A study of application effects of next-generation sequencing based preimplantation genetic testing for aneuploidies in advanced maternal age women.

OBJECTIVE: To investigate if next-generation sequencing-based preimplantation genetic testing for aneuploidies could improve pregnancy outcomes in women of advanced maternal age. MATERIALS AND METHODS: A retrospective analysis. The clinical data of 1099 couples treated in the First Medical Center of the Chinese PLA General Hospital from January 2019 to December 2021 were analyzed. They were divided into two groups based on whether they underwent a Next-generation sequencing-based preimplantation genetic test for aneuploidies. We analyzed and compared the biochemical pregnancy rate, clinical pregnancy rate, abortion rate, and live birth rate between the two groups. RESULTS: The Preimplantation genetic testing for aneuploidies (PGT-A) group was associated with higher rate of biochemical pregnancy and clinical pregnancy than the non-PGT-A group, which were 63.9% vs. 56.4% (P = 0.009) and 54.4% vs. 45.6% (P < 0.001), respectively. The abortion rate was significantly lower in the PGT-A group compared to the non-PGT-A group (2.3% vs. 14.7%, P < 0.001). In addition, the live birth rate was significantly higher in the PGT-A group compared to the non-PGT-A group (52.1% and 30.9%, respectively, P < 0.001). CONCLUSION: Next-generation sequencing-based preimplantation genetic testing for aneuploidies significantly improved the pregnancy outcomes in women of advanced maternal age.

Clinical Application of Double Ovulation Stimulation in Patients with Diminished Ovarian Reserve and Asynchronous Follicular Development Undergoing Assisted Reproduction Technology.

OBJECTIVE: This study aimed to compare the clinical effects of double ovulation stimulation (DouStim) applied during the follicular and luteal phases with the antagonist protocol in patients with diminished ovarian reserve (DOR) and asynchronous follicular development undergoing assisted reproductive technology (ART). METHODS: The clinical data of patients with DOR and asynchronous follicular development receiving ART from January 2020 to December 2021 were retrospectively analyzed. The patients were divided into two groups according to their ovulation stimulation protocol: DouStim group (n=30) and antagonist group (n=62). Assisted reproduction and clinical pregnancy outcomes were compared between the two groups. RESULTS: In the DouStim group, the number of oocytes retrieved, metaphase II (MII) oocytes, two-pronuclei (2PN), day 3 (D3) embryos, D3 high-quality embryos as well as blastocyst formation, implantation, and human chorionic gonadotropin-positive rates were significantly greater than those in the antagonist group (all P<0.05). No significant differences were found in MII, fertilization, or continued pregnancy rates at the first frozen embryo transfer (FET), in-vitro fertilization (IVF) cancellation, or early medical abortion rates between the groups (all P>0.05). Except for the early medical abortion rate, the DouStim group generally had favorable outcomes. In the DouStim group, the dosage and duration of gonadotropin and the fertilization rate were significantly greater in the first ovulation stimulation induction than in the second ovulation stimulation induction (P<0.05). CONCLUSION: The DouStim protocol efficiently and economically obtained more mature oocytes and high-quality embryos for patients with DOR and asynchronous follicular development.

Clinical Characteristics and Influence Factor Analysis of ABO Typing Discrepancy Among Patients in a Tertiary Hospital.

BACKGROUND: An accurate interpretation of the ABO blood group of an individual is of utmost importance to ensure patient safety and good transfusion practices. The aim of the study was to determine the incidence and causes of ABO typing discrepancies among patients and analysis of the clinical characteristics as well. METHODS: A retrospective observational study was carried out in the Department of Laboratory Medicine in the General Hospital of Chinese PLA from March 2018 to December 2020. Records of patients were collected and analyzed for frequency, clinical characteristics, and influencing factors of ABO typing discrepancies. RESULTS: There were 132 ABO typing discrepancies patients (57 females, 75 males), aged from 3 to 84 (50.3 ± 19.3) years, with history of operation 89 cases (67.42%) and blood transfusion 23 cases (17.42%). In the control group, there were 142 cases (63 females and 79 males), aged 10 - 78 years (50.5 ± 15.55), with operation history 68 cases (47.88%) and blood transfusion history 2 cases (1.40%). Among the inconsistent blood types group, there were 59 cases (44.7%) type B, 38 cases (28.8%) type A, 28 cases (21.2%) type AB, and 7 cases (5.3%) type O. There were 21 cases (15.9%) in the department of hematology, 15 cases (11.36%) in the department of orthopedics, 14 cases (10.6%) in the department of hepatobiliary surgery, 11 cases (8.33%) in the department of general surgery, 11 cases (8.33%) in the department of vascular surgery, and less than 5% in other departments. The common cause of ABO typing discrepancies was due to low/weak affinity antibody (3.79%), low/weak affinity antibody B (18.18%), weak A antigen (3.03%), weak B antigen (12.12%), hematopoietic stem cell transplantation (10.60%), irregular antibody (46.70%), subtype (3.79%), and cold agglutination (1.51%). The diseases in the ABO discrepancy group mainly included hematological disorders, malignant tumors, osteoarthritis and so on. Binary logistic regression showed that hematological disorders, malignant tumors, history of operation (p = 0.01), and history of blood transfusion (p = 0.017) were the influencing factors of ABO typing discrepancies. CONCLUSIONS: It is possible that the antibody of ABO blood group system could not be detected. The independent influence factors were hematological disorders, malignant tumor, blood transfusion history and operation history for ABO typing discrepancies. It was necessary to analyze the causes correctly and judge the correct blood group by serological methods.

