[Evaluation of variable methods for HPV testing].

OBJECTIVE: To evaluate clinical efficacy of different HPV methods in screening of cervical cancers. METHODS: Between August 2011 and November 2011, 424 women in the China-Japan Friendship Hospital were enrolled in this study. All participants were undergone liquid-based cytology test (LCT), Hybrid capture II (HC-II) and real-time (RT) PCR high risk HPV DNA test for HPV16 and HPV18 genotyping. Those results were classified into two group: 424 women at HC-II group with LCT and HC-II test and 421 women at PCR group with LCT and PCR test. All women with atypical squamous cell of undetermined significance (ASCUS) or above in cytological result with high risk HPV positive at two group underwent cervical biopsy by colposcopy.In the mean time, women with negative in cytological results and with HPV 16 and(or) HPV 18 positive also underwent histo-pathological examination by and colposcopy. The results in two groups were discussed:LCT+HC-II group (424 patients) and LCT+PCR12+2 group (421 patients). RESULTS: (1) There was no significant difference in cervical intraepithelial neoplasia (CIN) II or above disease between LCT+HC-II group and LCT+PCR12+2 group (χ(2) = 3.35, P > 0.05).Sensitivity, specificity, positive predictive value and negative predictive value for detection of CINII or above using HC-II and PCR12+2 were 77.8%, 79.4%, 20.4%, 98.1% and 96.3%, 78.2%, 23.2%, 99.7%, respectively. (2) In LCT+PCR12+2 group, it was found 34 women with HPV16 positive, 5 women with HPV 18 positive including 1 women combined with HPV 16 positive, 74 women with other high risk HPV positive and 309 women with HPV negative. Compared to the infection of other high-risk HPV types, HPV 16 and HPV 18 infection leads to a higher chance of cervical lesions with CIN II or above [51.3% (20/39) and 8.1% (6/74) ]. (3) A significant difference of causing cervical cancer and CINII or above was found among women who were infected with HPV 16 and/or HPV 18 infection, with other high-risk HPV types and negative in high-risk HPV infection (χ(2) = 93.98, P < 0.01). CONCLUSION: LCT combined with PCR genotyping HPV could identify CIN II or above disease efficiently.

Sensitive HPV genotyping based on the flow-through hybridization and gene chip.

Persistent infection of high-risk human papillomavirus (HPV) has been recognized as the direct cause of cervical carcinoma. Therefore, detection and genotyping of HPV are important to cervical-cancer screening. In this study, we have evaluated the efficacy of flow-through hybridization and gene chip (HybriMax) on HPV genotyping through comparison of the results with Hybrid Capture II (HC-II) and in situ hybridization (ISH). 591 women were classified into 6 groups according to their histological diagnoses. The overall accordance rate on 13 types of HPV genotypes between HybriMax and HC-II were 92.5% and 100% in the cancer group. The overall accordance was excellent with the Kappa index (KI) of 0.814. The value of KI in each group was 0.750 (normal cytological diagnosis), 0.781 (chronic cervicitis), 0.80 (condyloma acuminatum), 0.755 (cervical intraepithelial neoplasia (CIN) I), 0.723 (CIN II), and 0.547 (CIN III) (0.75 > KI > 0.4, good; KI ≥ 0.75, excellent). The 10 most common HPV subtype detected by HybriMax were 16, 52/58, 18, 33, 31, 81, 53, 68, and 66 in patients, and 16, 68, 18, 52, 58, 11, 53, 31/39, and 33 in normal controls. In conclusion, HybriMax is an efficient method for HPV genotyping and more suitable for clinical use.

Value-based medicine analysis on loop electrosurgical excision procedure and CO2 laser vaporization for the treatment of cervical intraepithelial neoplasia 2.

AIM: The best treatment option for cervical intraepithelial neoplasia 2 (CIN2) is controversial and there is a lack of studies in value-based medicine. This multicenter comparative study was undertaken to evaluate the effectiveness, cost-effectives and quality of life (QOL) of loop electrosurgical excision procedure (LEEP) and CO(2) laser vaporization for the treatment of CIN2. MATERIAL AND METHODS: A database of LEEP and laser vaporizations performed at three research centers was created. Patients with colposcopic-histopathologically confirmed CIN2 were randomly submitted to LEEP and laser vaporization. Cytology, human papilloma virus (HPV) DNA test and histology were performed, and a questionnaire on QOL was filled out during follow-up. Effectiveness, cost-effectives and QOL were analyzed. RESULTS: Three hundred and thirty-eight women with CIN2 were included in the study. Frequencies of remission, and persistent and recurrent CIN were 89.2%, 7.2%, and 3.6% for LEEP, and 86.7%, 12.6%, 0.70% for laser, respectively. There was no significant difference in remission and persistence of CIN. There was a significant difference in the number of operations, recovery time and costs. Women treated with two methods showed relatively identical QOL. CONCLUSION: Both LEEP and CO(2) laser vaporization are effective and reliable treatments for CIN2, whereas cervical tissue can be obtained for histology by LEEP. Preoperative evaluation and postoperative follow-up are important. Gynecologists should pay attention to QOL of patients with CIN.

