Características clínicas en pacientes con menos de 12 folículos ováricos como hallazgo ecográfico / Clinical and ultrasound findings in patients with ovaries with a count of less than 12 follicles

Introducción y objetivos: Determinar los hallazgos clínicos y ecográficos en pacientes que presentan menos de 12 folículos ováricos. Método: Estudio observacional (cohorte histórica) con 505 pacientes seleccionadas mediante muestreo consecutivo, entre el 14 de enero del 2019 y el 15 de marzo del 2021, que consultan por diversas alteraciones ginecológicas. Se generan dos grupos de pacientes, las que presentaron uno a tres folículos en uno de los ovarios (n = 377) y las que presentaban 4 a 11 folículos (n = 128). Se midió como resultado primario la presencia de al menos un signo clínico de hiperandrogenismo. Resultados: De 505 pacientes analizadas, al comparar las que presentaron 4 a 11 folículos en uno de los ovarios (n = 377) con las que presentaban 1 a 3 folículos (n = 128), las primeras mostraron mayor presencia de signos de hiperandrogenismo, endometrio en fase lútea de mayor espesor y un patrón menstrual con uno a cuatro días de sangrado menstrual abundante, diferencias todas estadísticamente significativas (p < 0,05). Conclusión: En pacientes con 4 a 11 folículos en uno de sus ovarios, se observaron signos de hiperandrogenismo, similares al síndrome de ovario poliquístico.

Introduction and objectives: Determine the clinical and ultrasound findings in patients who present less than 12 ovarian follicles in the ultrasound count. Method: Observational study (historical cohort) with 505 patients selected by consecutive sampling, between January 14, 2019 and March 15, 2021, who consulted for different gynecological disorders. Two groups of patients were generated: those with 1 to 3 follicles in one of the ovaries (n = 377) and those with 4 to 11 follicles (n = 128). The primary outcome was the presence of at least one clinical sign of hyperandrogenism. Results: Of 505 patients analyzed, when comparing those who presented 4 to 11 follicles in one of the ovaries (n = 377) with those who presented 1 to 3 follicles (n = 128), the first group showed a greater presence of signs of hyperandrogenism, thicker endometrium in luteal phase and a menstrual pattern with one to four days of heavy menstrual bleeding, all differences were statistically significant (p < 0.05). Conclusion: In patients with 4 to 11 follicles in one of their ovaries, signs of hyperandrogenism, similar to polycystic ovary syndrome, were observed.

Manejo del Síndrome de Hiperestimulación Ovárica / Management of Ovarian Hyperstimulation Syndrome

INTRODUCCIÓN: El síndrome de hiperestimulación ovárica es una respuesta exagerada del ovario a los tratamientos hormonales para estimular la formación de óvulos. OBJETIVO: Describir el caso clínico de una mujer con síndrome de hiperestimulación ovárica; revisar el abordaje, manejo, tratamiento y cómo prevenirlo. CASO CLÍNICO: Paciente femenina de 37 años, multigesta, en tratamiento con metformina por Síndrome de ovario poliquístico , que presenta infertilidad secundaria a factor tubárico, que desarrolló un cuadro moderado de síndrome de hiperestimulación ovárica como consecuencia de la aplicación de las técnicas de fertilización in vitro (Folitropina alfa humana recombinante (GONAL-F®) y Cetrolerelix (CETROTIDE®); al cuarto día del procedimiento de aspiración folicular presenta dolor pélvico intenso, disuria, deposiciones diarreicas, ecografía abdominal y vaginal evidencia líquido libre en cavidad alrededor de 1000cc, además de ovarios tanto derecho e izquierdo con volumen de 102 mL y 189 mL respectivamente. Paciente es ingresada para realizar tratamiento hidratación parenteral, Enoxaparina 40mg subcutánea, Cabergolina 0.5mg vía oral, alta a las 72 horas. DISCUSIÓN: Las claves para la prevención del síndrome de hiperestimulación ovárica son la experiencia con la terapia de inducción de la ovulación y el reconocimiento de los factores de riesgo para el síndrome de hiperestimulación ovárica. Los regímenes de inducción de la ovulación deberían ser altamente individualizados, monitorizados cuidadosamente y usando dosis y duración mínimas del tratamiento con gonadotropinas para conseguir la meta terapéutica. CONCLUSIONES: El síndrome de hiperestimulación ovárica constituye la complicación más temida durante el uso de inductores de la ovulación; el conocimiento de factores de riesgo, puede prevenir o evitar que llegue a ser de un caso severo, lo cual puede causar mayor morbilidad o hasta mortalidad. La vitrificación se convierte en la técnica que permite prevenir el síndrome de hiperestimulación ovárica, junto con esta técnica hay 2 alternativas: la inducción con análogo de la hormona liberadora de gonadotropina o el uso de agonistas dopaminérgicos.

INTRODUCTION: Ovarian hyperstimulation syndrome is an exaggerated response of the ovary to hormonal treatments to stimulate egg formation. OBJECTIVE: To describe the clinical case of a woman with ovarian hyperstimulation syndrome; to review the approach, management, treatment and how to prevent it. CLINICAL CASE: 37-year-old female patient, multigestation, under treatment with metformin for polycystic ovary syndrome, presenting infertility secondary to tubal factor, who developed a moderate picture of ovarian hyperstimulation syndrome as a consequence of the application of in vitro fertilization techniques (recombinant human follitropin alfa (GONAL-F®) and Cetrolerelix (CETROTIDE®); On the fourth day of the follicular aspiration procedure she presents intense pelvic pain, dysuria, diarrheic stools, abdominal and vaginal ultrasound shows free fluid in the cavity of about 1000cc, in addition to right and left ovaries with a volume of 102 mL and 189 mL respectively. Patient was admitted for parenteral hydration treatment, Enoxaparin 40mg subcutaneous, Cabergoline 0.5mg orally, discharged after 72 hours. DISCUSSION: The keys to prevention of ovarian hyperstimulation syndrome are experience with ovulation induction therapy and recognition of risk factors for ovarian hyperstimulation syndrome. Ovulation induction regimens should be highly individualized, carefully monitored, and using minimal doses and duration of gonadotropin therapy to achieve the therapeutic goal. CONCLUSIONS: Ovarian hyperstimulation syndrome constitutes the most feared complication during the use of ovulation inducers; knowledge of risk factors, may prevent or avoid it from becoming a severe case, which may cause increased morbidity or even mortality. Vitrification becomes the technique that allows preventing ovarian hyperstimulation syndrome, along with this technique there are 2 alternatives: induction with gonadotropin-releasing hormone analog or the use of dopaminergic agonists.

