BBOF1 is required for sperm motility and male fertility by stabilizing the flagellar axoneme in mice.

The sperm flagellum is a specialized type of motile cilium composed of a typical "9 + 2" axonemal structure with peri-axonemal structures, such as outer dense fibers (ODFs). This flagellar arrangement is crucial for sperm movement and fertilization. However, the association of axonemal integrity with ODFs remains poorly understood. Here, we demonstrate that mouse BBOF1 could interact with both MNS1, an axonemal component, and ODF2, an ODF protein, and is required for sperm flagellar axoneme maintenance and male fertility. BBOF1 is expressed exclusively in male germ cells from the pachytene stage onwards and is detected in sperm axoneme fraction. Spermatozoa derived from Bbof1-knockout mice exhibit a normal morphology, however, reduced motility due to the absence of certain microtubule doublets, resulting in the failure to fertilize mature oocytes. Furthermore, BBOF1 is found to interact with ODF2 and MNS1 and is also required for their stability. Our findings in mice suggest that Bbof1 could also be essential for human sperm motility and male fertility, thus is a novel potential candidate gene for asthenozoospermia diagnosis.

Clinical outcomes of fresh and frozen embryos transfer after in vitro fertilization in spouses of patients with severely low sperm count and motility. / å› é å¶é‡åº¦å°‘å¼±ç²¾å­ç—‡ä¸å­•æ‚£è€ ä½“å¤–å—ç²¾åŽç§»æ¤æ–°é²œèƒšèƒŽä¸Žå†·å†»èƒšèƒŽçš„ä¸´åºŠç»“å±€æ¯”è¾ƒ.

OBJECTIVES: To compare the pregnancy and neonatal outcomes of in vitro fertilization-embryo transfer (IVF-ET) with fresh or frozen embryos for male patients with severely low sperm count and motility. METHODS: A total of 2300 male patients with severely low sperm count and motility underwent IVT-ET in the Reproduction Medicine Center, Sir Run Run Shaw Hospital from April 2018 to April 2022. After applying the propensity score matching (PSM), 473 fresh embryo transferred cycles and 473 frozen embryo transferred cycles were selected in the study, and the pregnancy and neonatal outcomes were compared between two groups. RESULTS: There were no significant differences in pregnancy outcomes and neonatal outcomes between fresh and frozen embryo groups (all P>0.05). In the stratification analysis, the number of retrieved oocytes in the fresh good-quality embryo transfer group was significantly increased compared with the fresh poor-quality embryo group (P<0.05), but the very early pregnancy loss rates were similar between the two groups, while the rate in fresh good-quality embryo transfer group was significantly higher than that in the frozen good-quality embryo transfer group (P<0.05). Among different age groups of women, the number of retrieved oocytes and the level of estrogen in the fresh embryo transfer group was significantly higher in the 20 to <30 years old group than that in the 30 to <35 years old group (both P<0.05), but the clinical pregnancy rate was lower in the 20 to <30 years old group than that in the 30 to <35 years old group (P>0.05). Additionally, the very early pregnancy loss was significantly increased in the fresh embryo group compared with the frozen embryo group in the 20 to <30 years age group (P<0.05). CONCLUSIONS: There is no significant difference in pregnancy and neonatal outcomes between fresh embryo transfer and frozen embryo transfer for male patients with severely low sperm count and motility undergoing IVF-ET. Due to shorter transfer time, less embryo freezing damage and reduced costs, fresh embryo transfer can be considered a first choice. However, it is not necessary to pursue fresh embryo transfer if maternal oestrogen levels are too high and there is a tendency of overstimulation.

A validated high-performance liquid chromatography method for detecting geniposide, ellagic acid, piperine, costunolide and dehydrocostus lactone in Liuwei Muxiang capsules.