[Pregnancy related cervical cytological changes and clinical management].

OBJECTIVE: To investigate characteristics of cervical cytology and management in pregnant women. METHODS: From Aug. 2006 to Jan. 2010, 5152 pregnant women who received antenatal and postpartum examination underwent cervical cytological screening by liquid-based cytological test (LCT) in China-Japan Friendship Hospital. The cytological diagnosis was in accordance with the Bethesda system (TBS) 2001 diagnosis and classification system. The abnormal LCT results were followed up at 3 months after postpartum. The diagnosis of high-grade squamous intraepithelial lesions (HSIL) and squamous cell carcinoma (SCC) were based on colposcopic examination and biopsy during pregnant. The diagnosis of atypical glandular cells (AGC) was based on curettage and biopsy at postpartum 6 weeks. The histopathology of biopsy were compared and analyzed. RESULTS: (1) Cervical cytological changes related with pregnancy: among 5152 cases, it was found navicular cells in 3215 cases (62.40%), decidual cells in 783 cases (15.20%), reactive glandular cells in 369 cases (7.16%), and trophoblastic cells in 55 cases (1.07%). (2) LCT results: among 5152 cases, the normal samples were 4125 cases (80.07%), the inflammatory samples were 542 cases (10.52%), and the samples of abnormal epithelial cells were 485 cases (9.41%). Among those abnormal cases, 291 cases (5.65%) were in atypical squamous cells (ASC), 153 cases (2.97%) were in low-grade squamous intraepithelial lesions (LSIL), 33 cases (0.64%) were in HSIL, 1 case (0.02%) were in SCC and 7 cases (0.14%) were in AGC. (3) Histological pathology results: all women with HSIL and SCC underwent colposcopic examination and biopsy, it was found 28 cases in cervical intraepithelial neoplasia (CIN)II-III, 1 cases in adenosquamous carcinoma. 7 women underwent curettage and biopsy at postpartum 6 weeks which were diagnosed by AGC, the histopathological diagnosis was all negative. The concordance rate of cytopathologic and histopathologic diagnosis was 71% (29/41). (4) FOLLOW-UP: 485 women with abnormal LCT results were all followed up to 3 months at postpartum. Women with HSIL, SCC and AGC undergoing biopsy showed normal LCT results during follow-up. Those women with ASC and LSIL did not undergo colposcopic examination and biopsy. The regression rate was 72.3% (321/444) at postpartum 3 months. CONCLUSIONS: The navicular cells were primarily morphological characteristics of cytology during pregnant and postpartum women. Some changes were easily confused with malignant lesions. It should be careful discrimination, and avoid excessively diagnosis and misdiagnosis. It suggested that we should follow up those women closely and expand the indication of colposcopic biopsy.

[Comparative study on the effects of LEEP and laser CO(2) vaporization in cervical intraepithelial neoplasia II].

OBJECTIVE: to compare the effect and complications of loop electro-surgical excision procedure (LEEP) and laser CO(2) vaporization in the treatment of cervical intraepithelial neoplasia II. METHODS: a total of 338 CINII women were recruited into this multi-center comparative study. The diagnosis was confirmed by histopathological examination for cervical epithelial cell abnormalities. And colposcopic examination was submitted to LEEP (n = 195) or laser CO(2) vaporization (n = 143) respectively. A post-treatment follow-up of 3, 6 and 12 months was carried out to compare the effect of two methods. RESULTS: among 195 women undergoing LEEP, the frequency of cure, persistent and recurrent CIN was 89.2% (n = 174), 4.1% (n = 8) and 3.6% (n = 7) respectively. And among 143 women receiving laser CO(2) vaporization, the frequency of cure, persistent and recurrent CIN was 86.7% (n = 124), 4.9% (n = 7) and 0.70% (n = 1) respectively. There was no statistical difference in cure rates, persistence or recurrence of CIN (P > 0.05). The recovery time, the operative frequency and intra-operative blood loss were significantly different in two groups. CONCLUSION: both LEEP and CO(2) vaporization are both effective and reliable for the treatment of cervical intraepithelial neoplasia II. However, pathological specimens may be harvested during LEEP. It is of vital importance to conduct preoperative colposcopic assessment and standard postoperative follow-ups.

Human papillomavirus type-distribution in cervical cancer in China: the importance of HPV 16 and 18.

