Hyperlipidemia is negatively associated with pregnancy outcomes in patients following frozen-thawed embryo transfer: A retrospective study.

Background and Aims: It is demonstrated that lipid metabolism disorders are associated with the reproductive performances of assisted reproductive technology. However, it is little known whether hyperlipidemia is associated with the endometrial receptivity and pregnancy outcomes of patients undergoing frozen-thawed embryo transfer (FET). Methods: This was a retrospective analysis involving 554 infertile women undergoing FET. The patients were divided into the hyperlipidemia group (n = 224) and control group (n = 320) based on the levels of serum lipids. The clinical and laboratory indexes between the two groups were compared. Meanwhile, the stratified analysis based on body mass index (BMI) and endometrial preparation protocols was performed. The independent samples t-test, Mann-Whitney U test, χ2 test and multiple logistic regression analysis were used to compare and analyze the data. Results: The patients with hyperlipidemia had significantly higher serum lipids levels and BMI and lower clinical pregnancy and implantation rates than those with normal blood lipids (p < 0.05). The impact of hyperlipidemia on pregnancy outcomes was independent of BMI. The multiple logistic regression analysis showed that higher cholesterol was associated with lower pregnancy rate and implantation rate (p < 0.05). Regardless of blood lipid levels, the patients undergoing the hormone replacement therapy (HRT) protocol had higher estradiol levels and lower progesterone levels compared with the stimulated cycles (STC) (p < 0.05). Moreover, the clinical pregnancy rate and implantation rate of the HRT protocol were higher than those of the STC, although there was no significant difference between the two. Conclusion: Hyperlipidemia especially higher cholesterol has a negative effect on the pregnancy outcomes of the patients undergoing FET. Actively implementing lipid-lowering treatment and the HRT protocol seem more friendly for these patients.

Effect of hyperlipidemia on the outcome of in vitro fertilization in non-obese patients with polycystic ovary syndrome.

Introduction: It is little known whether hyperlipidemia alone has adverse effects on the outcome of in vitro fertilization (IVF) in patients with polycystic ovarian syndrome (PCOS). Methods: The PCOS patients with body mass index (BMI) < 30 kg/m2 were performed IVF or intracytoplasmic sperm injection treatment, including 208 fresh cycles and 127 frozen embryo transfer (FET) cycles. All the patients were divided into hyperlipidemia and control groups, and embryo quality and pregnancy outcomes between the two groups were compared. Results: In the fresh cycles, total gonadotropin dosage in the control group was significantly lower than that in the hyperlipidemia group, and serum estradiol levels on trigger day were reversed (P < 0.05). The embryo fragment score was positively correlated with serum low-density lipoprotein level (r = 0.06, P < 0.05) and negatively with serum high-density lipoprotein (HDL) and lipoprotein A levels (r = -0.489 and -0.085, P < 0.01). Logistic regression analysis found that HDL was beneficial for clinical pregnancy (OR = 0.355, 95% CI: 0.135-0.938, P < 0.05). In the FET cycles, there were no differences in pulse index, systolic/diastolic ratio and serum estradiol and progesterone levels between the two groups, but resistance index in the hyperlipidemia group was significantly higher than that in the control group (P < 0.05). Conclusion: Hyperlipidemia may increase the dosage of gonadotropin and have adverse effect on the embryo quality, endometrial receptivity, and clinical outcomes of lean PCOS patients. It is recommended that the non-obese patients with hyperlipidemia and PCOS perform lipid-lowering treatment before undergoing embryo transfer.

Performance evaluation of sperm concentration, motility, and morphological analysis for GSA-810 series of sperm quality analysis system.

BACKGROUND: The performance evaluation of each computer-assisted sperm analysis (CASA) system may provide a basis for the interpretation of clinical results and further improvement of the CASA system. METHODS: The accuracy of the GSA-810 CASA system was evaluated by detecting latex bead quality control products. The precision of sperm concentration, morphology, and percentages of progressively motile sperm (PR) were evaluated by coefficient of variation (CV). Three samples with sperm concentration of about 100 × 106 /mL were diluted to evaluate the linear range. RESULTS: The detection values of latex beads were within the range of target values. The CVs of sperm concentration and PR were significantly and negatively correlated with sperm concentration (r = -0.561, p = 0.001) and PR value (r = -0.621, p < 0.001), respectively. The R2 values of the linear range of sperm concentration were ≥0.99. There was no significant difference in sperm motility and PR within 1-10 min at 36.5°C ± 0.5°C. The coincidence rates of sperm morphology and sperm head morphology for 36 semen samples analyzed by the GSA-810 system and manual method were 99.40% and 99.67%, respectively. The CVs of the percentage of sperm with abnormal morphology and percentage of sperm with abnormal head morphology were less than 5%. CONCLUSION: The GSA-810 system can accurately analyze normal semen samples, but the repeatability of the results is poor for oligozoospermia and asthenozoospermia samples. The future CASA system for analyzing sperm morphology should focus on recognizing the middle and tail segments of a spermatozoon.

