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1.
J Cell Physiol ; 239(1): 166-179, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37991438

RESUMEN

Abnormal function and fibrosis of endometrium caused by cows' endometritis pose difficult implantation of embryos and uterine cavity adhesions. 17ß-Estradiol (E2) serves as the most effective aromatized estrogen, and its synthetase and receptors have been detected in the endometrium. Studies have demonstrated the positive role of estrogen in combating pathological fibrosis in diverse diseases. However, it is still unknown whether E2 regulates endometrium fibrosis in bovine endometritis. Herein, we evaluated the expression patterns of transforming growth factor-ß1 (TGF-ß1), epithelial-mesenchymal transformation (EMT)-related proteins (α-SMA, vimentin N-cadherin and E-cadherin), cytochrome P450 19A1 (CYP19A1), and G protein-coupled estrogen receptor (GPER) in bovine healthy endometrium and Inflammatory endometrium. Our data showed that the inflamed endometrium presented low CYP19A1 and GPER expression, and significantly higher EMT process versus the normal tissue. Moreover, we established a TGF-ß1-induced fibrosis model in BEND cells, and found that E2 inhibited the EMT process of BEND cells in a dose-dependent manner. The anti-fibrotic effect of E2 was blocked by the GPER inhibitor G15, but not the estrogen nuclear receptors (ERs) inhibitor ICI182780. Moreover, the GPER agonist G1 inhibited fibrosis and Smad2/3 phosphorylation but increased the expression of TGFBR3 in BEND cells. Transfection with TGFBR3 small interfering RNA blocked the effect of G1 on fibrosis of BEND cells and upregulated the expression of P-Smad2/3. Our in vivo data also showed that E2 and G1 affected uterus fibrosis in mice endometritis model caused by LPS, which was associated with the inhibition of TGFBR3/Smad2/3 signaling. In conclusion, our data implied that E2 alleviates the fibrosis of TGF-ß1-induced BEND cells, which is associated with the GPER mediation of TGFBR3/Smad2/3 signaling.


Asunto(s)
Endometritis , Estradiol , Proteoglicanos , Receptores de Factores de Crecimiento Transformadores beta , Factor de Crecimiento Transformador beta1 , Animales , Bovinos , Femenino , Ratones , Endometritis/metabolismo , Endometrio/metabolismo , Células Epiteliales/metabolismo , Transición Epitelial-Mesenquimal , Estradiol/farmacología , Estrógenos/metabolismo , Fibrosis , Receptores Acoplados a Proteínas G/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Proteínas Smad/metabolismo
2.
J Reprod Dev ; 68(1): 21-29, 2022 Feb 18.
Artículo en Inglés | MEDLINE | ID: mdl-34690214

RESUMEN

Inadequate fetomaternal interactions could directly lead to pregnancy failure in dairy cows. Exosomes are widely involved in endometrial matrix remodeling, immune function changes, placental development, and other processes of embryo implantation and pregnancy in dairy cows. However, the role of exosomes derived from placental trophoblast cells in regulating the receptivity of endometrial cells and facilitating fetomaternal interaction remains unclear. In this study, bovine trophoblast cells (BTCs) were obtained from bovine placenta and immortalized by transfection with telomerase reverse transcriptase (TERT). Immortalized BTCs still possess the basic and key properties of primary BTCs without exhibiting any neoplastic transformation signs. Subsequently, the effect of trophoblast-derived exosomes (TDEs) on endometrial receptivity in endometrial epithelial cells (EECs) was determined, and the mechanism whereby TDEs and their proteins participate in the fetomaternal interaction during bovine pregnancy were explored. EECs were co-cultured with the exosomes derived from BTCs treated with progesterone (P4). Such treatment enhanced the expression of the endometrial receptivity factors, integrin αv, ß3, Wnt7a, and MUC1 by changing the extracellular environment, metabolism, and redox balance in EECs via proteome alignment, compared with no treatment according to the DIA quantitation analysis. Our study demonstrated that trophoblast-derived exosome proteins are one of the most critical elements in fetomaternal interaction, and their changes may act as a key signal in altering endometrial receptivity and provide a potential target for improving fertility.