Prophylactic vaccination against HPV 16 and 18 has the potential for effective prevention of high-grade precancer (cervical intraepithelial neoplasia [CIN)] 2/3) and ICC caused by these viruses (globally 50 and 70%, respectively) when employed in women prior to starting sexual activity. To provide data for decisions on HPV vaccination in China, we determined HPV type-distribution in ICC and CIN 2/3 from women of different regions within China. A multicenter study was conducted by randomized sampling of paraffin blocks of 664 ICC (630 squamous cell carcinoma [SCC]; 34 adenocarcinoma [ADC]), 569 CIN 2/3 cases from seven regions of China. Histological diagnosis was confirmed in 1,233 cases by consensus review. HPV DNA was detected using the SPF10 LiPA25 version 1 assay. HPV prevalence was 97.6% in SCC, 85.3% in adenocarcinoma, and 98.9% in CIN 2/3. HPV 16 (76.7%) and HPV 18 (7.8%) were the most common, together accounting for 84.5% of SCC, followed by HPV 31 (3.2%), HPV 52 (2.2%), and HPV 58 (2.2%). HPV positivity in SCC did not differ notably by region. However, SCC cases from women

A study on the mechanism of 5-aminolevulinic acid photodynamic therapy in vitro and in vivo in cervical cancer.

5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is a new ablation treatment for tumors, while its function mechanism in cervical cancer has not been elucidated. In this study, we investigated the effects of ALA-PDT on the cervical cancer cell line Me180, to search for optimal parameters of PDT. ALA-PDT on proliferation of Me180 was examined by MTT assay to find the optimal function parameters of ALA-PDT. Apoptosis was observed by using AnnexinV-FITC/PI double staining, Hoechst 33342 staining and May-Grünwald-Farbstoff Giemsa staining. Furthermore, we established a tumor model and 6 mice of each group underwent measurement of the tumor size on days 3, 7, 14, and 21 after treatment. The mRNA expression of survivin, bcl-2, p53, bax and bad in Me180 cells were detected by real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) in vitro and in vivo. Finally, we compared the effects between topical and intravenous administration. Based on the above studies, we found ALA-PDT induced apoptosis and G0-G1 phase arrest of Me180 cells. The tumor volume of the topical administration and PDT group was the smallest at 7-14 days post-PDT. H&E staining showed remarkable subcutaneous necrosis in the PDT groups. The mRNA expression of survivin and bcl-2 in Me180 cells were suppressed post-PDT. Topical administration of PDT is recommended in treating cervical cancer so as to minimize the side-effects and inconvenience of phototoxic reaction brought by PDT. Our data may contribute to making the mechanism of PDT on cervical cancer clearer and give some useful suggestions for clinical application.

[Detection of multiple human papillomavirus infection in cervical specimens by flow fluorescent hybridization].

OBJECTIVE: To explore the clinical significance of detection of multiple human papillomavirus infection in cervical lesions detected by flow fluorescent hybridization technology with Luminex multi-analytic profiling (xMAP). METHODS: Cervical exfoliated cell specimens were collected from 301 randomly selected women accepting opportunistic screening for cervical lesions with the cytological results and hybrid capture 2 (HC-2) assay>or=atypical squamous cells of uncertain significance (ASC-US), 48 with the pathological diagnosis>or=cervical intraepithelial neoplasia (CIN)2 (case group) and 253 with normal histological result or only inflammation (control group), aged (34+/-9) (21-59). The samples were tested with xMAP technology with blind method. The coincidence of the xMAP and HC-2 was assessed. The HPV genotype, multiple HPV infection rate, and their relationships to the patients' clinical-pathological features were analyzed. RESULTS: The rates of sensitivity, specificity, and accuracy of xMAP technology to detect>or=CIN2 cervical lesions were 80.49%, 80.00%, and 80.07% respectively. The positive and negative predictive values were 47.06% and 96.30% respectively. The Kappa Index for agreement between xMAP technology and HC-2 was 0.56. The prevalence rate of high-risk HPV infection was 28.24% (85/301). The prevalence rate of multiple HPV infection was 11.30% (34/301), significantly lower than that of single type high-risk HPV infection (16.94%, P<0.05). The proportion of multiple HPV infection in total positive HPV results was 35.05% (34/97). The proportion of duplex and treble HPV infection were 29.90% (29/97) and 5.15% (5/97) respectively. The multiple HPV infection rate of the case group was 37.50% (18/48), significantly higher than that of the control group (6.32%, 16/253, P<0.01). The common duplex HPV infection modes were 16+51/58 (n=4), 51/58+52/59 (n=4), 11+16 (n=3), and 11+52/59 (n=3), 18+52/59 (n=3). The common treble HPV infection modes were 11+16+52/59, 16+18+31, 16+18+52/59, 31+33+39, and 31+33+52/59 (all n=1). HPV16, 52/59, 51/58, 18, 11, and 31 were the common types in multiple HPV infections. CONCLUSION: Flow fluorescent hybridization technology is able to detect multiple HPV infection that is associated with cervical lesions and to identify the HPV genotypes.