Analysis of selected sperm samples by a computer-assisted system with high frame rate: A prospective study.

Background and Aims: Due to the rapid motility of the selected sperm, sperm parameters cannot be accurately determined by the manual method. So, the application of a computer-assisted sperm analysis system with a high frame rate (HFR-CASA, 85 Hz) in sperm selection is investigated. Methods: A total of 177 semen samples were collected for sperm selection with density gradient centrifugation. Then, the manual method and HFR-CASA method will be used to observe and analyze the sperm concentration, motility, and percentage of progressively motile sperm (PR) of the selected sperm samples. The quality control of sperm concentration was performed with microballoons. Two selected sperm samples were analyzed 10 times repeatedly to evaluate the accuracy of the HFR-CASA. Results: The results of microballoons analyzed with the HFR-CASA were in control. The coefficients of variation of sperm concentration, motility, and PR from two selected sperm samples were all below 10%. The results of 177 selected sperm samples analyzed by the manual method and HFR-CASA showed that although there were significant positive correlations in sperm concentration, motility, and PR between them (p < 0.001), the manual method significantly underestimated sperm concentration (p = 0.002) but overestimated sperm motility and PR (p < 0.001). When sperm concentration was below 50 × 106/mL, the manual method significantly overestimated sperm concentration (p < 0.05). However, when sperm concentration was more than 100 × 106/mL, the manual method significantly underestimated sperm concentration (p < 0.001). Regardless of sperm concentration, the manual method consistently overestimated sperm motility and PR (p < 0.001). When sperm concentration was more than 20 × 106/mL, there was no correlation in sperm PR between them (p > 0.05). When sperm concentration was below 50 × 106/mL, the correct rate of captured sperm by the HFR-CASA was more than 98%. Conclusion: The HFR-CASA method is more accurate than the manual method in analyzing the selected sperm samples.

Correlations of joint detection of 22 vaginal microbes with routine examination results of vaginal secretions and assisted reproductive outcomes.

The correlations of joint detection of 22 vaginal microbes with routine examination results of vaginal secretions and assisted reproductive outcomes were investigated. There were 37 samples with abnormal vaginal microecology in 107 vaginal secretion samples. The top 5 detection rates of microorganisms were Ureaplasma urealyticum (73.83%), Prevotella sp. (70.09%), Gardnerella vaginalis (53.27%), L. crispatus (52.34%) and L. inerts (51.40%). When the levels of Bacillus and hydrogen peroxide in vaginal secretions decreased or pH increased, the abnormal rates of vaginal microecology increased significantly (P < 0.01). The clinical pregnancy rate (53.66%, 22/41) in the women with normal vaginal microecology was higher than that (37.5%, 9/24) with abnormal vaginal microecology. In conclusions, the joint detection of 22 vaginal microbes can quickly and effectively determine whether the vaginal microecology is normal or not. The evaluation of vaginal microecology may be valuable in predicting the assisted reproductive outcomes of infertile women.

Screening of sperm antigen epitopes by phage display technique and its preliminary clinical application.

