Application research of PGT in blocking the inheritance of novel mutations in the PKHD1 gene in autoso-mal recessive polycystic kidney disease pedigrees / 实用医学杂志

Objective To investigate the application value of single nucleotide polymorphism(SNP)linkage analysis based on next-generation sequencing(NGS)technology in preimplantation genetic testing(PGT)of families with autosomal recessive polycystic kidney disease(ARPKD).Methods A family with ARPKD was selected,where the female member had a pregnancy ultrasound revealing polycystic kidney in the fetus.Genetic testing showed compound heterozygous mutations of the polycystic kidney/polycystic liver disease 1 gene(PKHD1),c.10444C>T(paternal)and c.4303del(maternal),with the c.4303del mutation being reported for the first time.Targeting the coding region of the PKHD1 gene,335 high-density tightly linked SNP sites were selected in the upstream and downstream 2M regions using multiplex polymerase chain reaction(PCR)and NGS.The couple′s SNP risk haplotypes carrying gene mutations were constructed.After in vitro fertilization,blastocyst culture was performed.Trophoblastic cells obtained from the biopsy were subjected to whole-genome amplification,and NGS was used for linkage analysis and low-depth chromosomal aneuploidy screening of the embryos.Sanger sequencing was used to verify the results of embryo linkage analysis.Results Among the 6 biopsied embryos,4 were mutation-free and euploid,1 exhibited heterozygous for the mutation and mosaic while another unstable sequencing data,making it impossible to judge.One of the mutation-free and developmentally healthy euploid embryos was implanted into the maternal uterus,resulting in the full-term delivery of a healthy baby.Conclusion Application of NGS-based SNP linkage analysis in PGT can effectively blocking the vertical transmission of ARPKD within families,while avoiding abortion issues caused by aneuploid embryos.This study is also the first PGT report target-ing the PKHD1 gene c.4303del mutation.

Application of Pre-implantation Genetic Testing in the Patients with Turner Syndrome / 昆明医科大学学报

Objective To investigate the application value of pre-implantation genetic testing(PGT)in patients with Turner syndrome.Methods The clinical data,embryonic development,PGT results and pregnancy outcome of 18 patients with Turner syndrome who underwent PGT in the reproductive center of 900th Hospital from January 2016 to June 2023 were retrospectively analyzed.Results All 18 patients had spontaneous puberty development,of which 4 patients had primary ovarian insufficiency(POI).A total of 24 oocyte retrieval cycles were performed in 18 patients,of which 6 patients had no biopsied embryos for 10 cycles.Sixty-one embryos were biopsied and 60 embryos were clearly diagnosed,including 25 with chromosomal abnormalities.Seven patients with mosaic Turner syndrome obtained clinical pregnancies after transplantation,including 4 healthy boys had already been delivered and 3 are in pregnancy.Conclusion There are numerous types of karyotype in Turner syndrome.The clinical phenotypes vary greatly in individuals with Turner syndrome,and prognosis of PGT is significant different.Patients with Turner syndrome who had biopsied embryos can obtain available embryo using PGT,and achieve ideal clinical outcomes.

Effects of different endometrial preparation regimens on pregnancy outcomes and obstetrical outcomes for first euploid single blastocyst transfer after PGT-A / 实用医学杂志

Objective To explore the effects of different endometrial preparation regimens on the pregnancy and obstetrical outcomes of single euploid blastocyst transfer after preimplantation genetic testing for aneuploidies(PGT-A).Methods The clinical data on patients with single euploid blastocyst transfer after PGT-A in our center from September 2015 to July 2021 were analyzed retrospectively.According to the different preparation regimens,the patients were divided into a natural cycle group(n = 80 cycles)and a hormone replacement cycle group(n = 259 cycles).The differences of pregnancy and obstetrical outcomes between the two groups were com-pared.Binary logistic regression was used to explore the influencing factors of pregnancy and obstetrical outcomes after PGT-A.The pregnancy outcomes of different endometrial preparation regimens were further explored by age stratification.Results As compared with that in the natural cycle group,the rate of high-quality blastocyst was significantly higher in the hormone replacement cycle group,but the live birth rate was lower and the abortion rate was higher(P<0.05).There was no statistical difference in the clinical pregnancy rate between the two groups.The incidence of cesarean section and macrosomia was higher in the hormone replacement cycle group than that in the natural cycle group,while the rate of low birth weight was lower,but there were no significant differences(P>0.05).Binary logistic regression showed that female age was an independent influencing factor of abortion.Female age and the days of blastocyst development were independent influencing factors of live birth.However,endometrial prepa-ration regimen was not a factor affecting pregnancy and obstetrical outcomes(P>0.05).The results of further analysis showed that the live birth rate in the natural cycle group was significantly higher than that in the hormone replacement cycle group,while the abortion rate in the natural cycle group was significantly lower than that in the hormone replacement cycle group for female patients aged over 35(P<0.05).Conclusions The endometrial preparation regimens did not affect the pregnancy and obstetrical outcomes of patients receiving single euploid blas-tocyst transfer after PGT-A.However,in the older patients,the natural cycle regimen can obtain a higher live birth rate and a lower abortion rate.

