The distribution and possible role of ERK8 in mouse oocyte meiotic maturation and early embryo cleavage.

It is well known that extracellular signal-regulated kinase 8 (ERK8) plays pivotal roles in various mitotic events. But its physiological roles in oocyte meiotic maturation remain unclear. In this study, we found that although no specific ERK8 signal was detected in oocyte at the germinal vesicle stage, ERK8 began to migrate to the periphery of chromosomes shortly after germinal vesicle breakdown. At prometaphase I, metaphase I (MI), anaphase I, telophase I, and metaphase II (MII) stages, ERK8 was stably detected at the spindles. By taxol treatment, we clarified that the ERK8 signal was stained on the spindle fibers as well as microtubule asters in MI and MII oocytes. In fertilized eggs, the ERK8 signal was not observed in the two pronuclei stages. At prometaphase, metaphase, and anaphase of the first mitosis, ERK8 was detected on the mitotic spindle. ERK8 knock down by antibody microinjection and specific siRNA caused abnormal spindles, failed chromosome congression, and decreased first polar body extrusion. Taken together, our results suggest that ERK8 plays an important role in spindle organization during mouse oocyte meiotic maturation and early embryo cleavage.

[Study on the relationship between altered expression of annexin A4 and endometrial receptivity during the implantation window in infertile patients with endometriosis].

OBJECTIVE: To identify the differential expressed proteins, and to investigate the relationship between altered expression of annexin A4 during window of implantation [WOI (at day-6 after ovulatory day)] in infertile patients with endometriosis and endometrial receptivity. METHODS: Two-dimensional fluorescence differential in-gel electrophoresis (2D-DIGE) and matrix-assist laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) were used to detect protein expression in endometrial WOI in 10 infertile cases with endometriosis as endometriosis group and 10 infertile cases with tubal factors as control group. The semi-quantitative validation of annexin A4 in the eutopic endometrial tissue during WOI was analyzed by western blot. RESULTS: By comparing protein profiles, there were 7 meaningful differential proteins during WOI in infertile patients with endometriosis. One protein with an isoelectric point of 5.84 and relative molecular weight of 36 100 were down regulated 348% in samples of endometriosis group. It was identified as annexin A4 by mass spectrometry. By western blot, relative intensity of annexin A4 in endometriosis group was 7.2 ± 0.9, which was lower than 17.8 ± 2.6 in control group significantly (t = 7.654, P = 0.002). CONCLUSION: Lower expression of annexin A4 during WOI in infertile patients with endometriosis might be associated with the decrease of endometrial receptivity.

Abnormal expression of uncoupling protein-2 correlates with CYP11A1 expression in polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) may result from hypersensitivity to insulin, which is negatively regulated by uncoupling protein (UCP)-2. Because cholesterol side-chain cleavage enzyme (CYP11A1) is closely linked to PCOS, the expression of UCP-2 and CYP11A1 in ovarian tissues from PCOS patients was examined in the present study. Twelve PCOS patients with hyperandrogenaemia who underwent laparoscopic ovarian wedge resection and 12 age-matched control patients who underwent contralateral ovarian biopsy were enrolled in the study. UCP-2 expression in early stage (primordial, primary and secondary) and late stage (sinus and mature) follicles was examined using immunohistochemistry, whereas UCP-2 and CYP11A1 mRNA and protein levels in ovarian tissue were determined using quantitative reverse transcription-polymerase chain reaction and western blot analyses, respectively. UCP-2 expression increased significantly with follicular development in both control and PCOS tissue, with expression in early stage follicles from PCOS patients significantly greater than that in controls. In addition, both UCP-2 and CYP11A1mRNA and protein levels, mean fasting blood glucose concentrations and fasting serum insulin levels were significantly higher in PCOS patients compared with the control group. Finally, a significant correlation between UCP-2 and CYP11A1 expression was found in PCOS but not control patients. In conclusion, in PCOS patients, there was a correlation between UCP-2 and CYP11A1 expression, which was significantly higher than in the control group. These changes in UCP-2 and CYP11A1 expression may mediate follicle development in PCOS.

[Preliminary study on the value of ratio of serum luteinizing hormone/follicle-stimulating hormone in diagnosis of polycystic ovarian syndrome among women with polycystic ovary].

