The impact of oocytes containing smooth endoplasmic reticulum aggregates on assisted reproductive outcomes: a cohort study.

BACKGROUND: The impact of smooth endoplasmic reticulum aggregates (SERa) on assisted reproductive technology (ART) outcomes was still controversial. Our objective is to investigate the impact of the presence of SERa on intracytoplasmic sperm injection (ICSI) outcomes. METHODS: This was a retrospective cohort study. A total of 1,090 fresh ICSI cycles from 944 patients between January 2016 and June 2020 were included. Outcomes from clinical, embryological and neonatal aspects were compared between SERa + and SERa- cycles as well as between SERa + and SERa- oocytes. RESULTS: The total gonadotropin (Gn) dose, number of oocytes retrieved, serum estradiol concentration and number of the available embryo were significantly higher in SERa + cycles than in SERa- cycles (P < 0.05). Comparable two pronuclei (2PN) fertilization rate and poly-pronucleus zygote rate were shown in SERa + and SERa- cycles (P > 0.05), but which were higher in SERa + oocytes than in SERa- oocytes (P < 0.05). No statistical difference in blastocyst formation rate was found in SERa + and SERa- cycles as well as in SERa + and SERa- oocytes (P > 0.05). Good-quality embryo rate was statistically higher in SERa- cycles than in SERa + cycles (P < 0.05), but the difference was comparable between SERa + and SERa- oocytes (P > 0.05). No statistical difference in clinical pregnancy rate, spontaneous abortion rate, live birth rate and premature delivery rate were found in SERa + and SERa- cycles as well as in SERa + and SERa- oocytes (P > 0.05). The implantation rate was comparable in SERa + and SERa- cycles (P > 0.05), but it is higher in the group of only SERa- embryo transfer when compared with the group of mixed SERa + and SERa- embryo transfer (P < 0.05). 159 newborns in SERa + cycles and 140 newborns in SERa- cycles were followed up. Comparable newborn malformation rate was observed between SERa + and SERa- cycles and oocytes (P > 0.05). Logistic regression analysis revealed number of oocytes and total dose of Gn were risk factors for SERa occurrence (aOR = 1.05 and 1.55, P < 0.001). CONCLUSION: Oocyte's SERa is correlated with a number of oocytes retrieved and higher Gn dose, but it does not affect pregnancy outcomes and increase newborn malformation rate.

Common dysmorphic oocytes and embryos in assisted reproductive technology laboratory in association with gene alternations.

Amorphic or defected oocytes and embryos are commonly observed in assisted reproductive technology (ART) laboratories. It is believed that a proper gene expression at each stage of embryo development contributes to the possibility of a decent-quality embryo leading to successful implantation. Many studies reported that several defects in embryo morphology are associated with gene expressions during in vitro fertilization (IVF) treatment. There is lacking literature review on summarizing common morphological defects about gene alternations. In this review, we summarized the current literature. We selected 64 genes that have been reported to be involved in embryo morphological abnormalities in animals and humans, 30 of which were identified in humans and might be the causes of embryonic changes. Five papers focusing on associations of multiple gene expressions and embryo abnormalities using RNA transcriptomes were also included during the search. We have also reviewed our time-lapse image database with over 3000 oocytes/embryos to show morphological defects possibly related to gene alternations reported previously in the literature. This holistic review can better understand the associations between gene alternations and morphological changes. It is also beneficial to select important biomarkers with strong evidence in IVF practice and reveal their potential application in embryo selection. Also, identifying genes may help patients with genetic disorders avoid unnecessary treatments by providing preimplantation genetic testing for monogenic/single gene defects (PGT-M), reduce embryo replacements by less potential, and help scientists develop new methods for oocyte/embryo research in the near future.

Pregnancy outcomes in patients with polycystic ovary syndrome who conceived after single thawed blastocyst transfer: a propensity score-matched study.