Distocia em tigre d'água-de-orelha-vermelha, Trachemys scripta elegans, wied (1839): relato de caso / Dystocia in red-eared slider, Trachemys scripta elegans, wied (1839): case report / Distocia en tortuga de orejas rojas, Trachemys scripta elegans, wied (1839): reporte de caso

Atualmente muitos répteis se tornaram animais de companhia e são mantidos como pet's exóticos. A espécie Trachemys scripta elegans, Wied (1839) é um animal exótico da América do Norte, sua identificação é realizada pelas marcas avermelhadas encontradas lateralmente a sua cabeça. Na rotina clínica as principais enfermidades que acometem os quelônios são as de origem reprodutiva, como a estase folicular e distocia. O objetivo deste trabalho foi relatar um caso recorrente de distocia em um tigre d'água fêmea, para isso, a anamnese, o histórico da paciente, e seus sinais clínicos, em conjunto com os exames complementares de imagem foram essenciais para se obter diagnóstico definitivo. O tratamento foi realizado com a indução medicamentosa utilizando borogluconato de cálcio, seguida da aplicação de ocitocina, esta trouxe resultados positivos para a eliminação dos ovos. Porém devido ao histórico do paciente, optou-se pela intervenção cirúrgica de ovariossalpingectomia, sendo está a maneira permanente de resolução da patologia. O protocolo terapêutico escolhido proporcionou um resultado satisfatório e bem estar ao animal.(AU)

Currently, many reptiles have become companion animals and are kept as exotic pets. The species Trachemys scripta elegans, Wied (1839) is an exotic animal from North America, and its identification is based on the reddish markings found laterally on its head. In routine clinical practice, the main diseases that affect chelonians are those of reproductive origin, such as follicular stasis and dystocia. The aim of this study was to report a recurrent case of dystocia in a female red-eared slider turtle. For this purpose, the patient's anamnesis, history, and clinical signs, along with complementary imaging exams, were essential to obtain a definitive diagnosis. The treatment involved medical induction using calcium borogluconate, followed by the administration of oxytocin, which yielded positive results in egg elimination. However, due to the patient's history, surgical intervention in the form of ovariosalpingectomy was chosen as the permanent solution to the pathology. The chosen therapeutic protocol provided a satisfactory outcome and improved the animal's well-being.(AU)

Actualmente muchos reptiles se han convertido en animales de compañía y se mantienen como mascotas exóticas. La especie Trachemys scripta elegans, Wied (1839) es un animal exótico de América del Norte, su identificación se realiza por las marcas rojizas que se encuentran lateralmente a su cabeza. En la rutina clínica, las principales enfermedades que afectan a los quelonios son las de origen reproductivo, como la estasis folicular y la distocia. El objetivo de este trabajo fue reportar un caso recurrente de distocia en una hembra de tigre de agua, para ello la anamnesis, la historia de la paciente y sus signos clínicos, junto con los exámenes imagenológicos complementarios fueron fundamentales para obtener un diagnóstico definitivo. El tratamiento se realizó con inducción farmacológica con borogluconato de calcio, seguido de la aplicación de oxitocina, que arrojó resultados positivos con la eliminación de huevos. Sin embargo, debido a los antecedentes de la paciente, se optó por la intervención quirúrgica de ovarialpingectomía, que es la forma definitiva de resolución de la patología. El protocolo terapéutico elegido proporcionó un resultado satisfactorio y bienestar al animal.(AU)

Effect of Liangfang Wenjing Decoction on expression of key glycolytic enzymes in uterus and ovaries of rats with coagulating cold and blood stasis syndrome / 中国中药杂志

This study aimed to investigate the relationship between coagulating cold and blood stasis syndrome and glycolysis, and observe the intervention effect of Liangfang Wenjing Decoction(LFWJD) on the expression of key glycolytic enzymes in the uterus and ovaries of rats with coagulating cold and blood stasis. The rat model of coagulating cold and blood stasis syndrome was established by ice-water bath. After modeling, the quantitative scoring of symptoms were performed, and according to the scoring results, the rats were randomly divided into a model group and LFWJD low-, medium-and high-dose groups(4.7, 9.4, 18.8 g·kg~(-1)·d~(-1)), with 10 in each group. Another 10 rats were selected as the blank group. After 4 weeks of continuous administration by gavage, the quantitative scoring of symptoms was repeated. Laser speckle flowgraphy was used to detect the changes of microcirculation in the ears and uterus of rats in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological morphology of uterus and ovaries of rats in each group. The mRNA and protein expressions of pyruvate dehydrogenase kinase 1(PDK1), hexokinase 2(HK2) and lactate dehydrogenase A(LDHA) in the uterus and ovaries of rats were examined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot, respectively. The rats in the model group showed signs of coagulating cold and blood stasis syndrome, such as curl-up, less movement, thickened veins under the tongue, and reduced blood perfusion in the microcirculation of the ears and uterus, and HE staining revealed a thinning of the endometrium with disorganized arrangement of epithelial cells and a decrease in the number of ovarian follicles. Compared with the model group, the treatment groups had alleviated coagulating cold and blood stasis, which was manifested as red tongue, reduced nail swelling, no blood stasis at the tail end as well as increased blood perfusion of the microcirculation in the ears and uterus(P<0.05 or P<0.01). Among the groups, the LFWJD medium-and high-dose groups had the most significant improvement in coagulating cold and blood stasis, with neatly arranged columnar epithelial cells in uterus, and the number of ovarian follicles was higher than that in the model group, especially mature follicles. The mRNA and protein expressions of PDK1, HK2, LDHA in uterus and ovaries were up-regulated in the model group(P<0.05 or P<0.01), while down-regulated in LFWJD medium-and high-dose groups(P<0.05 or P<0.01). The LFWJD low-dose group presented a decrease in the mRNA expressions of PDK1, HK2 and LDHA in uterus and ovaries as well as in the protein expressions of HK2 and LDHA in uterus and HK2 and PDK1 in ovaries(P<0.05 or P<0.01). The therapeutic mechanism of LFWJD against coagulating cold and blood stasis syndrome is related to the down-regulation of key glycolytic enzymes PDK1, HK2 and LDHA, and the inhibition of glycolytic activities in uterus and ovaries.

