Embryo development is impaired by sperm mitochondrial-derived ROS

Background Basal energetic metabolism in sperm, particularly oxidative phosphorylation, is known to condition not only their oocyte fertilising ability, but also the subsequent embryo development. While the molecular pathways underlying these events still need to be elucidated, reactive oxygen species (ROS) could have a relevant role. We, therefore, aimed to describe the mechanisms through which mitochondrial activity can influence the first stages of embryo development. Results We first show that embryo development is tightly influenced by both intracellular ROS and mitochondrial activity. In addition, we depict that the inhibition of mitochondrial activity dramatically decreases intracellular ROS levels. Finally, we also demonstrate that the inhibition of mitochondrial respiration positively influences sperm DNA integrity, most likely because of the depletion of intracellular ROS formation. Conclusion Collectively, the data presented in this work reveals that impairment of early embryo development may result from the accumulation of sperm DNA damage caused by mitochondrial-derived ROS.

Staged Treatment with Chinese Herbal Medicine in a Controlled Ovarian Hyperstimulation of In Vitro Fertilization-Embryo Transfer Programme / 中医杂志

It is believed that kidney deficiency is the fundamental pathogenesis while liver constraint and spleen deficiency， and disharmony of chong （thorough vessel， 冲脉）and ren （conception vessel， 任脉） is the key pathogenesis of infertility patients who adopted controlled ovarian hyperstimulation of in vitro fertilization-embryo transfer （IVF-ET） programme. Therefore， the method of tonifying the kidneys dominantly and treating the liver and spleen simultaneously is proposed， and Chinese herbal medicine is suggested to be used in adjuvant treatment of staged IVF-ET controlled ovrian hyperstimulation. In the regulation stage， modified Liuwei Dihuang Pill （六味地黄丸） can be used to tonify kidney and supplement essence， fortify spleen and nourish liver； in the ovulation promotion stage， modified Wenjing Decoction （温经汤） should be used to warm kidney and assist yang， dispel stasis and nourish blood； in the pre-transplantation endothelial preparation stage， modified Shenling Baizhu Powder （参苓白术散） is suggested to fortify spleen and replenish qi， invigorate blood and resolve stasis； after the transplantation stage， modified Shoutai Pill （寿胎丸） or Taiyuan Decoction （胎元饮） can be taken to fortify spleen and tonify kidney， benefit qi and nourish blood.

Morpho-anatomy of in vitro germination and cryopreservation of the orchid Cattleya crispa (Orchidaceae Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae) / Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae)

Introduction: Cattleya crispa is an ornamental epiphytic orchid with geographic distribution restricted to the Brazilian Atlantic Forest. Due to predatory extractivism and human-induced habitat loss, this species appears on the Red List of Brazilian Flora. Objective: To characterize morpho-anatomical aspects regarding germination and post-seminal development from C. crispa seeds; as well as studying the effect of cryopreservation on these seeds. Methods: We used light microscopy and electron microscopy to describe the microstructure of a 100 ripe seeds. We evaluated seed viability, seed germination, survival rate and protocorm weight in cryopreserved and non-cryopreserved material, with four replicas per treatment using 20 mg of plant material. Results: The seeds are fusiform, whitish yellow with a length from 700 to 900 µm and a water content of 5 %. Germination began seven days after sowing, the formation of the globular protocorm at 30 days and the formation of the seedling occurred 150 days. The persistent seed coat can compress the protocorm and cause it to collapse. The cryopreserved seeds presented 87.15 % viability, 78.32 % germination, 8.48 % survival and protocorms with 104.27 mg five months after sowing. Data wasn't different to non-cryopreserved seeds. Conclusions: The cryocapability of the seeds shows that cryopreservation can be used for long-term conservation. The results of this work contribute to the overall biology of C. crispa and to the propagation and storage of genetic material for conservation purposes.

Introducción: Cattleya crispa es una orquídea epífita ornamental con distribución geográfica restringida a la Mata Atlántica brasileña. Debido al extractivismo depredador y a la pérdida de hábitat inducida por el hombre, esta especie aparece en la Lista Roja de la Flora Brasileña. Objetivo: Caracterizar aspectos morfoanatómicos de la germinación y desarrollo inicial de semillas de C. crispa; así como estudiar el efecto de la criopreservación de estas semillas. Métodos: Utilizamos microscopía óptica, microscopía electrónica de barrido y microscopía electrónica de transmisión para describir la microestructura en 100 semillas maduras. Evaluamos la viabilidad de la semilla, la germinación de la semilla, la tasa de supervivencia y el peso de los protocormos en el material criopreservado y no criopreservado, con cuatro réplicas por tratamiento de 20 mg de material vegetal. Resultados: Las semillas son fusiformes, amarillo blanquecinas, con una longitud de 700 a 900 µm y un contenido de agua del 5 %. La germinación comenzó siete días después de la siembra, la formación del protocormo globular a los 30 días y la formación de la plántula a los 150 días. La cubierta de semilla persistente puede comprimir el protocormo y provocar su colapso. Las semillas criopreservadas presentaron 87.15 % de viabilidad, 78.32 % de germinación, 8.48 % de supervivencia y protocormos con 104.27 mg a los cinco meses de la siembra. Los datos no fueron diferentes a las semillas no criopreservadas. Conclusiones: La capacidad criogénica de las semillas muestra que la crioconservación puede utilizarse para la conservación a largo plazo. Los resultados de este trabajo contribuyen a la biología general de C. crispa y a la propagación y almacenamiento de material genético con fines de conservación.