Asunto(s)
Exosomas , Trofoblastos , Animales , Bovinos , Implantación del Embrión/fisiología , Endometrio/metabolismo , Células Epiteliales/metabolismo , Exosomas/metabolismo , Femenino , Placenta , Embarazo , Trofoblastos/metabolismo
3.
Reprod Domest Anim ; 57(11): 1406-1417, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-35881670

RESUMEN

During maturation, spermatozoa acquire motility and fertilizing capacity as they transit through the epididymis. Melatonin is a lipophilic hormone with multiple functions in regulating the fertility. Previous studies have shown that melatonin affected the capacitation or maturation of sperm in the epididymis. The aim of this study was to investigate the effects of melatonin on epididymal caput epithelial cells in sheep. In the study, we used iTRAQ labelling coupled with LC-MS/MS for quantitative identification of differentially expressed proteins in melatonin-treated sheep epididymal caput epithelial cells. We identified 69 differentially expressed protein; 41 were upregulated and 28 were downregulated in samples from sheep in melatonin treated. We validated the differential expression of a subset of these proteins using qPCR and Western blot. Gene ontology annotation identified that the differentially expressed proteins function in cellular processes and metabolic processes. Notably, five of the differentially expressed proteins as SOD1, COL1A1, PRM1, NQO2, and FN1 are involved in sperm migration and sperm maturation. KEGG enrichment analysis demonstrated significant enrichment in several cardiac-related pathways, such as "PI3K-Akt signaling pathway", "AGE-RAGE signaling pathway in diabetic complications", "ECM-receptor interaction", and "Ribosome". Our results suggest that candidate biomarker (SOD1, COL1A1, PRM1, NQO2, and FN1) discovery can aid in understanding sperm development and maturation in sheep. These results provide insights into the potential mechanisms of melatonin regulation of sperm maturation in epididymal caput epithelial cells.


Asunto(s)
Epidídimo , Melatonina , Masculino , Ovinos , Animales , Epidídimo/metabolismo , Melatonina/farmacología , Melatonina/metabolismo , Proteómica , Cromatografía Liquida/veterinaria , Fosfatidilinositol 3-Quinasas/metabolismo , Superóxido Dismutasa-1/metabolismo , Semen , Espectrometría de Masas en Tándem/veterinaria , Maduración del Esperma/fisiología , Espermatozoides/fisiología , Proteínas/metabolismo , Células Epiteliales
4.
Reprod Domest Anim ; 57(9): 1046-1055, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35678492

RESUMEN

Cryptorchidism, as a common congenital disease of canine testes, is mainly caused by factors leading to endocrine abnormalities in testes and infertility in a heat stress and hypoxia microenvironment. Moreover, heat stress and hypoxia, as critical microenvironmental factors, promote epithelial-mesenchymal transition (EMT), which occurs during adult tissue remodelling responses including carcinogenesis and fibrosis and is the main cause of testicular tumours. In this study, we found by haematoxylin-eosin staining that the canine cryptorchid tissue produced a lot of collagen fibres. Also, the quantitative PCR and Western blot results showed that the mRNA and protein levels of the heat stress makers HSP70 and HO-1 and the hypoxia maker HIF-1α are significant higher compared with normal testes. Moreover, we found the expression levels of TGF-ßs and its two receptors TGF-ßRI and TGF-ßRII increased in case of cryptorchidism. From the study in vitro, we found both heat stress and COCl2 mimic hypoxia inhibited the secretion of testosterone (T) and androstenedione (A4) and promoted the expression of the EMT maker α-SMA and vimentin in Leydig cells, and also that heat stress and COCl2 stimulated with the TGF-ß signalling promoted the expression of TGF-ßs and its two type receptors and also the active phosphorylation of Smad2 and Smad3. The use of LY2109761, a receptor inhibitor of TGF-ßs/Smad signalling pathway, was associated with heat stress and COCl2 suppression of androgens' secretion and stimulated EMT in Leydig cells. These findings characterized a novel pathogenesis of cryptorchidism and provided a new idea for therapeutics.


