Perilipin1 inhibits Nosema bombycis proliferation by promoting Domeless- and Hop-mediated JAK-STAT pathway activation in Bombyx mori.

Lipid droplets (LDs) are dynamic organelles that participate in the regulation of lipid metabolism and cellular homeostasis inside of cells. LD-associated proteins, also known as perilipins (PLINs), are a family of proteins found on the surface of LDs that regulate lipid metabolism, immunity, and other functions. In silkworms, pébrine disease caused by infection by the microsporidian Nosema bombycis (Nb) is a severe threat to the sericultural industry. Although we found that Nb relies on lipids from silkworms to facilitate its proliferation, the relationship between PLINs and Nb proliferation remains unknown. Here, we found Nb infection caused the accumulation of LDs in the fat bodies of silkworm larvae. The characterized perilipin1 gene (plin1) promotes the accumulation of intracellular LDs and is involved in Nb proliferation. plin1 is similar to perilipin1 in humans and is conserved in all insects. The expression of plin1 was mostly enriched in the fat body rather than in other tissues. Knockdown of plin1 enhanced Nb proliferation, whereas overexpression of plin1 inhibited its proliferation. Furthermore, we confirmed that plin1 increased the expression of the Domeless and Hop in the JAK-STAT immune pathway and inhibited Nb proliferation. Taken together, our current findings demonstrate that plin1 inhibits Nb proliferation by promoting the JAK-STAT pathway through increased expression of Domeless and Hop. This study provides new insights into the complicated connections among microsporidia pathogens, LD surface proteins, and insect immunity.IMPORTANCELipid droplets (LDs) are lipid storage sites in cells and are present in almost all animals. Many studies have found that LDs may play a role in host resistance to pathogens and are closely related to innate immunity. The present study found that a surface protein of insect lipid droplets could not only regulate the morphological changes of lipid droplets but also inhibit the proliferation of a microsporidian pathogen Nosema bombycis (Nb) by activating the JAK-STAT signaling pathway. This is the first discovery of the relationship between microsporidian pathogen and insect lipid surface protein perilipin and insect immunity.

Resveratrol reverses Palmitic Acid-induced cow neutrophils apoptosis through shifting glucose metabolism into lipid metabolism via Cav-1/ CPT 1-mediated FAO enhancement.

Elevated plasma nonesterified fatty acids (NEFAs) affect neutrophils function and longevity during the periparturient period in dairy cows. Previous research has shown that resveratrol (RSV) may protect cell viability from NEFA-induced damage by regulating energy metabolism. However, it is unclear whether RSV has a protective effect on palmitic acid (PA)-treated neutrophils. The aim of this study was to investigate the molecular regulatory mechanism of the protective effect of RSV on neutrophils. The results showed that treatment with high concentrations of RSV (50 µM, 100 µM) maintained neutrophils activity by inhibiting neutrophils apoptosis (P < 0.05). Further analysis showed that high concentrations of RSV enhanced fatty acid oxidation (FAO) to produce ATP by promoting the expression of CAV1, ACSL-1 and CPT1 (P < 0. 05) while inhibiting glycolysis by suppressing PFK1 activity (P < 0. 05) and reducing glucose transport-related protein (GLUT1/GLUT4) expression by inhibiting glucose uptake (P < 0.05). These results suggest that RSV protects neutrophils from PA-induced apoptosis by regulating energy metabolism. Our results revealed that RSV protects neutrophils from PA-induced apoptosis by shifting glucose metabolism to lipid metabolism. This study tenders to a meaningful understanding of the effects of RSV on neutrophils function in periparturient cows suffering from negative energy balance (NEB).

Dexamethasone enhances glucose uptake by SGLT1 and GLUT1 and boosts ATP generation through the PPP-TCA cycle in bovine neutrophils.

