Endometriosis, Oocyte, and Embryo Quality.

Endometriosis is a common finding among women with infertility, and women who are diagnosed with endometriosis are almost twice as likely to experience infertility. Mechanisms by which endometriosis causes infertility remain poorly understood. In this review, we evaluate the current literature on the impact of endometriosis on oocyte and embryo quality. The presence of endometriosis evidently reduces ovarian reserve, oocyte quality, and embryo quality; however, this does not appear to translate to a clear clinical impact. Analysis of data from large assisted reproduction technology registries has shown that women with endometriosis have a lower oocyte yield but no reduction in reproductive outcomes. There is a need for future studies in the form of well-designed randomized controlled trials to further evaluate the role of surgical and medical treatment options in women with endometriosis undergoing assisted conception.

Physiological intracytoplasmic sperm injection does not improve the quality of embryos: A cross-sectional investigation on sibling oocytes.

OBJECTIVE: This study aimed to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) and intracytoplasmic sperm injection (ICSI) in terms of the fertilization rate and embryo quality using sibling oocyte cycles. METHODS: This prospective, cross-sectional study collected data from 76 couples who underwent their first cycle at the Hue Center for Reproductive Endocrinology and Infertility, Vietnam, between May 2019 and November 2021. The inclusion criteria were cycles with at least eight oocytes and a sperm concentration of 5×106/mL. Sperm parameters, sperm DNA fragmentation (SDF), fertilization, and the quality of cleavage-stage embryos on day 2 and blastocysts on day 5 were examined. RESULTS: From 76 ICSI cycles, 1,196 metaphase II (MII) oocytes were retrieved, half of which were randomly allocated to either the PICSI (n=592) or ICSI (n=604) treatment group. The results showed no significant difference between the two groups in terms of fertilization (72.80% vs. 75.33%, p=0.32), day 2 cleavage rate (95.13% vs. 96.04%, p=0.51), blastulation rate (52.68% vs. 57.89%), and high-quality blastocyst rate (26.10% vs. 31.13%, p=0.13). However, in cases where SDF was low, 59 cycles consisting of 913 MII oocytes produced a considerably higher blastulation rate with PICSI than with ICSI (50.49% vs. 35.65%, p=0.00). There were no significant differences between the pregnancy outcomes of the PICSI and ICSI embryo groups following embryo transfer. CONCLUSION: Using variable sperm quality provided no benefit for PICSI versus ICSI in terms of embryo outcomes. When SDF is low, PICSI appears to be able to produce more blastocysts.

Patent hepatic ciliated foregut remnant resulting in an umbilicobiliary sinus tract, with gallbladder agenesis, in an 8-wk-old male French Bulldog.

Hepatic ciliated foregut remnants or cysts are congenital abnormalities resulting from retention of embryonic ciliated foregut within the liver. These structures are rarely reported in the human medical literature and have not been reported in the veterinary literature previously, to our knowledge. We describe here a case of an 8-wk-old male French Bulldog with a congenital patent hepatic ciliated foregut remnant resulting in an umbilicobiliary sinus tract. The dog also had concurrent gallbladder agenesis. The patient had yellow fluid discharging from the umbilicus, mimicking a patent urachus. Surgical exploration, removal, and histology provided a conclusive diagnosis of a hepatic foregut remnant and therapeutic resolution of the clinical signs. The histologic appearance of a hepatic foregut remnant is classical, namely a duct composed of 4 layers: an inner ciliated epithelial lining, loose connective tissue, smooth muscle, and a fibrous capsule.

Challenges in the use of nanostructures as carriers of nucleic acids in clinical practice

ABSTRACT The delivery of nucleic acids to cells is considered a crucial step for the success of genetic modifications aimed at therapeutic purposes or production of genetically modified animals. In this context, nanotechnology is one of the most promising fields of science, with the potential to solve several existing problems. Nanostructures have desirable characteristics to be used as carriers, such as nanometric size, large surface area, cell internalization capacity, prolonged and controlled release, among others. Genetically modified animals can contribute to the production of biopharmaceuticals, through the expression of high-associated-value molecules. The production of these animals, also known as biofactories, further enhances Brazilian agribusiness, since it allows adding value to the final product, and favors the integration between the agricultural market and the pharmaceutical sector. However, there is a growing concern about the safety and possible harmful effects of nanostructures, since data on the safe use of these materials are still insufficient. The objective of this review was to address aspects of the use of nanostructures, mainly carbon nanotubes as nucleic acid carriers, aiming at the production of genetically modified animals, with the certainty that progress in this field of knowledge depends on more information on the mechanisms of interaction between nanostructures, cells and embryos, as well as on its toxicity.

