Differential motility parameters and identification of proteomic profiles of human sperm cryopreserved with cryostraw and cryovial.

BACKGROUND: Although sperm cryopreservation has been widely used in human reproductive medicine as an integral infertility management in infertility clinics and for banking sperm in sperm banks, the freezing/thawing protocols are not optimal. The freezing and thawing processes result in changes at both structural and molecular levels, some even detrimental, in human sperm when compared with fresh sperm. The change of sperm proteins after cryopreservation may play negative roles for fertilization and early embryo development. Conventionally, cryostraws (CS) and cryovials (CV) are the most widely used cryopreservation carriers (CPCs) for human sperm cryopreservation accompanied with the use of egg yolk free commercial media. However, the influence of cryopreservation on the proteomic profile of human sperm preserved with the two CPCs is unknown. Therefore the purpose of the present study was to compare the frozen-thawed motility, investigate the proteomic profile of human sperm cryopreserved with the two types of CPCs, and identify the susceptible proteins that play key roles for sperm function and fertility. METHODS: The present study compared the cryosurvival of human sperm frozen with the two different CPCs and identified the sperm proteomic changes by using the isobaric tags for relative and absolute quantification labeling technique coupled with 2D LC-MS/MS analysis after freezing and thawing. RESULTS: Our results indicated that sperm cryopreserved with CV showed higher values for percentage of motile sperm and forward activity rate than those with CS. Compared to fresh sperm, 434 and 432 proteins were differentially identified in human sperm cryopreserved with CS and CV, respectively. CONCLUSION: The proteomic profiles of human sperm are greatly affected by cryopreservation with either type of CPC. GO analysis revealed that most of the differentially identified sperm proteins enriched in the extracellular membrane-bounded organelles, cytoplasm and cytosol. In addition, 106 susceptible proteins having known identities related to sperm functions were identified. In general, cryovial seems to be the preferred CPC for human sperm cryopreservation based on the post-thaw motility parameters and the effect on sperm proteomic profiles. These results are beneficial for the insight into the understanding of the cryoinjury mechanism of sperm and the development of human sperm cryopreservation strategies.

Does reducing gamete co-incubation time improve clinical outcomes: a retrospective study.

PURPOSE: The objective of this retrospective study was to determine whether patients undergoing in vitro fertilization (IVF) benefit from reducing the gamete co-incubation time. METHODS: Patients (n = 570) were enrolled, including 281 patients in the reduced incubation time group (2-h incubation) and 289 patients in the standard IVF group (18-h incubation). RESULTS: The observed outcomes, including the clinical pregnancy rate (CPR), implantation rate (IR), live birth rate (LBR), and miscarriage rate (MR), were similar between the two groups. When the data were divided into two subgroups based on the maternal age (≤30 and >30 years), the rates of top-quality embryos (30.83 vs. 25.89 %; p = 0.028), CPR (66.67 vs. 42.11 %; p = 0.013), and IR (41.90 vs. 31.25 %, p = 0.019) of the 2-h incubation group were significantly higher in the younger subgroup. However, for older patients, only a lower MR (7.59 vs. 20.83 %; p = 0.019) was achieved. Reducing the time of incubation still improved the CPR (OR = 1.993, 95 % CI 1.141-3.480) and MR (OR = 3.173, 95 % CI 1.013-9.936) in the younger and older subgroups, respectively, after it was adjusted for potential confounders. CONCLUSIONS: Reducing incubation time improves the clinical results of IVF, although the LBR is not statistically different between the 2- and 18-h incubation time groups. And the specific clinical outcomes of reducing incubation time varied between the >30-year-old and the ≤30-year-old.

[Study on the facial and body terminal hair growth in women in Guangdong by using modified Ferriman-Gallwey scoring system].