[Detection of human papillomavirus L1 capsid protein expression in liquid-based cytology samples with abnormal cytology.].

OBJECTIVE: To investigate the possibility of detection of the human papillomavirus (HPV) L1 capsid protein to predict the course of mild or moderate cervical intraepithelial neoplasia (CIN). METHODS: Immunocytochemical analysis using antibody against HPV L1 capsid protein was carried out on 274 samples obtained from women performed TriPath Pap tests, positive for high-risk HPV DNA detected by hybrid capture II (HC-II) or cytologic diagnosed atypical squamous cells of undetermined significance (ASCUS) or more severe. For cytological diagnosed, there were ASCUS 105 cases, low-grade squamous intraepithelial lesions (LSIL) 119 cases, atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H) 9 cases, high-grade squamous intraepithelial lesions (HSIL) 36 cases, and squamous cell carcinoma (SCC) 5 cases. But for the pathologic diagnosed, there were chronic cervicitis 96 cases, CINI 85 cases, CINII 55 cases, CIN III 32 cases, and SCC 6 cases. RESULTS: Of the 274 cases, HPV L1 capsid protein was positive in 69.8% (67/96) of cervicitis, 83.5% (71/85) of CINI, 41.8% (23/55) of CINII, 3.1% (1/32) of CINIII and 0(0/6) of SCC. Cytologic diagnosis revealed a higher expression rate in LSIL (75.6%, 90/119) than that in ASCUS (63.8%, 67/105) or in HSIL + SCC (9.8%, 4/41; all P < 0.01). Of 71 cases with ASCUS and LSIL without treated, none of HPV L1 positive cases (0/55) progressed in cytology, while 19% (3/16) of HPV L1 negative cases progressed to ASC-H, HSIL (P < 0.01). CONCLUSION: The expression rates of HPV L1 protein in liquid-based cell specimen is decreased as the cytopathology diagnosis severe degree, which may imply the histopathology diagnosis of cervix, predict the progression of cervical lesion, and help to treat the cases with ASCUS and LSIL.

[Efficacy and safety of drospirenone-ethinylestradiol on contraception in healthy Chinese women: a multicenter randomized controlled trial].

OBJECTIVE: To evaluate the contraception efficacy, mode of bleeding, side effects and other positive effects of drospirenone-ethinylestradiol (Yasmin) in healthy Chinese women. METHODS: This was a multicenter, randomized, control study of 768 healthy Chinese women who consulted about contraception. The subjects were randomized into Yasmin group (30 microg ethinylestradiol plus 3 mg drospirenone, 573 cases) or desogestrel group (30 microg ethinylestradiol plus 150 microg desogestrel, 195 cases) with the ratio of 3:1. Each individual was treated for 13 cycles. Further visits were required at cycle 4, cycle 7, cycle 10 and cycle 13 of treatment Weight, height, body mass index were evaluated at each visit. The menstrual distress questionnaire (MDQ) was given to the women at baseline, visit 3 (cycle 7) and visit 5 (after cycle 13). RESULTS: The values of basal features were similar between two groups (P > 0.05). The Pearl index (method failure) of Yasmin was 0. 208/hundred women year which was lower than that of desogestrel (0. 601/hundred women year). The mode of bleeding was similar between two groups after trial without showing any significant difference. According to MDQ subscale, the improvement of water retention and increasing appetite during inter-menstrual period and water retention and general well-being during menstrual period in the Yasmin group (-0.297, -0.057, 0.033, 0.150 respectively) was more obvious than that in the desogestrel group (-0.108, 0.023, 0.231, -0.023 respectively) with a significant difference (P < 0.05). Some other values which improved in both two groups, especially the improvement of breast tenderness and pain and skin abnormality in Yasmin group (18.0%, 89/494; 12.6%, 62/494) was more distinct than that in desogestrel group (11.3%, 19/168; 5.4%, 9/168). The mean weight increased in desogestrel group (0.57 kg) while it decreased in Yasmin group (-0.28 kg) with a significant difference (P < 0.01). CONCLUSIONS: Both Yasmin and desogestrel have good efficacy on contraception and similar modes of menstrual bleeding. Yasmin is better than desogestrel in terms of weight control and premenstrual syndrome of oral contraceptive.

[Effects of 5-aminolevulinic acid photodynamic therapy on cervical cancer: in vivo experiment with nude mice].

