Extended HPV Genotyping for Risk Assessment of Cervical Intraepithelial Neoplasia Grade 2/3 or Worse in a Cohort Study.

BACKGROUND: We sought to identify the absolute risk of specific HPV genotype for cervical intraepithelial neoplasia grade 2/3 or worse (CIN2+/3+) and to develop a risk-based management strategy in an HPV-positive population. METHODS: HPV genotyping was performed based on a 3-year cervical cancer screening cohort. The study endpoints were histologic CIN2+/3+. The prevalence of specific HPV genotype was calculated by minimum, any type, and hierarchical attribution estimate. The absolute CIN2+/3+ risks of specific HPV genotype were estimated and risk-based management strategy was established according to the American Society for Colposcopy and Cervical Pathology guideline. The efficacy of conventional and risk-based management strategies for non-16/18 HPVs were further evaluated. RESULTS: Eligible data were available for 8,370 women with a median age of 48 years (interquartile range, 42-53 years). At baseline, there were 1,062 women with HPV-positive disease, including 424 with multiple and 639 with single infections. CIN2+/3+ cases represented 113/74, 23/8, 20/7, and 52/31 patients at baseline and first-, second-, and third-year visits, respectively. Women with multiple HPV infections at baseline were more prone to persistent infection than those with single infection (P<.0001). HPV16 and HPV52 were the top 2 ranking among baseline and 3-year cumulative CIN2+/3+ cases. Based on the absolute risk of specific HPV genotype combined with cytology for CIN2+/3+, all non-16/18 HPVs were divided into 4 risk-stratified groups. Compared with conventional strategy, the risk-based strategy had higher specificity (P=.0000) and positive predictive value (P=.0322) to detect CIN3+ and needed fewer colposcopies for each CIN3+ case. CONCLUSIONS: Based on our study findings, we propose a new extended HPV genotyping protocol, which would provide a better strategy for achieving precise risk-based management of HPV-positive populations.

MiRNA detection in cervical exfoliated cells for missed high-grade lesions in women with LSIL/CIN1 diagnosis after colposcopy-guided biopsy.

BACKGROUND: Low-grade squamous intraepithelial lesion/cervical intraepithelial neoplasia grade 1 (LSIL/CIN1) preceded by colposcopy guided biopsy is recommended conservative follow-up, although some of these lesions are actually high-grade lesions, which are missed on an initial colposcopy. Therefore, in this work, we evaluate the potential role of miRNA detection in cervical exfoliated cells in a clinic-based population for predicting missed high-grade lesions in women diagnosed with LSIL/CIN1 after colposcopy-guided biopsy. METHODS: A total number of 177 women with a diagnosis of LSIL/CIN1 obtained by colposcopy-guided biopsy were grouped into two categories according to the histology of the conization specimens: consistent LSIL/CIN1 group (surgical pathology consistent with colposcopic diagnosis) and missed high-grade lesion group (surgical pathology found high-grade lesion). The expression of eight miRNAs, such as miRNA195, miRNA424, miRNA375, miRNA218, miRNA34a, miRNA29a, miRNA16-2, and miRNA20a was detected by real time-quantitative polymerase chain reaction (RT-qPCR) in cervical exfoliated cells of the 177 patients. Pearson Chi-Square was used to compare the performance efficiency of patients' characteristics. Nonparametric Man-Whitney U test was used to assess differences in miRNA expression. The receiver operating characteristic (ROC) curve was used to assess the performance of miRNA evaluation in detecting missed high-grade lesions. RESULTS: Among the 177 women with biopsy-confirmed CIN1, 15.3% (27/177) had CIN2+ in the conization specimen (missed high-grade lesion group) and 84.7% (150/177) had CIN1-(consistent LSIL/CIN1 group). The relative expression of miRNA-195 and miRNA-29a in the missed high-grade lesion group was significantly lower than that in the consistent LSIL/CIN1 group. The relative expression of miRNA16-2 and miRNA20a in the missed high-grade lesion group was significantly higher than that in the consistent LSIL/CIN1 group. No significant difference was observed between these two groups regarding the other four miRNAs. Of these significant miRNAs, miRNA29a detection achieved the highest Youden index (0.733), sensitivity (92.6%), positive predictive value (46.2%), negative predictive value (98.3%) and higher specificity (80.7%) when identifying missed high-grade lesions. CONCLUSIONS: Detection of miRNA might provide a new triage for identifying a group at higher risk of missed high-grade lesions in women with colposcopy diagnosis of LSIL/CIN1.