In this study, a sensitive high-performance liquid chromatography detector was established and validated for the simultaneous determination of geniposide, ellagic acid, piperine, costunolide and dehydrocostuslactone in Liuwei Muxiang Capsules. The analysis was achieved on CHANIN 100-5-C18-H column (5µm, 250 mm×4.6 mm) with the temperature of 30oC. Gradient elution was applied using 0.1% phosphoric acid solution-methanol-acetonitrile (50:50) as mobile phase at the flow rate of 1.0 mL/min. The determination was performed at the wavelength of 225 nm (detecting geniposide), 254 nm (detecting ellagic acid), 343 nm (detecting piperine) and 225 nm (detecting costunolide and dehydrocostuslactone) along with the sample volume of 10µL. The linear ranges of geniposide, ellagic acid, piperine, costunolide and dehydrocostuslactone demonstrated good linear relationships within their respective determination ranges. The average recoveries were 100.04%, 99.86%, 99.79%, 100.17% and 100.41%, respectively. RSD% was 1.3%, 1.2%, 1.2%, 1.2%, 1.5%, respectively. The developed method was proved to be simple, accurate and sensitive, which can provide a quantitative analysis method for the content determination of geniposide, ellagic acid, piperine, costunolide and dehydrocostuslactone in Liuwei Muxiang capsules.

Clinical outcomes of frozen-thawed blastocysts from zygotes with no or one pronucleus for in vitro fertilization and intracytoplasmic sperm injection cycles.

PURPOSE: To investigate the clinical outcomes of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles using frozen-thawed blastocyst transfers derived from zygotes with no (0PN) or one pronucleus (1PN). METHODS: This retrospective study included 7084 0PN, 2238 1PN, and 72,266 two pronuclear (2PN) embryos cultured to the blastocyst stage from 19,631 IVF and 12,377 ICSI cycles between March 2018 and December 2021. Developmental potential and clinical outcomes of 0PN, 1PN, and 2PN embryos were analyzed. A total of 290 0PN-, 92 1PN-, and 1906 2PN-derived single frozen-thawed blastocyst transfers were performed. Chromosome euploid rates of 0PN-, 1PN-, and 2PN-derived blastocysts were analyzed by next-generation sequencing. Euploid 0PN- and 1PN-derived blastocysts underwent subsequent Infinium Asian Screening Array gene chip analysis to detect ploidy alterations. RESULTS: Available blastocyst rates of 0PN and 1PN embryos were significantly lower than those of 2PN embryos in both IVF and ICSI cycles. Single 0PN and 1PN blastocysts transferred in frozen-thawed cycles resulted in a similar clinical pregnancy rate, miscarriage rate, live birth rate, and neonatal outcome to 2PN blastocysts in IVF and ICSI cycles. Genetic analysis showed that euploid rates of 0PN- and 1PN-derived blastocysts used for ICSI cycles were similar to that of 2PN-derived blastocysts. CONCLUSION: Our study indicated that 0PN- and 1PN-derived blastocysts resulted in similar clinical outcomes to 2PN-derived blastocysts. The 0PN- and 1PN-derived blastocysts from ICSI cycles can be transferred as well as those from IVF cycles when the number of 2PN-derived blastocysts is insufficient.

A novel solution for freezing individual spermatozoa using a right angular cryopiece embedded in a grooved petri dish.

Herein, we introduced a novel individual sperm freezing device named SpermCD, which consists of a right angular cryopiece (RA-Cryopiece, or "C") and a grooved petri dish ("D"). SpermCD allows embryologists to transfer sperm and perform ICSI on the same focal plane. Thirty-five patients underwent single sperm cryopreservation using SpermCD, including four patients with non-obstructive azoospermia (NOA), 14 patients with virtual azoospermia and 17 patients with cryptozoospermia. One hundred and twenty-five cryopreserved spermatozoa from nine patients were thawed on the day of the oocyte retrieval and 121 spermatozoa were found, with a sperm recovery rate of 97.1 ± 4.6%. Sixty-five MII oocytes from their spouse were injected with thawed sperm. Normal fertilization and high-quality embryo rates were 68.0% ± 33.2% and 24.4% ± 22.2%. Nineteen transplantable embryos were formed after fertilization with frozen sperm, eight of which were transplanted in five couples, resulting in four successful deliveries. SpermCD is a simple and practical individual sperm freezing device.

Effects of auxiliary atmospheric state parameters on the aerosol optical properties retrieval errors of high-spectral-resolution lidar.