Factors correlating to frozen⁃thawed embryo transfer outcomes in preimplantation genetic testing cycles / 安徽医科大学学报

Objective@#To explore the factors associated with the transplant outcomes after the treatment of preimplantation genetic testing (PGT) for infertile patients.@*Methods@# A retrospective analysis of data of frozen⁃thawed embryo transfers in PGT for structural rearrangements/PGT for monogenic ( PGT⁃SR/PGT⁃M) cycles was performed. According to the transplant outcomes , it was divided into the pregnancy group (74 cases) and the nonpregnancy group (74 cases) . The factors including male age , female age , duration of infertility , number of abortion , female BMI , follicle⁃stimulating hormone (FSH) , luteinizing hormone ( LH) , estradiol( E2) , progesterone(P) , blood glucose , insulin , albumin , total cholesterol , triglycerides , high⁃density lipoprotein( HDL) , parents ′causes of infertility , type of chromosomal abnormality , number of retrieved oocyte , number of meiosis II( MII) , number of fertilization , number of two pronuclei(2PN) , endometrial thickness on embryo transfer day , endometrial preparation , day of embryo development , presence of a top quality embryo ( TQE) or not were analyzed by the Mann⁃Whitney U test and Pearson ′s chi⁃square test. The probability of transplant outcomes in correlation with the relevant parameters analyzed was measured with multivariate logistic regression analysis.@*Results@#Univariate analysis indicated that E2 , insulin , total cholesterol , triglycerides and endometrial preparation were significantly associated with transplant outcomes in PGT⁃SR , PGT⁃M cycles (P < 0. 05) . Multiple logistic regression analysis revealed that lower levels of insulin ( OR = 0. 14 , 95% CI = 0. 03 - 0. 27) and triglycerides ( OR = 1. 38 , 95% CI = 0. 14 -2. 78) in women were beneficial to transplant outcomes in PGT cycles. For endometrial preparation , compared with natural cycles , hormone replacement therapy (HRT) cycles ( OR = 3. 52 , 95% CI = 2. 54 - 4. 65) were another factor to improve transplant outcomes.@*Conclusion@#There is a correlation between the levels of insulin and triglycerides , endometrial preparation and transplant outcomes in PGT cycles. In detail , lower levels of insulin and triglycerides , and the HRT cycle can lead to a better transplant outcome.

Analysis of pregnancy outcome of preimplantation genetic testing for aneuploidy in cycles with different indications / 安徽医科大学学报

Objective @#To analyze the pregnancy outcome of preimplantation genetic testing for aneuploidy (PGT-A) in cycles with different indications．@*Methods @#The clinical information of 549 couples who underwent PGT-A were retrospectively analyzed．The cycles were divided into 6 groups according to the indication for PGT-A，namely : recurrent pregnancy loss group(n = 304) ，repeated implantation failure group(n = 57) ，advanced age group( ≥38 years old，n = 80) ，history of adverse pregnancy group(chorionic trisomy or adverse pregnancy，n = 24) ，male factor infertility group (n = 67) ，and abnormal sex chromosome number group (n = 17) ．The basic information，the number of retrieved oocytes，embryo biopsy result and pregnancy outcome were compared among different indication groups. @*Results@#The average age and days of gonadotropin ( Gn) used among the six groups were statistically different (P＜0. 001) ．The average number of retrieved oocytes，the rate of good-quality embryos，mosaic embryos， abnormal embryos and normal embryos，the average ovarian sensitivity index ( OSI) among the six groups were statistically different (P = 0. 03，P ＜0. 001，P = 0. 03，P ＜0. 001，P ＜0. 001，P ＜0. 001 ) ．Advanced age group had the highest rate of abnormal embryos，the least average number of retrieved oocytes，the lowest OSI and the lowest rate of normal embryos．There were statistical differences in clinical pregnancy rate，ongoing pregnancy rate and cumulative pregnancy rate per oocyte retrieved cycle (P＜0. 001) among the six groups，but there were no statistical differences in clinical pregnancy rate，ongoing pregnancy rate per transfer cycle and cumulative pregnancy rate among the five groups except for the male factor infertility group．@*Conclusion@#PGT-A can detect euploid embryo to transfer thereby improving pregnancy efficiency．The advanced age women have normal embryo to transfer and can obtain a better pregnancy rate，which may shorten their time of“take- baby -home ”．At the same time， PGT-A can significantly improve the pregnancy outcome of those with male factor infertility.

