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2.
Environ Pollut ; 261: 114007, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32036198

RESUMO

Zearalenone (ZEA), an estrogen-like mycotoxin, is commonly detected in animal feeds including improperly stored grains. It has been well demonstrated that ovarian granulosa cells (GCs) perform vital roles during follicular development, however, the competing endogenous RNA (ceRNA) network in GCs after ZEA exposure remains to be well described. Here, for the first time, we adopted whole-transcriptome sequence technology to explore the molecular mechanism of ZEA toxicology on porcine GCs. The results provide evidence that the cell cycle of porcine GCs is arrested in the G2/M phase after exposure to ZEA. Furthermore, bioinformation analysis found that cell cycle arrest related genes were perturbed, including CDK1, CCNB1, CDC25A, and CDC25C, which was consistent with the results of RT-qPCR, immunofluorescence, and Western Blotting. Based on the whole-transcriptome sequence data, by constructing ceRNA networks related to cell cycle arrest, we observed that ZEA exposure arrested cell cycle progression at the G2/M phase in porcine GCs, and non-coding RNAs (ncRNAs) played an important role in this process via regulating the expressions of cell cycle arrest related genes. Taken together, our data here provides strong data to support that the toxicological mechanism regarding the widely distributed toxicant ZEA acts through ceRNA networks in porcine granulosa cells.


Assuntos
Zearalenona , Animais , Células Cultivadas , Feminino , Perfilação da Expressão Gênica , Células da Granulosa , RNA , Suínos
3.
Am J Gastroenterol ; 115(5): 653-661, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31464742

RESUMO

INTRODUCTION: The objective was to evaluate diagnostic performance of multiple methods used to assess gastric tube placement verification in neonates, infants, and children. METHODS: A systematic review using the methods outlined in the Cochrane Handbook for Reviews of Diagnostic Test Accuracy was conducted. Eight databases were searched. Studies on neonates, infants, and children in which researchers compared different methods for gastric tube placement verification with x-ray reference standard were eligible in the review. RESULTS: Eight studies involving 911 participants that evaluated 9 index tests for gastric tube placement verification were included. Most studies were of moderate methodological quality, and most index tests were assessed in small individual studies. pH testing with cutoff values ≤ 6 for gastric tube position confirmation was the only index test subjected to meta-analysis, with the summary sensitivity and specificity being 0.77 (95% confidence interval [CI] 0.56-0.90) and 0.42 (95% CI 0.16-0.73). Other tests for gastric tube placement verification showed great variations in sensitivities and specificities. DISCUSSION: pH ≤ 6 is not sufficiently accurate to be recommended for gastric tube placement verification in neonates, infants, and children. Diagnostic performance of pH ≤ 4 or 5 and other methods cannot be determined because of the paucity of data and methodological variations in studies. Clinical practice related to the diagnostic tests used will continue to be dictated by local preferences and cost factors, until stronger evidence becomes available.


Assuntos
Intubação Gastrointestinal/normas , Estômago/química , Estômago/diagnóstico por imagem , Auscultação , Capnografia , Pré-Escolar , Humanos , Concentração de Íons de Hidrogênio , Lactente , Recém-Nascido , Radiografia , Ultrassonografia
4.
J Cell Physiol ; 234(10): 18214-18229, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30859584

RESUMO

Previous studies have shown that primordial germ cell-like cells (PGCLCs) can be obtained from human, porcine and mouse skin-derived stem cells (SDSCs). In this paper, we found retinoic acid (RA), the active derivative of vitamin A, accelerated the growth of porcine primordial germ cells (pPGCs) and porcine PGCLCs (pPGCLCs) which were derived from porcine SDSCs (pSDSCs). Moreover, flow cytometry results revealed that the proliferation promoting effect of RA was attenuated by U0126, a specific inhibitor of extracellular signal-regulated kinase (ERK). Western blot analysis showed the protein level of ERK, phosphorylated ERK, cyclin D1 (CCND1), and cyclin-dependent kinase 2 (CDK2) increased after stimulation with RA, and this effect could also be abolished by U0126. Our data revealed that ablation of ERK expression by U0126 should significantly decrease proliferation of pPGCLCS. This reduction was because CCND1 and CDK2 proteins level decrease and subsequently the pPGCLCs were arrested in the G0/G1 phase. In addition, we also confirmed RA indeed promoted the proliferation of pPGCs isolated from porcine fetal genital ridges in vitro. Furthermore, our data indicated that DNA methylation pattern were changed in pPGCLCs and this pattern were more similar to pPGCs.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Tretinoína/farmacologia , Animais , Células Cultivadas , Ciclina D1/metabolismo , Quinase 2 Dependente de Ciclina/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fase G1/efeitos dos fármacos , Células Germinativas/metabolismo , Fosforilação/efeitos dos fármacos , Fase de Repouso do Ciclo Celular/efeitos dos fármacos , Células-Tronco/metabolismo , Suínos
5.
J Agric Food Chem ; 67(9): 2679-2690, 2019 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-30650308

