Journal of Shahid Sadoughi University of Medical Sciences
Year:2004 | Volume:11 | Issue:4|Papers Count:16

To improve the fertility potential of infertile women, gametes, embryo and ovarian tissue are preserved using cryopreservation. Ovarian cryopreservation has more advantages as compared to oocyte freezing. Ovarian tissue has a lot of follicles which can be used in sequential ovarian cycles. Ovarian preservation is useful for treatment of infertile patients undergoing chemotherapy and radiotherapy. The first reports about cryopreservation of ovarian tissue were published in 1950s and after that, human ovarian freezing was reported in 1995. There are three methods for freezing ovaries; slow freezing methods, vitrification and ultrarapid. In these techniques, the ice formation is inhibited for enhancing the viability of the cells after freezing. The main factors for freezing of tissue are cryoprotectants, velocity of cooling and freezing rate which are dependent on the kind of the cell and tissue, permeability of the membrane and the ratio of the surface and the volume of the cell. Transplantation and invitro development are used for development of cryopreserved ovarian follicles.  

Introduction: One of the most important steps in ART cycle is Embryo transfer followed by the sensitive stage of implantation, which depends on two main factors; embryo quality and receptivity of Endometrium. Another important factor in endometrial reception is existence of microorganisms in cervical canal which can involve normal flora of endometrium. They can have a harmful effect on implantation process by activating the immunity system upon entering the uterus. Materials & Methods: This descriptive and analytic study was done on 90 infertile patients aged between 20-40 years. Ovulation induction protocol was the same in all patients. In ovarian puncture, the first specimen was taken from the cervical canal before vaginal washing for microbial culture, while the second specimen was taken from cervical canal before any washing on transfer day after 48- 72hrs. The third specimen was from about 1.5-2 cm of transferred catheter that was cut and put in culture media. All of the specimens were cultured and evaluated for presence of gram positive and negative bacteria. Results: fertility rate in the two groups; the group whose culture specimen was sterile (31pt) and the one whose culture specimen was infected (49 patients) were not significant statistically. The group with a sterile catheter tip (29pts) was compared with the group with infected catheter tip (51pts). Fertility rate was 6.9% in the infected catheter group and 19.6%, in the sterile catheter group. Although fertility rate in sterile group was 2.8 times more than in the infected group, it was not statistically significant. Frequency of bacteria didn't effect the fertility rate. Fertility rate in the gram pos. group in comparison with the gram neg. group was 2.83 times, but not significant in the statistical analysis. Conclusion: In our study, infection of catheter tip with gram neg. bacteria had no statistically significant effect on fertility rate, but it still counts as a main factor for successful IVF/ ET results.    

Objective: Determination of incidence of cancellation of ART cycles, and factors influencing the cancellation. Data collection and materials/methods included 400 patients undergoing induction ovulation for ART. Incidence of cancellation of ART cycles was estimated. Results: Cancellation rate incidence was 10.5% that was higher in 35 year olds and older patients. Incidence of cancellation rate was higher in HMG user group and infertility due to tubal factor. The rate of cancellation was similar in patients with increasing drug dosage and those with fixed drug dosage. Rate of cancellation was also similar in both patients with more than two cycles and those with two cycles or less. Discussion: This study supports previous observations, correlations between age and ovarian response. Incidence of cancellation rate was higher in patients with tubal factor which supports the thesis correlating ovarian response and tubo-ovarian environment. Conclusion: Cancellation rate incidence was 10.5%. Poor responses of ovaries for developing follicles and absence of oocytes after retrieval were the most frequent factors responsible for cancellation rate.    

Introduction: Infertility has always been an unsolved puzzle. Fertility has had an important and critical role in human life and primitive cultures. As time passes and science & technology progress, infertility treatment has advanced beyond imagination, and one of the most noticeable progresses has been embryo culture invitro until the blastocyst stage. Materials & Methods: This study was done as a clinical trial in 30 infertile couples who had failed at least one cycle of ART without endocrine cause (case group) and 30 infertile couples were selected as control. In the case group, blastocyst transfer (BT), and in the control group, embryo transfer (ET) was done. Results: There was an insignificant difference between the 2 groups. ANOVA Variance analysis was used for statistical evaluation. Frequency distribution was done by x2 and Fisher exact test. Fertilization rate (FR) in HT group was 62% while it was 65% in the ET group. Pregnancy rate (PR) was 16.7% in BT group and 6.7% in ET group Conclusion: A treatment alternative for infertile women with previous failures is blastocyst transfer as clinical pregnancy can be increased upto 40-60% in ART cycles with blastocyst transfer.    