The inhibition of UBC13 expression and blockage of the DNMT1-CHFR-Aurora A pathway contribute to paclitaxel resistance in ovarian cancer.

Paclitaxel is widely used as a first-line chemotherapeutic drug for patients with ovarian cancer and other solid cancers, but drug resistance occurs frequently, resulting in ovarian cancer still presenting as the highest lethality among all gynecological tumors. Here, using DIGE quantitative proteomics, we identified UBC13 as down-regulated in paclitaxel-resistant ovarian cancer cells, and it was further revealed by immunohistochemical staining that UBC13 low-expression was associated with poorer prognosis and shorter survival of the patients. Through gene function experiments, we found that paclitaxel exposure induced UBC13 down-regulation, and the enforced change in UBC13 expression altered the sensitivity to paclitaxel. Meanwhile, the reduction of UBC13 increased DNMT1 levels by attenuating its ubiquitination, and the up-regulated DNMT1 enhanced the CHFR promoter DNA methylation levels, leading to a reduction of CHFR expression, and an increased in the levels of Aurora A. Our findings revealed a novel function for UBC13 in regulating paclitaxel sensitivity through a DNMT1-CHFR-Aurora A pathway in ovarian cancer cells. UBC13 could potentially be employed as a therapeutic molecular drug for reversing paclitaxel resistance in ovarian cancer patients.

Prognostic value of human papillomavirus 16/18 genotyping in low-grade cervical lesions preceded by mildly abnormal cytology.

Histological low-grade squamous intraepithelial lesion/cervical intraepithelial neoplasia grade 1 (LSIL/CIN1) preceded by normal or mildly abnormal cytology is recommended for conservative follow-up, with no separated management. In this study, we assessed the triage value of human papillomavirus (HPV) 16/18 genotyping in 273 patients with LSIL/CIN1. HPV16/18 genotyping was performed at baseline and follow-up was at 6-monthly intervals for up to 2 years. At each follow-up, women positive for cytology or high-risk HPV (hrHPV) were referred for colposcopy. Enrollment cytology, HPV16/18 genotyping, and questionnaire-obtained factors were linked to the 2-year cumulative progression rate. Univariate and multivariate analyses were performed taking into account time-to-event with Cox proportional hazard regression. The results showed that 190 cases (69.6%) regressed, 37 (13.6%) persisted, and 46 (16.8%) progressed. HPV16/18 positivity (hazard ratio (HR), 2.708; 95% confidence interval (CI), 1.432-5.121; P=0.002) is significantly associated with higher 2-year cumulative progression rate. Sub-analysis by enrollment cytology and age restricted the positive association among patients preceded by mildly abnormal cytology and aged 30 years or older. Immediate treatment is a rational recommendation for the high-risk subgroup, when good compliance is not assured.

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

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

TXNDC17 promotes paclitaxel resistance via inducing autophagy in ovarian cancer.

Paclitaxel is recommended as a first-line chemotherapeutic agent against ovarian cancer, but drug resistance becomes a major limitation of its success clinically. The key molecule or mechanism associated with paclitaxel resistance in ovarian cancer still remains unclear. Here, we showed that TXNDC17 screened from 356 differentially expressed proteins by LC-MS/MS label-free quantitative proteomics was more highly expressed in paclitaxel-resistant ovarian cancer cells and tissues, and the high expression of TXNDC17 was associated with poorer prognostic factors and exhibited shortened survival in 157 ovarian cancer patients. Moreover, paclitaxel exposure induced upregulation of TXNDC17 and BECN1 expression, increase of autophagosome formation, and autophagic flux that conferred cytoprotection for ovarian cancer cells from paclitaxel. TXNDC17 inhibition by siRNA or enforced overexpression by a pcDNA3.1(+)-TXNDC17 plasmid correspondingly decreased or increased the autophagy response and paclitaxel resistance. Additionally, the downregulation of BECN1 by siRNA attenuated the activation of autophagy and cytoprotection from paclitaxel induced by TXNDC17 overexpression in ovarian cancer cells. Thus, our findings suggest that TXNDC17, through participation of BECN1, induces autophagy and consequently results in paclitaxel resistance in ovarian cancer. TXNDC17 may be a potential predictor or target in ovarian cancer therapeutics.

