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1.
Endocrinology ; 163(8)2022 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-35731831

RESUMO

CONTEXT: Spontaneous abortion (SA) is a common disorder in early pregnancy. Circular RNAs (circRNAs) have been reported to exert important regulatory effects on trophoblast function and embryo development. OBJECTIVE: The aim of this study was to explore whether and how circRNAs regulate trophoblast function in SA during early pregnancy. METHODS: Cell proliferation, 5-bromo-2-deoxyuridine (BrdU) staining, Transwell, immunofluorescence, Western blot, RNA pull-down, and dual luciferase reporter assays were performed to investigate the effect of circRNA cyclin B1 (circ-CCNB1) on trophoblast function in HTR-8/SVneo and JEG-3 cells. RESULTS: An in vitro study demonstrated that upregulation of circ-CCNB1 significantly inhibited trophoblast proliferation and invasion compared with the controls using HTR-8/SVneo and JEG-3 cells, respectively. Moreover, miR-223 was downregulated in the villous tissues of patients with SA and was further predicted and shown to negatively interact with circ-CCNB1, which is involved in trophoblast proliferation and invasion. Using bioinformatics tools and subsequent RNA pull-down and dual luciferase assays, we found that miR-223 directly targets seven in absentia homolog-1 (SIAH1) and that upregulation of miR-223 decreased circ-CCNB1-induced SIAH1 expression levels in HTR-8/SVneo cells. Interestingly, upregulation of circ-CCNB1 suppressed trophoblast proliferation and invasion through inhibition of CCNB1 nuclear translocation induced by SIAH1. Downregulation of SIAH1 enhanced circ-CCNB1-suppressed CCNB1 nuclear protein expression in trophoblast cells. CONCLUSION: Circ-CCNB1 served as a modulator of trophoblast proliferation and invasion by sponging miR-223, thus forming a regulatory network of circ-CCNB1/miR-223/SIAH1 in modulating CCNB1 nuclear translocation, which enabled us to elucidate the molecular mechanisms involved in normal embryo implantation or in SA.


Assuntos
Aborto Espontâneo , MicroRNAs , Aborto Espontâneo/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Ciclina B1/genética , Ciclina B1/metabolismo , Feminino , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Gravidez , RNA Circular/genética , Trofoblastos/metabolismo
2.
J Zhejiang Univ Sci B ; 21(6): 485-494, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32478494

RESUMO

Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (ΔΨm), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P<0.05), and RSV did not significantly increase the sperm motility (0.44 vs. 0.40, P>0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ΔΨm. RSV treatment could increase the rate of high mitochondrial ΔΨm of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 µmol/L vs. 0.25 µmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor- and mitochondria-mediated apoptotic pathways.


Assuntos
Apoptose/efeitos dos fármacos , Resveratrol/farmacologia , Preservação do Sêmen/veterinária , Espermatozoides/efeitos dos fármacos , Acrossomo/efeitos dos fármacos , Animais , Masculino , Potencial da Membrana Mitocondrial , Espécies Reativas de Oxigênio/metabolismo , Motilidade dos Espermatozoides/efeitos dos fármacos , Suínos
3.
Biomed Pharmacother ; 117: 109092, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31203134

RESUMO

BACKGROUND: 17ß-Estradiol (E2) is a critical regulator of trophoblast function during pregnancy. Serum- and glucocorticoid-inducible kinase (SGK1) has been shown to regulate specific cellular targets downstream of E2. However, whether and how SGK1 directly mediates the regulatory effects of E2 on trophoblasts functions remain unknown. METHODS: SGK1 expression in human villous samples and serum E2 levels were measured in women with early pregnancy loss (EPL) and healthy pregnant women. The effect of E2 on SGK1 regulation was assessed using luciferase reporter gene assay and Chromatin Immunoprecipitation assay. The mediation of regulatory effects of E2 by SGK1 on trophoblast functions including cell viability, invasion and related signaling molecules such as B cell leukemia/lymphoma 6, E-cadherin, matrix metalloproteinase 2, α-ENaC, vascular endothelial growth factor, and the phosphorylation status of FOXO1 and AKT were evaluated in HTR8/SVneo cells transfected with SGK1 knockdown plasmid with/without E2 treatment. RESULTS: SGK1 protein levels in human villous samples and serum E2 levels were decreased in patients with EPL compared to controls. E2 (10 nM) increased SGK1 promoter activity directly through estrogen receptor. E2-activated SGK1 enhanced cell viability, invasion and downstream targets in trophoblast cells. SGK1 knockdown abrogated the above responses to E2 treatment. CONCLUSIONS: SGK1 mediates the effects of E2 on trophoblast viability and invasion, suggesting that SGK1 acts as a key node in regulating the cross-talk at the feto-maternal interface during the development of placenta and might be a potential therapeutic target for EPL.


