Adverse Pregnancy Outcomes Following Cryotherapy, Thermal Ablation and Loop Electrosurgical Excision Procedure for Cervical Intraepithelial Neoplasia Treatment: A Pilot Study among Zambian Women

Background: Cervical Intraepithelial neoplasia treatments have become essential interventions to manage cervical lesions. Majority of the recipients of these treatments are women within the reproductive age group, who according to literature may be at risk of adverse pregnancy outcomes. This pilot study is part of a study investigating adverse pregnancy outcomes among women who received Cryotherapy, Thermal ablation and Loop Electrosurgical Excision Procedure compared to the untreated women in Zambia. Materials and Methods: This descriptive study analyzed records of 886 (n = 443 treated and n = 443 untreated) women aged 15 - 49 years. The women were either screened with Visual Inspection with Acetic Acid or treated for Cervical Intraepithelial neoplasia at the Adult Infectious Disease Centre between January 2010 and December 2020. Women meeting the criteria were identified using the Visual Inspection with Acetic Acid screening records and telephone interviews to obtain the adverse pregnancy outcome experienced. Data were analysed using STATA version 16 to determine the prevalence and obtain frequency distribution of outcomes of interest. Univariate and multivariable binary logistic regression estimated odds of adverse pregnancy outcomes across the three treatments. Results: The respondents were aged 15 to 49 years. Adverse pregnancy outcomes were observed to be more prevalent in the treatment group (18.5%) compared to the untreated group (5.4%). Normal pregnancy outcomes were lower in the treated (46.3%;n = 443) than the untreated (53.7%;n = 443). The treated group accounted for the majority of abortions (85.2%), prolonged labour (85.7%) and low birth weight (80%), whereas, the untreated accounted for the majority of still births (72.7%). Women treated with cryotherapy (aOR = 2.43, 95% CI = 1.32 - 4.49, p = 0.004), thermal ablation (aOR = 6.37, 95% CI = 0.99 - 41.2, p = 0.052) and Loop Electrosurgical Excision Procedure (aOR = 9.67, 95% CI = 2.17 - 43.1, p = 0.003) had two-, six- and ten-times higher odds of adverse pregnancy outcomes respectively, relative to women who required no treatment. Conclusion: Adverse pregnancy outcomes are prevalent among women who have received treatment in Zambia. The findings indicate that treating Cervical Intraepithelial Neoplasia has been linked to higher chances of experiencing abortion, delivering low birth weight babies and enduring prolonged labor that may result in a caesarean section delivery. Cervical neoplasia treatments, particularly Loop Electrosurgical Excision Procedure, are associated with significantly increased odds of adverse pregnancy outcomes. It is essential to include information about prior Cervical Intraepithelial neoplasia treatment outcomes in obstetric care.

Distribution of High-Risk Human Papillomavirus Genotypes among Women with Colposcopic Diagnosis of Cervical Intraepithelial Neoplasia in Bangladesh

Background: The incidence of cervical cancer is high in Bangladesh and there is a high prevalence of preinvasive lower genital tract disease among women of reproductive age. Persistent high-risk Human Papilloma Virus (HPV) infection is the main underlying cause of cervical cancer and its precursor, cervical intraepithelial neoplasia (CIN). Objective: The aim of the study was to identify the subtypes of high-risk HPV infection among women with the colposcopic diagnosis of cervical intraepithelial neoplasia in Bangladesh. Methods: This cross-sectional observational study was conducted in the colposcopy clinic of Dhaka Medical College Hospital over a six-month period. A total of 100 participants were enrolled. Married women, between 30 - 60 years of age with colposcopically diagnosed cervical intra epithelial neoplasia were enrolled. Women with chronic illness, pregnancy, and women unable to consent were excluded from this study. After counselling, colposcopically directed punch biopsies were taken from each CIN case concurrently with high-risk HPV testing by polymerase chain reaction (PCR). Results: The mean age of the patients was 38.69 (SD ±7.76) years. CIN 1 was diagnosed in 57% of participants, while 24% had CIN II and 19% had CIN III lesions. High-risk HPV was present in 52 patients. HPV 16 was the most common identified in 28 (53.84%) and HPV 18 was the second most common with 20 (38.46%) either singly or in combination with other high-risk subtypes. The other HPV strains, HPV 31, 33, 35, 52, 56 and 58, were also detected either as mono or co-infections. Out of the 52 HPV positive cases, 29 (55.8%) had mono infection and 23 (44.2%) had co-infection with several subtypes. The highest incidence (50%) of oncogenic HPV infections was present among women aged 35 - 45 years. Risk factors associated with HPV positive cases were high parity (P 0.05), early age at marriage (P = 0.754) and early age of first child. Conclusion: This study identified a high prevalence of HPV 16 and 18 genotypes. HPV vaccination with the current 9-valent HPV vaccine, which contains HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58. Will be an effective public health measure to eradicate cervical cancer in Bangladesh.