Fli1 Promotes Vascular Morphogenesis by Regulating Endothelial Potential of Multipotent Myogenic Progenitors.

[Figure: see text].

Embryonic ontogeny of three species of Horned Frogs, with a review of early development in Ceratophryidae.

Horned Frogs of the family Ceratophryidae are conspicuous anurans represented by three endemic South American genera. Most ceratophryids inhabit semiarid environments, but three species of Ceratophrys occupy tropical or temperate humid areas. Several morphological and behavioral characters of larvae and adults are conserved across the family. Based on examination of specimens and accounts in the literature, the embryonic development of C. ornata, C. cranwelli, and the monotypic genus Chacophrys are described and compared with that of species of Lepidobatrachus. Ceratophryid embryos share a suite of morphological features and heterochronic shifts during development. Most features, such as gill structure, ciliation, early hatching, and precocious differentiation of the gut and hind limbs, are shared by all the species regardless the differences in the habitats that occupy. This is consistent with previous observations of some adult characters, and likely supports the hypothesis of an early diversification of ceratophryids in semiarid environments. Other embryonic features, such as the morphology and ontogeny of the oral disc and digestive tract, are correlated with larval feeding habits and vary within the family. The evolutionary and ecological significance of some conserved characters (e.g., gastrulation pattern, Type-A adhesive glands) and other taxon-specific features (e.g., nasal appendix) remain to be explored in the group.

Desenlaces en ciclos de transferencia con embriones vitrificados: análisis de dos esquemas de preparación endometrial / Results in cycles of frozen-thawed embryo transfer: Analysis of two protocols of endometrial preparation

Resumen: OBJETIVO: Comparar la tasa de recién nacido vivo con la de embarazo clínico-transferencia embrionaria, las características clínicas y concentraciones hormonales entre dos esquemas de preparación endometrial para transferencia de embriones desvitrificados: con un agonista de GnRH versus su sustitución con dosis altas de estrógenos. MATERIALES Y MÉTODOS: Estudio observacional, de cohorte histórica, efectuado en pacientes atendidas en el Instituto Nacional de Perinatología en protocolo FIV-ICSI entre enero 2017-marzo 2019. Se compararon dos esquemas de preparación endometrial: grupo A con agonista de GnRH y grupo B con estradiol a dosis de 8 mg al día sin agonista de GnRH. A todas las pacientes se les tomaron muestras de sangre para determinación de FSH, LH, estradiol y progesterona y se dio seguimiento ultrasonográfico durante la preparación endometrial. RESULTADOS: En 99 pacientes entre 23 y 42 años, con embriones congelados, la tasa de recién nacido vivo-transferencia embrionaria fue, respectivamente, de 17.2 y 8.6% (OR 1.98; IC95%: 077-1.53) y la de embarazo clínico de 26.5 y 22.8% (OR1.09; IC95%: 0.77-1.53) en los grupos A y B, respectivamente. La duración total del ciclo, los días con estrógenos y la concentración sérica de estradiol al inicio de la progesterona alcanzaron diferencia significativa entre los grupos. No se detectaron datos clínicos ni de laboratorio sugerentes de ovulación. CONCLUSIONES: La tasa de recién nacido vivo fue mayor con agonistas de GnRH sin alcanzar significación; sin embargo, la preparación endometrial solo con estrógenos es un esquema seguro, de menor costo y más amigable para la paciente y el médico.

Abstract: OBJECTIVE: To compare the rate of live newborn with clinical pregnancy-embryo transfer, clinical characteristics and hormonal concentrations between two endometrial preparation schemes for devitrified embryo transfer: with a GnRH agonist (aGnRH) versus its replacement with high doses of estrogens. MATERIALS AND METHODS: Historical cohort observational study conducted in patients treated at the National Institute of Perinatology under IVF-ICSI protocol between January 2017-March 2019. Two endometrial preparation schemes were compared: group A with GnRH agonist and group B with estradiol at 8 mg per day without aGnRH. All patients had blood samples taken for FSH, LH, estradiol and progesterone determination and ultrasonographic follow-up during endometrial preparation. RESULTS: in 99 patients between 23 and 42 years of age, with frozen embryos, the live birth-embryo transfer rate was 17.2 and 8.6%, (OR 1.98; CI95%: 077-1.53) and the clinical pregnancy rate was 26.5 and 22.8% (OR1.09; CI95%: 0.77-1.53) in groups a and b, respectively. total cycle length, estrogen days and serum estradiol concentration at the beginning of progesterone reached significant difference between groups. no clinical or laboratory data suggesting ovulation were detected. CONCLUSIONS: The rate of live newborn was higher with GnRH agonists without reaching significance; however, endometrial preparation with estrogens alone is a safe, lower cost and more patient- and physician-friendLy scheme.