The Differential and Dynamic Progression of Hepatic Inflammation and Immune Responses During Liver Fibrosis Induced by Schistosoma japonicum or Carbon Tetrachloride in Mice.

Liver fibrosis can result from various causes and could progress to cirrhosis and cancer; however, there are no effective treatments due to that its molecular mechanism is unclear. liver fibrosis model made by Schistosoma japonicum (S. japonicum) infection or Carbon tetrachloride (CCl4) intraperitoneal injection is a conventional model used in liver fibrosis-related studies for mechanism or pharmaceutical research purposes. But the differences in the pathological progression, immune responses and the underlying mechanism between the two liver fibrosis model have not been carefully compared and characterized, which hinders us from correctly understanding and making better use of the two models. In the present study, the pathological changes to the liver, and the cytokines, inflammatory factors, macrophages, and lymphocytes subsets involved were analyzed in the liver fibrosis model of S. japonicum infection or CCl4 intraperitoneal injection. Additionally, the pathological progression, immune responses and the underlying injury mechanism in these two models were compared and characterized. The results showed that the changing trend of interleukin-13 (IL-13), transforming growth factor beta (TGF-ß), inflammatory factors, and M1, M2 macrophages, were consistent with the development trend of fibrosis regardless of whether liver fibrosis was caused by S. japonicum or CCl4. For lymphocyte subsets, the proportions of CD3+ T cells and CD4+ T cells decreased gradually, while proportion of CD8+ T cells peaked at 6 weeks in mice infected with S. japonicum and at 12 weeks in mice injected with CCl4. With prolonged S. japonicum infection time, Th1 (CD4+IFN-γ+) immunity converted to Th2 (CD4+IL-4+)/Th17 (CD4+IL-17+) with weaker regulatory T cell (Treg) (CD4+CD25+FOXP3+) immunity. However, in liver fibrosis caused by CCl4, Th1 cells occupied the dominant position, while proportions of Th2, Th17, and Treg cells decreased gradually. In conclusion, liver fibrosis was a complex pathological process that was regulated by a series of cytokines and immune cells. The pathological progressions and immune responses to S. japonicum or CCl4 induced liver fibrosis were different, possibly because of their different injury mechanisms. The appropriate animal model should be selected according to the needs of different experiments and the pathogenic factors of liver fibrosis in the study.

Whole­exome sequencing analysis of products of conception identifies novel mutations associated with missed abortion.

Missed abortion (MA) refers to a pregnancy in which there is fetal demise without outside intervention, and additionally no uterine activity that may expel the product of conception (POC) prior to 20 weeks of gestation. Chromosomal abnormalities are the primary cause of MA and single gene defects in the POC may additionally be associated with MA; however, few studies have been conducted on the identification of mutations by whole­exome sequencing. In the present study, 19 unrelated MA POCs were collected and whole­exome sequencing was performed on the POC. Bioinformatics analysis was performed on sequence variants from a list of 286 selected candidate genes that were associated with early embryonic lethality and MA. A total of 36 sequence variants in 32 genes potentially associated with MA were identified in 15 out of 19 patients. Gene Ontology analysis suggested that these genes were enriched in biological processes in early embryonic development, including 'chordate embryonic development', 'cell proliferation' and 'forebrain development'. Further strict in silico bioinformatics analysis predicted that the LIM domain­binding protein 1 (c.662C>T; p.S221L) variant was a highly pathogenic variant. In conclusion, the results of the present study provide researchers and clinicians with a better understanding of the etiology and molecular mechanism of human embryonic lethality and MA.