RESUMO

The mycotoxin ochratoxin A (OTA), a naturally occurring food contaminant, has a toxic effect on the growth and development of follicles in pigs. However, little is known regarding the specific toxic effects of OTA exposure on oocytes and granulosa cells (GCs). In this study, we cultured porcine ovarian GCs and exposed them to OTA in vitro in order to explore the mechanism causing the negative effects. Initially, it was found that OTA exposure inhibited cell viability in a time and dose dependent manner. We also showed that OTA exposure increased oxidative stress, decreased proliferation ratio, and increased apoptosis ratio in GCs. We revealed an important role for the PI3K/AKT signal pathway in GC proliferation and apoptosis by RNA-seq analysis. The results not only showed that OTA treatment significantly affected the expression of genes within the PI3K/AKT pathway but also demonstrated a concrete relationship between the PI3K/AKT pathway and GC cell proliferation and apoptosis. In conclusion, the results demonstrated that OTA exposure impaired porcine GC growth via the PI3K/AKT signaling pathway.


Assuntos
Células da Granulosa/fisiologia , Ocratoxinas/toxicidade , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Suínos , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Expressão Gênica/efeitos dos fármacos , Glutationa/análise , Células da Granulosa/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos
6.
Aging (Albany NY) ; 10(11): 3486-3506, 2018 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-30472698

RESUMO

Zearalenone (ZEA) is a well-known exogenous endocrine disruptor and can lead to severe negative effects on the human and animal reproductive process. Using a follicle culture model, we have previously shown that ZEA exposure significantly affected the follicular development and antrum formation but the underlying mechanisms are not well known. Therefore, in this study, we explored the metabolomic changes of granulosa cell (GC) culture media with or without ZEA exposure. The results showed that ZEA significantly increased phosphatidylcholine or phosphatidyl ethanolamine adducts in culture medium. A comprehensive analysis with the metabolome data from follicular fluid of small and large antral follicles showed that lyso phosphatidylcholine (LPC) was accumulated during follicle growth, but was depleted by ZEA exposure. Exogenous supplement with LPC to the follicle growth media or oocyte maturation media can partly protect the defect of ZEA exposure on follicular antrum formation and oocyte maturation. Taken together, our results demonstrate that ZEA exposure hinders the follicular growth and exogenous LPC can practically protect the defect of ZEA on follicular development and oocyte maturation.


Assuntos
Oócitos/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Fosfatidilcolinas/farmacologia , Zearalenona/toxicidade , Animais , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/fisiologia , Fator de Ativação de Plaquetas/metabolismo , Suínos
7.
Toxicol Appl Pharmacol ; 356: 191-203, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30138655

RESUMO

Zearalenone (ZEA) is one of mycotoxins which are from corn, sorghum and wheat. As an estrogenic compound, ZEA mainly affects animal growth and reproduction with causing abnormal reproduction capability. Previous studies have shown that ZEA poses adverse effects on follicular development, but the mechanism of genetic toxicity of ZEA is not understood. The purpose of this study was to explore the effects of ZEA exposure on granulosa cells which play vital roles during follicular development. Mouse granulosa cells were exposed to 10 µM or 30 µM ZEA for 72 h in vitro, and the differences in gene expression patterns between control and ZEA exposures were analyzed by RNA-seq. The data demonstrated that 30 µM ZEA had a significant effect on the gene expression, especially ZEA exposure increased the expression of many genes related to different kinds of cancers and cancer related pathways like Hippo signaling pathway and the related genes, such as Ccnd1, Smad3, Tead3, Yap1 and Wwtr1. Furthermore, immunohistochemistry confirmed the increase in the protein levels of YAP1, WWTR1 and CCND1 in 30 µM ZEA exposure group. Collectively, this investigation indicated that ZEA exposure promoted the expression of tumorigenesis genes in mouse granulosa cells to.