OBJECTIVE: To investigate the value of ratio of luteinizing hormone (LH) to follicle-stimulating hormone (FSH) in diagnosis of polycystic ovarian syndrome (PCOS) among women with polycystic ovary (PCO) and to compare the difference of the diagnostic criteria between the Rotterdam Consensus and the Committee for Reproductive and Endocrine in Japan Society of Obstetrics and Gynecology. METHODS: By means of transvaginal Doppler ultrasound, 195 women with PCO were diagnosed in Nanfang Hospital of Reproductive Medicine Center and compare difference of multiple clinical indexes according to Rotterdam consensus and Japan consensus respectively. In the mean time, the ratio of LH/FSH, the level of LH, testosterone (T) and recevier operating characteristic (ROC) curve were explored to on the value of diagnosis of PCOS. RESULTS: By Rotterdam consensus, 144 women were diagnosed with PCOS and 51 women were non-PCOS, while 111 were identified as PCOS and 84 were non-PCOS according to Japan consensus. LH/FSH in PCOS and non-PCOS were 1.59 ± 0.84 and 0.85 ± 0.47 respectively when based on Rotterdam consensus, and this ratio were 1.87 ± 0.76 in PCOS and 0.78 ± 0.39 in non-PCOS based on Japan consensus. When using LH/FSH to diagnosis PCOS by Rotterdam consensus and Japan consensus, areas under ROC curve are 0.786 and 0.942, respectively. CONCLUSIONS: The ratio of LH/FSH ≥ 1 provide the significant value in the diagnosis of PCOS. The criteria of the Committee for Reproductive and Endocrine in Japan Society of Obstetrics and Gynecology is more suitable for Chinese patients.

Demethylation of LHR in dehydroepiandrosterone-induced mouse model of polycystic ovary syndrome.

The cause of polycystic ovary syndrome (PCOS), a complex endocrine disorder, is unknown, but its familial aggregation implies underlying genetic influences. Hyperandrogenemia is regarded as a major endocrine character of the PCOS. In this study, we employed bisulfite sequencing and bisulfite restriction analysis to investigate the DNA methylation status of LHR, AR, FSHR and H19 in dehydroepiandrosterone (DHEA)-induced mouse PCOS model. The result showed that methylation of LHR was lost in ovary from induced PCOS mouse. However, AR, FSHR and H19 had similar methylation pattern in DHEA-treated group and control groups. These data provide evidence for close linkage between DNA demethylation of LHR and PCOS.

[Clinical outcome of patients with follicular development retardation by prolonged duration of gonadotropin administration for in vitro fertilization].

OBJECTIVE: To investigate prolonged use of gonadotropin (Gn) stimulation for the patients with follicular development retardation during controlled hyperstimulation (COH) for in vitro fertilization (IVF) cycles. METHODS: The inclusion criteria to categorize the good responder were that duration of Gn stimulation was < or = 15 days, total Gn dose used was < or = 3300 IU and total oocytes retrieved were > or = 4. The inclusion criteria for prolonged duration of Gn stimulation were that duration of stimulation was > or = 16 days. There were 69 oocyte retrieval cycles and 66 transfer cycles in prolonged stimulation group (group 1) and 483 oocyte retrieval cycles and 464 transfer cycles in good ovarian response group (group 2). The clinical characteristics and outcomes of two groups were analyzed. RESULTS: Clinical 5 pregnancy rate, implantation rate and delivery rate were 45.5% (30/66) versus 51.7% (240/464), 30.5% (385/1262) versus 28.0% (46/164) and 37.9% (25/66) versus 39.4% (183/464), respectively, in groups 1 and 2. Duration of Gn stimulation and dose were (20.8 +/- 4.2) and (10.3 +/- 1.8) days, (3090 +/- 1140) and (2302 +/- 862) IU in groups 1 and 2, respectively. There was no statistical difference in patient's age, basal follicular stimulating hormone (FSH), clinical pregnancy rate, implantation rate, early miscarriage rate, ovarian hyperstimulation syndrome (OHSS) rate, delivery rate between two groups (P > 0.05). There were more polycystic ovary (PCO) and (or) polycystic ovarian syndrome (PCOS) patients, more basal antral follicles, longer duration of Gn stimulation (range 16 - 33 days), higher Gn dose, lower serum peak estradiol (E(2)) level, fewer oocytes, fewer embryos transferred, in group 1 compared with group 2 (P < 0.01). CONCLUSIONS: These results demonstrate that prolonged Gn stimulation more than 16 days is a valuable alternative before cancellation of the IVF cycles for follicular development retardation during COH. Good clinical outcome can be achieved including pregnancy rate, implantation rate and delivery rate.