OBJECTIVE: To study the effects of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) on human cervical cancer, and to identify the best administration regimen with the least phototoxic reaction. METHODS: Seventy-two BALB/c nude mice underwent subcutaneous injection of human cervical cancer cells of the line Me180 so as to establish animal models, and then randomly divided into 6 equal groups: pure topical administration group (undergoing injection of ALA 60 mg/kg around the tumor), pure intravenous administration group (undergoing injection of ALA 250 mg/kg into the caudal vein), topical administration and PDT group (undergoing injection of ALA 60 mg/kg around the tumor and radiotherapy of 630 nm He-Ne laser 6 h after the drug administration), intravenous administration and PDT group (undergoing injection of ALA 250 mg/kg into the caudal vein and radiotherapy of 630 nm He-Ne laser 6 h after the drug administration), and control group (undergoing none of the treatment). 24 h later 6 mice from each group were killed with their tumors taken out. On the days 3, 7, 14, and 21 after treatment the remaining mice underwent measurement of the tumor size. HE staining and pathological examination were performed. Immunohistochemical study was conducted to detect the protein expression of the apoptosis-inhibiting genes of survivin and vascular endothelial growth factor (VEGF). RT-PCR was used to detect the mRNA expression of the apoptosis-inhibiting gene of Bcl-2 and apoptosis-promoting genes of P53, Bax, and Bad. RESULTS: The tumor volumes after treatment of the pure topical administration and pure caudal vein administration groups were (0.09 +/- 0. 02) cm3 and (0.14 +/- 0.04) cm3 respectively, both significantly smaller than that of the control group [(0.67 +/- 0.06) cm3, both P < 0.01]. The tumor volume of the pure topical administration group 7-14 days after treatment was the smallest. HE staining showed remarkable subcutaneous necrosis in the 2 PDT groups. Immunohistochemistry showed remarkable down-regulation of protein expression of survivin and VEGF in the PDT groups. RT-PCR showed that the mRNA expression levels of survivin and Bcl-2 were both significantly lower than that of the control group (both P < 0.01), and the mRNA expression levels of P53, Bax, and Bad were higher than those of the control group, however, not significantly (all P > 0.05). CONCLUSION: ALA-PDT is effective in treatment of cervical cancer. Topical ALA administration + PDT is recommended in treating cervical cancer so as to minimize the side-effects and inconvenience of phototoxic reaction brought by PDT.

[Application of high-risk human papillomavirus testing in women with abnormal cytology].

OBJECTIVE: To detect the high-risk human papillomavirus (HPV) infectious condition in women with abnormal cytology and evaluate its values in the screening of high grade squamous intraepithelial lesion. METHODS: We used hybrid capture 2 (hc2) method to examine 949 patients with abnormal cervical cytology results [ > or =atypical squamous cells of undetermined significance (ASC-US) according to the 2001 The Bethesda System diagnosis criteria]. All subjects also received colposcopy for tissue studies. RESULTS: Among 949 patients with abnormal cytology, the diagnoses of atypical squamous cells (ASC), low grade squamous intraepithelial lesion (LSIL), and high grade squamous intraepithelial lesion (HSIL) were made in 432, 310, and 207 patients, respectively. The high-risk HPV positive rate in ASC, LSIL, and HSIL were 40.3%, 44.8%, and 89.4%, respectively. The numbers of patients with pathologically confirmed results of negative intraepithelial lesion or malignancy (NILM), cervical intraepithelial neoplasia 1, 2, 3 (CIN 1, 2, 3), and squamous cell carcinoma (SCC) were 335, 388, 118, 101, and 7, and the high-risk HPV positive rate was 17.3%, 66.2%, 92.4%, 97.0%, and 100%, respectively. Among patients with atypical squamous cells of undetermined significance (ASC-US), rate of HSIL in high-risk HPV positive group and negative group were 10.2% and 0.8%, respectively (P < 0.01). In screening HSIL, the sensitivities of cytology [ > or = ASC cannot exclude HSIL (ASC-H)] and cytology ( > or = ASC-H) plus high-risk HPV testing were 0.925 and 0.991, and the specificities were 0.510 and 0.748, respectively (P < 0.01). Sensitivitives of cytology ( > or = LSIL) and cytology (> or = LSIL) plus high risk HPV in detecting HSIL were 0.898 and 0.982, respectively, while the specificitives were 0. 567 and 0.779, respectively (P < 0.01). CONCLUSIONS: The positive rate of high-risk HPV increases with the gravity of cervical lesions. In patients with abnormal cervical cytology, high-risk HPV testing can improve the sensitivity and specificity in the screening of HSIL.

[Clinical evaluation of the xMAP technology in detection of high-risk human papillomavirus].

OBJECTIVE: To evaluate the clinical application value of flexible multi-analyte profiling (xMAP) technology in detecting high-risk human papillomavirus (HR-HPV). METHODS: Totally 1 061 women, aged 21-65 years, were randomly enrolled into the study. Cervical exfoliated cells were used in xMAP technology and hybrid capture II (hc2). Pathological diagnosis was used as golden standard. Consistency of these two methods was assessed. RESULTS: The sensitivity and specificity of xMAP technology were 80.31% and 85.83%, respectively. The positive and negative predictive values were 44.5% and 96.9%, respectively. The Kappa value for consistency between xMAP technology and hc2 was 0.58. CONCLUSIONS: The specificity of xMAP technology is similar to hc2 test, but the sensitivity is inferior to hc2. However, these two methods show good consistency in the detection of HR-HPV.

