Cytokines profiles in cervical mucosa in patients with cervical high-risk human papillomavirus infection.

INTRODUCTION: The purpose of this study was to detect the expression of local cytokines in cervical mucosa between patients with transient and persistent HR-HPV infection with or without CIN. METHODOLOGY: A total of 150 patients who were diagnosed as HR-HPV infection in Tianjin Central Hospital of Obstetrics and Gynecology from January 2016 to December 2016 were included in this study. The expression levels of 9 cytokines in 150 patients with HR-HPV infection, including interleukin (IL)-1ß, IL-4, IL-6, IL-8, IL-10, IL-12, IL-12p70, IL-21, interferon (IFN)-γ and tumor necrosis factor (TNF)-α, were simultaneously measured by using a multiplex immunoassay. Moreover, HR-HPV genotype was performed by using pyrosequencing. The association between cytokines and HPV genotype was also investigated. RESULTS: There was a statistically significant difference in IL-1ß level between patients with HPV transient infection and HPV persistent infection (p = 0.041). There were statistically significant differences in the levels of IL-1ß, IL-10, IL-21 and TNF-α between patients with low grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesion (HSIL) (p = 0.011, p = 0.008, p = 0.046 and p = 0.019, respectively). CONCLUSIONS: Pro-inflammatory cytokines, IL-1ß and TNF-α, and Th2 type cytokines, IL-10 and IL-21, became stronger in cervical mucosa with the progression of CIN. IL-1ß may be advantageous for HR-HPV persistent infection.

Value of circulating tumor cells positive for thyroid transcription factor-1 (TTF-1) to predict recurrence and survival rates for endometrial carcinoma.

PURPOSE: This study was conducted to analyze the predictive value of circulating tumor cells (CTC) expressing thyroid transcription factor-1 (TTF-1) on the recurrence and survival rates of endometrial carcinoma patients treated with laparoscopic surgery. METHODS: 78 patients were recruited, diagnosed with endometrial carcinoma and measured CTC expressing TTF-1 using flow cytometry in blood and tissues. Then, the patients were distributed into TTF-1-positive (N=42) and -negative (N=36) groups. The levels of HE4 were determined by ELISA, the levels of cancer antigen CA125 and CA15.3 by chemiluminescent immunoassay, and the levels of mRNA expression of survivin, ß-catenin, miR-15a, and PTEN by RT-PCR assay from endometrial carcinoma samples. RESULTS: Patients in TTF-1-positive group had mainly TNM stages II and III-IV, whereas the TTF-1-negative group stages I and II predominated. The rates of vascular infiltration and lymphatic metastasis in the TTF-1-positive group were higher compared with the TTF-1-negative group (p<0.05). The serum levels of CA125, CA15.3, and HE4 were significantly higher in the TTF-1-positive group than in the TTF-1-negative group (p<0.05). The levels of survivin and ß-catenin mRNA expression in endometrial carcinoma in the TTF-1-positive group was higher than in the TTF-1-negative group. In contrast, the levels of miR-15a and PTEN mRNA expression were lower in the TTF-1-positive group (p<0.05). The median follow-up duration was 25 months for both groups. At that time, progression-free survival (PFS) and the median survival time decreased in the TTF-1-positive group compared with the TTF-1-negative group. Additionally, the recurrence rate increased in the TTF-1-positive group. CONCLUSION: The rate of TTF-1-positive CTC was strongly correlated with TNM staging, vascular infiltration, lymphatic metastasis, and the levels of CA125, CA15.3, and HE4 in endometrial carcinoma. The levels of survivin, ß-catenin, miR-15a, and PTEN mRNA also contributed to predict survival rates after laparoscopic surgery.

Clinical experience of uterine tumors resembling ovarian sex cord tumors: a clinicopathological analysis of 6 cases.

