A retrospective study of ovarian tissue cryopreservation in female patients with hematological diseases for fertility preservation.

PURPOSES: To investigate the effect and safety of ovarian tissue cryopreservation (OTC) for fertility preservation in female patients with hematological diseases. METHODS: We designed a retrospective study. The clinical data of patients with hematological diseases undergoing OTC admitted to Peking University People's Hospital from April 2017 to January 2023 were analyzed and summarized. RESULTS: A total of 24 patients were included in the study, including 19 patients with malignant hematological diseases and 5 patients with non-malignant hematological diseases. The former included 14 patients with acute leukemia, 1 patient with chronic leukemia, and 4 patients with myelodysplastic syndrome, while the latter 5 patients were aplastic anemia (AA). 16 patients had received chemotherapy before OTC. The average age of 24 patients was 22.80 ± 6.81 years. The average anti-Mullerian hormone (AMH) was 1.97 ± 2.12 ng/mL, and the average follicle-stimulating hormone (FSH) was 7.01 ± 4.24 IU/L in examination before OTC. FSH was greater than 10.0 IU/L in 4 cases. The pre-OTC laboratory tests showed that the average white blood cell (WBC) count was (3.33 ± 1.35) × 109/L, the average hemoglobin was 91.42 ± 22.84 g/L, and the average platelet was (147.38 ± 114.46) × 109/L. After injection of recombinant human granulocyte colony-stimulating factor (rhG-CSF), blood transfusion, and iron supplementation in pre-OTC treatment, the average WBC count was (4.91 ± 3.07) × 109/L, the average hemoglobin was 98.67 ± 15.43 g/L, and the average platelet was (156.38 ± 103.22) × 109/L. Of the 24 patients, 22 underwent laparoscopic bilateral partial oophorectomy and oophoroplasty, and 2 underwent laparoscopic unilateral oophorectomy. The average duration of OTC was 59.54 ± 17.58 min, and the average blood loss was 32.1 ± 41.6 mL. The maximum blood loss was 200 mL. There was no significant difference in WBC count and hemoglobin concentration after OTC compared to pre-OTC period. Only the platelet count after OTC surgery was significantly different from that before surgery ([134.54 ± 80.84 vs. 156.38 ± 103.22] × 109/L, p < 0.05). None of the 24 patients had serious complications after OTC. 2 patients had mild infection symptoms, but both recovered well. 23 patients underwent hematopoietic stem cell transplantation (HSCT) after OTC. The median and interquartile range from OTC to the pretreatment of HSCT was 33 (57) days, and the median and interquartile range from OTC to HSCT was 41 (57) days. Seven of them began pretreatment of HSCT within 20 days and began HSCT within 30 days after OTC. All patients were followed up. Of the 23 patients who underwent HSCT after surgery, 22 presented with amenorrhea and 1 with scanty menstrual episodes. Seven patients underwent hormone replacement therapy (HRT) after HSCT. A patient with AA underwent ovarian tissue transplantation (OTT) 3 years after HSCT and resumed regular menstruation 6 months after OTT. CONCLUSIONS: Ovarian tissue cryopreservation has a promising future in fertility protection in patients with hematological diseases. However, patients with hematological malignancies often have received gonadotoxic therapy before OTC, which may be accompanied by myelosuppression while patients with non-malignant hematological diseases often present with severe hemocytopenia. So perioperative complete blood count of patients should be paid attention to. There was no significant difference in the WBC count and hemoglobin concentration in patients with hematological diseases before and after OTC surgery, and the platelet count decreased slightly within the normal range. Infection is the most common post-OTC complication, and HSCT pretreatment can be accepted as early as the 10th day after OTC. OTC has no adverse effects on patients with hematological diseases and does not delay HSCT treatment. For young patients with hematological diseases, OTC is an effective method of fertility preservation.

Metabolic syndrome score as an indicator in a predictive nomogram for lymph node metastasis in endometrial cancer.

BACKGROUND: Lymph node metastasis (LNM) is an important factor affecting endometrial cancer (EC) prognosis. Current controversy exists as to how to accurately assess the risk of lymphatic metastasis. Metabolic syndrome has been considered a risk factor for endometrial cancer, yet its effect on LNM remains elusive. We developed a nomogram integrating metabolic syndrome indicators with other crucial variables to predict lymph node metastasis in endometrial cancer. METHODS: This study is based on patients diagnosed with EC in Peking University People's Hospital between January 2004 and December 2020. A total of 1076 patients diagnosed with EC and who underwent staging surgery were divided into training and validation cohorts according to the ratio of 2:1. Univariate and multivariate logistic regression analyses were used to determine the significant predictive factors. RESULTS: The prediction nomogram included MSR, positive peritoneal cytology, lymph vascular space invasion, endometrioid histological type, tumor size > = 2 cm, myometrial invasion > = 50%, cervical stromal invasion, and tumor grade. In the training group, the area under the curve (AUC) of the nomogram and Mayo criteria were 0.85 (95% CI: 0.81-0.90) and 0.77 (95% CI: 0.77-0.83), respectively (P < 0.01). In the validation group (N = 359), the AUC was 0.87 (95% CI: 0.82-0.93) and 0.80 (95% CI: 0.74-0.87) for the nomogram and the Mayo criteria, respectively (P = 0.01). Calibration plots revealed the satisfactory performance of the nomogram. Decision curve analysis showed a positive net benefit of this nomogram, which indicated clinical value. CONCLUSION: This model may promote risk stratification and individualized treatment, thus improving the prognosis.

