International Journal of Fertility and Sterility

246-251

Authors: Muhammad Haddad, Rafat Esmail, Homayoun Khazali

Monosodium glutamate (MSG) is a flavour enhancer that is used as a food additive (E621) in many parts of the world, especially in East Asian countries. However, in recent studies, it has been used as a neurotoxin because MSG is reported to cause neural degeneration in the hypothalamic arcuate of neonatal animals. The results of several studies show the negative effects of MSG injections on different parts of the hypothalamic-pituitary-gonadal (HPG) axis, in addition to its ability to inhibit secretion many reproductive neuropeptides, neurotrophic factors, and hormones, all of which play vital roles in the regulation of reproductive function. Oral administration or injection of large quantities of MSG into newborn animals results in a decrease in or overabundance of the production of many regulatory peptides of the male and female reproductive systems. In this review, we summarize the results of the most important studies that have examined the effect of oral consumption or injection of MSG on regulatory peptides of the HPG axis.

258-262

Authors: Mahdieh Nikfarjam, Leila Rashki Ghaleno, Abdolhossein Shahverdi, Shahab Mirshahvaladi, Seyed Mehdi Ghoreishi, Alireza Alizadeh Moghadam Masouleh

Background: Pomegranate is an ancient fruit containing Punicalagin, which has known as an effective antioxidant. Pomegranate peel was recognized as a phenol and tannin source, and pomegranate seed contains unique fatty acid (Punicic acid). Limited information exists about the influences of pomegranate peel and seed on antioxidant enzymes and proteins in the male reproduction system. This study was performed to determine the pomegranate peel and seed effects on the expression of antioxidant genes and DJ-1 protein in ram’s testis. Materials and Methods: In this experimental study, twenty-one mature Iranian rams were randomly divided into three groups (n=7 in each group), and fed experimental diets consisted of a control diet (C), a diet containing dry pomegranate seed pulp (S), and a diet containing pomegranate peel (P) for 80 days. All rams were offered isoenergetic and isonitrogenous rations. Testicular tissue samples were collected, and expression of Gpx1, Gpx4, Prdx4, Prdx5, and Sod2 genes was quantified by real-time polymerase chain reaction (RT-PCR). In addition, western blotting was used to evaluate DJ-1 expression at the protein level. Results: Gpx1 and Sod2 mRNA levels in the peel group were significantly (P<0.05) higher than control. Prdx5 mRNA level was increased (P<0.05) in the seeds group than in the control group. Gpx4 and Prdx4 expression were statistically not affected significantly by the experimental diet. Data analysis showed a significant (P<0.05) increase (1.5-fold) in the expression level of DJ-1 in peel groups than in control. Conclusion: The expression of antioxidant genes and DJ-1 protein in ram testes are more influenced by pomegranate peel than seed.

269-274

Authors: Risk Factors Associated with Recurrent Pregnancy Loss and Outcome of Pre-Implantation Genetic Screening of Affected Couples

Background: Recurrent pregnancy loss (RPL) is a multifactorial disorder which affects up to 5% of couples around the world. Several factors are considered to be involved in RPL; but, the etiology remains unexplained in 35-60% of cases. The aim of this study was to assess the frequency of risk factors associated with RPL in a group of our clinic clients, and their pre-implantation genetic screening (PGS) outcome. Materials and Methods: We designed a retrospective descriptive study among, 602 Iranian couples referred to the Royan Reproductive Clinic (Tehran-Iran) from 2006 to 2018. Their karyotyping test and PGS outcomes were analyzed. PGS had been applied by array comparative genomic hybridization (array-CGH) on embryos from these patients. Also, karyotyping test had been performed using standard cytogenetic techniques. Results: G-banding analysis revealed a frequency of 15.61% chromosomal abnormalities in RPL couples. Also, the reciprocal translocations were more frequent (33/1204 cases) compared to the other structural abnormalities. Pregnancy rate per embryo transferred were 50% with array-CGH approach. Conclusion: Our findings could confirm a positive correlation between chromosomal abnormalities and RPL rate. Applying PGS for the RPL couples, leads to improvement of pregnancy success rate.

280-285

Authors: Ghazal Neysanian, Mahboube Taebi, Atefeh Rezaeian, Mohammad Hosein Nasr-Esfahani, Maryam Jahangirifar

Background: Based on studies on animal models, vitamin D plays an essential role in reproduction by controlling Ca and Mg levels. Despite these findings, the effects of vitamin D deficiency and supplementation on the outcome of assisted reproductive techniques (ART) remain controversial. Therefore, the aim of the present study was to assess the relationship between serum and follicular fluid 25-OH vitamin D levels on reproductive outcomes of infertile women. Materials and Methods: This prospective cohort study included 150 infertile women who underwent in vitro fertilization(IVF) or intracytoplasmic sperm injection (ICSI). The participants were allocated to one of the three groups according to their serum and follicular fluid 25-OH vitamin D concentrations (less than 10 ng/ml, between 10 and 30 ng/ml and more than 30 ng/ml), and fertilization, cleavage and biochemical and clinical pregnancy rates were compared among the groups. Data was analyzed by SPSS software and using Chi-square and Spearman correlation coefficient. Results: Serum and follicular fluid vitamin D levels significantly correlated with biochemical (P=0.008), (P=0.003) and clinical pregnancy (P=0.017), (P=0.001) rates respectively . However, the quality of embryos (P=0.125), (P=0.106)and fertilization rate (P=0.082), (P=0.059) were not associated with the level of serum and follicular fluid vitamin D. Conclusion: This study found that women with higher levels of vitamin D in their serum and follicular fluid are significantly more likely to achieve pregnancy but without affecting the quality of embryo and fertility rate.

294-299

Authors: Bahar Shakerian, Engin Turkgeldi, Sebile Guler Cekic, Sule Yildiz, İpek Keles, Baris Ata

Background: This study compared outcomes of the standard 6000 IU human chorionic gonadotropin (hCG) trigger with a dual trigger comprised of 6000 IU hCG and 1 mg leuprolide acetate for final oocyte maturation in an intracytoplasmic sperm injection (ICSI) cycle. By convention, ICSI was performed in most cases at the clinic. Materials and Methods: In this retrospective study, a total of 50 women were included in each arm. Participants were matched for age, indication and number of prior assisted reproduction technology (ART) cycles. Women at risk for ovarian hyperstimulation syndrome (OHSS) were excluded. A flexible gonadotropin releasing hormone (GnRH) antagonist protocol was used and final oocyte maturation was triggered when two leading follicles were >17 mm. Distribution of variables was evaluated visually with histograms. Continuous variables were defined by mean (standard deviation) or median (25th-75th percentile) depending on distribution characteristics. Categorical variables were defined by numbers and percentages. Continuous variables were compared between the groups with the t test or Mann-Whitney U test as appropriate. Categorical variables were compared by the chi-square test and its derivatives as appropriate. A two-sided P<0.05 indicated statistical significance. Results: Both groups had similar antral follicle counts, median parity (0) and number of previous failed cycles (0). The median number of oocytes (8 vs. 7), metaphase-two oocytes (6 vs. 5.5), blastocysts (1 vs. 1), clinical pregnancy rates (CPR) (28% vs. 22%), ongoing pregnancy rates (OPR) (22% vs. 20%) and pregnancy rate per transfer (53.3% vs 53.8%) were similar between the dual trigger and hCG only groups, respectively. Conclusion: Dual trigger for oocyte maturation stimulation failed to improve the ICSI outcome.