BACKGROUND: Clinical dexamethasone (DEX) treatment or stress in bovines results in extensive physiological changes with prominent hyperglycemia and neutrophils dysfunction. OBJECTIVES: To elucidate the effects of DEX treatment in vivo on cellular energy status and the underlying mechanism in circulating neutrophils. METHODS: We selected eight-month-old male bovines and injected DEX for 3 consecutive days (1 time/d). The levels of glucose, total protein (TP), total cholesterol (TC), and the proinflammatory cytokines interleukin (IL)-1ß, IL-6 and tumor necrosis factor (TNF)-α in blood were examined, and we then detected glycogen and adenosine triphosphate (ATP) content, phosphofructosekinase-1 (PFK1) and glucose-6-phosphate dehydrogenase (G6PDH) activity, glucose transporter (GLUT)1, GLUT4, sodium/glucose cotransporter (SGLT)1 and citrate synthase (CS) protein expression and autophagy levels in circulating neutrophils. RESULTS: DEX injection markedly increased blood glucose, TP and TC levels, the Ca2+/P5+ ratio and the neutrophil/lymphocyte ratio and significantly decreased blood IL-1ß, IL-6 and TNF-α levels. Particularly in neutrophils, DEX injection inhibited p65-NFκB activation and elevated glycogen and ATP contents and SGLT1, GLUT1 and GR expression while inhibiting PFK1 activity, enhancing G6PDH activity and CS expression and lowering cell autophagy levels. CONCLUSIONS: DEX induced neutrophils glucose uptake by enhancing SGLT1 and GLUT1 expression and the transformation of energy metabolism from glycolysis to pentose phosphate pathway (PPP)-tricarboxylic acid (TCA) cycle. This finding gives us a new perspective on deeper understanding of clinical anti-inflammatory effects of DEX on bovine.

Acceptance of COVID-19 vaccination and influencing factors among people living with HIV in Guangxi, China: a cross-sectional survey.

BACKGROUND: Vaccination has been proven to be an effective approach against the coronavirus disease 2019 (COVID-19) pandemic. This study aimed to determine the acceptance rate and factors influencing acceptance of COVID-19 vaccination among people living with HIV (PLWH) in Guangxi, China. METHODS: A cross-sectional survey was carried out in five cities in Guangxi, China from May 7 to June 1, 2021. Questionnaires on the acceptance of COVID-19 vaccination and the related factors were conducted among PLWH recruited by simple random sampling. Univariate and multivariate logistic regression analyses were performed to identify factors associated with acceptance of COVID-19 vaccination. RESULTS: Of all valid respondents (n = 903), 72.9% (n = 658) were willing to receive COVID-19 vaccination. Fear of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection was the main reason for being willing to receive vaccination (76.0%), while the main reasons for not willing were the concerns about vaccine safety (54.7%) and the vaccination's effect on antiretroviral therapy (ART) (50.6%). The most important factors influencing acceptance were the perception that vaccination is unsafe for HIV-infected people (aOR = 0.082, 95% CI = 0.024-0.282) and the poor efficacy in preventing SARS-CoV-2 infection in HIV-infected people (aOR = 0.093, 95% CI = 0.030-0.287). Other factors associated with acceptance included Zhuang ethnicity (aOR = 1.653, 95% CI = 1.109-2.465), highest education level of middle school, high school or above (aOR = 1.747, 95% CI = 1.170-2.608; aOR = 2.492, 95% CI = 1.326-4.682), and the vaccination having little effect on ART efficacy (aOR = 2.889, 95% CI = 1.378-6.059). CONCLUSIONS: Acceptance rate of the COVID-19 vaccination is relatively low among PLWH compared to the general population in China, although some patients refused vaccination due to concerns about vaccine safety and vaccination affecting ART efficacy. More research is needed to investigate the impact of the COVID-19 vaccines on ART efficacy and the effectiveness in preventing SARS-CoV-2 infection among PLWH.

[Relationship between Polymorphism of miR-155 and Its Target Gene MyD88 and Clinicopathological Features of Diffuse Large B-cell Lymphoma].