Prediction of postpartum hemorrhage in pregnant women with immune thrombocytopenia: Development and validation of the MONITOR model in a nationwide multicenter study.

Globally, postpartum hemorrhage (PPH) is the leading cause of maternal death. Women with immune thrombocytopenia (ITP) are at increased risk of developing PPH. Early identification of PPH helps to prevent adverse outcomes, but is underused because clinicians do not have a tool to predict PPH for women with ITP. We therefore conducted a nationwide multicenter retrospective study to develop and validate a prediction model of PPH in patients with ITP. We included 432 pregnant women (677 pregnancies) with primary ITP from 18 academic tertiary centers in China from January 2008 to August 2018. A total of 157 (23.2%) pregnancies experienced PPH. The derivation cohort included 450 pregnancies. For the validation cohort, we included 117 pregnancies in the temporal validation cohort and 110 pregnancies in the geographical validation cohort. We assessed 25 clinical parameters as candidate predictors and used multivariable logistic regression to develop our prediction model. The final model included seven variables and was named MONITOR (maternal complication, WHO bleeding score, antepartum platelet transfusion, placental abnormalities, platelet count, previous uterine surgery, and primiparity). We established an easy-to-use risk heatmap and risk score of PPH based on the seven risk factors. We externally validated this model using both a temporal validation cohort and a geographical validation cohort. The MONITOR model had an AUC of 0.868 (95% CI 0.828-0.909) in internal validation, 0.869 (95% CI 0.802-0.937) in the temporal validation, and 0.811 (95% CI 0.713-0.908) in the geographical validation. Calibration plots demonstrated good agreement between MONITOR-predicted probability and actual observation in both internal validation and external validation. Therefore, we developed and validated a very accurate prediction model for PPH. We hope that the model will contribute to more precise clinical care, decreased adverse outcomes, and better health care resource allocation.

Curative efficacy of low frequency electrical stimulation in preventing urinary retention after cervical cancer operation.

BACKGROUND: To evaluate the clinical significance of low-frequency electrical stimulation in preventing urinary retention after radical hysterectomy. METHODS: A total of 91 women with stage IA2-IB2 cervical cancer, who were treated with radical hysterectomy and lymphadenectomy from January 2009 to December 2012, were enrolled into this study and were randomly divided into two groups: trail group (48 cases) and control group (43 cases). Traditional bladder function training and low-frequency electrical stimulation were conducted in the trail group, while patients in the control group were only treated by traditional bladder training. The general condition, rate of urinary retention, and muscle strength grades of pelvic floor muscle in the perioperative period were compared between these two groups. RESULTS: The incidence of postoperative urinary retention in the electrical stimulation group was 10.41%, significantly lower than that in the control group (44.18%), and the difference was statistically significant (P < 0.01). The duration of postoperative fever and use of antibiotics were almost the same between these two groups. Eleven days after surgery, the difference in grades of the pelvic floor muscle between these two groups was not statistically significant. However, 14 days after the operation, grades of the pelvic floor muscle were significantly higher in the trail group than in the control group, and the difference was statistically significant (P < 0.01). In addition, although there was no significant difference between the two groups with different parameters (P = 0.782), the incidence of urinary retention was lower in the endorphins analgesia program group than in the neuromuscular repair program group (9.09% < 11.54%). CONCLUSION: Low-frequency electrical stimulation is more effective than conventional intervention in preventing urinary retention after radical hysterectomy. It also intensifies the recovery of pelvic muscle strength.

Effect of transcutaneous electrical stimulation treatment on lower urinary tract symptoms after class III radical hysterectomy in cervical cancer patients: study protocol for a multicentre, randomized controlled trial.