Development of the Goose Tongue Filiform Papillae: Could it be Tooth-Like Sense Organs? / Desarrollo de las Papilas Filiformes de la Lengua de Ganso: ¿Podrían ser Órganos Sensoriales Parecidos a los Dientes?

SUMMARY: The geese's tongue filiform papillae are particularly long, and exhibit the same morphology of a tooth, evoking the lingual teeth of several fishes. In adult animals, they contain numerous mechanical Herbst's corpuscles but no taste buds. In the embryo, they appear since stage 38 and acquire their definitive shape between stages 38 and 42. They express several proteins associated with mammalian tooth development (BMP4, β-catenin, SHH, PITX2, PAX9), also known to be linked to parrot's pseudoteeth and goose's denticulations development. Neurofilaments are early present in the papillae primordia, and appear particularly numerous in adult papillae. Our results suggest that these papillae constitute a mechanical organ with a « tooth shape » derived from ancestral odontodes, whose development is controlled by numerous genes involved in classical odontogenesis.

Las papilas filiformes de la lengua de los gansos son particularmente largas y exhiben la morfología de un diente, evocando los dientes linguales presentes en varios peces. En los animales adultos, contienen numerosos corpúsculos de Herbst mecánicos, aunque una ausencia de papilas gustativas. En el embrión, aparecen a partir del estadio 38 y adquieren su forma definitiva entre los estadios 38 y 42. Expresan varias proteínas asociadas al desarrollo dentario de los mamíferos (BMP4, β-catenina, SHH, PITX2, PAX9), también conocidas por estar asociadas al desarrollo de pseudodientes en el loro y denticulaciones en el ganso. Los neurofilamentos están presentes tempranamente en los primordios de las papilas y aparecen particularmente numerosos en las papilas adultas. Nuestros resultados sugieren que estas papilas constituyen un órgano mecánico con «forma de diente» derivado de odontoides ancestrales, cuyo desarrollo está controlado por numerosos genes implicados en la odontogénesis clásica.

Consideraciones éticas y científicas del inicio de la vida sobre el aborto eugenésico en enfermedades genéticas incapacitantes en Perú

El artículo expone conceptos actuales biológicos, así como algunos planteamientos filosóficos acerca del inicio de la vida, que se examinarán en relación con la necesidad del aborto eugenésico. Se presenta el concepto de "exdurantismo", en el que la combinación del nuevo genoma nuclear y mitocondrial hacen único al individuo; además, se concibe al complejo genómico como "integrador somático" que dirige el desarrollo embrionario, y se muestra la problemática en el caso de las violaciones que provocan embarazos, el aborto eugenésico en enfermedades genéticas incapacitantes o la utilización de embriones posterior a la fertilización in-vitro. Se plantea que la dignidad de la persona comienza desde la concepción. Frente a la detección de una enfermedad genética, grave o letal, se debe ofrecer soporte económico y social, de diagnóstico y tratamiento; además, desde la salud pública, una mayor inversión para plantear estrategias de tamizaje, diagnóstico, manejo e investigación.

The article presents current biological concepts, as well as some philosophical approaches to the beginning of life, which will be examined in relation to the need for eugenic abortion. The concept of "exdurantism" is presented, in which the combination of the new nuclear and mitochondrial genome makes the individual unique; furthermore, the genomic complex is conceived as a "somatic integrator" that directs embryonic development, and the problematic is shown in the case of rape that causes pregnancies, eugenic abortion in incapacitating genetic diseases or the use of embryos after in-vitro fertilization. The dignity of the person begins at conception. When a serious or lethal genetic disease is detected, economic and social support, diagnosis and treatment should be offered; in addition, public health should invest more in screening, diagnosis, management and research strategies.

O artigo apresenta conceitos biológicos atuais, bem como algumas abordagens filosóficas sobre o início da vida, que serão examinados em relação à necessidade do aborto eugênico. É apresentado o conceito de "exdurantismo", no qual a combinação do novo genoma nuclear e mitocondrial torna o indivíduo único; além disso, o complexo genômico é concebido como um "integrador somático" que dirige o desenvolvimento embrionário, e é mostrada a problemática no caso de estupro que resulta em gravidez, aborto eugênico em doenças geneticamente incapacitantes ou o uso de embriões após a fertilização in vitro. Argumenta-se que a dignidade da pessoa começa na concepção. Diante da detecção de uma doença genética, grave ou letal, devem ser oferecidos apoio econômico e social, diagnóstico e tratamento, bem como maior investimento em saúde pública em estratégias de triagem, diagnóstico, gestão e pesquisa.

