Use of autologous buccal mucosa for vaginoplasty: a study of eight cases.

BACKGROUND: Vaginal agenesis is a rare condition and treatment methods are varied. The difference between most of the surgical techniques is the graft material used. The purpose of this study was to describe the procedure and outcome of creating a neovaginal pouch lined with autologous buccal mucosa. METHODS AND RESULTS: Between August 2000 and February 2002, eight patients with Mayer-Rokitansky-Kuster-Hauser syndrome were admitted to our hospital. All of the patients successfully underwent neovaginoplasty with autologous buccal mucosa as graft material. The buccal mucosal wound completely healed 2 weeks after the operation and the neovaginal length and calibre were well formed. Follow-up ranged from 0.5 to 1.5 years. One patient encountered post-operative vaginal bleeding and another patient suffered from urinary bladder injury. CONCLUSION: This is the first reported procedure of vaginoplasty with autologous buccal mucosa as graft material. Our method is ideal in its simplicity, provides good cosmetic results, and improves the vaginal length of the patient.

Zona-free oocyte fertilized with intracytoplasmic sperm injection and underwent further division: case report and literature review.

The zona pellucida (ZP) plays a protective role during fertilization and early embryonic development. It is related to sperm binding, the acrosome reaction, prevention of polyspermic fertilization, and holding blastomeres together before the morular stage. Zona-free oocytes are accidentally encountered. If these oocytes are healthy, they can be fertilized normally by intracytoplasmic sperm injection (ICSI). We reported on a couple with male infertility undergoing oocyte retrieval after ovarian hyperstimulation. Before the ICSI procedure, cumulus cells surrounding the oocytes were removed, which resulted in one oocyte escaping from its ZP. The zona-free oocyte was fertilized normally with ICSI and developed to the 8-cell stage. We observed that the zona-free zygote had the ability to further divide, despite its loose contact. The zona-free embryo was transferred with other zona-intact embryos, but the implantation failed. We conclude that zona-free oocytes can be rescued, fertilized with ICSI, and cultured for further transfer or cryopreservation.

Relation of the estrogen receptor alpha gene microsatellite polymorphism to bone mineral density and the susceptibility to osteoporosis in postmenopausal Chinese women in Taiwan.

OBJECTIVE: Osteoporosis is a common disorder with a strong genetic component. Our aim was to investigate the correlation of the estrogen receptor alpha gene microsatellite polymorphism (TA dinucleotide repeat polymorphism 5' upstream of exon 1) with bone mineral density and their relationship to osteoporosis. METHODS: We determined the estrogen receptor alpha gene microsatellite polymorphism using polymerase chain reaction-based microsatellite analysis in postmenopausal Chinese women in Taiwan. Bone mineral density of the lumbar spine and proximal femur were measured using dual-energy X-ray absorptiometry. RESULTS: The ERalpha genotype was classified into '12' through '27' according to the number of TA dinucleotide repeats they contained, as a 'signpost'. After adjustment for potential confounding factors including age, height, and weight, subjects with genotype 18+ (n=4) had lower bone mineral density values and a 54.5 times greater risk for osteoporosis when compared with subjects with genotype 18- (n=170) at the lumbar spine. This should be interpreted with caution because of the small number of subjects with the unfavorable genotype 18+. According to mean number of TA dinucleotide repeats, women with a high number of repeats (TA > or =20) (n=38) had the lowest bone mineral density and a 6.1 times greater risk for osteoporosis than women with a low number of repeats (TA < or =15) (n=61) at the femoral neck, after adjustment for potential confounding factors such as age, height, and weight. CONCLUSION: The present study suggests that the estrogen receptor alpha gene microsatellite polymorphism may be a candidate genetic marker for risk of osteoporosis in postmenopausal Chinese women in Taiwan.

Polymorphisms for interleukin-1 beta (IL-1 beta)-511 promoter, IL-1 beta exon 5, and IL-1 receptor antagonist: nonassociation with endometriosis.