BACKGROUND: Class III radical hysterectomy (RH III)_plus pelvic lymphadenectomy is the standard surgery for early stage cervical cancer (CC) patients, the 5 year survival rate is about 90%, but pelvic floor disorders especially bladder dysfunction are common due to damaged vessels and nerve fibers following surgery. Transcutaneous electrical stimulation (TENS) treatment has been used to treat bladder disorders for many years, but its effect on cervical cancer patients, the best treatment time point and stimulated protocol, had never been assessed. The aim of this study is to investigate the efficacy of TENS treatment on lower urinary tract symptoms (LUTS) after RH III in CC patients. METHODS/DESIGN: The study will be conducted as a clinical, multicentre, randomised controlled trial with balanced randomisation (1:1). The planned sample size is 208 participants (at 1:1 ratio, 104 subjects in each group). At 5-7 days after RH III, patients are screened according to operative and pathological findings. Enrolled participants are randomised into an intervention group (TENS plus conventional clinical care) or control group (conventional clinical care), with stratification by menopausal status (menopause vs. non-menopause) and surgical modality (laparoscopic RH or abdominal RH). Participants in both groups will be followed up at 14 days, 21 days, 28 days, 3 months, 6 months, 12 months, 18 months and 24 months after surgery. The primary endpoint is improvement rate of urination function which is defined as recovery (residual urine ≤50 ml) or improvement (residual urine 50-100 ml). Secondary endpoints include urodynamic parameter, urinary incontinence, anorectal function, pelvic function, quality of life (QOL), disease-free survival and adverse events. Primary endpoint analyses will be carried out by Cochran-Mantel-Haenszel tests taking into center effect. DISCUSSION: To our knowledge this is the first trial to investigate the effect of TENS treatment on bladder function recovery after RH III among CC patients. This study will provide new information on TENS efficacy for bladder function recovery. Once confirmed, it may help to provide a new, non-invisive treatment for those postoperative CC patients with poor pelvic function, which would help improve their quality of life. TRIAL REGISTRATION: The study is registered to Clinical Trials.gov ( NCT02492542 ) on June 25, 2015.

Ca2+ channel subunit α 1D promotes proliferation and migration of endometrial cancer cells mediated by 17ß-estradiol via the G protein-coupled estrogen receptor.

Calcium and calcium channels are closely related to the estrogen-induced nongenomic effect of endometrial carcinoma, but the specific role of calcium channels is unknown. This study aimed to explore the expression and the biologic effect of the L-type calcium channel in endometrial carcinoma cells and to clarify the molecular mechanism of the relationship between L-type calcium channels and estrogen. The immunohistochemical results showed that Ca(2+) channel subunit α 1D (Cav1.3) expression was high in atypical hyperplasia (1.90 ± 0.35) and endometrial carcinoma tissues (2.05 ± 0.82) but weak (0.80 ± 0.15) in benign endometrial tissues (P < 0.05). Treatment with 17ß-estradiol rapidly increased Cav1.3 expression in a dose- and time-dependent manner, and 100 nM cell-impermeable ß-estradiol-6-(O-carboxymethyl)oxime:bovine serum albumin also promoted Cav1.3 expression. Transfection with small interfering RNA against G protein-coupled estrogen receptor (GPER) suppressed estrogen-induced up-regulation of Cav1.3 compared with control cells and markedly reduced the estrogen-induced phosphorylation of ERK1/2 and CREB. Knocking down the Cav1.3 significantly suppressed estrogen-stimulated Ca(2+) influx, cell proliferation, and migration in endometrial cancer cells. Taken together, Cav1.3 was overexpressed in atypical hyperplasia and endometrial carcinoma, and the estrogen-induced phosphorylation of downstream molecular ERK1/2 and CREB is the result of activation of the GPER pathway. L-type channel Cav1.3 is required for estrogen-stimulated Ca(2+) influx and contributes broadly to the development of endometrial cancer. The Cav1.3 channel may be a new target for endometrial carcinoma treatment.

Outcomes of fertility and pregnancy in patients with early-stage cervical cancer after undergoing neoadjuvant chemotherapy.

OBJECTIVE: To explore the outcomes of oncology, fertility, and pregnancy in patients after undergoing neoadjuvant chemotherapy (NACT) followed by fertility-sparing operations with cervical cancer, and its value in clinical treatment. MATERIALS AND METHODS: A total of 11 patients from seven hospitals in Beijing with cervical cancer since August 2009 to December 2011, who had undergone fertility- sparing treatments were recruited in this study. RESULTS: Among the 11 patients, there were nine cases of squamous cell carcinoma, two cases of adenocarcinoma, one case in Stage IA2, and ten cases in Stage IB1 (FIGO, 2009). All of the 11 patients were treated with NACT of one to two cycles before the operations, and then they underwent radical trachelectomy (RT) + retroperitoneal lymphadenectomy. Eleven patients had completed the follow-up (100%) and the mean follow-up was 24.4 months. The outcomes of the oncology and pregnancy are as follows: no patient recurred after fertility-sparing treatments; in seven patients seeking pregnancy after the treatments, three pregnancies occurred in two women. CONCLUSIONS: NACT+RT, as a fertility-sparing treatment for young women with bulky early-stage cervical cancer and its outcomes in fertility and pregnancy are satisfactory, however its safety needs to be studied further.

[Nongenomic effects of estrogen on extracellular signal-regulated kinases through initiating transient calcium flux in endometrial cancer].

