Diagnostic value of the neutrophil-to-lymphocyte ratio and the combination of serum CA-125 for stages III and IV endometriosis.

BACKGROUND: Currently, all the diagnostic indicators for endometriosis lack perfect sensitivity and specificity. According to the characteristic of endometriosis, we analyzed the new biomarker neutrophil-to-lymphocyte ratio (NLR) and the combination of NLR and serum CA-125 to investigate their diagnostic value for identifying stages III and IV endometriosis. METHODS: The values of serum CA-125 and routine blood tests were collected from 197 patients with endometriosis, 102 with benign tumors and 112 healthy individuals. We investigated the sensitivity and specificity of NLR and its combination with serum-CA-125 for diagnosing stages III and IV endometriosis by using receiver operating characteristic (ROC). RESULTS: The mean values of NLR, the combination of serum CA-125 and NLR (combination) of the groups with stages III and IV endometriosis were significantly higher than the other two groups. Serum CA-125, NLR, and the combined biomarkers could significantly discriminate the stages III and IV endometriosis group from the other two groups (P < 0.05). NLR shows a lower sensitivity of 57.9% and specificity of 65.2% with a cutoff value at 1.82. And the combination of biomarkers has the highest AUC of 0.949 with a sensitivity of 86.8% and specificity of 92.0% at the cutoff value of 44.40. In addition, for patients with negative CA-125, 55.36% and 53.57% of the patients were able to be diagnosed with endometriosis by using NLR alone and the combination of biomarkers. CONCLUSION: For diagnosing stages III and IV endometriosis, the neutrophil-to-lymphocyte ratio is a better adjuvant to serum CA-125, and the neutrophil-to-lymphocyte ratio is valuable in diagnosing stages III and IV endometriosis for patients with negative serum CA-125.

Thymidylate synthase mRNA expression does not predict resistance to floxuridine in a choriocarcinoma cell line.

OBJECTIVE: Pyrimidine antagonist-based chemotherapy is a recommended chemotherapeutic treatment for gestational trophoblastic neoplasia in China. The main reason for treatment failure is chemoresistance. We established a floxuridine (FUDR)-resistant human choriocarcinoma JeG-3/FUDRA sub-line and investigated the role of thymidylate synthase (TS) transcript levels in chemoresistance prediction. STUDY DESIGN: The JeG-3/FUDRA sub-line was established by intermittent exposure to increasing doses of FUDR. Levels of TS transcripts were measured by real-time fluorescence quantitative reverse transcription-polymerase chain reaction. beta-hCG secretion in these cell lines has also been detected by using a chemoluminescence method. RESULTS: The JeG-3/FUDRA sub-line had a resistance index of 31.62. When exposed to 5.1 x 10(-7) M FUDR, the secretion of beta-hCG was enhanced dramatically, but with the increasing of the FUDR concentration, it was downregulated gradually. According to the concentration of FUDR exposure from low to high, the multiples of free beta-hCG secretion increased 13.19-, 4.76-, 1.90-, 1.44- and 0.47-fold, respectively. The TS transcript level was down-regulated by exposure to a low concentration of FUDR and then gradually increased with increasing doses of the drug. But when a certain concentration was reached, the expression would not increase but decrease sharply. By sub-lines of different concentration of FUDR exposure varying from low to high, the multiples of increases comparing the TS transcript level to parental cells were 0.56-, 0.42-, 1.02-, 2.78- and 2.06-fold, respectively. CONCLUSION: We successfully established an FUDR-resistant human choriocarcinoma sub-line and found that the expression of TS mRNA in FUDR-resistant JeG-3 cells was related to the concentration and phase of FUDR exposure. These data suggest that TS mRNA levels would be of limited use as biomarkers for the prediction of chemoresistance to FUDR-based chemotherapy.

[Establishment of floxuridine-resistant JeG-3 subline and the role of thymidylate synthetase mRNA expression in chem-resistant-prediction.].

OBJECTIVE: To establish human choriocarcinoma JeG-3 cell line resistant to floxuridine (FUDR) and describe the characteristics of this FUDR-resistant subline. The thymidylate synthase (TS) expression level in FUDR-resistant subline was also discussed. METHODS: The FUDR-resistant sub-line JeG-3/FUDRA was established by intermitted exposure to grads increased FUDR. Reversed microscope was used to observe the morphological changes in FUDR-resistant sub-line. Population doubling time was calculated and compared based on the growth curve of these two cell lines, cell cycles and chromosomal ploidy were assayed with flow cytometry methods. The chemo-luminescence assay was used to detect the hormone secretion by two kinds of cell lines. The resistant index (RI) was measured by cell counting kit-8 (CCK-8) assay. Quantitative RT-PCR was used to detect the mRNA expression level of TS and we also detected the TS mRNA expression level in different doses exposed subline. RESULTS: The RI of JeG-3/FUDRA was 31.62. Compared with the JeG-3 cell, the FUDR-resistant cell line had gross changes in morphological, cell growth, cell cycles and chromosomal numbers. The ability of human chorionic gonadotrop (hCG) and progesterone secretion was lower in JeG-3/FUDRA subline. The trend of TS mRNA expression was: while exposed to low concentration of FUDR, the TS mRNA expression level was downregulated, then followed the increasing dose of the drug, the expression level of TS mRNA ascended gradually. When the terminal concentration was reached, the expression level of TS mRNA in JeG-3/FUDRA subline was higher than that of JeG-3 cell line (P < 0.05). CONCLUSIONS: We established the FUDR-resistant subline of JeG-3 successfully. The TS mRNA expression level is stage-related to the different concentration and different phase in FUDR exposure. Our data suggested that TS mRNA expression level may not be used as a biomarker to predict the chemosensitivity in FUDR-based chemotherapy.