Assuntos
Sobrevivência Celular/fisiologia , Estradiol/metabolismo , Proteínas Imediatamente Precoces/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Trofoblastos/metabolismo , Caderinas/metabolismo , Linhagem Celular , Movimento Celular/fisiologia , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Placenta/metabolismo , Gravidez , Regiões Promotoras Genéticas/fisiologia , Receptores de Estrogênio/metabolismo
4.
Medicine (Baltimore) ; 94(13): e636, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25837753

RESUMO

Langerhans cell sarcoma (LCS) is a rare tumor with markedly malignant cytological features originating from Langerhans cells. LCS diagnosis is difficult and requires differentiation from other malignant tumors and Langerhans cell histiocytosis (LCH). Immunochemical antibodies, such as langerin, S-100 protein, and CD1a, have been used to diagnose LCS, but the results are crossed with LCH. To determine more significant biomarkers of LCS, we studied the expression and distribution pattern of Wilms tumor 1 (WT1) and cluster of differentiation 44 (CD44) in LCS. A broad panel of antibodies was used for immunohistochemical technology. Simultaneously, dual immunofluorescence staining examination and fluorescence in situ hybridization staining methods were used to study the location of WT1 and CD44 in LCS tumor cells. The results showed that tumor cells expressed WT1, CD44, and other special Langerhans cell markers (langerin, CD1a, and S-100 protein). LCS cells in all the cases showed normal cytogenetic findings without overexpression of WT1 and CD44. The expression of WT1 and CD44 was observed on langerin tumor cells by dual immunofluorescence staining examination in LCS. Our results suggest that WT1 and CD44 are potential biomarkers for LCS diagnosis. Clear understanding of their functional roles may further explain the pathogenesis of this highly malignant tumor and develop some novel immunotherapy strategies.


Assuntos
Receptores de Hialuronatos/sangue , Sarcoma de Células de Langerhans/sangue , Sarcoma de Células de Langerhans/diagnóstico , Tumor de Wilms/sangue , Adulto , Antígenos CD/sangue , Antígenos CD1/sangue , Biomarcadores Tumorais , Diagnóstico Diferencial , Feminino , Humanos , Hibridização in Situ Fluorescente , Sarcoma de Células de Langerhans/patologia , Lectinas Tipo C/sangue , Masculino , Lectinas de Ligação a Manose/sangue , Pessoa de Meia-Idade , Proteínas S100/sangue
5.
Turk J Gastroenterol ; 25 Suppl 1: 147-52, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25910294

RESUMO

BACKGROUND/AIMS: The only hope for a cure from hepatocellular carcinoma (HCC) rests on early diagnosis. The present study aims to determine serum peptidome patterns for early diagnosis of HCC. MATERIALS AND METHODS: To identify novel peptidome patterns for diagnosing HCC, serum from31 healthy volunteers and 32 HCC patients were subjected to a comparative proteomic analysis using a ClinProt Kit combined with mass spectrometry (MS). This approach allows the determination of peptidome patterns that are able to differentiate the HCC from healthy volunteers. For further validation, the diagnostic and differential diagnostic capabilities of the peptidome patterns were verified blindly by an independent group of sera consisted of 31 HCC, 23 liver fibrosis and 33 healthy volunteers. RESULTS: A Quick Classifier Algorithm was used to construct the peptidome patterns for the identification of HCC from the control samples. One of the identified peaks at m/z 7771 was used to construct the peptidome patterns with almost 100% accuracy. Furthermore, the peptidome patterns could also differentiate the validation group with high accuracy. CONCLUSION: These results suggest that the ClinProt Kit combined with MS achieves significantly high accuracy for HCC diagnosis and differential diagnosis.


Assuntos
Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/diagnóstico , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/diagnóstico , Peptídeos/sangue , Diagnóstico Diferencial , Detecção Precoce de Câncer/métodos , Feminino , Humanos , Fenômenos Magnéticos , Masculino , Espectrometria de Massas/métodos , Microesferas , Pessoa de Meia-Idade , Proteômica
6.
Mol Biol Rep ; 39(9): 9179-86, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22733488