OBJECTIVE: To study the mechanism on extracellular signal-regulate kinases (ERK) signal transduction by calcium influx initiated by combination of estrogen with calcium channels or estrogen receptor in endometrial cancer cell Ishikawa. METHODS: Confocal test was used to determine the relative calcium mobilization by stimulation of estrodiol together with and without the inhibition of ICI182780 and nifedipine. Western-blotting was used to detect the protein expression of phosphorylated ERK1/2 (P-ERK1/2) in the same condition. RESULTS: The transient calcium flux initiated by 17ß-estrodiol (E2) and a membrane-impermeable conjugate of estrogen and bovine serum albumin (E2-BSA), and the calcium mobilization could be inhibited by ICI182780 and nifedipine in 1 min. In Ishikawa cells, phosphorylation of ERK1/2 was stimulated by E2, and the phosphorylation could not be inhibited by E2 after the combination with ICI182780 in 5 min and in 30 min. The phosphorylation also could not be inhibited by E2-BSA after the combination with nifedipine in 5 min, but in 30 min the phosphorylation was decreased. The phosphorylation of ERK by E2-BSA was decreased by the combination with nifedipine in 30 min. CONCLUSION: The transient calcium flux initiated by estrogen has an effect on the activation of ERK signal pathway in endometrial carcinoma cells.

[Pelvic retroperitoneal tumors: clinical analysis of 16 cases].

OBJECTIVE: To explore the diagnostic rationales for pelvic retroperitoneal tumors and summarize their therapeutic regimens. METHODS: A total of 16 retroperitoneal tumor patients were recruited. And their general information, previous medical history, physical examinations, auxiliary tests, surgical findings and postoperative pathological results were analyzed. RESULTS: Two cases were diagnosed through preoperative magnetic resonance imaging (MRI) while others found intraoperatively. Complete tumor resection was performed in all except for one case. Postoperative pathological examinations revealed 10 benign cases. And there was one case of pelvic endometriosis (mild cytologic atypia). Five cases were malignant. The operation duration was 1.45-8.5 hours and peri-operative bleeding volume 50-5000 ml. Among them, 12 patients had heavy adhesion in pelvic cavity, 7 cases underwent operations collaboratively with related departments because of surgical difficulties and vascular injury and bladder rupture occurred in 1 case. During the follow-ups, one case was lost, two patients died from disease recurrence and another one had a postoperative relapse at Month 4. The other 12 cases recovered well and had no recurrence. CONCLUSION: Surgery remains a key for retroperitoneal tumors. With a low diagnostic rate, they are often found surprisingly intraoperatively. Because of surgical difficulties and frequent complications, multi-departmental collaboration is necessary. Preoperative correct diagnosis and adequate preoperative preparation are essential. And MRI is an effective auxiliary examination.

[Analysis of relationship between patients with chromosomal translocation and the outcome of pregnancy].

OBJECTIVE: To explore the relationship between chromosome translocation and their phenotypic effect by analyzing the patients with loss pregnancy and avoiding fetuses with chromosomal abnormalities. METHODS: A total of 3067 cases with infertility or loss pregnancy were recruited to receive chromosome examination during January 2005 to December 2011 at Center of Prenatal Diagnosis, Peking University People's Hospital. Retrospective study was used to analyze the chromosome karyotypes and infertility or loss pregnancy. RESULTS: In 72 cases of patients with chromosome translocation, there were 17 pregnancies with homology translocation in fetus. And the numbers of patients with loss pregnancy and sex apparatus malformations were 40 and 15 respectively. CONCLUSION: Chromosome translocation plays an important role in patients with loss pregnancy or infertility. And chromosome examination should be performed to exclude the possibility of chromosome abnormities in patients with obstinate infertility.

[Relationship between the expression of dual specificity phosphatase-1 and the prognosis of endometrioid adenocarcinoma].

OBJECTIVE: To clarify the relationship between the expression of dual specificity phosphatase-1 (DUSP1) and the prognosis of endometrioid adenocarcinoma. METHODS: The expression of DUSP1 was determined by immunohistochemical staining in specimens from 81 patients with endometrial carcinoma undergoing surgical resection. The relationship between DUSP1 expression, clinicopathological factors and prognosis were further evaluated. RESULTS: In 81 endometrioid carcinoma samples, 59 (72.84%) cases were positive while 22 negative.Except for lymph node metastasis, the expression of DUSP1 was correlated with FIGO stage, tumor grade, myometrial invasion and the expressions of estrogen receptor (ER) and progesterone receptor (PR) (P < 0.05) .Kaplan-Meier analysis showed that patients with a positive DUSP1 expression had significantly prolonged 5-year disease-free survival rates of 98.2% and 76.0% respectively (P < 0.05). And COX regression analysis revealed that the expressions of DUSP1 and PR were independent prognostic indicators of endometrioid carcinoma, the HR (hazard ratio) of DUSP1 negative expression was 21.2.Spearman analysis further showed that the expression of DUSP1 was positively correlated with PR (r = 0.256, P < 0.05). CONCLUSION: DUSP1 may be a potential negative prognostic indicator for endometrioid adenocarcinoma.