Post-modern bioethical stresses destination of surplus embryos: Brazilian analysis and brief comparisons

The objective of the present study is to point out tensions of the theoretical/practical universe that Bioethics is facing in Brazil, in the search for a praxis for the destination of surplus embryos. We consider that Bioethics analyzes the implications of such practices in society and in relationships between individuals. Brief data from other countries were presented to compare the Brazilian situation progressively from the conceptual point of view and the adoption of measures. The research is a scoping review on the main points that have been hindering the progress of discussions on the subject and consequently the respective solution. The legal status of the embryo was described from several perspectives and theories, with the resulting proposals for the destination of surplus embryos in their positive and negative aspects. The tensions of Bioethics were presented in the context of post-modernity and the consequent social and moral plurality, together with the difficulties of identifying a secular bioethical morality. In the end, we conclude the possibility of proclaiming a consensus on the destination of surplus embryos based on secular morality, supported by the figure of the "moral strangers".

El objetivo del presente estudio es señalar las tensiones del universo teórico/práctico que la bioética enfrenta en Brasil, en la búsqueda de una praxis para el destino de los embriones sobrantes. Consideramos que la bioética analiza las implicaciones de tales prácticas en la sociedad y en las relaciones entre los individuos. Se presentaron breves datos de otros países para comparar progresivamente la situación brasileña desde el punto de vista conceptual y de la adopción de medidas. La investigación es una revisión del alcance de los principales puntos que han obstaculizado el avance de las discusiones sobre el tema y, en consecuencia, la respectiva solución. Se describió el estatuto jurídico del embrión desde diversas perspectivas y teorías, con las consiguientes propuestas para el destino de los embriones sobrantes en sus aspectos positivos y negativos. Se presentaron las tensiones de la bioética en el contexto de la posmodernidad y la consecuente pluralidad social y moral, junto con las dificultades de identificar una moral bioética laica. Al final, se concluye la posibilidad de proclamar un consenso sobre el destino de los embriones sobrantes basado en una moral laica, apoyada en la figura de los "extraños morales".

O objetivo do presente estudo é destacar as tensões do universo teórico/prático que a Bioética está enfrentando no Brasil, na busca de uma praxis para a destinação de embriões excedentes. Nós consideramos que a Bioética analisa as implicações de tais práticas na sociedade e nas relações entre indivíduos. Dados resumidos de outros países são apresentados para comparar a situação brasileira progressivamente de um ponto de vista conceitual para a adoção de medidas. A pesquisa é uma revisão de escopo sobre os pontos principais que vem atrapalhando o andamento das discussões sobre o assunto e consequentemente a solução respectiva. O status legal do embrião foi descrito a partir de diversas perspectivas e teorias, com as propostas resultantes para a destinação dos embriões excedentes em seus aspectos positivos e negativos. As tensões da Bioética foram apresentadas no contexto da pós-modernidade e a consequente pluralidade social e moral, juntamente com as dificuldades de identificar uma moralidade bioética secular. Ao final, nós concluímos pela possibilidade de proclamar um consenso sobre a destinação de embriões excedentes baseado na moralidade secular, apoiado pela figura dos "estranhos morais".

Cell-free DNA in Embryo Culture Media as Non-invasive Biomarker of the Quality of Embryo Cleavage

Background: The success of in vitro-fertilization (IVF) cycles is determined in large part by the quality of embryo cleavage, which in turn, is dependent on the quality of the embryo culture media (CM). Many factors can influence the quality of embryo CM, one of which is the levels of Cell Free Deoxyribonucleic acid (DNA). Understanding the association between Cell-free DNA levels in embryo CM and the quality of embryo cleavage could help improve the quality of IVF techniques. Methods: This prospective study was conducted with 96 spent CM from patients undergoing IVF cycle, in order to determine relationships of Cell-free DNA levels in embryo CM with embryo cleavage quality on day 3. After intracytoplasmic sperm injection (ICSI), 48 embryos were evaluated on day 3 of their development, according to their cell number. Day 2 and day 3 CM corresponding to each one of the embryos was analyzed, by quantitative PCR, for estimation of Cell-free DNA levels. Results: The results revealed a significant increase in Cell-free DNA levels on day 2 CM corresponding to 4 to 6 cell embryos compared to those corresponding to 7 to 8 cell embryos (p=0.04). As for day 3 CM, the results showed no significant difference between the Cell-Free DNA levels in CM of 7-8 and those of 4-6 cell embryos (p=0.4). Also, cell free DNA levels in embryo CM, were significantly higher on day 2 compared to day 3 for both 7-8 and 4-6 cell embryos (p=0.03; p=0.04). Conclusion: We conclude that cell-free DNA levels in CM might be associated with delayed embryo cleavage.