Prevalence of HPV Infection And Cervical Intraepithelial Neoplasia And Attitudes towards HPV Vaccination among Chinese Women Aged 18-25 in Jiangsu Province

Objective：Few data are available on the epidemiology of HPV and cervical cancer among Chinese women younger than 25 years old.This study aimed to estimate the HPV infection rate and the prevalence of cervical intraepithelial neoplasia（CIN） in women aged 18-25,as well as their knowledge of and attitudes towards HPV vaccination.Methods：A population-based cervical cancer screening study was conducted on women aged 18-25 in Jiangsu province in 2008.Participants provided socio-demographic,reproductive and behavioral information and completed a survey about their knowledge of and attitudes towards HPV vaccination.Women then underwent a gynecologic exam to provide two cervical exfoliated cell samples for high risk HPV DNA testing and liquid-based cytology（LBC） as well as visual inspection with acetic acid（VIA）.Women testing positive for any test were referred to colposcopy and biopsy.The gold standard for diagnosis of cervical lesions was directed or random biopsies.Results：Within the sample of 316 women,3.4% of them were diagnosed with CIN grade 2 or worse lesions and 17.1% were found to be positive for HPV DNA.Among these young women,extra-marital sexual behavior of them（OR=2.0,95%CI：1.1-3.8） or their husbands（OR=2.6,95%：1.4-4.7） were associated with an increased risk of HPV positivity.Although overall HPV awareness was low,after a brief educational intervention,98.4% reported they would electively receive HPV vaccination and would also recommend that their daughters be vaccinated.However,most urban and rural women reported their ideal maximum out-of-pocket contribution for HPV vaccination to be less than 500 RMB and 50-100 RMB,respectively.Conclusion：Our study indicates cervical disease burden is relatively high among sampled Chinese women aged 18-25.Appropriate educational interventions for female adolescents and strategies to subsidize vaccine costs are definitely needed to ensure the effectiveness of vaccination campaigns in China.

Clinical Observation of rhIL-2 Combined with Zhenqi Fuzheng and BaofuKang Suppository in the Treatment of Cervical Intraepithelial Neoplasia II with HPV Infection

Objective: To investigate the clinical effects of recombinant human interleukin-2 (rhIL-2) combined with Zhenqi Fuzheng and Baofukang on cervical intraepithelial neoplasia II (CINII) combined with human papilloma virus infection. Methods: There were 593 patients diagnosed with CINII with HPV infection, including 296 in the control group and 297 in the experimental group. The control group was given only Zhenqi Fuzheng oral and Baofukang suppository vaginal medicine. The experimental group was treated with rhIL-2 injection in addition to Zhenqi Fuzheng oral and Baofukang suppository vaginal medicine which is treated for 3 months. After 3 months, Thinprep cytologic test (TCT), human papilloma virus (HPV) quantitative examination and colposcopy biopsy were reviewed. Results: After 3 months of treatment, the negative conversion rate and total effective rate of HPV in the control group were 58.11% and 70.95% respectively, and the negative conversion rate and total effective rate of the experimental group were 79.46% and 90.57% respectively. There was significant difference between the two groups (p = 0.000). The curative rate of cervical lesions was significantly higher in the test group than in the control group, 89.56%, 68.91%, respectively. The statistical difference between the two groups is significant (p = 0.000). Conclusion: It has an essential clinical value that HPV infection patients and cervical intraepithelial neoplasia II associated with HPV infection patients are treated by rhIL-2 combined with Zhenqifuzheng and Baofukang, which is safe, effective, non-invasive, reusable advantages. However, the long-term efficacy and side effects need to be further studied.