Assuntos
Carcinógenos/toxicidade , Genes Neoplásicos/efeitos dos fármacos , Células da Granulosa/efeitos dos fármacos , Micotoxinas/toxicidade , Neoplasias Ovarianas/induzido quimicamente , Neoplasias Ovarianas/genética , Ovário/citologia , Zearalenona/toxicidade , Animais , Carcinogênese , Transformação Celular Neoplásica/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Camundongos , Ovário/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos
8.
Int J Biol Sci ; 14(3): 294-305, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29559847

RESUMO

Zearalenone (ZEA), a metabolite of Fusarium fungi, is commonly found on moldy grains. Because it can competitively combine to estrogen receptor to disrupt estrogenic signaling, it has been reported to have serious adverse effects on animal reproduction systems. In order to explore the genotoxic effects of ZEA exposure on ovarian somatic cells, porcine granulosa cells were exposed to 10 µM and 30 µM ZEA for 24 or 72 h in vitro. The results showed that ZEA exposure for 24 h remarkably reduced the proliferation of porcine granulosa cells in a dose-dependent manner as determined by MTT analysis and flow cytometry. Furthermore, exposure to ZEA for 72 h induced apoptosis, and RNA sequence analysis also revealed that the expression of apoptosis related genes were altered. RT-qPCR, immunofluorescence and western blot analysis further confirmed the expression of DNA damage and repair related genes (γ-H2AX, BRCA1, RAD51 and PRKDC) were increased in ZEA exposed granulosa cells. When the estrogen antagonist, tamoxifen, was added with ZEA in the culture medium, the DNA damage and repairment by ZEA returned to normal level. Collectively, these results illustrate that ZEA disrupts genome stability and inhibits growth of porcine granulosa cells via the estrogen receptors which may promote granulosa cell apoptosis when the DNA repair system is not enough to rescue this serious damage.


Assuntos
Estrogênios não Esteroides/toxicidade , Instabilidade Genômica/efeitos dos fármacos , Células da Granulosa/efeitos dos fármacos , Zearalenona/toxicidade , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Western Blotting , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Meios de Cultura , Dano ao DNA/genética , Reparo do DNA/genética , Relação Dose-Resposta a Droga , Feminino , Células da Granulosa/citologia , Células da Granulosa/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de RNA , Suínos
9.
World J Gastroenterol ; 23(1): 87-92, 2017 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-28104983

RESUMO

AIM: To explore the effect of hydrogen sulfide (H2S) on restraint water-immersion stress (RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate (ATP)-sensitive potassium (KATP) channels and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway on such an effect. METHODS: Male Wistar rats were randomly divided into a control group, a physiological saline (PS) group, a sodium hydrosulfide (NaHS) group, a glibenclamide (Gl) group, Gl plus NaHS group, a pyrrolidine dithiocarbamate (PDTC) group, and a PDTC plus NaHS group. Gastric mucosal injury was induced by RWIS for 3 h in rats, and gastric mucosal damage was analyzed after that. The PS, NaHS (100 µmol/kg body weight), Gl (100 µmol/kg body weight), Gl (100 µmol/kg or 150 µmol/kg body weight) plus NaHS (100 µmol/kg body weight), PDTC (100 µmol/kg body weight), and PDTC (100 µmol/kg body weight) plus NaHS (100 µmol/kg body weight) were respectively injected intravenously before RWIS. RESULTS: RWIS induced serious gastric lesions in the rats in the PS pretreatment group. The pretreatment of NaHS (a H2S donor) significantly reduced the damage induced by RWIS. The gastric protective effect of the NaHS during RWIS was attenuated by PDTC, an NF-κB inhibitor, and also by glibenclamide, an ATP-sensitive potassium channel blocker, in a dose-dependent manner. CONCLUSION: These results suggest that exogenous H2S plays a protective role against RWIS injury in rats, possibly through modulation of KATP channel opening and the NF-κB dependent pathway.