BACKGROUND: It remains unclear whether polycystic ovary syndrome (PCOS) is an independent risk factor for pregnancy complications in women undergoing assisted reproductive technology (ART) treatment. For the integrative treatment of PCOS patients, it is still important to investigate the pregnancy outcomes of PCOS patients after adjusting for potential biases, such as body mass index, embryo quality and endometrial preparation method. METHODS: This retrospective cohort study ultimately included a total of 336 PCOS patients who conceived after single thawed blastocyst transfer in the PCOS group and 2,325 patients in the control group from January 2018 to December 2020. A propensity score matching (PSM) model was used, and 336 PCOS patients were matched with 336 patients in the control group. RESULTS: Before PSM, no differences in the miscarriage rate, pregnancy complication rate, preterm birth rate, or live birth rate were found between the PCOS group and the control group. After PSM, the late miscarriage rate of the PCOS group was significantly higher than that of the control group (3.3% vs. 0.6%, P = 0.040), although the early miscarriage rates were similar (14.0% vs. 13.7%). The rates of pregnancy complications, preterm birth and live birth in the PCOS group were comparable to those in the matched control group (P = 0.080, P = 0.105, P = 0.109, respectively). The neonatal weights of male infants and female infants were similar between the two groups (P = 0.219, P = 0.169). Subgroup analysis showed that PCOS patients with homeostasis model assessment of insulin resistance (HOMA-IR) levels ≥ 2.49 had a significantly increased risk of preterm birth compared with those with HOMA-IR levels < 1.26 and 1.26 ≤ HOMA-IR levels < 2.49 (26.0% vs. 6.0% vs. 9.8%, P = 0.005). PCOS patients with total testosterone levels ≥ 0.7 ng/ml had a higher early miscarriage rate but a lower late miscarriage rate than those with total testosterone levels < 0.7 ng/ml (29.4% vs. 12.3%, 0% vs. 3.6%, respectively, P = 0.032). CONCLUSIONS: PCOS is an independent risk factor for late miscarriage in patients conceived after a single thawed blastocyst transfer, even after adjusting for biases. Among PCOS patients, insulin resistance and hyperandrogenism are associated with a higher risk of preterm birth and early miscarriage, respectively.

Development and Validation of a Novel Prognostic Nomogram Combined With Desmoplastic Reaction for Synchronous Colorectal Peritoneal Metastasis.

Purpose: The prognostic value of desmoplastic reaction (DR) has not been investigated in colorectal cancer (CRC) patients with synchronous peritoneal metastasis (SPM). The present study aimed to identify whether DR can predict overall survival (OS) and develop a novel prognostic nomogram. Methods: CRC patients with SPM were enrolled from a single center between July 2007 and July 2019. DR patterns in primary tumors were classified as mature, intermediate, or immature according to the existence and absence of keloid-like collagen or myxoid stroma. Cox regression analysis was used to identify independent factors associated with OS and a nomogram was developed subsequently. Results: One hundred ninety-eight and 99 patients were randomly allocated into the training and validation groups. The median OS in the training group was 36, 25, and 12 months in mature, intermediate, and immature DR categories, respectively. Age, T stage, extraperitoneal metastasis, differentiation, cytoreductive surgery (CRS), hyperthermic intraperitoneal chemotherapy (HIPEC), and DR categorization were independent variables for OS, based on which the nomogram was developed. The C-index of the nomogram in the training and validation groups was 0.773 (95% CI 0.734-0.812) and 0.767 (95% CI 0.708-0.826). The calibration plots showed satisfactory agreement between the actual outcome and nomogram-predicted OS probabilities in the training and validation cohorts. Conclusions: DR classification in the primary tumor is a potential prognostic index for CRC patients with SPM. The novel prognostic nomogram combined with DR classification has good discrimination and accuracy in predicting the OS for CRC patients with SPM.

Effects of nonoxynol-9 (N-9) on sperm functions: systematic review and meta-analysis.