OBJECTIVE: To study growth of facial and body terminal hair of women in Guangdong province and its relationship with age, menstrual irregularities and polycystic ovary, and determine normative cut-off score of modified Ferriman and Gallwey (mFG). METHODS: A cross-sectional study was conducted on 2988 women at age of 20-45 years from 16 communities of two urban and two rural regions in Guangdong province from June 2008 to July 2009. Terminal body hair growth was assessed by using the modified Ferriman and Gallwey (mFG) scoring system. The normative cut-off value of mFG were calculated by using the K-means cluster analysis (K=2). Those women were classified into following groups, including 982 women at group of ages of 20- years, 765 women at group of 26- years, 597 women at group of 31- years, 384 women at group of 36- years, 260 women at group of 41-45 years. Due to absence or errors of medical records, some cases were excluded from this study. Based on menses irregularities (MI), polycystic ovaries (PCO), there were 488 cases in MI group, 2413 cases in normal menses group, 568 cases in PCO group, and 2207 cases in non-PCO group finally. The incidences of acne, MI, acanthosis nigricans, and polycystic ovaries were also analyzed in all the hirsute groups. RESULTS: (1) among 2988 women, it was observed 149 women (5%) with mFG≥7,314 women (10.5%) with ≥5,747 women with mFG≥2. (2) Cluster analysis identified an mFG score of 5 as the cut-off value that define abnormal hirsute in the total population and all the sub-groups with/without MI or PCO; (3) Based on age classification, it was found that increased age was associated with decreased trends of the percentile and cut-off value of hirsutism. The value of hirsutism of mFG were 6 in group of 20- years, 5 in group of 26- years, 4 in groups of 31- years, 36- years and 41-45 years. (4) The prevalence of acne, menstrual irregularities and POC were 45.5% (143/314), 73.6% (231/314), 25.8% (81/314) in total population, 25.1% (671/2674), 16.1% (431/2674), 19.8% (529/2674) in normal hair women, which reached statistical difference (P<0.05). The prevalence of acne, menstrual irregularities and acanthosis nigricans were 44.4% (130/293), 23.2% (68/293), 4.1% (12/293) in those age hirsute groups, 25.3% (681/2695), 16.2% (437/2695), 1.9% (51/2695) in normal hair women, which reached statistical difference (P<0.05). CONCLUSIONS: (1) among women in Guangdong province, mFG scoring showed decreased trends in women with increasing age. (2) An mFG score≥5 was cut-off value in diagnosis of hirsutism. (3) The hirsute women exhibited higher incidence of acne, menses irregularity, and acanthosis nigricans than those of women with normal hair growth.

Endocrine profiles and cycle characteristics of infertile 17α-hydroxylase/17,20-lyase Deficiency Patients undergoing assisted Reproduction Treatment: a retrospective cohort study.

BACKGROUND: 17α-hydroxylase/17,20-lyase deficiency (17-OHD) is a rare form of congenital adrenal hyperplasia caused by CYP17A1 gene variants. Female patients with 17-OHD demonstrate a broad clinical spectrum, including oligomenorrhea or amenorrhea and infertility, often as the sole manifestation. However, no spontaneous pregnancies in affected women have been reported. OBJECTIVE: This retrospective cohort study aimed to explore the endocrine characteristics and assisted reproductive technique (ART) performance in women with 17-OHD. METHODS: Five women were referred for primary infertility in a university-affiliated hospital over an eight-year period. The endocrine profiles and cycle characteristics during a total of nine cycles of ovarian stimulation and eight cycles of frozen-thawed embryo transfer (FET) were described in details. RESULTS: Three cases had homozygous variants and two cases had compound heterozygous variants, including one novel missense variant (p.Leu433Ser) in the CYP17A1 gene. Despite dual-suppression of progesterone (P) production by glucocorticoid and gonadotropin releasing hormone agonist, gradually increased P level, relatively low estradiol concentrations and thin endometrium were observed, negating fresh embryo transfer. During FET cycles, appropriate treatment resulted in low serum P levels and adequate endometrial thickness, leading to four live births. CONCLUSIONS: Our findings demonstrate that continuous elevation of serum P during follicular growth impairs endometrial receptivity, the likely cause of female infertility in 17-OHD. Therefore, female infertility caused by 17-OHD is suggested as an indication for freeze-all strategy, with promising reproductive prognoses following segmented ovarian stimulation and FET treatment.

Defining hyperandrogenaemia according to the free androgen index in Chinese women: a cross-sectional study.