Research progress on the biological effects of HIF-1α on follicle development and ovulation / 生理学报

Hypoxia inducible factor-1α (HIF-1α), as a hypoxia inducible factor, affects women's reproductive function by regulating the development and excretion of follicles. HIF-1α induces glycolysis and autophagy in the granule cells by promoting oocyte development, regulating the secretion of related angiogenic factors, and improving follicle maturity. In addition, HIF-1α promotes the process of luteinization of follicular vesicles, maintains luteal function, and finally completes physiological luteal atrophy through cumulative oxidative stress. Dysfunction of HIF-1α will cause a series of pathological consequences, such as angiogenesis defect, energy metabolism abnormality, excessive oxidative stress and dysregulated autophagy and apoptosis, resulting in ovulation problem and infertility. This article summarizes the previous studies on the regulation of follicle development and excretion and maintenance of luteal function and structural atrophy by HIF-1α. We also describe the effective intervention mechanism of related drugs or bioactive ingredients on follicular dysplasia and ovulation disorders through HIF-1α, in order to provide a systematic and in-depth insights for solving ovulation disorder infertility.

Research Progress in the Regulation of Follicle Development by Melatonin / 中国医学科学院学报

Melatonin,an endocrine hormone synthesized by the pineal gland,plays an important role in the reproduction.The growth and development of follicles is the basis of female mammalian fertility.Follicles have a high concentration of melatonin.Melatonin receptors exist on ovarian granulosa cells,follicle cells,and oocytes.It regulates the growth and development of these cells and the maturation and atresia of follicles,affecting female fertility.This paper reviews the protective effects and regulatory mechanisms of melatonin on the development of ovarian follicles,granulosa cells,and oocytes and makes an outlook on the therapeutic potential of melatonin for ovarian injury,underpinning the clinical application of melatonin in the future.

Preservação de fertilidade: cultivo de folículos ovarianos e novo dispositivo para criopreservação seminal / Fertility preservation: ovarian follicle culture and a new device for seminal cryopreservation

INTRODUÇÃO: A oncofertilidade tem o desafio de buscar estratégias para preservar a função reprodutiva. Este estudo explorou duas possibilidades como implementação para as técnicas de preservação da fertilidade feminina e masculina. OBJETIVO: Analisar a eficiência do cultivo de folículos pré-antrais de camundongos suplementado com lisado de plaquetas humanas e desenvolver um protótipo para criopreservação de sêmen humano. MATERIAIS E MÉTODOS: Os folículos pré-antrais foram isolados mecanicamente de ovários de fêmeas de camundongos e foram cultivados individualmente em sistema entre camadas de óleo mineral. Os folículos foram cultivados divididos em 4 grupos, sendo um controle, sem o uso do lisado de plaquetas e três grupos com diferentes concentrações de lisado de plaquetas humanas (PLTMax®). Foram avaliadas a sobrevivência celular, desenvolvimento folicular e características oocitárias. Para o segundo estudo foi desenvolvido e impresso em 3D com filamentos de acrilonitrilo butadieno-estireno (ABS) um protótipo que suporte 10 palhetas com amostras seminais no vapor de nitrogênio líquido (N2L), etapa essencial para criopreservação de sêmen humano. Para os testes foram utilizadas 40 amostras seminais. A temperatura ambiente e no interior das palhetas de envase das amostras foram medidas e estabelecida a curva de resfriamento. Os parâmetros de motilidade, vitalidade e fragmentação do DNA espermático foram avaliados antes do congelamento e após o descongelamento. Foram realizados dois testes, um de posicionamento das palhetas e outro comparativo entre o protótipo e um dispositivo com suporte em poliestireno expandido (EPS). RESULTADOS: O cultivo de 11 dias induziu um aumento no tamanho folicular em todas as condições, sendo maior no grupo controle, seguido do grupo com 10% de PLTMax®, mas com diferença significativa (p<0,001). O grupo controle apresentou maior número de oócitos intactos (>50%) em relação aos demais (<35%). Todos os 4 grupos apresentaram taxas de vitalidade celular acima de 70%. Quanto aos testes com o protótipo em ABS foi verificado que as curvas de refrigeração foram notavelmente reproduzíveis. O material do protótipo resistiu a inúmeros mergulhos (>300) no N2L, sem demonstrar danos ao material. Diferenças significativas (p<0,001) foram observadas para a taxa de recuperação média da motilidade e vitalidade espermática em relação aos dados da amostra 2 fresca em ambos os testes. A motilidade, a vitalidade e a fragmentação do DNA espermático antes do congelamento e após o descongelamento não mostraram diferenças em relação a posição das palhetas. Também não houve diferença quanto ao índice de fragmentação verificada das amostras criopreservadas com uso do protótipo em ABS e o suporte em EPS, mesmo após o cultivo, após 24 horas de cultivo. Contudo, houve diferença em relação a amostra fresca (p<00,1). Quanto a recuperação das taxas de motilidade e vitalidade não houve diferença entre o ABS e EPS após o descongelamento e 24 horas de cultivo. CONCLUSÃO: O PLTMax®, embora tenha apresentado menor desempenho que o HSA, é um candidato de suplementação para o cultivo de folículos pré-antrais que merece ser mais explorado. O protótipo em ABS demonstrou resistência, praticidade e segurança para criopreservação seminal de forma reprodutível e eficiente.

INTRODUCTION: Oncofertility has the challenge of seeking strategies to preserve reproductive function. This study explored two possibilities as implementations for female and male fertility preservation techniques. PURPOSE: To analyze the efficiency of mouse preantral follicle culture supplemented with human platelet lysate and to develop a prototype for human semen cryopreservation. MATERIAL AND METHODS: Preantral follicles were mechanically isolated from female mouse ovaries and were individually cultured using a mineral oil interlayer system. The follicles were cultured divided into 4 groups, one control, without the use of platelet lysate and three groups with different concentrations of human platelet lysate (PLTMax®). Cell survival, follicular development and oocyte characteristics were evaluated. For the second study, a prototype was developed and printed in 3D with acrylonitrile butadiene styrene (ABS) filaments to support 10 straws with seminal samples in liquid nitrogen (N2L) vapor, an essential step for human semen cryopreservation. For the tests 40 seminal samples were used. Ambient and internal temperatures inside the sample straws were measured and the cooling curve was established. The parameters of motility, vitality and sperm DNA fragmentation were evaluated before freezing and after thawing. Two tests were performed, one for positioning the straws and the other comparing the prototype and a device with expanded polystyrene (EPS) support. RESULTS: The 11-day culture induced an increase in follicular size in all conditions, being higher in the control group followed by the group with 10% PLTMax®, but with significant difference (p<0.001). The control group presented a higher number of intact oocytes (>50%) compared to the others (<35%). All 4 groups presented cell vitality rates above 70%. As for the ABS prototype tests, it was verified that the cooling curves were remarkably reproducible. The prototype withstood numerous dips (>300) in N2L without showing damage to the material. Significant differences (p<0.001) were observed for the mean recovery rate of sperm motility and vitality compared to the fresh sample data in both tests. Motility, vitality and sperm DNA fragmentation before freezing and after thawing showed no differences with respect to the position of the straws. There was also no difference in the fragmentation index verified for samples cryopreserved using the ABS prototype and the EPS support, even after 24 hours of culture. However, there was a difference compared to the fresh 4 sample (p<00.1). As for the recovery of motility and vitality rates there was no difference between ABS and EPS after thawing and 24 hours of culture. CONCLUSION: PLTMax®, although it showed lower performance than HSA, is a supplementation candidate for preantral follicle culture that deserves further exploration. The ABS prototype demonstrated strength, practicality and safety for seminal cryopreservation in a reproducible and efficient manner.