Application of Optical Genome Mapping Technology in Detecting Complex Chromosomal Rearrangement / 中山大学学报(医学科学版)

ObjectiveTo investigate the application of optical genome mapping （OGM） technology in detecting complex chromosomal rearrangement. MethodsWe recruited five patients who were diagnosed as complex chromosomal rearrangement at the Reproductive Medicine Center of the Sixth Affiliated Hospital of Sun Yat-sen University from January 2022 to June 2023. They underwent OGM， nanopore sequencing and pre-implantation genetic testing （PGT）. The results were compared with the results of karyotype and chromosomal microarray analysis （CMA）/ copy number variation sequencing （CNV-Seq）. ResultsOGM could detect translocation， invert inversion， and triplet translocation， which were consistent with the results of OGM and CMA/ CNV-Seq. But OGM could not detect Robertsonian translocation. ConclusionBecause of its ultra-long reads， OGM realizes the detection across repetitive regions， and it has great advantages when applied in patients with complex chromosome rearrangement or uncertain karyotype analysis. It can accurately locate breakpoints.

Preimplantation genetic testing and prenatal diagnosis in a family with type Ⅰ neurofibromatosis / 中华围产医学杂志

We report the implantation genetic testing and prenatal diagnosis of a family with neurofibromatosis type I (NF1). High-throughput sequencing combined with multiplex ligation-dependent probe amplification was performed to identify the pathogenic mutation sites, then verified by Sanger sequencing. The pathogenic mutation of c.4172G>C in the NF1 gene was found in the proband and his mother. After sequencing and single nucleotide polymorphism (SNP) haplotyping of the mutation sites in the embryos by establishing the SNP-linked haplotype, a well-developed blastocyst, without pathogenic mutations, was transplanted, and 28 d later, the ultrasound confirmed that the patient was pregnant. Amniotic fluid samples of the fetus were obtained at 19 +3 weeks for karyotyping and detection of the gene mutation site, which found the fetus did not carry the maternal c.4172G>C mutation of NF1 gene or any copy number variants of clear clinical significance. The patient delivered a healthy term girl by cesarean section, and no significant abnormalities were found during the follow-up to 10 months of age.

Application and development of molecular diagnostic techniques in human assisted reproduction / 中华检验医学杂志

In the past few years, with the breakthrough development of DNA sequencing technology, preimplantation genetic testing (PGT) has been widely used as an important part of assisted reproductive technology (ART). The progress of next generation sequencing (NGS) technology and the improvement of resolution, as well as the comprehensive application of molecular diagnostic technology makes it possible to perform the extensive and comprehensive chromosome screening and the clinical valuable detection of small gene fragment missing and repetition simultaneously, which is of great significance in terms of improving the pregnancy rate and reducing the abortion rate, multiplets rate and malformation rate. The common PGT molecular diagnostic techniques involves fluorescence in situ hybridization (FISH), single nucleotide polymorphism array(SNP-array), quantitative polymerase chain reaction(qPCR) and NGS. And each of them has its own highlights in clinical application and there are many uncertainties that are difficult to control. Moreover, ethical concerns brought about by technological progress also need to be addressed.

Application progress of single cell sequencing technology in diagnosis of genetic and reproductive / 国际生物医学工程杂志

The technique of prenatal diagnosis before embryo implantation has been greatly developed in assisted reproduction, especially for the people with abnormal genetic material to provide the technical possibility of eugenics and eugenics. As an emerging sequencing technology, single-cell sequencing can analyze the genome and transcriptome of cells from the level of a single cell, and reflect the heterogeneity between cells, thus helping to reveal the mechanism of the occurrence and development of diseases. Through prenatal diagnosis before embryo implantation and high-throughput sequencing of single cells obtained from embryo biopsy, euploidy of embryonic chromosomes can be effectively detected, and SNPs and chromosomal copy number variation, insertion, deletion and other variations can also be better detected. It can conduct the comprehensive detection and research of genomic polymorphism and mutation of individual and other species. In this paper, the single-cell sequencing technology and relevant methods was introduced, and the application scenarios of single-cell sequencing in genetic reproduction diagnosis were summarized. The applications of the technology in the field of genetic reproduction were described, and the future directions of the technology were discussed.

Impact of inactivated SARS-CoV-2 vaccination on embryo ploidy: a retrospective cohort study of 133 PGT-A cycles in China

BACKGROUND: Unsubstantiated concerns have been raised on the potential correlation between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination and infertility, leading to vaccine hesitancy in reproductive-aged population. Herein, we aim to evaluate the impact of inactivated SARS-CoV-2 vaccination on embryo ploidy, which is a critical indicator for embryo quality and pregnancy chance. METHODS: This was a retrospective cohort study of 133 patients who underwent preimplantation genetic testing for aneuploidy (PGT-A) cycles with next-generation sequencing technology from June 1st 2021 to March 17th 2022 at a tertiary-care medical center in China. Women fully vaccinated with two doses of Sinopharm or Sinovac inactivated vaccines (n = 66) were compared with unvaccinated women (n = 67). The primary outcome was the euploidy rate per cycle. Multivariate linear and logistic regression analyses were performed to adjust for potential confounders. RESULTS: The euploidy rate was similar between vaccinated and unvaccinated groups (23.2 ± 24.6% vs. 22.6 ± 25.9%, P = 0.768), with an adjusted ß of 0.01 (95% confidence interval [CI]: -0.08-0.10). After frozen-thawed single euploid blastocyst transfer, the two groups were also comparable in clinical pregnancy rate (75.0% vs. 60.0%, P = 0.289), with an adjusted odds ratio of 6.21 (95% CI: 0.76-50.88). No significant associations were observed between vaccination and cycle characteristics or other laboratory and pregnancy outcomes. CONCLUSIONS: Inactivated SARS-CoV-2 vaccination had no detrimental impact on embryo ploidy during in vitro fertilization treatment. Our finding provides further reassurance for vaccinated women who are planning to conceive. Future prospective cohort studies with larger datasets and longer follow-up are needed to confirm the conclusion.