The Prevalence of High Grade Cervical Intraepithelial Neoplasia (CIN) in a Primary Human Papillomavirus (HPV) Cervical Screening Programme Population with HPV Positive and Cytology Negative Smear Results

High-risk HPV is found in 99.7% of cervical cancers. The causative role of HPV in cervical cancer has led to the inclusion of HPV testing as part of cervical screening. A pilot of HPV testing as primary screening was commenced in 2013 at six pilot sites in England. North Cumbria Integrated Care (NCIC) NHS Foundation Trust took part in the pilot, in which women with an HPV-positive/cytology-negative result were recalled at 12 months. Women with HPV type 16/18 found at initial screening and persisting at 12 months in spite of negative cytology were referred to Colposcopy services at 12 months. Women with smear positive for hrHPV other than 16/18 types were recalled twice at 12 and 24 months before referral to colposcopy. Persistent hrHPV positive/cytology negative smear at 12 and 24 months initiated a colposcopy referral. Objective: To assess the prevalence of high grade CIN and invasive cancer in patients referred to colposcopy services at NCIC NHS Foundation Trust with hrHPV positive/cytology negative smears. Method: The study was conducted at NCIC NHS Foundation Trust between January 2015 and December 2017. Data was collected retrospectively from the colposcopy data base (INFOFLEX). All patients with HPV positive/cytology negative smears seen in colposcopy clinic during the study period were included. Patients with high grade CIN, cervical glandular intraepithelial neoplasia (CGIN) or invasive cancer were recorded. Results: 763 women were included in the study. A total of 50 (6.6%) women had high grade CIN, CGIN or invasive cancer. 40 of these 50 women (80%) were treated by large loop excision of the transformation zone (LLETZ). Conclusion: HPV primary screening is more effective than cytology-based screening. A high grade HPV positive result with negative cytology, persisting for one year in type 16/18 and for two years in other high-risk HPV types, warrants referral for colposcopy, as 6.6% of women in this study had high grade or invasive pathology.

Pathological characteristics and immunophenotype analysis of cervical intraepithelial neoplasia

Objective To explore the clinical pathological features and immunophenotypes of cervical intraepithelial neoplasia(CIN).Methods The protein expression of p16, p53, Bcl-2, and c-erb B-2 in 59 cases of CIN, 20 cases of cervical squamous cell carcinoma, and 20 cases of normal cervical tissues were tested using immunohistochemistry staining.Results The expression rates of p16, p53, Bcl-2, and c-erb B-2 in CIN tissues were 76.3%(45/59), 28.85(17/59), 61.0%(36/59), and 40.0%(23/59), respectively. The expression rates of p16, p53, Bcl-2, and c-erb B-2 in cervical squamous cell carcinoma tissues were 60.0%(12/20), 60.0%(12/20), 75.0%(15/20), and 65.0%(13/20), respectively. The expression rates of p16, p53, Bcl-2, and c-erb B-2 in normal cervical tissues were 0.0%(0/20), 0.0%(0/20), 35.0%(7/20), 0.0%(0/20), respectively. In comparison to normal tissues, the differential expressions of p16, p53, and Bcl-2 in squamous cell carcinoma and CIN were statistically significant(P <0.001), whereas the difference between the squamous cell carcinoma and CIN was not significant(P >0.05). In comparison to normal tissues, the differential expressions of c-erb B-2 and p53 in squamous cell carcinoma and CIN were statistically significant(P >0.05). The differential expressions of Bcl-2, c-erb B-2, and p53 in CIN 3 were statistically significant in comparison to CIN 1 and CIN 2(P <0.05). Conclusion Overexpression of Bcl-2 occurs early in the development of cervical cancer, whereas p16 and c-erb B-2 overexpression are markers for cell malignancy. The expression of p53 is correlated with the development of cervical cancer.

The Role of IL-2, IL-4, IL-10 and IFN-γ Cytokines Expression in the Microenvironment of Cervical Intraepithelial Neoplasia

Objective: To investigate the role of IL-2, IL-4, IL-10 and IFN-γ cytokines in the microenvironment of cervical intraepithelial neoplasia. Methods: 180 patients participating were enrolled in this trial, where 50 cases are in control group, 50 cases are in low-grade squamous intraepithelial lesion (LSIL) group, 50 cases are in high-grade squamous intraepithelial lesion (HSIL) group, and 30 cases have cervical carcinoma. ELISA methods are used to detect the expression of IL-2, IL-4, IL-10 and IFN-γ in the lavage fluid, and all data is analyzed using one-way analysis of variance. Results: The expression of IL-2 and IFN-γ in cervical lavage fluid gradually decreases with the aggravation of the lesion. The expression of IL-4 and IL-10 increases by the aggravation of pathological changes. There were statistically significant differences in IL-2 level among the four groups. IFN-γ levels are significantly different between the cervical cancer group and the other groups, also between HSIL and LSIL group, and between HSIL and the control group, but no statistically significant difference was observed in IFN-γ e between LSIL and the control group. The expressions of IL-4 and IL-10 between the cervical cancer group and the other groups were significantly different, and also between HSIL and the control group. But there was no statistically significant difference between LSIL and the control group, HSIL and LSIL. Conclusion: The cytokines of IL-2, IL-4, IL-10 and IFN-γ in the microenvironment of the cervix play an important role in the occurrence and development of cervical intraepithelial neoplasia. This study may provide the evidence for diagnosis and treatment of cervical intraepithelial neoplasia.