Assuntos
Mucosa Gástrica/efeitos dos fármacos , Sulfeto de Hidrogênio/farmacologia , Canais KATP/metabolismo , NF-kappa B/metabolismo , Animais , Mucosa Gástrica/lesões , Glibureto/administração & dosagem , Glibureto/farmacologia , Sulfeto de Hidrogênio/administração & dosagem , Injeções Intravenosas , Canais KATP/antagonistas & inibidores , Masculino , NF-kappa B/antagonistas & inibidores , Pirrolidinas/administração & dosagem , Pirrolidinas/farmacologia , Ratos , Ratos Wistar , Transdução de Sinais , Estresse Psicológico/complicações , Tiocarbamatos/administração & dosagem , Tiocarbamatos/farmacologia
10.
Arch Toxicol ; 91(3): 1279-1292, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27405655

RESUMO

Di (2-ethylhexyl) phthalate (DEHP) is a plasticizer which is widely used in the manufacture of plastics. As a common environmental contaminant and recognized endocrine disrupting chemical, DEHP is able to deregulate the functions of a variety of tissues, including the reproductive system both in males and females. In order to investigate the possible effects of DEHP on the first wave of folliculogenesis, occurring in the mouse ovary postnatally, mice were administered 20 or 40 µg/kg DEHP through intraperitoneal injection at days 5, 10 and 15 post partum (dpp). Following DEHP treatment the gene expression profile of control and exposed ovaries was compared by microarray analyses at 20 dpp. We found that in the exposed ovaries DEHP significantly altered the transcript levels of several immune response and steroidogenesis associated genes. In particular, DEHP significantly decreased the expression of genes essential for androgen synthesis by theca cells including Lhcgr, Cyp17a1, Star and Ldlr. Immunohistochemistry and immune flow cytometry confirmed reduced expression of LHCGR and CYP17A1 proteins in the exposed theca cells. These effects were associated to a significant reduction in ovarian concentrations of progesterone, 17ß-estradiol and androstenedione along with a reduction of LH in the serum. Although we did not find a significant reduction of the number of primary, secondary or antral follicles in the DEHP exposed ovaries when compared to controls, we did observe that theca cells showed an altered structure of the nuclear envelope, fewer mitochondria, and mitochondria with a reduced number of cristae. Collectively, these results demonstrate a deleterious effect of DEHP exposure on ovarian steroidogenesis during the first wave of folliculogenesis that could potentially affect the correct establishment of the hypothalamic-pituitary-ovarian axis and the onset of puberty.


Assuntos
Dietilexilftalato/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Esteroides/metabolismo , Animais , Feminino , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/ultraestrutura , Camundongos Endogâmicos , Análise de Sequência com Séries de Oligonucleotídeos , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Ovário/efeitos dos fármacos , Ovário/fisiologia , Puberdade
11.
Int J Gynecol Cancer ; 21(8): 1357-65, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21720253

RESUMO

OBJECTIVE: The objectives of the study were to evaluate the role of mitogen-activated protein kinase (MAPK) signaling in normal, hyperplastic, and neoplastic endometrium in relation to estrogen receptor (ER) status and to investigate whether 17ß-estradiol (E2) and tamoxifen (TAM) mediate the proliferation and apoptosis of endometrial cancer cells through the MAPK pathway. METHODS: The expressions of phosphorylated and total extracellular signal-regulated kinases 1/2 (phosphorylated extracellular signal-regulated kinase 1/2 [p-ERK1/2] and total ERK1/2 [t-ERK1/2]) were analyzed with immunohistochemistry in normal, hyperplastic, and neoplastic endometrium. The expression levels of p-ERK1/2 and t-ERK1/2 in RL95-2 and KLE after stimulation by E2, progesterone (P), and TAM were detected by Western blotting. The effects of E2 and TAM in combination with MAPK pathway inhibitors on the growth and apoptosis of endometrial cancer cells were examined by the MTS assay and flow cytometry analysis. RESULTS: The expression level of p-ERK1/2 was significantly associated with the International Federation of Gynecology and Obstetrics stage (P = 0.0072). The ratio of phosphorylated/total ERK1/2 was higher in ER-positive endometrial cancer tissues and cells (P < 0.05). 17ß-Estradiol increased ERK1/2 phosphorylation, and TAM decreased ERK1/2 phosphorylation in endometrial cancer cell lines within 30 minutes (P < 0.05). The MEK1/2 inhibitor, U0126, and the stress-activated protein kinase/c-Jun NH2-terminal kinase inhibitor, SP600125, significantly suppressed the proliferation of human endometrial cancer cell lines RL95-2 and KLE induced by E2 (P < 0.05). The level of TAM-induced apoptosis was greater in KLE than in RL95-2 cells, and the p38 cascade was involved in the TAM-induced apoptosis of both cell lines (P < 0.05). CONCLUSIONS: The cross-talk between MAPK signaling and ER status might exert a key role in progression of endometrial cancer. Furthermore, the effects of E2 or TAM on the proliferation or apoptosis of ER-positive and ER-negative endometrial cancer cells were mediated through distinct MAPK pathways. These mechanisms might contribute to ER-specific differences in MAPK activation for molecular-target therapies in endometrial carcinoma.