Asunto(s)
Criptorquidismo , Enfermedades de los Perros , Andrógenos , Animales , Criptorquidismo/veterinaria , Perros , Transición Epitelial-Mesenquimal , Respuesta al Choque Térmico , Hipoxia/veterinaria , Masculino , Factor de Crecimiento Transformador beta , Factor de Crecimiento Transformador beta1/farmacología
5.
Sensors (Basel) ; 22(1)2022 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-35009894

RESUMEN

For the optimization of three-phase hybrid stepper motors with complex electromagnetic structures, an optimization method is presented in this paper. The method is a combination of 3D-FEM and the Taguchi optimization method intended to reduce the dependence on FEM results during the optimization calculation. In this paper, the optimization method is used in the optimization of the tooth shape of the three-phase hybrid stepper motor, and the objective is to reduce the noise caused by harmonics in the "torque-angle characteristic" of the motor. It is clear that traditional optimization methods make it very difficult to carry out such an optimization calculation as a large number of finite element calculations have to be used in the optimization process, and the required computation time is extremely long. Using the optimization method presented in the paper, the optimization becomes feasible because the number of finite element calculations is greatly reduced and the computation time is thus greatly reduced. In order to check the effectiveness of the optimization, the waterfall diagram for noise analysis and its application to check torque ripple are also presented in the paper. Both simulation and test results show that the optimized structure can significantly reduce the motor noise caused by torque ripple. Therefore, the optimization method proposed in this paper can be an effective tool for the optimal design of high-performance motors, including stepper motors.

6.
J Cell Physiol ; 236(6): 4565-4579, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33230823

RESUMEN

Endometritis is a common disease affecting fertility in cows during the perinatal period, which disturbs the molecular milieu of the uterine environment and impairs embryo development and implantation. Exosomes are important extracellular components that transmit a variety of micro RNAs (miRNAs), which perform key regulatory functions. In this study, we investigated plasma exosomal miRNAs from cows with endometritis and from cultured endometrial epithelial cells (EECs) challenged with lipopolysaccharide (LPS) to explore the role of EEC-derived exosomes and their miRNAs in bovine endometritis. Plasma exosomes were collected from nine healthy dairy cows and nine dairy cows with endometritis, and culture supernatant exosomes were isolated from EECs challenged with or without LPS. Exosomal RNA was extracted using commercial kits and miRNA profiles were generated using RNA-seq. We found that miR-218 was differentially expressed in EECs under conditions of endometrial inflammation. Inhibition studies suggested that reduced levels of miR-218 in EEC-derived exosomes when transferred into placental trophoblast cells impaired embryonic development and decreased placental trophoblast cell migration by targeting secreted frizzled related protein 2. We propose that exosomal miR-218 secreted from EECs acts as a driver of embryonic development and differentiation. In addition, exosomal miR-218 may provide a valuable diagnostic marker for bovine endometritis.


Asunto(s)
Endometritis/metabolismo , Endometrio/metabolismo , Células Epiteliales/metabolismo , Exosomas/metabolismo , Proteínas de la Membrana/metabolismo , MicroARNs/metabolismo , Trofoblastos/metabolismo , Animales , Apoptosis , Bovinos , Movimiento Celular , Células Cultivadas , Endometritis/genética , Endometritis/patología , Endometrio/efectos de los fármacos , Endometrio/patología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Exosomas/efectos de los fármacos , Exosomas/genética , Exosomas/patología , Femenino , Fertilización In Vitro , Regulación del Desarrollo de la Expresión Génica , Técnicas de Maduración In Vitro de los Oocitos , Lipopolisacáridos/farmacología , Proteínas de la Membrana/genética , MicroARNs/genética , Embarazo , Transducción de Señal
7.
J Cell Physiol ; 236(10): 6793-6805, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-33748999

RESUMEN

Skeletal muscle development is a complex biological process involving multiple key genes, signaling pathways and noncoding RNAs, including microRNAs and circular RNAs (circRNAs). However, the regulatory relationship among them is so complicated that it has not yet been fully elucidated. In this study, we found that miR-7 inhibited C2C12 cell proliferation and differentiation by targeting transcription factor 12 (TCF12). circHIPK3 acted as a competing endogenous RNA, and its overexpression effectively reversed the regulation of miR-7 on C2C12 cell proliferation and differentiation by increasing TCF12 expression. Taken together, our findings provide evidence that circHIPK3 regulates skeletal muscle development through the miR-7/TCF12 pathway. This study provides a scientific basis for further research on skeletal muscle development at the circRNA level.


Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Diferenciación Celular , Proliferación Celular , MicroARNs/metabolismo , Desarrollo de Músculos , Mioblastos Esqueléticos/metabolismo , ARN Circular/metabolismo , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Línea Celular , Regulación de la Expresión Génica , Ratones , MicroARNs/genética , ARN Circular/genética , Transducción de Señal
8.
Appl Opt ; 60(11): 3182-3186, 2021 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-33983217

RESUMEN

Semi-insulating (SI) SiC photoconductive semiconductor switches were prepared using two compensation mechanisms: namely vanadium dopants compensation (4H- and 6H-SiC) and deep level defect compensation (4H-SiC). The bias voltage and current of the high-purity (HP) SI 4H-SiC photoconductive semiconductor switch (PCSS) with a channel length of 1 mm reached 24 kV and 364 A, respectively, and the minimum on-state resistance of approximately 1 Ω was triggered by laser illumination at a wavelength of 355 nm. The experimental results show that, in this case, the on-state characteristics of HP 4H-SiC PCSS are superior to those of the vanadium-doped(VD) 4H and 6H-SiC PCSS devices. HP 4H-SiC PCSS shows remarkable waveform consistency. Unlike for VD 4H and 6H-SiC PCSS, the current waveform of HP 4H-SiC PCSS exhibits a tailing phenomenon due to its longer carrier lifetime.

9.
Reprod Domest Anim ; 56(2): 292-300, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33001490

RESUMEN

Steroid hormones and receptors play important roles in female reproduction, and their expression patterns affect follicular growth and development. To examine the expression of dihydrotestosterone (DHT) synthases (5α-reductases (5α-red1 and 5α-red2)) and androgen receptor (AR) during follicular development, and the regulation of DHT signalling by follicle-stimulating hormone (FSH) and luteinizing hormone (LH), we have used enzyme-linked immunosorbent assays, quantitative real-time polymerase chain reaction, immunohistochemical staining and Western blotting to examine DHT synthesis in small (≤2 mm), medium (2-5 mm) and large (≥5 mm) sheep follicles. Expression of 5α-red1, 5α-red2 and AR was observed in ovine ovaries, and with the development of follicles, the expressions of 5α-red1 and 5α-red2 mRNA and protein increased, but the levels of AR mRNA, protein and DHT level decreased. In addition, granulosa cells were treated with FSH (0.01, 0.1 and 1 international unit (IU)/ml), LH (0.01, 0.1 and 1 IU/ml) and testosterone (T, 10-7  M) to evaluate the effects of FSH and LH on DHT and oestradiol (E2) synthesis and 5α-red1, 5α-red2 and AR expression. We found that FSH and LH upregulated 5α-red1 and 5α-red2 in sheep granulosa cells, but downregulated the concentration of DHT and expression of AR. Meanwhile, FSH and LH significantly upregulated the expression of aromatase (P450arom) and secretion of E2. This result indicates that although FSH and LH promote the expression of 5α-red1 and 5α-red2, T is not transformed into DHT, but E2. This study reveals the reason why DHT concentration is downregulated in large follicles and lays a foundation for further exploring the synthesis mechanism of DHT during follicular development.


Asunto(s)
Dihidrotestosterona/metabolismo , Hormona Folículo Estimulante/farmacología , Células de la Granulosa/metabolismo , Hormona Luteinizante/farmacología , Animales , Femenino , Folículo Ovárico/metabolismo , Oxidorreductasas/metabolismo , Receptores Androgénicos/metabolismo , Oveja Doméstica
10.
Exp Cell Res ; 385(1): 111669, 2019 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-31605696