Manipulação de oócitos inclusos em folículos ovarianos pré-antrais e a importância dos antioxidantes ­ revisão de literatura / Manipulation of oocytes included in preantral ovarian follicles and the importance of antioxidants ­ literature review / Manipulación de ovocitos incluidos en folículos ováricos preantrales e importancia de los antioxidantes - revisión de la literatura

A manipulação de oócitos inclusos em folículos ovarianos pré-antrais (MOIFOPA) vem sendo estudada pensando na perspectiva futura de aplicação direta na reprodução humana, principalmente para mulheres que sofrem de doenças ou que precisam passar por tratamentos que interferem na função ovariana. Nesse contexto, o objetivo deste trabalho é revisar aspectos relacionados com a biotécnica de MOIFOPA e a importância dos antioxidantes no cultivo in vitro de folículos pré-antrais. Foi realizada uma pesquisa na base de dados PubMed, buscando artigos sobre a biotécnica, principalmente relacionados com a necessidade do uso de antioxidantes no cultivo. A grande maioria dos estudos sobre a biotécnica utilizam como modelo experimental os folículos ovarianos de diferentes espécies de animais. A MOIFOPA compreende o isolamento e o resgate de folículos ovarianos pré-antrais provenientes de ovários, seguido da conservação através da técnica de resfriamento ou congelação e o cultivo folicular in vitro, a fim de promover o crescimento, a maturação e a fecundação in vitro (FIV) dos oócitos inclusos nesses folículos, maximizando o potencial reprodutivo feminino e diminuindo a atresia folicular que acontece in vivo. Um aspecto que pode interferir no sucesso do cultivo in vitro de folículos ovarianos pré-antrais é a produção em excesso de espécies reativas de oxigênio (EROs). Os ácidos ascórbico e alfa lipóico vem demonstrando resultados interessantes para reduzir os efeitos que as EROs causam sobre os folículos ovarianos pré-antrais cultivados in vitro.

The manipulation of oocytes included in preantral ovarian follicles (MOEPF) has been studied considering the future perspective of direct application in human reproduction, especially for women who suffer from diseases or who need to undergo treatments that interfere with ovarian function. In this context, the objective of this paper is to review aspects related to the biotechnology of MOIFOPA and the importance of antioxidants. A search was carried out in the PubMed database, searching for articles on biotechnology, mainly related to the need to use antioxidants in cultivation. The vast majority of studies on biotechnology use ovarian follicles from different species of animals as an experimental model. MOIFOPA comprises the isolation and rescue of preantral ovarian follicles from ovaries, followed by conservation through the cooling or freezing technique and in vitro follicular cultivation, in order to promote growth, maturation and in vitro fertilization ( IVF) of the oocytes included in these follicles, maximizing the female reproductive potential and decreasing the follicular atresia that occurs in vivo. One aspect that may interfere with the success of in vitro culture of preantral ovarian follicles is the excess production of reactive oxygen species (ROS). Ascorbic and alpha lipoic acids have shown interesting results in reducing the effects that ROS cause on in vitro cultured preantral ovarian follicles.

manipulación de ovocitos incluidos en folículos ováricos preantrales (MOIFOPA) se ha estudiado con la perspectiva futura de su aplicación directa en la reproducción humana, especialmente en mujeres que padecen enfermedades o que necesitan someterse a tratamientos que interfieren en la función ovárica. En este contexto, el objetivo de este trabajo es revisar los aspectos relacionados con la biotécnica de MOIFOPA y la importancia de los antioxidantes en el cultivo in vitro de los folículos pré-antrais. Se realizó una investigación en la base de datos PubMed, buscando artículos sobre la biotecnología, principalmente relacionados con la necesidad del uso de antioxidantes en el cultivo. La mayoría de los estudios sobre biotecnología utilizan como modelo experimental los folículos ováricos de diferentes especies de animales. El MOIFOPA incluye el aislamiento y rescate de los folículos ováricos preantrales de los ovarios, seguido de su conservación mediante la técnica de enfriamiento o congelación y el cultivo folicular in vitro, con el fin de promover el crecimiento, la maduración y la fecundación in vitro (FIV) de los ovocitos incluidos en estos folículos, maximizando el potencial reproductivo femenino y disminuyendo la atresia folicular que se produce in vivo. Un aspecto que puede interferir en el éxito del cultivo in vitro de folículos ováricos preantrales es la producción excesiva de especies reactivas de oxígeno (ROS). El ácido ascórbico y el ácido alfa lipoico han mostrado resultados interesantes para reducir los efectos que causan las ERO en los folículos ováricos preantrales cultivados in vitro.

Low versus high antral follicle count on the fertility of timed AI Nelore heifers / Baixa versus alta contagem de folículos antrais na fertilidade de novilhas Nelore inseminadas em tempo-fixo

This study aimed to determine the effect of antral follicle count (AFC), and pubertal status on the fertility of beef heifers. In this study, 230 Nelore heifers, 20±2 months of age, were subjected to an estradiol progesterone-based timed artificial insemination (TAI) program. On Day 0 of the TAI protocol, the heifers were examined by transrectal ultrasound to record videos of the ovaries. Later, in the darkroom of the laboratory of images, the videos were analyzed for AFC (≥ 3 mm) of each ovary. Females who failed the first TAI were resynchronized with the same hormonal protocol. The pregnancy status was evaluated by ultrasonography 30 days after each FTAI. The general mean of the AFC was 22.0 follicles. Thus, the heifers were divided into 2 groups according to AFC: Low AFC (˂ 22 follicles, n = 114), and High AFC (≥ 22 follicles, n = 116). No differences (P > 0.05) in the pregnancy per AI (P/AI) were observed between the Low and High AFC groups, and between pubertal and prepubertal categories. The P/AI was not different between heifers that displayed or did not estrus (P = 0.2). However, considering the estrus response of each AFC group, High AFC heifers that displayed estrus had greater P/AI (P = 0.01) than High AFC heifers that did not display estrus. In summary, AFC and pubertal status did not affect the fertility of Nelore heifers. In contrast, the P/AI of heifers that did not display estrus was lower than heifers observed in estrus only in the High AFC group.(AU)