BACKGROUND: At present, there is a lack of standardized preparation methods of sperm antigen for the detection of antisperm antibody (AsAb). To screen sperm antigen mimotopes from a phage display random peptide library and use them to establish an enzyme-linked immunosorbent assay (ELISA) for the detection of AsAb, immunoglobulins were extracted from the sera of rabbits with positive AsAb and negative AsAb, respectively, by the saturated ammonium sulfate method, and a phage display 12-mer peptide library was affinity panned by the extracted immunoglobins coated on the ELISA plate. Then, the obtained positive phage clones were identified by ELISA and sent for sequencing and peptides synthesis. Last, a diagnostic ELISA was established to detect clinical serum and seminal plasma samples. RESULTS: A total of sixty phage clones were chosen by affinity panning, and sixteen of them reacted positively with AsAb in indirect ELISA and sandwich ELISA. Following DNA sequencing and translation, the peptide sequences of the sixteen positive clones were obtained. By comparison in Blast database, four of sixteen positive clones were found to be closely related to male reproduction. Two (#1 and #25) of four mimotopes were synthesized, and an ELISA method was established using the two mimotopes as sperm specific antigens. One hundred and thirty-four serum samples and seventy-four seminal plasma samples from infertile couples were analyzed by the established ELISA with #1 and #25 mimotopes, respectively. The positive rates of AsAb in serum samples were 20.15% (27/134) for #1 and 11.19% (15/134) for #25, respectively, and the coincidence rate between them was 91.04% (122/134). The positive rates of AsAb in seminal plasma samples were 1.35% (1/74) for both #1 and #25, and the coincidence rate was 100%. CONCLUSION: Sperm antigen mimotopes can be obtained successfully by the phage display technique, and can be used as standard sperm specific antigens to establish an ELISA method for the detection of AsAb.

RéSUMé: CONTEXTE: À ce jour, il n'existe pas de méthodes normalisées de préparation d'antigènes spermatiques pour la détection des anticorps anti-spermatozoïdes (ACAS). Dans le but d'élaborer un tel test ELISA (enzyme-linked immunosorbent assay), nous avons extrait de sérum de lapins des anticorps anti-spermatozoïdes humains via la technique du sulfate d'ammonium saturé et en ayant recours à une librairie phagique de peptides (12-mer). Les clones positifs ont été identifiés par ELISA, séquencés à façon et les peptides correspondants ont été synthétisés. In fine, un test ELISA diagnostic a été conçu pour être utilisé avec des échantillons cliniques de sérum et de plasmas séminaux. RéSULTATS: Au total, soixante clones de phages ont été sélectionnés, et seize d'entre eux se sont avérés interagir avec les ACAS en ELISA indirect comme en ELISA sandwich. Les séquences peptidiques de ces seize clones positifs ont été obtenues. Par comparaison avec les bases de données (Blast), quatre de ces seize clones positifs se sont révélés être étroitement liés à la reproduction masculine. Deux des quatre mimotopes (#1 et #25) ont été synthétisés, et un test ELISA a été généré en utilisant ces deux mimotopes comme antigènes spécifiques des spermatozoïdes. Cent trente-quatre échantillons de sérum et soixante-quatorze échantillons de plasma séminal de patients de couples infertiles ont alors été analysés avec ce test ELISA. Respectivement, les échantillons sériques se sont révélés positifs à 20,15% (27/134) pour le mimotope #1 et à 11,19% (15/134) pour le mimotope #25, avec un taux de coïncidence de 91,04% (122/134). Seul un échantillon de plasma séminal (1/74, soit 1, 35%) s'est révélé positif à la fois pour le mimotope #1 et #25 (coïncidence 100%). CONCLUSION: La technique « phage display¼ nous a permis d'identifier des mimotopes d'antigènes spermatiques qui ont pu être utilisés afin de générer un test ELISA pour la détection d'anticorps anti-spermatozoïdes.

Analysis of human sperm DNA fragmentation index (DFI) related factors: a report of 1010 subfertile men in China.