Objective: To summarize the currently available phase I and II clinical trials of the effects of nonoxynol-9 (N-9) on human sperm structure and functions. Methods: A systematic review and meta-analysis aiming to evaluate the spermicidal activity of N-9 on motility, was conducted in PubMed, EMBASE, and Cochrane databases by 10 March 2021. The counted numbers of progressive motile (PR) sperm in cervical mucus and the vanguard sperm penetration distances were analyzed. Other effects on sperm structures and physiological activities were reviewed as well. Results: In the pooled results, percentages or counted numbers of PR sperm decreased after the treatment of N-9. Vanguard sperm penetration distance was shortened in treated groups. N-9 has been confirmed to damage the structures of sperm, as well as other organelles like acrosome and mitochondria. The physiological activities such as generation of reactive oxygen species, superoxide dismutase activity, acrosin activity, and hemizona binding were all inhibited in the reviewed studies. Conclusions: N-9 has several impacts on sperm owing to its potency in reducing sperm motility and cervical mucus penetration, as well as other functional competencies. Lay summary: Nonoxynol-9 (N-9) has been used worldwide as a spermicide to kill sperm for more than 60 years but can cause side effects including vaginal irritation and can increase the rate of contraceptive failure. A detailed analysis of published literature aiming to evaluate the spermicidal activity of N-9 on sperm was carried out. In the pooled results, N-9 reduced the number of active sperm and the distance they traveled. It also caused damage to the structures of sperm and to the way the sperm acted and interacted with the egg. In conclusion, N-9 impacts on sperm in a number of ways that lead to sperm death and dysfunction.

Associations between ambient air pollution and IVF outcomes in a heavily polluted city in China.

RESEARCH QUESTION: Is air pollution related to IVF outcomes in a heavily polluted city in China? DESIGN: A retrospective cohort study of 8628 fresh, autologous IVF cycles was conducted for the first time at the Reproductive Medicine Center of The Third Affiliated Hospital of Zhengzhou University between May 2014 and December 2018 (oocyte retrieval date). The exposure was divided into four periods (gonadotrophin injection to oocyte retrieval [P1], oocyte retrieval to embryo transfer [P2], 1 day after embryo transfer to embryo transfer +14 days [P3] and gonadotrophin injection to embryo transfer +14 days [P4]) and four levels (Q1-Q4 according to their 25th, 50th and 75th percentiles). RESULTS: An interquartile range increase (Q2 versus Q1) in particulate matter ≤10 µm (PM10) during P3 and P4 and sulphur dioxide (SO2) during P3 significantly decreased the clinical pregnancy rate (adjusted odds ratio [aOR] 0.81, 95% confidence interval [CI] 0.71-0.92 for PM10 of P3; aOR 0.87, 95% CI 0.76-1.00 for PM10 of P4; aOR 0.82, 95% CI 0.73-0.93 for SO2 of P3). In addition, PM10 was associated with an increased biochemical pregnancy rate (Q3 versus Q1: aOR 1.55, 95% CI 1.09-2.19 for PM10 of P1) and decreased live birth rate (Q2 versus Q1: aOR 0.88, 95% CI 0.77-0.99 for PM10 of P3). The multivariate regression results were consistent with that of multiple treatments propensity score method (PSM) for SO2 pollutants in P3 and PM10 pollutants in P4. CONCLUSION: From the early follicular stage to the pregnancy test period, high concentrations of PM10 and SO2 may have a negative impact on IVF treatment outcomes in the study area.

In vitro rescue immature oocytes - a literature review.

Immature oocytes retrieved from in vitro fertilization (IVF) and clinical in vitro maturation (IVM) is a common problem, especially in patients with advanced age, poor ovarian response (POR), or polycystic ovary syndrome (PCOS). Considering there is no common name to describe this group of oocytes, we suggest naming all of immature oocytes retrieved from IVF and clinical IVM cycles as 'Medical Unusable Oocytes' (MUO) as none of them will be used for subsequent treatment and will eventually be discarded. Scientists attempt to improve the clinical utilization rate of MUO instead of discarding them. Rescue IVM and mitochondria supplementation may be available approaches to mature MUO. We propose a specific definition of rescue IVM, namely the cultivation and maturation of immature oocytes in vitro collected from IVF cycles with human chorionic gonadotropin (hCG) trigger. Rescue IVM is usually mixed up with clinical IVM. Clarification of the differences between rescue IVM and clinical IVM is necessary. This manuscript aims to clarify the rather confusing IVM procedures and review existing methods of improving rescue IVM, currently available information on the success rate, and explore the future possibility of rescue IVM serving as a promising tool in reproductive medicine.

Kidney Biopsy in Patients With Monoclonal Gammopathy: A Multicenter Retrospective Cohort Study.