RESUMEN

PURPOSE: The main objective of our study was to explore changes in the expression levels of differentially expressed genes associated with prostate cancer progression and to design a series of experiments to verify the function of differentially expressed genes. METHOD: The transcriptome datas of 499 cases of prostate cancer patients was downloaded from TCGA database. Differential genes associated with Gleason score were selected and filtered out by p < 0.05 and spearman coefficient >0.3. KEGG signaling pathway was enriched by differentially expressed genes, and TTK was selected as the research object. The expression of TTK was tested in prostate cancer tissues and prostate cancer cell lines. The changes of biological behavior of prostate cancer cell lines were verified after TTK was knocked out by siRNA and tumorigenic effect of TTK was verified by shRNA in vivo experiments. RESULT: The expression of TTK was positively correlated with Gleason score of prostate cancer, and the expression of protein and mRNA in metastatic prostate cancer cell lines was higher than that in non-metastatic prostate cancer cell lines. Vitro biological experiments showed that TTK gene knockout could inhibit the proliferation, invasion and migration of PC3 and DU145 cells, and promote cell apoptosis. In vivo experiments showed that TTK knockout inhibited tumorigenesis in mice. It was found that the expression of CDK2 and CCNE1 decreased after TTK was knocked out. CONCLUSION: Our results suggest that TTK is a gene associated with malignancy of PCa and could be a novel therapeutic target for clinical application.


Asunto(s)
Proteínas de Ciclo Celular/genética , Proliferación Celular/genética , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Tirosina Quinasas/genética , Animales , Apoptosis/genética , Carcinogénesis/genética , Carcinogénesis/patología , Línea Celular Tumoral , Quinasa 2 Dependiente de la Ciclina/genética , Progresión de la Enfermedad , Humanos , Masculino , Ratones , Clasificación del Tumor , Proteínas Oncogénicas/genética , Células PC-3 , Próstata/patología , ARN Interferente Pequeño/genética , Transducción de Señal/genética , Transcriptoma/genética
11.
Gen Comp Endocrinol ; 286: 113245, 2020 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-31415730

RESUMEN

Dihydrotestosterone (DHT) and 17ß-estradiol (E2) are sex hormones that regulate human hair follicle (HF) growth and are produced by peripheral reduction and aromatization of testosterone. However, the expression patterns of DHT and E2 synthesis-related proteins and their receptors in male yak skin during different HF stages (telogen, anagen, and catagen) are unknown. In this study, we found that both 5α-red and androgen receptor (AR) were expressed in epithelial cells and AR was expressed in the dermal papilla. Additionally, the transcription level of 5α-red1 at different HF stages was significantly higher than that of 5α-red2 mRNA at the same stage; 5α-red1 and 5α-red2 proteins peaked during the anagen and telogen periods of HF, respectively. However, AR protein was only expressed in the skin during the anagen phase of HF. Aromatase and estrogen receptors (ERα and ERß) were expressed in cutaneous epithelial cells, whereas ERα and ERß were expressed in the dermal papilla; the transcription level of ERα in HFs at each stage was much higher than that of ERß. From the catagen to telogen phase, aromatase protein expression was down-regulated, while ERα protein expression was up-regulated. Based on our results, we speculate that 5α-red1 is essential for the synthesis of DHT in male yak skin epithelial cells and promotes the growth of HFs through AR. E2 synthesized by male yak skin epithelial cells may inhibit the growth of male yak skin HFs by ERα. These results provide a foundation for further study on the mechanism of hormone-regulated male yak skin HFs.


Asunto(s)
Antígeno 12E7/metabolismo , Dihidrotestosterona/metabolismo , Folículo Piloso/crecimiento & desarrollo , Animales , Bovinos , Folículo Piloso/metabolismo , Masculino
12.
Gen Comp Endocrinol ; 286: 113135, 2020 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-30849411

RESUMEN

Some of the functions of melatonin in mammals are exerted through its membrane receptors (MRs) and studies have shown that estradiol (E2) might play an important role in regulating the expression of these proteins in female reproductive organs. However, no reports have reported the expression of MRs in the sheep oviduct or whether they are regulated by E2. Thus, herein, we detected the localization of MT1 and MT2 in the sheep oviduct. Moreover, we also investigated the expression pattern of these markers in the ovulating and non-ovulating side of the oviduct in the sheep ampulla and isthmus. Immunohistochemistry analyses revealed that both MT1 and MT2 are mainly expressed on oviduct epithelial cells. Both real-time polymerase chain reaction (qPCR) and western blot analyses showed that MT1 and MT2 genes and proteins are highly expressed on the non-ovulating side of the oviduct ampulla, but not the ovulating side. However, regarding the oviduct isthmus, there were no significant differences between the ovulating and non-ovulating sides. In vitro, 10 ng/ml and 1 µg/ml of E2, as well as 1 µg/ml of E2 combined with 0.1 µg/ml, 1 µg/ml, and 10 µg/ml of ICI182780 (a non-selective estrogenreceptor antagonist), were used to treat oviduct epithelial cells. We found that E2 inhibited the expression of MT1 and MT2 in cultured oviduct cells. Moreover, the inhibitory effect was suppressed by ICI182780. In conclusion, it was demonstrated that MRs are present in the sheep oviduct, and that E2, via the ER pathway, regulates their expression in the oviduct.