BACKGROUND: Many factors may lead to sperm DNA damage. However, it is little known that the correlations of sperm DNA damage with obesity-associated markers, and reproductive hormones and lipids levels in serum and seminal plasma. METHODS: In our prospective study, a total of 1 010 subfertile men, aged from 18 to 50 years old, were enrolled from August 2012 through June 2015. Their obesity-associated markers, semen parameters, sperm acrosomal enzyme activity, seminal plasma biochemical markers, and reproductive hormones and lipids levels in serum and seminal plasma were detected. Sperm DNA fragmentation index (DFI) was determined by sperm chromatin structure assay. The correlations between DFI and each of the above-mentioned variables were analyzed. RESULTS: Spearman correlation analysis showed that sperm DFI was positively related to age and abstinence time (P<0.001). Sperm DFI was also positively related to semen volume and percent of abnormal sperm head (P<0.001), while negatively related to sperm concentration, progressive motility (PR), sperm motility, total normal-progressively motile sperm count (TNPMS), percent of normal sperm morphology (NSM), percent of intact acrosome and acrosomal enzyme activity (P<0.001). Sperm DFI was positively related to seminal plasma zinc level (P<0.001) but unrelated to seminal plasma total α-glucotase, γ-glutamyl transpeptidase (GGT) and fructose levels. There was no any correlation between sperm DFI and obesity-associated markers such as body mass index (BMI), waist-to-hip ratio (WHR), waist circumference (WC) and waist-to-height ratio (WHtR), and serum lipids levels, but there was positive correlation between sperm DFI and seminal plasma triglyceride (TG) and total cholesterol (TC) levels (P<0.001). Sperm DFI was positively related to serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels and seminal plasma FSH and estradiol (E2) levels (P<0.001), but unrelated to serum and seminal plasma testosterone (T) levels. The multivariate regression analysis for the variables which were significantly correlated with sperm DFI in Spearman correlation analysis showed that age, semen volume, sperm concentration, progressive motility, TNPMS and intact acrosome were independently correlated with sperm DFI. CONCLUSIONS: There are many potential factors associated with sperm DFI, including age, abstinence time, spermatogenesis and maturation, seminal plasma lipids and reproductive hormones levels. However, the potential effects of seminal plasma lipids and reproductive hormones on sperm DNA damage need still to be demonstrated by the studies with scientific design and a large size of samples.

[Value of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility].

OBJECTIVE: To investigate the clinical significance of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility (UI). METHODS: This retrospective study included 49 UI couples treated by IVFET (49 cycles) after three failures in intrauterine insemination (IUI) and another 95 couples with uterine tube obstruction (UTO) treated by IVF (131 cycles). We analyzed the laboratory data, clinical outcomes and sperm acrosin activity in the two groups of patients. According to the level of sperm acrosin activity of the males, we further divided the UI patients into two subgroups, a < 36 IU/106 sperm group (20 cycles) and a ≥36 IU/106 sperm group (29 cycles), and compared the fertilization rates between the two groups. RESULTS: Compared with UI couples treated by IVFET, the UTO couples treated by IVF had a significantly lower rate of fertilization (67.0% vs 76.4%, P < 0.05) and a higher rate of remedial intracytoplasmic sperm injection (ICSI) (20.4% vs 6.1%, P < 0.05), but showed no statistically significant differences in the rates of MII oocytes, available embryos, highquality embryos, implantation, and clinical pregnancy from the latter group (P >0.05). The sperm acrosin activity was remarkably lower in the UI than in the UTO patients (36.03 vs 61.98 IU/106, P < 0.01), and so was the fertilization rate in the < 36 IU/106 than in the ≥36 IU/106 sperm subgroup (47.7% vs 80.3%, P < 0.01). CONCLUSIONS: The low fertilization rate caused by decreased sperm acrosin activity may be the main cause of infertility and the potential factor of UI. When sperm acrosin activity is < 36 IU/106 sperm, IVF plus shortterm fertilization by remedial ICSI should be preferred to IUI.

Electroacupuncture decreases the progression of ovarian hyperstimulation syndrome in a rat model.

This study aimed to elucidate the effect of electroacupuncture treatment on preventing early ovarian hyperstimulation syndrome (OHSS) and the potential mechanisms involved using an induced rat model. The ovarian response was examined by measuring ovary weight, vascular permeability, levels of inflammation (interleukin-6), tumour necrosis factor alpha, chemokine ligand 2 (also known as monocyte chemoactic protein 1), vascular endothelial growth factor and hormone concentrations (oestradiol, progesterone, testosterone and prolactin). Sprague-Dawley female rats underwent ovarian stimulation to induce OHSS. Hyperstimulated rats received consecutive electroacupuncture treatment from 3 days before the beginning of pregnant mare serum gonadotrophin treatment or the time point of pregnant mare serum gonadotrophin treatment respectively, and last until 3 days after HCG administration. Electroacupuncture treatment reduced ovary weight and vascular permeability in hyperstimulated rats. Electroacupuncture treatment also reduced the levels of serum steroid hormones (progesterone and testosterone), inflammatory cytokines (interleukin-6, tumour necrosis factor alpha and monocyte chemotactic protein 1 and vascular endothelial growth factor in hyperstimulated rats. The results indicate that electroacupuncture can modulate endocrine hormone secretion and affect the secretion of inflammatory cytokines and vascular endothelial growth factor, and thus prevent the progress of OHSS. Electroacupuncture may provide a simple and effective method for the prevention and treatment of OHSS.