[Preliminary study on pelvic organ prolapse treated by using small intestinal submucosa mesh].

OBJECTIVE: To study clinical efficacy of porcine-derived small intestinal submucosa (Surgisis) in treatment of pelvic organ prolapse (POP). METHODS: From Mar. 2012 to Mar. 2013, 15 patients with POP at more than III of POP quantitation (POP-Q) staging system undergoing pelvic reconstructive surgery with Surgisis in Peking University People's Hospital. The mean age was 59 years old (39-82). The variable site of POP-Q staging system was compared between preoperative and postoperative status among those patients.Quality of life questionnaire of pelvic floor impact questionnaire (PFIQ-7), pelvic floor distress inventory short form (PFDI-20) and PISQ-31 were studied to evaluate subjective satisfaction, recurrence and the quality of life's improvement. RESULTS: All patients were followed up at mean of 9.9 months (3-15 months), the mean time of surgery was 96 mins (65-120 mins), the mean blood loss was 159 ml (50-500 ml).No infection and erosion was observed on those patients. The rate of subjective satisfaction was 14/15, the recurrence rate of prolapse was 2/15.Scoring system of PFDI-20 was from 87 (56-124) at operative status to 30 (22-48) at postoperative status. PFIQ-20 was from 129 (85-158) at preoperative status to 24 (18-48) at postoperative status. PISQ-31 was from 48 (32-55) at preoperative status to 79 (66-89) at postoperative status, which all reached statistical difference (all P < 0.05). Total 9 patients obtained satisfactory sexual life. CONCLUSIONS: The short-term clinical effect of pelvic reconstructive surgery with Surgisis was satisfied, quality of life and sexual life was improved, and less complication were observed. However, long-term clinical effect on patients should be warranted to follow up.

[Study of mechanism of medroxyprogesterone 17-acetate on the cancer stem cell-like properties of human endometrial cancer].

OBJECTIVE: To explore the mechanism resistance of medroxyprogesterone 17-acetate(MPA) on the endometrial cancer side-population(SP) cells. METHODS: (1) Ishikawa-SP cells from endometrial cancer cell lines Ishikawa were be separated by Hoechst 33342 dyeing method and flow cytometry analysis. The clone formation efficiency between Ishikawa-SP cells and Ishikawa-non-SP cells were performed by clone formation assay. Breast cancer resistance protein (BCRP) was examined by immunocytochemistry method. (2) Ishikawa, Ishikawa-SP, Ishikawa-non-SP cells were treated with various concentrations of MPA at 5, 10, 15, 20 µmol/L. After cultured for 24, 48, and 72 hours, cells growth were measured by methanethiosulfomate (MTS) assay. (3) The groups of Ishikawa, Ishikawa-SP, Ishikawa-non-SP cells incubated with MPA at the half maximal inhibitory concentration (IC50) were selected for cell apoptosis assay by using flow cytometry. After MPA treatment, the expression of caspase-3 was examined by immunocytochemistry method. RESULTS: (1) There were few proportion of Ishikawa-SP cells in Ishikawa endometrial carcinoma, which were 2.7%. There were stronger clone formation efficiency for Ishikawa-SP cells than that for Ishikawa-non-SP cells in Ishikawa [(6.02 ± 1.17)% vs.(0.53 ± 0.20)%, P = 0.001]. And there were higher level expression of BCRP (P = 0.001) and also more resistant Taxol and radiation between Ishikawa-SP cells and Ishikawa-non-SP cells. (2) The inhibitory effect of MPA was concentration-dependent and time-dependent. (3)After MPA treatment, the apoptosis rates of Ishikawa-SP, Ishikawa-non-SP,Ishikawa were (4.01 ± 0.43) %, (9.30 ± 0.67) %, and (4.64 ± 0.18) %, respectively(P < 0.05). The level expression of caspase-3 in Ishikawa group after MPA treated were higher than that in Ishikawa-SP group. CONCLUSION: MPA may be inhibit the growth of endometrial cancer, Ishikawa-SP and Ishikawa-non-SP cells, while Ishikawa-SP may be more resistant to MPA than Ishikawa-non-SP, which mechanism of resistance on MPA may be related to the properties of cancer stem-like cells and cell apoptosis.

[Application of endometrial sampling device during the follow-up visit for the conservative treatment of endometrial cancer].