Esse estudo teve como objetivo determinar o efeito da contagem de folículos antrais (CFA) e da maturidade sexual na fertilidade de novilhas de corte. Neste estudo, 230 novilhas Nelore, com 20 ± 2 meses de idade, foram submetidas a um protocolo de inseminação em tempo-fixo (IATF) a base de estradiol e progesterona. No Dia 0 do protocolo de IATF as novilhas foram examinadas por ultrassonografia transretal e vídeos dos ovários foram gravados para posterior CFA (≥ 3 mm) realizada na sala escura do laboratório de imagens. Trinta dias após a ultrassonografia, as fêmeas que falharam na primeira IATF foram ressincronizadas com o mesmo protocolo hormonal. A prenhez foi avaliada por ultrassonografia 30 dias após cada IATF. A média geral da CFA foi de 22 folículos; assim, as novilhas foram divididas em 2 grupos de acordo com a CFA: CFA baixa (˂ 22 folículos, n=114) e CFA alta (≥ 22 folículos, n=116). A prenhez por IA (P/IA) foi semelhante (P > 0,05) entre os grupos CFA baixa e alta e entre novilhas púberes e pré-púberes. A P/IA não foi diferente entre as novilhas que apresentaram ou não cio (P = 0,2). No entanto, novilhas com CFA alta que apresentaram cio tiveram maior P/IA (P = 0,01) do que novilhas com CFA alta que não apresentaram cio. Em conclusão, a CFA e a maturidade sexual não afetaram a fertilidade de novilhas. Por outro lado, a P/IA das novilhas que apresentaram cio foi maior do que das novilhas não observadas em cio apenas no grupo CFA alta.(AU)

El consumo semicrónico moderado de alcohol altera la foliculogénesis y la calidad núcleo-citoplasmática oocitaria, en el ratón / Moderate semi-chronic alcohol consumption alters folliculogenesis and oocyte nuclear-cytoplasmic quality in mice

Resumen El consumo crónico de alcohol es un problema de salud mundial que afecta particularmente a la población femenina. Sin embargo, los efectos de la ingesta semicrónica en cantidades moderadas a bajas en el ovario y el oocito son poco conocidos. En un modelo murino, se administró etanol al 10% en agua de bebida (hembras tratadas) o agua (hembras control) por 15 días, y luego de la superovulación o no (ovulación espontánea), se analizó el ciclo estral y la calidad ovárico-gamética. En las hembras tratadas, la frecuencia y duración del diestro aumentó, y las frecuencias de folículos y cuerpos lúteos disminuyeron vs hembras controles, valores que se restauraron luego de la superovulación. Sin embargo, en las hembras tratadas, la tasa de proliferación celular folicular y el desbalance de la expresión ovárica de VEGF (factor de crecimiento endotelial) persistieron luego de la superovulación. El número de ovocitos ovulados con metafase II anormal, fragmentados y activados partenogenéticamente fue mayor en las hembras tratadas respecto las controles. En conclusión, el consumo semicrónico moderado de alcohol produce anestro, ciclo estral irregular, foliculogénesis deficiente y anomalías núcleo-citoplasmáticas en los oocitos ovulados. Estas alteraciones podrían constituirse en un factor etiológico de pérdida gestacional temprana y desarrollo embrionario anormal luego del consumo de alcohol.

Abstract Chronic alcohol consumption is a global health problem that particularly affects the female population. However, the ef-fects of semi-chronic ethanol intake in low-moderate amounts on the ovary and oocyte are poorly understood. In a mouse model, 10% ethanol was administered in drinking water (treated females) or water (control females) for 15 days, and after superovulation or not (spontaneous ovulation), the estrous cycle and ovarian-gametic quality were analyzed. In treated females, the frequency and duration of the diestrus increased, and the frequencies of follicles and corpus luteum decreased vs control females, values that restored after superovulation. However, in treated females, the follicular cell proliferation rate and the imbalance in ovarian expression of VEGF (endothelial growth factor) persisted after superovulation. The number of ovulated oocytes with abnormal metaphase II, fragmented and parthenogenetically activated was higher in treated females than in control ones. In conclusion, moderate semi-chronic alcohol consumption produces anestrum, irregular estrous cycle, poor folliculogenesis, and nuclear-cytoplasmic abnormalities in ovulated oocytes. These alterations could constitute an etiological factor of early gestational loss and abnormal embryonic development after alcohol consumption.

Quantification of multi-oocyte follicles in ovaries of bitches / [Quantificação de folículos multiócitos em ovários de cadelas]

The objective of this study was to quantify the number and frequency of monocyte (MnOF) and multi-oocyte (MtOF) follicles in ovaries of bitches subjected to ovary salpingohysterectomy (OSH). Right and left ovaries of 38 bitches were collected after OSH, prepared, and a histological analysis was carried out. The ovaries were subjected to surface and deep histological cuts; the follicles were classified, and the number of follicles and cumulus oophorus complexes (COC) per follicle were quantified for each histological cut. MnOF and MtOF were found in all ovaries, at different developmental stages; primary follicles were grouped in the ovarian cortex, and follicles at other follicular stages presented a random distribution. MtOF containing two, three, four, or more COC were found in the ovaries of bitches, with a decreasing frequency trend, according to the number of COC in the MtOF. The effect of the age, number of estrus, estrus interval, and number of progenies per delivery was not significant for the number and frequency of MtOF in the ovaries of the bitches, whereas the size, number of pregnancies, use and number of contraceptive applications had some effect on the number and frequency of MtOF in the ovaries of the bitches.(AU)