Efficacy and Tolerability of Thermocoagulation Treatment of High-Grade Cervical Intraepithelial Neoplasia

Purpose: Cervical cancer is still one of the main causes of cancer and mortality in women, especially in low- and middle-income countries, although it is a completely preventable disease through the detection and treatment of pre-cancer lesions. We evaluated the efficacy and tolerability of thermocoagulation treatment of high-grade cervical intraepithelial neoplasia (CIN 2 and 3). Methods: We evaluated 115 women with high-grade cervical intraepithelial neoplasia, 54 with CIN 2 and 61 with CIN 3, confirmed by biopsy and without previous treatment, from January 2016 to December 2018, undergoing thermocoagulation treatment at the Lower Genital Tract Pathology and Colposcopy Service of the Hospital de Clinicas de Porto Alegre, Porto Alegre, Brazil. Results: The mean age was 33.11 years (SD = 9.83) for CIN 2 and 35.28 years (SD = 7.97) for CIN 3 patients. Treatment efficacy was 90.8% in CIN 2 and 94.9% in CIN 3 cases. Pain was the main symptom reported at the time of treatment, occurring more frequently in the CIN 3 group (49.1% versus 27.8% in the CIN 2 group). As the more important long-term complication, there were 3 cases of pelvic inflammatory disease in CIN 2 (5.6%) and 3 in CIN 3 group (5.0%). The percentage of residual lesion was very low in both groups, 5 in CIN 2 group (9.2%) and 2 in CIN 3 group (3.4%). Conclusion: Thermocoagulation is an effective method for the treatment of cervical pre-cancer lesions (CIN 2 and CIN 3), with a low risk of adverse events and complications.

Photodynamic Therapy for Low-grade Cervical Intraepithelial Neoplasia(CIN1):A Case Report

As a drug-mechanical combination technology,photodynamic(PDT)can achieve accurate and taurgeted therapy for maligmant tumors and benign diseases through the production of reactive oxygen species,oxygen free radicals or singlet oxygen by photosensitizers at specific wavelengths.Compared with traditional surgery,it has the advantages of selective killing.repeatable teatment,preserving target organ fiunction and so on.The purpose of this study was to explore the clinical value of photodynamic therapy in cervical precancerous lesions by taking the patients with low-grade cervical intaepithelial neoplasia(CIN1)with high-risk human papillomavinus(HR HPV)persistent infection diagnosed by"three-step diagnosis and teatment procedure"as an example.Using HiPorfn as a photosensitizer,photodynamic therapy was performed 48 hours after intavenous drip.Set laser wavelength 630nm,light dose density 137.58J/cm^2,ansmission efficiency 1.42,output power 2w.3cm columnar optical fber was placed around the 2cm in the cervical canal to cover all the lesions,and the inadiation time was 900s(600s in the cervical canal and 300s outside the cervical canal).The patients were given oxygen inhalation for 6 hours after operation,and the patients were observed for itching and other discomfort,and paid attention to avoid light.Photodynamic therapy was performed again in the same way on the second day.After two months of treatment,pathological biopsy showed chronic cervicitis,indicating that the disease had been effectively controlled.Theoretically,although the patient is not the absolhute indication of photodynamic therapy(hat is,meeting CIN Ⅱ or CIN Ⅲ,having fertility requirements and not undergoing surgery),this therapy can remove not only the superficial lesions inside and outside the cervix,but also the potential lesions not found under colposcopy.It can also block the persistent infection of HPV by.inhbting the expression of HPV18,E6 and E7mRNA in Hela cells.In combination with Baofukang suppository,it can block HPV infection.Increase the negative conversion rate of cervical HPV and reduce the probablity of recurence after CIN1 cure.For youmng female patients with persistent HR-HPV infection and fertility requirements,photodynamic therapy is an effective choice for clinical treatment of CIN1.