PURPOSE: We aimed to investigate if interleukin-1 beta (IL-1 beta) and IL-1 receptor antagonist (IL-1Ra) gene polymorphism could be used as markers of susceptibility in endometriosis. MATERIALS AND METHODS: Women were divided into two groups: 1) endometriosis (n = 120); 2) nonendometriosis groups (n = 103). Polymorphisms for IL-1 beta-511 promoter, IL-1 beta exon 5, and IL-1Ra were detected by polymerase chain reaction. Genotypes and allelic frequencies for these polymorphisms in both groups were compared. RESULTS: Proportions of different IL-1 and IL-1Ra polymorphisms in both groups were nonsignificantly different. Proportions of C homozygote/heterozygote/T homozygote for IL-1 beta-511 promoter in both groups were 1) 21.6/59.1/19.1% and 2) 26.2/50.5/23.3%. Proportions of E1 homozygote/heterozygote/E2 homozygote for IL-1 beta exon 5 in both groups were 1) 91.6/5/3.3% and 2) 95.15/4.85/0%. Allele I/II/IV/V for IL-1Ra in both groups were 1) 92.5/5.4/1.6/0.4% and 2) 95.1/3.9/1/0%. CONCLUSIONS: Association of endometriosis with IL-1 beta-511 promoter, IL-1 beta exon 5, and IL-1 receptor antagonist gene polymorphisms doesn't exist. These polymorphisms are not useful markers for prediction of endometriosis susceptibility.

Kinetics of acetyl coenzyme A: arylamine N-acetyltransferase from human cumulus cells.

PURPOSE: N-acetyltransferase (NAT) activity is involved in the detoxification of exogenous amines. We aimed to evaluate the kinetics of acetyl coenzyme A (AcCoA): arylamine NAT for human cumulus cells. METHODS: Thirty infertile women who were undergoing controlled ovarian hyperstimulation (COH) and oocyte retrievals were recruited. Human cumulus cells were obtained during oocyte retrievals. Using 2-aminofluorene (2-AF) and p-aminobenzoic acid (PABA) as substrates, NAT activity and Michaelis-Menten kinetics constants of all samples were determined by using high-pressure liquid chromatography. RESULTS: There were 6 rapid, 10 intermediate, and 14 slow acetylators. 2-AF-NAT and PABA-NAT activities were 0.97 +/- 0.74 and 0.89 +/- 0.77 nmol/min/mg protein, respectively. Km/Vmax of rapid and slow acetylators for 2-AF were (161 +/- 55)/(15.6 +/- 2.9) and (27.8 +/- 11.4)/(2.6 +/- 0.9), respectively. Km/Vmax of rapid and slow acetylators for PABA were (104 +/- 36)/(13.2 +/- 2.8) versus (20.0 +/- 10)/(2.0 +/- 0.7), respectively. Compared to slow acetylators, the rapid acetylators exhibited higher Km/Vmax values for 2-AF (5.8-/6-fold) and PABA (6-/6.6-fold), respectively. CONCLUSION: Human cumulus could acetylate arylamine carcinogen (2-AF) and noncarcinogen drug (PABA). Higher percentage of rapid acetylators established in the cumulus during COH. It provides a model for monitoring the effects of pollution or carcinogenesis upon the oocyte during COH and oocyte retrievals.

Relation of polymorphism in the promotor region for the human osteocalcin gene to bone mineral density and occurrence of osteoporosis in postmenopausal Chinese women in Taiwan.

Osteoporosis is a common disorder with a strong genetic component. Our aim was to evaluate the correlation of the HindIII osteocalcin gene polymorphism to bone mineral density (BMD) and their relationship to osteoporosis. We determined the HindIII osteocalcin gene polymorphism using polymerase chain reaction (PCR)-based restriction analysis in postmenopausal Chinese women in Taiwan. The osteocalcin gene polymorphism was detected by the restriction enzyme HindIII, where the H allele indicated the absence of the cuttable site and the h allele indicated its presence. We then related the genotypes to BMD and occurrence of osteoporosis in these women. The allelic frequencies for postmenopausal Chinese women in Taiwan were 64% for h and 36% for H in HindIII restriction fragment length polymorphisms. The prevalence of each genotype in the study population was 37.7% hh, 52.6% Hh, and 9.7% HH. The subjects with genotype hh had the greatest BMD at the lumbar spine and the femoral neck, and those with HH had the smallest BMD at the femoral neck, but these differences did not reach statistical significance. The HindIII osteocalcin genotype showed a significant effect on the prevalence of osteoporosis in the subjects at the femoral neck, that is, women with genotype HH had a 6.4 times greater risk for osteoporosis (P < 0.05), and those with genotype Hh had a 1.2 times greater risk than women with genotype hh. In conclusion, the HindIII osteocalcin gene polymorphism is associated with reduced BMD and predisposes women to osteoporosis at the femoral neck.