OBJECTIVE: To investigate the relationship between the polymorphism of miR-155 and its target gene MyD88 and clinicopathological features of diffuse large B-cell lymphoma (DLBCL). METHODS: 135 cases of DLBCL patients in our hospital from March 2015 to August 2017 were selected, and 90 cases of reactive hyperplasia of lymph nodes were selected as the control group. The relative expression of miR-155 and MyD88 gene polymorphism were detected in the two groups, and the relationship between miR-155 and MyD88 gene polymorphism and clinicopathological characteristics of DLBCL was analyzed. RESULTS: The relative expression of miR-155 in DLBCL patients was significantly higher than that in the control group (P<0.05). The mutation rate of MyD88 L265P in DLBCL group was significantly higher than that in control group (P<0.05). The relative expression of miR-155 in patients with MyD88 L265P mutation was significantly higher than that in patients with wild-type DLBCL (P<0.05). The relative expression of miR-155 and the polymorphism of MyD88 L265P were associated with lesion location, stage, BCL-2 protein expression and MyD88 protein expression in DLBCL patients (t=7.461、8.804、6.487、10.812； χ2=10.681、8.599、7.251、23.008；P<0.05). The survival of DLBCL patients with low miR-155 expression was significantly better than that with high miR-155 expression (P<0.05). The survival of wild-type DLBCL patients with MyD88 L265P locus was significantly better than that of mutant DLBCL patients (P<0.05). There was a significant positive correlation between the relative expression of miR-155 and the expression of MyD88 protein (r=0.428, P=0.000). CONCLUSION: The abnormal expression of miR-155 and the mutation rate of MyD88 gene in DLBCL patients are increased, and the expression of miR-155 and the mutation of MyD88 gene affect the disease progression and prognosis of patients, which may be potential biological indicators for the diagnosis, treatment and prognosis of DLBCL.

TUC338 Promotes Diffuse Large B Cell Lymphoma Growth via Regulating EGFR/PI3K/AKT Signaling Pathway.

TUC338 is emerging as a novel vital long noncoding RNA (lncRNA) in human cancer; however, its role in diffuse large B cell lymphoma (DLBCL) remains unknown. In this study, we found that TUC338 was remarkably upregulated in DLBCL tissues as compared to matched normal tissues. High TUC338 was closely related to advanced Ann Arbor stage, resistance to CHOP-like treatment, and high IPI (International Prognostic Index). Stable knockdown of TUC338 evidently inhibited cell proliferation and chemotherapy resistance to Adriamycin and induced apoptosis. Further, we found that TUC338 was able to directly bind to miR-28-5p and increased EGFR level, resulting in activating carcinogenic PI3K/AKT signaling, thereby facilitating DLBCL uncontrolled growth. Moreover, we also found that depletion of TUC338 led to the inactivation of EGFR/PI3K/AKT pathway in vivo by using the xenograft tumor model. Preclinically, DLBCL patients with high TUC338 had shorter survival time than those with low TUC338, and serum TUC338 level was identified as an excellent indicator for DLBCL diagnosis. In sum, our findings clearly indicate that TUC338 functions as an oncogenic lncRNA in DLBCL through activating EGFR/PI3K/AKT pathway via sponging and inhibiting miR-28-5p, which may be a promising target for DLBCL treatment.

[Effects of Long Non-coding RNA-TUC338 on the Migration and Proliferation of Lymphoma Cells via PI3K/AKT Signaling Pathway].

OBJECTIVE: To investigate the effect of long non-coding RNA-TUC338 on the proliferation and migration of lymphoma cells. METHODS: The expression of TUC338 in different lymphoma cells was detected by fluorescence quantitative PCR, cell proliferation by sulforhodamine B (SRB) assay, migration of lymphoma cells by transwell assay, and protein expression in PI3K/AKT signaling pathway by Western blot. RESULTS: The expression levels of TUC338 in lymphoma cells Daudi, U937, BC-3, and Raji significantly increased in comparison with human normal T lymphocytes H9 (t=13.277, 10.103, 16.200, and 26.687, P=0.002, 0.005, 0.001, and 0.000). Compared with NC-siRNA group, the number of cells crossing the chamber of TUC338-siRNA group was significantly reduced (t=30.508, P=0.000), the protein expression levels of p-PI3K and p-AKT significantly decreased (t=16.872 and 18.371, P=0.000 and 0.000), and OD530 absorbance values at 24 h, 48 h, and 72 h were significantly lower (P<0.05). CONCLUSION: The expression of TUC338 significantly increases in lymphoma cells, and silence of TUC338 effectively inhibits the activation of PI3K/AKT signaling pathway, thereby inhibiting the proliferation and migration of lymphoma cells, which has a potential application value in diagnosis and treatment of lymphoma.

Treatment of cattle oocytes with C-type natriuretic peptide before in vitro maturation enhances oocyte mitochondrial function.