Pregnancy and Obstetrical Outcomes Following Treatment for Cervical Intra-Epithelial Neoplasia (CIN) in Two Hospitals of a Low-Resource Country

Objective: In Cameroon, more than 80% of women suffering from cervical intraepithelial neoplasia (CIN) are within the reproductive age. This study intended to analyze the pregnancy outcomes following cervical treatment in Cameroon. Methods: This was a cohort study with two years retrospective data collection involving 82 women who underwent cervical treatment for CIN in two Cameroonian hospitals from January 2015 to December 2017. Data were collected from CIN treatment to end of pregnancy where applicable. Data analysis was done using Epi Info software version 3.5.4. Results: We collected data from 82 patients aged 27 to 48 years, with a mean age of 36.5 (SD: 5.3) years. During the study period, 33 out of 82 participants became pregnant 40.2 [29.56 - 51.66]%. The factor associated with pregnancy occurrence after treatment was age less than 35 years (Odds ratio = 4.37 [1.7 - 11.2]. From the 33 pregnancies recorded, 17 (51.5%) ended in a delivery, amongst which 15 (88.2%) were vaginal. Conclusion: Pregnancy frequency over two years following cervical treatment for CIN was relatively good, and younger women (age 35 years) were significantly more like to have conceived compared to their older counterparts. Post-treatment delivery outcomes seem to be similar to those in the general population.

Gain of human telomerase RNA gene is associated with progression of cervical intraepithelial neoplasia grade Ⅰ or Ⅱ

Background The 3q26 chromosome region, where the human telomerase RNA gene （hTERC） is located, is a biomarker for cervical cancer and precancerous lesions. The aim of this study was to confirm the value of measuring hTERC gene gain in predicting the progression of cervical intraepithelial neoplasia grade Ⅰ or Ⅱ （CIN-Ⅰ and -Ⅱ, respectively） to CIN-Ⅲ and cervical cancer. Methods Liquid-based cytological samples from 54 patients with CIN-Ⅰ or CIN-Ⅱ lesions were enrolled in this study. Follow-up was performed with colposcopy and biopsy within 24 months after the diagnosis of CIN-Ⅰ or CIN-Ⅱ. Copy numbers of the hTERC gene were measured by fluorescence in situ hybridization with a dual-color probe mix containing the hTERC gene probe （labeled red） and the control, the chromosome 3 centromere-specific probe （labeled green）.Results All patients whose lesions progressed from CIN-Ⅰ or CIN-Ⅱ to CIN-Ⅲ displayed a gain of the hTERC gene, whereas patients where the hTERC gene was not amplified did not subsequently progress to CIN-Ⅲ or cervical cancer. The signal ratio pattern per cell was recorded as N：N （green： red）. The numbers of cells with the signal ratio pattern of 4：4 or N：≥5 in patients whose lesions progressed to CIN-Ⅲ were significantly higher than those whose lesions did not progress. Significantly, none of the patients with a 4：4 signal ratio pattern regressed spontaneously.Conclusions In conclusion, measurement of hTERC gene gain in CIN-Ⅰ or CIN-Ⅱ patients using liquid-based cytological samples could be a useful biomarker to predict the progression of such cervical lesions. In addition, a 4：4 or N：≥5 signal ratio pattern may indicate the unlikeness of spontaneous regression of CIN-Ⅰ or CIN-Ⅱ lesions.

Value of frozen section examination in diagnosis and treatment of high-grade cervical intraepithelial neoplasia

Background Invasive cancer of the cervix is considered a preventable disease because it has a long pre-invasive state, cervical cytology screening programs are currently available, and treatment of pre-invasive lesions is effective. We tested the accuracy of frozen section examination （FSE） of cone specimens to identify the endocervical margin and rule out invasion in patients with high-grade cervical intraepithelial neoplasia （CIN）. Methods For 320 consecutive patients with a preoperative biopsy result of CIN stage 2/3, cold-knife conization （CKC） was performed followed by FSE. The results from analyses of permanent paraffin sections （PS） were compared with the FSE findings. Results The accuracy of FSE was 87% （278/320）. For all of the seven patients with an invasive squamous cell carcinoma of the cervix identified by FSE, the diagnosis was confirmed by PS analysis. For one patient, the FSE result was cervicitis, whereas PS ananlysis showed microinvasive carcinoma. Appropriate surgery was performed for all patients based on the FSE and biopsy results. The FSE and PS results were not significantly different （P=-0.000）. Definitive examination of margin status using PS was concordant with FSE findings in all cases. Conclusions FSE is a rapid and reliable method for evaluating CKC specimens. It can identify frank invasion, permit adequate treatment in a one-stage procedure, and reliably detect clear resection margins. Since discrepancies do exist and may result in inappropriate treatment, further research is required to decrease these discrepancies and avoid missing even one case.