Objetivou-se, com este estudo, quantificar o número e a frequência de folículos monocitários (MOF) e polioocitários (POF) provenientes de ovários de cadelas submetidas à ovariossalpingo-histerectomia (OSH). Para tanto, coletaram-se os ovários (direito e esquerdo) de 38 cadelas após OSH, com posterior preparação e análise histológica. Cada ovário foi submetido a dois cortes histológicos (superficial e profundo) onde se quantificou o número e a classificação dos folículos, bem como o número de complexos cumulus oophorus (COCs) por folículo em cada corte histológico. Observaram-se MOF e POF em todos os ovários estudados, em diferentes estádios de desenvolvimento, sendo os folículos primários agrupados no córtex ovariano, frente a uma distribuição aleatória dos outros estádios foliculares. FOPs contendo dois, três, quatro ou mais COCs foram observados nos ovários de todas as fêmeas estudadas, e sua frequência tendeu a diminuir de acordo com o número de COC presente no POF. Não se observou influência da idade, do número e do intervalo de estros, assim como do número de filhotes por gestação sobre o número/frequência de FOP nos ovários das cadelas estudadas, enquanto o porte, o número de gestações, o uso e o número de contraceptivo apresentaram algum grau de influência sobre o número/frequência de FOP nos ovários das cadelas estudadas.(AU)

Intravaginal progesterone device (1. 9g) and estradiol benzoate for follicular control in the mare during spring and summer / [Implante intravaginal de progesterona (1, 9g) e benzoato de estradiol no controle folicular de éguas durante a primavera e o verão]

The objective of this study was to evaluate follicular growth and ovulatory rates in mares treated with an intravaginal progesterone device (P4) during the 10-day period, associated with the use of estradiol benzoate (EB). The results were compared during the transition period (ET) in the spring and the breeding season in the summer (ER). The variables were submitted to ANOVA (Tukey's test), considering P<0.05. No ovulation occurred during the permanence of the P4 implant in both experimental periods. The ovulatory rate in the ER was 100% (n = 8) and in the ET 62.5% (n = 5; P = 0.0547). Significant differences were observed (<0.001), in both periods, comparing follicular growth rates during the permanence of P4 device (ER: 1.33 ± 0.89mm/d; ET: 1.00 ± 0.81mm/d) to the period without P4 (ER: 3.63 ± 1.33 mm/d; ET: 3.31 ± 1.66 mm/d). The present study demonstrated applicability and efficiency of a hormonal protocol using P4 intravaginal device and EB for follicular control in mares, both during ET and ER.

O objetivo deste trabalho foi avaliar a taxa de crescimento folicular e a taxa ovulatória em éguas tratadas com dispositivo intravaginal de progesterona (P4) durante o período de 10 dias, associado à utilização de benzoato de estradiol (BE). Os resultados foram comparados durante o período de transição (ET) da primavera com a época de reprodução no verão (ER). As variáveis foram submetidas à ANOVA (teste de Tukey), considerando-se P<0,05. Nenhuma ovulação ocorreu durante a permanência do dispositivo de P4 em ambos os períodos experimentais. A taxa ovulatória na ER foi de 100% (n = 8) e na ET, de 62,5% (n=5; P=0,0547). Diferença significativas (<0,001) foram observadas, em ambos os períodos experimentais, comparando as taxas de crescimento folicular durante a permanência da P4 (ER: 1,33 ± 0,89mm/d; ET: 1,00 ± 0,81mm/d) com o período sem P4 (ER: 3,63 ± 1,33mm/d; ET: 3,31 ± 1,66mm/d). O presente estudo demonstrou aplicabilidade e eficiência do protocolo hormonal utilizando dispositivo intravaginal de P4 e BE para controle folicular de éguas, tanto na ET quanto na ER.

Morphology and Biochemistry of Ovulation Morfologia e bioquímica da ovulação

Abstract The process of ovulation involves multiple and iterrelated genetic, biochemical, and morphological events: cessation of the proliferation of granulosa cells, resumption of oocyte meiosis, expansion of cumulus cell-oocyte complexes, digestion of the follicle wall, and extrusion of the metaphase-II oocyte. The present narrative review examines these interrelated steps in detail. The combined or isolated roles of the folliclestimulating hormone (FSH) and luteinizing hormone (LH) are highlighted. Genes indiced by the FSH genes are relevant in the cumulus expansion, and LH-induced genes are critical for the resumption ofmeiosis and digestion of the follicle wall. A nonhuman model for follicle-wall digestion and oocyte release was provided.

Resumo O processo de ovulação envolve modificações genéticas, bioquímicas e morfológicas múltiplas e interrelacionadas: suspensão da proliferação das células da granulosa, reinício da meiose do oócito, expansão das células do complexo cumulus-oócito, digestão da parede folicular, e extrusão do oócito. Esta revisão narrativa examina em detalhes cada um desses eventos e os principais genes e proteínas envolvidos. Mais importante, a ação combinada ou isolada do hormônio folículo-estimulante (HFE) e do hormônio luteinizante (HL) é destacada. Detalha-se o papel do HFE na expansão do cumulus e do HL na digestão da parede folicular, permitindo a extrusão do oócito na superfície ovariana. Proveu-se um modelo não humano para explicar a digestão da parede folicular.

Grado de desarrollo del folículo preovulatorio y su relación con el tamaño del cuerpo lúteo y la producción de progesterona en las razas Romosinuano (RS) y Brahman (Br) / Degree of development of the preovulatory follicle and its relationship with corpus luteum size and progesterone production in the Romosinuano (RS) and Brahman (Br) breeds

RESUMEN Para comparar en los folículos preovulatorios de hembras bovinas de las razas Brahman (Br) y Romosinuano (RS) el desarrollo de los diferentes componentes foliculares, como el tamaño del antro folicular, el grosor y el volumen ocupado por la capa de células de la granulosa (CG) y de la teca interna (TI), y su posible relación con el desarrollo del cuerpo lúteo (CL) y la producción de progesterona P4, se utilizaron 5 hembras de la raza RS y 5 de la raza Br. A estas se les realizó seguimiento ecográfico durante 2 ciclos estrales y se les efectuó muestreo de suero sanguíneo para determinar por Elisa los niveles de P4. En el tercer estro se les extirpó el ovario que contenía el folículo preovulatorio y se realizó un corte diametral sobre el estigma del folículo para seccionarlo en 2 partes iguales. En una de las partes se hicieron cortes histológicos y se determinó la morfometría folicular. La duración del ciclo estral en las hembras de la raza Br osciló entre 18 y 21 días, con promedio de 19,9 ± 1,6 días, y en las vacas RS entre 18 y 24 días, con un promedio 21,2 ± 1,69 días. La duración del estro fue de 13,9 ± 6,98 y 9,60 ± 4,72 h para las razas RS y Br, respectivamente. El intervalo estro-ovulación fue de 21,20 ± 5,07 h para los animales de la raza Br y de 24,40 ± 6,43 para los RS. No se registraron diferencias entre razas ni en el grosor (RS: 55,12 ± 6,46 μm vs. Br: 49,48 ± 17,07 μm), p > 0,05, tampoco en el volumen ocupado por la capa de CG (RS: 27,93 ± 6,11 mm1 vs. Br: 25,40 ± 13,85 mm3) de los folículos preovulatorios p < 0,05, en el grosor (RS: 122,50 ± 20,53 μm vs. Br: 129,61 ± 84,85 μm) o en el volumen (RS: 64,97 ± 19,71 mm1 vs. Br: 59,83 ± 25,67 mm3) de las células de la TI. El máximo desarrollo de los CL para la totalidad de las hembras se alcanzó el día 12,6 ± 4,05 (día 0 = estro), con un diámetro promedio de 22,92 ± 3,60 mm. Para las de la raza RS el diámetro máximo fue 23,06 ± 3,9 mm y se observó en promedio el día 14,88 ± 3,4. Para las de la raza Br fue el día 10,00 ± 3,16 con un promedio de 22,75 ± 3,16 mm. En ese día, la concentración media de P4 fue de 5,37 ± 1,38 ng/ml para la raza RS y 5,74 ± 0,89 ng/ml para la raza Br. No se presentaron diferencias significativas entre razas (p > 0,05). Se concluyó que no existen diferencias en los eventos fisiológicos estudiados entre las razas RS y Br. Los hallazgos del presente estudio, sin duda, pueden servir de base para futuros análisis en los bovinos residentes en el trópico.

