Clinical and neonatal outcomes of using a modified micro cryotube for cryopreservation of small numbers of spermatozoa for TESA-ICSI cycles.

Cryopreservation of a small number of human spermatozoa is still a major challenge for embryologists. The aim of this study was to evaluate the clinical pregnancy and neonatal outcomes of intracytoplasmic sperm injection (ICSI) using a modified micro cryotube as freezing carrier for freezing small numbers of human spermatozoa collected by testicular sperm aspiration (TESA). We conducted a retrospective study to analyses the ICSI outcomes of using frozen-thawed few testicular spermatozoa in males with obstructive azoospermia (OA) from June 2017 to June 2021. Of 155 ICSI treatment cycles, 79 cycles were allocated to frozen sperm group and a modified micro cryotube was used for freezing testicular sperm, 76 cycles were allocated as fresh sperm group. No significant differences were observed in fertilization rate, good quality embryo rate, and blastocyst rate between the frozen sperm group and fresh sperm group (P > 0.05). Similarly, in the fresh embryo transfer cycles plus the first frozen-thawed embryo transfer cycles, the total clinical pregnancy rate (54.43% vs 57.89%), implantation rate (46.08% vs 49.47%), miscarriage rate (13.95% vs 13.64%) and live birth rate (45.57% vs 48.68%) were not statistically different between the frozen and fresh sperm groups (P > 0.05). In addition, there was no statistical differences in the mean gestational age (38.33weeks ± 1.74 vs 37.89weeks ± 1.87), preterm delivery rate (5.56% vs 10.81%), mean birth weight at delivery (3026.50 g ± 577.64 vs 2977.56 g ± 528.93), and low birth weight (12.50% vs 19.51%) between the two groups (P > 0.05 in all cases). Modified micro cryotube for cryopreservation of rare testicula rretrieved spermatozoa did not negatively affect the pregnancy and neonatal outcomes in TESA-ICSI cycles. The presented method may be a useful alternative for cryopreservation of small numbers of human spermatozoa in clinical setting.

Recurrent Implantation Failure May Be Identified by a Combination of Diagnostic Biomarkers: An Analysis of Peripheral Blood Lymphocyte Subsets.

Background: Recurrent implantation failure (RIF) is a challenge during assisted reproductive technology (ART). In the present study, potential diagnostic biomarkers for the immune status of peripheral blood lymphocyte subsets in patients with RIF were analyzed, with the aim of identifying novel biomarkers that may predict RIF. Methods: A total of 41 participants, including 21 women with RIF and 20 fertile controls, were included in the present study. Functional analysis was performed and the cytokine status of natural killer (NK), T, CD8+ T, T helper (Th), and γδ T cells which are lymphocyte subsets in peripheral blood was measured using flow cytometry. Binary logistic regression analysis adjusted for T follicular helper 1 (Tfh1), Tfh2, Tfh17, and early NK cells was performed to determine the relationship between the peripheral blood lymphocyte subsets and RIF. Potential diagnostic biomarkers were assessed by logistic regression analysis and receiver operating characteristic curves. Results: There were significantly more Tfh1, Tfh17, and NK cells in the RIF group compared with the control group (all P < 0.05). However, the percentage of T, regulatory T (Tregs), and Tfh2 cells, as well as early inhibitory NK cells, was significantly lower in the RIF group compared with the control group (all P < 0.05). Following logistics regression analysis, Treg, Tfh17, and early inhibitory NK cells exhibited significant differences between the two groups. Combination diagnosis using these 3 biomarkers had a higher area under the curve of 0.900 (95% confidence interval: 0.808-0.992, P < 0.001) in the RIF group compared with that in the control group. Conclusion: T, Tregs, Tfh1, Tfh2, Tfh17, NK cells, and early inhibitory NK cells may play important regulatory roles in embryo implantation. The combination of 3 molecular markers (Treg, Tfh17, and early inhibitory NK cells) could provide a high diagnostic value for women with RIF, thus providing novel potential biomarkers for RIF in ART. The present findings could provide a reference either for the clinical treatment of patients with RIF or for future large, well-designed studies.

Laser-assisted selection of immotile spermatozoa has no effect on obstetric and neonatal outcomes of TESA-ICSI pregnancies.

BACKGROUND: Azoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable, immotile spermatozoa before the procedure. There are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI. METHODS: A retrospective comparative study was conducted on outcomes of ICSI cycles with testicular spermatozoa from June 2014 to June 2018. Of 132 cycles, 33 were allocated to the test group and oocytes were injected with immotile spermatozoa selected by laser, 99 cycles were allocated as control group. RESULTS: Compared with the control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the test group in either fresh or frozen transfer cycles. The cumulative live birth rate in the test group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight, and the malformation rate between the test and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05). CONCLUSIONS: No negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.

Choosing the optimal blastocyst by morphology score versus developmental rate in frozen-thawed embryo transfer cycles.

The aim of this study was to provide guidance for better management in the selection of blastocyst to warm in frozen-thawed embryo transfer (FET) cycles. A retrospective cohort follow-up study was conducted that included single autologous frozen blastocyst transfer cycles performed in our Reproductive Medicine Unit from January 2009 to December 2016. The live birth rate (LBR), clinical pregnancy rate (cPR) were increased as blastocyst morphology scores increased, but the miscarriage rate decreased in all groups. In the high-score groups, there were no differences in LBR between D5 and D6, while in the low-score groups, LBR was significantly higher in D5 compared to the D6. With respect to neonatal outcome, there were no differences in all the groups. After binary logistic regression analysis, it was seen that patients' age, thawed cycles, pre-frozen morphology score and developmental rate were independently associated with LBR. These results suggest that for high-scoring blastocyst, the pre-frozen morphological score should be given priority while for low-scoring blastocysts, the developmental rate should be given priority when thawing in FET cycles.

