RESUMEN
Objective: We aim to evaluate the morbidity and mortality of cancer attributable to human papillomavirus (HPV) infection in China in 2016. Methods: Based on the cancer incidence and mortality rates, national population data, and population attributable fraction (PAF) in China, we calculated the number of incidence and death cases attributed to HPV infection in different areas, age groups, and gender in China in 2016. The standardized incidence and mortality rates for cancer attributed to HPV infection were calculated by using Segi's population. Results: In 2016, a total of 124 772 new cancer cases (6.32 per 100 000) were attributed to HPV infection in China, including 117 118 cases in women and 7 654 cases in men. Of these cancers, cervical cancer was the most common one, followed by anal cancer, oropharyngeal cancer, penile cancer, vaginal cancer, laryngeal cancer, oral cancer, and vulvar cancer. A total of 41 282 (2.03 per 100 000) deaths were attributed to HPV infection, of which 37 417 occurred in women and 3 865 in men. Most deaths were caused by cervical cancer, followed by anal cancer, oropharyngeal cancer, penile cancer, laryngeal cancer, vaginal cancer, oral cancer, and vulvar cancer. The incidence and mortality rates of cervical cancer increased rapidly with age, peaked in age group 50-54 years, then decreased obviously. The morbidity and mortality rates of non-cervical cancer increased with age. The cancer case and death numbers in rural areas (57 089 cases and 19 485 deaths) were lower than those in urban areas (67 683 cases and 21 797 deaths). However, the age-standardized incidence rate (ASIR) and age-standardized mortality rate (ASMR) of cervical cancer were higher in rural areas than in urban areas. There were no significant differences in ASIR and ASMR of non-cervical cancers between urban areas and rural areas. Conclusions: The incidence of cancers attributed to HPV infection in China was lower than the global average, but the number of incidences accounted largely, furthermore there is an increasing trend of morbidity and mortality. The preventions and controls of cervical cancer and male anal cancer are essential to contain the increases in cancer cases and deaths attributed to HPV infection.
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Neoplasias Laríngeas , Neoplasias de la Boca , Neoplasias Orofaríngeas , Infecciones por Papillomavirus , Neoplasias del Pene , Neoplasias del Cuello Uterino , Neoplasias Vaginales , Neoplasias de la Vulva , China/epidemiología , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Neoplasias Orofaríngeas/epidemiología , Infecciones por Papillomavirus/epidemiología , Neoplasias del Pene/epidemiología , Sistema de Registros , Neoplasias del Cuello Uterino/epidemiologíaRESUMEN
Objective: To explore the effect of nutritional status pre-and during chemoradiotherapy on the prognosis of patients with limited- stage small cell lung cancer (LS-SCLC). Methods: We retrospectively collected medical records of 172 LS-SCLC patients undergoing concurrent chemoradiotherapy in our hospital from 2000 to 2014, with 126 males and 46 females. The data of complete blood count and hepatic and renal function were collected before initial treatment, before radiotherapy, 4 weeks during radiotherapy, and 1 month after complete of treatment. The prognostic nutritional index(PNI)was calculated. Kaplan-Meier method was used to calculate the survival rate. Log-rank test was performed used to compare the survival differences between groups. Multivariate prognostic analysis was performed using Cox regression model. Results: The median overall survival (OS) was 21 months, with median progression-free survival (PFS) of 11 months. At the beginning of treatment, patients with pre-treatment PNI ≥ 53 had significantly superior OS (median 37 vs 15 months, P=0.001) and PFS (median 16 vs 10 months, P=0.017). Patients with pre-treatment hemoglobin ≥140 g/L and <140 g/L had an median OS of 32 months and 17 months (P=0.019), and median PFS of 16 months and 9 months (P=0.040), respectively. During chemoradiation, patients with elevated hemoglobin had similar median OS compared with those had decreased hemoglobin (27 vs 18 months, P=0.063, but superior median PFS (15 vs 9 months, P=0.017). Multivariate analysis revealed that prophylactic cranial irradiation, pre-treatment hemoglobin ≥140 g/L, and pretreatment PNI ≥53 were independent predictors of OS and PFS in patients with LS-SCLC. Conclusion: Pre-treatment nutritional status and the changes of nutritional status during chemoradiotherapy is significantly associated with the prognosis of patients with limited-stage small cell lung cancer. The patients with better pre-treatment nutritional status have a better prognosis.