Is the length of time between endometrial scratching and embryo transfer important for pregnancy success? An observational study

SUMMARY OBJECTIVE: This study sought to evaluate the influence of time (early <90 days and late >90 days) and endometrial injury on pregnancy success. METHODS: This is a retrospective study in which all infertile women who underwent at least one in vitro fertilization cycle at Clinica Gera between 2010 and 2015 were considered for inclusion. We included patients with a normal ovarian reserve and regular menses at intervals of up to 30 days. A total of 315 patient files were reviewed, and the study group was composed of patients who faced fertility issues and had male-caused infertility or idiopathic infertility. Also, women with male or unknown cause of infertility who have performed endometrial biopsy and have undergone embryo transfer up to 180 days after this procedure between 2010 and 2015 were included. The patients were divided into two groups according to the interval between biopsy and embryo transfer: group 1 (early—an interval of <90 days) and group 2 (late—an interval of >90 days and up to 180 days). RESULTS: The results were superior for the group with an interval of less than 90 days relative to the group with an interval of more than 90 days (p<0.04). The pregnancy rates for group 1 and group 2 were 58.5% and 43.4%, respectively. The odds ratio for pregnancy success was 1.63 (95% confidence interval: 1.04 to 2.55). CONCLUSION: The early transfer of embryos (<90 days) may produce better results with a high rate of pregnancy. Further studies are necessary to identify the mechanism involved in this phenomenon.

Autophagy Regulated by Chinese Medicine Monomers in Embryo Implantation： A Review / 中国实验方剂学杂志

Embryo implantation is one of the most critical steps in the reproductive process. The failure of embryo implantation to continue development is one of the important reasons leading to infertility. The success of embryo implantation depends on the high receptivity of endometrium and the embryo with implantation ability. Autophagy is a process in which cytoplasm， organelles， and inclusions are absorbed by double-membrane vesicles and transported to lysosomes for degradation and recycling， which is a way to maintain the homeostasis. A large amount of evidence have shown that autophagy plays an important role in all aspects of embryo implantation. Based on this， this paper explored the relationship between autophagy and endometrial receptivity and embryo implantation ability. According to the latest research progress， this paper combed 5 mechanisms （promotion of decidualization of endometrial stromal cells， promotion of apoptosis， regulation of hormone levels， coordination of inflammation， and promotion of ovulation） of 14 kinds of Chinese medicine monomers， including emodin， catalpol， paeoniflorin， resveratrol， folic acid， zearalenone， curcumin， wogonin， quercetin， chrysin， berberine， apigenin， phisetine， and kaempferol， in regulating different links of autophagy intervention in embryo implantation. This paper is expected to provide references and ideas for future Chinese medicine monomers to improve the success rate of embryo implantation.

Application progress of mobile health in nursing of patients with infertility / 中国实用护理杂志

This article firstly reviews the current application status of mobile health based on social networking media, mobile health applications, wearable devices in infertility patients′nursing. Then, this paper analyzes the application effects of mobile health in the five aspects of treatment process management, health education, medication management, lifestyle management and psychological care for infertility patients based on the literature. The existing problems are analyzed and prospected on this basis, in order to provide a reference for the application of mobile health to infertility patients′nursing in China.

Clinical Efficacy of Oral Dydrogesterone Alone in Natural Cycle Frozen-thawed Embryo Transfer / 中山大学学报(医学科学版)

ObjectiveTo investigate the clinical efficacy of oral dydrogesterone alone for luteal phase support in natural cycle frozen-thawed embryo transfer （NC-FET）. MethodsThe clinical data of 1 530 NC-FET cycles enrolled in our Reproductive Center from January 2019 to September 2021 were retrospectively analyzed. According to different luteal support protocols， the patients were divided into oral dydrogesterone alone （group A， n=524）， vaginal progesterone soft capsules （group B， n=401） and vaginal progesterone soft capsules combined with dydrogesterone （group C， n=605）. The clinical outcomes and cost-effectiveness ratio were compared among the three groups. The primary outcome was live birth rate. ResultsThe live birth rate was 43.13% （226/524） in group A， 39.15% （157/401） in group B， and 42.64% （258/605） in group C. There was no statistical difference among the three groups （P > 0.05）. No statistical difference was observed in the HCG positive rate， implantation rate， biochemical pregnancy rate， clinical pregnancy rate， spontaneous miscarriage rate， ectopic pregnancy rate， twin delivery rate， premature delivery rate and newborn weight among the three groups （P>0.05）. Logistic regression analysis revealed that the three luteal support regimens did not affect live birth rate. Pharmacoeconomic analysis showed that taking group B as a reference， the cost increased by 19 227.30 yuan for every 1% increase in live birth rate in group A. ConclusionsIn NC-FET cycle， oral dydrogesterone alone can achieve the same clinical outcomes as vaginal progesterone soft capsules and vaginal progesterone soft capsules combined with dydrogesterone. Compared with that of progesterone soft capsules， the cost of oral dydrogesterone alone is increased， a large sample and multicenter prospective study is needed to further confirm our results.