303-304

Authors: Koray Görkem Saçıntı, Yavuz Emre Şükür, Murat Sönmezer, Cem Somer Atabekoğlu

Although ovarian tissue cryopreservation is still considered as an experimental technique, several authors from around the world have reported successful and promising results. Currently, oocyte cryopreservation seems to be the most feasible technique for fertility preservation when there’s some kind of a time constraint in adolescents and adults. However, it has been estimated that a young woman would be expected to have a 94% likelihood of having a live birth with 20 mature frozen oocytes (1). At age 34 years, however, this expectation is decreased to 90% with 20 mature frozen oocytes. In addition to age-related limitations, an immediate obstacle for obtaining oocytes in cancer patients is the fact that only one controlled ovarian hyperstimulation (COH) cycle can usually be performed in these women because of time constraints, yielding a relatively low number of oocytes and/or embryos. For this reason, results from egg donation programs cannot be extrapolated to cancer patients, nor can the quality of oocytes be guaranteed. Hence, a combined fertility preservation technique can be of valuable in increasing the chances of successful future pregnancies following gonadotoxic cancer therapies. Previously, Dolmans et al. (2) suggested that cryopreservation of bilateral ovarian cortex followed by COH is a feasible and safe approach to preserve fertility before gonadotoxic treatment, and that the number of cryopreserved embryos was similar to the controls. We have been offering the option of the combined technique to fertility preservation patients for a couple of years and have performed it in a series of eight candidate patients. All patients had enough time for COH before oncology treatments. We first performed laparoscopic ovarian resection for ovarian tissue cryopreservation and then started COH on postoperative day 0 or 1 in each patient (Table 1). The main point in our findings is that ovarian resection is performed from the side with less antral follicle count of the patients. We suggest that this approach can increase the oocyte yield in a single available COH cycle. The data is limited on the effectiveness of combined technique and more long-term follow-up studies are needed in larger groups with appropriate controls. According to our clinical experience, we believe that combined technique is a valid approach, which is expanding beyond the experimental stage and has become a clinical technique for fertility preservation. We particularly suggest selecting the ovary with a low antral follicle count for wedge resection to increase oocyte yield. The information gathered from large international multicenter reports would encourage physicians to agree that the method should complete the experimental phase and be ready for wider clinical use in female fertility preservation.

167-177

Authors: Cyrus Jalili; Fuzieh Khani Hemmatabadi; Mehrdad Bakhtiyari; Amir Abdolmaleki; Fatemeh Moradi

Background: Prior to chemotherapy interventions, in vitro maturation (IVM) of folliclesthrough vitrification can be used to help young people conserve their fertility. Materials and Methods: This experimental study was conducted on immature female BALB/c mice (12-14 days). Follicles were gathered mechanically and placed in α-Minimal Essential Medium (α-MEM) containing 5% fetal bovine serum (FBS). Some pre-antral follicles were frozen. The fresh and vitrified follicles were cultured in different concentrations of sodium alginate (0.25%, 0.5%, and 1%) and two dimensional (2D) medium for 12 days. The samples were evaluated for viability percentage, the number of MII-phase oocytes and reactive oxygen specious (ROS) level. Additionally, Gdf9, Bmp15, Bmp7, Bmp4, Gpx, mnSOD and Gcs gene expressions were assessed in the samples. Results: The highest and lowest percentages of follicle viability and maturation in the fresh and vitrified groups were respectively 0.5% concentration and 2D culture. There was no significant difference among the concentrations of 0.25% and 1%. Viability and maturation of follicles showed a significant increase in the fresh groups in comparison with the vitrified groups. ROS levels in the both fresh and vitrified groups with different concentrations of alginate showed a significant decrease compared to the control group. ROS levels in follicles showed a significant decrease in the fresh groups in comparison with the vitrified groups (P≤0.0001). The highest gene expression levels were observed in the 0.5% alginate (P≤0.0001). Moreover, the viability percentage, follicle maturation, and gene expression levels were higher in the fresh groupsthan the vitrified groups (P≤0.0001). Conclusion: Alginate hydrogel at a proper concentration of 5%, not only helps follicle get mature, but also promotes the expression of developmental genes and reducesthe level of intracellular ROS. Follicular vitrification decreases quality of the follicles, which are partially compensated using a three dimensional (3D) cell culture medium.

189-196

Authors: Mahboubeh Taebi; Nourossadat Kariman; Ali Montazeri; Hamid Alavi Majd

Background: Infertility stigma is a phenomenon associated with various psychological and social tensions especially for women. The stigma is associated with a feeling of shame and secrecy. The present study was aimed to explore the concept of infertility stigma based on the experiences and perceptions of infertile women. Materials and Methods: This qualitative conventional content analysis study was conducted in Isfahan Fertility and Infertility Center, Iran. Data were collected through in-depth interviews with 17 women who had primary infertility. All the interviews were recorded, transcribed and analyzed according to the steps suggested by Graneheim and Lundman. The Standards for Reporting Qualitative Research (SRQR) checklist was followed for this research. Results: Eight hundred thirty-six initial codes were extracted from the interviews and divided into 25 sub-categories, 10 categories, and four themes. The themes included “stigma profile, self-stigma, defensive mechanism and balancing”. Stigma profile was perceived in the form of verbal, social and same sex stigma. Self-stigma was experienced as negative feelings and devaluation. Defensive mechanism was formed from three categories of escaping from the stigma, acceptance and infertility behind the mask. Two categories; empowered women and pressure levers, created a balancing theme against the infertility stigma. Conclusion: Infertile women face social and self-stigma which threatenstheir psychosocial wellbeing and self-esteem. They use defensive response mechanisms and social support to mitigate these effects. Education focused on coping strategies might be helpful against infertility stigma.