Relationship between Lipids Levels of Serum and Seminal Plasma and Semen Parameters in 631 Chinese Subfertile Men.

OBJECTIVE: This prospective study was designed to investigate the relationship between lipids levels in both serum and seminal plasma and semen parameters. METHODS: 631 subfertile men were enrolled. Their obesity-associated markers were measured, and semen parameters were analyzed. Also, seminal plasma and serum TC, TG, HDL and LDL and serum FFA, FSH, LH, total testosterone (TT), estradiol (E2) and SHBG levels were detected. RESULTS: Seminal plasma and serum TG, TC and LDL levels were positively related to age. Serum TC, TG and LDL were positively related to obesity-associated markers (P < 0.001), while only seminal plasma TG was positively related to them (P < 0.05). For lipids levels in serum and seminal plasma, only TG level had slightly positive correlation between them (r = 0.081, P = 0.042). There was no significant correlation between serum lipids levels and semen parameters. However, seminal plasma TG, TC, LDL and HDL levels were negatively related to one or several semen parameters, including semen volume (SV), sperm concentration (SC), total sperm count (TSC), sperm motility, progressive motility (PR) and total normal-progressively motile sperm counts (TNPMS). Moreover, seminal plasma TG, TC, LDL and HDL levels in patients with oligospermatism, asthenospermia and teratozoospermia were higher than those with normal sperm concentration, motility or morphology. After adjusting age and serum LH, FSH, TT, E2 and SHBG levels, linear regression analysis showed that SV was still significantly correlated with seminal plasma LDL (P = 0.012), both of SC and TSC with seminal plasma HDL (P = 0.028 and 0.002), and both of PR and sperm motility with seminal plasma TC (P = 0.012 and 0.051). CONCLUSION: The abnormal metabolism of lipids in male reproductive system may contribute to male factor infertility.

Annexin A5 regulates Leydig cell testosterone production via ERK1/2 pathway.

This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig cells were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory (StAR), P450scc, 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), and 17α-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of StAR, P450scc, 3ß-HSD, and 17ß-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3ß-HSD, and 17ß-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 (50 µmol l-1 ) for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3ß-HSD, and 17ß-HSD were completely abrogated (P < 0.05). Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3ß-HSD, and 17ß-HSD in Leydig cells.

[Melatonin regulates ovarian function: an update].

Melatonin (N-acetyl-5-methoxytryptamine, MT) is a hormone synthesized and secreted by the pineal gland. Recent studies show that melatonin plays an essential role in the pathogenesis of many reproductive processes. High-concentration melatonin exists in human preovulatory follicular fluid and melatonin receptors are present in ovarian granulosa cells, which indicates the direct effects of melatonin on ovarian function. Reactive oxygen species are involved in a number of reproductive events, including folliculogenesis, follicular atresia, ovulation, oocyte maturation, and corpus luteum formation. Melatonin and its metabolites, as powerful antioxidants and free radical scavengers, can potentially inhibit premature ovarian failure. Literature published in recent years shows the essential roles of melatonin in improving human ovarian function and oocyte quality as well as in the management of infertility. Researches on the action mechanisms of melatonin may provide a theoretical basis for the prevention and treatment of some clinical diseases.

[Effect of StarD7 and Wnt/ß-catenin signal pathway on the testosterone secretion stimulated by Annexin 5 in rat Leydig cells].

OBJECTIVE: This research aims to study the internal mechanism that promotes the testosterone synthesis by StarD7 and Wnt/ß-catenin, and explores a new regulatory pathway of testosterone synthesis. METHODS: After treated with 1 nmol/L Annexin 5 for 24 h, the culture media were collected for testosterone measurement by chemiluminescence assay. The expressions of StarD7 and ß-catenin at mRNA and protein levels were detected by RT-PCR and western blot respectively. The cellular location of ß-catenin was identified by immunofluorescence. RESULTS: Comparing with the control groups, under the treatment with Annexin 5, the level of testosterone raised 176%[(7.83±0.32)vs.(21.6±1.1), P<0.05], StarD7 mRNA in the experimental groups increased 55%[(1.12±0.08)vs.(1.74±0.11), P<0.05], and ß-catenin mRNA increased 48%[(1.15±0.08)vs.(1.70±0.05), P<0.05]. At the level of protein, the expression of StarD7 in the experimental groups increased 42%[(1.06±0.09)vs.(1.51±0.07), P<0.05], and ß-catenin increased 55%[(1.02± 0.01)vs.(1.58±0.02), P<0.05]. Immunofluorescence identified that ß-catenin was accumulation in the nuclear of the rat Leydig cells in the experiment groups cultured with Annexin 5. CONCLUSION: StarD7 and ß-catenin have both increased significantly at the mRNA and protein levels under treatment with the Annexin 5, and ß-catenin were accumulation in the nuclear of the rat Leydig cells. It suggests that StarD7 and ß-catenin both regulate the effect of Annexin 5 in testosterone production of rat Leydig cells. This regulation may active the Wnt/ß-catenin signal pathway, then increase the expression of the StarD7, eventually raise the progress of the testosterone secretion in rat Leydig cells.