Asunto(s)
Oviductos/metabolismo , Receptores de Melatonina/metabolismo , Animales , Femenino , Humanos , Ovinos
13.
Reprod Domest Anim ; 55(4): 460-468, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-31958357

RESUMEN

The androgen receptor (AR) plays a key role in reproduction, and aromatase (P450arom), nuclear oestrogen receptors (ERs) α and ß, and G protein-coupled receptor 30 (GPR30) are important for testicular and epididymal cell proliferation and development. In the study, we have investigated the expression and localization of AR, P450arom, ERα, ERß and GPR30 in testes and epididymides of sexually mature sheep by quantitative reverse transcription-polymerase chain reaction, Western blotting and immunohistochemistry. The results demonstrate that the AR, P450arom and ERα levels in the caput and corpus epididymis were significantly lower than those in the testis and cauda epididymis (p < .05), the ERß level in the testis was significantly higher than in the caput, corpus and cauda epididymis (p < .05), and the GPR30 level in the caput epididymis was significantly lower than in the testis and corpus and cauda epididymis (p < .05). These receptors were mainly detected in epididymal epithelial, basal, smooth muscle, Sertoli and Leydig cells, as well as in spermatozoa. Taken together, the results suggest that sheep epididymides and testes have the potential for estradiol synthesis and are the targets of both androgens and estradiol. These results provide a foundation for further studies on the mechanisms of androgens and estradiol signalling in the testes and epididymides of sheep.


Asunto(s)
Epidídimo/metabolismo , Ovinos , Testículo/metabolismo , Animales , Aromatasa/metabolismo , Receptor alfa de Estrógeno/metabolismo , Receptor beta de Estrógeno/metabolismo , Masculino , Receptores Androgénicos/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Espermatozoides/metabolismo , Distribución Tisular
14.
J Cell Physiol ; 234(9): 15182-15193, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-30671954

RESUMEN

Corpus luteum (CL) regression is a complex physiological process. Previous studies have shown that dihydrotestosterone (DHT) may be involved in regulating CL regression, but the mechanism is still unclear. In this study, we evaluated the localization of the two isoforms of DHT synthetase 5α-reductase (5α-red1 and 5α-red2) and androgen receptor (AR) in sheep CL, and investigated 5α-red1, 5α-red2, AR, and DHT levels at different luteal stages of CL (early, middle, and late phase) by immunohistochemistry, quantitative real-time polymerase chain reaction, and western blot analysis. Moreover, we cultured luteal cells from middle phase CL and treated them with different concentrations of DHT (10-10 -10 -6 M) and the AR antagonist flutamide (10 -5 M), to evaluate whether DHT is involved in the regulation of progesterone (P4) secretion and progesterone nuclear receptor (PGR) expression and whether these effects are regulated by the AR pathway. We also investigated the effects of DHT and flutamide on prostaglandin F2α (PGF2α) secretion and apoptotic gene and protein expression. Our results showed that 5α-red1, 5α-red2, and AR were expressed in the CL, and their expression and DHT levels were changed during the luteal phase. DHT was involved in mediating P4 and PGF2α secretion and PGR and apoptotic gene and protein expression. The effects of DHT on CL were at least partially regulated by the AR pathway. This study reveals the mechanism of action of DHT on sheep CL regression and lays the foundation for further exploration of androgen regulation of CL function.