Distribution of human papillomavirus genotypes in cervical lesions.

The aims of the present study were to investigate the distribution of human papillomavirus (HPV) genotypes in cervical lesions, and the association between different HPV genotypes and cervical lesions. Between January 2013 and June 2014, the HPV type determinations of nucleic acid by use of fluorescence polymerase chain reaction (PCR) method of 15,192 outpatients in China-Japan Friendship Hospital were performed and the infection status was analyzed. The results showed that: i) 2,366 Cases were HPV positive and 12,826 cases were HPV negative, the overall infection rate was 15.57% (2,366/15,192), in which a single genotype of HPV infection rate was 11.63% (1,767/15,192), and multiple genotypes of HPV infection rate was 3.94% (599/15,192); ii) HPV16, HPV52 and HPV58 infections were the most common HPV genotypes, the infection rates were 3.95% (600/15,192), 2.86% (435/15,192) and 2.67% (406/15,192), respectively; and iii) According to the gold standard of histopathological analysis via hematoxylin-eosin staining, HPV16, HPV52 and HPV58 accounted for 58.80% (154/267) of all CIN2 or above squamous epithelial lesions. Furthermore, three cases with pathological changes of the cervical severe glandular epithelium were all HPV18 infection. The difference was statistically significant (χ2=60.74, P<0.001). Single HPV subtype infection was primarily associated with HPV16, HPV52 and HPV58. In conclusion, HPV type detection had a may be important in screening of cervical lesions as a difference in pathogenic ability was noted among different HPV genotypes. As cervical cancer is an infectious disease, HPV testing may help detect more precancerous lesions, thus reducing the morbidity and mortality of cervical cancer. HPV16, HPV52 and HPV58 were associated with severe cervical squamous epithelial lesions; HPV18 was associated with cervical severe glandular cell pathological changes, although it was not the most common HPV genotype in China. When positive, a clinical cervical examination should be conducted, including colposcopy and biopsy.

Relationship between cyclin G1 and human papilloma virus infection in cervical intraepithelial neoplasia and cervical carcinoma.

OBJECTIVE: To evaluate the overexpression of cyclin G1 in cervical intraepithelial neoplasia (CIN) and cervical carcinoma, and the correlation between cyclin G1 and high-risk human papilloma virus (HPV) infection. METHODS: All of the specimens were obtained from the Department of Pathology of China-Japan Friendship Hospital from January 2000 to August 2004. We detected the expression of cyclin G1 with immunohistochemistry, HPV16/18 infection with in situ hybridization, and high-risk HPV infection with Hybrid capture system II (HC-II) in normal group (25 cases), CIN I (48 cases), CIN II (56 cases), CIN III (54 cases), and invasive cervical squamous-cell carcinoma (SCC, 31 cases). RESULTS: The positive rates of cyclin G1 expression in CIN (77.85%) and SCC cervical tissues (87.10%) were significantly higher than normal (8.00%, P < 0.01), and the intensities of cyclin G1 expression in CIN (40.60%) and SCC cervical tissues (61.51%) were significantly higher than normal (2.72%, P < 0.05). The positive rates and intensities of cyclin G1 expression increased gradually with the grade of cervical lesions. High-risk HPV infection rates were higher in CIN and SCC than normal groups (P < 0.05). There was a positive correlation between cyclin G1 expression and high-risk HPV infection detected with HC-II (Kendall's tau-b = 0.316, 0.269, 0.352, and 0.474 in CIN I, CINII, CIN III, and SCC, respectively, P < 0.05). CONCLUSIONS: Cyclin G1 is overexpressed in CIN and SCC. Cyclin G1 may be a biomarker for detecting CIN and SCC. Cyclin G1 may play an important role in the oncogenesis of CIN and SCC by high-risk HPV infection.

[Evaluation of clinical management strategies for atypical squamous cells of undetermined significance in cervical cytology].