Assuntos
Adenocarcinoma/etiologia , Neoplasias do Endométrio/etiologia , Sistema de Sinalização das MAP Quinases , Receptor Cross-Talk , Receptores de Estrogênio/metabolismo , Adenocarcinoma/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose , Estudos de Casos e Controles , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Neoplasias do Endométrio/metabolismo , Estradiol , Feminino , Humanos , Pessoa de Meia-Idade , Fosforilação , Tamoxifeno
12.
Cancer Sci ; 101(7): 1618-23, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20487263

RESUMO

Methylation is an important silencing mechanism of breast and ovarian cancer susceptibility gene 1 (BRCA1) expression in sporadic ovarian cancer. However, the role of BRCA1 methylation in chemotherapy in sporadic ovarian cancer and the related pathways have not been understood completely. This study has determined the roles of BRCA1 hypermethylation in chemotherapy of sporadic ovarian cancer and its related signaling pathways. We used bisulfite sequencing, real-time polymerase chain reaction, and western blotting to check the methylation state and expression levels of BRCA1 of the following cell lines: platinum-sensitive human ovarian cancer cell line COC1, platinum-resistant cell line COC1/DDP, SKOV-3, and 5-Aza-dC treated COC1. The cisplatin sensitivity of ovarian cancer cells was examined by MTS (methyl-thiazol tetrazolium) assay. Tumorigenicity in vivo and DDP-based chemosensitivity were compared among the above cells. Phosphatidylinositol 3'-kinase (PI3K)-Akt pathway activation in ovarian cancer cells was studied by western blotting. The frequency of BRCA1 methylation in the COC1 cell line was higher than in COC1/DDP and SKOV-3 cell lines, whereas the mRNA and protein expression of BRCA1 were lower than in the COC1/DDP and SKOV-3 cell lines. DNA demethylation decreased the chemosensitivity of COC1 cells and partially increased the expression levels of BRCA1. The activation of the PI3K-Akt pathway was low in ovarian cancer cells. Our results indicate that hypermethylation of BRCA1 might play an important role in the chemosensitivity of ovarian cancer, and that the PI3K-Akt pathway is not involved in this response.


Assuntos
Neoplasias da Mama/genética , Metilação de DNA , Genes BRCA1 , Predisposição Genética para Doença , Neoplasias Ovarianas/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Linhagem Celular Tumoral , DNA de Neoplasias/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Proteínas Supressoras de Tumor/genética
13.
Pain Med ; 11(10): 1564-75, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21199306

RESUMO

OBJECTIVE: The objective of this study was to compare the effects of Sanyinjiao (SP6), Xuanzhong (GB39), and an adjacent non-meridian point on menstrual pain and uterine arterial blood flow in primary dysmenorrhea patients. DESIGN: The design of the study was a prospective, randomized controlled trial. SETTING: The setting of the study was the Shandong Institute of Medical Imaging, Jinan, China. PATIENTS: The patients were 52 women with primary dysmenorrhea. INTERVENTIONS: Women received electroacupuncture (EA) at SP6 (n=13), GB39 (n=14), and an adjacent non-meridian point (n=12), respectively, for 10 minutes when scored ≥40 on a 100-mm visual analog scale (VAS), and for 30 minutes on the next 2 days. There was no EA in the waiting list group (n=13). OUTCOME MEASURES: Primary outcomes were menstrual pain, resistance index (RI), and the ratio between peak systolic to end-diastolic flow velocity (S/D) in uterine arteries. Secondary outcomes included verbal rating scale (VRS) and retrospective symptom scale (RSS). RESULTS: The SP6 group had a highly significant reduction in VAS scores compared with the waiting list group (-23.19mm, 95% confidence interval [CI]-32.06 to -14.33, P<0.0001), GB39 group (-18.58mm, 95% CI -27.29 to -9.88, P<0.0001) and the non-meridian point group (-20.78mm, 95% CI -29.82 to -11.73, P<0.0001), respectively. A significant reduction in VRS scores was found in the SP6 group compared with the GB39 group (P=0.034) and the non-meridian point group (P=0.038). There were no significant differences of RI, S/D-values and RSS scores among the four groups (P>0.05). CONCLUSIONS: EA at SP6 can immediately relieve menstrual pain and minimize the influence of pain on daily life compared with GB39 and an adjacent non-meridian point. The data preliminarily show the specificity of SP6 for the immediate pain relief of primary dysmenorrhea.