A detailed assessment is carried out in relation to the influence of the uncertainties associated with the input auxiliary atmospheric state parameters on retrieving aerosol optical properties from high-spectral-resolution lidar (HSRL) observations. The study starts from a review of the main spectral structure of the Rayleigh backscattering followed by evaluating the temperature effects on a backscattering cross section of atmospheric molecules based on numerical simulation. It shows that the transmittance of the background interference filter should be taken into account, depending on the full width at half maximum, although overall temperature dependence is negligible. Based on the Taylor expansion of the Tenti S6 model, the systematic errors arising from input temperature and pressure profiles are analyzed. It is demonstrated that the atmospheric pressure profiles have limited effects on the inversion results of aerosol optical parameters, as the atmospheric pressure is usually quite stable. The relative errors of the aerosol backscatter coefficient mainly stem from temperature profile errors and highly depend on the aerosol concentration. Quantitatively, the aerosol backscatter coefficient error could be larger than 5% with a 3 K deviation of temperature when the backscatter ratio is larger than 1.1. The accuracy of aerosol extinction coefficient retrieval is affected not only by the error in temperature, but also by the error in temperature lapse rate; the retrieval accuracy is more sensitive to the latter than the former. Further analysis based on the sounding temperature data shows that the variation of the temperature inversion layer during the night could induce a bias larger than 0.04 km-1 on the aerosol extinction coefficient retrieval. Therefore, the time resolution of temperature measurement from sounding balloons twice per day is too low to obtain an accurate retrieval of the aerosol optical properties from the HSRL.

Effects of a nonideal half-wave plate on the gain ratio calibration measurements in polarization lidars.

A half-wave plate (HWP) is a critical component for calibrating the gain ratio in polarization lidars. In this paper, the effects of a nonideal HWP on the gain ratio calibration measurements are analyzed. We focus on the ±45° method and the multi-rotation HWP method, which are the two main approaches for calibrating the gain ratio. Specifically, we discuss the influences of a nonideal HWP from two scenarios: an HWP with nonideal polarization properties and an ideal HWP under nonideal external conditions. The allowable ranges of relevant parameters for the nonideal polarization properties of HWPs are obtained, which can help to determine qualified HWPs in polarization lidars. Several external conditions, including ambient temperature variations, wavelength differences, and the tilt angles of HWPs, are also analyzed in detail. To the best of our knowledge, this paper represents the first comprehensive study on the effects of nonideal HWPs on the gain ratio calibration measurements, offering some guidelines on choosing a qualified HWP for proper use in polarization lidars.

Slow freezing should not be totally substituted by vitrification when applied to day 3 embryo cryopreservation: an analysis of 5613 frozen cycles.

PURPOSE: This study aimed to compare slow freezing (SF) and vitrification (VT) techniques for day 3 embryo cryopreservation in infertile couples. METHODS: This retrospective cohort study enrolled 5613 infertile patients, with 7862 frozen-thawed day 3 embryos and 3845 vitrified-warmed day 3 embryos, from 2010 to 2014, at a single center. The rates of embryo survival, pregnancy, implantation, miscarriage, live birth, and live birth weight were compared between the two groups. RESULTS: A total of 5613 cycles with 5520 transfers were analyzed. Using SF, the rates of overall embryo survival and fully intact blastomeres were lower than those in VT (91.5 vs. 97.4 % and 68.7 vs. 92.3 %, respectively). The rate of good quality embryos after thawing/warming was lower in SF than in VT. In single frozen embryo transfer cycles (FETs), the pregnancy and implantation rates were similar between the two groups (35.0 vs. 40.8 % and 34.6 vs. 35.9 %, respectively). In double FETs, the pregnancy rate per cycle was also similar between the groups (58.8 vs. 58.4 %). The implantation rate per embryo transfer was significantly higher with SF than with VT (38.8 vs. 34.6 %). With adjustment for maternal age and the number of good quality embryos, differences in implantation rate remained significant (adjusted P value, SF vs. VT P < 0.05). No independent effect was found for the method of cryopreservation on the pregnancy rate. No significant differences in the rates of miscarriage, live birth, and live birth weight were observed between the two techniques. CONCLUSIONS: Despite the significantly low embryo survival rate, fully intact blastomere rate, and good quality embryo rate in SF, the pregnancy and implantation rates were not adversely affected in single and double FETs. SF yielded an equivalent miscarriage rate, live birth rate, and live birth weight compared with VT. The SF protocol to cryopreserve day 3 embryos still should be considered.