[The senses on in vitro embryos and the religious dimension: contributions for debate.] / Los sentidos sobre los embriones in vitro y la dimensión religiosa: aportes para el debate.

Based on a biographical investigation of the experiences of assisted reproduction users in Córdoba, this work seeks contribute on the religious dimension as a category that a priori does not determine the users senses about reproduction and embryos in vitro.

A partir de una investigación biográfica sobre experiencias de usuarias de reproducción asistida en Córdoba, este trabajo busca realizar un aporte sobre la dimensión religiosa como una categoría que a priori no determina los sentidos que las personas usuarias tienen sobre la reproducción y los embriones in vitro.

Current perceptions and decisions of patients about cryopreserved embryos in Argentina. / [Percepciones actuales y decisiones de pacientes acerca de embriones criopreservados en Argentina.]

Introduction: Worldwide there is an increase in the number of assisted reproduction treatments. The developments in culture media and drugs, associated with the occurrence of rapid freezing or vitrification techniques has improved the procedures encouraging transfers of a single embryo per cycle. This generates a greater number of viable surplus embryos. An embryo disposition decision making is a complex phenomenon. Method: Method: This descriptive exploratory study investigates the perceptions and opinions of a sample of 60 participants who have started an assisted reproduction treatment in a specialized Fertility Center of the City of Buenos Aires. Results: The absence of children in the majority of the sample (70.6% still have no children) makes the prevailing option to "continue cryopreservation" (36.67%). The majority, 38.4% (23 people), considers that the option that best represents the embryo is "a child". Although there are relatively few people who have decided to donate their embryos (5 participants), several study participants report having reflected on it (35%). The vast majority (86.6%) of respondents would not be willing to face the costs of maintaining a long-term cryopreservation. Conclusion: After more than three decades of assisted reproduction, Argentinean patients still face a high degree of uncertainty regarding what to do with the surplus embryos. Gaining more in-depth knowledge regarding the decision-making process can help to improve the communication strategies. The normative absences generate that the ARTs users are "disengaged" from their embryos; this hinders the decision making, increasing the risk of accumulating more "abandoned" embryos

Introducción: A nivel mundial se registra un aumento en la cantidad de tratamientos de reproducción asistida. La optimización en medios de cultivo y fármacos, asociado al advenimiento de técnicas de congelamiento rápido o vitrificación han logrado mejorar los procedimientos alentando las transferencias de un único embrión por ciclo, generando un mayor número de embriones excedentes viables. La toma de las decisiones sobre la disposición de los embriones supernumerarios es un fenómeno complejo. Método: Este estudio exploratorio descriptivo indaga las percepciones y opiniones de una muestra de 59 participantes que han comenzado un tratamiento de reproducción asistida en un Centro de Fertilidad especializado de la Ciudad de Buenos Aires. Resultados: La ausencia de hijos en la mayoría de la muestra (70.6% aún no tiene hijos) hace que la opción prevalente sea continuar la criopreservación (35,6%). La mayoría, 40.7% (24 personas) considera que la opción que mejor representa al embrión es "un hijo/a". Si bien son relativamente pocas las personas que han decidido donar sus embriones (5 personas) varios participantes del estudio refieren haber reflexionado sobre ello (43%). La gran mayoría (86,5%) de los encuestados no estaría dispuesta a enfrentar los gastos del mantenimiento de la criopreservación a largo plazo. Conclusión: Se constata que, luego de más de tres décadas de desarrollo de las tecnologías reproductivas, aún existe en Argentina un alto grado de incertidumbre en los pacientes respecto de qué hacer con sus embriones excedentes. Conocer más en profundidad el proceso de toma de decisiones puede contribuir a implementar estrategias comunicacionales.

[Developmental stages and viability of Toxocara canis eggs outside the host].