Effectiveness of noninvasive embryo chromosome screening in preimplantation genetic testing at different ages / 中国医师杂志

Objective:To study the effectiveness of noninvasive embryo chromosome screening (NICS) based on blastocyst culture medium and cystic fluid in preimplantation genetic detection (PGT) of embryos in different age groups.Methods:A retrospective analysis of 62 couples who underwent PGT assisted pregnancy in Shenzhen Hospital of Peking University from January 2019 to June 2021. A total of 310 blastocysts were biopsied. At the same time, D3-6 blastocyst culture medium and blastocyst cavity fluid were collected for NICS. According to the age of the patients, they were divided into three groups: <35 years old group, 35≤age<40 years old group and ≥40 years old group. The results of NICS were compared with those of embryonic trophoblast (TE) biopsy in each group, and the consistency, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated. Then the consistency, sensitivity, specificity, positive predictive value and negative predictive value of NICS and TE among the three age groups were statistically analyzed.Results:There was no statistically significant difference in the consistency rate of NICS and TE among the three age groups ( P>0.05), but there was an upward trend in the elderly group (35≤age<40 years and ≥40 years). There was no statistically significant difference in specificity, sensitivity and PPV among the three age groups ( P>0.05). There was significant difference in NPV between the ≥40 years group and the other two groups ( P<0.05). Conclusions:There was no statistical difference in the effectiveness of NICS among different age groups, However, there was an increasing trend in people ≥35 years of age.

Predictive factors for the successful implantation and live birth after Euploid Blastocyst Transfer: A single center study

Background@#Pre-implantation Genetic Testing for Aneuploidy (PGT-A) has enabled IVF specialists to screen embryos for abnormalities in chromosome number and structure. Subsequently, healthy embryos are selected for transfer, decreasing the rate of spontaneous miscarriages and improving pregnancy outcomes. In spite of this, almost only half of the PGTdetermined euploid embryos result in a pregnancy. @*Objective@#This study aimed to determine what other factors among euploid embryo transfers will have an association with successful implantation and live birth. @*Methods@#This study retrospectively analyzed 159 IVF-PGS cycles performed in CARMI SLMCGC from January 2017 to December 2019. Of these, a total of 231 euploid embryos (86 single embryo transfers and 73 double embryo transfers) were assessed. The relationship of eight predictive variables (maternal age, maternal BMI, etiology of infertility, history of failed IVF, blastocyst expansion stage, ICM grade, TE grade and endometrial thickness on transfer) with regard to the outcome of successful implantation and live birth among single or double euploid blastocyst transfers were determined via logistic regression analysis. @*Results@#Overall, the implantation rate was significantly lower when using B-grade ICM or C-grade ICM blastocysts as compared to A-grade ICM blastocysts (OR 0.54, 95% CI 0.356- 0.815, p = 0.003). With regard to live birth rate, the success of transfer is statistically lower when using a B-grade or C-grade ICM blastocysts as compared to A-grade ICM blastocysts (OR 0.55, CI 0.354-0.863, p = 0.009). Other predictive factors such as maternal age, maternal BMI, etiology of infertility, number of previous IVF, blastocyst expansion stage, trophectoderm grade and endometrial thickness had no apparent effect on the outcome of implantation and live birth. @*Conclusion@#Present study results suggest that only the ICM grade of euploid blastocysts correlates with implantation and live birth in IVF-FET cycles. Therefore, the selection of euploid blastocysts based on the presence of a higher grade ICM is the most predictive factor that determines success among those undergoing IVF with PGT-A.