OBJECTIVE: To evaluate the 3 different clinical management strategies for patients with cervical cytological atypical squamous cells of undetermined significance (ASC-US) recommended by the guideline of The 2001 Bethesda system and the 2001 American Society for Colposcopy and Cervical Pathology (ASCCP). METHODS: 1394 patients with a cytopathological diagnosis of ASC-US by use of liquid-based thin-layer preparation were managed by three different clinical strategies, and evaluated by the percentage of histological diagnosis > or = high-grade intraepithelial lesion (HSIL), i.e., cervical intraepithelial neoplasia (CIN2 and 3) as standard. 421 patients in Group A underwent colposcopically directed cervical biopsy, 475 patients in Group B were followed-up after 6 months by cytology, colposcopy and biopsy were performed if the results were > or = ASC-H or ASC-US and HPV-DNA (+). 498 patients in Group C: underwent HC-II test, colposcopy and biopsy were performed on those aged > or = 30 and with the HPV-DNA (+), if the patients were aged < 30 and with the HPV-DNA (+), HC-II test and cytology would be performed after 6 months; colposcopy and biopsy were performed on those with the results > or = ASC-H or HPV-DNA (+). RESULTS: (1) The results of histological diagnosis > or = CIN2 were found in 27 cases (6.41%) of Group A, 26 cases (5.78%) of Group B, and 34 cases (6.91%) of Group C. There was no statistically significant difference among these 3 groups (all P > 0.05). (2) Convenience was significantly different among these 3 groups (P < 0.01). The workloads for the doctors and the discomfort resulting from biopsy for the patients were the greatest in Group A. The patient's compliance of Group B was low because of the necessity to wait for follow-up six months later. The cost of Group C was relatively higher. (3) In Group C, 66% of the ASC-US patients with HPV-DNA (+) were aged > or = 30. The percentage of histological diagnosis > or = CIN2 was 5.69% in those aged > or = 30 and was 1.22% in those aged < 30. CONCLUSION: Both protocols B and C are practical for clinical management of ASC-US in China. HPV infection is the necessary cause of cervical carcinoma, so the protocol C (cytology and HC-II test) is better for cervical lesion screening. The ASC-US patients aged > or = 30 and with HPV-DNA (+) are at high risk.

[Application research on the flow-through hybridization and gene chip in human papillomavirus detection].

OBJECTIVE: To evaluate the clinical efficacy of flow-through hybridization and gene chip (HybriMax) on female low genital human papillomavirus (HPV) detection and to identify the most common HPV genotypes. METHODS: Five hundred and ninety-one of 7520 women who received cervical cancer screening in China-Japan Friendship Hospital during Jun 2004 to May 2005 were selected for 21 genotypes HPV detecting by HybriMax, including 138 women with normal cytological diagnosis and 453 women with abnormal cytological diagnosis. The abnormal patients were classified into groups according to their histological diagnosis as chronic cervicitis, cervical intraepithelial neoplasia (CIN) I, II, III, cervical cancer and condyloma acuminata. Among them 413 women were tested with HC-II for 13 types of high-risk HPV, and 101 paraffin-embedded specimens from these 453 patients were detected for HPV16, 18 with in situ hybridization (ISH). The accordance of the results was compared. The infection rates of each HPV type for each group were calculated. RESULTS: The total accordant rate of HybriMax and HC-II on 13 types of high-risk HPV detecting was 92.5% (382/413) and was 100.0% in group of cancer. The accordance was good, and the Kappa index (KI) was as below: overall 0.814, in normal group 0.750, in chronic cervicitis 0.781, in CIN I 0.755, in CIN II 0.619, in CIN III 0.548, in condyloma acuminata 0.800. The accordant rate of HybriMax and ISH was 89.1% (90/101), and KI was 0.766. The 10 most common genotypes were (in descending sequences): in normal group 16, 68, 18, 52, 58, 11, 53, 31, 39, 33; in abnormal group 16, 52, 58, 18, 33, 31, 81, 53, 68, 66, in cervical cancer 16, 18, 52, 58, 33, 66, 68, 31, 51, 53. CONCLUSIONS: HybriMax has good accordance to HC-II and ISH for HPV detection. It provides useful information on viral genotype and is more suitable for clinical use. The 6 most common genotypes in abnormal cytological group are 16, 18, 52, 58, 33, 31.

Cervical cytological screening and management in pregnant and postpartum women.

OBJECTIVE: To examine and follow up cervical cytology of pregnant and postpartum women and study their cytopathologic characteristics, so as to determine screening and managing programs for abnormal cervical cytology. METHODS: Totally 5296 patients in pregnancy and postpartum, in which 3729 by computer-assisted cytology test and 1567 by liquid-based monolayers cytology test, were examined and diagnosed by the Bethesda System made in 2001. Those proven epithelial abnormalities patients were followed up until the lesions regressed to normal. The remaining patients, who exhibited persistent abnormalities or progression, were given further examinations. RESULTS: The positive rate of cervical cytological test was 8.72% (462/5296), including squamous carcinoma (SCA) 1 case (0.02%), high grade squamous intraepithelial lesion (HSIL) 34 cases (0.64%), low grade squamous intraepithelial lesion (LSIL) 145 cases (2.74%), atypical glandular cells (AGC) 5 cases (0.09%), atypical squamous cells cannot exclude HSIL (ASC-H) 14 cases (0.26%), atypical squamous cells of undetermined significance (ASC-US) 263 cases (4.97%). The 419 proven cytological abnormality cases were followed up successfully. The total transnegative rate in three months was 73.74% (309/419), in which 303 cases (72.32%) persisted normal status for more than six months after regression. And the transnegative rate of ASC-US, ASC-H, AGC, LSIL, and HSIL were 79.56%, 64.29%, 100%, 72.14% and 44.12%, respectively. Forty-six cases received biopsy directed by colposcopy. The distribution of coincidence of cytopathologic and histopathologic diagnosis was: SCA 1 case (100%), HSIL 10 cases (76.92%), LSIL 13 cases (65%), ASC-H 2 cases (50%), ASC-US 3 cases (37.50%), total 29 cases (63.04%). CONCLUSIONS: We should cast more attention to screening cervix lesions in pregnant and postpartum women. Their cytopathologic characteristics are liable to make the clinician give a false positive diagnosis. So we propose to follow up them closely and to lower the indication of biopsy.