202-209

Authors: saeed Azizollahi; Maryam Bagheri; Fedyeh Haghollahi; Seyyede Momeneh Mohamadi; Batool Hosein Rashidi

Background: Gonadotropin-releasing hormone (GnRH) analogues have been extensively utilized in the ovarian stimulation cycle for suppression of endogenous rapid enhancement of luteinizing hormone (LH surge). Exclusive properties and functional mechanisms of GnRH analogues in in vitro fertilization (IVF) cycles are clearly described. This randomized clinical trial was performed to evaluate clinical and molecular impacts of the GnRH agonist and antagonist protocols in IVF cycles. For this purpose, gene expression of cumulus cells (CCs) as well as clinical and embryological parameters were evaluated and compared between two groups (GnRH agonist and antagonist) during the IVF cycle. Materials and Methods: Twenty-one infertile individuals were enrolled in this double-blind randomized clinical study. Subjects were randomly allocated into two groups of GnRH agonist (n=10) treated patients and GnRH antagonist (n=11) treated individuals. The defined clinical embryological parameters were compared between the two groups. Expression of BAX, BCL-2, SURVIVIN, ALCAM, and VCAN genes were assessed in the CCs of the participants using the real-time polymerase chain reaction (PCR) technique. Results: The mean number of cumulus oocyte complex (COC), percentage of metaphase II (MII) oocytes, grade A embryo and clinical parameters did not show noticeable differences between the two groups. BAX gene expression in the CCs of the group treated with GnRH agonist was remarkably higher than those received GnRH antagonist treatment (p <0.001). The mRNA expression of BCL-2 and ALCM genes were considerably greater in the CCs of patients who underwent antagonist protocol in comparison to the group that received agonist protocol (p <0.001). Conclusion: Despite no considerable difference in the oocyte quality, embryo development, and clinical outcomes between the group treated with GnRH agonist and the one treated with antagonist protocol, the GnRH antagonist protocol was slightly more favorable. However, further clinical studies using molecular assessments are required to elucidate this controversial subject (Registration number: IRCT20101115005181N16).

219-225

Authors: Induja Manikandan; Sushmita Bora; Prashant Adole; Chitra Thyagaraju; Rajesh Ganesh

Background: Because of the widespread use of organophosphate (OP) pesticides in agriculture, they are major environmental contaminants in developing countries. OP pesticides decrease sperm concentration and affect its quality, viability, and motility. Studies have demonstrated the association between abnormal semen analysis and OP pesticides exposure among the high-risk population. Asthere is limited data on the percentage of OP pesticides exposure, the study aimed to determine the OP pesticides exposure in Southern Indian men with idiopathic abnormal semen analysis and find the possible source of their OP pesticides exposure. Materials and Methods: In this cross-sectional pilot study, fifty men with idiopathic abnormal semen analysis as cases and fifty men with normal semen analysis as controls were recruited. Detailed history wastaken and general and systemic examinations were carried out. OP pesticides exposure was determined by assessment of pseudocholinesterase and acetylcholinesterase levels and urinary OP pesticides metabolites dialkyl phosphate (DAP) consisting of dimethyl phosphate (DMP), diethyl thiophosphate (DETP), and diethyl dithiophosphate (DEDTP). Results: Cases had statistically significantly lower levels of pseudocholinesterase (5792.07 ± 1969.89 vs. 10267.01 ± 3258.58 IU/L) (P=0.006) and acetylcholinesterase [102.90 (45.88-262.74) vs. 570.31 (200.24-975.30) IU/L] (P=0.001) as compared to controls. Cases had a statistically significantly higher percentage of urinary DAP positivity as compared to controls (80 vs. 38%) (p <0.0001). Hence, cases had a significantly higher percentage of OP pesticides exposure as compared to controls (20 vs. 4 %) (P=0.015). OP-exposed cases had significantly higher urinary DETP and DEDTP levels as compared to OP non-exposed cases. Also, urinary DETP and DEDTP levels were significantly negatively associated with sperm concentration, motility, and normal morphology among OP-exposed cases. Conclusion: Southern Indian men with idiopathic abnormal semen analysis had a significantly higher percentage of OP pesticides exposure as compared to men with a normal semen analysis.

234-240

Authors: Arezoo Bagheri; Mahsa Saadati

Background: Total fertility rate (TFR) in Iran decreased from the year 2000 and recently Iran has experienced fertility rates below replacement level. Birth interval is one of the most important determinants of fertility levels and plays a vital role in population growth rate. Due to the importance of this subject, the aim of this study was analyzing three birth intervals using three Survival Recurrent Event (SRE) models. Materials and Methods: In a 2017 cross-sectional fertility survey in Tehran, 610 married women, age 15-49 years, were selected by multi-stage stratified random sampling and interviewed using a structured questionnaire. The effects of selected covariates on first, second and third birth intervals were fitted to the data using the Prentice-Williams- Peterson-Gap Time (PWP-GT) SRE model in SAS 9.4. Results: Calendar-period had a significant effect on all three birth intervals (p <0.01). The Hazard Rate (HR) for a short birth interval for women in the most recent calendar-period (2007-2017) was lower than for the other calendarperiods. Women’s migration influenced second (P=0.044) and third birth intervals (P=0.031). The HR for both birth intervals in migrant women was 1.298 and 1.404 times shorter, respectively than non-migrant women. Women’s employment (P=0.008) and place of residence (p <0.05) also had significant effects on second birth interval; employed women and those living in developed, completely-developed and semi-developed areas, compared to unemployed women and those living in developing regions, had longer second birth intervals. Older age at marriage age increased the HR for a short third birth interval (p <0.01). Conclusion: The analysis of birth interval patterns using an appropriate statistical method provides important information for health policymakers. Based on the results of this study, younger women delayed their childbearing more than older women. Migrant women, unemployed women and women who live in developing regions gave birth to their second child sooner than non-migrant employed women, and women who lived in more developed regions. The implementation of policies which change the economic and social conditions of families could prevent increasing birth intervals and influence the fertility rate.

8-14

Authors: Rezvan Bahmyari, , Ali Ariafar, Mehrab Sayadi,Shirzad Hossieni, Sara Azima

Background: Male infertility may originate from a wide spectrum of conditions while in 30-40 percent of cases, no significant reason can be identified. Thereby, it is recognized as male idiopathic infertility. This study was undertaken to investigate the effect of daily intake of selenium, vitamin E and folic acid on sperm parameters in males with idiopathic infertility. Materials and Methods: Seventy infertile men were selected to participate in this single-blind, randomized controlled clinical trial using convenience sampling. They were equally divided into two groups via permuted block randomization method. The intervention group received selenium tablet (200 μg per day, oral), vitamin E capsule (400 IU per day, oral) and folic acid tablet (5 mg per day, oral).The placebo group received matching placebo for three months. Semen volume, total sperm motility, sperm concentration, progressive sperm motility, normal sperm morphology, sperm motility index (SMI) and functional sperm concentration (FSC) were assessed by sperm quality analyzer-v (SQAV) before and after the intervention. Paired t test, and independent t test were used to compare the results within and between the groups, respectively. The IBM SPSS V.16.005 was used for data analysis. A P<0.05 was considered statistically significant. Results: After three months, according to within-group analysis, a significant difference was found in mean SMI (P=0.007) and FSC (P=0.001) in the intervention group. According to between-group analysis, no significant difference was found in mean semen volume (P=0.610), sperm concentration (P=0.126), total sperm motility (P=0.765), progressive sperm motility (P=0.767), normal sperm morph (P=0.403), SMI (P=0.556) or FSC (P=0.706) between the groups. Conclusion: Consumption of selenium, vitamin E and folic acid in infertile men with asthenozoospermia was not effective (Registration number: IRCT2017012432153N1).

