فهرست مطالب soheila arefi

We aimed to examine resveratrol effects on gene expression of Bcl-2, Bax and Bcl-2/Bax ratio in endometrial stromal cells derived from women with and without endometriosis.

Endometrial tissues were obtained from 40 endometriotic patients and 15 non-endometriotic controls undergoing laparoscopic surgery or hysterectomy in the gynecology ward of Rassoul Akram Hospital, Tehran, Iran from 2015 to 2017. After the enzymatic digestion, eutopic (EuESCs) and ectopic (EESCs) endometrial stromal cells from patients with endometriosis as well as endometrial stromal cells from non-endometriotic controls (CESCs) were treated with or without resveratrol (100 µM) and the levels of Bcl-2, Bax and Bcl-2/Bax gene expression ratio in the cells from all origins were examined at 6, 24 and 48 h post-treatment by real-time PCR.

Resveratrol treatment increased Bcl-2 expression in CESCs at 24 and 48 h and in EuESCs at 48 h (P<0.05), but had no significant effects on the expression of this gene in EESCs. On the other hand, resveratrol treatment increased Bax expression in EuESCs at 6 h and decreased its expression in EESCs at 48 h (P<0.05). Regarding the Bcl-2/Bax gene expression ratio, resveratrol treatment increased Bcl-2/Bax gene expression ratio in CESCs and EuESCs at 48 h (P<0.01). However, this treatment had no significant differential effect on Bcl-2 and Bcl-2/Bax gene expression ratio between CESCs and EuESCs at 48 h.

Resveratrol treatment significantly increased Bcl-2/Bax gene expression ratio in EuESCs and CESCs but had no significant effect in EESCs.

Family of colony-stimulating factors (CSF) have an essential role on early cross talk between embryo and uterine endometrium.

The aim of this study was to evaluate the effects of the single dose of Granulocyte-CSF (G-CSF) injection on clinical outcome of assisted reproductive technology cycle in patients with repeated implantation failures.

This randomized control trial study was performed on 52 infertile women who referred to the clinic with the history of more than three previous In vitro fertilization/Intracytoplasmic sperm injection-embryo transfer failures. All patients were stimulated with standard long protocol. All embryos were transferred on day five in blastocyst stage in both groups. The treated group received 300 μg (0.5 ml) recombinant human G-CSF subcutaneously which was injected 30 min before blastocyst embryo transfer.

There was not statistically significant differences in abortion rate in G-CSF and control group (p=0.09). G-CSF treated group showed higher clinical pregnancy rate in comparison with control group (56.2% vs. 40.0%) but it was not statistically significant (p=0.09). Although live birth rate in G-CSF group was higher than control group (53.1% vs. 35.0%) but there wasn’t statistically significant difference in the overall live birth rate between the two groups (p=0.10). G-CSF group had a twin pregnancies while in control group there was no twin pregnancy.

Our result demonstrates the possibility that pregnancy outcome is better in women with repeated unexplained In vitro fertilization failure who are treated with G-CSF.

Natural endometrium in Frozen-thawed Embryo Transfer (FET) may have some benefits upon implantation in patients with Repeated Implantation Failure (RIF). It might be due to possible differences between natural and stimulated endometrial growth factors and cytokins secretions.

The objective of this study was to compare the pregnancy rate of FET on modified natural cycle versus hormone replacement therapy (HRT) cycle endometrium in patients with RIF.

In this observational study the pregnancy rate of patients with RIF undergoing day 3 FET in natural cycle endometrium (group 1, n=56), were compared with another group of patients with RIF in whom frozen-thawed day 3 embryos were transferred on HRT cycle (group 2, n=52).

The pregnancy rate in group 1 was 41.07%, compared with the pregnancy rate of group 2; 36.5% (p=0.63). The abortion rate was not significantly different among the groups.

It can be concluded that FET in a modified natural cycle is comparable with HRT cycle in patients with RIF.