Introduction: Use of different methods for GnRH administration in ART cycles is a controversy.Microdose regimens in poor- responder patients are very useful and this has been determined by some studies, but it is not clear how this regimen affects patients with first (IVF/ICSI) cycle. Materials & Methods: In a prospective, randomized, clinical trial study at The Research and Clinical Center for Infertility and Madar Hospital, Yazd, we compared 140 patients who were candidates for IVF/ICSI for the first time. 70 patients received Microdose flare (A Group) and 70 patients were administered GnRH in long luteal phase (B group). Number of oocytes, follicles, embryos and the pregnancy outcome were compared in both groups. In microdose group, after administration of 21 contraceptive pills in the last cycle from day 3 , GnRH was given to patients ( 0.05 cc twice a day) and H MG from day 5 (3 vials a day). In long protocol, the patients received 0.5 cc Buserelin daily from day 21 subcutaneously and with the beginning of bleeding , the dosage was decreased to half. HMG was given three times a day. As the follow up of some patients was difficult and their pregnancy outcome was unknown, 14 patients of A group and 3 patients of B group were left out from the study. The data was analyzed by chi-square and T test. Results: Number of oocytes, embryos and outcome of pregnancy were not different the in two groups. However, the number of mature follicles in long method was higher but not significant statistically. Conclusion: The Microdose flare regimen thus offers no further advantage for patients with first cycle of IVF/ICSI.    

Introduction: There is a problem with unsuccessful treatment cycles in women who suffer from PCOS or poorly respond to clomoiphene citrate (CC). Considering the results of Mitwally MF et ai, 2001, we decided to evaluate the efficacy of aromatase inhibitors for induction of ovulation in such women. Material & Methods: According to the method, 20 women with PCOS who had not responded to CC participated in this study. A dose of 2.5 mg aromatase inhibitor letrozole was given orally on day 3-7 of cycles and sonography with estradiol level measurement were used to determine the ovulation status. Results: Only one of the twenty cases had one domain follicle (18 mm diameter) on day 12 of the cycle (Estradiol, 200 Pg/ml), IUI was then done without pregnancy outcome. In the remaining cases, repeated sonography was performed between day 9 & 15 of cycles and all cycles were cancelled due to absence of dominant follicle (> 14mm) E2 level in all cases were lower than 200 pg/ml and endometrial thickness was less than 8 mm. Conclusion: Our results did not confirm the favourable effects of letrozole used alone. In future, we are planning to use a combination regimen of letrozole plus CC or letrozole plus gonadotropin to evaluate the effect on similar patients.    

Introduction: Embryonic stem cells are pluripotent cells which are derived from early stage embryos and they have the ability to differentiate to several kinds of cells invitro. They are also used for genetic manipulation. Because of these properties, these cells are used in cell therapy and production of transgenic animals. The aim of this study was isolating and producing mouse embryonic stem cells from late blastocyst stage embryos for the first time in Iran for use in future investigations. Materials & Methods: 385 blastocyst stage embryos from pregnant NMRI mice were obtained and cultured 24-hours in DMEM medium supplemented with 10 % FBS. 2-4 days after hatching, inner cell masses formed colonies and they were recognized by their morphology and collected after tripsinization. Then, some of those cells were cultured on the primary mouse embryo fibroblast feeder layer and the others were cultured in the medium containing 0.1 mM b-Mercaptoethanol, 1000 u/ml LIF and 10% FBS. The embryonic stem cells were recognized by alkaline phosphatase histochemical technique. Results: Our results showed that highly pluripotent stem cells can be derived from blastocyst stage NMRI mouse embryos. Primary mouse embryo fibroblast feeder layer couldn't affect the growth and differentiation of embryonic stem cells. Conclusion: Overall, these embryonic stem cells can be used in future research such as producing of transgenic animals, gene therapy, gene targeting, etc.      

Introduction: Polycystic ovary syndrome (PCOS) is one of the most common endocrinologic diseases responsible for infertility. PCOS is characterized by AVB, infertility, hirsuitism & obesity. In this analytic study, a clinical trial was done about on PCOS women who attended the infertility center and hospital in 2001-2. Materials & Methods: We selected patients in our study and divided them to two groups; A (COH+timed intercourse) and B(COH+IUI). All patients received metformin 500-1500 mg/d for 2 months and then without cessation. In the third cycle, COH was done for them by clomphene 100 mg/day from 3rd to 8th day of menstural cycle and HMG 150 gr/day from 9th to 12th day of cycle. After controlling by TVS, when mature follicles reached 18 mm, HCG 1000 IU was injected, and then group A had normal Coitus while in group B, IUI was done. Results: Pregnancy rate (Positive BHCG test) after 2 weeks was 13% in group A and 16% in group B. This difference wasn't significant, but showed that IUI had better results than timed intercourse in PCO women. Conclusion: We therefore suggest (COH+ IUI) after 3 cycles of successful treatment with (COH+coitus) in infertile PCO.    