Insulin-like growth factors in endometrioid adenocarcinoma: correlation with clinico-pathological features and estrogen receptor expression.

BACKGROUND: Endometrial carcinoma is a common malignancy of female genital tract. Insulin-like growth factor is known to elicit estrogen-induced mitogenic activity and anti-apoptotic effect in endometrial tissues. METHODS: The retrospective study investigated the expression of insulin-like growth factors, estrogen receptors and their associations in endometrioid adenocarcinoma (EAC) from 80 EAC patients in immunohistochemistry, and 58 EAC patients and 42 control patients in quantitative RT-PCR. The Pearson correlation analysis was used to analyze their correlations with clinic-pathological parameters. RESULTS: Our results showed that insulin-like growth factor-1 and insulin-like growth factor-2 mRNA levels were higher in tumor tissues and tumor-adjacent tissues than those in control cells, and were inversely correlated with the malignancy of the tumor with a positive correlation with ERα and ERß expression. Insulin-like growth factor-1R protein expression was correlated with clinical stage, and insulin-like growth factor-2R protein expression was inversely correlated with histological grade. CONCLUSIONS: Insulin-like growth factor system plays an important role in estrogen-induced endometrial carcinogenesis, and overexpression of insulin-like growth factor-1R in the advanced endometrioid adenocarcinoma is not estrogen-dependent.

[Inhibiting effect of letrozole combined with curcumin on xenografted endometrial carcinoma growth in nude mice].

BACKGROUND AND OBJECTIVE: Letrozole is an aromatase inhibitor that is used in the treatment of estrogen-sensitive tumors such as endometrial carcinoma, however, its therapeutic effect is still to be further improved. It is reported that curcumin has antitumor capability and can enhance the sensitivity of tumor cells to anticancer agents. This study was to investigate the inhibitory effect of letrozole combination with curcumin on the implanted endometrial tumor growth. METHODS: Nude mice were implanted with endometrial carcinoma RL-952 cells. All tumor-bearing mice were randomly divided into 5 groups: control(without treatment), Let(1) (letrozole, 1 microg/d), Let(10) (letrozole, 10 microg/d), Cur [ curcumin, 300 mg/kg.d)], and Let + Cur group [10microg/d letrozole + 50mg/ (kg.d) curcumin]. The tumor growth was monitored. Tumor cells apoptosis was detected in both control and treated groups. The expressions of bcl-2 mRNA and bcl-2 protein were detected using RT-PCR and Western blot, respectively. RESULTS: Fifty mice were successfully implanted with the endometrial tumor. Treatment with letrozole markedly inhibited tumor growth; the inhibitory effect was further enhanced by the combination of letrozole and curcumin. The inhibitory rates in Let (1), Let (10), the Cur, and the Let + Cur groups were 15.95%, 22.49%, 21.57%, and 35.89%, respectively. Treatment with curcumin inhibited the expression of bcl-2 in tumor cells at the mRNA and protein levels. The apoptosis rates in the control group and the four experimental groups mentioned above were 16.97%, 32.90%, 35.80%, 34.16%, and 47.24%, respectively. Tumor cells apoptosis were observed in mice treated with either letrozole or curcumin; however, combination of letrozole and curcumin further enhanced the inhibitory rate in tumor growth. CONCLUSIONS: Treatment with letrozole or curcumin could inhibit the xenografted endometrial tumor growth by inducing apoptosis in tumor cells. Combination of letrozole and curcumin further enhanced the inhibitory effect of tumor growth.

Uterus-like mass in the left broad ligament misdiagnosed as a malformation of the uterus: a case report of a rare condition and review of the literature.