Various studies indicated that epithelial cells derived from the amniotic membranes of human term placenta have inhibitory effect on components of the innate or acquired immune system. On the other hand، during pregnancy one of the most inhibitory mechanisms of fetal survival maybe though to close contact between AECs and maternal myeloid derived (DCs) that leads to generate immunological tolerance avoid from tissue graft rejection by maternal immune defense. In the presence study we look out for generating tollerogenic DCs from human monocytes derived iDCs by co-culturing with human term amniotic isolated epithelial cells to demonstrate inhibitory effect of AECs on phenotyping changes and maturation of cultured DCs. This change included: expression of immature (CD14، CD1a) or mature (CD80، CD86، CD83) CD markers and MHC I، II by flow cytometer technique. The cultivation system improved by addition of LPS، IL4&GM-CSFgrowth factors and insert chamber contact manner during 7 days culture. Cytokine research was done by ELISA method (IL12،IL10). The surface expression of CD markers CD14، CD1a، CD80، CD86، CD83، HLA-DR and CD40) and secreted cytokines (IL10 and IL12) of cultural MDDCs during maturation of mature and immature DCs between two groups: test and controls (in the absent of AECs) no significant differences were observed. Any effects have been concluded from maturation and differentiation of MDDCs by co-culture with hAECs. Growing on future study by various changes in culture system our suggestion is implicate.

Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells (hAECs) remains controversial. The aim of the present study was to characterize immuno-phenotypic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of trypsin used for tissue digestion. Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four trypsin types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor (EGF). To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining. Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of trypsin used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies. Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC.

Human amniotic epithelial cells (hAECs) have been introduced as a valuable source of stem cells with therapeutic applications. Despite the extensive study and emerging evidence، cell surface marker signature of hAECs remained controversial. The aim of the present study was to establish an efficient and optimized isolation procedure of hAECs and their characterization in terms of proliferation and stem cell markers. Four human placentas were collected from healthy women undergoing elective caesarian delivery at term labor under sterile condition. Amniotic membranes were carefully isolated and subjected to trypsin digestion. Single cell suspensions were obtained and the epithelial origin was confirmed by assessment of cytokeratin expression. Expression of CD9، CD10، CD29، CD34، CD38، CD44، CD45، CD73، CD105، CD133، MHC-I، HLA-DR، HLA-G، SSEA-4، STRO-1 and OCT-4 was then evaluated by flow cytometry. The overall yield of hAEC ranged from 80-130×106 cells per placenta with more than 98% viability. Microscopic examination revealed that hAECs are large and refractive cells with great capacity to adhere to plastic surfaces. The cells substantially expressed cytokeratin implying their epithelial origin. Flow cytometry data revealed that hAECs are a heterogenic population consisting of immune phenotypically mixed cells. Nearly all purified cells expressed mesenchymal markers، CD9، CD10، CD29، and CD73. A large proportion of cells also expressed HLA-I، HLA-G، STRO-1، OCT-4 and SSEA-4. Isolated cells، however، failed to express CD34، CD38، CD44، CD45، CD105، CD133 and HLA-DR. The procedure presented here is a simple and cost-effective protocol for isolation of a large numbers of viable hAECs. Surface markers and such features as proliferation capacity of hAECs are among the characteristics that are strongly dependent upon isolation and cell culture conditions. Thus such variables should be taken in to consideration when stem cell properties of hAECs are to be interpreted.

Recurrent abortion (RA) may be a consequence of aberrant expression of immunological factors during pregnancy. Although the relative importance of immunological factors in human reproduction remains controversial, substantial evidence suggests that autoantibodies contribute to reproductive failure. Production of such antibodies is under the control of cytokines; and leptin, besides its role in reproductive success, has a profound effect on directing the cytokine profile toward Th1 (cellular) pattern. Therefore, the present study was performed to assess serum leptin levels in women with immunological recurrent abortion. In this prospective study, 250 women who attended Avicenna Infertility Clinic with RA were screened for known causes of abortion from July to December 2008 in Tehran, Iran. Eighty-one patients with normal karyotypes and hormonal profile with normal ovaries and uterus and no signs of infection were categorized as patients with immunological (IRA, n = 39) or unexplained (URA, n = 42) recurrent abortion based on presence or absence of autoantibodies. After blood sampling, levels of anti-nuclear antibody (ANA), anti-double stranded DNA antibody (anti-dsDNA), lupus anti-coagulant antibody (LACAb), anti-phospholipid antibody (APA), anti-cardiolipin antibody (ACA), anti-thyroglobulin antibody (TgAb), anti-thyroperoxidase antibody (TPOAb) and anti-thrombin III antibody (ATIIIAb) were measured by enzyme-linked immunosorbent assay (ELISA) or chemiluminescent enzyme immunoassay (CLEIA). In IRA group, 9 (23.1%), 24 (61.5%), 25(64.1%) and 1 (2.6%) women were above the normal cut-off point for ANA, TgAbs, TPOAbs and AT-III Abs, respectively. IRA patients had normal values of LACAbs, APA and ACA. With normal level of fasting blood sugar (FBS), IRA and URA groups had similar serum leptin levels (23.7 13.2 ng/ml vs. 22.7 12.5 ng/ml, respectively). Serum leptin concentrations showed a positive correlation with weight and BMI in both groups. This study suggests that serum leptin levels are higher in IRA and URA patients than normal women. The findings of this study suggest the need for a more comprehensive study and comparison of leptin levels in IRA and URA patients to women with no history of miscarriages.

