell carcinoma), but not cervicitis (ORs = 2.3, P > 0.05). HSV 2 prevalence in cervical intraepithelial neoplasia and squamous cell carcinoma was higher than in healthy women (ORs = 4.9, P < 0.05 for cervical intraepithelial neoplasia; ORs = 4.7, P < 0.05 for squamous cell carcinoma). HSV 2 coinfection with HPV in cervical intraepithelial neoplasia and squamous cell carcinoma was strongly higher than in healthy women (ORs = 34.2, P < 0.01 for cervical intraepithelial neoplasia; ORs = 61.1, P < 0.01 for squamous cell carcinoma). The obtained results indicated that the presence of HPV is associated closely with cervical cancer, and that HSV 2 infection or co-infection with HPV might be involved in cervical cancer development, while HSV 1 might not be involved.

Detection of torque teno midi virus/small anellovirus (TTMDV/SAV) in chronic cervicitis and cervical tumors in Isfahan, Iran.

Torque teno midi virus and small anellovirus (TTMDV/SAV) are members of the genus Gammatorquevirus within the family Anelloviridae. Cervical cancer is the second most prevalent cancer after breast cancer. The aim of this study was to determine the frequency of infection by these viruses in cervicitis and cervical tumors of women from Isfahan, Iran. Formalin-fixed, paraffin-embedded tissue samples from cervical cancers (n = 42) and cervicitis cases (n = 79) were subjected to nested PCR to identify TTMDV/SAV viral sequences. Of the 42 tumor cases, 22, 18 and 2 were diagnosed as adenocarcinoma, cervical intraepithelial neoplasia and squamous cell carcinoma, respectively. In total, 23 (55%) of the tumor samples were positive for TTMDV/SAV. Of the 79 cervicitis cases, 38 (48%) were also positive for TTMDV/SAV. This is the first report of TTMDV/SAV in cervicitis and cervical tumors of women.

Expression of P-Akt, NFkappaB and their correlation with human papillomavirus infection in cervical carcinoma.

PURPOSE: To investigate the expression of P-Akt and NFkappaB and their correlation with human papillomavirus (HPV) infection in cervical carcinoma. MATERIAL AND METHODS: Expression of P-Akt and NFkappaB was detected by an immunohistochemical SP technique with HPV DNA detetion by PCR in 26 cases of cervical carcinoma tissues, 18 cases of cervical intraepithelial neoplasia tissues (CINI / n = 5, CINII / n = 3, CINIII / n = 10) and 19 cases of chronic cervicitis tissues. The different expressions of P-Akt and NFkappaB were compared in different pathological types of cervical carcinoma (cervical squamous cell carcinoma, cervical adenocarcinoma), different pathological grading (high, medium, poorly differentiated) and different clinical stage (FIGO I to IV). The relationships between P-Akt and NFkappaB, respectively, with HPV infection in cervical carcinoma were analyzed. RESULTS: The positive expression rate of P-Akt in chronic cervicitis tissues, CIN and cervical carcinoma tissues was 21.05%, 66.67%, and 92.31%, respectively. There was no obvious difference in the expression of P-Akt in cervical carcinoma in different pathological types or in pathological grading and no obvious difference in different clinical stages. The positive expression rate of NFkappaB in chronic cervicitis tissues, CIN and cervical carcinoma tissues was 10.52%, 72.22% and 96.15%, respectively; there was no statistically significant difference among the groups for different pathological types and there was no obvious difference in different pathological grading or different clinical stage. There was an obviously positive correlation between P-Akt and NFkappaB expression rate and degree of disease (r = 0.998, p < 0.05). Cervical carcinoma and CIN cases totaled 44; the positive expression rate of P-Akt was 87.55% in 32 cases of positive HPV-DNA of the 44 cases, and the positive expression rate of P-Akt was only 16.70% in 12 cases of negative HPV-DNA of the 44 cases. The positive expression rate of NFkappaB was obviously higher in the HPV DNA positive than in the HPV-DNA negative cases. There was a statistically significant difference among the groups (p < 0.05). CONCLUSION: The positive expression rate of P-Akt and NFkappaB was closely related with cervical disease extent, and closely related with HPV infection in cervical carcinoma. This study suggests that P-Akt and NFkappaB more probably play an important role in the occurrence of cervical carcinoma.

HPV detection methods and genotyping techniques in screening for cervical cancer.

Human papillomaviruses (HPV), double-stranded DNA viruses, are causing many mucocutaneous diseases, benign or malignant, ranging from common warts to malignancies involving the upper aerodigestive tract and the anogenital sphere. The diagnosis of HPV infection is based primarily on the viral genome detection by molecular biological methods, given the difficulty in routine cultivation of these viruses. The current trend in screening against cervical cancer is to improve the sensitivity of screening with new methods and to propose new algorithms for diagnostic and therapeutic decisions. The development of liquid-based cytology facilitates the cytologic diagnosis and molecular assays from the same sample. There are two main types of HPV detection methods used on uterine cervical samples: signal amplification methods (hybridization techniques in liquid phase) and target amplification methods (the techniques of gene amplification or Polymerase Chain Reaction [PCR]). Genotyping techniques are also developed: they are based on an amplification technique followed by hybridization with probe specific types. In addition to the detection, genotyping techniques allow quantitative detection of viral DNA of HPV genotype and so monitor changes in viral load over time. Another approach relies on the detection of messenger RNA (mRNA) of HPV proteins E6 and E7 oncogenes, which would appear to be a relevant marker to identify and monitor women at risk of progression to a precancerous lesion or cervical cancer.