Introduction: GnRH-a+HMG combination is used for controlled ovarian hyperstimulation in IVF cycles. GnRH-a can be used in 2 ways;1) in follicular phase or 2) in luteal phase. There is discrepancy about its effect when used in the follicular phase. In this study, we determined the effects of microdose GnRH-a in Poor-responder patients. Materials & Methods: 61 poor-responders who were candidates for IVF or microinjection used low dose oral contraceptive pills for 21 days and then Bouserelin 40 µg subcutaneously 2 times /day from 3rd day of cycle and HMG 3Amp/day from 5th day of cycle. Datas were analyzed by SPSS chi –Square and Fisher Exact test. Results: Number of follicles, oocytes, embryo and pregnancy rate were determined, and compared with previous cycles. Difference was significant in all cases. (P < 0.05). Pregnancies occurred in 3 patients (5%). Conclusion: Results of this study showed that use of microdose GnRH-a +HMG for controlled ovarian hyperstimulation in IVF cycles can lead to formation of follicles, oocytes and embryoes in poor responders.    

Introduction: Heavy metals pollution of various resources and human exposure to these result in various disorders. It has been reported that chronic exposure to heavy metals has an adverse effect on reproductive and sperm functions. Aquatic ecosystem pollution with heavy metals due to industrial activities and release of city sewages in rivers is one of the major concerns of societies. Heavy metals accumulate in the beds of rivers, lakes and seas and then enter the food chain and are finally absorbed by humans. When they accumulate in reproductive tissues, they can cause deleterious effects on spermatogenesis and sperm morphology. Material & Methods: This study has been performed to study the ultra morphological changes in sperms exposed to 3 heavy metals. Sperms were incubated with different concentrations of cadmium, copper and mercury for 3 hours and were then analyzed by a scanning electromicroscope (SEM). Results: The results showed that mercury was the most toxic heavy metal studied here and caused tail detachment and head condensation in sperms. Cadmium and copper exerted extensive morphological effects on sperms (mainly enlarging sperm's head) and sperm decondensation. The results confirmed that part of heavy metals toxicity could be due to morphological changes and damaged sperms cannot swim freely and fertilize the ova. Conclusion: There was no difference in the sensitivity of the sperms of different varieties of fish towards the heavy metals.    

Introduction: Semen analysis including sperm count, progressive and non-progressive motility, and normal morphology of spermatozoa of ejaculates from infertile men are important. In general, assisted reproductive techniques (ART) are chosen according to the semen parameters. Therefore, it is crucial to evaluate semen parameters with application of one quick, standard, and inexpensive methodology. In some ART laboratories in Iran, the aforementioned analysis is done using traditional methodology such as wet preparation (microscopic slide technique) which can mislead the infertile couple. As a result, the infertile couple may not fully benefit from the treatment outcome. The objective of this prospective study was to compare the two techniques of Wet preparation and Makler - staining for analysis of semen parameters. Materials & Methods: The three parameters of count, morphology and motility of spermatozoa along with round cells of ejaculates from 50 men attending the infertility treatment program were evaluated. Following macroscopic evaluation, each seminal sample was evaluated with two different techniques of Wet and Makler - staining in a blind setting. For wet slide, semen parameters as well as round cells were studied. However, for Makler technique, Makler chamber was used for evaluation of percentage of motility and sperm count. In addition, Geimsa (WHO) staining was applied for evaluation of sperm morphology and round cells. The results showed that the volume, PH and viscosity of semen were within normal range of WHO. Results: The difference in the mean sperm count & percentage of motility was statistically insignificant. However, the sperm morpology was significantly different in the aformentioned groups (P=0.001). Also, the mean number of round cells in both groups was different (P=0.002). A total of one and nine seminal specimens were clear from round cells in Wet and Makler groups, respectively. The results showed that most variation was related to sperm morphology. For example, 5 seminal samples were abnormal in the wet method, while 32 ejaculates were abnormal in the Makler group. Conclusion: Sperm morphology' and round cells should be evaluated with more attention. Sperm morphology plays an important role in ART methodology and therefore should be counted and detected in full. It should be noted that Wet preparation technique may be used for evaluation of concentration and motility if Makler chamber is unavailable at ART laboratories.    