INTRODUCTION: Toxocariasis is a soil-transmitted zoonotic disease caused mainly by ingestion of larvated eggs of Toxocara canis. OBJECTIVES: To study the morphology of the intraovular developmental stages of Toxocara canis in culture, characterize non-viable eggs and the sequences of larval molting and compare the viability of eggs at the early stages of division and at reaching full maturation. MATERIAL AND METHODS: Observation of developing embryos and characterization of non-viable eggs were done using light microscope. The proportions of viable eggs during embryonation were compared to the proportions of viable mature eggs. RESULTS: Cell division commenced after 24 hours of cultivation. Early stages were found to be present over a period of 3-5 days. The developmental stages identified were eggs with: One cell, two cells, three cells, four cells, early morula, late morula, blastula, gastrula, tadpole, pre-larva, first, second and third stage larva. Two larval molts occurred. Non-viable eggs had degenerated cytoplasm, thin or collapsed shell and the larvae did not move after exposure to light. No significant differences were found between the proportions of viable eggs from day five to day 21 as compared to viability of fully mature eggs (30 days). CONCLUSION: Developing embryos in the environment may be considered as a potential threat to the public health. The precise identification of developmental stages and the clear differentiation of viable and non-viable eggs can help in determining an accurate baseline rate of development that could be used in studies of ovicidal compounds.

Developmental stages and viability of Toxocara canis eggs outside the host / Desarrollo y viabilidad de huevos de Toxocara canis

ABSTRACT Introduction: Toxocariasis is a soil-transmitted zoonotic disease caused mainly by ingestion of larvated eggs of Toxocara canis. Objectives: To study the morphology of the intraovular developmental stages of Toxocara canis in culture, characterize non-viable eggs and the sequences of larval molting and compare the viability of eggs at the early stages of division and at reaching full maturation. Material and methods: Observation of developing embryos and characterization of non-viable eggs were done using light microscope. The proportions of viable eggs during embryonation were compared to the proportions of viable mature eggs. Results: Cell division commenced after 24 hours of cultivation. Early stages were found to be present over a period of 3-5 days. The developmental stages identified were eggs with: One cell, two cells, three cells, four cells, early morula, late morula, blastula, gastrula, tadpole, pre-larva, first, second and third stage larva. Two larval molts occurred. Non-viable eggs had degenerated cytoplasm, thin or collapsed shell and the larvae did not move after exposure to light. No significant differences were found between the proportions of viable eggs from day five to day 21 as compared to viability of fully mature eggs (30 days). Conclusion: Developing embryos in the environment may be considered as a potential threat to the public health. The precise identification of developmental stages and the clear differentiation of viable and non-viable eggs can help in determining an accurate baseline rate of development that could be used in studies of ovicidal compounds.

RESUMEN Introducción. La toxocariasis es una enfermedad zoonótica transmitida por contacto con el suelo contaminado y causada principalmente por la ingestión de huevos larvados de Toxocara canis. Objetivos. Estudiar la morfología de los estadios intraovulares en desarrollo de T. canis en cultivo, caracterizar los huevos no viables y las secuencias de las mudas larvarias, y comparar la viabilidad de los huevos en las etapas tempranas de división y al alcanzar la maduración completa. Materiales y métodos. Se observó el desarrollo de los embriones y se caracterizaron los huevos no viables, mediante microscopía de luz. Se comparó la proporción de huevos viables con embrión con la de huevos maduros viables. Resultados. La división celular comenzó 24 horas después de iniciado el cultivo. Los estadios tempranos estuvieron presentes por un periodo de tres a cinco días. Los estadios de desarrollo identificados fueron: huevos con una célula, con dos células, con tres células y con cuatro células;mórula temprana, mórula tardía, blástula, gástrula, renacuajo, prelarva, primer, segundo y tercer estado larvario. Se presentaron dos mudas larvarias. Los huevos no viables tenían el citoplasma degradado, cubierta exterior delgada o colapsada, y su larva no se movía al exponerla a la luz. No se encontraron diferencias significativas entre la proporción de huevos viables del día 5 al día 21, al compararla con la viabilidad de los huevos completamente maduros (30 días). Conclusión. Los embriones en desarrollo en el medio ambiente pueden considerarse como un riesgo potencial para la salud pública. La identificación precisa de los estadios de desarrollo y la clara diferenciación de huevos viables y no viables, pueden ayudar a determinar con exactitud una tasa basal de desarrollo, la cual sería útil en el estudio de compuestos ovicidas.