PGT or ICSI? The impression of NGS-based PGT outcomes in nonmosaic Klinefelter syndrome / 亚洲男科学杂志(英文版)

This retrospective study demonstrates the clinical outcomes of patients with nonmosaic Klinefelter's syndrome (KS) who underwent preimplantation genetic testing (PGT) with frozen-thawed testicular spermatozoa. Microdissection testicular sperm extraction (micro-TESE) was performed for sperm retrieval. Next-generation sequencing (NGS) was conducted for embryo analysis. A total of 18 couples aged ≤35 years were included, and 22 oocyte retrieval cycles were completed. Euploidy was detected in 29 of 45 (64.4%) embryos. Additionally, the numbers of aneuploid and mosaic embryos detected were 8 (17.8%) and 8 (17.8%), respectively, regardless of a lack of sex chromosome abnormalities. Finally, 13 couples with euploid embryos completed 14 frozen embryo transfer (FET) cycles. Ten couples had clinical pregnancies, and 6 of them had already delivered 5 healthy babies and 1 monozygotic twin. There were also 4 ongoing pregnancies and 2 biochemical pregnancies, but no early pregnancy loss was reported. Based on our results, we speculate that for KS patients, when sperm can be obtained by micro-TESE, the cryopreservation strategy makes the ovarian stimulation procedure more favorable for female partners. The paternal genetic risk of sex chromosome abnormalities in their offspring is extremely low in men with KS. In addition to PGT, the intracytoplasmic sperm injection (ICSI) procedure is comparably effective but more economical for young nonmosaic KS couples. ICSI should be offered as an option for such couples, but monitoring by prenatal genetic diagnosis is recommended.

Técnicas de substituição de DNA mitocondrial para reduzir a transmissão da Síndrome de Leigh / Mitochondrial DNA replacement techniques to reduce Leigh syndrome transmission

A Síndrome de Leigh (SL) é uma doença neuro-metabólica congênita, que faz parte do grupo das encefalopatias fatais, com progressão e morte dentro de 2 anos, em média. A SL é causada por mutações no DNA que causam alterações na geração de ATP celular pelas mitocôndrias. As mitocôndrias contêm seu próprio DNA (mtDNA) e, ao contrário do DNA nuclear, o mtDNA é herdado somente da mãe. Mulheres portadores de mutações causadoras da SL podem vivenciar experiências muito tristes ao tentarem realizar o sonho da maternidade. As técnicas de substituição de mtDNA mutado com mtDNA saudável de doadora, oferecem a essas mulheres a possibilidade de terem uma criança geneticamente relacionada sem a SL. O desenvolvimento e a aplicação clínica de terapias de substituição de mtDNA já são uma realidade, tendo o primeiro bebê gerado a partir da técnica nascido em 2016. Mas será que essas técnicas são seguras? Neste trabalho, revisamos a SL e algumas técnicas de substituição de mtDNA já aplicadas em humanos, que envolvem a transferência de pronúcleos de zigotos ou de fuso acromático de oócitos. Concluímos que, apesar dos resultados promissores, ainda é cedo para assegurar a aplicabilidade clínica de técnicas de substituição de mtDNA em seres humanos. (AU)

Leigh syndrome (SL) is a congenital neurometabolic disease included in the group of fatal encephalopathies, with progression and death within 2 years on average. SL is caused by mutations in the DNA that cause changes in the generation of cellular ATP by mitochondria. Mitochondria contain their own DNA (mtDNA) and, unlike nuclear DNA, mtDNA is inherited only from the mother. Women with SL mutations may experience mournful situations when attempting to fulfill the dream of motherhood. Techniques for replacing mutant mtDNA with healthy donor mtDNA provide these women with the possibility of having a genetically related child without SL. The development and clinical application of mtDNA replacement therapies is a reality, and the first baby generated using the technique was born in 2016. However, are these techniques safe? In this article, we review SL and some mtDNA replacement techniques that have been used in humans, which involve zygote pronuclear transfer or oocyte spindle transfer. We conclude that, despite the promising results, it is too early to ensure that mtDNA replacement techniques are clinically applicable to humans. (AU)

Birth of healthy neonate following preimplantation genetic diagnosis in a mother with mosaic Turner syndrome. Case report and review of the literature / Recién nacido sano después de diagnóstico genético preimplantatorio en una madre con síndrome de turner mosaico. reporte de caso y revisión de la literatura

RESUMEN Objetivos: reportar el caso de una paciente con síndrome de Turner en mosaico, a quien se le realizó un tratamiento de reproducción asistida con análisis genético preimplantatorio para aneuploidias, logrando el nacimiento de una niña sana con cariotipo normal, y realizar una revisión de la literatura sobre la utilidad del diagnóstico genético preimplantatorio en las mujeres con síndrome de Turner. Materiales y métodos: se presenta el caso de una mujer de 27 años, con diagnóstico de síndrome de Turner en mosaico y con alteración secundaria en la reserva ovárica, atendida en centro de referencia para el manejo de infertilidad en Medellín, Colombia, a quien se le realizó un tratamiento de fertilización in vitro con análisis genético preimplan-tatorio para prevenir la transmisión del síndrome de Turner a su descendencia. Se realizó una búsqueda de la literatura en las bases de datos Medline vía PubMed, Clinical Key, OVID, Embase, Lilacs, SciE- LO y Oxford Journals, con los siguientes términos: "Turner Syndrome", "Mosaic Turner", "Preim- plantation Genetic Screening", "Preimplantation Genetic Testing", "Preimplantation Genetic Diagnosis", "Pregnancy", "Successful pregnancy". Como criterios de inclusión se consideraron artículos tipo series y reportes de casos, cohortes y artículos de revisión desde enero de 1980 hasta junio de 2017, que incluyeran mujeres con síndrome de Turner embarazadas por medio de técnicas de fertilización in vitro, con sus propios óvulos, y que hubiesen sido sometidas a biopsia embrionaria para diagnóstico genético preimplantatorio. La búsqueda se limitó a los idiomas español e inglés. Resultados: un estudio cumplió con los criterios de inclusión. Tanto en este reporte como en nuestro caso, las pacientes con síndrome de Turner en mosaico se sometieron a varios ciclos de inyección intracitoplasmática de espermatozoides (ICSI) con sus propios óvulos, luego se realizó biopsia em- brionaria para análisis genético preimplantatorio utilizando diferentes técnicas. En ambos casos se logró la transferencia al útero de embriones euploides con el posterior nacimiento de niñas sanas con cariotipo normal. Conclusión: Las pacientes con ST mosaico podrían beneficiarse de la biopsia embrionaria y análisis genético preimplantatorio para prevenir la transmisión del defecto genético a su descendencia.