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Carcinoma de Células Pequeñas/tratamiento farmacológico , Carcinoma de Células Pequeñas/radioterapia , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/radioterapia , Estado Nutricional , Carcinoma Pulmonar de Células Pequeñas/patología , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma de Células Pequeñas/química , Quimioradioterapia , Terapia Combinada , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , Estudios Retrospectivos , Carcinoma Pulmonar de Células Pequeñas/mortalidad , Tasa de SupervivenciaRESUMEN
A new flavonol, tonkinensisol, was isolated from the roots of Sophora tonkinensis, together with three known compounds named as bayin, vitexin and lupeol. Their structures were elucidated on the basis of spectroscopic evidence. Additionally, tonkinensisol showed moderate cytotoxicity suppressing the proliferation of HL-60 cells in vitro.
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Antineoplásicos Fitogénicos/aislamiento & purificación , Flavonoles/aislamiento & purificación , Sophora/química , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Proliferación Celular/efectos de los fármacos , China , Flavonoles/química , Flavonoles/farmacología , Células HL-60 , Humanos , Espectrometría de Masas , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Raíces de Plantas/química , Espectrofotometría Infrarroja , Espectrofotometría UltravioletaRESUMEN
We previously reported an increased secretion of amyloid precursor-like protein 2 (APLP2) in the healing corneal epithelium. The present study sought to investigate signal transduction pathways involved in APLP2 shedding in vitro. APLP2 was constitutively shed and released into culture medium in SV40-immortalized human corneal epithelial cells as assessed by Western blotting, flow cytometry, and indirect immunofluorescence. Activation of protein kinase C (PKC) by phorbol 12-myristate 13-acetate (PMA) caused significant increases in APLP2 shedding. This was inhibited by staurosporine and a PKC-epsilon-specific, N-myristoylated peptide inhibitor. Epidermal growth factor (EGF) also induced APLP2 accumulation in culture medium. Basal APLP2 shedding as well as that induced by PMA and EGF was blocked by a mitogen-activated protein kinase (MAPK) kinase inhibitor, U-0126. Our results suggest that MAPK activity accounts for basal as well as PKC- and EGF-induced APLP2 shedding. In addition, PKC-epsilon may be involved in the induction of APLP2 shedding in corneal epithelial cells.
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Precursor de Proteína beta-Amiloide/metabolismo , Epitelio Corneal/metabolismo , Proteínas Quinasas Activadas por Mitógenos/fisiología , Proteínas del Tejido Nervioso/metabolismo , Línea Celular Transformada , Factor de Crecimiento Epidérmico/farmacología , Epitelio Corneal/citología , Epitelio Corneal/efectos de los fármacos , Humanos , Quinasas de Proteína Quinasa Activadas por Mitógenos/fisiología , Fosforilación , Proteína Quinasa C/fisiología , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
The main goal of the present study was to investigate the response of cultured bovine corneas to the application of irritant substances and its potential use for predicting ocular irritancy in humans. We hypothesized that chemicals causing eye irritation may induce disruption of epithelial tight junctions and trigger cell stress responses modulated via transcription factors such as AP-1 and NF-kappaB. A simple air-lifted corneal organ culture system was used as an ex vivo model for ocular irritancy test. The effects of two surfactants, sodium dodecyl sulfate (SDS) and benzalkonium chloride (BAK), on corneal epithelial permeability and DNA-binding activity of AP-1 and NF-kappaB were studied in cultured bovine corneas. Both SDS and BAK induced tight junction disruption and increased permeability of corneal epithelium assessed using surface biotinylation in a concentration- and time-dependent manner. An increase in DNA-binding activity measured using electrophoretic mobility shift assay was observed when cultured corneas were treated with surfactants at concentrations causing minimal to mild ocular irritation, indicating epithelial cell stress response. Furthermore, exposure of cultured corneas to SDS or BAK at concentrations causing severe ocular irritancy resulted in a decrease in DNA-binding activity of these transcription factors in epithelial cells. These results indicate that the combination of corneal organ culture and measurements of corneal epithelial permeability and DNA-binding activity of stress-response transcription factors following chemical exposure has the potential to be used as a mechanistically based alternative to in vivo animal testing.