Objectives: To analyze the clinical characteristics and renal pathological manifestations of patients with monoclonal gammopathy (MG) and kidney injury. Methods: This was a multicenter retrospective cohort study conducted at four tertiary hospitals in China. The study population comprised patients with MG admitted from January 1 2013 to December 31 2020. Hospitalization records, laboratory data, and kidney biopsy reports of all patients were collected from the electronic hospital information systems. The study outcomes included kidney disease progression and major hemorrhagic complications after kidney biopsy. Results: We identified 1,164 patients with MG, 782 (67.2%) of whom had underlying kidney injury. Of 101 patients who underwent kidney biopsy, 16 had malignant neoplasms. Amyloid nephropathy was the most common finding (n = 34, 33.7%), followed by membranous nephropathy (n = 18, 17.8%) and membranoproliferative nephritis (n = 8, 7.9%). Among 85 patients with non-malignant hematologic conditions who underwent kidney biopsy, 43 had MG of renal significance (MGRS) related lesions and 42 had MG-unrelated lesions. The risk of kidney disease progression was higher in patients with kidney injury than in patients without kidney injury. Conclusion: Among patients with MG and kidney injury, only 12.9% underwent kidney biopsy and more than 40% of these patients had MG-unrelated lesions. A kidney biopsy is safe and essential to maximize the possibility of correct diagnosis for patients with clinically suspected MG of renal significance (MGRS).

Application of convolutional neural network on early human embryo segmentation during in vitro fertilization.

Selection of the best quality embryo is the key for a faithful implantation in in vitro fertilization (IVF) practice. However, the process of evaluating numerous images captured by time-lapse imaging (TLI) system is time-consuming and some important features cannot be recognized by naked eyes. Convolutional neural network (CNN) is used in medical imaging yet in IVF. The study aims to apply CNN on day-one human embryo TLI. We first presented CNN algorithm for day-one human embryo segmentation on three distinct features: zona pellucida (ZP), cytoplasm and pronucleus (PN). We tested the CNN performance compared side-by-side with manual labelling by clinical embryologist, then measured the segmented day-one human embryo parameters and compared them with literature reported values. The precisions of segmentation were that cytoplasm over 97%, PN over 84% and ZP around 80%. For the morphometrics data of cytoplasm, ZP and PN, the results were comparable with those reported in literatures, which showed high reproducibility and consistency. The CNN system provides fast and stable analytical outcome to improve work efficiency in IVF setting. To conclude, our CNN system is potential to be applied in practice for day-one human embryo segmentation as a robust tool with high precision, reproducibility and speed.

The predictive value of endometrial thickness in 3117 fresh IVF/ICSI cycles for ectopic pregnancy.

OBJECTIVES: To evaluate the predictive value of endometrial thickness (EMT) during in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles for ectopic pregnancy (EP). METHODS: A total of 3068 patients with 3117 fresh IVF/ICSI cycles between January 2016 and February 2019 from the Reproductive Medicine Center of Sun Yat-Sen Memorial Hospital were included in this retrospective study. The patients were divided into an EP group (n = 92) and an intrauterine pregnancy (IUP) group (n = 3025). Multiple logistic regression analysis was conducted to evaluate the EP risk factors. Receiver operating characteristic (ROC) curves were used to evaluate the predictive value of the risk factors for EP and calculate the cutoff value of EMT for EP prediction. RESULTS: The incidence rate of EP was 2.95 % (92/3117). After adjustment for other factors in the logistic regression model, the incidence of EP decreased by 55 % with an EMT > 10 mm compared with an EMT ≤ 10 mm (odds ratio 0.450, 95 % confidence interval 0.296-0.684, P < 0.001). The EMT in the EP group was significantly thinner than that in the live birth (n = 2540) and spontaneous abortion (n = 485) groups (p < 0.017). The cutoff value of EMT for EP prediction was 10.65 mm, with a sensitivity of 59 % and a specificity of 63 %. CONCLUSION: A decreased risk of EP was found among the patients with an EMT > 10 mm prior to embryo transfer. A certain EMT is needed to reduce the incidence of EP.

Green tea extract as a cryoprotectant additive to preserve the motility and DNA integrity of human spermatozoa.