20-25

Authors: Mozhgan Raigani, Niknam Lakpour, Meysam Soleimani, Behrooz Johari, Mohammad Reza Sadeghi

Background: The relation between key enzymes in regulation of folate metabolism and male infertility is the subject of numerous studies. We aimed to determine whether 5, 10-methylenetetrahydrofolate reductase (MTHFR) C677T and methionine synthase reductase (MTRR) A66G genotypes are associated with male infertility in Iranian men and to evaluate its effect on seminal levels of folate and vitamin B12. Materials and Methods: In this retrospective study, semen and peripheral blood samples were collected from 254 men with oligoasthenoteratozoospermia (OAT) and 77 normozoospermic men who attended Avicenna infertility clinic. Single nucleotide polymorphism (SNP) analysis was carried out in genomic DNA by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method for MTHFR C677T and MTRR A66G gene polymorphisms. Results: In MTHFR C677T, our founding showed that T carrier was conversely lower in OAT than normozoospermic men (χ2-test=7.245, P=0.02) whereas in MTRR A66G, A and G carrier showed no significant difference between the two groups (χ2-test=1.079, P=0.53). The concentration of seminal folate was not different between normozoospermic (18.83 ± 17.1 ng/ ml) and OAT (16.96 ± 14.2 ng/ml) men (P=0.47). The concentration of vitamin B12 was slightly higher in normozospermic men (522.6 ± 388.1 pg/ml) compared to OAT men (412.9 ± 303.6 pg/ml, P=0.058). Conclusion: The MTHFR C677T and MTRR A66G have no effect on the concentrations of seminal folate and vitamin B12. The present study showed that two SNPs of MTRR A66G and MTHFR C677T cannot be seen as a risk factor for male factor subfertility.

34-39

Authors: Reiko Shiba; Masayuki Kinutani; Shinichiro Okano; Maki Ikeda; Emi Fukunaga; Yoshihisa Harada; Reo Kawano; Yuko Kikkawa

Background: Luteal phase support (LPS) is essential for hormone replacement therapy (HRT) for frozen-thawed embryo transfer (FET). However, the optimal dose and serum progesterone (P4) levels required for pregnancy are controversial. We attempted to determine the association between pregnancy outcomes and serum P4 levels administered via vaginal suppository for HRT-FET cycles on embryo transfer day. Materials and Methods: This was a secondary analysis of the dataset from the EXCULL trial, which prospectively investigated pregnancy outcomes of four different P4 vaginal suppositories (Lutinus, Utrogestan, Luteum, and Crinone) for HRT-FET. It was conducted at a private fertility clinic between December 2016 to December 2017. During this trial, 235 cycles were divided into four groups based on serum P4 values (quartile [Q] 1 group: <7.8 ng/mL; Q2 group: 7.8-10.8 ng/mL; Q3 group: 10.8-13.7 ng/mL; Q4 group: >13.7 ng/mL). We investigated clinical pregnancy rate (CPR), positive fetal heart rate (FHR), live birth rate (LBR), and miscarriage rate (MR) for each group. A logistic regression analysis was performed using age, body mass index (BMI), and transferred embryos as covariates. Results: Serum P4 values (ng/mL) of each drug were as follows: Lutinus, 13.3 ± 4.9; Utrogestan, 9.3 ± 3.3; Luteum, 13.6 ± 4.2; and Crinone, 8.7 ± 3.2 (mean ± standard deviation, P<0.001).The percentages of Utrogestan and Crinone were higher in the Q1 group, while the percentages of Lutinus and Luteum were higher in the Q4 group. Nonetheless, there were no statistical differences between the Q1 and Q4 groups in CPR, FHR, LBR, and MR. Conclusion: When vaginal P4 was used for FET, although serum P4 levels on transfer day differed based on the drug that was administered, no relationship was observed between serum progesteronelevels and pregnancy outcomes (Registration number: UMIN000032997).

44-50

Authors: Shabnam Shirazi; Bahram Pourghassem Gargari; Azimeh Izadi; Shiva Taghizadeh; Marziyeh Parizad

Background: Angiogenesis disturbances are common in women with polycystic ovary syndrome (PCOS). Vitamin E has antiangiogenic properties. Data on the effects of vitamin E on angiogenesis in PCOS is limited, so the current study was conducted to evaluate its effects on angiogenic indices in PCOS patients. Materials and Methods: This randomized, double-blind, placebo-controlled trial was performed on 43 women aged 20-40 years, diagnosed with PCOS (Rotterdam criteria). It was performed at the referral clinic affiliated to Tabriz University of Medical Sciences, Tabriz, Iran, from April 2017 to September 2017. Patients were randomly assigned into two groups to receive either 400 IU/day vitamin E -as alpha tocopheryl acetate- (n=22) or placebo (n=21), for 8 weeks. Anthropometric, and angiogenic parameters including body weight, fat mass and fat free mass, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin-1 (Ang- 1), and angiopoietin-2 (Ang-2) were measured by standard methods at the beginning and at the end of study. Statistical Package for Social Science version 25 was used for statistical analysis and P<0.05 were considered significant. Results: After adjusting for potential confounders, we observed that vitamin E supplementation significantly reduced body weight, fat mass, Ang-1, Ang-1/Ang-2 ratio and VEGF (P<0.01). We did not observe any considerable effect for vitamin E on Ang-2 level or bFGF. Conclusion: Vitamin E supplementation for 8 weeks in the PCOS women had beneficial effects on body weight, Ang- 1, Ang-1/Ang-2 ratio, and VEGF level (Registration number: IRCT201610193140N18).