OBJECTIVE: To study the feasibility of endometrial sampling device as a sampling tool during the follow-up visit for endometrial cancer patients undergone conservative treatment. METHODS: Before the hysteroscopy examination, endometrial sampling device was used to take the endometrium specimens 43 times in 19 patients who had been diagnosed as endometrial cancer or atypical hyperplasia, and were undergone conservative treatment during May 2012 to Mar. 2013. All cases accepted vaginal ultrasound screening before every sampling by endometrial sampling device. The histological results were compared with those done by hysteroscopy. RESULTS: The average age of those patients was (30 ± 6) years old. The mean thickness of the endometrium during the treatment was (0.81 ± 0.65) cm. The qualified rate for the sampling was 95% (41/43). Compared with the specimens undergone by hysteroscopy direct sampling, 32 samples got by the endometrial sampling device with thicker endometrium (0.93 ± 0.70) cm had the same histological results, while the other 9 patients with thinner endometrium (0.40 ± 0.14) cm were not (P = 0.031). CONCLUSION: The endometrial sampling device could be used during the follow-up visit for the conservative treatment patients with endometrial cancer or atypical hyperplasia, the vaginal ultrasound screening should be used together to figure out those with thinner endometrium.

[Efficacy of Le Fort operation and its impact on patients' quality of life].

OBJECTIVE: To evaluate the objective and subjective effects of Le Fort operation and its impact on patients' quality of life before and after operation. METHODS: A total of 48 patients with pelvic organ prolapse undergoing Le Fort operation from October 2005 to May 2010 were recruited. Their clinical data were retrospectively analyzed and regular follow-ups of pelvic examinations and questionnaires conducted. The questionnaires included pelvic floor distress inventory-short form 20 (PFDI-20) and pelvic floor distress impact questionnaire-short form 7 (PFIQ-7). RESULTS: Forty patients had complete follow-up data. Their mean follow-up period was 22.3 months (range: 12 - 42). Among them, there were recurrence (n = 1), new-onset stress urinary incontinence symptoms (n = 6) and new-onset colorectal-anal symptoms (n = 2). The postoperative scores of PFDI-20, PFIQ-7, UDI-6 and POPDI-6 decreased markedly (P < 0.05). CONCLUSION: Le Fort operation can significantly improve patients' quality of life and subjective prolapse symptoms and lower urinary tract. However its improvement of colorectal-anal symptoms remains to be defined.

[Survey of long term female pelvic floor function and sexual life status after total hysterectomy].

OBJECTIVE: To evaluate status of female pelvic floor function and sexual life after total hysterectomy. METHODS: From March 2001 to January 2004, 92 patients with undergoing hysterectomy due to benign gynecological diseases were enrolled in this study. They were followed up at outpatient department, including pelvic examination, filling in female sexual quality questionnaire, pelvic floor distress inventory short form 20 (PFDI-20), pelvic floor distress impact questionnaire short form 7 (PFIQ-7) and quality of sexual life of chinese women questionnaire. RESULTS: At 6 years after total hysterectomy, it was observed that 7 cases (7/92, 7.6%) were pelvic organ prolapse and 62 cases (62/92, 67.4%) were urinary incontinence. A median score of PFDI-20 were 4.67. A median score of PFIQ-7 were 0. Symptoms of pelvic floor dysfunction concentrated in lower urinary tract (58 cases with cough leak, 32 cases with spot urine leakage, 31 cases with frequent micturition, 24 cases with urgent urination)and bowel symptoms (26 cases with constipation, 24 cases with defecation urgency, 21 cases without fully drained stool). In the 68 patients filling in female sexual quality questionnaire, an average score were (127 ± 20) points. Female sexual quality questionnaire score, sexual satisfaction, sexual communication and adjustment, sexual response and sexual body image were positively correlated with the patients' income (r = 0.432, P = 0.007; r = 0.356, P = 0.028; r = 0.475, P = 0.003; r = 0.421, P = 0.009; r = 0.324, P = 0.047). CONCLUSIONS: Hysterectomy may have long-term effect on female pelvic floor function and sexual life. Quality of sexual life in those patients was positively correlated with income.

[Study of the invasiveness and tumour formation of Bcrp1(+) HeLa cervical cancer cells].

OBJECTIVE: To make sure whether or not Bcrp1 is the marker of cervical cancer stem cells or not by studying the invasive ability and formation of tumors of Bcrp1(+) phenotype HeLa cells. METHODS: The tumor cell migration and invasion assay were used by boyden chamber to identify the invasive ability of Bcrp1(+) phenotype HeLa cells. The formation of tumors in vivo experiments were completed, in which the two groups of cells with different concentrations were inoculated in non obese diabetes-severe combined immunodeficiency disease (NOD/SCID) mice (1×10(4), 1×10(5), 1×10(6)/ml), and the differences of time, rate and volume in the formation of tumors between two groups were observed. RESULTS: (1) In the invasion assay, the amount of cells that invaded through the artificial basement membrane in Bcrp1(+) group were 99 ± 14, which was significantly greater than those in Bcrp1(-) group (57 ± 13, P < 0.05);the length of the Bcrp1(+) group was (366 ± 52) µm, which was significantly greater than the Bcrp1(-) group (301 ± 54) µm (P < 0.05). (2) Following transplantation of 1×10(4) cells, only the Bcrp1(+) cells formed tumors in NOD/SCID mice. When 1×10(5) or 1×10(6) cells were transplanted, the tumor incidence and the tumor mass were greater in the Bcrp1(+) groups than those in the Bcrp1(-) groups (P < 0.05). CONCLUSION: Bcrp1(+) HeLa cell have the greater capacity of invasive and the tumorigenicity, which may contain cancer stem cells.