Doppler study of the fetal middle cerebral artery at three locations: preliminary report.

BACKGROUND: Middle cerebral artery (MCA) detection is useful in monitoring fetal well-being. Knowledge of Doppler flow velocity of the fetal MCA may assist in prenatal diagnosis and management of complicated pregnancies. The aim of this study was to compare the pulsatility index (PI) and resistance index (RI) of the MCA at different locations throughout pregnancy. METHODS: Uncomplicated singleton pregnancies accepted Doppler surveys of the bilateral MCA. PI and RI values of the proximal, middle, and distal 1/3 of the MCA were measured. The gestation periods for Doppler surveys were (1) 15 to 19 weeks; (2) 20 to 24 weeks; (3) 25 to 29 weeks; (4) 30 to 34 weeks; and (5) 35 to 40 weeks. The MCA flows at different locations and at different gestational aged were compared. RESULTS: There were 21 patients included. Average PI/RI values of the proximal, middle and distal MCA were 1.62/0.80, 1.69/0.81, and 1.71/0.83, which were non-significantly different. The PI/RI values of MCA in each gestational phase were also non-significantly different: (1) 1.70/0.84; (2) 1.72/0.82; (3) 1.68/0.83; (4) 1.65/0.81; (5) 1.62/0.77. CONCLUSIONS: The PI and RI values of the proximal MCA were non-significantly lower than those of middle and distal MCA. Middle MCA could represent three locations of MCA. A trend of lower PI/RI values with advancing gestation was noted.

Antral follicle counting in predicting the retrieved oocyte number after ovarian hyperstimulation.

PURPOSE: To evaluate the antral follicle (AF) counting in predicting the outcome after controlled ovarian hyperstimulation (COH) and IVF-ET. METHODS: Infertile women who accepted the COH and IVF-ET were included prospectively. Day

Prognostic value of nm23 expression in stage IB1 cervical carcinoma.

BACKGROUND: The purpose of this retrospective study was to evaluate the patterns of nm23 expression in stage IB1 squamous cell carcinoma of the uterine cervix, to compare nm23 expression with clinicopathological findings and to assess its prognostic value. METHODS: Twenty-seven patients with stage IB1 squamous cell carcinoma of the uterine cervix underwent abdominal radical hysterectomy and pelvic lymph node dissection. Expression of nm23 was studied immunohistochemically, followed by amplification and direct sequencing of exons 4 and 5 of the nm23 gene. RESULTS: Overexpression of nm23 was detected in 18.5% of the tumors and low expression was seen in 33.3%, while negative expression was found in 48.1% of the tumors. Deep cervical stromal invasion (> or =1/2) was found to be associated with the increased risk of lymph node metastases (odds ratio = 17.5). A significantly lower percentage of patients survived when nm23 overexpression was observed (p = 0.0063). Univariate analysis revealed that tumor size (2-3.9 cm), lymph node metastasis, tumor invasion into parametria, tumor invasion into blood/lymph vessel, squamous cell carcinoma (> or =2 ng/ml) and nm23 overexpression had a significantly lower recurrence-free survival rate of the patients. None of the above factors was significant according to multivariate analysis. There were no mutations in exons 4 and 5 of the nm23 gene in stage IB1 squamous cell carcinoma of the uterine cervix. CONCLUSIONS: This study suggests that expression of nm23 may be indicative of an unfavorable prognosis in patients with stage IB1 squamous cell carcinoma of the uterine cervix.

p21 gene codon 31 arginine/serine polymorphism: non-association with endometriosis.

p21, an important regulator of the cell cycle, acts as a mediator of the growth-suppressing and -promoting functions of p53. We aimed to investigate the association between codon 31 polymorphisms of p21 gene and endometriosis. Women were divided into two groups: endometriosis (n = 102) and nonendometriosis (n = 119). The gene polymorphism for p21 codon 31 involved a base change from AGC to AGA and amino acid changes from serine (Ser) to arginine (Arg). Polymorphisms (Ser homozygotes, heterozygotes, Arg homozygotes) between both groups were detected and compared. Associations between the endometriosis and polymorphisms were evaluated. The results revealed that the distributions of different p21 polymorphisms in both groups were nonsignificantly different. The proportions of Ser homozygote/heterozygote/Arg homozygote in endometriosis and nonendometriois populations were 26.5/48.0/25.5% and 17.6/50.4/31.9%, respectively. We concluded the noncorrelation between the endometriosis and the p21 codon 31 polymorphism. p21 gene codon 31 arginine/serine polymorphism is not a useful marker for prediction of endometriosis susceptibility.