The study aimed to evaluate the effectiveness of laparoscopic ovarian drilling (LOD) on ovulation and restoration of menstrual cycles and pregnancy. This cross sectional study was conducted on 81 consecutive Polycystic Ovarian Syndrome (PCO) patients whom were referred to Avesina Infertility clinic during 2003-2006. Exclusion criteria were tubal and male factor infertility, hyperprolactinemia, thyroid disease and diabetes mellitus. Patients with the age range of 20-35, body mass index of less than 35 kg/m2, failure to ovulate during at least three cycles of Clomiphene Citrate (CC) consumption and more than 12 months of ovulatory factor infertility entered the study. Metformin was used for at least 8 weeks in combination with CC. Forty four PCO women who had not ovulated after medical therapy with combination of CC and Metformin were selected for LOD. The data was analyzed with SPSS software using t- test and logistic regression analysis. P<0.05 was considered statistically significant. Patients aged 28.9±2.3 years with infertility duration of 4.8±2.3years and BMI of 28±3.3 kg/m2. Effectiveness of medical therapy significantly was related to BMI (P<0.001). After LOD, ovulation occurred in 14 cases (31.8%), restoration of menstrual cycle in 18 women (40.9%), and spontaneous pregnancy in 5 cases (11.3%). LOD is a good method for restoration of ovulatory cycles in anovulatory PCOS women who were resistant to the combination of CC and Metformin. LOD may avoid or reduce the need to gonadotropins for ovulation induction.

Neurocognitive dysfunction after cardiac surgery with cardiopulmonary bypass (CPB or “pump”) is a common complication, reported in up to 53% of patients. In many patients this morbidity is only mild and transient, but in 5% of cases, it is severe. In this prospective study from June 2002 to July 2004,186 cases underwent coronary artery bypass graft surgery (CABG) using CPB or off-pump CABG (OPCAB), and they were evaluated for neurocognitive complications by mini-mental status examination. The average age of the patients was 56±6.2 years, 62% were male and 38% were female. 121 operations were performed with CPB (on-pump) and 65 operations were done off-pump. Mini-mental status test was normal in 50% of off-pump CABGs and in 43% of on-pump CABGs, very mild disturbance was seen in 48% of off-pump CABGs and in 54% of on-pump CABGs (p value=0.192, NS), mild disturbance was seen in 2% of off-pump CABGs and in 3% of on-pump CABGs (p value 0.392, NS), and moderate or severe disturbance was not seen in either group. In our study, there was no significance difference in the frequency of postoperative neurocognitive complications between off-pump and on-pump CABG patients

Controlled ovarian hyperstimulation (COH) is an important factor in IVF/ICSI success rates. CRP is a serological marker for systemic inflammation and it increases following hormonal stimulation. Probable inflammation following ovulation induction and ovarian puncture, in regards to CRP changes, might affect the outcomes of IVF programs. The present study was conducted to determine the effects of ovulation induction and puncture of ovaries on serum CRP levels in IVF/ICSI candidates.

This observational, descriptive-analytical study was performed at Avicenna Infertility Clinic during 2006 – 2007. COH was performed on 70 infertile patients who were candidates for IVF/ICSI, using standard long GnRH agonist protocol. Peripheral blood was drawn four times during the treatment cycle on the first day of stimulation and the days of HCG injection, ovary puncture, and embryo transfer. Non-serosanguinous follicular fluid samples were taken at the time of ovum pick up from each individual too. CRP levels were measured in follicular fluid and serum using competitive ELISA methods.

Serum CRP levels increased along with the stimulation cycle, from the first day of the procedure to the day of HCG injection but significant increases were seen in 82.2% of the cases following ovary puncture (respectively, 3.97±1.00g/ml, 5.54±2.26g/ml and 6.61±4.16g/ml; p<0.001). There were no significant correlations between serum estradiol and CRP levels.

Controlled ovarian hyperstimulation and ovarian puncture potentiate systemic inflammation. Serum CRP levels increase during ovulation induction cycle and also after HCG injection, but not in all patients undergoing ovarian puncture. These inflammatory reactions may affect IVF/ICSI outcomes.