[Impact of calcium channel antagonists for estrogen action on the endometrial carcinoma HEC-1A cells].

OBJECTIVE: To study the effects of nifedipine and mibefradil on the cell proliferation, apoptosis, migration on the HEC-1A in vitro and also study the mRNA and protein expression levels of the calcium channel alpha1D (Cav1.3) and calcium channel alpha1G (Cav3.1) to discuss the effects of the calcium antagonists on the mechanisms of the endometrial carcinoma. METHODS: (1) Add 10 µmol/L nifedipine and mibefradil at 15 minutes before adding 10 µmol/L 17ß-estradiol (17ß-E(2)) and 100 µmol/L b-estradiol-6-(O-carboxymethyl)oxime (E(2)-BSA) to the HEC-1A in different time including 0, 5, 15, 30, 60, 120 minutes. Then the changes of mRNA and protein were detected by reverse transcripiton (RT)-PCR and western-blot. (2) Add 1.25, 2.5, 5, 10, 20, 40, 80, 100 µmol/L nifedipine and mibefradil to the HEC-1A at 24, 48, 96 hours to detect the cell proliferation by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) method. (3) Add 10 µmol/L nifedipine and mibefradil to the HEC-1A, then detect the apoptosis at 0 minute, 30 minutes, 1 hour, 6 hours, 24 hours and migration in vitro at 36 hours with transwell methods. RESULTS: (1) After the pretreated effect of the nifedipine before 17ß-E(2), the mRNA express of Cav1.3 genes was lowest at 15 minutes, and returned to the control level after 30 minutes. The protein level didn't change very much in 30 minutes, but rose after 60 minutes. The Cav3.1 genes mRNA express was lowest at 5 minutes, rose at 30 minutes and returned to the 0 minute level gradually. (2) After the pretreated effect of the nifedipine before E(2)-BSA, the Cav1.3 genes mRNA was lowest at 5 minutes and returned at 15 minutes. The protein level rose gradually in 15 minutes but reduced after 15 minutes. The Cav3.1 mRNA and protein level were reduced at every time point. (3) After the pretreated effect of the mibefradil before 17ß-E(2), there was no change of mRNA expression of Cav1.3 genes. The protein level rose at 15 and 60 minutes, there was no change in any other time. The Cav3.1 genes mRNA were gradually reduced and the protein level rose at 15 minutes, and there was no change in any other times. (4) After the pretreated effect of the mibefradil before E(2)-BSA, the mRNA and protein of Cav1.3 levels were reduced after 15 minutes. There was no mRNA expression of Cav3.1, while the protein level was lowest at 15 minutes. (5) Nifedipine and mibefradil affected HEC-1A proliferation depended on the different concentration and interval time points. There was significant difference than those in control group (P < 0.05). (6) There were statistical differences in apoptosis rate after adding nifedipine (P < 0.05), while rose at mibefradil treated the same time (24 hours: 8.41 ± 0.07, 0 minute: 3.74 ± 0.18; P < 0.05). (7) The numbers of stained cells after both nifedipine and mibefradil treated reduced more than control group. CONCLUSIONS: (1) Nifedipine and mibefradil could inhibit both the effect of the estrogen on the L-type and T-type calcium channel in short time, meanwhile the mibefradil effects last long time. (2) The inhibited effect of the mibefradil on the proliferation, apoptosis and migration of HEC-1A cells in vitro is more significant than that by nifedidipine.

[Preliminary investigation of the expression and functions of insulin receptor isoforms in endometrial carcinoma].