Effect of methylglyoxal on mouse embryogenesis.

BACKGROUND: Methylglyoxal (MG), a highly active and mutagenetic compound, has been found widely in a variety of foods and beverages. We investigated the effect of MG on mouse embryo development in-vitro. METHODS: Two-cell mouse embryos were divided into six groups according to the MG concentration in the culture medium: Group 1 (control group). 0 mM; Group 2, 10(-4) mM; Group 3, 10(-3) mM; Group 4, 10(-2) mM; Group 5, 10(-1) mM; Group 6, 1 mM. Embryo development and cleavage were compared every day for 5 days. RESULTS: The percentages of embryos reaching blastocyst/hatching stages were as follows: Group 1, 66.8%/34.2%; Group 2, 67.9%/38.7%; Group 3, 56.2%/31.5%; Group 4, 39.4%/14.1%; Group 5, 11.4%/10.2%; Group 6, 0%/0%. Higher MG concentrations (> or = 10(-2) mM) were associated with morphological aberrations and blocked development of embryos. CONCLUSION: The cutoff value of MG concentration on the mouse embryo development in-vitro is 10(-2) mM. An increased risk of embryotoxicity occurs with MG concentrations > or = 10(-2) mM in vitro. There were no significant effects on the growth rate at MG concentrations of 10(-3) and 10(-4) mM.

Prenatal diagnosis of oculocutaneous albinism two mutations located at the same allele.

A pregnant woman accepted amniocentesis on account of the previous birth of type 1 oculocutaneous albinism (OCA1). PCR revealed that the fetus had two mutations (862delTT, Arg 299His). The father had one missense mutation (Arg 299Ser) and the mother had the same mutations as the fetus. Two mutations of the fetus located at the same allele were suspected. Postpartal follow-up confirmed his carrier status. For recessive disorders, faced with a fetus with two mutations, the importance of performing segregation analysis of mutation on both parents is emphasized. This could exclude two mutations located at the same allele and prevent the unnecessary termination of a fetus with carrier status.

Longitudinal survey of blood flow at three different locations in the middle cerebral artery in normal fetuses.

OBJECTIVES: To assess vascular impedance at three different locations in the middle cerebral artery (MCA) in normal fetuses throughout gestation. MATERIALS AND METHODS: Uncomplicated singleton pregnancies at 15-40 weeks' gestation in which Doppler surveys of both MCAs could be obtained were recruited. The pulsatility index (PI) and resistance index (RI) of the proximal, mid and distal sites of both MCAs were measured. The five gestation periods at which the Doppler surveys were performed were (i) 15-19 completed weeks; (ii) 20-24 completed weeks; (iii) 25-29 completed weeks; (iv) 30-34 completed weeks; (v) 35-39 completed weeks. RESULTS: A total of 42 fetuses were recruited. The average PI/RI values of the proximal, mid and distal MCA were 1.61/0.82, 1.77/0.82, and 1.84/0.84, respectively. The PI value of the proximal MCA was lower than that of the mid and distal MCA. The PI values of the mid and distal MCA showed no difference. The RI value of the MCA from the three locations also showed no difference. The PI/RI values of MCA for each gestational phase were: (i) 1.81/0.87; (ii) 1.79/0.86; (iii) 1.78/0.86; (iv) 1.70/0.81; (v) 1.62/0.77, respectively. Decreased PI/RI values were observed after 30 weeks' gestation. CONCLUSIONS: The PI values of the proximal MCA are lower than those of the mid and distal MCA. A marked decrease in PI/RI values was observed after 30 weeks' gestation.

Leukemia inhibitory factor decreases the arylamine N-acetyltransferase activity in human cumulus granulosa cells.