ABSTRACT To compare in the preovulatory follicles of bovine females of the Brahman (Br) and Romosinuano (RS) breeds the development of the different follicular components, such as the size of the follicular antrum, the thickness and volume occupied by the granulosa (CG) and theca interna cells (TI) layers and their possible relationships with the development of the corpus luteum (CL) and progesterone (P4) production, 5 RS and 5 Br females were used; ultrasound follow-up was performed during 2 estrous cycles and blood serum sampling was carried out to determine progesterone (P4) levels by Elisa, at the third estrus, the ovary containing the preovulatory follicle was removed, and a diametral cut was made on the stigma of the follicle to divide it into 2 equal parts. Histological sections were made of one of the parts and follicular morphometry was determined. Duration of the estrous cycle in Br females ranged between 18 and 21 days, with an average of 19,9 ± 1,6 days and between 18 and 24 with an average of 21,2 ± 1,69 days for RS cows. Estrus duration was 13,9 ± 6,98 and 9,60 ± 4,72 h for RS and Br breeds, respectively. The estrus-ovulation interval was 21,20 ± 5,07 h for the Br breed animals and 24,40 ± 6,43 RS. There were no differences between breeds or in thickness (RS: 55,12 ± 6,46 μm vs. Br: 49,48 ± 17,07 μm) p > 0,05, nor in the volume occupied by the granulosa cells layer (RS: 27,93 ± 6,11 mm3 vs. Br: 25,40 ± 13,85 mm3) of pre-ovulatory follicles; neither in the thickness (RS: 122,50 ± 20,53 vs. Br: 129,61 ± 84,85 μm) nor in the volume (RS: 64,97 ± 19,71 mm3 vs. Br: 59,83 ± 25,67 mm3) of the internal theca cells; p < 0,05. The maximum CL development for all the females was observed on day 12,6 ± 4,05 (day 0 = estrus), with an average diameter of 22,92 ± 3,60 mm. For those of the RS breed the maximum diameter was observed on average on day 14,88 ± 3,4, with 23,06 ± 3,9 mm, while for those of the Br breed it was day 10,00 ± 3,16 with an average diameter of 22,75 ± 3,16 mm; at this time the mean progesterone concentration was 5,37 ± 1,38 ng/ml for the RS breed and 5,74 ± 0,89 ng/ml for the Br breed. There were no significant differences between breeds (p-value: 0,5561). It was concluded that there are no differences in the physiological events studied between the studied breeds. Present results, undoubtedly, might serve as basis to future studies in bovines in tropical zones.

Relationship between superovulation and embryo production with ovarian follicular population before superovulatory treatment in Brazilian Bergamasca sheep / [Relação da resposta superovulatória e produção embrionária com a população folicular ovariana antes do tratamento superovulatório de ovelhas da raça Bergamácia Brasileira]

The objective of this study was to quantify the superovulatory response and embryo production of Brazilian Bergamasca sheep and to evaluate the link to the follicular condition before superovulatory treatment, as a reference for selection of donors with potential for superovulation. Follicular population of twenty-three sheep was evaluated by ultrasound during metestrus phase of the estrous cycle and divided into groups of low, medium and high follicular population. Subsequently, they were synchronized, superovulated with 133mg of pFSH, mated and subjected to embryo collection. The superovulatory response (9.0±3.3 vs 10.7±6.2 vs 13.8±7.1) and embryo production (4.0±3.8 vs 2.6±2.0 vs 1,8±4.0) were similar between groups (P>0.05). There was a positive correlation between the number of follicles during the metestrus phase and the number of corpus luteum with premature regression (PLR) (0.52) and a negative correlation between the recovery rate and PLR (-0.44) (P<0.05). The sheep that presented PLR had more follicles during metestrus (16.9±7.8 vs 12.7±3.2) and lower embryo recovery rate (38.8±29.3 vs 72.2±29.9) than those with functional CLs (P<0.05). Follicular quantification during metestrus phases was unable to identify donors with high embryo production. Animals with PLR had a larger follicular population during metestrus and lower embryo recovery rate.(AU)

O objetivo deste trabalho foi quantificar a resposta superovulatória e a produção embrionária de ovelhas Bergamácia Brasileira e relacioná-las com a condição folicular antes do tratamento superovulatório, como referência para seleção de doadoras com potencial para superovulação. Vinte e três ovelhas foram avaliadas quanto à população folicular por ultrassonografia na fase de metaestro do ciclo estral e divididas em grupos com baixa, média e alta população folicular. Posteriormente foram sincronizadas, superovuladas com 133mg de pFSH, acasaladas e submetidas à coleta de embriões. A resposta superovulatória (9,0±3,3 vs. 10,7±6,2 vs. 13,8±7,1) e a produção embrionária (4,0±3,8 vs. 2,6±2,0 vs. 1,8±4,0) foram semelhantes entre os grupos (P>0,05). Houve correlação positiva entre o número de folículos no metaestro e o número de corpos lúteos com regressão prematura (RPCL) (0,52) e correlação negativa entre a taxa de recuperação e RPCL (-0,44) (P <0,05). As ovelhas que apresentaram RPCL tiveram mais folículos no metaestro (16,9±7,8 vs. 12,7±3,2) e menor taxa de recuperação embrionária (38,8±29,3 vs. 72,2±29,9) do que as que apresentaram CLs funcionais (P<0,05). A quantificação folicular nas fases de metaestro não foi capaz de identificar doadoras com alto potencial de produção embrionária. Animais com RPCL tiveram maior população folicular no metaestro e menor recuperação de embriões.(AU)