Acid-sensing ion channel 1a is involved in acid-induced osteoclastogenesis by regulating activation of the transcription factor NFATc1.

It has been known that osteoclastogenesis is induced by extracellular acidosis-evoked the rise of intracellular calcium ([Ca(2+)]i), which regulate activation of the transcription factor nuclear factor of activated T cells c1 (NFATc1). However, the acid-sensing ion channels (ASICs) involved remain largely unknown. Here, we show that ASIC1a, ASIC1b, ASIC2a, and ASIC3 are expressed in rat osteoclasts, and only ASIC1a is highly upregulated in response to acidosis. Both the ASIC1a-specific blocker PcTX1 and specific siRNA significantly reduce this increase in acid-induced [Ca(2+)]i and acid-induced nuclear translocation of NFATc1, and inhibit acid-induced osteoclast differentiation and bone resorption. These findings show that ASIC1a-mediated calcium entry plays a critical role in osteoclastogenesis by regulating activation of the NFATc1.

[Risk factors of predicting residual disease in women with stage I a1 squamous cervical carcinoma after conization].

OBJECTIVE: To evaluate the prevalence and influencing factors of residual disease in women with stage I a1 squamous cervical carcinoma after conization. METHODS: The medical records and histopathologic slides of 83 women diagnosed with stage I a1 squamous cervical carcinoma after cervical conization undergoing subsequent hysterectomy at our hospital between January 2003 and December 2007 were reviewed. The correlations between the presence of residual lesions and clinicopathological features were analyzed. RESULTS: Among them, 31 (37.3%) had residual disease in hysterectomy specimens, including CIN1 (n = 5), CIN2-3 (n = 10), microinvasive carcinoma (n = 11) and invasive carcinoma (n = 5). In univariate analysis, menopause, procedure of conization, and status of cone margins were associated with the prevalence of residual disease in stage I a1 cervical carcinoma after conization. However, Logistic regression analysis revealed status of cone margins as an independent risk factor for residual disease in stage I a1 cervical carcinoma after conization. CONCLUSION: Status of cone margins is an independent risk factor for residual disease in stage I a1 cervical carcinoma after conization. Further treatment should be performed in patients with positive or nearing cone margins.

Effect of leonurine hydrochloride on endothelin and the endothelin receptor-mediated signal pathway in medically-induced incomplete abortion in rats.

OBJECTIVE: Endothelin (ET) is involved in uterine contractions. Our previous study showed that leonurine hydrochloride (LH) inhibits abnormal bleeding caused by incomplete abortion through an increase in uterine contractions in rats. The present study was conducted to show that LH treatment regulates the ET-mediated signal pathway in abortion in rats. STUDY DESIGN: Early pregnancies in rats had incomplete abortions induced using mifepristone in combination with misoprostol. After the abortions, the rats were treated with LH orally for 7 days and surgery was performed. The sinistro-uterus was dissected for measurement of ET and nitric oxide (NO); the dextro-uterus was stored at -80°C for ET receptor (ETA and ETB) analysis. Myometrial cells from the dextro-uterus were cultured for measurement of phospholipase C (PLC) activity, intra-cellular Ca(2+) concentration ([Ca(2+)]i), and protein kinase C (PKC) activity. RESULTS: In in vivo experiments, LH treatment elevated the ET level and ET/NO ratio in rats with induced abortions and up-regulated ETA mRNA expression (P<0.01 vs. the model group), but there was no change in ETB mRNA. LH significantly increased the [Ca(2+)]i, PLC activity, and relative production of PKC protein in myometrial cells. CONCLUSION: LH increased uterine contractions in rats with incomplete abortions by modulating the ET receptor-mediated signal pathway.

Epidermal growth factor receptor (EGFR) as a therapeutic target in rheumatoid arthritis.

The epidermal growth factor receptor (EGFR) has an important role in the hyperplastic growth of tumor. Similar to tumor growth, rheumatoid arthritis (RA) synovium is hyperplastic, invasive, and expresses EGFR and its ligands. Activation of EGFR signaling is responsible for synovial fibroblast proliferation in RA. Furthermore, in addition to its role in proliferation, EGFR and its ligands can induce cytokine production of synovial fibroblasts during the pathogenesis of RA. Agents that target EGFR have yielded promising results in animal experiments involving RA, pharmacologic modulations targeting EGFR, or its ligands may give rise to new therapeutic approaches for RA. In this review article, we will discuss the biological features of EGFR and summarize recent advances regarding the role of EGFR in the pathogenesis and treatment of RA.