PURPOSE: To evaluate the activities of acetyl coenzyme A (AcCoA):arylamine N-acetyltransferase (NAT) of intact cumulus granulosa cells and the role of leukemia inhibitory factor (LIF) upon their NAT activities. METHODS: Thirty women accepted controlled ovarian hyperstimulation (COH) and oocyte retrievals. Human cumulus granulosa cells were obtained during oocyte retrievals. Using 2-aminofluorene (2-AF) and p-aminobenzoic acid (PABA) as substrates, NAT activity of all samples was determined by high pressure liquid chromatography. After the incubation with different time and concentrations of 2-AF, PABA, and LIF, 2-acetyl-aminofluorene (2-AAF) and N-acetyl-PABA (N-Ac-PABA) were measured. RESULTS: After incubation with 2.812, 5.625, 11.25, and 22.5 microM of 2-AF/PABA, their product concentrations (2-AAF/N-Ac-PABA) were 0.42/0.32, 0.76/0.58, 1.29/1.04, and 1.94/1.26 nmol/10(6) cells, respectively. After 6, 12, 18, and 24 h incubation with 11.25 microM of 2-AF/PABA, their product concentrations were 0.19/0.12, 0.56/0.4, 0.98/0.79, and 1.3/1.0 nmol/10(6) cells, respectively. After incubation with 0, 5, and 50 microM of LIF, the 2-AAF/N-Ac-PABA concentrations were 0.98/0.80, 0.70/0.52, and 0.49/0.30 nmol/10(6) cells, respectively. CONCLUSION: Intact human cumulus granulosa cells could acetylate arylamine carcinogen (2-AF) and noncarcinogens drug (PABA). LIF decreased the NAT activities. It provides a model for monitoring the effects of COH and LIF upon the oocytes.

Doppler evaluation of the uterine and spiral arteries from different sampling sites and phases of the menstrual cycle during controlled ovarian hyperstimulation.

OBJECTIVE: To compare the pulsatility index (PI) and resistance index (RI) at different sampling sites of the uterine and spiral arteries in the early and mid-menstrual phases. MATERIALS AND METHODS: The uterine and spiral arteries of 110 women undergoing similar ovarian hyperstimulation and intra-uterine insemination regimes were examined using transvaginal color Doppler ultrasound. The uterine artery was sampled at five sites: (1) ascending branch; (2) descending branch; (3) proximal, near branch division; (4) mid, 0.5 cm distal to the division; (5) lateral location, 1 cm distal to the division. The spiral artery was sampled at three sites: (a) anterior; (b) fundal; (c) posterior. The uterine and spiral arteries were examined twice, on days 2-3 and 14-16, respectively, of the menstrual cycle. The women were also grouped according to age at examination, < or = 30 years and > 30 years. The PI and RI values for different sites, menstrual phase, and age were compared. RESULTS: The mean PI and RI values of the uterine artery were: (1) 2.86 +/- 1.20 and 0.92 +/- 0.13; (2) 2.66 +/- 1.15 and 0.89 +/- 0.12; (3) 2.88 +/- 1.26 and 0.90 +/- 0.15; (4) 3.03 +/- 1.02 and 0.91 +/- 0.07; (5) 3.23 +/- 1.38 and 0.89 +/- 0.12; and of the spiral artery were (a) 1.61 +/- 1.01 and 0.69 +/- 0.17; (b) 1.69 +/- 0.74 and 0.74 +/- 0.17; (c) 1.73 +/- 0.86 and 0.68 +/- 0.17. The PI values for uterine and spiral arteries at two phases of the menstrual cycle were 2.92 +/- 1.18 and 1.55 +/- 0.72 (days 2-3); 3.11 +/- 1.15 and 1.80 +/- 1.02 (days 14-16), respectively; for younger women (age < or = 30 years) these values were 2.83 +/- 1.22 and 1.6 +/- 0.85 and for older women (age > 30 years) 3.0 +/- 1.34 and 1.72 +/- 0.96, respectively. CONCLUSIONS: There were no significant differences in PI and RI values of the uterine and spiral arteries at different sampling sites, phase of the menstrual cycle or age. The higher PI values tended to occur in the lateral uterine artery and posterior spiral artery, during the mid-menstrual phase and in the older age group. The PI and RI values of the mid-uterine and fundal spiral artery sampling sites are representative of the whole uterine artery and spiral artery, respectively.