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Córnea/efectos de los fármacos , Irritantes/toxicidad , Tensoactivos/toxicidad , Animales , Bovinos , Córnea/metabolismo , ADN/metabolismo , Epitelio/metabolismo , FN-kappa B/metabolismo , Técnicas de Cultivo de Órganos , Tensoactivos/farmacocinética , Factor de Transcripción AP-1/metabolismoRESUMEN
OBJECTIVES: To evaluate the efficacy of systemic or topical administration of cyclosporin A (CsA) after limbal transplantation of stem cell allografts in rabbits. METHODS: Thirty-six rabbits underwent corneal epithelial debridement and limbal ablation to induce ocular surface disease and were then treated by limbal allograft transplantation. Animals received either systemic CsA (10 mg/kg per day, intramuscularly), 1% CsA eyedrops, or vehicle eyedrops immediately after transplantation and 28 days thereafter. Concentration of CsA in plasma and aqueous humor was determined by fluorescence polarization immunoassay after 4 weeks of therapy. Graft survival was inspected clinically. RESULTS: Both systemic and topical administration of CsA resulted in a significant prolongation of graft survival. In addition, one of seven of the limbal allografts in either group of systemic and topical CsA survived >60 days on cessation of CsA. There was no significant difference in mean survival time between systemic and topical application, although plasma levels of CsA were significantly higher after systemic administration. However, a significant higher aqueous concentration was found in topical treatment. CONCLUSIONS: Limbal allografts were stable in maintaining the reconstructed ocular surface under attentive postoperative immunosuppression. Topically administered CsA was as effective as systemic use.
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Trasplante de Células , Enfermedades de la Córnea/cirugía , Ciclosporina/administración & dosificación , Supervivencia de Injerto/efectos de los fármacos , Inmunosupresores/administración & dosificación , Limbo de la Córnea/citología , Trasplante de Células Madre , Animales , Humor Acuoso/metabolismo , Enfermedades de la Córnea/tratamiento farmacológico , Enfermedades de la Córnea/patología , Ciclosporina/farmacocinética , Modelos Animales de Enfermedad , Células Epiteliales/trasplante , Estudios de Seguimiento , Inmunosupresores/farmacocinética , Inyecciones Intramusculares , Limbo de la Córnea/metabolismo , Masculino , Soluciones Oftálmicas/administración & dosificación , Conejos , Trasplante Homólogo , Resultado del TratamientoRESUMEN
Our purpose was to investigate the relationship between insufficient tear secretion and decreased corneal sensitivity. We studied 59 patients with dry eye, 15 of whom had Sjögren's syndrome (SS), and 26 healthy subjects. Corneal sensitivity was measured by the esthesiometer of Cochet and Bonnet. Schirmer test with or without anesthesia, tear clearance rate, tear function index, and rose bengal and fluorescein staining were also evaluated. The mean corneal sensitivity of either dry-eye group (4.6 +/- 1.2 and 4.5 +/- 1.2 cm for non-SS and SS dry eye, respectively) was significantly lower than that of the control (5.8 +/- 0.4 cm, p < 0.001). Corneal sensitivity correlated significantly with the Schirmer values with anesthesia and the tear function index in the two dry-eye groups and the control (p < 0.05). There were significant relationships between corneal sensitivity and the rose bengal and fluorescein scores in the three groups (p < 0.05). Hyposecretion of tears in dry eye may lead to pathologic changes in corneal epithelium and a decline in corneal sensitivity. Prompt treatment of dry eye is essential to maintain a normal corneal protective mechanism.