Effect of vitrification versus slow freezing of human day 3 embryos on ß-hCG levels.

PURPOSE: The study was designed to investigate the effect of vitrification and slow freezing for the cryopreservation of human day 3 embryos on serum ß-hCG levels in pregnancies established after frozen embryo transfer (FET). METHODS: Of the 1384 FET cycles initiated, 1131 embryo transfers met study criteria and assigned to one of two groups: 797 slow-freezing embryo transfers or 334 vitrified embryo transfers. Median values of ß-hCG and outcome of all pregnancies were compared between the two groups. Predictive values of serum ß-hCG on day 12 after embryo transfer for establishing ongoing pregnancy and pregnancy failure were determined by receiver operating characteristic (ROC) curve analysis. RESULTS: In the slow-freezing group, 383 ongoing pregnancies occurred (48.1 %), and transfers of vitrified embryos resulted in 154 pregnancies (46.1 %). Median ß-hCG values (279.2 IU/L) for ongoing pregnancies after transfer of vitrified embryos were significantly lower than that of slow frozen embryos (320.5 IU/L). The median values of ß-hCG for singleton in the two groups was statistically significant (P <0.05). For slow-freezing embryo transfers, the cut-off value of ß-hCG in predicting ongoing pregnancy was 147 IU/L (sensitivity 88.3 %, specificity 80.7 %). For vitrified embryo transfers, the value was 135 IU/L (sensitivity 84.4 %, specificity 76.3 %). CONCLUSIONS: Day 12 ß-hCG levels after FET are significantly affected by the methods of embryo cryopreservation for ongoing pregnancies. Furthermore, when using ß-hCG cut-off value to assess pregnancy outcome, the cryopreservation methods should be taken into account.

The effect of progesterone level prior to oocyte retrieval on the numbers of oocytes retrieved and embryo quality in IVF treatment cycles: an analysis of 2,978 cycles.

PURPOSE: The study was designed to evaluate the relationship between serum progesterone (P4) response after hCG administration and the number of oocytes retrieved and the embryo quality in fresh IVF cycles. METHODS: We conducted a retrospective cohort study of women aged 24-43 years who underwent first fresh IVF cycle from 2011 to 2013 at a single practice. We compared the post-hCG serum P4 level with values on the day of hCG trigger. Patients were analyzed in long and short protocols independently. In addition, patients were stratified by post-hCG P4 response. Number of oocytes retrieved and embryo quality were the primary outcomes of interest. Ordinary least square regression models and logistic regression analysis models were created to identify predictive factors associated with embryological outcomes while adjusting for potential confounders. RESULTS: Among the 2,978 IVF cycles, 2,484 patients were in long protocols, and 494 patients were in short protocols. After adjusting for patient age, rFSH duration, and basal FSH levels, the associations between P4 response after hCG administration and number of oocytes retrieved (P < 0.001) remained statistically significant in both long and short protocols. Additionally, mature oocyte rate, fertilization rate, good quality embryo rate, pregnancy rate and implantation rate were not significantly associated with the P4 increase when adjusting for the same factors. However, pregnancy rate and implantation rate from frozen-thawed cycles increased gradually across the seven groups. CONCLUSIONS: Post-hCG P4 levels were positively associated with the number of oocytes retrieved, but did not affect oocyte or embryo quality. Our study suggests that the change in the post-hCG P4 level is another parameter that can be used by clinicians to assess the number of oocytes retrieved, and may further to estimate the pregnancy rate and live birth rate indirectly.

Effect of cumulus cell removal 4 h post-insemination on fertilization and embryo quality: a prospective randomized sibling-oocyte study.