Supplementation with sunflower seeds in beef cattle did not impact on oocyte and in vitro embryo production.

Supplementation with compounds rich in linoleic acid, including sunflower seed supplementation, promotes increase in conception rates in cows. We aimed to evaluate whether the sunflower seed (linoleic acid source) supplementation in beef donor females alters the plasma concentrations of cholesterol, triglycerides, HDL and LDL, increases the number and quality of oocytes, increases the cleavage rates and determines an improvement in number and quality of in vitro produced blastocysts. Thus, Nelore females were divided into two groups of 15 animals to receive supplementation with or without sunflower seed for 57 days. Females underwent follicular aspiration and the oocytes were subjected to in vitro embryo production. There was no difference (p > .1) between control group and group supplemented with sunflower seed on the number of displayed follicles; number of aspired oocytes; recovery rate; cleavage rate; number of embryos; number of blastocysts; embryos number of grades I and II; plasma concentrations of total cholesterol, triglycerides; HDL and LDL. Therefore, sunflower seed supplementation in oocyte donors did not increase the number and quality of oocytes, cleavage rates and the number and quality of blastocysts produced in vitro.

Embrioteca: espacio para la investigación científica y la docencia en la universidad médica villaclareña / Embrioteca: space for scientific research and teaching at the medical university of Villa Clara

Las embriotecas humanas son valiosas colecciones de material embrionario humano que honran las instituciones que las poseen. Entre las más citadas actualmente están prestigiosas embriotecas en España, Estados Unidos y Argentina. Resultan de un laborioso trabajo de recolección y procesamiento de delicadas muestras embrionarias con potencialidades de uso docente e investigativo. Han sido múltiples las aportaciones al conocimiento científico a partir del estudio de sus series, además del inestimable valor didáctico de las imágenes que atesoran. La Universidad de Ciencias Médicas de Villa Clara presenta la modesta colección que posee, abierta a nuevas contribuciones y a los interesados en su consulta. El objetivo de esta revisión es exponer su relevancia científica y docente en un contexto de revisión histórica y actualidad.

Human embryos are valuable collections of human embryonic material that honor the institutions that own them. Among the most cited are currently prestigious embryos in Spain, the United States and Argentina. They result from a laborious work of collection and processing of delicate embryonic samples with potential for teaching and research use. There have been many contributions to scientific knowledge from the study of their series, in addition to the invaluable didactic value of the images they treasure. The University of Medical Sciences of Villa Clara presents the modest collection that it has, open to new contributions and to those interested in its consultation. The objective of this review is to expose its scientific and teaching relevance in a context of historical review and current affairs.

[Relationship between mitochondrial DNA copy number, membrane potential of human embryo and embryo morphology].

Objective: To explore the relationship between the embryo with the different morphological types in the third day and its mitochondrial copy number, the membrane potential. Methods: Totally 117 embryos with poor development after normal fertilization and were not suitable transferred in the fresh cycle and 106 frozen embryos that were discarded voluntarily by infertility patients with in vitro fertilization-embryo transfer after successful pregnancy were selected. According to evaluation of international standard in embryos, all cleavage stage embryos were divided into class Ⅰ frozen embryo group (n=64), class Ⅱ frozen embryo group (n=42) and class Ⅲ fresh embryonic group (not transplanted embryos; n=117). Real-time PCR and confocal microscopy methods were used to detect mitochondrial DNA (mtDNA) copy number and the mitochondrial membrane potential of a single embryo. The differences between embryo quality and mtDNA copy number and membrane potential of each group were compared. Results: The copy number of mtDNA and the mitochondrial membrane potential in class Ⅲ fresh embryonic group [(1.7±1.0)×10(5) copy/µl, 1.56±0.32] were significantly lower than those in class Ⅰ frozen embryo group [(3.4±1.7)×10(5) copy/µl, 2.66±0.21] and class Ⅱ frozen embryo group [(2.6±1.2)×10(5) copy/µl, 1.80±0.32; all P<0.05]. The copy number of mtDNA and the mitochondrial membrane potential in classⅠ frozen embryo group were significantly higher than those in classⅡ frozen embryo group (both P<0.05). Conclusion: The mtDNA copy number and the mitochondrial membrane potential of embryos of the better quality embryo are higher.