60-64

Authors: Hajar Lotfollahi; Hedyeh Riazi; Reza Omani Samani; Saman Maroufizadeh; Ali Montazeri

Background: Sexual self-concept has a considerable impact on mental and sexual health. However, the relationship between sexual self-concept and infertility is unknown. This study aimed to compare sexual self-concept between fertile and infertile women. Materials and Methods: This cross-sectional study was conducted on a sample of 250 fertile and 250 infertile women who had referred to 9 health centers affiliated to Medical universities in Tehran and Royan infertility treatment clinics in Tehran, Iran in 2017. Sexual self-concept was measured using the Multidimensional Sexual Self-Concept Questionnaire (MSSCQ) consisting of 20 subscales. Analysis of covariance (ANCOVA) was performed to compare sexual self-concept between the two groups. Results: The mean age of fertile and infertile women was 34 ± 5.62 and 29.74 ± 5.29 years, respectively. The highest score in both groups was for the sexual self-schemata subscale (mean score for fertile=3.21 ± 0.68 and for infertile= 3.42 ± 0.62). The lowest score was for sexual-depression subscale (mean score for fertile=0.59 ± 0.81 and for infertile=0.61 ± 0.76). After adjustment for the age of each subject, the husband's age, duration of marriage, and women’s education, we analyzed the sexual-satisfaction, the power-other sexual control, and the fear-of-sex subscales, which were found to be significantly lower in infertile women (P<0.05). No other significant differences between the fertile and infertile groups were observed. Conclusion: We observed significant differences between fertile and infertile women in terms of sexual-satisfaction, the power-other sexual control, and the fear-of-sex, but not in other sexual self-concept subscales. These findings suggest that there is need to improve sexual self-concept among both fertile and infertile women. Indeed implementation of educational and counseling programs by reproductive health specialists might play an important role in enhancing sexual self-concept among these populations.

73-79

Authors: Mohammad Hossein Nasr-Esfahani; Khadijeh Ahmad khanbeigi; Ali Hasannia

Background: Human cloning is a recent occurrence that is not confined to bio-issues; rather, it has provoked numerous questions worldwide and presented scientific and religious challenges. These series of articles aim to examine the proposed approaches and analyze the aspects of human cloning in terms of tenets, morals, jurisprudence, and laws. In this paper, we analyze the ideological and theological evidences, regardless of scientific, ethical and legal problems that exist in the reproduction method. Materials and Methods: We used a descriptive-analytical method to consider the challenges of human cloning according to the “system of Divine creativity” and “the will of God”, as well as the “pairing system” and “diversity in nature” with emphasis on the Holy Qur’an and Qur’anic commentaries. Results: According to the Qur’an, although any type of physical changes and retouching of the human body are forbidden, the alteration of God's creation may not prove the prohibition of cloning. Cloning is not contradictory to the principle, precedent, and rule of coupling, and it may be one of the hidden precedents of creation. Therefore, not only does a clone not contradict the precedent of the variety of men, but this variety is a sign for men and not a precedent predominated over the order of nature. Conclusion: It is proven that cloning does not give life; instead, it utilizes the life bestowed by God. This technique does not contradict the precedents of existence. It is a way to discover some precedents of God and is under the order of cause and effect of the world. Cloning is not considered as a challenge to human beliefs, nor is it a change in Divine creation. Moreover, cloning does not contradict the theological teachings and concepts of the Holy Qur’an and Shiite Muslims.

171-175

Authors: Rahil Jannatifar; Kazem Parivar; Nasim Hayati Roodbari; Mohammad Hossein Nasr-Esfahani

Background: One of the important factor associated with male infertility is high production of reactive oxygen species (ROS). The main function of Nuclear factor erythroid 2-related factor 2 (NRF2) is to activate the cellular antioxidant response by inducing the transcription of a wide array of genes that can combat the harmful effects of factors such as oxidative stress. The purpose of this study was to evaluate the effect of N-acetyl-L-cysteine (NAC), as an antioxidant drug, on NRF2 Gene Expression in Asthenoteratozoospermia Men. Materials and Methods: In this randomized, blinded clinical trial study, included 50 infertile men with asthenoteratozoospermia, who received NAC (600 mg, three times daily). Sperm parameters analyzed according to the world health organization (WHO; 2010). Sperm DNA fragmentation, relative NRF2 expression, and seminal plasma level of antioxidant enzymes were measured by TUNEL assay, reverse transcription polymerase chain reaction (RT-PCR) and ELISA test, respectively. Results: After NAC treatment, findings showed a significant increase in sperm concentration and motility compared to pre-treatment status, whereas the percentage of abnormal morphology and DNA fragmentation was significantly decreased (P<0.05). A significant improvement in expression of NRF2 gene and antioxidant enzyme levels were observed compared to pre-treatment by NAC (P<0.05). Significant correlations were observed between NRF2 mRNA expression level, specific sperm parameters and level of antioxidant enzymes (P<0.05). Conclusion: The results demonstrated that NAC oral supplementation protected against oxidative stress by enhancing NRF2 expression. This could improve semen parameters quality parameters in asthenoteratozoospermia men (Registration number: IRCT20170830035998N4).

185-192

Authors: Masha Shirani; Praveen Saneei; Mehran Nouri; Mohamadreza Maracy; Homayoun Abbasi; Gholamreza Askari

Background: This cross-sectional study pointed to assess the relationship between major dietary patterns and dietary diversity score with semen parameters, in infertile Iranian males. Materials and Methods: In this cross-sectional study, 260 infertile men (18-55 years old) who met the inclusion criteria, entered the study. Four Semen parameters, namely sperm concentration (SC), total sperm movement (TSM), normal sperm morphology (NSM) and sperm volume were considered according to spermogram. A 168-item food frequency questionnaire (FFQ) was used to collect dietary intakes and calculate dietary diversity score. Factor analysis was used to extract dietary patterns. Results: The following four factors were extracted: “traditional pattern”, “prudent pattern”, “vegetable-based pattern” and “mixed pattern”. After adjusting potential confounders, those in the highest quartile of the traditional pattern had 83% less odds for abnormal concentration, compared with the first quartile (OR=0.17, 95% CI: 0.04-00.73); however, subjects in the highest quartile of this pattern had 2.69 fold higher odds for abnormal sperm volume as compared with those of the first quartile (95%Cl: 1.06-6.82). Men in the second quartile of prudent pattern had 4.36 higher odds of an abnormal sperm volume in comparison to the reference category (95%CI: 1.75-10.86), after considering potential confounders. With regard to mixed pattern, men in the second, third and fourth quartile of this pattern had respectively 85 (5%Cl: 0.03-0.76,), 86 (95%Cl: 0.02-0.75) and 83 % (95%Cl: 0.034-0.9) less odds of abnormal concentration, compared with the first quartile. Additionally, no significant association was found between dietary diversity score and sperm quality parameters. Conclusion: Higher intake of the traditional diet was linked to lower abnormal semen concentration and poorer sperm volume. Also, the mixed diet was associated with reduced prevalence of abnormal semen concentration.