PURPOSE: The study was designed to evaluate whether cumulus cell removal 4 h post-insemination could influence fertilization and embryo quality. METHODS: The study included 61couples undergoing standard long down regulation protocol from July 2011 to May 2012. Sibling oocytes of each patient were randomly assigned to either the 4 h group or the 20 group. For the 4 h group, cumulus cells were removed 4 h after gamete coincubation; for the 20 group, cumulus cells removal was performed 20 h after insemination. Fertilization rate, embryo quality, pregnancy rate and implantation rate were assessed. RESULTS: A total of 801 sibling cumulus-oocyte complexes (COCs) were randomized to the 4 h group (421 COCs) or 20 h group (380 COCs). There was no difference in the two pronuclei, one pronucleus and grade 1-2 embryo rate. Three pronuclei rate was significantly higher in the 4 h group compared to the 20 h group (12.6 % vs. 8.2 %, P = 0.041). Comparison of embryo transfer cycles in which either embryos from the 4 h group or 20 h group were transferred did not reveal any statistically significant differences in pregnancy or implantation rates. CONCLUSION: The results of the present study indicate that cumulus cell removal 4 h post-insemination may increase the percentage of tripronuclear zygotes. However, normal fertilization rate, embryo development, clinical pregnancy rate and implantation rates are not influenced by the timing of cumulus cell removal.

Clinical outcomes after elective double-embryo transfer in frozen cycles for women of advanced maternal age: A retrospective cohort study.

ABSTRACT: We aimed to determine the clinical outcome of double cleavage-stage embryo transfers in frozen-thawed embryo transfer cycles for older women.This study analyzed a total of 8189 cleavage-stage frozen-thawed embryo transfer cycles between January 2013 and December 2017 at Sir Run Run Shaw Hospital. All cycles were sorted into 3 groups based on patient age: ≤35 years (Group A), 36 to 37 years (Group B), and ≥38 years (Group C). The clinical pregnancy rate (CPR), implantation rate (IR), live birth rate (LBR), miscarriage rate, multiple pregnancy rate (MPR), preterm birth rate, and low-birth-weight rate were compared between the 3 groups.Significant differences in CPR, IR, LBR, MPR, and premature birth rate were found among the 3 groups. The CPR, IR, LBR, and MPR in Group A were higher than those in Group C. Transfers of 2 high-quality embryos resulted in significant differences in CPR, IR, LBR, MPR, and neonatal weight among the 3 groups, but no differences in premature birth and abortion rates were observed. Transfers with 1 high-quality and 1 fair-quality embryo resulted in significant differences in CPR, IR, and LBR among the 3 groups. Comparison of transfers of 2 high-quality embryos with 1 high-quality and 1 fair-quality embryo showed that the CPR and LBR were significantly lower for the latter in Groups A and C, but Group B had no salient changes.Higher IR and LBR and lower MPR may be achieved by selection of optimal embryo types for patients of different ages. Two high-quality embryos need to be transferred in women older than 38 years. For women aged 36 to 37 years, 1 high-quality embryo or 1 high-quality plus 1 fair-quality embryo should be singled out for transfer. For women younger than 35 years, a single high-quality embryo should be selected for transfer.

Are testicular sperms superior to ejaculated sperms in couples with previous ART failure due to high rate of fragmented embryos? A retrospective cohort study.

Objective: The aim was to clarify whether using testicular sperm reduces embryo fragmentation and improves cycle outcomes. Methods: Fragmented embryo was defined as an embryo in which fragments account for more than one third of the embryonic surface area. High rate of fragmented embryos was defined by a proportion of fragmented embryos higher than 50%. We recruited infertile couples who had undergone at least one ovarian stimulation cycle using ejaculated sperm but failed to conceive due to high rate of fragmented embryos in each previous cycle. After fully informed consent, the couples agreed to obtain testicular sperm by testicular puncture and use testicular sperm for intracytoplasmic sperm injection (ICSI). The normal fertilization rate, transferable embryo rate, fragmented embryo rate and cycle outcomes were compared between ejaculated sperm group (EJA-sperm group) and testicular sperm group (TESTI-sperm group). Results: Twenty-two couples who agreed to participate in our study underwent 32 ICSI cycles with ejaculated spermatozoa and 23 ICSI cycles with testicular spermatozoa. Embryo transfers were cancelled in 8 ejaculated cycles and 4 testicular cycles because of no transferable embryos. There were no significant differences in age, normal fertilization rate and high-quality embryo rate between ejaculated and testicular groups. The transferable embryo rate and implantation rate in TESTI-sperm group were significantly higher than those in EJA-sperm group (36.9% vs. 22.0%, p < 0.01; 34.2% vs. 0%, p < 0.001). The fragmented embryo rate in TESTI-sperm group was significantly lower than that in EJA-sperm group (61.2% vs. 75.7%, p < 0.05). Conclusion: Our small retrospective cohort study suggests that using testicular sperm may be a recommended option for couples with previous ART failure because of high rate of fragmented embryos. Large samples, multicenter studies or randomized controlled trial (RCT) are needed to further confirm the superiority of testicular sperm.