OBJECTIVE(S): To determine the normative cut-off value for the free androgen index (FAI) among reproductive-age women in China. DESIGN: Cross-sectional, population-based study. SETTING: A multistage systematic cluster random sampling among 16 communities from two rural and two city regions. PATIENT(S): A total of 1526 women were randomly selected from a representative sample of 3009 women aged 20-45 years from the general population of southern China. Among these women, 444 subjects without known factors affecting androgen levels were selected as the reference group. INTERVENTION(S): Subjects underwent physical examination, blood tests of hormone and metabolic profiles, and ultrasound evaluations. MEASUREMENT OF THE MAIN OUTCOMES: The FAI was calculated according to the following equation: FAI = 100 × total testosterone (nmol/l)/sex hormone-binding globulin (nmol/l). The normative cut-off value of the FAI was computed based on the 5th-95th percentiles and K-means cluster analysis (K = 2). RESULTS: A reference range of 0·7-6·4 of the FAI was indicated by the 5th and 95th percentiles. K-means cluster analysis identified 6·1 as the cut-off value for the FAI in the total population, which was close to the upper limit determined by the 95th percentile. Hyperandrogenic women present with elevated risk factors for metabolic disorders. The mean FAI values decreased with age among the total population, as evaluated by either K-means clustering or a percentile analysis. The FAI value was negatively correlated with age (r = -0·185, P < 0·0001) and positively correlated with body mass index after adjustment for age (r = 0·250, P < 0·0001). CONCLUSION: A reference range of 0·7-6·4 FAI was indicated by the 5th and 95th percentiles among the reference group, which excluded the subjects with factors known to affect androgen levels; 6·1 was identified by the K-means cluster analysis as the cut-off value of the FAI for the general population. Hyperandrogenic women present with higher risks for metabolic disorders.

Appropriate BMI levels for PCOS patients in Southern China.

BACKGROUND: This study was designed to establish appropriate body mass index (BMI) level for PCOS women and to compare metabolic abnormalities between PCOS women and control women in Southern China. METHODS: This clinical cross-sectional study included 999 PCOS patients (857 adults and 142 adolescents) and 922 age-matched controls (742 adults and 180 adolescents). RESULTS: The proportion of PCOS patients with a BMI above 23 kg/m(2) was 34.63%. Serum insulin, triglycerides, waist and waist/hip (W/H) measurements were all correlated positively with BMI in PCOS women. Prevalence of metabolic syndrome in Chinese PCOS patients was 18.9%. Receiver-operating characteristic analysis suggested that at the point of BMI > or =23 kg/m(2), the diagnostic power to detect metabolic disorders was at its best. CONCLUSIONS: PCOS patients with a BMI of 23 kg/m(2) or beyond may have a higher risk of metabolic disorders. Using an appropriate BMI level may help to improve attention to metabolic disorders among PCOS patients in Southern China.

Metabolic abnormalities in adolescents with polycystic ovary syndrome in south China.

BACKGROUND: Adults with polycystic ovary syndrome (PCOS) can have multiple metabolic abnormalities. However, studies in the adolescent population are still limited and these results seem to vary widely. This study was to investigate the metabolic abnormalities in adolescents with PCOS in South China and the potential risk factors contributed to these health risks. METHODS: Anthropometric measurements and biochemical parameters were evaluated in 128 adolescents with PCOS and their age- and BMI-matched controls. RESULTS: The prevalence of pre-diabetes, insulin resistance, hyperinsulinemia, dyslipidemia and metabolic syndrome in adolescents with PCOS was 11.7%, 46.9%, 29.7%, 22.7% and 4.7%, respectively. 16.3%, 74.4%, 67.4%, 39.5% and 14% of the PCOS subjects with BMI > 85th had pre-diabetes, insulin resistance, hyperinsulinemia, dyslipidemia and metabolic syndrome, whereas 9.4%, 32.9%, 10.6%, 14.1% and 0% of the PCOS subjects with BMI < 85th had such disturbances. CONCLUSIONS: Adolescents with PCOS in South China had more metabolic abnormalities than their age- and BMI-matched non-PCOS counterparts. Obesity could worsen insulin resistance, hyperinsulinemia and metabolic syndrome in PCOS adolescents.

Statistical Genomic Approach Identifies Association between FSHR Polymorphisms and Polycystic Ovary Morphology in Women with Polycystic Ovary Syndrome.