ABSTRACT Objectives: To report the case of a patient with mosaic Turner syndrome who underwent assisted reproduction treatment with preimplantation genetic testing for aneuploidy and gave birth to a healthy baby girl with normal karyotype; and to conduct a review of the literature on the usefulness of preimplantation genetic diagnosis in women with Turner syndrome. Materials and methods: A case of a 27 year-old woman diagnosed with mosaic Turner syndrome and secondary altered ovarian reserve, seen in a referral center for infertility management in Medellín, Colombia. The patient underwent in vitro fertilization followed by pre-implantation genetic testing to prevent transmission of Turner syndrome to her progeny. A literature search was conducted in the Medline via PubMed, Clinical Key, OVID, Embase, Lilacs, SciELO and Oxford Journals data- bases using the following terms: "Turner Syndrome," "Mosaic Turner," "Preimplantation Genetic Screening," "Preimplantation Genetic Testing," "Preimplantation Genetic Diagnosis," "Pregnancy," "Successful pregnancy." Inclusion criteria were case series and case reports, cohort studies and review articles published between January 1980 and June 2017 that included women with Turner syndrome achieving pregnancy by means of in vitro fertilization techniques with their own oocytes and who had undergone embryo biopsy for preimplantation genetic diagnosis. The search was limited to articles in Spanish and English. Results: one study met the inclusion criteria. Both in this report and in our case, patients with mosaic Turner syndrome underwent several cycles of intracytoplasmic sperm injection (ICSI) with their own eggs, then performed embryonic biopsy for preimplantation genetic analysis using different techniques. In both cases, euploid embryos were transferred to the uterus with the subsequent birth of healthy girls with normal karyotype. Conclusion: Patients with mosaic Turner syndrome could benefit from preimplantation biopsy and genetic analysis to prevent transmission of the genetic defect to their progeny.

Diagnóstico genético preimplantacional no invasivo; datos preliminares en México / Non-invasive preimplantation genetic diagnosis; preliminary data in Mexico

Resumen: OBJETIVO: Analizar las tasas de concordancia, falsos positivos y negativos entre el ADN embrionario circulante en medio de cultivo y su relación con los reportes de la biopsia de trofoectodermo. MATERIALES Y MÉTODOS: Estudio observacional, prospectivo y comparativo, llevado a cabo en el Centro de Reproducción Arcos Nascere en noviembre 2018. Criterios: de inclusión: parejas en esquema de fertilización in vitro, con diagnóstico genético preimplantacional de aneuploidias. Criterios de exclusión: pacientes con anomalías estructurales o enfermedades monogénicas. Criterio de eliminación: blastocistos con eclosión asistida. Variables de respuesta: tasa de concordancia, falsos positivos y negativos entre las biopsias de trofoectodermo y los medios de cultivo. El análisis estadístico se realizó con SPSS 25.0, con pruebas t de Student y χ2 con valor de p < 0.05 significativa. RESULTADOS: Se analizaron 20 blastocistos de 5 parejas y se obtuvieron resultados informativos de 17 (amplificación global exitosa); 70% en día 5 y 100% en día 6. La tasa general de concordancia entre las biopsias de trofoectodermo y los medios de cultivo fue de 68.7% (42.8% en día 5 y 88.8% en día 6). En cuanto a las discrepancias, solo se observaron 2 falsos negativos en los medios de cultivo vs la biopsia de trofoectodermo (14.2% en día 5 y 11.11% en día 6); hubo 3 casos de falsos positivos (la mitad en día 5 y ninguno en día 6-7). CONCLUSIONES: Con la prueba genética no invasiva de aneuploidias se alcanzaron altas tasas de concordancia, sobre todo en embriones en día 6.