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Córnea/fisiopatología , Síndromes de Ojo Seco/fisiopatología , Lágrimas/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Valores de ReferenciaRESUMEN
OBJECTIVE: To investigate whether reflex tearing measured by the Schirmer test with nasal stimulation is better correlated with lymphocytic infiltration of the lacrimal gland and is thus a more appropriate test for Sjögren's syndrome (SS). METHODS: 272 patients with dry eye (age 55.5 +/- 13.5 years) with regular Schirmer test results < 10 mm were recruited for the study. They were divided into 2 groups depending on the results of the Schirmer test with nasal stimulation. 24 age and sex matched patients from each group were selected for lacrimal gland biopsy and 10 patients for salivary gland biopsy, and the levels of systemic autoantibodies and degrees of ocular surface staining and lymphocytic infiltration of the lacrimal and salivary glands were compared. RESULTS: Of the 272 patients, 187 (68.8%) had good reflex tearing (GT) and 85 (31.2%) had poor reflex tearing (PT). There were no significant differences in the age or sex of the 2 groups. Lymphocytic infiltration of the lacrimal gland was observed in 7 GT cases and 22 PT cases out of 24 cases. Lymphocytic infiltration of the salivary gland was also higher in the PT (6 of 10) than the GT group (2 of 10). CONCLUSION: Evaluation of reflex tearing by the Schirmer test with nasal stimulation identifies 2 groups of patients with dry eye. Those with poor reflex tearing were more likely to have autoantibodies and lymphocytic infiltration of the exocrine glands consistent with SS.
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Movimiento Celular , Aparato Lagrimal/fisiopatología , Linfocitos/fisiología , Reflejo , Síndrome de Sjögren/diagnóstico , Lágrimas , Adulto , Anciano , Biopsia , Femenino , Humanos , Aparato Lagrimal/diagnóstico por imagen , Aparato Lagrimal/patología , Masculino , Persona de Mediana Edad , Radiografía , Glándulas Salivales/patologíaRESUMEN
OBJECTIVE: To determine the diagnostic value in Sjögren's syndrome (SS) of evaluating the presence of focal adenitis in labial salivary gland (LSG) and lacrimal gland (LG) biopsy specimens. METHODS: We studied 105 cases with suspected SS and performed biopsies prospectively. The diagnostic criteria of Fox, et al were followed. The Chisholm-Mason grading standard and Greenspan focus score were used for evaluation. RESULTS: Epimyoepithelial islands and severe lymphocyte infiltration with germinal centers were observed only in LG biopsy specimens (5.9 and 3.5%, respectively). The lymphocytic focus scores of LG (2.5 +/- 2.1/4 mm2) were significantly higher than those of LSG specimens (1.3 +/- 1.1/4 mm2) in 23 cases with both biopsy results (p < 0.05). Six of the 32 LSG biopsies were normal while their LG biopsies were significantly higher. Conversely, 5 of the 32 LG biopsies were aided by corresponding LSG biopsies, since in LG biopsies no glandular tissue was obtained. Evaluation of both specimens for the same patient was significantly more effective in diagnosing SS than using either specimen alone (p < 0.01). CONCLUSION: LG biopsy specimens have a more evident histopathology than LSG biopsy specimens. We recommend that both LSG and LG biopsies be performed in patients with suspected SS to reduce false negative results and improve diagnostic accuracy.
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Aparato Lagrimal/patología , Labio/patología , Glándulas Salivales/patología , Síndrome de Sjögren/diagnóstico , Biopsia , Demografía , Epitelio/patología , Humanos , Hiperplasia/patología , Metaplasia/patología , Serología , Síndrome de Sjögren/patología , LágrimasRESUMEN
BACKGROUND: The study sought to determine dynamic changes and theoretical bases of a clinical diagnostic test, the tear clearance rate. METHODS: Thirty four healthy subjects ranging in age from 22 to 84 years underwent examination of tear clearance rate, the Schirmer test with anaesthesia, as well as fluorophotometric measurement of tear turnover, tear volume, and tear flow. By applying 0.5% fluorescein into the conjunctival sac and subsequently measuring colour fades on a Schirmer strip, the tear clearance rate for assessing tear drainage was divided into nine grades. The results of the tear clearance rate were compared with those of the basal tear turnover and tear flow obtained from fluorophotometry. RESULTS: Significant relations were found between the tear clearance rate and the basal tear turnover or tear flow (r = 0.91 and 0.79, respectively, p = 0.0001). Considering the grades of progression from low to high, each grade of tear clearance rate showed a 12.5% increase in basal tear turnover (3.59%/min) and tear flow (0.38 microliter/min). There was no significant correlation between age and the basal tear turnover, tear volume, tear flow, or the tear clearance rate. CONCLUSION: The tear clearance rate is proposed as a simple and useful way to estimate basal tear turnover and tear flow, and measure tear drainage indirectly.