Expression of imprinted genes related to Beckwith-Wiedemann syndrome in human oocytes and preimplantation embryos.

OBJECTIVE: To investigate the expression of imprinted genes related to Beckwith-Wiedemann syndrome (BWS) in human oocytes and preimplantation embryos for understanding the relationship between assisted reproductive technology (ART) and BWS. METHODS: Using nested reverse transcription-PCR to analyze the expression of P57KIP2, LIT1, TSSC3 in human oocytes and preimplantation embryos. RESULTS: Transcripts of P57KIP2 were detected in human oocytes and at all stages of preimplantation embryos. LIT1 was expressed only in stages of 8-cell and blastocyst. Transcripts of TSSC3 could not be detected in human oocytes and preimplantation embryos. CONCLUSION: Transcripts of P57KIP2 and LIT1, imprinted genes related to BWS, were detected in human preimplantation development; ART might affect the epigenetics of imprinted genes in early embryogenesis.

[Effect of enucleation of hysteromyoma by laparoscopic surgery on protein oxidation and lipid hyperoxidation].

OBJECTIVE: To evaluate the effect of laparoscopic surgery on and lipid hyperoxidation in patients with hysteromyoma. METHODS: Forty patients with hysteromyoma were randomly divided into 2 equal groups: laparoscopy group and laparotomy group. The plasma advanced oxidation protein products (AOPP), malondialdehyde (MDA), antioxidant activity (AOA), and glutathione peroxidase (GPx) activity were measured before operation, just after operation (5 minutes after deflation) and 24 hours after operation. RESULTS: (1) In the laparoscopy group, the levels of AOPP and MDA were (50.20 +/- 9.23) micromol/L and (1.85 +/- 0.19) micromol/L before operation, increased significantly just after operation [(68.75 +/- 12.69) micromol/L and (2.52 +/- 0.55) micromol/L respectively, both P < 0.01], and recovered to the normal level 24 hour postoperatively [(49.70 +/- 9.92) micromol/L and (2.05 +/- 0.68) micromol/L respectively, both P > 0.05]. The levels of GPx and AOA decreased significantly just after operation [(0.29 +/- 0.09) U/ml vs. (0.62 +/- 0.27) U/mL and (0.90 +/- 0.24) mmol/L vs. (1.41 +/- 0.39) mmol/L respectively, both P < 0.01], and the GPx level recovered 24 hours after operation [(0.52 +/- 0.06) U/mL, P > 0.05], however, the AOA level was still lower [(1.00 +/- 0.31) mmol/L, P < 0.01]. In the laparotomy group, the levels of plasma AOPP and MDA level slightly increased just after operation in comparison with those before operation [(53.39 +/- 9.86) micromol/L vs. (52.30 +/- 7.10) micromol/L and (2.09 +/- 0.51) micromol/L vs. (1.83 +/- 0.64) micromol/L respectively, both P > 0.05] and continued to increase 24 hours after operation [(63.40 +/- 15.5) micromol/L, P < 0.05, and (2.42 +/- 0.44) micromol/L, P < 0.01]; the GPx and AOA levels decreased a little just after operation [(0.51 +/- 0.17) U/mL vs. (0.57 +/- 0.21) U/mL and (1.20 +/- 0.46) mmol/L vs. (1.33 +/- 0.37) mmol/L, both P > 0.05] and continued to decrease 24 hours after operation [(0.35 +/- 0.19) U/mL and (0.92 +/- 0.22) mmol/L respectively, both P < 0.01]. Compared with those of the laparotomy group, the plasma AOPP and MDA levels of the laparoscopy group were both significantly lower (P < 0.01 and P < 0.05), and the GPx level was significantly higher (P < 0.01) 24 hours after operation, however, the AOA level was not significantly different (P > 0.05). CONCLUSION: Laparoscopic surgery is better than laparotomy. Protein oxidation and lipid hyperoxidation occur during the laparoscopic surgery, however, disappeared after operation. Free radicals are generated by the end of laparoscopic procedure, possibly as a result of an ischemia-reperfusion phenomenon induced by the inflation and deflation of the pneumoperitoneum. AOPP and MDA are induced during laparoscopic procedure and then return to the normal levels finally.