Abstract: OBJECTIVE: Analyze the concordance, false positive and false negative rates between circulating free DNA of the culture media compared to the results of the trophectoderm biopsy. MATERIALS AND METHODS: Observational, prospective and comparative study, conducted at Arcos Reproduction Center S.C. Nascere in november 2018. Couples with indication of preimplantation genetic diagnosis of aneuploidies undergoing In vitro Fertilization were included; carriers of structural anomalies or monogenic diseases were excluded and blastocysts with assisted hatching were eliminated. The response variables were the concordance, false positives and false negatives rates between trophoctoctoderm biopsies and culture media. Statistical analysis was performed with SPSS 25.0, using t-Student and chi-square tests with a value of p <0.05 significant. RESULTS: Informative results were obtained in 17 of the 20 culture media (85% successful global amplification); 70% on day 5 and 100% on day 6. The general concordance rate between trophectoderm biopsies and culture media was 68.7% (42.8% on day 5 and 88.8% on day 6). Regarding discrepancies, only 2 false negatives were observed in the culture media compared to the trophectoderm biopsy (14.2% on day 5 and 11.1% on day 6). There were 3 cases false positives (42.8% on day 5 and 0% on day 6). CONCLUSIONS: High rates of concordance were reached with the non-invasive genetic aneuploidy test, mainly in embryos on day 6.

Case reports for embryo banking: for women who want their own

Out of the many challenges in management of female factor infertility, poor responders and low response to stimulation in aged and even younger women, seems to be a common problem. It is very difficult to offer one particular management strategy or treatment protocol for optimum outcome in this group of women of poor responders. In a low resource set up, IVF (In vitro Fertilization) specialist doctors usually face a challenge in treating women with poor/ low ovarian reserve as ovum / gamete donation is considered as a taboo in various sections of society even today. Hence women insist on having an offspring of "their own" and vehemently deny ovum / gamete donations. In this article we discuss 2 cases of poor ovarian reserve retrospectively, who underwent multiple cycles of controlled ovarian hyperstimulation for embryo banking and ultimately achieved pregnancy. Both patients achieved pregnancy with the method of embryo banking. Embryo banking should be considered and discussed. Various articles have discussed the advantages and disadvantages of embryo banking or even oocytes accumulation. The advantages of this technique is patients with poor/low ovarian reserve get a chance to be pregnant with their own oocytes and also have a chance for vitrification of residual embryos. Another advantage in such patients is that the embryos can undergo PGS (Preimplantation Genetic Screening) techniques in cases of suspected genetic disorders. The disadvantage in a low resource set up like India is the cost of the treatment. Nevertheless, embryo banking and accumulation of oocytes should be given as an option for treatment of poor/ low ovarian reserve and could be considered as a ray of hope for all future mothers hoping for a child of "their own".

La importancia del asesoramiento genético para el diagnóstico genético preimplantacional de aneuploidías (PGT-A)

The development of new genomic technologies has strengthened the influence of genetics in all medical specialties; reproductive medicine is no exception. The introduction of new genetic tests to daily clinical practice, together with the complexity of genetic information and its potential psychological burden, make specialized genetic counseling essential. Preimplantation genetic testing for aneuploidies (PGT-A) has become an almost routine procedure in assisted reproduction treatments. Nevertheless, in Peru, patients usually receive none or inadequate corresponding genetic counseling, which hinders informed decision-making.

El desarrollo de las nuevas tecnologías genómicas ha potenciado la influencia de la genética en todos los campos de la medicina, en donde la medicina reproductiva no es la excepción. La frecuente introducción de nuevas pruebas genéticas en la práctica clínica diaria, junto con la complejidad de la información genética y su potencial carga psicológica, hacen indispensable el asesoramiento genético especializado. En este contexto, el diagnóstico genético preimplantacional para aneuploidías (PGT-A) se ha convertido en un procedimiento casi de rutina en los tratamientos de reproducción asistida. Sin embargo, en el Perú, en la gran mayoría de casos los pacientes no reciben el asesoramiento genético correspondiente o este no es el adecuado, que no permite una toma de decisiones informada.

El uso de la tecnología Time-Lapse y algoritmo predictivo KIDScore 5 ayuda a mejorar la selección de embriones euploides para la transferencia