Comparable clinical outcomes and live births after single vitrified-warmed and fresh blastocyst transfer.

Selective single-blastocyst transfer (SBT) in fresh cycles has been effective in reducing multiple pregnancies. However, we do not know whether this successful strategy of fresh transfer cycles is suitable for cryopreserved cycles. The present study was undertaken to evaluate the feasibility and value of SBT in vitrified-warmed cycles. Clinical pregnancy rate (CPR) was similar with vitrified and fresh SBT (46.61% versus 52.15% respectively). Of the pregnant patients, monozygotic twin, miscarriage and ectopic pregnancy rates were similar with vitrified and fresh SBT. For the newborns, no significant difference was observed in live birth, low birthweight, premature delivery and birth defects rates between vitrified and fresh SBT. With respect to the quality of transferred blastocysts (from BB to AA), a similar CPR and miscarriage rate was obtained for both vitrified and fresh SBT when a similar blastocyst cohort graded ≥ 3BB was transferred. The data show that vitrified SBT is an effective means of reducing multiple pregnancy and that comparable clinical outcomes and live births are achieved if single blastocysts graded ≥ 3BB are transferred for both vitrified and fresh SBT. These data should encourage clinics to evaluate their embryo transfer policy and adopt vitrified SBT as everyday practice. Selective single-blastocyst transfer in fresh cycles has been an effective method to reduce the multiple pregnancies. However, due to a lack of adequate studies, we do not know whether this successful strategy in fresh transfer cycles is suitable in cryopreserved cycles. The present study was undertaken to explore the feasibility and value of single-blastocyst transfer in vitrified-warmed cycles. We found that single-blastocyst transfer in vitrified-warmed cycles is an effective means of reducing multiple pregnancy, and comparable clinical outcomes and live births were achieved if single blastocysts graded ≥ 3BB were transferred for both vitrified-warmed and fresh blastocyst transfer. These data should encourage clinics to evaluate their embryo transfer policy and adopt single-blastocyst transfer in cryopreserved cycles as their everyday practice.

[The nasal development after one-staged correction of nose deformity and unilateral complete cleft lip in infancy].

OBJECTIVE: To introduce one-staged correction of nasal deformity and unilateral complete cleft lip in infancy and to observe the nasal development after the operation. METHODS: The unilateral complete cleft lip and nasal deformity were corrected in one stage in27 cases. They were followed up for several years. With post-operative photos, the anthropometric method was used to analyze the nasal development. RESULTS: The long-term results were excellent in 10 cases, good in 14 cases, and poor in 3 cases. CONCLUSIONS: Based on the anatomic findings of nasal blood supply, one-staged correction of nasal deformity and unilateral complete cleft lip in infancy can be performed with no obvious interference with nasal development. The secondary nasal deformity before school age can be alleviated or avoided.

[A clinical study of presurgical nasoalveolar molding in infants with complete cleft lip and palate].

OBJECTIVE: To observe the clinical effect of presurgical nasoalveolar molding in infants with complete cleft lip and palate. METHODS: Presurgical nasoalveolar molding was performed in 38 infants with cleft lip and palate (26 patients with unilateral cleft lip and palate, 12 patients with bilateral cleft lip and palate), aged between 5 and 30 days. The width of alveolar cleft was measured before and after the operation and the effect of treatment was assessed. RESULTS: After 108 - 152 days of therapy, the average width of alveolar cleft decreased by 5.3 mm in 26 patients with unilateral cleft lip and palate. Nasal profile was improved in 76 percent of cases. In 12 patients with bilateral cleft lip and palate, the average width of left cleft decreased by 4.7 mm and that of the right decreased by 4.2 mm. The distance between right and left cleft increased by 5.1 mm. Nasal profile was improved in 66 percent of cases. CONCLUSION: Presurgical nasoalveolar molding in complete cleft lip and palate can improve nasal profile and decrease the width of alveolar cleft.

[Simultaneous repair of complete cleft lip and palatein infancy-preliminary observation (271 cases report)].

OBJECTIVE: To Verify the safety and reliability of one-stage repair of complete cleft Lip and palate in infancy and to obtain the primary result. METHODS: The simultaneous repair of complete cleft Lip and palate in infants 3 to 12 months of age were performed in 271 cases. The deformities include 185 cases of typical complete unilateral clefts and 75 cases of complete bilateral clefts, and other 11 atypical cleft infants. The preoperative orthopedic treatment for wide alveolar cleft was undertaken in 24 infants and the lip appearance and speech outcome were evaluated in 116 children by 1 to 4 years' postoperative follow-up. RESULTS: All infants, except for dyspnea in 2 babies, palatal fistula formation in 6 cases and temporary wound hemorrhage in 5 infants, were recovered without complications. After orthopedic treatment, the width of the alveolar cleft was reduced 6.1 mm in average. The evaluation showed that 93.1% of children had got good or excellent lip appearance. And the acceptable or excellent speech was found in 94.8% children. CONCLUSIONS: Simultaneous repair of complete cleft lip and palate in infancy is safety and reliable. The preoperative orthopedic procedure is able to reduce the wide alveolar cleft and to achieve alignment of alveolar segments. The acceptable and or excellent lip appearance and speech function could be obtained in this one-stage operative procedure in infants.