Polymorphisms of coagulation factor V gene are the most important suspected causes of thrombophilia in women with recurrent pregnancy losses (RPL). In this study the association between three polymorphisms of factor V (FV Leiden, FV A4070G & FV A5279G) and RPL are sought in Iranian women.

In this case-control study, 100 female patients with at least two recurrent abortions were selected as the cases, and 100 healthy women with a history of two successful deliveries as the controls. Peripheral blood samples were collected and DNA was extracted. PCR-RFLP method was used for genotyping the samples.

Regarding the prevalence of FV Leiden mutation in the cases and the controls, 13% and 4% respectively, the chances for recurrent pregnancy losses were more than 3.5 times higher in individuals with this polymorphism (OR: 3.586, 95% CI: 1.127–11.412). The frequencies of FV A4070G and FV A5279G were 14% and 37% in the case and 4% and 7% in the control groups, respectively and the chances for RPL were higher in cases with these two polymorphisms. The proportion of cases with two or three mutations in the gene in comparison with the controls, showed a significant correlation between FV Leiden and FV A4070G polymorphisms. Statistical analysis of the simultaneous effects of the three polymorphisms for RPL showed that evaluation of FV A4070G and FV A5279G could help assess the chances of the three mutations for RPL.

The three polymorphisms in coagulation V gene are accompanied with increased risks for RPL. Evaluation for the three polymorphisms is suggested in the work up of women with RPL.

Despite numerous developments in the field of assisted reproduction, the implantation rate remains low. Among the various reasons of implantation failure, endometrial regularity has an important role. Hysteroscopy is an accurate method for evaluating the endometrial characteristics, with the ability to treat uterine pathology.

The aim of the present study was to evaluate the findings on hysteroscopy and thereafter the result of subsequent IVF/ICSI in infertile women with the history of frequent unexplained and unsuccessful IVF/ICSI attempts.

In this observational study, the hysteroscopy findings and the outcomes of subsequent IVF/ICSI were evaluated in 89 infertile women admitted in Avicenna Infertility Clinic, with previous repeated (more than two) failed IVF/ICSI-ETs, including the patients with normal Hysterosalpinography (HSG) and excluding severe male factor infertility and also thrombophilia, genetic and immunologic problems. The data were analyzed with SPSS software and Fisher exact, chi-square, and MC-Nemar tests.

In 53 (59.5%) cases, hysteroscopy revealed abnormal intrauterine findings including adhesions 7 (13.7%), single polyp 11 (20.7%), endometrial polyposis 10 (18.8%), endometrial hyperplasia 10 (18.8%), uterine cavity hypoplasia 4 (7.8%) and myoma 5 (9.8%). These abnormalities were significantly higher in women with more than 8 years of infertility (chi-square=4.7, p-value=0.03). After hysteroscopy and subsequent IVF/ICSI-ET attempt using standard long protocol, pregnancy rate were significantly higher compared with the previous repeated IVF/ICSI attempts (35.8% versus 0%).

According to this study, we strongly suggest evaluation of endometrial integrity by hysteroscopy in patients with repeated IVF/ICSI-ETs failure, before entering any other fertilization procedures.

In spite of the great deal of research in assisted reproductive techniques, more than 80% of transferred embryos by IVF/ICSI methods fail to be implanted. The causes for repeated implantation failures (RIF) may be reduced endometrial receptivity or other various uterine pathologies, such as thin endometrium, altered expression of adhesive molecules or immuno-logical factors; whereas genetic abnormalities of male or female individuals, sperm defects, embryonic aneuploidy or zona hardening are other etiologies for implantation failures. Clinically, endometriosis, polycystic ovaries and hydrosalpinx may decrease implantation following embryo transfer due to dual disorders in the quality of embryos or endometrium. In this study, probable causes and methods of evaluation for RIF patients have been reviewed and the suggested methods for their treatment, including myomectomy, endometrial stimulation, immunotherapy, hysteroscopy, preimplantation genetic screening (PGS), assisted hatching, zygote intra-fallopian transfer (ZIFT), co-culture, blastocyst transfer, cytoplasmic transfer, tailoring stimulation proto-cols, intracytoplasmic sperm injection (ICSI) and salpingectomy for hydrosalpinges have been discussed.

This is a case of a modified Blalock-Taussig shunt, which was complicated by perigraft transudative, fibrinous fluid accumulation and recurrence after surgical intervention. Follow-up and expectant management of the patient was successful. Our experience regarding this complication is presented.