The most important aim of ART is to increase implantation to physiological levels, the same as a normal cycle. For this aim, two methods are recommended; transfer of best quality embryos and increase in endometrium receptivity. Materials & Methods: A Clinical trial study was performed on 160 cycles of ART( IVF/ICSI) at the Research and Clinical Center of Infertility and Madar Hospital of Yazd. With this Supposition that two step embryo transfer may increase the endometrial receptivity, two groups were compared. Both groups were similar in respect to male age, female age and duration and cause of infertility. For Induction -Ovulation using Long Protocol, Buserelin was injected subcutaneously, 0.5 cc daily from day 20 of menstrual cycle and the dose were decreased to 0.25cc daily at start of menstrual bleeding. HMG (225 IV/day) was injected from day 2 of cycle. The measurement of follicle diameter was done serially. After HCG injection (10000IU) and ovarian puncture, the luteal phase was supported by progesterone injection. Then, the patients were divided randomly in two groups. The first group of 80patients had experienced two step embryo transfer on day 3, and on day 5, two or three embryos was transferred. In the control group of 80 patients, embryo transfer was done routinely 48hours after ovarian puncture. The patients were followed up by measurement of 8HCG serum level on day 14 after embryo transfer. Results: Two patients of first group and 14 patients of control group were left out from the study because of no follow up. The data was analyzed by chi-square and t test. Finally, there was no significant difference in pregnancy rate in the two groups. (P-Value= 1.000). Therefore, Two Step embryo transfer dose not increase the pregnancy rate more than one step embryo transfer. Conclusion: There was no significant difference between the two methods and pregnancy rate, number of embryos & pregnancy rate, the female age and pregnancy rate. However, further studies are needed in this field.    

Introduction: The aim of this study was to determine the morphological and ultrastructural changes in mouse endometrium after ovarian hyperstimulation using HMG& HCG injection during implantation period. Materials & Methods: For this purpose, 10 N MRI mice, 6-10 weeks old were classified into two groups. One group was hyperstimulated and the other group considered as control. The samples which were obtained from 1/3 middle part of uterine horns were processed for light (H&E)and electron microscopic studies. Results: Our results show that hyperstimulation increases the height and density of surface and glandular epithelium. In epithelial cells of experimental group, lipid droplets were abundant. Conclusion: Thus, ovarian hyperstimulation using HMG and HCG caused some ultrastructural changes in the endometrial epithelium which could have an effect on the implantation of embryos and endometrial receptivity.    

Introduction: One of the most important aspects of every ART cyke is retrieval of good quality oocytes capable of good division that are accessed by selection of the best ovulation induction protocol on the basis of patients condition, age and cause of infertility. The aim of this study was evaluation of efficacy of low dose long acting GnRH-a (Decapeptyle) in prevention of premature LH surge and comparison of the outcome of ART with long protocol of short acting Gn-Rha (Buserekin). Materials & Methods: ln this prospective, randomized clinical trial performed in Yazd IVF center, a total of 60 patients with 61 cycles of ART were included in this study. Patients with endometriosis and those older than 40 years were excluded. Patients underwent COH-ET. 30 patients in group 1 received a single half dose of Decapeptyle (1.87mg) in mid-luteal phase and 31 patients in other group received Buserelin (0.5 mg) from previous mid - luteal phase which was reduced to 0.25mg at start of gonadotropin administration. The number of oocytes, embryo, fertilization, Cleavage, pregnancy and cancellation rate were compared in the 2 groups. Results: There was no case of cancellation due to premature LH surge in two groups, but one cycle in group 1 was cancelled due to follicular growth. There were no significant differences in fertilization, cancellation and pregnancy rate and the gonadotropin dosage was the same in both the two groups. Oocyte quality was better in group 2 (84.3% VS 77.2, P value <0.05), but cleavage rate (71.1% VS 81.9%,pvalue <0.01) and embryo development (85.6% VS 94.1%, P value<0.05) were lower in group 1 than long acting Gn-RH-a group. Conclusion: On the basis of findings, we concluded that low dose long acting Gn-RH-a is a useful method for pituitary suppression and facility of use and low cost of cycle are its benefits. On the other hand, long acting Gn-RH-a has no effect on oocyte quality, embryo development and luteal phase.