A potential mechanism of breakthrough bleeding associated with progestin: involvement in alteration of endometrial endothelial cells.

OBJECTIVE: To explore the potential mechanism of breakthrough bleeding associated with progestin with in vitro methods. METHODS: The isolation and culture of human endometrial endothelial cells (HEECs) was performed with the method established in our laboratory. The content and activity of urokinase-type plasminogen activator (uPA) and the content of plasminogen activator inhibitor-1 (PAI-1) in cell supernatants after incubated with different concentrations of progesterone (0-5 micromol/L) and 17beta-estradiol (0, 0.1, or 1 nmol/L) were measured by method of ELISA. Apoptosis rate of HEECs was measured by flow cytometry. Viable cell count was measured by MTT. RESULTS: The increased level of progesterone (0.5-5 micromol/L) combined with 17beta-estradiol elevated content and activity of uPA while the production of PAI-1 remained unchanged. The apoptosis of HEECs was inhibited along with the increment of total viable cell counts at higher concentrations of progesterone with 17beta-estradiol. CONCLUSION: The inhibition of apoptosis and increased content and activity of uPA may contribute to the occurrence of irregular bleeding associated with progestin use to some extent

Genistein inhibits proliferation of human endometrial endothelial cell in vitro.

OBJECTIVE: To explore the effect of genistein on proliferation of human endometrial endothelial cells (HEECs) and glandular epithelium. METHODS: In vitro HEECs and human endometrial cancer-1B cell (HEC-1B) were cultured with 0, 1, 10, 50, 100, and 200 micromol/L of genistein alone or indicated concentrations of genistein combined with 0.2 or 1 nmol/L 17beta-estradiol (17beta-E2). Cell proliferation was determined by [3H]-thymidine incorporation and cell cycle was measured by flow cytometry. RESULTS: After 96 hours of treatment, genistein inhibited the proliferation of HEECs in a dose-dependent manner. The stimulation index reduced from 100% (without genistein treatment) to about 1% (200 micromol/L genistein). HEECs were arrested at G1/0 and G2/M phase when treated with genistein for 96 hours. When the concentration of genistein was 200 micromol/L, the percentages of HEECs at G1/0, G2/M, and S phase were 96.0%, 2.1%, and 1.9%, respectively. However, when HEECs were treated without genistein, the percentages of HEECs at G1/0, G2/M, and S phase were 76.7%, 8.5%, and 14.7%, respectively. 17beta-E2 could not influence the effects of genistein on the proliferation of HEECs. Meanwhile, genistein could suppress the proliferation of HEC-1B. If the stimulation index of HEC-1B was defined as 100% when HEC-1B was treated with different doses of 17beta-E2 (without genistein), it was 67%, 19%, as well as 32% when cell was supplemented with 200 micromol/L genistein combined with 0, 0.2, or 1 nmol/L 17beta-E2, respectively. CONCLUSION: Genistein at the concentration of 200 micromol/L can sufficiently inhibit the proliferation of HEECs and endometrial glandular epithelium simultaneously in vitro.

[Incomplete P450 17 alpha enzyme deficiency: report of six cases].

OBJECTIVES: To summarize the characteristics, differential diagnosis and management of incomplete 17 alpha-hydroxylase/17, 20-lyase deficiency (17 OHD) of Chinese patients. METHODS: Six cases of incomplete 17 OHD from Peking Union Medical College Hospital were studied retrospectively through analyzing their clinical data, and the molecular pathogenic mechanism was discussed after literature review. RESULTS: Four cases of 46, XX incomplete 17 OHD were reported. The clinical characteristics included female phenotype, various degrees of breast development and absent or sparse axillary/pubic hair, oligomenorrhea or secondary amenorrhea, recurrent luteinized ovarian cysts, hypogonadism with persistent hyperprogesteronemia or high serum 17 alpha-hydroxyprogesterone level, with or without hypokalemic hypertension. There were also 2 cases of 46, XY incomplete 17 OHD, in which ambiguous genitalia were present besides hypokalemic hypertension. CONCLUSIONS: Incomplete 17 OHD is a very rare form of congenital enzymatic deficiencies of steroid synthesis, which should be included in the differential diagnosis when there are menstrual disorders, sexual infantilism, recurrent ovarian cysts or ambiguous genitalia. Under such circumstances, hyperprogesteronemia offers a valuable clue for further investigation.