Assuntos
Pontos de Acupuntura , Dismenorreia/terapia , Eletroacupuntura , Artéria Uterina/fisiologia , Útero/irrigação sanguínea , Adolescente , Adulto , Método Duplo-Cego , Dismenorreia/fisiopatologia , Eletroacupuntura/efeitos adversos , Feminino , Humanos , Medição da Dor , Estudos Prospectivos , Fluxo Sanguíneo Regional/fisiologia , Resultado do Tratamento , Adulto Jovem
14.
J Assist Reprod Genet ; 26(7): 415-20, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19697118

RESUMO

PURPOSE: The aim of this study is to determine whether inclusion of caspase inhibitor can improve the efficacy of cryopreservation of ovarian tissue. METHODS: Mice were randomly assigned to the Group A (fresh control group) Group B (inclusion of caspase inhibitor) and Group C (non-inclusion of caspase inhibitor). Ovarian tissue in Group B and Group C was vitrified-thawed. TUNEL assay and Bax protein detection were measured after cryopreservation. The mice in all groups received autotransplantation. The number of days before the resumption of estrous cycles was measured daily from the 5th day after surgery, and the percentage of cells expressing PCNA in grafts was measured one month following transplantation. RESULTS: The incidence of TUNEL positive follicles in Group B was significantly higher than that in Group C. Similarly, the percentage of follicles expressing Bax protein in Group B was significantly higher than that in Group C. The number of days before the resumption of estrous cycles in Group B was significantly less than that in Group C. In addition, the percentage of follicular and stromal cells expressing PCNA of grafts in Group B was significantly higher than that in Group C. CONCLUSIONS: The global caspase inhibitor Z-VAD-FMK decreases the incidence of apoptosis of ovarian tissue induced by cryopreservation, and inclusion of caspase inhibitor improves the efficacy of cryopreservation of ovarian tissue.


Assuntos
Clorometilcetonas de Aminoácidos/farmacologia , Criopreservação/métodos , Inibidores de Cisteína Proteinase/farmacologia , Ovário/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Inibidores de Caspase , Meios de Cultura , Feminino , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Endogâmicos ICR , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/metabolismo , Ovário/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Proteína X Associada a bcl-2/metabolismo
15.
Cancer Sci ; 100(6): 1051-61, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19432902

RESUMO

The regulatory mechanism of endometrial carcinoma and the signal transduction pathways involved in hormone action are poorly defined. It has become apparent that the G protein-coupled receptor (GPR) 30 mediates the non-genomic signaling of 17beta-estradiol (E2). Here we show that GPR30 is highly expressed in endometrial cancer tissues and cancer cell lines and positively regulates cell proliferation and invasion. GPR30 expression was detected in 50 human endometrial carcinomas. The transcription level of GPR30 was significantly higher in the tissue of endometrial carcinoma than in normal endometrium (P < 0.05). Immunohistochemical assays revealed that the positive expression rate of GPR30 protein in endometrial carcinoma tissue (35/50, 70%) was statistically higher than in normal endometrium tissue (8/30, 26.67%) (chi2 = 14.16, P = 0.0002). GPR30 overexpression was correlated with high-grade endometrial carcinoma. GPR30 expression was also found in two human endometrial cancer cell lines: RL95-2 (estrogen receptor positive) and KLE (estrogen receptor negative). The roles of GPR30 in proliferative and invasive responses to E2 and G1, a non-steroidal GPR30-specific agonist, in RL95-2 and KLE cell lines were then explored. We showed that E2 and G1 could initiate the MAPK/ERK mitogen-activated protein kinase pathway in both cell lines. What's more, E2 and G1 promoted KLE and RL95-2 proliferation and stimulated matrix metalloproteinase production and activity via the GPR30-mediated MEK/ERK mitogen-activated protein kinase pathway, as well as increased interleukin-6 secretion. These findings suggest that GPR30-mediated non-genomic signaling could play an important role in endometrial cancer.