[Predictive value of endometrial ultrasonography and age for the outcome of in vitro fertilization-embryo transfer].

OBJECTIVE: To assess the predictive value of endometrial ultrasonography and age for the outcome of in vitro fertilization-embryo transfer (IVF-ET). METHODS: A total of 305 women with tubal occlusion were studied. Superovulation was induced and transvaginal ultrasonography was employed for monitoring the endometrial thickness and endometrial patterns on the day of human chorionic gonadotrophin (hCG) administration. Pregnancy was diagnosed by ultrasonography of the intrauterine gestational sac. RESULTS: No difference was noted in the endometrial thickness or endometrial patterns between pregnant and nonpregnant groups, but the least endometrial thickness essential for pregnancy was 7 mm. The pregnant rate of women aged 20-34 years was higher than that of women aged 35 years or above. CONCLUSIONS: Neither endometrial thickness nor endometrial patterns detected by transvaginal ultrasonography on the day of hCG administration allows a reliable prediction of pregnancy. The least endometrial thickness essential for pregnancy is 7 mm and the pregnant rate of women aged 20-34 years was higher than those aged 35 years and above.

[Expression of cathepsins B and L in early gestational decidua and chorionic villi].

OBJECTIVE: To study the expression of cathepsins B and L in first-trimester gestational decidua and chorionic villi. METHODS: The decidua and chorionic villi in the first trimester of gestation were obtained from 30 women undergoing induced abortion, 25 with spontaneous abortion, 10 with normal endometrium in secretory phase, and 15 with bydatidiform mole in whom the expression of cathepsins B and L was determined by immunohistochemistry. RESULTS: Positive staining for cathepsins B and L were mainly detected in the trophoblasts and decidual cells. The positive expression rate of cathepsin B in the normal endometrium of secretory phase, decidua in the first trimester of gestation and spontaneous abortion were 10% (1/10), 83.3% (25/30), and 32.0% (8/25), respectively, and the rate of cathepsin L expression in the endometrium in secretory phase, decidua of induced abortion and spontaneous abortion were 0.0%, 63.3% (19/30), and 32.0% (8/25) respectively, showing significant difference in the expression rates of cathepsins B and L between the 3 groups (P<0.05). Strong positive expression of cathepsin B in chorionic villi of induced abortion, spontaneous abortion and hydatidiform mole were detected at the rates of 10.0% (3/30), 0.0%, and 66.7% (10/15), respectively, and cathepsin L at the rates of 26.7% (8/30), 4.0% (1/25), and 80.0% (12/15), respectively, with significant difference in strong cathepsins B and L expressions between the 3 groups (P<0.05). CONCLUSION: The expression of cathepsins B and L differs in normal and abnormal deciduas and chorionic villi of early pregnancy, suggesting that the cathepsins may play important roles in the process of implantation.

[Expression of HLA-G protein in trophoblast cells].

OBJECTIVE: To investigate the expression of human leucocyte antigen protein G (HLA-G) in different trophoblast cells and different stages of pregnancy. METHODS: The expression of HLA-G protein in normal placenta and trophoblasts of different trimesters was detected using immunohistochemical method (SP). RESULTS: HLA-G protein expression exhibited spatio-temporal changes, which located in the extravillous trophoblast (EVT) and was higher in the placenta of the first and second trimesters while lower in the third trimester (P<0.01). CONCLUSION: HLA-G protein expression in different stages of pregnancy and different trophoblasts may be related to the controlled invasion of the trophoblast.

[Significance of pelvic lymph node human papillomavirus DNA detection in cervical cancer].

OBJECTIVE: To assess the significance of human papillomavirus (HPV) DNA detection in the pelvic lymph nodes in cervical cancer. METHODS: HPV L1 gene fragment was amplified by HPV-specific PCR with general consensus primers from cervical cancer tissues. The types of HPV were identified by sequencing of the PCR product. The relationship between HPV positivity and pathological findings in metastatic pelvic lymph nodes of cervical cancer was analyzed. RESULTS: The positivity rate of HPV DNA was 40% in the pelvic lymph nodes in 40 cases of cervical cancer. HPV DNA was detected in the pelvic lymphatic nodes of all the 10 cases with pathologically confirmed lymph node metastasis of cervical cancer. HPV18 was detected in both of cervical and pelvic lymph nodes in some cases. CONCLUSIONS: HPV DNA can be detected in the pelvic lymph nodes before pathological identification of lymph node metastasis and indicates early pelvic lymph node metastasis of cervical cancer. The presence of HPV18 often indicates high likeliness of lymph node metastasis and suggests poor prognosis of the patients.