OBJECTIVE: To compare the clinicopathological features, diagnosis, treatment, and prognosis of two types of uterine sex cord-like tumors. METHODS: The clinicopathological features of four uterine tumors resembling ovarian sex cord tumors (UTROSCTs) and two endometrial stromal tumors with sex cord-like elements (ESTSCLEs) were analyzed retrospectively. RESULTS: All patients were premenopausal women. The most common clinical presentation was vaginal bleeding (four cases). Total hysterectomy with or without bilateral adnexectomy was the most common treatment pattern (five cases). A patient with UTROSCTs, presenting with recurrence 10 months after transvaginal submucous myomectomy, underwent a total hysterectomy (case 2). All tumors were polypoid or intramural masses, usually located in the uterine fundus or submucosa. The majority of UTROSCTs were positive for cytokeratin (4/4 cases), one was positive for Wilms tumor protein, and of two cases with smooth muscle actin immunoreactivity, two were positive for desmin. UTROSCTs were positive for two or more sex cord markers, whereas sex cord markers were less frequently detected in ESTSCLEs. CD10 was variably positive in two UTROSCT patients and strongly positive in all ESTSCLE patients. Three UTROSCTs and one ESTSCLE were positive for both estrogen and progesterone receptors. All patients with UTROSCTs were alive without evidence of recurrence. One patient with ESTSCLEs underwent postoperative chemotherapy after total vaginal hysterectomy but developed recurrence at the vaginal stump (case 5). The other patient with ESTSCLEs was lost to follow-up. CONCLUSION: These UTROSCTs are polymorphic neoplasms with true sex cord differentiation and uncertain malignant potential, which possess a distinct biology from ESTSCLEs.

Establishing a Nomogram for Stage IA-IIB Cervical Cancer Patients after Complete Resection.

BACKGROUND: This study aimed to establish a nomogram by combining clinicopathologic factors with overall survival of stage IA-IIB cervical cancer patients after complete resection with pelvic lymphadenectomy. MATERIALS AND METHODS: This nomogram was based on a retrospective study on 1,563 stage IA-IIB cervical cancer patients who underwent complete resection and lymphadenectomy from 2002 to 2008. The nomogram was constructed based on multivariate analysis using Cox proportional hazard regression. The accuracy and discriminative ability of the nomogram were measured by concordance index (C-index) and calibration curve. RESULTS: Multivariate analysis identified lymph node metastasis (LNM), lymph-vascular space invasion (LVSI), stromal invasion, parametrial invasion, tumor diameter and histology as independent prognostic factors associated with cervical cancer survival. These factors were selected for construction of the nomogram. The C-index of the nomogram was 0.71 (95% CI, 0.65 to 0.77), and calibration of the nomogram showed good agreement between the 5-year predicted survival and the actual observation. CONCLUSIONS: We developed a nomogram predicting 5-year overall survival of surgically treated stage IA-IIB cervical cancer patients. More comprehensive information that is provided by this nomogram could provide further insight into personalized therapy selection.

A randomized study comparing the side effects and hormonal status of triptorelin and leuprorelin following conservative laparoscopic surgery for ovarian endometriosis in Chinese women.

OBJECTIVES: Different gonadotropin-releasing-hormone agonist (GnRH-a) formulations with different potency and associated side effects, therefore, different compliance and persistence of therapy. This study was to evaluate the difference of hormonal profile and side effects due to hypoestrogenic status after treatment of leuprorelin and triptorelin in Chinese women with ovarian endometrioma after conservative surgical treatment. STUDY DESIGN: A total of 302 women underwent laparoscopic excision of ovarian endometriomas with rASRM III and IV were enrolled in the study.Subjects were randomized into two groups with use of a random table. Twenty two patients dropped out during the study. Thus 142 patients had three doses of i.m. leuprorelin (group A) and 138 patients had three doses of i.m. triptorelin(group B) at 4 weeks intervals after surgical treatment. Menopausal symptoms were evalutaed using a questionnaire and serum sex hormonal levels were also measured during the follow-up. RESULTS: At week 4 after the treatment, most of the patients in leuprorelin group have no obvious side effects. After 9 weeks, bone pain, hot flashes and sweating, and irregular bleeding were the main side effects and showed no difference between the groups. Anxiety, depression, vaginal dryness, headache, and acne rates were all significantly higher in triptorelin group than in leuprorelin group. A significant difference in FSH (p=0.003), LH (p=0.026) and E2 (p=0.002) levels between the groups were observed after 21 days of the GnRHa treatment. The FSH (p=0.021) and E2 (p=0.033) levels remained higher in the leuprorelin group than the triptorelin group after six weeks of treatment, but the difference of LH(p=0.917) level was no longer discernible. CONCLUSION: Leuprorelin in down-regulating the pituitary-ovarian function was more moderate, and the hormonal levels decrease progressively and gradually, therefore, with lower rate of menopausal symptoms. Leuprorelin acetate maybe better tolerated than triptorelin.