15.
Microb Pathog ; 130: 137-145, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30858008

RESUMEN

The silkworm (Bombyx mori) is a typical and economically important lepidopteran species, and research has resulted in the development and accumulation of breeding lines. Studies of immune-related silkworm genes not only promote our understanding of silkworm immune response mechanisms, but they also inform insect immune molecular diversity research. Here, silkworm proteins were screened using proteomics after Bombyx mori nuclear polyhedrosis virus (BmNPV) infection, and 2368 silkworm proteins were identified, including six antimicrobial peptides and 12 serpins. The mRNA expression levels of these 18 proteins were examined at different times. The results indicated that attacin had the highest expression level, while serpin-5 and cecropin-D exhibited a negative regulatory correlation. These results provide a significant step toward a deeper understanding of B. mori immunoregulation.


Asunto(s)
Péptidos Catiónicos Antimicrobianos/análisis , Bombyx/inmunología , Bombyx/virología , Proteínas de Insectos/análisis , Nucleopoliedrovirus/crecimiento & desarrollo , Serpinas/análisis , Animales , Perfilación de la Expresión Génica , Proteoma/análisis , ARN Mensajero/análisis
16.
Reprod Biol Endocrinol ; 17(1): 1, 2019 Jan 03.
Artículo en Inglés | MEDLINE | ID: mdl-30606208

RESUMEN

BACKGROUND: Melatonin is an amine hormone that plays an important role in regulating mammalian reproduction. This study aimed to investigate the expression pattern of melatonin synthesis enzymes AANAT and HIOMT and melatonin receptors MT1 and MT2 in sheep cumulus-oocyte complexes (COCs) as well as the change of melatonin level in follicular fluid (FF) during antral follicle development. In this research, we also study the effect of ß-estradiol (E2) on MT1 and MT2 expression as well as melatonin synthesis in COCs so as to lay the foundation for further exploration of the regulation mechanism of melatonin synthesis in the ovary. METHODS: COCs and FF were collected from different size (large follicles (diameter ≥ 5 mm), medium follicles (diameter 2-5 mm), and small follicles (diameter ≤ 2 mm)) of antral follicles in sheep ovaries. To assess whether E2 regulates melatonin synthase and its receptors expression in sheep COCs and whether it is mediated through estrogen receptor (ER) pathway. The collected COCs were cultured in vitro for 24 h and then treat with 1 µM E2 and/or 1 µM ICI182780 (non-selective ER antagonist). The expression of AANAT, HIOMT, MT1 and MT2 mRNA and protein were determined by qRT-PCR and western blot. The melatonin level was determined by ELISA. RESULTS: The expression of AANAT, HIOMT, MT1 and MT2 were significantly higher expression in the COCs of small follicles than in those of large follicles (P < 0.05). However, the melatonin level was significantly higher in large follicle FF than in small follicle FF (P < 0.05). Further, the expression of AANAT, HIOMT, MT1, and MT2 and melatonin production were decreased by E2 treatment (P < 0.05), but when ICI182780 was added, the expression of AANAT, HIOMT, MT1, and MT2 and melatonin production recovered (P < 0.05). CONCLUSIONS: We suggest that sheep COCs can synthesize melatonin, but this ability is decreased with increasing follicle diameter. Furthermore, E2 play an important role in regulated the expression of MT1 and MT2 as well as melatonin synthesis in sheep COCs through the ER pathway.


Asunto(s)
Células del Cúmulo/metabolismo , Estradiol/farmacología , Melatonina/biosíntesis , Oocitos/metabolismo , Receptores de Melatonina/metabolismo , Ovinos/metabolismo , Animales , Vías Biosintéticas/efectos de los fármacos , Estradiol/metabolismo , Estradiol/fisiología , Femenino , Líquido Folicular/metabolismo , Melatonina/metabolismo , Folículo Ovárico/metabolismo , Receptores de Estrógenos/metabolismo
17.
Appl Opt ; 58(31): 8465-8470, 2019 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-31873330

RESUMEN

Fiber-optic Fabry-Perot pressure sensors based on silicon diaphragms of different thicknesses were fabricated using surface and bulk MEMS techniques in this study. The multi-beam interference resulting from multiple reflecting mirrors with the elastic deformation of the Fabry-Perot sensor was simulated by finite element analysis. The pressure sensitivities of the sensors with different diaphragm thicknesses and the relationship between the pressure and the wavelength shift were simulated. The simulation results were in good agreement with the test results. This study provides guidance for future sensor models and parameter design.