RESUMO

Early pregnancy loss (EPL) is one of the most common complications of human reproduction. Combined with our previous proteomic studies on villous and decidual tissues of EPL, we found that alterations of the proteins involved in oxidative stress (OS), unfolded protein response (UPR) and proteolysis presented a complex and dynamic interaction at the maternal-fetal interface. In the present study, we developed a cell model of OS using normal decidual cells to examine cell viability and expression levels of proteins related to endoplasmic reticulum stress (ER stress) and UPR. We found that glucose regulated protein 78 (GRP 78) and ubiquitinated proteins were significantly up-regulated in hydrogen peroxide (H(2)O(2)) treated decidual cells in a dose-dependent manner. Excessive OS could influence proper function of UPR by decreasing VCP in decidual cells, thereby leading to cell damage as well as inhibition of cell growth and activation of apoptosis. Furthermore, when pretreated with MG 132, a pharmacological inhibition of the proteasome, the H(2)O(2) treated decidual cells became less viable and could not up-regulate the expression level of GRP 78 to resolve the protein-folding defects, which indicating that malfunction of UPR in decidual cells might aggravate the inhibitory effect of OS in decidual cells. The present results reveal that abnormal protein profiles associated with OS induced ER stress and malfunction of UPR might be involved in the development of EPL, and OS and ER stress are potential targets for pregnant care and prognosis in normal pregnancy and its disorders.


Assuntos
Aborto Espontâneo/etiologia , Decídua/metabolismo , Estresse do Retículo Endoplasmático , Estresse Oxidativo , Adenosina Trifosfatases/metabolismo , Proteínas de Ciclo Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Decídua/citologia , Chaperona BiP do Retículo Endoplasmático , Feminino , Proteínas de Choque Térmico/metabolismo , Humanos , Peróxido de Hidrogênio/farmacologia , Gravidez , Ubiquitina/metabolismo , Resposta a Proteínas não Dobradas , Proteína com Valosina
7.
BMC Med ; 10: 26, 2012 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-22413869

RESUMO

BACKGROUND: Early pregnancy loss (EPL) is a frustrating clinical problem, whose mechanisms are not completely understood. DNA methylation, which includes maintenance methylation and de novo methylation directed by DNA methyltransferases (DNMTs), is important for embryo development. Abnormal function of these DNMTs may have serious consequences for embryonic development. METHODS: To evaluate the possible involvement of DNA methylation in human EPL, the expression of DNMT proteins and global methylation of DNA were assessed in villous or decidua from EPL patients. The association of maintenance methylation with embryo implantation and development was also examined. RESULTS: We found that DNMT1 and DNMT3A were both expressed in normal human villous and decidua. DNMT1 expression and DNA global methylation levels were significantly down-regulated in villous of EPL. DNMT3A expression was not significantly changed in the EPL group compared to controls in either villous or decidua. We also found that disturbance of maintenance methylation with a DNMT1 inhibitor may result in a decreased global DNA methylation level and impaired embryonic development in the mouse model, and inhibit in vitro embryo attachment to endometrial cells. CONCLUSIONS: Our results demonstrate that defects in DNA maintenance methylation in the embryo, not in the mother, are associated with abnormal embryonic implantation and development. The findings of the current study provide new insights into the etiology of EPL.


Assuntos
Metilação de DNA , Metilases de Modificação do DNA/metabolismo , Desenvolvimento Embrionário/fisiologia , Aborto Espontâneo/etiologia , Animais , Decídua/enzimologia , Modelos Animais de Doenças , Feminino , Feto/enzimologia , Humanos , Camundongos , Camundongos Endogâmicos ICR , Gravidez
8.
J Clin Endocrinol Metab ; 96(3): E493-7, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21177787

RESUMO

BACKGROUND: Oxidative stress is a common pathological background for different etiologies of early pregnancy loss (EPL). It has been suggested that elevated reactive oxygen species trigger endoplasmic reticulum (ER) stress by influencing ER function. However, it is unclear whether ER stress is associated with EPL. OBJECTIVES: The aim of the study was to determine whether and how ER stress occurs during the development of EPL. APPROACHES: Proteomic analysis was performed on decidua from women with EPL, and then ER stress markers, redox status, apoptotic features, and cell viability were analyzed in EPL decidual cells (DCs). RESULTS: EPL decidua were characterized by decreased levels of glucose-regulated protein 78 (GPR78) and valosin-containing protein and burdened with ubiquitinated proteins. Evidence of ER stress-induced apoptosis in EPL DCs was demonstrated by extensive dilation of ER, morphological features of apoptosis, and activation of caspase-4 and caspase-12. Furthermore, H(2)O(2) reduced the viabilities in both EPL and control DCs, whereas EPL DCs were more vulnerable to additional OS challenge than the controls as a result of failed induction of GRP78 expression. The cell survival percentages of DCs were dose-dependently reduced by H(2)O(2) and could be reversed in the presence of vitamin E. This effect was partly mediated by reducing the amount of misfolded proteins rather than regulating GRP78 expression. CONCLUSIONS: The sum of these observations demonstrate for the first time that sustained ER stress occurs in EPL DCs and the potentially vicious relationship between ER stress and oxidative stress is likely to play an important role in the development of EPL.