[Relationship between the methylation of L1 3' and long control region gene of HPV16 DNA and it's pathogenicity].

OBJECTIVE: Quantifiably and located measure the methylation rate of 21 cytosine-phosphate-guanosine (CpG) sites in the 3' region of L1 gene and long control region (LCR) gene of HPV16 DNA in asymptomatic patients, cervical intraepithelial neoplasia (CIN) patients, and cervical cancer patients. To analysis the relationship between HPV16 methylation and it's pathogenicity. METHODS: Chosen 30 cases with HPV16 positive in each group. Firstly, extract DNA from the remaining cells of liquid-based cytology specimen and bisulfite treatment DNA, then amplify the 3' region of L1 gene and LCR gene, test the methylation rate of 21 CpG sites of HPV16 DNA in three groups. RESULTS: All of the 5 CpG sites in E6/E7 promoter (31, 37, 43, 52, 58) were hypomethylation in cervical cancer group (21.86%, 28.15%, 21.37%, 26.15%, 15.48%, respectively), hypermethylation in asymptomatic group, and middle-methylation in CIN group, in which there were significant difference among three groups (all P < 0.01). The CpG site in 7032, 7091, 7136 of the 3' region of L1 gene was also different methylated among three groups (all P < 0.01). Hypermethylation was found in cancer group (18.89%, 27.72%), hypomethylation was found in asymptomatic group (2.71%, 6.95%) in 7032 and 7091. In 7136, the highest methylation was detected in CIN (66.45%), the lowest in asymptomatic (34.85%), middle in cancer group (46.43%). CONCLUSION: The methylation status of CpG sites in the 3' region of L1 gene and E6/E7 promoter of HPV16 is significant different among three groups, which is likely to anticipate the pathogenesis of CIN and cervical cancer.

[The high expression of HPV31 and 52 L2 fusion protein in E. coli and detection of its immunogenicity].

OBJECTIVE: To express HPV31 and 52 L2 fusion protein and detect its immunogenicity. METHODS: According to the amino acid sequences of HPV31 and 52 L2 11-200AA published in the GenBank database, weartificially synthesized the HPV31 and 52 L2 fusion gene which was optimized according to Escherichia coli codon usage and encodes 11-200 amino acid of HPV31 and HPV52 L2, then cloned it into pET-9a vector. The HPV31 and 52 L2 fusion protein was expressed in Prokaryotic expression system and the mice were immunized with the fusion protein after purification. The immunogenicity was characterized in vaccinated mice. RESULTS: HPV31 and 52 L2 fusion protein was highly expressed in E. coli, the amount of fusion protein is nearly 20% of the total bacterial protein. The purified fusion protein with aluminum adjuvant could induce specific high titer of IgG antibodies detected by ELISA, and also induce the neutralizing antibodies against pseudovirus of HPV31 and HPV52 and cross-neutralizing antibodies against pseudovirus of HPV45, 58, 16, 18. CONCLUSION: HPV31 and 52 L2 fusion protein could induce neutralizing and cross-neutralizing antibodies against HPV pseudovirus. It provides laboratory basis for development of HPV L2 protein vaccine.

Clinical characteristics, metabolic features, and phenotype of Chinese women with polycystic ovary syndrome: a large-scale case-control study.