Accuracy of sonography in predicting the outcome of fetal congenital diaphragmatic hernia.

BACKGROUND: The outcome of congenital diaphragmatic hernia (CDH) remains poor despite recent advances in neonatal care. This study was designed to evaluate the role of sonography in predicting the outcome of CDH. METHODS: Pregnancies with CDH were studied. Fetal survival, morbidity, combined anomalies and mortality were recorded. Seven parameters were recorded, including the presence of hydramnios, side of herniation, cardiac deviation, stomach presence, gestational age at the time of finding the CDH and time of postpartum herniorrhaphy. The predictive values of these parameters for fetal outcome were analyzed. RESULTS: A total of 31 pregnancies were studied. There were 11 cases (35.5%) of termination, seven cases (22.6%) of perinatal death, four cases (12.9%) of late death and nine cases of survival (29%). The survivor group included four cases (44.4%) of complete recovery and five cases (55.6%) with persistent morbidity. There were 15 cases of simple CDH including eight cases of cardiac anomalies (ventricular-septal defect, atrial-septal defect, patent ductus arteriosus and ventricular dilatation). There were eight cases with severe anomalies (3 with trisomy 18, 2 with Cantrell's pentalogy, 1 with trisomy 13, 1 with cystic hygroma and one with Tetralogy Fallot). Among the seven parameters studied, gestational age at the time of finding the CDH and hydramnios were related to fetal survival. CONCLUSIONS: Sonography assists in predicting the postnatal outcome of CDH. Diagnosis of CDH at less than 25 weeks' gestation and the existence of hydramnios are associated with higher mortality. Postnatal therapy and prenatal surgical intervention are necessary to salvage fetuses in the presence of these two situations. The survival rate of infants with CDH was 45%. Of these, 55.6% had persistent morbidity. Prenatal counseling should reflect this.

Routine blastocyst culture and transfer: 201 patients' experience.

PURPOSE: The purpose was to compare the outcomes between day-5 blastocyst and day-2 embryo transfers. METHODS: Infertile women who accepted the ovarian hyperstimulation and oocyte retrieval were divided: Group 1, day 2 embryo transfer, group 2, cultured to day 5 in serum-free sequential culture medium and transfer. Early embryo quality and growth, blastocyst formation and quality, implantation rate (IR) and pregnancy rates (PR) were detected. RESULTS: Total blastocyst formation rate was 49.4%. Better early embryo quality (days 2, 3) and higher day 3 blastomere number possess higher blastocyst formation rate. The IR for day 2 and day 5 embryos were 10.8% and 22.2%, respectively. The PR in both groups were comparable (37.3% vs. 41.8%). CONCLUSIONS: Blastocyst transfer has higher IR and comparable PR as those of day 2 embryo transfer. Early embryo qualities and day 3 blastomere number are useful in predicting the final blastocyst formation. Blastocyst formation rate is not related to maternal age, infertile causes, insemination methods, and early embryo number.

Leukemia inhibitory factor expression in different endometrial locations between fertile and infertile women throughout different menstrual phases.

PURPOSE: The purpose was to demonstrate the leukemia inhibitory factor (LIF) expression in different endometrial locations between fertile and infertile women throughout different menstrual phases. The relationship between progesterone level and LIF expression were evaluated. METHODS: Endometrial biopsies were performed on idiopathic infertile and normal fertile women accepted the in follicular, periovulatory, and luteal phases. The luteal progesterone level was measured. Endometrial LIF immunostaining of luminal epithelium, glandular epithelium, and stroma were detected. The relationship between luteal LIF expression and progesterone level was evaluated. RESULTS: Significant LIF expression was noted in the endometrium of fertile women rather than that of infertile women. The LIF expression was highest in the luminal epithelium, moderate in the glandular epithelium, and lowest in the stroma. The luminal and glandular epithelial staining were lowest in follicular phase, moderate in periovulatory phase, and strongest in luteal phase. The stromal LIF presented with a noncyclical manner. The LIF expression is not related with the progesterone level. CONCLUSIONS: Endometrial LIF expression is related to human fertility. Endometrial LIF expression is dependent on cellular localizations and menstrual stages. Stronger LIF expression presents in the endometrial epithelium during luteal phase.