Assuntos
Neoplasias do Endométrio/patologia , Estrogênios/fisiologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Interleucina-6/metabolismo , MAP Quinase Quinase 1/metabolismo , Receptores de Estrogênio/fisiologia , Receptores Acoplados a Proteínas G/fisiologia , Divisão Celular , Linhagem Celular Tumoral , Primers do DNA , Neoplasias do Endométrio/enzimologia , Endométrio/patologia , Endométrio/fisiologia , Endométrio/fisiopatologia , Ativação Enzimática , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Metaloproteinases da Matriz/metabolismo , Pessoa de Meia-Idade , Invasividade Neoplásica , Receptores Acoplados a Proteínas G/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais
16.
J Assist Reprod Genet ; 26(2-3): 137-42, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19205869

RESUMO

PURPOSE: The aim of this experiment is to detect effects of varying levels of sucrose on vitrified ovarian tissues. METHODS: Ovarian tissues of mice were vitrified-thawed. Mice were randomly assigned to the fresh control group and experimental groups. According to different concentration of sucrose in vitrification solution, the experimental groups were randomly divided into Group I (0.2 M sucrose), Group II (0.4 M sucrose), Group III (0.8 M sucrose) and Group IV (1.6 M sucrose). Cytology was followed throughout the oophorectomy and transplantation period. Hormone levels and density of follicle were measured 1 month after transplantation. RESULTS: The number of days before the resumption of estrous cycles in control group was significantly smaller than those in all of experimental groups. The serum estradiol levels of mice and the follicular density of ovarian grafts in control group were significantly higher than those in all of experimental groups. In addition, the number of days before the resumption of estrous cycles in Group II and Group III were smaller than those in Group I and Group IV. The serum estradiol levels of mice and the follicular density of ovarian grafts in Group II and Group III were significantly higher than those in Group I and Group IV. However, no difference was observed in the number of days before the resumption of estrous cycles and the serum estradiol levels between Group II and Group III. A similar follicular density was also observed in Group II and Group III. CONCLUSION: Sucrose concentration of 0.4 M or 0.8 M in cryoprotective media is suitable for vitrifying mouse ovarian tissues.


Assuntos
Criopreservação/métodos , Crioprotetores , Ovário/transplante , Sacarose , Animais , Estradiol/sangue , Ciclo Estral/fisiologia , Feminino , Camundongos , Folículo Ovariano/fisiologia , Técnicas de Reprodução Assistida
17.
Luminescence ; 22(5): 393-400, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17471452

RESUMO

The interaction between surfactant and fluorescein was studied, using a fluorescence spectroscopy and flow-injection (FI) chemiluminescence (CL) method. It was found that the cationic surfactant cetyltrimethylammonium bromide (CTAB) could cause the structural transformation of fluorescein from the quinone to the spirolactone form, and greatly enhance the CL intensity of the fluorescein-human serum albumin (HSA) complex. Based on this finding, a rapid and sensitive FI-CL method was developed for the determination of HSA. Under the optimum conditions, the proposed method has a linear range of 0.05-24.0 microg/mL, with a detection limit of 0.03 microg/mL for HSA (3sigma). The relative standard deviation (RSD) of 1.2 microg/mL HSA (n = 8) is 0.8%. The method was applied to the determination of protein content in urine samples, with satisfactory results. Density functional theory was used to study the mechanism of surfactant-enhanced CL intensity of the fluorescein-HSA complex.


Assuntos
Compostos de Cetrimônio/química , Fluoresceína/química , Medições Luminescentes/métodos , Albumina Sérica/análise , Hipoclorito de Sódio/química , Tensoativos/química , Cátions , Cetrimônio , Análise de Injeção de Fluxo/instrumentação , Análise de Injeção de Fluxo/métodos , Humanos , Concentração de Íons de Hidrogênio , Cinética , Medições Luminescentes/instrumentação , Estrutura Molecular , Sensibilidade e Especificidade
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