AIM: The Rotterdam criteria extend the phenotypic spectrum of polycystic ovary syndrome (PCOS). The study was to investigate the clinical and biochemical features of a large-scale clinic based on the samples of Chinese women and to evaluate the value of Rotterdam criteria on Chinese PCOS women. METHODS: One thousand four hundred and four Chinese women were involved in our study, among whom, 719 cases were diagnosed as PCOS based on 2003 Rotterdam criteria, and 685 women without history of hyperandrogenism and with regular menstrual cycles were recruited as control. Clinical features, ultrasonographic (ovarian follicle number and volume), hormonal and metabolic parameters were commenced as outcome measures. RESULTS: Among 719 PCOS women, 6.1 % had hirsutism, 13.3 % had acne, 21.1 % had hyperandrogenism, 94.2 % had polycystic ovaries on ultrasonographic examination, and 88.6 % had menstrual abnormality. About one-third of the total PCOS patients were insulin resistant. The most frequent PCOS phenotype is the non-hyperandrogenic phenotype (O + P). Total testosterone, LH/FSH ratio, body mass index (BMI), and Ferriman and Gallwey scores (F-G) were all significantly higher in PCOS groups compared with non-PCOS group. Women with PCOS and obesity had higher serum testosterone, fasting insulin, longer menstrual cycle and larger ovarian follicle number, and LH/FSH ratio, estradiol or ovarian volume were similar between obese and normal BMI women. The LH level was statistically lower in the obese PCOS group. CONCLUSIONS: Rotterdam criteria are generally applicable to Chinese population. Chinese women with PCOS showed lower rates of hyperandrogenemia, hirsutism, obesity, and insulin resistance. Obesity aggravates menstrual irregularity and increases the follicle number and serum total testosterone level.

Clinical outcomes of fertility-sparing treatments in young patients with epithelial ovarian carcinoma.

OBJECTIVE: To assess the clinical outcomes of fertility-sparing treatments in young patients with epithelial ovarian carcinoma (EOC). METHODS: A retrospective study of young EOC inpatients (≤40 years old) was performed during January 1994 and December 2010 in eight institutions. RESULTS: Data were analyzed from 94 patients treated with fertility-sparing surgery with a median follow-up time of 58.7 months. As histologic grade increased, overall survival (OS) and disease-free survival (DFS) of patients receiving fertility-sparing surgery declined. Neither staging surgery nor laparoscopy of early stage EOC with conservative surgery had a significant effect on OS or DFS. Normal menstruation recommenced after chemotherapy in 89% of the fertility-sparing group. Seventeen pregnancies among twelve patients were achieved by the end of the follow-ups. CONCLUSIONS: Fertility-sparing treatment for patients with EOC Stage I Grade 1 could be cautiously considered for young patients. The surgical procedure and surgical route might not significantly influence the prognosis. Standard chemotherapy is not likely to have an evident impact on ovarian function or fertility in young patients.

[E-cadherin promoter methylation and demethylation in epithelial ovarian carcinoma cells].

OBJECTIVE: To investigate the cytidylyl phosphate guanosine (CpG) islands methylation status of E-cadherin (E-cad) promoter region in human ovarian carcinoma cell lines (ES-2,3AO,SKOV3), and the effect of 5-azacytidine-2'-deoxycytidines (5-Aza-CdR) on the cell proliferative ability, invasion and the expression of E-cad protein. METHODS: Methylation specific PCR (MSP) was used to detect CpG islands methylation status of E-cad promoter region in ES-2,3AO and SKOV3 cell lines. After treated with different concentrations of 5-Aza-CdR, morphological changes of cell lines were observed under microscope. The proliferative ability was evaluated by methyl thiazolyl tetrazolium (MTT) assay. E-cad protein expression was detected by western-blot and cellular invasion was investigated by 24-well matrigel invasion chambers. RESULTS: Hypermethylation status of CpG islands of E-cad promoter region was observed in ES-2 and SKOV3 cell lines, but not in 3AO cell lines. After treated with 5-Aza-CdR (0.1, 1.0, 10.0 micromol/L), ES-2 and SKOV3 cell lines displayed morphological evidence of differentiation. 5-Aza-CdR was found to decrease proliferation as evidenced by cell growth curve , to increase the level of E-cad protein expression (P < 0.01), and effectively inhibit the ability of cell invasion (P < 0.01). CONCLUSIONS: CpG hypermethylation is an important mechanism of E-cad gene inactivation in ES-2 and SKOV3 cell lines. 5-Aza-CdR be found to inhibit proliferation and invasion, and increase the expression of E-cad probably by the inhibition of hypermethylation.