Comparison of the Effects of Single Sperm Cryopreservation and Conventional Cryopreservation on Embryo Culture and Pregnancy in Patients Undergoing Micro-TESE / 中山大学学报(医学科学版)

ObjectiveTo compare the effects of single sperm cryopreservation and conventional cryopreservation on embryo culture and pregnancy in patients undergoing microdissection testicular sperm extraction （micro-TESE）. MethodsA retrospective analysis was done on the patients who underwent micro-TESE at the Reproductive Medicine Center in the Sixth Affiliated Hospital of Sun Yat-Sen University between January 2018 and December 2021. The single sperm cryopreservation group included 39 patients undergoing single sperm cryopreservation and 307 MII oocytes. The conventional cryopreservation group included 39 patients undergoing conventional cryopreservation and 410 MII oocytes. Propensity score matching （PSM） was performed to balance the selection bias. The fertility rate， embryo culture and pregnancy of these two groups were compared. ResultsThere was no statistical difference in age， body mass index （BMI）， years of infertility， basal FSH， basal LH， basal E2， AMH， AFC， number of oocytes retrieved and number of transferred embryos between the two groups （P>0.05）. No significant difference was found in fertilization rate （65.8% vs. 60.49%）， available embryo rate （67.82% vs. 58.87%） and high-quality embryo rate （70.80% vs. 75.34%）. The single sperm cryopreservation group had significantly lower clinical pregnancy rate than conventional cryopreservation group （45.45% vs. 70.0%， P=0.049）. ConclusionIf the patients undergoing micro-TESE have very few sperms， single sperm cryopreservation could be an effective choice to increase the utilization of each sperm and ensure the subsequent sperm retrieval after thawing， but the clinical pregnancy rate is decreased. Due to the limited number of cases， we need a large additional number of cases to observe and analyze.

Human 8-cell embryos enable efficient induction of disease-preventive mutations without off-target effect by cytosine base editor

Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.

Effects of Zishen Yutai Pills on in vitro Fertilization-Embryo Transfer Outcomes in Patients with Diminished Ovarian Reserve: A Prospective, Open-Labeled, Randomized and Controlled Study / 中国结合医学杂志

OBJECTIVE@#To explore the effects of Zishen Yutai Pills (ZYPs) on the quality of oocytes and embryos, as well as pregnancy outcomes in patients with diminished ovarian reserve (DOR) receiving in vitro fertilization-embryo transfer (IVF-ET). The possible mechanisms, involving the regulation of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), were also investigated.@*METHODS@#A total of 120 patients with DOR who underwent their IVF-ET cycle were randomly allocated to 2 groups in a 1:1 ratio. The patients in the treatment group (60 cases) received ZYPs from the mid-luteal phase of the former menstrual cycle by using gonadotropin-releasing hormone (GnRH) antagonist protocol. The patients in the control group (60 cases) received the same protocol but without ZYPs. The primary outcomes were the number of oocytes retrieved and high-quality embryos. Secondary outcomes included other oocyte or embryo indices as well as pregnancy outcomes. Adverse events were assessed by comparison of the incidence of ectopic pregnancy, pregnancy complications, pregnancy loss, and preterm birth. Contents of BMP15 and GDF9 in the follicle fluids (FF) were also quantified with enzyme-linked immunosorbent assay.@*RESULTS@#Compared with the control group, the numbers of oocytes retrieved and high-quality embryos were significantly increased in the ZYPs group (both P<0.05). After treatment with ZYPs, a significant regulation of serum sex hormones was observed, including progesterone and estradiol. Both hormones were up-regulated compared with the control group (P=0.014 and 0.008), respectively. No significant differences were observed with regard to pregnancy outcomes including implantation rates, biochemical pregnancy rates, clinical pregnancy rates, live birth rates, and pregnancy loss rates (all P>0.05). The administration of ZYPs did not increase the incidence of adverse events. The expressions of BMP15 and GDF9 in the ZYPs group were significantly up-regulated compared with the control group (both P<0.05).@*CONCLUSIONS@#ZYPs exhibited beneficial effects in DOR patients undergoing IVF-ET, resulting in increments of oocytes and embryos, and up-regulation of BMP15 and GDF9 expressions in the FF. However, the effects of ZYPs on pregnancy outcomes should be assessed in clinical trials with larger sample sizes (Trial reqistration No. ChiCTR2100048441).

Effect of advanced paternal age on reproductive outcomes in IVF cycles of non-male-factor infertility: a retrospective cohort study / 亚洲男科学杂志(英文版)

Advanced paternal age has been overlooked, and its effect on fertility remains controversial. Previous studies have focused mainly on intracytoplasmic sperm injection (ICSI) cycles in men with oligozoospermia. However, few studies have reported on men with semen parameters within reference ranges. Therefore, we conducted a retrospective cohort study analyzing the reproductive outcomes of couples with non-male-factor infertility undergoing in vitro fertilization (IVF) cycles. In total, 381 cycles included were subgrouped according to paternal age (<35-year-old, 35-39-year-old, or ≥40-year-old), and maternal age was limited to under 35 years. Data on embryo quality and clinical outcomes were analyzed. The results showed that fertilization and high-quality embryo rates were not significantly different (all P > 0.05). The pregnancy rate was not significantly different in the 35-39-year-old group (42.0%; P > 0.05), but was significantly lower in the ≥40-year-old group (26.1%; P < 0.05) than that in the <35-year-old group (40.3%). Similarly, the implantation rate significantly decreased in the ≥40-year-old group (18.8%) compared with that in the <35-year-old group (31.1%) and 35-39-year-old group (30.0%) (both P < 0.05). The live birth rate (30.6%, 21.7%, and 19.6%) was not significantly different across the paternal age subgroups (<35-year-old, 35-39-year-old, and ≥40-year-old, respectively; all P > 0.05), but showed a declining trend. The miscarriage rate significantly increased in the 35-39-year-old group (44.8%) compared with that in the <35-year-old group (21.0%; P < 0.05). No abnormality in newborn birth weight was found. The results indicated that paternal age over 40 years is a key risk factor that influences the assisted reproductive technology success rate even with good semen parameters, although it has no impact on embryo development.