OBJECTIVE: To describe a rare extrauterine mass of an unknown origin that was composed of smooth muscle with a central cavity lined by endometrium, which resembled a uterus. DESIGN: Case report. SETTING: Academic teaching hospital. PATIENT(S): A 17-year-old girl with severe dysmenorrhea was found to have a uterus-like mass composed of smooth muscle and lined by endometrium in the left broad ligament. INTERVENTION(S): Abdominal surgery. RESULT(S): The mass was successfully removed surgically, and her dysmenorrhea resolved. Histopathological examination revealed that the uterus-like mass was composed of smooth muscle with a cavity lined by endometrial glands and stroma. CONCLUSION(S): The histogenesis of a uterus-like mass is theorized to be a Müllerian system anomaly, metaplastic change, and/or heterotopia. We believe metaplasia is the most likely cause in this case. Alhough rare, it should be considered in the differential diagnosis of dysmenorrhea of uncertain etiology.

Aromatase inhibitor letrozole in synergy with curcumin in the inhibition of xenografted endometrial carcinoma growth.

OBJECTIVE: Endometrial carcinoma is a malignant tumor in the uterus. Its current treatment is not satisfactory. The present study aimed to promote the inhibitory effect on the implanted endometrial tumor growth. METHODS: Nude mice were implanted with endometrial carcinoma. Some tumor-laden mice were treated with aromatase inhibitor letrozole and/or curcumin. The tumor growth was monitored. Tumor cell apoptosis was detected in both control and treated groups. RESULTS: Fifty mice were successfully implanted with the endometrial tumor. Treatment with letrozole markedly inhibited the tumor growth; the inhibitor effect was further strengthened by combination with letrozole and curcumin. The results also showed that letrozole enhanced the expression of Bax and cytochrome c release and suppressed the expression of estrogen receptor in tumor cells. Treatment with curcumin inhibited the expression of Bcl-2 in tumor cells at the mRNA and protein levels. Tumor cell apoptosis was observed in mice treated with either letrozole or curcumin; however, combination of letrozole and curcumin further enhanced the inhibitory rate in tumor growth. CONCLUSIONS: Treatment with either letrozole or curcumin could inhibit the xenografted endometrial tumor growth via inducing apoptosis in tumor cells. Combination of letrozole and curcumin further strengthened the inhibitory effect on tumor growth.

[Angiogenesis and micrometastases of regional lymph node in ovarian cancer].

BACKGROUND & OBJECTIVE: In many investigation it was demonstrated that intratumor microvessel density (IMD) is the best factor for no tumor survival for patients with ovarian cancer. Occult axillary node micrometastases as the prognostic factor for the patients with breast cancer was proved, but currently no literature of the pelvic lymph node micrometastases in ovarian cancer is provided. This study try to detect the microvessel density of tumor tissues and micrometastases of regional lymph nodes in ovarian cancer and to investigate the relationship between these two factors and their prognostic significance. METHODS: The sections of ovarian cancer from 39 patients were stained immunohistochemically for cytokeratin(CK) and factor VIII-related antigen (F8-RA). Microvessels on the section were counted by computerized morphometry (40 fields). Sections of 212 regional lymph nodes from 39 patients were stained also with HE and CK. RESULTS: (1)We found 7 lymph node metastases with HE staining in 39 patients and 12 micrometastases with CK staining.(2) Average value of intratumor microvessel density (IMD) in the HE positive group was 48.86+/-16.60 and was obviously higher than that in the HE negative group (29.16+/-10.02,P< 0.01). The average IMD was 41.67+/-21.69 in the CK positive group and was also obviously higher than that in the CK negative group (28.70+/-10.77,P< 0.05). We found 8(50%) micrometastases in 16 patients with higher IMD (> or = 30/40 x fields) and only 4(17.4%) micrometastases in 23 patients with lower IMD (< 30/40 x fields), the difference was significant (P< 0.05). (3)IMD (P=0.03) and regional lymph node micrometastases(P=0.04) were the most significant factors to the survival time through Cox proportional hezard model analysis,the IMD (P=0.0008) and clinical stage (P=0.03) were also the most important factors to predict recurrence. CONCLUSION: The study suggests the CK immunohistochemical staining could detect the micrometastases in the HE negative lymph node in patients with epithelial ovarian cancer. The IMD is closely related to the micrometastases and these two factors have prognostic significance in ovarian cancer. The IMD and micrometastases could act as independent prognosis factors of patients with ovarian cancer.