Objectives: To determine if the use of the KIDScore 5 algorithm (known implantation data) can help select between euploid embryos in order to improve pregnancy and implantation rates in patients undergoing assisted reproductive procedures. Methods: Retrospective cohort study in a fertility clinic, from October 2016 to December 2018, of 1 049 embryos from 328 patients. All the embryos were cultured in the Time-Lapse, Embryoscope® incubator (Vitrolife®, Canada) for 5-6 days. Of these, 896 embryos (85.4%) were biopsied and analyzed by NGS, and assessed with the predictive KIDScore 5 algorithm (Vitrolife®, Canada). The 153 remaining embryos (14.6%) were assessed with the predictive KIDScore 5 algorithm only. 256 single euploid embryos were transferred in couples undergoing IVF treatments at the Inmater clinics laboratory of assisted reproduction in Lima - Peru. Results: The implantation rate was significantly higher (p = 0.004) in euploid embryos transferred when selected by the KIDScore 5 algorithm (Group 2) versus those selected using only genetic study by NGS technology (Group 1) (71.2% vs. 48.8%). The rate of implantation of the euploid embryos transferred with KIDScore value = 6 versus those transferred with KIDScore = 1 was statistically different (73.5% vs. 50.8%; p = 0.030). When assessing the relationship between the rate of euploid embryos versus the result of the KIDScore 5 value, we found highly significant differences in the rates of euploid embryos with values 6 and 5 versus those with KIDScore 0 and 1 (60.5% vs. 45.7%; p = 0.0004). Conclusions: The embryo selection with the KIDScore 5 algorithm offers advantage on implantation and pregnancy rates only when euploid embryos are transferred. Its use as an additional criterion to embryo selection should be considered when accompanied by genetic study of the embryos to be transferred. Euploid embryos with a higher value in the KIDScore 5 algorithm scale have better rates of implantation and euploidy than embryos with the minimum value of this algorithm.

Objetivos. Evaluar si el uso del algoritmo KIDScore 5 (known implantation data) puede ayudar a seleccionar entre los embriones euploides, para mejorar las tasas de embarazo e implantación en pacientes sometidas a procedimientos de reproducción asistida. Métodos. Estudio de cohorte retrospectivo en una clínica de fertilidad, desde octubre 2016 a diciembre 2018. Se estudió 1 049 embriones provenientes de 328 pacientes. Todos los embriones fueron cultivados en la incubadora Time-Lapse, Embryoscope® (Vitrolife®, Canadá) durante 5 a 6 días. De estos, 896 embriones (85,4%) fueron biopsiados y analizados mediante NGS y recibieron una valoración otorgada por el algoritmo predictivo KIDScore 5 (Vitrolife®, Canadá). Los 153 embriones restantes (14,6%) únicamente recibieron la valoración mediante el algoritmo predictivo KIDScore 5. Se realizó 256 transferencias únicas de embriones euploides en parejas sometidas a tratamientos de FIV en el laboratorio de reproducción asistida de la Clínica Inmater, Lima - Perú. Resultados. La tasa de implantación de los embriones euploides transferidos con valores de KIDScore = 6 versus los transferidos con valores de KIDScore = 1 tuvo diferencia estadísticamente significativa (73,5% vs. 50,8%; p=0,030). Al evaluar la relación entre la tasa de euploidia embrionaria versus el resultado del valor de KIDScore 5, se obtuvo diferencias altamente significativas en las tasas de euploidia en los embriones con resultados de KIDScore 6 y 5 versus los de KIDScore 0 y 1 (60,5% vs. 45,7%; p=0,0004). Conclusiones. La selección embrionaria con ayuda del algoritmo KIDScore 5 ofrece ventaja en las tasas de implantación y embarazo únicamente cuando se transfieren embriones euploides. Su uso como criterio adicional a la selección embrionaria debiera ser considerado siempre que se acompañe estudio genético a los embriones a transferir. Los embriones euploides con valor más alto en la escala del algoritmo KIDScore 5, tienen mejores tasas de implantación y euploidía que los embriones con el valor mínimo de dicho algoritmo.

Mechanism of male reproductive failure in autosomal dominant polycystic kidney disease and outcomes of assisted reproductive technology treatment / 上海交通大学学报（医学版）

Objective: To explore the potential mechanism of male reproductive failure in autosomal dominant polycystic kidney disease (ADPKD) patients and analyze the outcomes of assisted reproductive technology treatment. Methods: Next-generation sequencing was performed for genetic diagnosis of 8 ADPKD patients, who came to International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine, for genetic counseling. The semen of ADPKD patients and normal males who came for pre-pregnancy consultation was collected by masturbation for sperm analysis. The ultrastructure of sperm was observed by transmission electron microscopy. Outcomes of 7 patients with ADPKD who chose preimplantation genetic testing (PGT) were compared with those of 7 patients who were dystrophin (DMD) gene mutation carriers, undergoing the PGT in the same period. Results: Eight patients with ADPKD were heterozygous for polycystin 1 (PKD1) gene. Key parameters of sperm motion including progressive motility sperm percentage, curvilinear velocity, straight-line velocity, average path velocity, amplitude of lateral head displacement were much lower than those of normal semen, showing mild to severe oligozoospermia. One ADPKD patient with severe oligoathenospermia manifested bilateral seminal vesicle cysts. Transmission electron microscopy showed that the central microtubules of the sperm flagella of ADPKD patients were absent and the surrounding double microtubules were disorganized. There was no significant difference in the number of eggs, fertilization rate, cleavage rate, effective embryo rate and excellent embryo rate between the ADPKD patients and the DMD gene mutation carriers, but the ADPKD patients were prone to early abortion. Conclusion: Male reproductive failure caused by ADPKD may be related to many factors such as abnormal structure of sperm flagella and genital cysts. Further, PKD1 mutation may play a role in embryo implantation and early development.