[Sequence polymorphism of human papillomavirus type 16 E7 in cervical cancer].

OBJECTIVE: To explore the genetic polymorphism of E7 open-reading frame of human papillomavirus (HPV) type 16 in cervical cancer. METHODS: The types of HPV was identified by sequence analysis of the PCR product of HPV in cervical cancer tissues. HPV 16 gene fragment in the cervical cancer tissue was amplified by HPV-specific PCR with general consensus primers. RESULTS: The positivity rate of high-risk HPV types in 50 cases of cervical cancers was 78%. Mixed infection of HPV16 and HPV18 was found in 18 cases, and the infection with HPV16 alone occurred in 15 cases. HPV16 E7 was amplified from 25 out of the 34 cases positive for HPV16. A T-to- C change occurred in the 647th nucleotide in the viral nucleotide sequence, causing conversion of the Asn codon of E7 gene into a Ser codon. CONCLUSIONS: The most frequently observed substitution in HPV16 E7 open reading frame occurs in the discrete regions of 647-846, and some substitutions result in the same-sense mutation. The hot-spot mutation of HPV16 E7 in cervical cancers in Guangdong Province occurs at the nucleotide 647 and 846 in the DNA sequence.

[Expression of imprinted gene Grb10 in human oocytes and preimplantation embryos].

OBJECTIVE: To investigate the mRNA expression of imprinted gene Grb10, a candidate gene for Silver-Russell syndrome (SRS), in human oocytes and preimplantation embryos for understanding the role of Grb10 in embryogenesis and assisted reproductive technology (ART). METHODS: Nested reverse transcriptase (RT)-PCR was used to analyze Grb10 gene expression in human oocytes and preimplantation embryos. RESULTS: The transcripts of Grb10 were detected in human oocytes and preimplantation embryos at all stages except for the 2-cell stage. CONCLUSION: Grb10 gene is expressed in preimplantation embryos with close association with embryogenesis.

[Association of the novel hydatidiform mole-related gene F10 with the invasiveness of trophoblastic tumor].

OBJECTIVE: To study the expressions of the novel gene F10 associated with hydatidiform mole in different trophoblastic tumors and explore the relation of F10 expression with the invasiveness of malignant trophoblastic tumor. METHODS: In situ hybridization was used to study the expression of F10 in 12 cases of hydatidiform mole, 6 cases of invasive mole, and 8 cases of choriocarcinoma. RESULTS: F10 mRNA was positive in all cases of hydatidiform mole, invasive mole, and choriocarcinoma, and the expression intensity significantly increased in the order of hydatidiform mole, invasive mole and choriocarcinoma (P<0.001). CONCLUSION: The expression of F10 gene may relate to the occurrence and invasiveness of trophoblastic tumor, with possible involvement in the invasion or malignant changes of trophoblastic cells.

[Detection of human papillomavirus DNA in cervical cancer tissue by PCR and direct sequencing].

OBJECTIVE: To explore an effective method for detecting human papillomavirus (HPV) DNA in cervical cancer tissue. METHODS: HPV L1 gene fragment in cervical cancer tissue was amplified by HPV-specific PCR with consensus primers, and typing of the HPV strains was performed on the basis of sequence analysis of the PCR product. RESULTS: The positivity rates of HPV DNA was 78% in the 50 cases of cervical cancer, and mixed infection with HPV16 and HPV18 strains was the most common, which accounted for 48% on the total infections. Infection with HPV58 was detected in one case. The sequencing results showed no difference in L1 sequence between the detected samples and the standard German HPV58 strain. CONCLUSION: PCR and direct sequencing approach is effective for detecting and typing of HPV DNA in cervical cancer tissue, through which rare HPV strain or mutants of known HPV strains may not escape detection.

[Expression of E-cadherin in human embryo].