[Analysis of low-quality embryo transfer strategy in in vitro fertilization and embryo transfer cycle].

OBJECTIVE: To investigate the transfer strategy of low-quality embryo in in vitro fertilization and embryo transfer (IVF-ET) cycle. METHODS: A retrospective analysis was performed for the clinical data of 621 IVF-ET cycles under controlled ovarian hyperstimulation, including 574 fresh embryo transfer (ET) cycles (Group T1) and 47 frozen-thawed embryo transfer (FET) as the first ET cycles (Group C1). Logistic regression was used to model the probability of clinical pregnancy rate based on the cycle-specific factors. RESULTS: The clinical pregnancy rate was significantly higher in Group C1 than Group T1 [38.3% (18/47) vs 22.1% (127/574), P < 0.05]. Multivariate logistic regression analysis revealed that patient age and ET method were significantly associated with the clinical pregnancy rate. After adjusting for patient age, the clinical pregnancy rate remained significantly higher in Group C1 than Group T1 (OR: 2.107, 95%CI: 1.128 - 3.939, P < 0.05). CONCLUSION: The use of FET instead of fresh ET may improve the clinical pregnancy rate in low-quality embryo cycles.

[Risk factors associated with twin pregnancy in double embryo transfer].

OBJECTIVE: To explore the risk factors associated with twin pregnancy in double embryo transfer. METHODS: A retrospective analysis was performed for 2970 double embryo transfer cycles, including 1984 cycles of fresh embryo transfer and 986 cycles of frozen-thawed embryo transfer (FET). Multiple factor Logistic regression was used. The twin pregnancy rate was studied in group of age < 35 or age ≥ 35 years old, in group of zero, single or double high-quality embryos transferred and group of fresh embryo transfer or FET cycles. RESULTS: (1) There was a significantly negative correlation between women's age and twin pregnancy according to the analysis of multiple factor Logistic regression (P < 0.01). And a significantly positive correlation existed between the number of high-quality embryos transferred, FET and twin pregnancy (P < 0.01); (2) the twin pregnancy rate of both fresh embryo transfer and FET cycles was higher in group of age < 35 years than that in group of age ≥ 35 years (16.0% vs 8.0%, P < 0.01; 26.9% vs 14.2%, P < 0.01); (3) the twin pregnancy rate of fresh embryo transfer cycles with double high-quality embryos transferred was higher than that with zero or single high-quality embryos transferred (19.1% vs 5.4%; 19.1% vs 11.0%, P < 0.01); the twin pregnancy rate of FET cycles with double high-quality embryos transferred was also higher than that with zero or single high-quality embryos transferred (32.7% vs 10.8%; 32.7% vs 20.7%, P < 0.01); (4) the twin pregnancy rate of FET cycles was significantly higher than that of fresh embryo transfer cycles (24.7% vs 14.9%, P < 0.01). CONCLUSION: Women's age, the number of high quality embryos transferred and FET are risk factors associated with twin pregnancy. At reproductive centers with a mature technique of FET, we recommend performing single high-quality embryo transfer in FET cycles to reduce the occurrence of twin pregnancy.

CFTR is essential for sperm fertilizing capacity and is correlated with sperm quality in humans.