Cryopreservation impairs sperm quality and functions, including motility and DNA integrity. Antioxidant additives in sperm freezing media have previously brought improvements in postthawed sperm quality. Green tea extract (GTE) is widely considered as an excellent antioxidant, and its beneficial role has been proven in other human cells. This study aims to evaluate the GTE as a potential additive in cryopreservation media of human spermatozoa. In part one, the semen of 20 normozoospermic men was used to optimize the concentration of GTE that maintains sperm motility and DNA integrity against oxidative stress, induced by hydrogen peroxide (H2O2). Spermatozoa were treated with GTE at different concentrations before incubation with H2O2. In part two, the semen of 45 patients was cryopreserved with or without 1.0 ng ml-1 GTE. After 2 weeks, the semen was thawed, and the effect on sperm motility and DNA fragmentation was observed. Our data showed that GTE significantly protected sperm motility and DNA integrity against oxidative stress induced by H2O2when added at a final concentration of 1.0 ng ml-1. We found that the addition of 1.0 ng ml-1 GTE to cryopreservation media significantly increased sperm motility and DNA integrity (both P < 0.05). More interestingly, patients with high sperm DNA damage benefited similarly from the GTE supplementation. However, there was no significant change in the reactive oxygen species (ROS) level. In conclusion, supplementing sperm freezing media with GTE has a significant protective effect on human sperm motility and DNA integrity, which may be of clinical interest.

The effects of daily meteorological perturbation on pregnancy outcome: follow-up of a cohort of young women undergoing IVF treatment.

BACKGROUND: Human reproduction follows a seasonal pattern with respect to spontaneous conception, a phenomenon wherein the effect of meteorological fluctuations might not be unique. However, the effect of seasonal variations on patients who underwent in vitro fertilization (IVF) treatment is unclear. We aimed to evaluate the effects of meteorological variation on the pregnancy rate in a cohort undergoing IVF treatment by performing multivariable analyses. METHODS: We conducted a cohort study in a sub-tropical region with prominent seasonal variations (2005-2016). Women aged < 35 years who were treated with a long ovarian stimulation protocol and underwent fresh embryo transfer (ER) were included. Data on gonadotropin administration (CYCL), oocyte retrieval (OR), ER, and pregnancy outcomes were prospectively recorded. For each patient, the daily average of meteorological data (temperature, humidity, sunlight duration, solar radiation) was recorded from the date of CYCL to ER. Multiple logistic regression analysis adjusted for age, fertilization method, year of the cycle, gonadotropin dose, and transferred embryo grade was performed to determine the relationship between the meteorological parameters and clinical pregnancy. Patients with one successful cycle and one failed cycle were subtracted for a case-control subgroup analysis through mixed effect logistics regressions. Time-series analysis of data in the epidemic level was conducted using the distributed lag linear and non-linear models (DLNMs). RESULTS: There were 1029 fresh cycles in 860 women (mean age 31.9 ± 2.0 years). Higher mean temperature from CYCL to OR (adjusted odds ratio [aOR] 1.04; 95% confidence interval [CI] 1.01-1.07, P = 0.01) increased the odds of pregnancy, while OR to ER did not show any statistical significance. Compared to that in winter, the odds of becoming pregnant were higher during higher temperature seasons, summer and autumn (aOR 1.47, 95%CI 0.97-2.23, P = 0.07 (marginally significant) and aOR 1.73, 95%CI 1.12-2.68, P = 0.02, respectively). Humidity, sunlight duration, and solar radiation had no effect on the outcome. The subgroup analysis confirmed this finding. The time-series analysis revealed a positive association between temperature and relative risk for pregnancy. CONCLUSIONS: In IVF treatment, the ambient temperature variation alters the pregnancy rates; this aspect must be considered when obtaining patient consent for assisted conception.

A double-blinded, randomized placebo-controlled trial on the effect of traditional Chinese medicine formula Wuzi Yanzong pill on improving semen qualities in men with suboptimal parameters.