Predictors and clinical significance of the positive cone margin in cervical intraepithelial neoplasia III patients

Background Conization is being widely accepted for diagnosis and treatment of cervical intraepithelial neoplasia （CIN）. There is controversy as to which factors are most predictive of a positive cone margin and the clinical significance of it. We conducted this study to identify the predictive factors and to evaluate the clinical significance of a positive cone margin in CIN III patients.Methods A retrospective review was conducted of 207 patients who had undergone conization due to CIN III from January 2003 to December 2005 at Peking Union Medical College Hospital. Of these, 67 had a subsequent hysterectomy Univariate and multivariate analysis were utilized to define the predictive factors for a positive cone margin, and to compare the pathologic results of conization with subsequent hysterectomy.Results One hundred and fifty-one （72.9%） were margin free of CIN I or worse, 37 （17.9%） had CIN lesions close to the margin and 19 （9.2%） had margin involvement. A total of 56 cases （27.1%） had positive cone margins （defined as the presence of CIN at or close to the edge of a cone specimen）. Univariate analysis showed that the parity, cytological grade multi-quadrants of CIN III by punch biopsy, gland involvement, as well as the depth of conization were significant factors correlated with a positive cone margin （P 〈0.05）. However the age, gravidity, grade of dysplasia in punch biopsy, as well as the cone methods were not significantly correlated （P 〉0.05）. Multivariate analysis revealed that the cytological grade （OR=1.92）, depth of conization （OR=2.03）, parity （OR=3.02） and multi-quadrants of CIN III （OR=4.60） were significant predictors with increased risk for positive margin. The frequency of residual CIN I or worse in hysterectomy specimens was found to be 55.6% （20/36） in patients who were margin free, 71.4% （15/21） in patients with CIN occurring close to margin, and 80.0% （8/10） in patients with margin involvement. The frequency of residual CIN III or worse was found to be 13.9% （5/36）, 23.8% （5/21） and 50.0% （5/10） respectively in different groups.Conclusions Cytological grade, depth of conization, parity and multi-quadrants of CIN III in punch biopsy were significant factors with increased risk in predicting a positive cone margin. Margin status of conization did not mean the presence or absence of CIN, but rather the varied frequency of residual CIN in specimens of subsequent hysterectomy. In view of this fact, it is suggested that the margin status of conization be a valuable surrogate marker for clinical management of CIN Ⅲ.

Proliferative activity in cervical intraepithelial neoplasia and cervical carcinoma

Objective To detect the proliferative activity of cervical cells in cervical intraepithelial neoplasia (CIN) and cervical carcinoma by using proliferating cell nuclear antigen (PCNA) immunohistochemistry and mitotic index (MI).Methods Sixty specimens of CIN and cervical carcinoma from patients treated at the Obstetrics and Gynaecology Hospital, Shanghai Medical University, were analyzed. Among them, 8 were CIN Ⅰ Ⅱ, 16 CIN Ⅲ, 18 squamous carcinoma, and 18 adenocarcinoma. Serial 4μm sections were prepared for haematoxylin and eosin (HE) staining to observe mitotic figure and pathological grading and for immunohistochemical staining of PACN which was performed by murine monoclonal antibody PC 10 (DAKO PCNA).PCNA index (PI) and MI of CIN Ⅲ and cervical carcinoma were significantly higher than those of normal cervix and CIN Ⅰ Ⅱ. PI and MI increased with progress of disease. In addition, higher level of PI was correlated with higher level of MI, and higher levels of PI and MI were related with higher histological grade. However, there was no significant relationship between PI, MI, clinical staging, and lymph node metastases.PCNA and mitotic figure are useful for evaluating proliferative activity in CIN and cervical carcinoma, and could be used to distinguish CIN Ⅲ from CIN Ⅰ Ⅱ and evaluate the malignant potential of tumour tissues.

Treatment of Cervical Intraepithelial Neoplasia with Microwave Coagulation.