[Reversing paclitaxel-resistance of SKOV3-TR30 cell line by curcumin].

OBJECTIVE: To explore the effects of curcumin on paclitaxel resistance reversal of SKOV3-TR30 cell line and its mechanism. METHODS: The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) MTT assay was performed to determine the sensitivity of curcumin-treated SKOV3-TR30 cells to paclitaxel. The cell cycle distribution of SKOV3-TR30 was analyzed by flow cytometry. And the expression level of glycogen synthase kinase-3 (GSK-3) protein was detected by Western blot. RESULTS: IC(50) of paclitaxel in SKOV3-TR30 decreased with a treatment of curcumin. And the reversal times was 3.0. Flow cytometric analysis of curcumin-treated SKOV3-TR30 cells demonstrated a distinct G(2)-M phase block (78.5 ± 6.4)% after a 12-hour treatment of paclitaxel versus SKOV3-TR30 cells without curcumin. There was a lack of G(2)-M phase arrest (only 27.0% ± 2.9%). The expression of GSK-3 protein in SKOV3-TR30 cells decreased with the 12 and 24-hour treatments of curcumin. CONCLUSION: Curcumin can partially reverse the paclitaxel-resistance of SKOV3-TR30 cells through a down-regulation of GSK-3.

Type 17 T-helper cells might be a promising therapeutic target for osteoporosis.

Osteoporosis, a disease characterized by low bone mass and deterioration of bone tissue, is a pressing public health problem. Recent studies have suggested a possible role of T-helper (Th) cells in the pathogenesis of bone loss which occurs in systemic inflammatory diseases. However, there are contradictions in the published literature regarding the functional role of Th1/Th2 cells in the regulation of the differentiation of osteoclasts. These paradoxes have now been clarified by the recent discovery of Th17 cells, a novel subset of Th cells that selectively secrete several proinflammatory cytokines, mainly IL-17. It has been confirmed that Th17 cells have stimulatory effects on osteoclastogenesis and accelerate bone loss in animal models with inflammatory disorders. Targeting Th17 cells or IL-17 may inhibit the bone resorption with RA. Thus, we are led to suppose that Th17 cells might be promising therapeutic targets in osteoporosis.

[Expression and significance of LMP2 and PPM1A in gestational trophoblastic disease].

OBJECTIVE: To investigate the expression of low molecular mass polypeptide-2 (LMP2) and protein phosphatase 1A (PPM1A) in gestational trophoblastic disease and elucidate their predictive value in malignant transformation of hydatidiform mole. METHODS: The expressions of LMP2 and PPM1A protein in 196 complete hydatidiform moles (in which 28 cases with malignant transformation), 7 invasive moles, 5 choriocarcinomas and 20 normal chorionic villus were detected with the method of EnVision immunohistochemistry. Their clinicopathologic data were retrospectively analyzed. RESULTS: LMP2 and PPM1A protein expressed in cytotrophocytes, syncytiotrophoblast and extravillous trophoblast. The level of LMP2 expression in deteriorative hydatidiform mole was significantly higher than that in non-deteriorative hydatidiform mole or normal chorionic villus (6.79±2.38, 5.26±2.63 and 3.10±1.65, all P<0.01), while there were no difference compared with gestational trophoblastic neoplasms (6.42±2.68, P=0.113). The level of PPM1A expression was highest in normal chorionic villus, and decreased gradually in hydatidiform mole (non-deteriorative and deteriorative) and gestational trophoblastic neoplasms (6.30±2.98, 4.93±2.50, 4.43±2.04 and 3.33±2.06, all P<0.01); the level of PPM1A expression in deteriorative hydatidiform mole was significantly lower than that in non-deteriorative hydatidiform mole (P=0.001). The expression of LMP2 protein was correlated to theca lutein ovarian cyst, the expression of PPM1A protein was related with uterine size (P<0.05). While, there was no correlation between the expressions of the two proteins (P>0.05). CONCLUSIONS: High expression of LMP2 and low expression of PPM1A might play an important role in the motility and invasiveness of trophoblast cells and malignant transformation of hydatidiform mole. Testing the expression of LMP2 and PPM1A in hydatidiform mole tissues of initial uterine evacuation might be have some reference significance in judging outcomes of hydatidiform mole.