Effects of bisphenol A on apoptosis of ovarian preantral follicular granulosa cells and ovarian development in mice / 南方医科大学学报

OBJECTIVE@#To investigate the effect of environmental estrogen bisphenol A (BPA) exposure on apoptosis of mouse ovarian preantral follicular granulosa cells and ovarian development and explore the underlying mechanism.@*METHODS@#Mouse ovarian preantral follicular granulosa cells were isolated from female ICR mice at postnatal day (PND) 10 and cultured @*RESULTS@#Compared with the control cells group, the isolated cells exposed to a low concentration of BPA (50 μmol/L) showed a significantly lowered apoptosis rate, increased mitochondrial membrane potential, and enhanced cellular proliferation (@*CONCLUSIONS@#BPA can concentration-dependently regulate the function of ovarian preantral follicular granulosa cells in mice and potentially affects both the pregnant mice and the offspring female mice in light of early ovarian development.

Histone methylation in mammalian follicular development / 生理学报

The normal development of follicles involves a series of complex life processes such as ordered transcriptional activation and inhibition, which is crucial for female reproductive ability. Histone methylation can change the chromatin state in cells and affect the transcription activity of genes. Current studies indicate that epigenetic modifications such as histone methylation play an important regulatory role in follicular development in female mammals. This paper summarized the relationship between H3K4, H3K9 methylation and germ cell development, their regulatory effects, including their dynamical changes during follicular development, and the progress of H3K4me3 and other histone methylation binding to promoter regions of different genes to regulate gene expression and thus affect germ cell epigenetic reprogramming, oocyte transcription, meiosis and other processes. This review will provide a reference for the study of mechanisms related to histone methylation modification and the development and maturation of gonadal parenchymal cells.

Advances of the Regulation of microRNAs in Follicular Development / 中国医学科学院学报

In recent years,microRNAs(miRNAs)have been detected at different stages of follicular development and in different cells of follicles.Extracellular vesicle(EV)-derived miRNAs have also been detected in the follicular fluid of mature follicles.miRNAs participate in the regulation of normal follicular development,and the regulation disorder may lead to the occurrence of some ovarian diseases.In order to further systematically elucidate the regulatory mechanism of miRNAs on follicular development and find suitable EV-derived miRNAs that can predict oocyte development,we reviewed the functions of miRNAs in follicular development from the perspectives of granulosa cell development,oocyte development,and hormone synthesis.

Modified Dihuang Decoction improves ovarian reserve in mice by regulating Bcl-2-related mitochondrial apoptosis pathway / 中国中药杂志

The present study investigated the effect of Modified Dihuang Decoction in improving ovarian reserve in mice through the Bcl-2-related mitochondrial apoptosis pathway. Forty-eight adult female BALB/c mice were randomly divided into the following six groups with eight mice in each group: a blank group, a model group, a femoston group(three cycles of treatment with 0.13 mg·kg~(-1) estradiol tablets for 2 days and 1.43 mg·kg~(-1) estradiol and dydrogesterone tablets for 3 days), and high(64.74 g·kg~(-1))-, medium(43.16 g·kg~(-1))-, and low-dose(21.58 g·kg~(-1)) Modified Dihuang Decoction groups. Mice in other groups except the blank group received a single intraperitoneal injection of 12 mg·kg~(-1) cyclophosphamide and 1.2 mg·kg~(-1) busulfan to induce a model of diminished ovarian reserve(DOR), while those in the blank group received an equal volume of normal saline. Mice were treated with corresponding drugs for 15 d from the 36 th day, once per day, and the mice in the blank group and the model group were treated with an equal volume of normal saline. The general condition and oestrous cycle were observed. The serum hormone levels were detected with the enzyme-linked immunosorbent assay(ELISA). The morphological changes of ovaries were observed by HE staining. Western blot was used to detect the protein expression of cysteinyl aspartate specific proteinase-9(caspase-9), cleaved caspase-3, Bcl-2 associated X protein(Bax), Bcl-2, superoxide dismutase-2(SOD-2), and glutathione peroxidase-1(GPx-1). The mRNA expression of Bax and Bcl-2 was detected by real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR). The results showed that compared with the blank group, the model group showed body weight loss, disordered oestrous cycle, elevated serum levels of follicle-stimulating hormone(FSH) and luteinizing hormone(LH), reduced serum levels of estradiol(E_2), anti-mullerian hormone(AMH), and inhibin B(INHB), the declining number of ovarian follicles and granulosa layers, increased number of atretic follicles, up-regulated protein expression of caspase-9, cleaved caspase-3, and Bax and Bax mRNA expression in ovaries, and down-regulated protein expression of Bcl-2, SOD-2 and GPx-1, and Bcl-2 mRNA expression. Compared with the model group, the Modified Dihuang Decoction groups displayed restored body weight and oestrous cycle, decreased serum levels of FSH and LH, elevated serum levels of E_2, AMH, and INHB, increased number of ovarian follicles, thickened granulosa layers, and declining number of atretic follicles. Additionally, the protein expression of caspase-9, cleaved caspase-3, and Bax, and Bax mRNA expression was down-regulated, and the protein expression of Bcl-2, SOD-2, and GPx-1, and Bcl-2 mRNA expression was up-regulated. The results suggest that Modified Dihuang Decoction can regulate endocrine hormone, promote follicle growth and improve ovarian reserve by enhancing ovarian anti-oxidant capacity, inhibiting the Bcl-2-related mitochondrial apoptosis pathway, and further inhibiting cell apoptosis.

Effects of Vitrification with Self-made Carriers and Slow Programmed Freezing on Ovarian Tissue of Sheep / 中国医学科学院学报

Objective To investigate the effects of self-made carriers on the cryopreservation of ovarian tissue of sheep. Methods Thirty-two ovaries were randomly assigned to fresh group,programmed freezing group,self-made carrier I vitrification group,and self-made carrier Ⅱ vitrification group.The morphology,proliferation,apoptosis,and estrogen level of the ovarian tissue in each group were observed. Results After cryopreservation,the morphology normal rate of the primordial follicles in programmed freezing group,self-made carrier I vitrification group,and self-made carrier Ⅱ vitrification group were 74.2%,72.8%,and 72.3%,respectively,lower than that(83.7%)in the fresh group(χ