OBJECTIVE: To investigate the role of calcium dependent adhesion molecules (cadherin) during the implantation and development of human embryo by studying the expression of E-cadherin in human embryo and blastocysts. METHODS: Expression of E-cadherin in preimplantation embryos and blastocysts was detected by indirect immunofluorescence assay and quantitated by laser scanning confocal microscopy. RESULTS: Expression of E-cadherin was found in all the preimplantation embryos and blastocysts the present authors studied. The expression was higher in blastocysts than that in preimplantation embryos. CONCLUSION: The above results suggested that human embryos and blastocysts could express E-cadherin and the expression increased during their development. These may be of significance to the adhesion and implantation of the human embryo and blastocysts.

[Mitochondria transfer from self-granular cells to improve embryos' quality].

OBJECTIVE: To improve embryos' quality and pregnancy rate with the method of self-mitochondria transfer. METHODS: All the 18 cases of women with repeated in vitro fertilization-embryo transfer (IVF-ET)-treatment failure or older than 37 years were treated with the long gonadotropin releasing hormone-agonist (GnRH-a) regimens. The oocytes were divided into two groups: intracytoplasmic sperm injection (ICSI) group and mitochondria transfer group if the number of the individual patients' oocytes was more than four. In mitochondria transfer group, mitochondria were prepared by different centrifugation after the granulosa cells were trimmed from the oocytes or collected from follicular fluid and homogenated. Mitochondria and immobilized sperm were injected into oocytes by micromanipulation. The fertilization rate and embryos' quality were compared. RESULTS: The fertilization rate was 74.4% in mitochondria transfer group, and 76.8% in ICSI group, with no statistical difference (P > 0.05). The good quality embryo rate in mitochondria transfer group was 59.4%, and it was 34.9% in ICSI group. The difference was significant (P < 0.05). There were 7 clinical pregnancies in the 18 cases. CONCLUSION: Self-mitochondria transfer can improve embryos' quality and pregnancy rate without influence on fertilization rate.

[Brain-derived neurotrophin factor inhibits steroid biosynthesis by human granulosa-lutein cells].

OBJECTIVE: To study the effect of brain-derived neurotrophin factor (BDNF) on the synthesis of estradiol and progesterone in human granulose-lutein cells (HGLCs) and the expression of steroidogenic acute regulatory factor (STAR) mRNA. METHODS: HGLCs were obtained from infertile women undergoing ovulation induction for fertilization-embryo transfer (IVF-ET) for male or tubal factors. HGLCs were cultured in serum-free media 199 for 24 h and treated by BDNF at 25, 50 and 100 ng/ml. Radio immunoassay (RIA) was used to examine the concentration of estradiol and progesterone, and reverse transcriptional PCR (RT-PCR) employed to detect the expression of STAR mRNA after treatment with BDNF at the concentrations of 25, 50 and 100 ng/ml. RESULTS: BDNF significantly inhibited the production of progesterone (P(4)) in the culture media of HGLCs in a dose-dependent manner. BDNF did not change the level of 17beta-estradiol (E(2)), but decreased the expression of STAR mRNA dose-dependently. CONCLUSIONS: BDNF can inhibit the synthesis of P(4) in HGLCs and regulate ovarian steroidogenesis. BDNF may inhibit HGLCs from producing P(4) by decreasing the transcription level of STAR gene in human ovary, and plays an important role in luteal regression.

[Normal pregnancy in a woman having had a child with trisomy 21 syndrome before: application of preimplantation genetic diagnosis].

OBJECTIVE: To study the value of preimplantation genetic diagnosis (PGD) before pregnancy for couples who are exposed to high risk of trisomy 21 syndrome in their progenies to obtain birth of healthy child by in vitro fertilization procedures. METHODS: Conventional hormone replacement treatment with intracytoplasmic sperm injection (ISCI) was administered for ovum fertilization. One or two cells (blastomeres) were aspirated from the preimplantation embryo that had grown to 6 to 10 cells (day 3 post fertilization) for PGD with fluorescence in situ hybridization (FISH), and 3 embryos with normal chromosome were transferred into the uterus. RESULTS: PGD was carried out in a total of 8 embryos with valid diagnoses in 7, of which 6 were identified as normal embryos and 1 had trisomy 21 syndrome. Pregnancy was obtained that resulted in the birth of a healthy baby. CONCLUSION: Detection of chromosomal abnormalities such as trisomy 21 syndrome is feasible for PGD with FISH.