Relationship between mitochondrial DNA copy number, membrane potential of human embryo and embryo morphology / 中华妇产科杂志

Objective To explore the relationship between the embryo with the different morphological types in the third day and its mitochondrial copy number,the membrane potential.Methods Totally 117 embryos with poor development after normal fertilization and were not suitable transferred in the fresh cycle and 106 frozen embryos that were discarded voluntarily by infertility patients with in vitro fertilization-embryo transfer after successful pregnancy were selected.According to evaluation of international standard in embryos,all cleavage stage embryos were divided into class Ⅰ frozen embryo group (n=64),class Ⅱ frozen embryo group (n=42) and class Ⅲ fresh embryonic group (not transplanted embryos;n=117).Real-time PCR and confocal microscopy methods were used to detect mitochondrial DNA (mtDNA) copy number and the mitochondrial membrane potential of a single embryo.The differences between embryo quality and mtDNA copy number and membrane potential of each group were compared.Results The copy number of mtDNA and the mitochondrial membrane potential in class Ⅲ fresh embryonic group [(1.7± 1.0)× 105 copy/μl,1.56±0.32] were significantly lower than those in class Ⅰ frozen embryo group [(3.4±1.7)×105 copy/μl,2.66±0.21]and class Ⅱ frozen embryo group [(2.6± 1.2)× 105 copy/μl,1.80±0.32;all P＜0.05].The copy number of mtDNA and the mitochondrial membrane potential in class Ⅰ frozen embryo group were significantly higher than those in class Ⅱ frozen embryo group (both P＜0.05).Conclusion The mtDNA copy number and the mitochondrial membrane potential of embryos of the better quality embryo are higher.

Preservação da fertilidade em mulheres com câncer: atualização e perspectivas / Fertility preservation in women with cancer: update and perspectives

Com o aumento do diagnóstico de câncer em mulheres jovens e os avanços no seu tratamento, muitas pacientes que poderão ter sua fertilidade comprometida com a quimioterapia têm manifestado desejo de engravidar futuramente. O congelamento de embriões, após fertilização in vitro, para preservar a fertilidade está bem estabelecido. A criopreservação de oócitos por vitrificação evoluiu bastante nos últimos anos, deixando de ser experimental. Até 2009 nasceram mais de 900 crianças a partir de oócitos criopreservados, sem aumento do risco de anomalias congênitas. O uso de análogos do GnRH para a supressão ovariana durante a quimioterapia na tentativa de prevenir a falência ovariana prematura apresenta resultados incertos. Outras técnicas ainda são consideradas experimentais, como a criopreservação e posterior autotransplante de tecido ovariano. Já foram relatados 24 nascimentos com o seu uso, persistindo, entretanto, dúvidas que motivam o seu estudo. A maturação de folículos ovarianos in vitro é alternativa promissora para preservação da fertilidade nessas pacientes e tem apresentado resultados positivos em roedores, macacos e humanos. Muita cautela deve ser tomada com o uso de técnicas experimentais, especialmente quando oferecidas para pacientes diante de fragilidade emocional. Por isso, é importante transmitir corretamente informações sobre chances de gravidez com tratamentos existentes e as limitações das técnicas experimentais...

With the increased number of cancer diagnoses among young women and the advances in treatment, many patients who may have had their fertility compromised by chemotherapy express desire to become pregnant in the future. Freezing embryos for later IVF so as to preserve fertility is a well established process. Oocyte cryopreservation by vitrification has evolved greatly in recent years and is no longer considered experimental. By 2009 more than 900 children were born from cryopreserved oocytes, without increased risk of congenital anomalies. The preventive use of GnRH analogues for ovarian suppression during chemotherapy to avoid premature ovarian failure has uncertain outcomes. Other techniques such as cryopreservation of ovarian tissue for later autograft are still considered experimental. Although use has already been reported in 24 births, doubts still persist and motivate further study. In vitro maturation of ovarian follicles is a promising alternative for preserving patient fertility and has shown positive results in rodents, monkeys, andhumans. Caution should be used with experimental techniques, especially when offered to emotionally fragile patients. Therefore it is important to thoroughly convey information on the chances of pregnancy with existing treatments and the limitations of experimental techniques...