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Lágrimas/fisiología , Adulto , Anciano , Anciano de 80 o más Años , Fluorofotometría , Humanos , Persona de Mediana Edad , Lágrimas/metabolismoRESUMEN
BACKGROUND: The study sought to investigate the histogenesis of retinoblastoma. METHODS: One hundred specimens of retinoblastomas were examined along with those of 18 astrocytic gliomas and 15 medulloblastomas to compare similarities of glial differentiation in retinoblastoma and the two types of brain tumour. Employing avidin-biotin immunoperoxidase technique, antibodies were applied against neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP), and S-100 protein (S-100). RESULTS: Most rosettes and fleurettes, and some undifferentiated cells in retinoblastomas were NSE positive, but GFAP and S-100 negative. GFAP and S-100 positive cells in retinoblastomas were detected mostly in well differentiated glial cells which were interpreted as reactive or non-neoplastic cells. Some of the GFAP and S-100 positive cells in retinoblastomas were defined as tumour cells that resembled neoplastic astrocytes in astrocytic gliomas and medulloblastomas. CONCLUSION: Retinoblastoma may arise from primitive bipotential or multipotential cells capable of neuronal and glial differentiation.
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Neoplasias del Ojo/patología , Neuroglía/patología , Neuronas/patología , Retinoblastoma/patología , Astrocitoma/patología , Transformación Celular Neoplásica/metabolismo , Neoplasias Cerebelosas/patología , Preescolar , Neoplasias del Ojo/metabolismo , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Humanos , Técnicas para Inmunoenzimas , Masculino , Meduloblastoma/patología , Fosfopiruvato Hidratasa/metabolismo , Retinoblastoma/metabolismo , Proteínas S100/metabolismoRESUMEN
PURPOSE: The present study was performed to culture human fetal ova to determine whether they can be matured and cryopreserved using ultrarapid freezing. METHODS: Thirty-three pairs of fetal ovaries were obtained from fetuses of 16-20 weeks' gestation following elective abortion. Ovarian tissues were minced into approximately 1-mm sizes and cultured in Waymouth media either before or after ultrarapid freezing. The Waymouth medium was supplemented with 15% (v/v) fetal bovine serum, 0.03 IU/ml FSH and 35 ng/ml insulin. The tissue was cultured at 37 degrees C in 5% CO2 in air for 5-25 days in Falcon dishes and 30-40 days in Costar Transwell-COL membranes prior to induction of final maturation in the presence of LH and human follicular fluid. Minced tissues were also frozen by ultrarapid freezing in M199 with 4.2 M dimethylsulfoxide (DMSO) and 0.35 M sucrose and plunged directly into liquid nitrogen. For thawing, the straws were plunged into a 37 degrees C water bath for 5 s. The contents were then expelled and diluted 1:5 with thawing medium containing 0.42 M sucrose. After washing the thawed tissues were cultured as described for the fresh tissues. RESULTS: Patches of monolayer consisting of fibroblasts had formed within 2-3 days of culture of fresh tissues. After 1 week of culture, follicles separated out from the ovarian tissue but remained attached to the monolayer. The maximal number of follicles separating out from the tissue appeared about 1 week after initiating the culture (154 follicles per 10 fields at Day 5 and 61 and Day 25). After 40 days of culture in Costar dishes, 34% of the ova reached a diameter of more than 80 microns, which was significantly higher than at the beginning of culture (6%; P < 0.05). Among these ova, 34% were found to be surrounded by the zona pellucida, which was not observed at the beginning of culture. Following induction of final maturation, extrusion of the first polar body was noted in 25% of ova grown in Costar dishes for 40 days. Twelve percent of the oocytes showed the first polar body when they were grown in Costar dishes for less than 30 days. For frozen-thawed tissues, 14% of minced ovarian tissues displayed central necrosis immediately after thawing. Following digestion and Trypan blue staining, 75% of ova and 85% of somatic cells survived ultrapid freezing. Nineteen percent of the ova which have been cultured as described for fresh tissues displayed extrusion of the first polar body, comparing favorably with the 25% maturation rate observed with the fresh tissue (P > 0.05). CONCLUSION: This study demonstrates that morphologically normal, mature human ova can be obtained from primordial follicles in vitro development. Using a simple, quick ultrarapid freezing method, human fetal ova can be cryopreserved in the form of minced tissue without significantly compromising their ability to grow in vitro.