The present study was conducted to evaluate the effects in vitro on oocyte mitochondrial function of C-type natriuretic peptide (CNP) when treatments were imposed before in vitro maturation (IVM). Immature oocytes were either directly matured in vitro for 24 h (Control, no pre-IVM), or cultured in basic medium not supplemented or supplemented with CNP (100 nM) (Control pre-IVM and CNP pre-IVM, respectively) for 6 h, followed by IVM for 24 h. The results indicated treatment with CNP before IVM affected patterns of distribution of mitochondria, increased the mitochondrial content, membrane potential, and decreased the ROS content in cattle oocytes before and after IVM. Furthermore, treatment of immature cattle oocytes with CNP before IVM induced marked increases in the relative abundance of mRNA transcripts and proteins related to mitochondria development and antioxidative defense mechanisms. Treatment with CNP before oocyte IVM also resulted in an enhanced relative abundance of sirtuin-1 (SIRT1) mRNA transcript in cattle oocytes. Taken together, these results provide evidence that treatment of cattle oocytes with CNP before IVM improved mitochondrial function and antioxidant defense mechanisms in cattle oocytes. Findings in the present study provide insights into the potential mechanisms by which CNP has positive effects on oocyte cytoplasmic organelles, specifically mitochondria.

Effects of C-type natriuretic peptide on meiotic arrest and developmental competence of bovine oocyte derived from small and medium follicles.

The present study aimed to evaluate the effects of C-type natriuretic peptide (CNP) on meiotic arrest and developmental competence of bovine oocyte derived from follicles of different sizes. Collected immature cumulus-oocyte complexes from small follicles (< 3 mm) and medium follicles (3-8 mm) were cultured for 6 h in basal medium supplementated without or with 200 nM CNP. We observed that CNP effectively sustained meiotic arrest at germinal vesicle stage in in vitro cultured bovine oocytes from follicles of different sizes. Moreover, CNP treatment significantly improved the levels of cGMP in both cumulus cells and oocytes, as well as the levels of cAMP in oocytes regardless of follicle size. Based on the above results, we tested the effect of a novel in vitro maturation (IVM) system based on CNP-pretreatment, including a pre-IVM phase for 6 h using 200 nM CNP, followed by a extended IVM phase for 28 h, on developmental competence of bovine oocyte derived from small follicles (< 3 mm) and medium follicles (3-8 mm) compared to standard IVM system. The results showed that athough the novel IVM system based on CNP-pretreatment enhanced the developmental potencial of oocytes obtained from large follicles, but had no effect on the developmental comptence of oocytes obtained from small follicles.

[Relationship of Expression of Circ_cgga162 with the Prognosis of Patients with Mantle Cell Lymphoma].

OBJECTIVE: To investigate the expression of Circ_cgga162 in serum of mantle cell lymphoma (MCL) patients and analyze its potential as a prognostic biomarker. METHODS: The expression of Circ_cgga162 in 86 cases of mantle cell lymphoma and 50 cases of lymph node reactive hyperplasia (RH) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). The relationship between the expression of Circ_cgga162 and clinicopathological features was analyzed by univariate analysis. The relationship of Circ_cgga162 expression with progression-free survival time and overall survival time was analyzed by Kaplan-Meier. The relationship between expression of Circ_cgga162 and prognosis of patients was analyzed by univariate and multivariate analysis. RESULTS: The expression level of Circ_cgga162 in MCL patients was significantly higher than that in control (RH) group (P＜0.01). The expression of Circ_cgga162 not correlated with age, gender, B symptoms and LDH (all P＞0.05), but correlated with the expression of MCL International Prognostic Index (IPI), Ann Arbor stage, bone marrow infiltration and Ki67 (all P＜0.05). In addition, Kaplan-Meier analysis showed that the progression-free survival time and overall survival time of the MCL patients with high expression of Circ_cgga162 were significantly shorter than those of the MCL patients with low expression (P＜0.01). Univariate analysis showed that Ann Arbor stage, Circ_cgga162 expression, MIPI, bone marrow infiltration and Ki67 were the prognostic factors for MCL patients (all P＜0.05). Multivariate Cox regression analysis showed that Ann Arbor stage, Circ_cgga162 expression and MIPI were independent factors affecting the prognosis of MCL patients (all P＜0.05). CONCLUSION: Circ_cgga162 is highly expressed in serum of patients MCL, which relates with the prognosis of MCL patients. Circ_cgga162 can be used as a potential prognostic marker and therapeutic target for MCL patients.

Strategies to approach high performance in Cr3+-doped phosphors for high-power NIR-LED light sources.