201-208

Authors: Fatemeh Pashizeh; Reza Mansouri; Fatemeh Davari-Tanha; Reyhaneh Hosseini; Zahra Asgari; Hamideh Aghaei; Farangis Najafi Arbastan; Samira Rajaei

Background: Endometriosis is a chronic inflammatory disorder with known immune disturbances. The aim of this study was to compare the frequency of different CD4+ T cells [T helper (Th)1, Th2, Th17 and regulatory T cells (Tregs)] in peripheral blood (PB) and peritoneal fluid (PF) of patients that have early and advanced stages of endometriosis with a control group. Materials and Methods: In this case control study, PB and PF samples were collected from women aged 24-40 years who underwent laparoscopy procedures. The frequency of CD4+ T subsets were analysed by flow cytometry and compared between three study groups; early endometriosis (stage I, II), advanced endometriosis (stage III, IV) and control (no endometriosis). T cell numbers were compared between the PB and PF in each of the aforementioned groups. Results: No statistically significant difference was found between the study groups regarding the numbers of Th1, Th2 and Th17 cells in PB. The PF of patients with advanced endometriosis had increased numbers of Th17 cells compared to the control group (P=0.003), with P values of 0.059 and 0.045 in both menstrual phases. Increased numbers of Th2 cells in PF from early compared to advanced stages of endometriosis were detected exclusively in the luteal phase (P=0.035). The control group had increased numbers of Treg and Th2 cells in the PF compared to PB (both, P value=0.046). However, in the early stages of endometriosis there were more Th2, Th17 and Treg cells in the PF compared to PB (P values: 0.005, 0.047 and 0.013, respectively), while the number of Th17 cells was higher in the PF compared with PB in the advanced stages of endometriosis (P= 0.013). Conclusion: There were increased numbers of Th17 cells in the PF of patients with advanced stages of endometriosis, which could be related to the severity of this disease.

213-217

Authors: Pinar Gulsen Coban; Ayla Sargin Oruc; Meryem Kuru Pekcan; Hasan Inal; Necati Hancerliogullari; Nafiye Yilmaz

Background: We investigated the impact of the choice of catheter type and tenaculum use on pregnancy related outcomes in intrauterine insemination (IUI) treatments. Materials and Methods: A total of 338 consecutive IUI cycles were assessed in this retrospective study. Participants were divided according to the insemination technique - soft catheter (group 1; n=175), firm catheter (group 2; n=100), or tenaculum (group 3; n=63). Clinical, laboratory, semen parameters and pregnancy related outcomes were compared. Results: Demographic characteristics and laboratory parameters were similar between the groups (P>0.05). The clinical pregnancy rate (CPR) was significantly higher in the firm catheter (19%, 19/100) and tenaculum (31.7%, 20/63) groups compared to the soft catheter group (5.1%, 9/175, P<0.001). There were no significant differences between the groups in live birth and miscarriage rates per clinical pregnancy (P>0.05). Conclusion: Our findings indicate that the use of a firm catheter or tenaculum for IUI might result in a higher CPR, but might not have a considerable effect on the live birth rate (LBR). Further prospective randomized studies are required to determine the long-term effects of the catheter type or tenaculum use on IUI success.

223-227

Authors: Nadia Hossein-Zadeh; Morteza Bagheri; Isa Abdi Rad; Marziyeh Lozeie; Mahdieh Nasir-Zadeh

Background: 3,4-Methylenedioxymethamphetamine (MDMA) disrupts function of the endocrine system and different organs such as heart, blood vessels, kidney, liver and nervous systems. This study was conducted to evaluate impact of MDMA on apoptosis and Zinc in the MDMA-induced apoptosis of cultured Sertoli cells by measuring Caspase-3 gene expression. Materials and Methods: In this experimental study, Sertoli cells were incubated with MDMA (0, 0.5, 1, 3, 5 mM), Zinc (0, 8, 16, 32, 64 μM) and Zinc (8 μM) prior to adding MDMA (5 mM) for 24 and 48 hours. MTT assay was used for evaluating impacts of these conditions on the viability of Sertoli cells. Caspase-3 gene expression level was detected using quantitative reverse transcription PCR (qRT-PCR) in all of the tested groups. Results: Finding showed that cellular viability was decreased and level of Caspase-3 mRNA was increased in MDMA treated cells. Additionally, pre-treatment with Zinc (8 μM) attenuated MDMA-induced apoptosis and down-regulated caspase-3. The mean of caspase-3 mRNA level (fold change ± SE) was 3.98 ± 1.18, 0.31 ± 0.28, and 1.72 ± 0.28 in respectively MDMA (5 mM), Zinc (8 μM), and Zinc+MDMA groups vs. control group. The mean of Caspase-3 mRNA (fold change) was not statistically different in the tested groups (P>0.05), unless MDMA (5 mM) group (P = 0.008). Conclusion: We suggest that MDMA toxicity could be involved in apoptosis of Sertoli cells. In addition, Zinc could reduce MDMA-induced apoptosis by down-regulation of Caspase-3 mRNA levels.

234-239

Authors: Azadeh Ghaheri; Reza Omani-Samani; Mahdi Sepidarkish; Mostafa Hosseini; Saman Maroufizadeh

Background: The most common mental disorders in infertile patients are depression and anxiety. The four-item Patient Health Questionnaire-4 (PHQ-4) is a widely used tool that consists of the PHQ-2 depression and Generalized Anxiety Disorder-2 (GAD-2) scales. Given that PHQ-4 has not been validated in infertile patients, this study aimed to examine its reliability and validity in this population. Materials and Methods: Participants in this cross-sectional study consisted of 539 infertile patients from a referral fertility centre in Tehran, Iran. The PHQ-4, Hospital Anxiety and Depression Scale (HADS), World Health Organisation- Five Well-Being Index (WHO-5), Penn State Worry Questionnaire (PSWQ) and demographic/infertility questionnaires were administered to all participants. Factor structure and internal consistency of PHQ-4 were evaluated via confirmatory factor analysis (CFA) and Cronbach’s alpha, respectively. The convergent validity of this scale was examined by its relationship with HADS, WHO-5 and PSWQ. Results: CFA results provided support for a two-factor model of PHQ-4. Internal consistency of the PHQ-4 and its subscales both were elevated with Cronbach’s alpha coefficients of 0.767 (PHQ-4), 0.780 (PHQ-2) and 0.814 (GAD-2). Inter-item correlations were between 0.386 and 0.639, and corrected item-total correlations were between 0.576 and 0.687. PHQ-4, PHQ-2 and GAD-2 showed positive correlations with measures of HADS-anxiety, HADS-depression, and PSWQ and negative correlations with WHO-5, which confirmed convergent validity. Among demographic/fertility variables, we observed that gender, infertility duration, and failure in previous treatment were correlated with PHQ-4 and its subscales scores. Conclusion: The PHQ-4 is a reliable and valid ultra-brief screening instrument for measuring both anxiety and depressive symptoms in infertile patients.