BACKGROUND: Our previous studies have demonstrated the cystic fibrosis transmembrane conductance regulator (CFTR) is important for capacitation and male fertility in mouse and guinea pig spermatozoa. However, the exact function of CFTR on human sperm fertilizing capacity, and correlation with sperm quality has not been established. The present study may shed light on some unexplained male infertility, and on a possible new method for diagnosis of male infertility and strategy for male contraception. METHODS: To assess the effect of CFTR on human sperm fertilizing capacity, we examined sperm capacitation and the acrosome reaction using chlortetracycline staining, analyzed sperm hyperactivation by computer-assisted semen analysis (CASA), measured intracellular cAMP levels using ElA and evaluated sperm penetration of zona-free hamster eggs assay in fertile men. The percentage of spermatozoa expressing CFTR from fertile, healthy and infertile men (mainly teratospermic, asthenoteratospermic, asthenospermic and oligospermic) was conducted by indirect immunofluorescence staining. RESULTS: Progesterone significantly facilitated human sperm capacitation and ZP3 triggered the acrosome reaction, both were significantly inhibited by CFTR inhibitor-172 (CFTRinh-172; 10 nM-1 microM) in a dose-dependent manner. The presence of 100 nM CFTRinh-172 markedly depressed intracellular cAMP levels, sperm hyperactivation and sperm penetration of zona-free hamster eggs. In addition, the percentage of spermatozoa expressing CFTR in the fertile men was significantly higher than healthy and infertile men categories (P < 0.01). CONCLUSIONS: CFTR is essential for human sperm fertilizing capacity and the impairment of CFTR expression in spermatozoa is correlated with a reduction of sperm quality. These results suggest that defective expression of CFTR in human sperm may lead to the reduction of sperm fertilizing capacity.

[Fertilizing ability, cleavage potential and inheritance risk of globozoospermia].

OBJECTIVE: To explore the fertilizing ability, cleavage potential and inheritance risks of globozoospermia. METHODS: A globozoospermic patient was diagnosed by sperm morphological staining and transmission electron microscope. From his wife the investigators obtained 26 oocytes in which 6 oocytes were donated and the other 19 (in 20) MII oocytes injected into 19 round-headed sperms. Six donated oocytes accepted in vitro fertilization (IVF). This patient's chromosome and microdeletion in AZFa, AZFb and AZFc in Y chromosome were checked through 6 sequence tag sites of sY84, sY86, sY127, sY134, sY254 and sY255. RESULTS: Only 4 (in 19) were normally fertilized and cleaved. All 6 donated oocytes were normally fertilized and cleaved. The fertilizing rate was significantly higher than that of this patient (100% vs 21.1%, P < 0.01). But the cleavage rate has no statistical difference (100% vs 100%, P > 0.05). This patient had normal chromosome (46, XY) and there was no deletion in Y chromosome. His wife became pregnant after accepting two thawed embryos and then gave birth to a boy whose chromosome was normal (46, XY). The acceptor also gave birth to a healthy boy after accepting two thawed embryos. CONCLUSIONS: Despite a lower fertilizing rate, intracytoplasmic sperm injection is still an effective therapy for globozoospermic infertility.

CFTR gene mutations and polymorphism are associated with non-obstructive azoospermia: From case-control study.

A variety of experimental studies have yielded evidence that the cystic fibrosis transmembrane conductance regulator (CFTR) protein participates in the process of spermatogenesis. However, the association between CFTR gene and non-obstructive azoospermia (NOA) disease remained to be a question. First, we reviewed available data from the PubMed and Embase databases before May 2016 to find the most common mutations of CFTR gene in NOA patients. Second, an original case-control study was conducted on NOA patients (n=100) and a control group consisting of fertile males (n=100), selected from August 2015 to March 2017, to detect CFTR gene mutations and polymorphism. Peripheral blood samples from NOA patients and normal controls were analyzed for the presence of specific sequences of CFTR gene by polymerase chain reaction amplification followed by direct sequencing. From our comprehensive review, 12 case-control studies were found concerning the relation between CFTR gene mutations and polymorphism and NOA disease. Fifty-four mutations were mentioned and IVS8 poly-T, TG repeats, F508del and R117H mutations were the most common ones. Based on that, we detected IVS8 poly-T, TG repeats, F508del, R117H and M470V mutations in our case control study. We found that the T5 allele was present at a significantly higher rate in NOA patients than in the control group (5.00% versus 0.00%, p<0.01) with increased risk having NOA [Odds ratios (OR) 2.05, 95% confidence intervals (CI) 1.85-2.27]. The T5 variant was always accompanied by TG12 (10/10) and V470 allele participated in most TG12T5 haplotypes (8/10). TG12T5-V470 haplotype also enhanced risk of having NOA [OR 2.04, 95% CI 1.84-2.26]. F508del and R117H mutations were not found in either group. In conclusion, the polyvariant mutant genes of CFTR: T5 allele and TG12-T5-V470 genotype are correlated with NOA, but F508del and R117H mutations have low possibility to be associated with NOA.