E16.5 embryonic mouse pancreatic tissue transplantation for the treatment of experimental diabetes / 中华普通外科杂志

Objective To investigate the effect of early embryonic mouse pancreatic tissue transplantation in the treatment of mouse experimental diabetes.Methods C57BL/6 mouse models of streptozocin-induced diabetes mellitus were established and then randomly divided into two groups:transplantation group,in which,five to seven pieces of pancreatic tissue of mice at embryonic 16.5 days were transplanted into mouse renal capsule,and sham-operated (SO) control group,in which,0.05 ml RPMI1640 culture medium was injected into mouse renal capsule.When blood glucose level of the transplantation group mouse was ≤ 11.2 mmol/L,the endocrine function of embryonic pancreatic tissue transplanted was detected by IPGTT and IPITT methods and then the transplanted graft was removed for observing the blood glucose surge.In vitro,levels of insulin secretion were measured in serum,E16.5 pancreatic tissue and graft (3.3 mmol/L and 16.7 mmol/L) glucose stimulation by ELISA.Histology and immunohistochemistry were observed before or after pancreatic tissue transplantation to detect insulin and glycagon production.Results (1) The glucose level decreased significantly in the TX group after 4-6 weeks of transplantation (13.4 ± 6.5 vs 28.9 ± 2.5,P ＜ 0.05),and the body weight gaining (P ＜ 0.05).Compared to SO group,glucose level decreased significantly (P ＜ 0.001).The graft can modulate insulin secretion; (2) Insulin can be detected in E16.5 embryonic mouse.Insulin and glycagon were produced more in TX group than pretransplantated pancreatic tissue.Conclusions E16.5 embryonic mouse pancreatic tissue transplantation effectively controls serum glucose level and restore the nondiabetic pattern of weight gaining in diabetic mouse.

Prenatal ultrasound of fetus with echogenic bowel and complicated structural abnormalities/chromosome abnormality / 中华医学超声杂志（电子版）

Objective To explore the prenatal ultrasonographic features of fetuses diagnosed as echogenic bowel and chromosomal abnormalities. Methods From September 2009 to June 2013, eighty cases diagnosed as echogenic bowel by prenatal ultrasound screening in our hospitals underwent chromosomal karyotype analysis and were followed up till fetal birth. The prenatal ultrasonographic features were carefully correlated with the postnatal findings. Results Prenatal ultrasoundand chromosomal findings of the 80 cases were:(1) Fifty-eight cases of fetuses with echogenic bowel alone (72.5%, 58/80, without chromosomal karyotype abnormalities). (2) Eleven cases of fetuses with additional ultrasound soft markers (13.8%, 11/80, without chromosomal karyotype abnormalities). The ultrasound examination revealed 8 cases of choroid plexus cysts, 1 case of single umbilical artery, 1 case of shorter nasal bone, and 1 case of echogenic intracardiac focus. (3) Five cases of fetuses with severe functional abnormalities or complex structural malformations and normal chromosomal karyotype. The ultrasound examination revealed 3 cases of structural cardiac malformations, 1 case of heart failure and 2 cases of central nervous system malformations. Of them,one case showed complex malformations (right ventricular dysplasia syndrome, cerebellar dysplasia and single umbilical artery), whereas the remaining 4 cases only involve single malformation. (4)Six cases had chromosomal abnormalities (7.5%, 6/80), including 3 cases of abnormal chromosome microdeletions, and 3 cases of abnormal chromosome numbers (1 21-trisomy syndrome, 1 18-trisomy syndrome, and 1 triploid syndrome). All of them had severe structural malformations and abnormal soft markers. The ultrasound examination revealed 2 cases of severe cardiac malformations, 2 cases of central nervous system malformations, 1 case of pericardial/peritoneal effusion, and 1 case of placental chorionic multiple hemangioma. One case of holoprosencephaly (donor of the twins) ended with intrauterine death. The associated abnormal soft markers include nuchal fold (NF) thickening, choroid plexus cysts, umbilical cord cysts, single umbilical artery, and placental chorionic hemangioma. Conclusions The fetuses with echogenic bowelaloneusualyhaveagoodprognosis.Insomecases,echogenicbowelmaydisappearduringvfolow-up.The fetuses complicated with severe structural malformations and chromosomal abnormalities have poor prognosis. For the fetuses diagnosed of echogenic bowel by prenatal ultrasound screening, systematic ultrasound examination should be carried out and clinical chromosomal karyotype analysis was suggested.