Broadband near-infrared (NIR)-emitting phosphors are key for next-generation smart NIR light sources based on blue LEDs. To achieve excellent NIR phosphors, we propose a strategy of enhancing the crystallinity, modifying the micromorphology, and maintaining the valence state of Cr3+ in Ca3Sc2Si3O12 garnet (CSSG). By adding fluxes and sintering in a reducing atmosphere, the internal quantum efficiency (IQE) is greatly enhanced to 92.3%. The optimized CSSG:6%Cr3+ exhibits excellent thermal stability. At 150 °C, 97.4% of the NIR emission at room temperature can be maintained. The fabricated NIR-LED device emits a high optical power of 109.9 mW at 520 mA. The performances of both the achieved phosphor and the NIR-LED are almost the best results until now. The mechanism for the optimization is investigated. An application of the NIR-LED light source is demonstrated.

Electron-phonon coupling mechanisms of broadband near-infrared emissions from Cr3+ in the Ca3Sc2Si3O12 garnet.

Cr3+ in the Ca3Sc2Si3O12 garnet (CSSG) has the ability to convert blue light to broadband near-infrared (NIR) emissions, which is a promising strategy for next-generation smart NIR light sources based on blue LEDs. The Cr3+ luminescence strongly depends on temperature due to electron-phonon coupling (EPC). We reveal the EPC mechanism of Cr3+ in CSSG for the first time by temperature-dependent photoluminescence measurement from 77 to 573 K and cathodoluminescence using a scanning electron microscope. Cr3+ occupies the Sc3+ site and experiences a weak crystal field in CSSG, manifesting a broad NIR emission in the 700-900 nm range that originates from the 4T2g→4A2g transition. The zero phonon line (ZPL) of the 4T2 state is observed at ∼713 nm with a vibrational energy of ∼310 cm-1. A strong EPC leads to a large Stokes shift (∼2900 cm-1). The Huang-Rhys parameter (S = 4), crystal field strength (Dq/B), and Racah parameters (B and C) are estimated.

[Expression and Clinical Significance of LncRNA KCNQ1OT1 in Patients with Acute Myeloid Leukemia].

OBJECTIVE: To investigate the expression of LncRNA KCNQ1OT1 in patients with acute myeloid leukemia (AML) and to analyze the relation of LncRNA KCNQ1OT1 expression levels with clinicopathological features. METHODS: A total of 68 patients with AML were enrolled in the study, 48 out of them were suffered from acute myeloid leukemia (AML) and 20 reached to complete remission (CR), 30 age-matched patients with iron-deficient anemia were included in control group, the peripheral blood samples of all the patients were collected, and the real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression of LncRNA KCNQ1OT1, meanwhile, the correlation of its expression with clinicopathological characteristics and prognosis was analyzed. RESULTS: The expression of LncRNA KCNQ1OT1 in AML patients was significantly higher than that in the patient with complete remission and iron-deficient anemia (F=14.67, P<0.01). The expression of LncRNA KCNQ1OT1 was not significantly different between 20 cases of AML-CR and 30 cases of iron-deficient anemia (P>0.05). The expression of LncRNA KCNQ1OT1 was associated with NCCN risk grade and survival status in patients with AML. The median overall survival time was significantly shorter in patients with high expression of LncRNA KCNQ1OT1 than that in patients with low expression(P<0.05). CONCLUSION: LncRNA KCNQ1OT1 may be involved in the regulation of AML. Expression of LncRNA KCNQ1OT1 and NCCN risk score can be used as biomarkers of prognosis in the patients with AML and may be a potential prognostic marker and therapeutic target for AML patients.

Natriuretic peptide receptor 2 (NPR2) localized in bovine oocyte underlies a unique mechanism for C-type natriuretic peptide (CNP)-induced meiotic arrest.