BACKGROUND: Single-nucleotide polymorphisms (SNPs) in the follicle stimulating hormone receptor (FSHR) gene are associated with PCOS. However, their relationship to the polycystic ovary (PCO) morphology remains unknown. This study aimed to investigate whether PCOS related SNPs in the FSHR gene are associated with PCO in women with PCOS. METHODS: Patients were grouped into PCO (n = 384) and non-PCO (n = 63) groups. Genomic genotypes were profiled using Affymetrix human genome SNP chip 6. Two polymorphisms (rs2268361 and rs2349415) of FSHR were analyzed using a statistical approach. RESULTS: Significant differences were found in the allele distributions of the GG genotype of rs2268361 between the PCO and non-PCO groups (27.6% GG, 53.4% GA, and 19.0% AA versus 33.3% GG, 36.5% GA, and 30.2% AA), while no significant differences were found in the allele distributions of the GG genotype of rs2349415. When rs2268361 was considered, there were statistically significant differences of serum follicle stimulating hormone, estradiol, and sex hormone binding globulin between genotypes in the PCO group. In case of the rs2349415 SNP, only serum sex hormone binding globulin was statistically different between genotypes in the PCO group. CONCLUSIONS: Functional variants in FSHR gene may contribute to PCO susceptibility in women with PCOS.

Defining hirsutism in Chinese women: a cross-sectional study.

OBJECTIVE: To determine the normative cut-off score that defines hirsutism among Chinese women, and the effect of age, menstrual irregularities, and polycystic ovaries on the same. DESIGN: Cross-sectional, population-based study. SETTING: A multistage systematic cluster random sampling among 16 communities from two rural and two city regions. PATIENT(S): A representative sample of 2,988 women aged 20-45 years from the general population of Southern China. INTERVENTION(S): Subjects underwent physical and ultrasound evaluations. MAIN OUTCOME MEASURE(S): Terminal hair growth was assessed using the modified Ferriman-Gallwey (mFG) scoring system. RESULT(S): An mFG score ≥5 was observed in 10% and a score of ≥2 in 25% of the subjects. Cluster analysis identified an mFG score of 5 as the cut-off value that defines abnormal terminal facial and body hair growth in a male pattern (i.e., hirsutism) in the total population; scores of 6, 5, and 4 for women aged 20-25, 26-30, and >30 years, respectively. Defined by these cut-off values, the prevalence of hirsutism in our total population was 10.5%; and decreased with increasing age: 14.4%, 10.7%, 7.9%, 3.6%, and 1.5%, respectively, in women aged 20-25, 26-30, 31-35, 36-40, and 41-45 years. Furthermore, the incidence of acne, menses irregularities, polycystic ovaries, and acanthosis nigricans were significantly increased among the hirsute women. CONCLUSION(S): An mFG score of 5 or greater indicates hair growth above the norm among women in the general Southern Chinese population, a cut-off value that decreases with increasing age.

Low prevalence of the metabolic syndrome but high occurrence of various metabolic disorders in Chinese women with polycystic ovary syndrome.

OBJECTIVE: Variations in the prevalence of metabolic syndrome (MetS) among women with polycystic ovary syndrome (PCOS) in different races were reported. We sought to report this prevalence and its components in Chinese women with PCOS and compared these characteristics with healthy controls. DESIGN: Anthropometric measurements and biochemical parameters were evaluated in 578 PCOS patients diagnosed by the Rotterdam criteria and 281 age- and body mass index (BMI)-matched controls. International Diabetes Federation criteria for MetS were used. RESULTS: The prevalence of MetS was 16.8% in this study, and 60.7% of patients displayed at least one component of MetS. Among the patients, the rates of dyslipidemia, impaired fasting glucose, and elevated blood pressure were 41.6, 19.8, and 16.1% respectively; the rates of these corresponding components in age- and BMI-matched controls were 14.6, 5.3, and 5.7% respectively. In PCOS patients, the prevalence of MetS was 0.0, 3.9, 20.2, and 51.1% for four different BMI groups respectively; the prevalence of MetS was 7.3, 14.9, 24.2, and 42.4% in the four age groups respectively. Nearly 90% of patients diagnosed with MetS belonged to overweight and obese groups. BMI and age rather than free testosterone, free androgen index, fasting insulin, or sex hormone-binding globulin were included in formulation for predicting MetS according to multivariable logistic regression. CONCLUSIONS: Low prevalence of MetS but high occurrence of various metabolic disorders was found in women with PCOS compared with age- and BMI-matched controls in this study. BMI and age appeared to contribute more to developing MetS than other parameters associated with insulin resistance or hyperandrogenism.