Assuntos
Aborto Espontâneo/metabolismo , Aborto Espontâneo/patologia , Decídua/citologia , Retículo Endoplasmático/metabolismo , Estresse Oxidativo/fisiologia , Aborto Espontâneo/genética , Adenosina Trifosfatases/metabolismo , Adulto , Western Blotting , Caspases/metabolismo , Proteínas de Ciclo Celular/metabolismo , Separação Celular , Sobrevivência Celular/fisiologia , Relação Dose-Resposta a Droga , Chaperona BiP do Retículo Endoplasmático , Feminino , Proteínas de Choque Térmico/genética , Humanos , Peróxido de Hidrogênio/farmacologia , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Gravidez , Proteômica , Espécies Reativas de Oxigênio/metabolismo , Proteína com Valosina
9.
Huan Jing Ke Xue ; 31(10): 2525-31, 2010 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-21229772

RESUMO

By applying bacteria as anodic catalyst, microbial fuel cell (MFC) can directly convert biomass energy into electrical energy, provided a new way for biomass utilization. Previous studies showed that the substrates and their concentration substantially affected performance of MFC. High power output was obtained when simple organic such as volatile fatty acids (VFA), alcohols or glucose was used as substrate. However, physical, chemical or even biological pretreatment methods were needed when substrate was complex organic. Addition of simple organic as co-substrate was also demonstrated to be an efficient way for refractory compounds degradation in MFC. Using biomass as substrates, MFC will be applied in area such as bioenergy recovery from wastewater, power supply in outfield and biosensors.


Assuntos
Fontes de Energia Bioelétrica/tendências , Conservação de Recursos Energéticos/métodos , Eletricidade , Eliminação de Resíduos Líquidos/métodos , Especificidade por Substrato
10.
Huan Jing Ke Xue ; 31(11): 2804-10, 2010 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-21250469

RESUMO

Facultative tube methods were adopted for isolation from biofilm in two compartments of a baffled air-cathode microbial fuel cell (BAFMFC). The separated strains were then putted into a sterile cubic MFC individually under the same condition in order to evaluate its electrogenic characteristic. Electrochemical impedance spectrometer (EIS) was employed for the internal resistance testing. It can be found that ohmic internal resistance was around 25omega, indicating the cell performance determined only by activity of the strain. Nineteen strains were obtained, seven of which had voltages over 200 mV under 1 000omega. The strain (A2), owned the highest electrogenic activity in compartment A, has a voltage of 328 mV with the maximum power density 165.1 mW/m2. The maximum power density of 214.4 mW/m2 was produced by the isolate B1 (the best exoelectrogenesis in compartment B), while the maximal voltage of 241 mV was achieved. Identification of fatty acids reveals that A2 belongs to Enterobacteriaceae, and B1 belongs to the genus Bacillus.


Assuntos
Bacillus/isolamento & purificação , Bacillus/metabolismo , Fontes de Energia Bioelétrica/microbiologia , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/metabolismo , Eletricidade , Eletrodos
11.
Zhonghua Liu Xing Bing Xue Za Zhi ; 28(7): 704-7, 2007 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-18069565

RESUMO

OBJECTIVE: To study the clinical effect under highly active anti-retroviral therapy (HAART) in AIDS patients and for improving the curative effect and prognosis. METHODS: Epidemiological method was used from five aspects to describe the post-treatment clinical symptoms of 181 AIDS patients in Suizhou, and to evaluate the change of virus load and immune function of 79 AIDS patients. Data was doubly recorded by Epi Data and database was set up by SPSS 13.0 for analysis. RESULTS: The effective powers of anomal-fever, cough, diarrhoea, lymphadenectasis, weight drop, erythra, mycotic infection were 81.39%, 85.00%, 84.62%, 81.89%, 82.86%, 66.07% and 45.45% respectively. CD4+ T lymphocyte count rose obviously after treatment, with an averag of 276 x 10(6) cells/ml (65 x 10(6)-824 x 10(6) cells/ml), an 129 x 10(6) cells/ml increase in three months and was 294 x 10(6) cells/ml (102 x 10(6)-750 x 10(6) cells/ml) in six months. The count change of CD4+ T lymphocyte between 3 months and 6 months did not show sigificant difference. The number of deaths among drug withdrawals was 14, with a case fatality rate as 29.79%; while the number of deaths among non-drug withdrawals was 3, with the case fatality rate as 2.24%. CONCLUSION: Results through this study showed that HAART could obviously improve the clinical symptom of AIDS patients, and to increase the number of virus load. Improving the compliance could also reduce the case fatality rate.


Assuntos
Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Terapia Antirretroviral de Alta Atividade/métodos , Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/patologia , Síndrome da Imunodeficiência Adquirida/virologia , Adulto , Linfócitos T CD4-Positivos/efeitos dos fármacos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Inquéritos e Questionários , Resultado do Tratamento
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