Effects of leonurine hydrochloride on medically induced incomplete abortion in early pregnancy rats.

OBJECTIVES: To determine the effect of leonurine hydrochloride (LH) on abnormal bleeding induced by medical abortion. STUDY DESIGN: Rats had incomplete abortions induced in early pregnancy using mifepristone in combination with misoprostol. After abortion, rats were treated with LH for 7 days, and the duration and volume of uterine bleeding were observed. Approximately 30min after the last treatment, the animals were killed and the uterine shape was observed. The sinistro-uteri were suspended in organ baths to record the contraction curves, including the frequency and tension for 10min; the dextro-uteri were fixed with formaldehyde for pathologic evaluation. In addition, blood samples were collected from the femoral artery for the measurement of estradiol (E2) and progesterone (P) levels by radioimmunoassay. RESULTS: In in vivo experiments, compared with the model group, LH treatment markedly reduced the volume of bleeding and intrauterine residual, and significantly shortened the duration of bleeding. From the contraction curve, LH notably reinforced the frequency and tension of uterine contractions. LH remarkably elevated the serum estradiol level in rats, but had no obvious effect on progesterone level. CONCLUSIONS: LH has an inhibitory effect on bleeding caused by incomplete abortion; the mechanism may be related to up-regulation of the E2 level, leading to an increase in uterine contractions and evacuation of intrauterine residuum.

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. P16(INK4) (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.

Reduced miR-100 expression in cervical cancer and precursors and its carcinogenic effect through targeting PLK1 protein.

AIM: Although aberrant miRNAs expression has been documented, altered miR-100 expression in cervical cancer and precursor tissues and its carcinogenic effect and mechanism remain unexplored. The aim of our study was to investigate the role of miR-100 alteration in cervical carcinogenesis. METHODS: The expression of miR-100 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in 125 cervical tissues including normal cervical epithelium, cervical intraepithelial neoplasia (CIN), and cervical cancer, as well as in five cervical cell lines. Through modulating miR-100 expression using miR-100 inhibitor or mimic in vitro, cell growth, cycle and apoptosis were tested separately by MTT or flow cytometry and meanwhile Polo-like kinase1 (PLK1) mRNA and protein expressions were detected by qRT-PCR and immunoblotting. The expression of PLK1 in 125 cervical tissues was also examined by immunohistochemical staining and the correlation between miR-100 and PLK1 expression in the same tissues was analysed. Finally, HPV-16 E6/E7 expression was modulated by gene transfection and subsequently the level of miR-100 was examined by qRT-PCR. RESULTS: The miR-100 expression showed a significantly and gradually reduced tendency from low-grade CIN, high-grade CIN to cervical cancer tissues and a significant decrease in HPV positive cervical cancer cell lines. The modulation of miR-100 expression remarkably influenced cell proliferation, cycle and apoptosis, as well as the level of PLK1 protein, but not mRNA, in vitro experiments. PLK1 expression was negatively correlated with miR-100 expression in CIN3 and cervical cancer tissues. The modulation of HR-HPV E6/E7 expression did not change miR-100 level. CONCLUSIONS: The reduced miR-100 expression participates in the development of cervical cancer at least partly through loss of inhibition to target gene PLK1, which probably occurs in a relative late phase of carcinogenesis. HR-HPV E6/E7 may not directly regulate miR-100 expression in cervical cells.

[Histological evaluation of cervical carcinomas in FIGO stage Ib2/IIa after neoadjuvant chemotherapy].