From Nov. 1990 to Dec. 1993, 128 patients with histologically diagnosed cervical intraepithelial neoplasia (CIN) grade 1 or 2, were randomized treated with the follow-

Expression and Significance of SHP-2 in Human Papillomavirus Infected Cervical Cancer

This study investigated the expression and prognostic value of SHP-2 in cervical cancer caused by human papillomavirus (HPV) infection. Forty-five specimens from patients with cervical cancer (stageⅠ-Ⅲ), 32 specimens from patients with cervical intraepithelial neoplasia (CIN) (Ⅰ, Ⅱ) and 20 normal cervical samples from patients with hysteromyoma were collected in Department of Pathology for comparison. The expression levels of SHP-2 and IFN-β proteins were detected by using immunohistochemistry. The mRNA expression level of SHP-2 was detected by using quantitative real-time polymerase chain reaction (PCR). HPVs were detected by HPV GenoArray Test. The Spearman correlation was used to compare the expression level of SHP-2 in HPV infected cervical cancer vs non-HPV infected normal cervix. The level of SHP-2 protein expression in the cancer tissues (88.8%) was significantly higher than in CIN tissues (62.5%) and normal cervixes (45%) (P<0.05 and P<0.05, respectively). The SHP-2 mRNA levels in the cancer tissues were upregulated as compared with those in the normal cervixes (P<0.05). Twenty-one (46.7%) cervical cancers, 25 (78.1%) CINs and 17 (85%) normal cervixes showed IFN-β positive staining in cytoplasm. There was statistically significant difference in the expression rate of IFN-β between cervical cancer and normal cervix (χ2=8.378, P<0.05) as well as between cervical cancer and CIN (χ2=7.695, P<0.05). HPV16/18 infections could be found in normal cervixs (15%), CINs (68.7%) and cervical cancers (84.4%). There was a correlation between HPV infection and SHP-2 expression in cervical cancer (rs=0.653, P<0.05). SHP-2 may be a useful prognostic and diagnostic indicator for HPV infected cervical cancer. In cervical cancers, SHP-2 mRNA and protein overexpression was associated with IFN-β lower-expression.

Photodynamic therapy with TBZPy regulates the PI3K/AKT and endoplasmic reticulum stress-related PERK/eIF2α pathways in HeLa cells

Background:((1-triphenylaminebenzo[c][1,2,5]thiadiazole-4-yl)styryl)-1-methylpyridin methylpyridin-1-ium iodide salt(TBZPy)is a novel photosensitizer that displays excellent photodynamic properties.However,There are few reports on the mechanism of action of the TBZPy photodynamic.Previous studies revealed that photodynamic therapy(PDT)could induce endoplasmic reticulum stress by acting on the endoplasmic reticulum.Therefore,in this study,we investigated the effects of endoplasmic reticulum stress induced by TBZPy-PDT in treating High-risk human papillomavirus(HR-HPV)infection and their underlying mechanisms.Methods:The human cervical cancer cell line HeLa(containing whole genome of HR-HPV18)was treated with TBZPy-PDT.Cell migration,invasion,and colony-forming ability were evaluated using wound-healing,Transwell invasion,and colonyforming assays,respectively.Through western blot analysis,we determined the level of expression of the PI3K/AKT and PERK/eIF2αpathway proteins and the proteins associated with calcium trafficking and apoptosis.The calcium levels in the cytoplasm were detected via flow cytometry.Results:The result shows that TBZPy-PDT could inhibite the migration,invasion,and colony forming ability of infected HeLa cells by downregulating the PI3K/AKT pathway in vitro.And we found that TBZPy-PDT induced endoplasmic reticulum stress-specific apoptosis via the PERK/eIF2αpathway.Moreover,TBZPy-PDT increased the levels of calcium and calmodulin,while decreasing the levels of endoplasmic reticulum calcium-binding proteins.Conclusions:TBZPy-PDT is effective on treating human papillomavirus-infected cells.Targeting the PI3K/AKT and PERK/eIF2αpathways and the endoplasmic reticulum stress process may help improve the effects of TBZPy-PDT for treating high-risk human papillomavirus infection.