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Criopreservación/métodos , Ovario/citología , Ovario/embriología , Óvulo/crecimiento & desarrollo , Animales , Bovinos , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Células Cultivadas , Crioprotectores/análisis , Crioprotectores/farmacología , Medios de Cultivo/análisis , Medios de Cultivo/farmacología , Dimetilsulfóxido/análisis , Dimetilsulfóxido/farmacología , Femenino , Hormona Folículo Estimulante/análisis , Hormona Folículo Estimulante/farmacología , Líquido Folicular/fisiología , Humanos , Insulina/análisis , Insulina/farmacología , Hormona Luteinizante/farmacología , Oocitos/citología , Oocitos/efectos de los fármacos , Oocitos/fisiología , Oogénesis/fisiología , Óvulo/citología , Óvulo/efectos de los fármacos , Albúmina Sérica Bovina/análisis , Albúmina Sérica Bovina/farmacología , Zona Pelúcida/fisiologíaRESUMEN
OBJECTIVE: To develop an efficient way to evaluate tear dynamics clinically. PARTICIPANTS: Three hundred fifty-two patients with dry eye, 64 of whom had Sjögren syndrome, and 55 normal subjects. DESIGN: Because various forces that affect tear drainage are reflected in the values of the Schirmer test with anesthesia and the tear clearance rate, we introduced a new measure of tear dynamics, the tear function index, which is the value obtained from dividing the value of the Schirmer test with anesthesia by the tear clearance rate. RESULTS: The tear function index was more specific (91.8%) and sensitive (78.9%) in diagnosing dry eye associated with Sjögren syndrome than was the Schirmer or tear clearance rate test alone. Tear function indexes below 96 were consistent with dry eye and those below 34 were seen primarily in patients with Sjögren syndrome. CONCLUSIONS: The tear function index offers a new method to evaluate tear production with the Schirmer test, introduces an extended way to measure tear flow combining with tear drainage, and gives a practical measure to diagnose dry eye.
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Síndromes de Ojo Seco/diagnóstico , Lágrimas/fisiología , Síndromes de Ojo Seco/fisiopatología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sensibilidad y EspecificidadRESUMEN
The classical concept of sex determination in mammals is that a Y chromosomal gene controls the development of the indifferent gonad into a testis. Subsequent divergence of sexual phenotypes is secondary to this gonadal determination. The most likely candidate gene is SRY (sex-determining region Y) in humans, and Sry in mouse. However, several lines of evidence indicate that sexual dimorphism occurs even before the indifferent gonad appears. Here we present evidence that bovine male embryos generally develop to more advanced stages than do females during the first 8 days after insemination in vitro. Corresponding relationships between both cell numbers and mitotic indices and sex were also seen. Although it is not clear whether this phenomenon involves factors originating before or after fertilization, these findings suggest that sex-related gene expression affects the development of embryos soon after activation of the embryonic genome and well before gonadal differentiation.
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Desarrollo Embrionario y Fetal , Caracteres Sexuales , Animales , Bovinos , Técnicas de Cultivo , Femenino , Fertilización In Vitro , Cariotipificación , MasculinoRESUMEN
The ovaries of two infertile cows of high breeding value were recovered after slaughter, and a total of 222 oocytes were obtained. Of these, 156 were classified as of good or fair quality and were subjected to in vitro maturation, in vitro fertilisation (using frozen semen from three bulls of high breeding value) and in vitro culture procedures. After eight days, 27 embryos were obtained, of which 13 were transferred fresh, and 14 were frozen. Three recipients of fresh embryos became pregnant; two calved and one aborted at four months. One of eight recipients of frozen-thawed embryos became pregnant but aborted at three months.