[Clinical study on cervical intraepithelial neoplasia with high-risk HPV infection among pregnant women].

OBJECTIVE: To investigate the relationship between cervical intraepithelial neoplasia (CIN) and high-risk (HR) HPV infection among late pregnant women. METHODS: From Aug. 2007 to Feb. 2010, 168 women at 13 to 32 gestational weeks undergoing prenatal examination in Beijing Obstetrics and Gynecology Hospital went through three stage cervical disease screening, including 21 women with cervicitis and 147 women with CIN (42 women with CIN III, 37 women with CIN II and 68 women with CIN I). Hybrid capture assay version II (HC-II) test was used to measure HR-HPV DNA load, and the logarithmic transtormation (log(10)) was performed. All 168 women were followed up to postpartum 3 - 6 months. HR-HPV infections rates of cervicitis and different CIN, the rate of HR-HPV infection turned naturally negative at postpartum of 3 to 6 months, and HR-HPV load at pregnancy and 3 - 6 months postpartum were observed. RESULTS: (1) HR-HPV infection rate: CIN III, II, I and cervicitis pregnant women's HR-HPV positive infection rates were 98% (41/42), 86% (32/37), 76% (52/68) and 62% (13/21) respectively, which reached statistical difference (P = 0.002). (2) HR-HPV naturally negative: the rate of pregnant women with different levels of CIN who turned HR-HPV naturally negative within 3 - 6 months of postpartum were CIN III 5% (2/41), CINII 47% (15/32), CINI52% (27/52) and cervicitis 10/13, which also reached statistical difference among those four groups (P = 0.000). (3) HR-HPV load: pregnant women with different grade of CIN and cervicitis HR-HPV DNA load were CIN III 2.02 ng/L(1.53, 2.67 ng/L), CINII 1.94 ng/L (0.75, 2.75 ng/L), CINI2.04 ng/L (0.08, 2.95 ng/L) and cervicitis 1.98 ng/L (-0.07, 2.47 ng/L). There was no significantly different HPV load in women with cervicitis and different CIN (P = 0.719). At 3 - 6 months postpartum, HR-HPV load was CIN III 1.55 ng/L (0.90, 2.10 ng/L), which was significantly higher than the amount of CINII 0.09 ng/L (-0.69, 1.74 ng/L), CINI0.48 ng/L (-0.56, 2.2 ng/L) and cervicitis -0.46 ng/L (-0.78, 1.40 ng/L, P = 0.036). CONCLUSIONS: With the increasing of CIN grade, the rate of HR-HPV infection in pregnant women was increased, however, the rate of HR-HPV turning negative naturally at 3 - 6 months postpartum decreased. With different CIN grade during pregnancy, HR-HPV DNA load did not change significantly, but HR-HPV DNA load increased at 3 - 6 months of postpartum. HR-HPV DNA loads with the same grade of CIN and cervicitis during pregnancy higher than that of postpartum among pregnant women.

[Significance of DNA ploidy analysis in diagnosis of ASCUS].

OBJECTIVE: To investigate the significance of DNA ploidy analysis in diagnosis of atypical squamous cell of undetermined significance (ASCUS). METHODS: From Jan.2009 to Jul.2011, 875 women with ASCUS confirmed by liquid based thin layer cytology technique underwent DNA ploid analysis in Hubei Maternal and Child Health Hospital. Among 294 women underwent high risk HPV detection. All subjective were examined colposcopy directed biopsy at day 3 to 10 after menstruation. RESULTS: Among 875 ASCUS cases, 553 cases with histologically as chronic cervicitis (63.2%), 165 cases with cervical intraepithelial neoplasia (CIN)I (18.9%), 45 cases with CINII (5.1%), 79 cases with CINIII (9.0%) and 33 cases with cervical invasive cancer (3.8%) were confirmed by colposcopy. Totally 532 cases were observed with DNA heteroploid, and 343 were not observed with DNA heteroploid. When DNA heteroploid negative and more than or equal to three ploid were used to predict CINII or more severe cervical diseases, the sensitivity, specificity, positive predictive values and negative predictive values were 98.7% and 90.3%, 47.5% and 46.1%, 29.1% and 40.8%, 99.4% and 92.1%, respectively. The amount of heteroploid cells > 2.5c and > 5c among every 100 detected cells in chronic cervicitis and CINI, CINII, CINIII and cervix cancer were respectively 2.53 ± 1.99 and 0.10 ± 0.07, 2.24 ± 1.69 and 0.20 ± 0.11, 4.10 ± 1.91 and 0.28 ± 0.19, 7.97 ± 7.33 and 1.27 ± 1.23, 8.99 ± 7.33 and 0.36 ± 0.33, there was no statistical difference in amount of heteropolid cells between >2.5c and > 5c at group of chronic cervicitis and CINI, CINIII and cervix cancer (P > 0.05). However, the amount of heteroploid cells at > 2.5c and > 5c at group of chronic cervicitis, CINI, CINIII and cervical were higher than that of CINII significantly (P < 0.05). Among 294 cases with high risk (HR) HPV detection, 216 cases were HR-HPV positive, and 78 cases were HR-HPV negative. The pathology result by colposcopy at group of negative heteroploid, heteroploid < 3, or ≥ 3 showed statistical distribution (χ(2) = 115.2775, P < 0.01). CONCLUSION: DNA ploidy analysis can be used for ASCUS diagnosis, which can avoid excessive biopsy under colposcopy, in the mean time, CIN and cervical cancer could decrease missed diagnosis.