247-255

Authors: Mahsa Saadati; Arezoo Bagheri

Background: Birth spacing, especially the first birth interval (FBI), is a suitable index to investigate the delayed fertility that results in a low fertility pattern. Non-parametric familiar alternatives to the Cox proportional hazard regression (CPH) model include survival trees that can automatically discover certain types of covariate interactions according to the survival length. The aim of this research is to study FBI influential factors by applying survival trees. Materials and Methods: In this cross-sectional study, 610 married women (aged 15-49 years), were selected from different regions of Tehran, Iran in the Winter and Spring of 2017. Classification and regression trees (CART) for the FBI survival tree were fitted by taking into consideration the predictors of each woman’s age, age at first marriage, educational level, partner’s educational level, activity, region, house ownership, kinship, partner’s race, marriage time attitude, and expenditure using R packages. Results: Since the PH assumption of the CPH model was not confirmed for the covariates of age at first marriage (P=0.001), kinship (P=0.000), partner’s race (P=0.001), and marriage time attitude (P=0.042), the results of this model were not valid. Thus, a CART survival tree was fitted. The validity of the fitted model in assessing FBI was confirmed by the significant result of the log rank test (P<0.01) for the terminal nodes and the value of the separation measure, which was greater than 1. The fitted tree had 13 terminal nodes and the most vital FBI predictor was women’s age. The longest FBI belonged to educated and employed women, ages 30-37 years. Conclusion: Analysing patterns of birth spacing by selecting the appropriate statistical method provides important information for health policymakers. In order to formulate appropriate demographic policies, it is essential to take into consideration age, educational level and job status of the women, all of which have essential roles on their decision to have children.

260-261

Authors: Antonio Simone Laganà; Marco Scioscia

Endometrial carcinoma (EC) has a worldwide incidence of 8.4 per 100,000 persons per year and is a leading cause of cancer mortality in women in developed countries after ovarian and cervical cancer (1). Among the gynecological diseases, polycystic ovary syndrome and uterine leiomyomas have been associated with a potential increased risk to develop EC, whereas less robust data are available for endometriosis and adenomyosis (2). The association between EC and adenomyosis, defined by the presence of ectopic endometrial glands and stroma within the myometrium, is still debated: on the one hand, adenomyosis is found with an incidence of 10 to 18% in EC specimen at final histology after hysterectomy.

264-271

Authors: Reihaneh Pirjani; Maryam Rabiei; Ameneh Abiri; Ashraf Moini

In this article, we reviewed and compared some of COVID-19 and pregnancy guidelines; this can be useful for pregnant women including those with a history of infertility specially those undergone assisted reproductive techniques (ART). The general advice given for prenatal care is to reduce face-to-face visits. All women who refer for prenatal visits should be evaluated for signs of the infection at the time of entry. The triage of suspected women should be done separately from other patients. Outpatient monitoring with a 14-day selfquarantine can be considered for asymptomatic infected women and for those with mild symptoms. Inpatient management criteria include moderate to severe symptoms and the target level of oxygen saturation is 92 to 95% in different guidelines. In the presence of fever, it is important to conduct a thorough examination of other causes of the fever. It is important to monitor fluid intake and output, maintain fluid and electrolyte balance and prevent fluid overload. Thromboembolic prophylaxis is recommended. Corticosteroid administration is based on obstetrics indications, while in critical ill cases, it should be based on multi-disciplinary teams (MDT) decision. A positive COVID-19 result in the absence of other obstetrics causes, cannot be considered an indication for delivery in mild and asymptomatic cases. In critically ill pregnant women, an individualized decision should be made about delivery time by the MDT. General anesthetic should be avoided unless inevitable for standard procedures such as intubation is an aerosol-generating procedure (AGP). There is agreement on the point that babies born to infected mothers, even if isolated from the mother at birth, should be considered a close contact of the mother and tested for COVID-19 and separated from other neonates. Breastfeeding is encouraged and hand hygiene and face mask during feeding are highly recommended by all guidelines

84-90

Authors: Hamid Ekrami, Mansoureh Movahedin, Fereshteh Koosha, Zohreh Mazaheri, Manijhe Mokhtari-Dizaji

Objective The study aimed to evaluate the impact of Calligonum extract and US radiation on sperm parameters of cryopreserved human semen samples. Materials and methods In this experimental study, twenty-five semen specimens were obtained from healthy semen donors and incubated in human tubal fluid (HTF) medium supplemented with 10% human serum albumin (HSA) for 45 minutes. Samples were treated with Calligonum extract (10 μg/ml) alone (CGM group) and US radiation (LIPUSexposed group) alone or a combination of both treatments (CGM+LIPUS). The US group received US stimulation (in both continuous and pulsed wave modes) at a frequency of 1 MHZ and intensity of 200 mW/cm2 for 200 seconds. Sperm morphology was assessed by Diff-Quik staining. The DNA fragmentation was evaluated the Halo sperm kit. Sperm parameters was analyzed by a computer-assisted semen analysis system. Reactive oxygen species (ROS) was assessed by flow cytometry. Results The results showed that the treatment with Calligonum extract significantly (P<0.05) increased the progressive motility of spermatozoa in the CGM group as compared with the control group. The application of low-intensity US significantly (P<0.05) decreased the motility and viability of spermatozoa in the US group when compared with the control group. Our findings also indicated that the use of both low-intensity US in continuous mode and Calligonum extract slightly increased progressive motility; however, such an increase was not statistically significant. The rate of DNA fragmentation was considerably higher (P<0.05) in control and LIPUS-exposed groups than the other groups. Conclusion Treatment of spermatozoa with Calligonum extract slightly improved the sperm parameters due to its antioxidant activity, on the other hand, according to our results, US radiation did not improve sperm parameters which may be due to interference with the motility of sperm, as well as its physical effects on spermatozoa.

102-109

Authors: Ajit Kumar Saxena, Meenakshi Tiwari, Mukta Agarwal, Aprajita Aniket Kumar

Objective The present study has been designed with the aim of evaluating A-kinase anchoring proteins 3 (AKAP3) and Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase 3 (PLOD3) gene mutations and prediction of 3D protein structure for ligand binding activity in the cases of non-obstructive azoospermic male. Materials and methods Clinically diagnosed cases of non-obstructive azoospermia (n=111) with age matched controls (n=42) were included in the present case-control study for genetics analysis and confirmation of diagnosis. The sample size was calculated using Epi info software version 6 with 90 power and 95% confidence interval. Genomic DNA was isolated from blood (2.0 ml) and a selected case was used for whole exome sequencing (WES) using Illumina Hiseq for identification of the genes. Bioinformatic tools were used for decode the amino acid sequence from biological database (www.ncbi.nlm.nih.gov/protein). 3D protein structure of AKAP3 and PLOD3 genes was predicted using I-TASSER server and binding energy was calculated by Ramachandran plot. Results Present study revealed the mutation of AKAP3 gene, showing frameshift mutation at rs67512580 (ACT → -CT) and loss of adenine in homozygous condition, where, leucine changed into serine. Similarly, PLOD3 gene shows missense mutation in heterozygous condition due to loss of guanine in the sequence AGG→A-G and it is responsible for the change in post-translational event of amino acid where arginine change into lysine. 3D structure shows 8 and 4 pockets binding site in AKAP3 and PLOD3 gene encoded proteins with MTX respectively, but only one site bound to the receptor with less binding energy representing efficient model of protein structure. Conclusion These genetic variations are responsible for alteration of translational events of amino acid sequences, leading to protein synthesis change following alteration in the predicted 3D structure and functions during spermiogenesis, which might be a causative “risk” factor for male infertility.