Meiosis is of prime importance for successful gametogenesis, and insufficient maintenance of oocyte meiotic arrest compromises oocyte developmental competence. Recent studies have demonstrated that the C-type natriuretic peptide (CNP)-Natriuretic peptide receptor 2 (NPR2) pathway can inhibit mammalian oocyte meiotic resumption. In mouse and porcine, the inhibitory effect of mural granulosa cell (MGC)-derived CNP on oocyte meiotic resumption is mediated by NPR2 localized in cumulus cells (CCs) surrounding the oocytes. However, in the present study, we identified a novel mechanism for CNP-induced meiotic arrest that appears to be unique to bovine oocytes. Unlike mouse and porcine, bovine NPR2 not only localizes in CCs, but also in oocyte membranes. We also showed that CNP can directly activate intra-oocyte cGMP production via NPR2 localized in oocyte membranes, in parallel with the CC-mediated pathway. Furthermore, we demonstrated that Npr2 expression in bovine CCs and oocytes were synergistically regulated by estradiol and oocyte-derived growth factors. Finally, based on the profound inhibitory effect of CNP on meiotic resumption, we established a natural factor synchronized in vitro oocyte maturation (NFSOM) system, which can significantly improve the developmental competence of matured oocytes, thereby resulting in higher in vitro embryo production efficiency. Taken together, our study not only provides new insight into understanding the crosstalk between oocytes and follicular somatic cells in mammals, but also presents a promising strategy for improving the in vitro oocyte maturation systems of assisted reproductive technology (ART).

Mitochondria and pluripotent stem cells function.

Mitochondria are important intracellular organelles which provide energy for cellular activities through oxidative phosphorylation. Recently, mitochondria have been shown to exhibit peculiar features in pluripotent stem cells (PSCs), namely, PSCs rely mainly on glycolysis for energy supply in pluripotent states while mitochondrial oxidative phosphorylation function is gradually enhanced during PSCs differentiation. In contrast, during somatic reprogramming, the metabolic transition from mitochondrial oxidative phosphorylation to glycolysis is necessary for successful reprogramming. Moreover, mitochondrial biogenesis and dynamics are also actively involved in the maintenance of pluripotency, induction of differentiation and induced pluripotent stem cells (iPSCs) reprogramming. Here we reviewed mitochondrial structure and function in regulating PSCs pluripotency, anabolism, redox homeostasis, differentiation, and reprogramming, which may provide reference for further understanding the role of mitochondria in PSCs.

Mechanisms of TET protein-mediated DNA demethylation and its role in the regulation of mouse development.

TET (ten-eleven translocation) protein family includes three members TET1, TET2 and TET3, which belong to alpha-ketoglutaric acid ( α-KG )- and Fe(2+)-dependent dioxygenase superfamily, and have the capacity to convert 5-methylcytosine (5 mC) to 5-hydroxymethylcytosine (5 hmC), 5-formylcytosine (5 fC) and 5-carboxylcytosine (5 caC). At present, growing lines of evidence indicate that TET proteins are involved in the control of active or passive DNA demethylation via different mechanisms; moreover, their activities may be regulated by some cellular factors. TET proteins play vital roles in modulating mammal development, including primordial germ cell formation, embryonic development, stem cells pluripotency, nerve and brain development, etc. The identification of biological roles of TET proteins will open a new field in epigenetic research, and these studies on TET proteins are of great significance to life science research. Here, we review TET proteins from their structure, molecular mechanisms of DNA demethylation and function in the regulation of mouse development, which may provide the basis for understanding the functions of TET proteins.

Effects of administration of estradiol and progesterone on ovarian stimulation response, and in vitro developmental competence of oocytes in CIDR-treated calves.

The present study was conducted to investigate the effects of administering estradiol (estradiol-17ß, 2.5 mg) plus progesterone (P4, 50 mg) as a part of a controlled internal drug release (CIDR)-based superovulation protocol on ovarian follicles development, and oocyte recovery as well as on in vitro development in prepubertal calves after follicle stimulating hormone (FSH) stimulation. Calves in the treatment group exhibited significantly decreased (P < 0.05) numbers of both small and large antral follicles compared to the control group A and B (<3 mm in diameter [small], 3.4 versus 12.8, 11.6; > 7 mm in diameter [large], 0 versus 7.6, 7.2). The treated animals also exhibited an increased (P < 0.05) proportion of usable oocytes from the total oocytes recovered compared with those of the control group A and B (77% versus 50.2%, 47.8%). Moreover, the rate of cleavage and the percentage of blastocysts displayed an increased trend in the treatment group compared to controls A and B (59.2% versus 50.5%, 50.8%; 15% versus 12.4%, 11.6%, respectively). In conclusion, the application of estradiol-17ß plus P4 at CIDR insertion prior to FSH stimulation affected ovarian follicles development and exerted a beneficial effect on the in vitro development of calf oocytes.