18.
Reprod Domest Anim ; 54(10): 1305-1312, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31188500

RESUMEN

Oestrogen, androgen and progesterone are involved in the regulation of uterine physiological functions, with the participation of the following proteins: oestrogen receptor (ER), androgen receptor (AR) and progesterone nuclear receptor (PGR). In this study, we used immunohistochemistry to detect the localization of ERα, ERß, AR and PGR in sheep uterus. Additionally, we used real-time polymerase chain reaction (RT-qPCR) and Western blot technique to analyse their expression profiles at different stages of sheep oestrous cycle in the endometrium and myometrium. Immunohistochemical analysis showed that ERα, ERß, AR and PGR were present in sheep uterus in oestrus, mainly in the uterine luminal epithelium, stroma, gland and myometrium. Real-time polymerase chain reaction results showed that in the endometrium, ERα expression level was highest in oestrus. ERß and PGR, instead, were highly expressed in pro-oestrus. In the myometrium, ERα was highly expressed in both oestrus and pro-oestrus, and ERß was highly expressed in oestrus and dioestrus. Progesterone nuclear receptor expression was highest in oestrus, followed by metoestrus. In the endometrium, both receptors ERα and ERß were abundant in pro-oestrus, while the maximum AR protein content was found in oestrus. At this stage of the oestrous cycle, PGR protein concentration in the myometrium was significantly lower than those observed in other stages. These results suggest that these receptors are important for sheep reproductive function, as their expression at mRNA and protein levels exhibits particular time- and tissue-specific profiles along the oestrous cycle.


Asunto(s)
Receptores Androgénicos/metabolismo , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/metabolismo , Ovinos , Útero/metabolismo , Animales , Ciclo Estral/genética , Ciclo Estral/metabolismo , Femenino , Perfilación de la Expresión Génica , Receptores Androgénicos/genética , Receptores de Estrógenos/genética , Receptores de Progesterona/genética
19.
Sensors (Basel) ; 19(21)2019 Nov 03.
Artículo en Inglés | MEDLINE | ID: mdl-31684134

RESUMEN

Silicon-diaphragm-based fiber-optic Fabry-Perot sensors with different intracavity pressures were fabricated by anodic bonding and microelectromechanical techniques. The thermal stress and thermal expansion of the Fabry-Perot (FP) sensor caused by high-temperature bonding and temperature change were simulated by finite-element analysis. The calculated thermal stress is largest in the center and edge regions of the resonance cavity, reaching from 2 to 6 MPa. The reflection spectra and temperature sensitivity of the sensors were simulated by using a two-dimensional wave-optic model in Comsol. Theoretical calculations were also made for the FP cavity without considering silicon-diaphragm deformation and thermal stress. Four sensors with intracavity pressures of 0.01, 0.03, 0.04, and 0.05 MPa were tested at low temperatures, showing a high degree of consistency with the simulation results rather than theoretical calculation, especially for high intracavity pressure. This method is expected to aid the analysis of thermal stress generated during the bonding process and to facilitate better design and control of the temperature sensitivity of the sensor.

20.
Appl Opt ; 57(11): 2804-2808, 2018 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-29714282

RESUMEN

High-power photoconductive semiconductor switching devices were fabricated from a high-purity, semi-insulating 4H-SiC wafer. A highly n-doped GaN subcontact layer was inserted between the contact metal and the high-resistivity SiC wafer. The minimum ON-state resistance of the device was less than 1 ohm when the energy of a 355 nm laser was 10.5 mJ with a bias voltage of 6 kV. The maximum device lifetime is 3151 pulses, after which the device completely fails. The failure mechanisms are determined using several analysis methods. Under a strong electric field, the failure mechanism differs for the two electrodes. Near the edge of the anode electrode, the switch is damaged due to the thermal stress caused by impact ionization. At the edge of the cathode electrode, the electrode erosion is the main reason for the failure to operate for long periods of time. These two different damage mechanisms are both important factors influencing the device performance. The electron avalanche breakdown at the edge of the anode electrode causes the formation of cracks between the electrodes, which is the root cause of the switch failure.

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