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Bovinos , Transferencia de Embrión/veterinaria , Fertilización In Vitro/métodos , Mataderos , Animales , Criopreservación/veterinaria , Transferencia de Embrión/métodos , Femenino , Linaje , Preservación de Semen/veterinariaRESUMEN
A co-culture system using a suspension of detached bovine oviducal epithelial cells (BOEC) has been developed as an effective culture method for supporting the development of bovine embryos derived from oocytes matured and fertilized in vitro. Four commercially available culture media (Waymouth's, Ham's F-10, TCM 199 and Ménézo's B2) supplemented with 10% oestrous cow serum, and a modified Tyrode's medium (TALP) supplemented with 0.6% bovine serum albumin were used. Ménézo's B2 resulted in the highest percentages of total uncleaved presumptive zygotes, and of the cleaved zygotes that reached at least the morula stage (31-46% and 66-74%, respectively). The embryos produced in vitro in B2 with BOEC resembled embryos produced in vivo with regard to numbers of cells (averaging 45.4 in morulae, 101.5 in blastocysts, 174.7 in hatching blastocysts and 195.9 in hatched blastocysts), rate of development (hatching on Day 8-9 of culture in vitro), rate of hatching (66% of cleaved zygotes) and pregnancy rates (63%) resulting from the transcervical transfer of selected embryos.
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Bovinos/fisiología , Embrión de Mamíferos/fisiología , Desarrollo Embrionario y Fetal/fisiología , Trompas Uterinas/fisiología , Fertilización In Vitro/métodos , Viabilidad Fetal/fisiología , Animales , Blastocisto/citología , Células Cultivadas , Transferencia de Embrión , Células Epiteliales , Trompas Uterinas/citología , Femenino , EmbarazoRESUMEN
The male:female ratio of developing bovine embryos produced and allowed to develop in vitro and in vivo was determined retrospectively from the cytogenetic analysis of 804 embyos. The overall male:female ratio of the 307 (38%) embryos that could be sexed was 162 (52.8%):145 (47.2%) and did not differ (P>0.05) from the expected 1:1 ratio. Among premorula stage embryos produced in vivo (n = 66) and in vitro (n = 30), the ratios were 1.2:1 and 0.76:1, respectively. Among morulae and blastocysts produced in vivo (n = 74), produced and cultured in vitro (n = 106, and produced in vitro and cultured in vivo (n = 31), the ratios were 1.11:1, 1.3:1 and 0.94:1, respectively, none of which differed significantly from 1:1. There was no difference (P>0.05) in the number of cells or mitotic index between male and female morulae and blastocyst, respectively.
RESUMEN
Two culture techniques, an open system using 1.0-ml medium and a covered system using 50-mul droplets of medium covered by paraffin oil, used for in vitro bovine follicular oocyte maturation were compared. Estradiol-17beta (E2) was added to Ham's F-10 medium together with other supplementations. In the absence of oocytes, E2 concentration remained unchanged in the 1.0-ml open system, but it decreased gradually in the 50-mul covered system. In the presence of oocytes, E2 increased significantly in the 1.0-ml open system, but this increase could not be detected in the 50-mul covered system. Progesterone (P4) concentration increased in both systems, but it was much higher in the 1.0-ml culture than in the 50-mul culture. The two culture systems allowed an identical nuclear oocyte maturation rate of 88.6 vs 87.0%, a sperm penetration rate of 54.3 vs 59.6%, and a polyspermy rate of 6.8 vs 11.6% for 1.0-ml and 50-mul systems, respectively. The cleavage rate, however, differed significantly (78.3 vs 41.7% for 1.0-ml and 50-mul systems, respectively). It is concluded that diffusion of steroids into the paraffin oil occurs and may affect the cleavage rate but not the maturation or penetration rate.