BACKGROUND: In Hong Kong, one of six couples is affected by subfertility problems. Male infertility contributes to half of the infertility cases. In male infertility, there is no effective treatment for patients with idiopathic infertility/poor semen parameters. Recent meta-analysis results suggest that a traditional Chinese medicine (TCM) formula - Wuzi Yanzong pill - showed a curative effect on male fertility. However, the heterogeneity of the studies could not draw a definitive conclusion on the therapeutic effect of this formula. The aim of this study is to conduct a well-designed randomized controlled trial to investigate the effect of TCM formula Wuzi Yanzong pill on improving semen qualities in men with suboptimal parameters. METHODS: This study is a double-blinded, randomized placebo-controlled trial conducted in a public hospital in Hong Kong. Participants will be randomized, using computer-generated random numbers, with a 1:1 ratio to either the Wuzi Yanzong pill formula group or the placebo group. Both groups will be administered the drugs for 12 weeks. Participants will have a total of four visits for their semen and blood assessments for a 6-month period, and we will follow up for another 6 months to record their conception outcome. The primary outcome is to compare the total motile sperm count, natural conception rate, and pregnancy outcome to those under placebo treatment. Secondary objectives are sperm functions and assisted reproductive technology outcome. DISCUSSION: To date, there are no studies using the disclosed Wuzi Yanzong formula or double-blinded, randomized trials. The Wuzi Yanzong TCM formula may provide a good clinical solution for subfertile males for which contemporary western medicine has no cure. Therefore, a well-designed randomized trial for evaluating the effect of Wuzi Yanzong TCM formula is urgently needed. TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR), ChiCTR-INR-17010790 . Registered on 27 February 2017. Centre for Clinical Research and Biostatistics - Clinical Trials Registry, CUHK_CCRB00548 . Registered on 27 February 2017.

Declined Natural Killer Cells Emerging in Women with Unexplained Recurrent Spontaneous Abortion and Further Reducing after Medical Therapy.

BACKGROUND: As focus grows on reproduction, the issue of Recurrent Spontaneous Abortion (RSA), especially for unexplained reasons (URSA), is grabbing more and more attention in gynecological immunology. We investigated the changes of peripheral lymphocyte subsets focusing on whether they had some relationship with development of URSA. METHODS: The percentage and absolute count of lymphocyte subsets (T cells, Th cells, Ts cells, B cells, NK cells) were simultaneously evaluated by flow cytometry in URSA patients (n = 48) and healthy controls (HC, n = 22). RESULTS: Significantly lower percentage and absolute counts of NKT cells and NK cells were observed in URSA compared to the HC. After medical therapy, an obviously increase was shown in the percentage of both T cells and B cells, whereas it presented a reduction in the percentage of NK cells. CONCLUSIONS: The flow cytometry test in T, B, NK cells is a method available to identify URSA patients from healthy women and to provide reference guides for clinical therapy.

Thrombophilia Markers in Patients with Recurrent Early Miscarriage.

BACKGROUND: This study was to investigate the association between thrombophilia and REM by the change of thrombophilia markers and to evaluate their contribution in diagnosis and treatment of REM. METHODS: 199 women with REM history were divided into two groups within the study group: 151 pregnant (REM-P) and 48 nonpregnant (REM-NP). In addition, 121 healthy age-matched women without REM history were divided into two groups of the control group: 75 pregnant (Control-P) and 46 nonpregnant (Control-NP). Lupus anticoagulant (LA), anticardiolipin antibodies (ACA), and anti-ß2-glycoprotein-I antibodies (anti-ß2GPI-ab) and coagulation-related factors such as protein S (PS), protein C (PC), anti-thrombin III (AT-III), and D-dimer were analysed. The prevalence of antiphospholipid antibodies (aPL) and the coagulation-related factors between groups were compared. RESULTS: The overall prevalence of aPL positivity in REM-P (14.57%) showed a difference compared with REM-NP (2.66%) but not for aCL, anti-ß2GPI-ab or LA alone. There were significant differences in the mean levels of protein S, protein C, and AT-III in REM-P. The mean values of protein C (90.3 ± 28.42%) and protein S (71.80 ± 24.68%) in the aPL positive study group were similar with that of the aPL negative study group (p = 0.406, p = 0.880). Comparing with the aPL negative study group, the mean value of AT-III (87.71 ± 21.84%) was significantly lower (p = 0.018), while the mean value of D-dimer (0.98 ± 1.1 mg/L FEU) was significantly higher (p = 0.013). CONCLUSIONS: We briefly address the role of the prevalence of aPL and the related coagulation factors for predicting a prethrombotic state in patients with REM. The results of the use of anticoagulants for treating REM are encouraging.