OBJECTIVE: To investigate the histological changes of cervical cancer after neoadjuvant chemotherapy (NACT) and to establish histological criteria for interpretation of chemotherapeutical effects. METHODS: Fifty-six patients with FIGO stage Ib2-IIa cervical cancers treated by NACT and subsequent radical surgery were retrospectively analyzed, in which the pre- and post-chemotherapeutic histopathological changes were assessed. RESULTS: The post-chemotherapeutic histopathological changes of 56 cases included grade 3 effects in 11 cases (19.6%), grade 2 in 24 cases (42.9%), grade 1 in 13 cases (23.2%) and no response in only 8 cases (14.3%). The histologic response rate was 62.5% (35/56) and the overall clinical response rate was 67.9% (38/56). The overall coincidence by both criteria was 78.6% (44/56). Four cases (7.1%, 4/56) had only histological response and 8 cases (14.3%, 8/56) had response by imaging. In comparison with the pre-chemotherapy specimens, the chemotherapy-associated histological changes included shrinkage and scattering of tumor nests,decrease of tumor cellularity,tumor cell degeneration and necrosis. CONCLUSIONS: The histological changes in locally advanced cervical cancers induced by NACT are significant, which may challenge the diagnosis in the final specimens. There are some discreqancies between the histological criteria and imaging/gynecological ones for the therapeutic evaluation of cervical cancers,and it is thus recommended to use the pathological criteria for clinic practice.

[Evaluation of five screening methods for an early detection of cervical cancer and its precancerous lesions in Zhejiang province].

OBJECTIVE: To evaluate five screening methods of cervical cancer so as to popularize an effective screening strategy for cervical cancer in Zhejiang province. METHODS: A total of 1005 women aged 25 - 65 years old were selected from Lishui where cervical cancer was highly prevalent. And 859 subjects were ultimately enrolled between June 2009 and December 2009. Each subject was subjected to five screening methods, including Pap smear, liquid-based cytology (LBC), human papillomavirus DNA with a second-generation hybridization assay (HC2), visual inspection with acetic acid (VIA) and visual inspection with Lugol's iodine (VILI). CIN (cervical intraepithelial neoplasia) 2+ on biopsy was used as the reference standard for disease positivity. Negative colposcopy was accepted as a negative outcome. RESULTS: The sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV) were 25%, 90%, 26.7% and 98.6% for Pap smear; 81.3%, 97.3%, 35.1% and 99.6% for LBC; 68.9%, 82.8%, 7.1% and 99.3% for VIA; 81.3%, 84.6%, 9.1% and 99.3% for VILI; 87.5%, 77.3% and 6.8% for HPV-DNA test respectively. CONCLUSIONS: LBC is associated with a better profile of sensitivity, specificity and predictive value for five screening methods. It has the potential of optimizing the effectiveness of primary cervical cancer screening. Due to a low cost and an easy operation, VIA screening is an effective method of screening cervical cancer in the underdeveloped areas.

Overexpression of glycogen synthase kinase-3 in ovarian carcinoma cells with acquired paclitaxel resistance.

INTRODUCTION: Acquired resistance to paclitaxel, including regimens, is one of the most significant reasons for treatment failure and death in patients with ovarian cancer, but the causes of this resistance remain unclear. However, cell cycle regulation is a key mechanism by which most chemotherapeutic agents exert their cytotoxic effects. METHODS: We created a paclitaxel-resistant ovarian carcinoma cell line from SKOV3 cell line, and the difference of cell cycle distribution was analyzed using flow cytometry. Analysis of human cell cycle pathway complementary DNA array was performed to identify candidate genes associated with paclitaxel resistance. Gene expression changes were validated at the messenger RNA and protein levels by real-time reverse transcriptase polymerase chain reaction and Western analysis, respectively. RESULTS: The ratio of Gap0/Gap1 phase in SKOV3-TR30 was significantly lower than that in SKOV3 (54.8% ± 6.3% vs 72.7% ± 7.6%, P = 0.035), and the ratio of G2/M phase in SKOV3-TR30 was significantly higher than that in SKOV3 (24.9% ± 6.0% vs 10.2% ± 3.5%, P = 0.021). Complementary DNA microarray analysis demonstrated enhanced glycogen synthase kinase-3α (GSK-3α) expression in paclitaxel-resistant ovarian carcinoma cells. Real-time reverse transcriptase polymerase chain reaction analysis revealed that the paclitaxel-resistant subline exhibited a 7.0 ± 1.8-fold increase in GSK-3α messenger RNA expression. There was a 3.34 ± 0.47-fold increase of total GSK-3 protein (GSK-3α/ß) in SKOV3-TR30 cells validated by Western analysis. CONCLUSIONS: This study demonstrates that enhanced expression of GSK-3 is associated with acquired resistance to paclitaxel in ovarian carcinoma cells. Glycogen synthase kinase-3 overexpression may probably be a significant contributor to chemoresistance.