Correction of the pathogenic mutation in the G6PC3 gene by adenine base editing in mutant embryos / 中华血液学杂志

Objective: To determine whether the adenine base editor (ABE7.10) can be used to fix harmful mutations in the human G6PC3 gene. Methods: To investigate the safety of base-edited embryos, off-target analysis by deep sequencing was used to examine the feasibility and editing efficiency of various sgRNA expression vectors. The human HEK293T mutation models and human embryos were also used to test the feasibility and editing efficiency of correction. Results: ①The G6PC3(C295T) mutant cell model was successfully created. ②In the G6PC3(C295T) mutant cell model, three distinct Re-sgRNAs were created and corrected, with base correction efficiency ranging from 8.79% to 19.56% . ③ ABE7.10 could successfully fix mutant bases in the human pathogenic embryo test; however, base editing events had also happened in other locations. ④ With the exception of one noncoding site, which had a high safety rate, deep sequencing analysis revealed that the detection of 32 probable off-target sites was <0.5% . Conclusion: This study proposes a new base correction strategy based on human pathogenic embryos; however, it also produces a certain nontarget site editing, which needs to be further analyzed on the PAM site or editor window.

Efeito do cilostazol na produção in vitro de ovinos / Effect of cilostazol on in vitro production of sheep / Efecto del cilostazol en la producción in vitro de ovejas

O objetivo deste estudo foi avaliar o efeito da inclusão de cilostazol no meio de maturação in vitro de oócitos sobre produção in vitro de embriões ovinos. Para isso, foram realizadas colheitas de oócitos oriundos de ovários obtidos em abatedouro por meio do método de aspiração folicular com bomba de vácuo. Os oócitos foram divididos em quatro grupos de maturação: grupo CON, onde os complexos cumulus oócitos foram imersos em TCM-199, suplementado com 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 µg de anfotericina, 0,2 mM de piruvato de sódio, 10% (v/v) de soro fetal bovino (SFB), 10 ng/mL de fator de crescimento epidérmico (EGF), 10 ug/m de FSH, 10 µg/mL de LH, 10 ug/mL de estradiol e 100 µM de cisteamina; e nos grupos CILO0,3; CILO1 e CILO10, os oócitos foram maturados no meio do grupo CON, mas sem a adição de cisteamina e suplementado com as concentrações de 0,3; 1 e 10 µM, respectivamente. Após 24h, os oócitos foram avaliados quanto a presença ou não de células do cumulus e quanto ao grau de expansão e destinados à fecundação in vitro, em meio FIV, juntamente com espermatozoides. Após a FIV, os presumíveis zigotos seguiram para o cultivo in vitro. Foram avaliadas clivagens no dia 2, sendo dia 0 o dia do início do CIV. Os resultados foram expressos em porcentagem e as variáveis de expansão das células do cumulus e número de estruturas clivadas foram comparadas por meio do teste qui-quadrado do software Epi Info (Epi Info 7.2.5, Atlanta, GA, EUA, 2021). Os resultados foram considerados significativos quando P<0,05. Em relação à expansão das células do cumulus, todos os grupos apresentaram 100% de expansão. Não houve diferenças significativas quanto ao grau de expansão das células do cumulus entre os grupos suplementados com cilostazol e cisteamina (P>0,05), assim como não houve diferenças significativas entre as taxas de clivagem entre os grupos suplementados com cilostazol e cisteamina (P > 0,05).

The objective of this study was to evaluate the effect of including cilostazol in the in vitro maturation medium of oocytes on the in vitro production of sheep embryos. Oocytes were collected from ovaries obtained from a slaughterhouse by follicular aspiration with a vacum pump. The oocytes were divided into four maturation groups: the CON group, where the cumulus-oocyte complexes were immersed in TCM-199 supplemented with 500 IU of penicillin, 0.5 mg of streptomycin, 1.25 µg of amphotericin, 0.2 mM of sodium pyruvate, 10% (v/v) fetal bovine serum (FBS), 10 ng/mL of epidermal growth factor (EGF), 10 µg/mL of FSH, 10 µg/mL of LH, 10 µg/mL of estradiol, and 100 µM of cysteamine; and in the CILO0.3, CILO1, and CILO10 groups, the oocytes were matured in the CON group medium without the addition of cysteamine and supplemented with concentrations of 0.3, 1, and 10 µM of cilostazol, respectively. After 24 hours, the oocytes were evaluated for the presence or absence of cumulus cells and the degree of expansion and then subjected to in vitro fertilization (IVF) with sperm in FIV medium. After IVF, the presumptive zygotes were cultured in vitro. Cleavage was evaluated on day 2, with day 0 being the start of IVF. Results were expressed as a percentage, and variables such as cumulus cell expansion and the number of cleaved structures were compared using the chi-square test in the Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. All groups showed 100% cumulus cell expansion, and there were no significant differences in cumulus cell expansion degree between the cilostazol- and cysteamine-supplemented groups (P > 0.05), as well as no significant differences in cleavage rates between the cilostazol- and cysteamine-supplemented groups (P > 0.05).