116-121

Authors: Khodakaram Jahanbin, Mehri Ghafourian, Mohammad Rashno

Objective Several studies have shown that leukemia inhibitory factor (LIF) is one of the most important cytokines participating in the process of embryo implantation and pregnancy, while, the role of this factor on vascular endothelial factor-A (VEGF-A), as one of the most important angiogenic factor, has not been fully investigated yet. The aim of this study was to evaluate the effect of LIF on gene expression of VEGF in the choriocarcinoma cells (JEG-3). Materials and methods In this experimental study, JEG-3 choriocarcinoma cells were treated with different concentrations of LIF (1, 10, and 50 ng) for 6, 12, 24, 48 and 72 hours. Expression of VEGF was analyzed by real-time PCR. Delta CTs were subjected to one-way analysis of variance (ANOVA) and a post hoc Tukey’s test by SPSS version 25.0 software for data analyzing. Results In the stimulated cells, different concentrations of LIF caused significant decrease of VEGF gene expression (P<0.05) at 12, 24 and 48 hours. In contrast, it was increased after 72 hours (P<0.001). Analysis of data after 6 hours also showed that level of VEGF gene expression was significantly decreased by increasing LIF concentration (P<0.001). Conclusion Expression level of VEGF gene was decreased in trophoblast cells (except after 72 hours) under the effect of different concentrations of LIF in a time-dependent manner. So, this study showed that further studies are needed to determine the effect of LIF on other angiogenic factors in trophoblast cells.

130-136

Authors: Gracia Lasheras, Gemma Mestre-Bach, Elisabet Clua, Ignacio Rodríguez, Borja Farré-Sender

Objective Cross-border reproductive care (CBRC) refers to the movement of patients to foreign countries for fertility treatment. Limited evidence indicates that this phenomenon is associated with a risk of psychological distress, but few studies on the psychological impact of CBRC are currently available. The aim of this study was to compare the anxiety and depression levels of a group of cross-border patients with a local Spanish patient group, both of which underwent in vitro fertilization (IVF) treatment. We also sought to explore the clinical, sociodemographic and personality profiles of the CBRC group and local women. Materials and methods This present cross-sectional study was conducted on 161 infertile females (71 CBRC patients and 90 local women) who were undergoing IVF treatment. The following questionnaires were used to collect data: Spielberger State Anxiety Inventory (STAI-S), the Beck Depression Inventory-II (BDI-II) and the Zuckerman-Kuhlman Personality Questionnaire (ZKPQ). Sociodemographic, clinical, reproductive and CBRC variables were also recorded. Results CBRC patients, specifically CBRC oocyte recipients, showed higher levels of anxiety compared to local women. However, no significant differences in depression scores were found between both groups. Finally, when analysing personality, the Activity scale scores of the ZKPQ were found to be higher in CBRC oocyte recipients, which indicated a greater tendency for general activity and higher energy levels. Conclusion CBRC oocyte recipient women may have greater vulnerability to anxiety than local women prior to infertility treatment. Screening and psychological support protocols for anxiety in this population should be considered.

143-149

Authors: Zahra Safaei, Shabnam Bakhshalizadeh, Mohammad Hossein Nasr-Esfahani, Azadeh Akbari Sene, Vahid Najafzadeh, Mansoureh Soleimani, Reza Shirazi

Objective Polycystic ovary syndrome (PCOS) is an endocrine disorder diagnosed by anovulation hyperandrogenism. Hyperandrogenism increases apoptosis, which will eventually disturb follicular growth in PCOS patients. Since mitochondria regulate apoptosis, they might be affected by high incidence of follicular atresia. This may cause infertility. Since vitamin D3 has been shown to improve the PCOS symptoms, the aim of study was to investigate the effects vitamin D3 on mtDNA copy number, mitochondrial biogenesis, and membrane integrity of granulosa cells in a PCOS-induced mouse model. Materials and methods In this experimental study, the PCOS mouse model was induced by dehydroepiandrosterone (DHEA). Granulosa cells after identification by follicle-stimulating hormone receptor (FSHR) were cultured in three groups: 1. granulosa cells treated with vitamin D3 (100 nM for 24 hours), 2. granulosa cells without any treatments, 3. Non-PCOS granulosa cells (control group). Mitochondrial biogenesis gene (TFAM) expression was compared between different groups using real-time PCR. mtDNA copy number was also investigated by qPCR. The mitochondrial structure was evaluated by transmission electron microscopy (TEM). Hormonal levels were measured by an enzymelinked immunosorbent assay (ELISA) kit. Results The numbers of pre-antral and antral follicles increased in PCOS group in comparison with the non-PCOS group. Mitochondrial biogenesis genes were downregulated in granulosa cells of PCOS mice when compared to the non-PCOS granulosa cells. However, treatment with vitamin D3 increased mtDNA expression levels of these genes compared to PCOS granulosa cells with no treatments. Most of the mitochondria in the PCOS group were spherical with almost no cristae. Our results showed that in the PCOS group treated with vitamin D3, the mtDNA copy number increased significantly in comparison to PCOS granulosa cells with no treatments. Conclusion According to this study, we can conclude, vitamin D3 improves mitochondrial biogenesis and membrane integrity, mtDNA copy number in granulosa cells of PCOS mice which might improve follicular development and subsequently oocyte quality

154-158

Authors: Maryam Hafezi, Zahra Chekini, Mohammad Reza Zamanian

Recurrent hydatidiform mole is defined as episodes of two molar pregnancies in a female. Often, complete moles only derive androgenic nuclear genome. We described two cases with repeated molar pregnancies attempted to prevent future episodes by performing intracytoplasmic sperm injection (ICSI) and preimplantation genetic diagnosis (PGD) to assess genetic disorders. The first patient had previously six complete molar pregnancies and advised to carry out ICSI with ovum donation to achieve a normal pregnancy. The second case had previously five molar pregnancies and no XY embryos from the ICSI/PGD process. We had to (at the insistence of the patient) transfer XX embryos in this patient which resulted in a complete hydatidiform mole (CHM). Hence, available data based on our patients and previous studies demonstrated that oocyte might play a critical role in the pathophysiology of recurrent hydatidiform mole, while it has not been often considered.