OBJECTIVE: To investigate the expression of insulin receptor isoforms (IR, including IR-A and IR-B) in endometrial carcinoma (EC), and explore the role of IR-A in the growth of endometrial carcinoma cells. METHODS: The expression of IR isoforms were detected by reverse transcription (RT)-PCR and real-time PCR in 4 different endometrial cancer cell lines (HEC-1-A, Ishikawa, RL95-2 and KLE), with human breast cancer cell line MCF-7 cells and hepatocellular carcinoma cell line Hep-G2 as positive control and in the endometrial cancer tissue specimens of 42 cases, admitted in Peking University People's Hospital from November 2007 to July 2009, with normal endometrial tissues from 15 cases of ovarian neoplasms at the same period as controls. The relationships among the expression of IR, IR-A isoforms and the clinicopathological parameters of EC tissues were analyzed by Spearman rank correlation analysis. An eukaryotic IR-A expression plasmid was constructed and transfected into RL95-2 cells [RL95-2(IR-A)] for overexpression of IR-A in RL95-2 cells, in which the expression of IR-A originally was low. Non radioactive cell proliferation assay-MTS was used to determine the proliferation curves in the four EC cells listed above and in RL95-2 or RL95-2 (IR-A). RESULTS: (1) There were two isoforms of IR-A and IR-B co-expressed were detected in EC cells and EC tissues. Among four kinds of EC cell lines, the expression level of IR mRNA in RL95-2 cells was the highest, followed by Ishikawa, KLE and HEC-1-A cells, in which the relative IR mRNA expression levels were (26.54 ± 1.82)×10(-4), (15.44 ± 3.29)×10(-4), (10.14 ± 0.10)×10(-4) and (2.63 ± 0.23)×10(-4), respectively (P < 0.01). The expression level of IR-A mRNA was the highest in Ishikawa cells, followed by KLE, RL95-2 and HEC-1-A cells, with the relative expression levels were (12.07 ± 3.31)×10(-4), (4.68 ± 0.63)×10(-4), (3.03 ± 0.22)×10(-4) and(1.46 ± 0.03)×10(-4), respectively (P < 0.01). The relative expression level of IR and IR-A mRNA were 0.017 ± 0.013 and 0.011 ± 0.010 in the EC tissues, respectively, compared with 0.015 ± 0.014 and 0.010 ± 0.012 in the controls, in which there were no significant differences in the expression level of IR or IR-A mRNA between EC tissues and the control (P = 0.662, P = 0.780). The expression of IR and IR-A in EC tissues had no significant relevance with International Federation of Gynecology and Obstetrics (FIGO) stage, cell differentiation, depth of myometrial invasion, invasion of lymph-vascular space, lymph nodes metastasis, the expression status of estrogen receptor, human progesterone receptor, and PTEN gene (all P > 0.05). The expression of IR-A mRNA in EC patients with type 2 diabetes mellitus (DM) was significantly higher than that in patients without type 2 DM (P = 0.031), while there were no statistical correlation between the expression of IR mRNA and type 2 DM (P = 0.438). (2) The proliferation rates of the four kinds of EC cells was positively related with the IR-A expression ratio, with the most growth potential in Ishikawa, followed by HEC-1-A, KLE and RL95-2 cells. The overexpression of IR-A in RL95-2 (IR-A) cells showed a significant proliferation-promoting effect than that in control RL95-2 cells (P < 0.01). CONCLUSION: There are two isoforms of IR-A and IR-B co-expressed in EC. The overexpression of IR-A may promote the proliferation of EC cells.

[Analysis of the perinatal outcome and risk factors for pregnancies complicated with chronic renal diseases].

OBJECTIVE: To investigate the perinatal outcome for pregnancies complicated with chronic renal diseases, and the risk factors for the adverse outcome. METHODS: Retrospectively analyze the clinical data of 48 patients with chronic renal diseases complicating pregnancy admitted in Peking University People's Hospital between January 1998 and August 2010, record the pregnancy outcome and explore the risk factors for the poor outcome using multivariate regression analysis. RESULTS: Thirty-eight patients had known chronic renal disease before conception, and ten were diagnosed during pregnancy. Seven patients (15%, 7/48) presented with obvious renal impairment [serum creatinine (sCr) ≥ 125 µmol/L] prepregnancy, and nine (19%, 9/48) were recorded with chronic hypertension. Thirty-three patients received regular prenatal care. Twenty-one cases (44%, 21/48) developed preeclampsia. During the gestation, normal renal function (defined as sCr < 71 µmol/L) was seen in nineteen cases (40%, 19/48), mild dysfunction (sCr ranged 71 - 132 µmol/L) in twenty (42%, 20/48) and moderate to severe dysfunction (sCr ≥ 132 µmol/L) in nine cases (19%, 9/48). Twenty patients had negative or mild proteinuria (24 hour urine protein < 2000 mg), 19 had moderate (24 hour urine protein ranged 2000 - 5000 mg) and nine had severe proteinuria (24 hour urine protein ≥ 5000 mg). The gestational age at delivery ranged from 24 to 41 weeks and the neonatal birth weight ranged from 890 to 4150 g. A total of twenty patients (42%, 20/48) suffered adverse perinatal outcome, including one case with late spontaneous abortion, fifteen with preterm delivery, eleven with small for gestational age, two with neonatal respiratory distress syndrome and four with perinatal death. Declined maternal renal function was seen in eight patients, and two patients progressed toward the end-stage renal failure (the stage of uremia). Multivariate regression analysis identified that preeclampsia (OR = 24.72, P = 0.002) and the degree of proteinuria (OR = 4.24, P = 0.032) were the independent risk factors for the adverse perinatal outcome. CONCLUSIONS: Pregnancies complicated with chronic renal diseases have significantly high incidence of preeclampsia and adverse perinatal outcome. Preeclampsia and the degree of proteinuria are perhaps the independent risk factors for the adverse outcome.