El objetivo de este estudio fue evaluar el efecto de la inclusión de cilostazol en el medio de maduración in vitro de ovocitos sobre la producción in vitro de embriones ovinos. Para ello, se realizaron recolecciones de ovocitos provenientes de ovarios obtenidos en un matadero mediante el método de aspiración folicular con bomba de vacío. Los ovocitos se dividieron em cuatro grupos de maduración: grupo CON, donde los complejos cúmulus ovocitos se sumergieron en TCM-199, suplementado con 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 ug de anfotericina, 0,2 mM de piruvato de sodio, 10% (v/v) de suero fetal bovino (SFB), 10 ng/mL de factor de crecimiento epidérmico (EGF), 10 ug/m de FSH, 10 µg/mL de LH, 10 µg/mL de estradiol y 100 µM de cisteamina; y en los grupos CILO0,3; CILO1 y CILO10, los ovocitos se maduraron en el medio del grupo CON, pero sin la adición de cisteamina y suplementado con las concentraciones de 0,3; 1 y 10 µM, respectivamente. Después de 24 horas, los ovocitos se evaluaron en cuanto a la presencia o no de células del cúmulus y em cuanto al grado de expansión y se destinaron a la fecundación in vitro, en medio FIV, junto con espermatozoides. Después de la FIV, los presuntos cigotos siguieron para el cultivo in vitro. Se evaluaron las clivajes en el día 2, siendo el día 0 el día del início del CIV. Los resultados se expresaron en porcentaje y las variables de expansión de las células del cúmulos y número de estructuras clivadas se compararon mediante la prueba del chi-cuadrado del software Epi Info (Epi Info 7.2.5, Atlanta, GA, EE. UU., 2021). Los resultados se consideraron significativos cuando P < 0,05. En relación a la expansión de las células del cúmulus, todos los grupos presentaron el 100% de expansión. No hubo diferencias significativas en cuanto al grado de expansión de las células del cúmulus entre los grupos suplementados con cilostazol y cisteamina (P > 0.05), así como no hubo diferencias significativas entre las tasas de clivaje entre los grupos suplementados con cilostazol y cisteamina (P>0,05).

El miocardio ventricular desde un enfoque cuantitativo. Su relación con la longitud del embrión humano / The ventricular myocardium from a quantitative approach. It ́s relation to the length of the human embryo

Introducción: Pese al incremento en conocimientos de la morfogénesis cardiaca humana, se conoce poco sobre los detalles cuantitativos en ello. Objetivo: Describir cuantitativamente el desarrollo del miocardio ventricular compacto y no compacto y su correlación con la longitud cráneo rabadilla. Métodos: Estudio descriptivo, transversal en 18 embriones humanos de los estadios 17 al 23 de Carnegie, pertenecientes a la Embrioteca de la Universidad de Ciencias Médicas de Villa Clara. Se midió la longitud del embrión, el grosor del miocardio compacto, trabecular y total en la pared lateral de ambos ventrículos y del vértice cardiaco. Resultados: El grosor de la pared lateral del miocardio compacto aumenta en ambos ventrículos desde los estadios 17 al 23 de Carnegie, de 0,06 mm hasta 0,17 mm en el derecho y de 0,09 mm hasta 0,23 mm en el izquierdo. El grosor de la pared lateral trabeculada disminuye con el avance de los estadios, de 0,43 mm a 0,34 mm en el derecho y de 0,45 mm a 0,37 mm en el izquierdo. El grosor de la pared lateral total aumenta de 0,48 mm a 0,51 mm en el ventrículo derecho y de 0,52 mm a 0,62 mm en el izquierdo. El grosor de la pared del vértice compacto aumenta de 0,19 mm a 0,25 mm. Conclusiones: La compactación de la pared ventricular aumenta con el desarrollo; la longitud cráneo raquis se relaciona con el grosor del miocardio ventricular(AU)

Introduction: Despite the increase in knowledge of human morphogenesis, especially cardiogenesis and the processes by which the morphology of the ventricular myocardium is defined, little is known about the quantitative details in it. Objectives: To quantitatively describe the development of compact and non-compact ventricular myocardium and its correlation with cranio-rump length. Methods: descriptive, cross-sectional study in 18 human embryos from Carnegie stages 17 to 23, belonging to the Embryoteca of the Villa Clara University of Medical Sciences. The length of the embryo, the thickness of the compact, trabecular and total myocardium were measured in the lateral wall of both ventricles and the cardiac apex. Results: The thickness of the lateral wall of the compact myocardium increases in both ventricles from Carnegie stages 17 to 23, from 0.06 mm to 0.17 mm in the right and from 0.09 to 0.23 mm in the left ventricles. The thickness of the trabeculated lateral wall decreases with the advancement of the stages, from 0.43 mm to 0.34 mm in the right and from 0.45 mm to 0.37 mm in the left. The total lateral wall thickness increases from 0.48 mm to 0.51 mm in the right ventricle and from 0.52 mm to 0.62 mm in the left. The wall thickness of the compact vertex increases from 0.19 mm to 0.25 mm. Conclusions: Ventricular wall compaction increases with development; the cranio-spinal length is related to the thickness of the ventricular myocardium(AU)