Odontogenic cell culture in PEGDA hydrogel scaffolds for use in tooth regeneration protocols

In order to obtain a tooth-like structure, embryonic oral ectoderm cells (EOE) and bone marrow-derived stem cells (BMSC) were stratified within a synthetic hydrogel matrix (PEGDA) and implanted in the ileal mesentery of adult male Lewis rats. Wholemount in situ hybridization was used to evaluate the expression of Pitx2, Shh and Wnt10a signals indicative of tooth initiation. In rats, expression of the three markers was present in the oral ectoderm starting at embryonic stage E12.5, which was therefore selected for cell harvesting. Embryos were obtained by controlled service of young female Lewis rats in which estrus was detected by impedance reading. At E12.5, pregnant rats were humanely euthanized and embryos were collected. The mandibular segment of the first branchial arch was dissected and the mesenchyme separated from the ectoderm by enzymatic digestion with pancreatin trypsin solution. BMSCs were collected by flushing the marrow of tibiae and femurs of adult Lewis rats with a-MEM and cultured in a-MEM in 25 cm2 flasks. Second passage BMSC's were recombined with competent oral ectoderm (E12.5-E13) stratifying them within a 3D PEGDA scaffold polymerized by exposure to UV (365nm) inside a pyramidal polypropylene mold. Constructs were incubated from 24 to 48 hrs in a-MEM and then implanted for four to six weeks in the mesentery of adult male (3- 6 month old) Lewis rats. 76 constructs were implanted (37 experimental, 27 negative controls and 12 positive controls). Upon maturation, constructs were harvested, fixed in buffered formalin, processed and stained with hematoxylin eosin (HE). Histological evaluation of the experimental and negative constructs showed that BMSCs underwent an apoptotic process due to lack of matrix interactions, known as anoikis, and were thus incapable of interacting with the competent ectoderm. In contrast, embryonic oral ectoderm was able to proliferate during the mesenteric implantation. In conclusion, PEGDA scaffolds are incompatible with BMSCs, therefore it is essential to continue the search for an ideal scaffold that allows proper tissue interactions.

Para lograr la formacion de una estructura similar a un diente se estratificaron celulas de ectodermo oral embrionario (EOE) con celulas troncales de medula osea (BMSC) dentro de una matriz de diacrilato de polietilenglicol (PEGDA) que se implanto en el mesenterio ileal de machos adultos de ratas Lewis. Mediante hibridizacion in situ de bloque completo se evaluo la expresion de tres genes iniciadores putativos de la formacion dental (Pitx2, Shh y Wnt10a), estableciendo que en ratas las senales iniciadoras de dentogenesis aparecen entre E12.5 y E13. El tejido ectodermico embrionario se obtuvo haciendo cruces controlados de hembras a las que se les detecto el estro mediante impedanciometria. En E12.5 las hembras se sacrificaron y se extrajeron los embriones. Se diseco la porcion mandibular del primer arco branquial y el ectodermo se separo del mesenquima mediante disociacion enzimatica con una solucion de pancreatina tripsina. Las BMSC se extrajeron de los huesos largos de las extremidades inferiores de ratas mediante lavado con a-MEM y se cultivaron en cajas de 25cm2 hasta un segundo pasaje. Las BMSC fueron recombinadas con ectodermo embrionario competente (E12.5 - E13) estratificandolas en un soporte tridimensional de PEGDA, polimerizado con luz ultravioleta (365nm) dentro de un molde piramidal de polipropileno (PP). Los constructos se cultivaron entre 48 y 72 horas en a-MEM y posteriormente fueron implantados en el mesenterio ileal de machos adultos (3 a 6 meses de edad) por un periodo de cuatro a seis semanas. Se implantaron 76 constructos (37 experimentales, 27 controles negativos y 12 controles positivos). En la fecha determinada los animales se sacrificaron mediante asfixia con una mezcla de CO2 y aire recuperando los constructos que se fijaron en formalina tamponada para luego procesarlos y tenirlos con hematoxilina eosina (HE). La evaluacion histologica de los constructos experimentales, positivos y negativos mostro que las BMSC incluidas en el hidrogel sufrieron un proceso de apoptosis conocido como anoikis que impidio su interaccion con las celulas ectodermicas. En contraste el EOE prolifero durante el periodo de implantacion. A futuro se debe buscar la matriz portadora ideal que permita el confinamiento de los dos grupos celulares y que brinde el soporte estructural necesario para la proliferacion de las BMSC facilitando su interaccion con las celulas inductoras de origen ectodermico.