Risk factors for the development of vaginal intraepithelial neoplasia

Background Vaginal intraepithelial neoplasia （VAIN）, as a precursor of vaginal cancer, is a rare disease. Its prevalence has not been well analyzed. This research is to ascertain the risk factors for VAIN in a Chinese population. Methods A case-control study was conducted, including 63 VAIN cases and 64healthy controls. In all subjects Pap smear and HPV tests were performed. A questionnaire survey was distributed, covering information on socio-demographic characteristics, smoking, past history, reproductive and sexual histories. The clinical pathological data were collected from medical records including symptoms, Pap smear results, grade of lesions, and human papillomavirus （HPV） status. Results Postmenopausal women had a 2.09 times higher risk for VAIN than pre-menopausal women （95% CI： 1.10-3.85; P=-0.024）. The patients with previous hysterectomy had an increased risk of VAIN （OR=4.69; P=0.003）. Patients with a history of cervical cancer or CIN were predisposed to VAIN （OR=78.75; P 〈0.0001）. The rate of HPV infection in VAIN was significantly higher than in controls, and an increased risk of VAIN was observed in patients with higher viral load （OR=126.00; P=0.000）. Multivariate analysis showed that HPV infection and a history of CIN or cervical cancer were still found to be significant in patients. Conclusion HPV infection and a history of CIN or cervical cancer are the main risk factors for the development of VAIN.

Value of P16 expression in the triage of liquid-based cervical cytology with atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesions

Background The management of atypical squamous cells of undetermined significance/low-grade squamous intraepithelial lesions （ASCUS/LSIL） is still controversial and it is advisable to make a triage for these two cytological abnormalities. P16INK4 （P16） has been shown to be a potential biomarker for predicting high-grade cervical intraepithelial neoplasia （CIN） and cervical cancer. The aim of the study was to determine the value of P16 expression by immunostaining method compared with high-risk human papillomavirus （HR-HPV） DNA test in the triage of ASCUS/LSIL women. Methods Totally 86 eligible residual liquid-based cytological specimens with ASCUS and 45 with LSIL were obtained. All specimens were submitted to HR-HPV DNA test （HC2） and P16 immunocytochemical staining simultaneously. And all women underwent colposcopy and biopsy after cytology. Results The positive rate of P16 staining was 32.6% in ASCUS and 42.2% in LSIL, which was significantly lower than that of HR-HPV test in both ASCUS （P〈0.05） and LSIL （P〈0.05）. Moreover, the positive rate of P16 staining was 12.7% in normal histology, 61.5% in CIN 1,87.0% in CIN 2-3, and 100.0% in cancer, in which P16 positive rate was significantly lower than HR-HPV positive rate in normal group. The sensitivity, specificity, positive predictive value （PPV）, negative predictive value （NPV） and accuracy of P16 staining for predicting CIN 2 or more were 87.5%, 68.6%, 38.9%, 96.0%, and 72.1%, respectively in the ASCUS; while 90.0%, 71.4%, 47.4%, 96.2% and 54.7%, respectively in the LSIL, in which the specificity and accuracy of P16 staining were significantly higher than those of HR-HPV test in both ASCUS and LSIL （P 〈0.05）. Conclusion P16 immunostaining had significantly higher specificity and accuracy than HR-HPV DNA test for predicting for high-grade CIN and cervical cancer in ASCUS and LSIL and can be used for the triage of women with ASCUS/LSIL cytological abnormality.

Clinical Outcome of Topical Interferon Alpha-2b Cream in Phase II Trial for LSIL/CIN 1 Patients

Objectives: Interferon alpha-2b possesses variable activity against human papillomavirus (HPV) associated cervical intraepithelial neoplasia (CIN). No topical therapy is currently available for treatment of early stage CIN. We evaluated a new patented drug delivery technology in order to achieve topical efficacy. Methods: Two separate studies were conducted in parallel. IFN002 (treatment group) was an open label study. Twenty patients with Pap IIW, III and IIID (CIN1) were treated with intravaginal application of Interferon alpha-2b cream (5 g, 2 MIU/g) three times a week (alternate days) for 6 weeks with 6 weeks of follow up to determine its effect on cytologic and colposcopic assessment. HPV001 (control group) was a 12 week observational study. Both studies had similar inclusion/exclusion criteria and patient population. Results: In IFN002, 8 of 20 patients (40%) in the ITT population showed resolution of abnormal Pap smear during the 12 weeks following start of treatment (responders). In HPV001, 7 of 21 patients (33.3%) were regressors (p = 0.45, one-sided FET). In the PP population, 7 of 12 (58.3%) patients in IFN002 were regressors com-pared to 7 regressors of 19 patients (36.8%) in HPV001 patients (p = 0.21, one-sided FET). Among patients with Pap IIID, 8 of 14 patients in IFN002 showed resolution of abnormal Pap smear, while 4 of 14 patients resolved in HPV001 (one-sided FET, p = 0.13). Conclusions: Interferon alpha-2b cream (5 g, 2 MIU/g) may be an effective treatment for CIN 1 patients, and future investigation is warranted.