[Comparison between the results of ICSI with fresh and with frozen-thawed sperm obtained by PESA to treat azoospermia].

OBJECTIVE: Retrospective study of the results of ICSI (intracytoplasmic sperm insemination) with frozen sperm obtained by PESA (percutaneous epididymal sperm aspiration) was performed in 27 patients. METHODS: With conventional freezing method, sperm from diagnosing PESA and the remaining motile sperm after treating cycle were frozen. After frozen-thawed and ICSI process, fertilization rate, implantation rate, clinical pregnancy rate were compared and other outcomes including pregnant combinations and parameters of newborns of experimental group (which used frozen-thawed sperm) and control group (which used fresh PESA sperm) were analyzed respectively. RESULTS: One hundred and sixty three and 1 157 oocytes of stage M II were injected respectively in the experimental group (15 cycles) and control group (100 cycles), and fertilization rate of experimental group was prominently higher than that of control group (84.05% vs 73.29%, P < 0.05), while implantation rate and clinical pregnancy rate were of no difference from the control, respectively (23.07% vs 15.73%; 53.33% vs 37.00%, P > 0.05). The differences in newborn's weights between two groups were of no statistical significance (P > 0.05). In the experimental group, eight clinical pregnancies were achieved including 5 live deliveries and 3 ongoing pregnancies, 37 clinical pregnancies including 30 deliveries with only 1 fetal death, 3 ongoing pregnancies and 4 abortions in the control group. Neither vital pregnant combinations nor neonate malformations were found in both groups. CONCLUSION: ICSI using frozen-thawed sperm obtained by PESA is an economic effective and safe method to treat azoospermia. Recovering rates of frozen sperm form PESA should be further increased.

Characteristics of nonylphenol and bisphenol A accumulation by fish and implications for ecological and human health.

Fish populations constitute an important part of aquatic ecosystems. Thus, their accumulation of nonylphenol (NP) and bisphenol A (BPA) may pose risks to ecosystems and human health. This study analyzed the concentrations of NP and BPA in four types of fishes (i.e., wild/farmed freshwater fishes and wild/farmed marine fishes). Wild freshwater fishes contained higher concentrations of NP and BPA than the other three types of fishes. The concentrations of NP in the wild freshwater fishes ranged from 1.01 to 277 µg/kg ww, with bioconcentration factors (BCFs) and biota-sediment accumulation factors (BSAFs) ranging from 74.0 to 2.60 × 10(4)L/kg and from 0.003 to 18.3, respectively. The wild freshwater fishes contained relatively low amounts of BPA, varying from ND to 25.2 µg/kg ww, with the BCFs and BSAFs ranging from 1.00 to 274L/kg and from 0.003 to 3.40, respectively. Five fish species particularly showed high BCFs and BSAFs, indicating that they could be an important source of NP for higher trophic levels, most likely resulting in ecological risks. The demersal fishes showed a greater ability to accumulate NP than the pelagic ones. The fact that the 95th percentile values of the risk quotient (RQ) for NP and BPA were higher than the acceptable threshold indicated that these two compounds would have adverse effects on aquatic organisms in Taiwanese rivers. The consumption of wild marine fishes had the highest 95th percentile values of hazard quotient (HQ) for NP and BPA among the four types of fishes, particularly for the population aged 0-3 years. However, the 95th percentile values of HQ for NP and BPA were all less than 1, suggesting that exposure to NP and BPA through fish consumption posed no remarkable risk to human health in Taiwan.