Intention of having a second child among infertile and fertile women attending outpatient gynecology clinics in three major cities in China: a cross-sectional study.

STUDY QUESTION: What is the intention to have a second child among women attending outpatient gynecology clinics in three major cities in China? SUMMARY ANSWER: In total, 69.3% of the participants expressed the intention to have a second child and this was related to infertility status, pronatalist attitudes, and sociodemographic factors. WHAT IS KNOWN ALREADY: In 2016, the new universal two-child policy was introduced in China enabling all Chinese couples to have a second child. A government-led national survey revealed that the majority of women included under the policy would be 35 years old and older and thus would be at higher risk of infertility. Previous studies found that fertility intention differs by infertility status. STUDY DESIGN SIZE DURATION: A cross-sectional survey was performed to examine the intention of having a second child and its associated factors among infertile and fertile women attending gynecology outpatient clinics in three major cities in China. Clinical nurses approached eligible women in person while waiting for their consultations. Recruitment and data collection were conducted from April to August 2016. PARTICIPANTS/MATERIALS SETTING METHODS: The survey involved four gynecology outpatient clinics in Beijing, Shenzhen, and Hohhot. Married women aged 20-45 years who were seeking outpatient gynecology care for non-malignant problems were invited to participate. MAIN RESULTS AND THE ROLE OF CHANCE: Data from 974 women were included in the analysis. A total of 69.3% of the women expressed the intention to have a second child, and infertile women were more likely to want a second child compared to fertile women (76.6% vs 61.9%, respectively; P < 0.001). Greater ideal parity facilitated the intention for a second child in both groups, while pronatalist attitudes (meaning that they preferred to have their first childbirth at a younger age and attached greater significance to traditional childbearing beliefs), unexplained infertility, presence of a living child and religious affiliation were associated with greater intention among infertile women. In contrast, in the fertile group, older age, full-time work and lower confidence in achieving parity goals diminished the intention for a second child. Although infertile women displayed greater agreement with pronatalist attitudes and desired a higher ideal parity, they had less confidence in achieving their parity goals than their fertile counterparts. LIMITATIONS REASONS FOR CAUTION: In addition to self-report and self-selection bias, our participants were recruited from urbanized areas and were more educated than the general population. Owing to the extremely busy environment in the clinics, difficulties were encountered in keeping track of the number of women whom the nurses approached, and the response rate was therefore unavailable. WIDER IMPLICATIONS OF THE FINDINGS: With the introduction of the universal two-child policy, there is a need to enhance fertility awareness and to encourage reproductive life planning, as well as to lower the cost of childcare, in order to increase the birth rate in China. Effort is required to make childbearing more compatible with current employment, career and educational development, the burdens of family care (e.g. for elderly parents), social environments and cultural expectations. This is particularly relevant for families who already have a child, as our findings show that their hesitation toward a second child was largely related to difficulties with extra childcare within the woman's current work and family life. STUDY FUNDING/COMPETING INTERESTS: This study did not receive any funding. The authors declared no competing interests. TRIAL REGISTRATION NUMBER: Not applicable.

The therapeutic potential of umbilical cord mesenchymal stem cells in mice premature ovarian failure.

Mesenchymal stem cells, which are poorly immunogenic and have potent immunosuppressive activities, have emerged as promising cellular therapeutics for the treatment of several diseases. Mesenchymal-like cells derived from Wharton's Jelly, called umbilical cord matrix stem cells (UCMSCs), reportedly secrete a variety of cytokines and growth factors, acting as trophic suppliers. Here, we used UCMSCs to treat premature ovarian failure (POF). Ovarian function was evaluated by ovulation and the number of follicles. Apoptosis of the granulosa cells (GC) was analyzed by TUNEL staining. We found that after transplantation of the UCMSCs, apoptosis of cumulus cells in the ovarian damage model was reduced and the function of the ovary had been recovered. The sex hormone level was significantly elevated in mice treated with UCMSCs. The number of follicles in the treated group was higher than in the control group. Our results demonstrate that UCMSCs can effectively restore ovary functionality and reduce apoptosis of granulosa cells. We compared the RNA expression of the UCMSCs treated group with the POF model and wild-type control group and found that the UCMSC group is most similar to the wild-type group. Our experiments provide new information regarding the treatment of ovarian function failure.