Gene editing of the human embryo: tensions and controversies among scientists / Edição genética do embrião humano: tensões e controvérsias de posicionamentos entre cientistas

Objective: This study analyzed the relation between the position of scientists on embryo editing and the different types of knowledge involved. Methods: A lexical analysis of 151 scientific articles in the PubMed and Web of Science databases was conducted using the IRAMUTEQ software. Results: The results showed that gene editing in embryos is presented in two argumentative branches: the first refers to the editing technique and its possibilities; the second discusses the impacts of these techniques on the public arena. The results demonstrate a consensus regarding the potential of editing; however, dilemmas about its effectiveness were also highlighted. Conclusion: The presence of ethical conflicts with embryo editing among the specialists was evidenced especially regarding the birth of genetically modified babies. Therefore, gene editing is marked by conflicts that are not limited only to biological contexts, but that encompasses different aspects of social life.

Objetivo: O objetivo deste trabalho foi analisar a relação entre o posicionamento dos cientistas sobre a edição de embriões e os diferentes tipos de conhecimento envolvidos nesses debates. Método: Utilizando o software IRAMUTEQ realizou-se uma análise lexical de 151 artigos científicos nas bases de dados PubMed e Web of Science. Resultados: Os resultados demonstraram que a edição genética de embriões se apresenta em dois blocos argumentativos: o primeiro se refere à técnica de edição e suas possibilidades e o segundo discute os impactos dessas técnicas na arena pública. Os achados demonstram consenso sobre as potencialidades da edição, contudo dilemas sobre a sua eficácia foram também destacados. Conclusão: Evidenciou-se a presença de embates éticos sobre a edição de embriões entre os especialistas em relação ao nascimento de bebês geneticamente modificados. Observou-se que a edição genética é marcada por conflitos que não se limitam apenas a contextos biológicos, mas que tangem diferentes aspectos da vida social.

Effect of Insemination Methods on Pregnancy Outcomes of Day 6 Blastocyst Transfer in Frozen Cycles / 中山大学学报(医学科学版)

ObjectiveTo compare the effects of two different insemination methods， conventional in vitro fertilization （IVF） and intracytoplasmic sperm injection （ICSI）， on pregnancy outcomes in patients with frozen-thawed D6（day 6） blastocyst transfer. MethodsA retrospective cohort study was conducted to analyze the clinical data of patients with thawed D6 blastocyst transfer between January 2018 and April 2020 at the Fertility Center of the Third Hospital of Sun Yat-sen University， divided into conventional IVF group （446 cycles ） and ICSI fertilization group （200 cycles） according to the fertilization method. Patients were divided into those with a history of D5（day 5） blastocyst transfer and those without. The patients’ general characteristics， blastocyst quality， and pregnancy outcomes of the two groups were compared. ResultsBMI， years of infertility， and basal FSH were not statistically significant in the IVF and ICSI groups （P > 0.05）. Regardless of the history of D5 transfer， patients in the ICSI group were younger than those in the IVF group （P < 0.001）， the proportion of primary infertility was significantly higher in the ICSI group （P < 0.001）， and the number of oocytes obtained and the number of normally fertilized oocytes in the ICSI group were higher than those in the conventional IVF fertilization group （P < 0.001）. The proportion of stage V and Ⅵ blastocysts was significantly higher in the conventional IVF group than in the ICSI group （21.6 % vs. 3.14 %， P < 0.001）. High-quality blastocysts with an ICM score of A were significantly higher in the ICSI group than in the IVF group （23.8 % vs. 14.3 %， P = 0.01）. The HCG-positive and clinical pregnancy rates were significantly higher in the ICSI group than in the IVF group （65.5 % vs. 48.4 %， P < 0.001； 56 % vs. 41.3 %， P = 0.001）， and embryo implantation and live birth rates were also higher in the ICSI group than in the conventional IVF group （43.8 % vs. 30.9 %， P < 0.001； 43.0 % vs. 31.8 %， P = 0.006）. After correcting for age and number of oocytes obtained between the two groups， the clinical pregnancy rate was still significantly higher in the ICSI group than in the conventional IVF group （OR： 1.590， 95 % CI： 1.030， 2.455， P = 0.036）. Infant birth weight was lower in the ICSI group than in the IVF group （P = 0.016）， and the differences in preterm birth rate， sex ratio， and mode of delivery were not statistically significant between the two groups. ConclusionsClinical pregnancy and live birth rates after thawing and transfer of D6 blastocysts fertilized by ICSI are higher than those of D6 blastocysts fertilized by conventional IVF， which may be related to the different factors contributing to the slow development of blastocysts in patients who received different fertilization methods. The relatively good pregnancy outcome after the transfer of thawed D6 blastocysts fertilized by ICSI may compensate to some extent for the difference in pregnancy outcome due to the relatively slow blastocyst development and a relatively higher proportion of D6 blastocysts after ICSI fertilization in male infertility patients.