[Detection of integration status of human papillomavirus 16 in cervical precancerous lesions].

OBJECTIVE: To investigate the prevalence of integration of human papillomavirus 16 (HPV16) DNA into the host genome in cervical squamous intraepithelial lesions (SIL). METHODS: Multiplex PCR was used to detect the HPV/HPV16 infection and integration status of HPV16 in the surplus cells from liquid-based cytological samples from 108 patients with cervical cancer precursor lesions. Consensus primers GP5+/GP6+ were used to amplify a 150 bp long fragment in the conserved region of the HPV L1 gene so as to detect the presence pf HPV. Scion Image 4.0 electrophoresis image analysis soft was used to calculate the E2/E6 ratio so as to evaluate the episomal and integrated status of HPV16 infection: in episomal form, both targets should be equivalent, and in integrated form, E2 gene would be absent, while in mixed form of episomal/integrated mixed form, the copy number of E2 would be less than that of E6. RESULTS: Sixty-two out of the 108 patients (57.41%) had HPV infection. HPV16 were found in 32 of the 108 samples (29.63%). Among the 32 cases HPV16 DNA was exclusively episomal in 15 cases (46.88%), concomitant in 13 cases (40.62%), and integrated in 4 cases (12.50%). The prevalence of integrated and/or concomitant forms of HPV-16 DNA increased with progression of cervical disease. The prevalence of integrated form was 54.55% in the patients of CIN3 type, 50.00% in CIN2 type, 28.07% in CIN1 type, and 11.54% in the inflammatory type with significant differences between any 2 groups (all P < 0.01). CONCLUSIONS: HPV16 integration into the host genome is already present in some of CIN lesions. The multiplex PCR estimation of the HPV L1, HPV16 E2, E6 genes and E2/E6 ratio could be a simple method for detecting HPV/HPV16 infection and its integration status in liquid-based residual samples. It would be a helpful complementary tool for cytological screening to identify those patients at high risk of developing high-grade squamous intraepithelial lesions and cervical cancer.

[Comment on diagnosis of atypical squamous cells using new the Bethesda system 2001].

OBJECTIVE: To evaluate the criteria for diagnosis of atypical squamous cells (ASC) and its two subtypes: undetermined significance ASC (ASC-US) and atypical squamous cells-cannot exclude high-grade intraepithelial lesion (HSIL, ASC-H) defined in 2001 the Bethesda system (TBS). METHOD: One thousand one hundred and eighty six pap smears and liquid-based thin-layer preparations with diagnosis of atypical squamous cells of undetermined signification (ASCUS) according to TBS 1991 criteria, were re-classified by a cytopathologist using TBS 2001 criteria. RESULTS: The cytologic diagnosis according to TBS 2001 criteria was: negative in 285 cases (24.0 %), ASC-US in 627 cases (52.9%), ASC-H in 254 cases (21.4%), low-grade intraepithelial lesion (LISL) in 16 cases (1.3%), and HISL in 4 cases (0.3%), respectively. Out of 1186 cases, 168 received histological examination. Cervical intra-epithelial neoplasia (CIN) II-III were diagnosed in 18.9 % of patients with ASC-H, 5.2% of patients with ASC-US, and 14.3% of patients with ASCUS defined by using TBS 1991 criteria, respectively (P < 0.05). During the follow-up of 6 months, 4 cases of ASC-US changed to LSIL, one ASC-US changed to HSIL, 2 ASC-US changed to LSIL, and 2 ASC-US to HSIL. CONCLUSIONS: New classification of ASC recommended by 2001 TBS has apparent clinical significance. Colposcopically directed cervical biopsy should be performed immediately if ASC-H is diagnosed. In ASC-US patients, checking of cervical smear in an interval of 3 approximately 6 months is necessary. It is suggested to use ASC-US and ASC-H instead of ASCUS in cytological diagnosis.