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1.
Int J Biol Sci ; 19(3): 994-1006, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36778116

RESUMO

Cardiac fibrosis is a common pathological cardiac remodeling in a variety of heart diseases, characterized by the activation of cardiac fibroblasts. Our previous study uncovered that promyelocytic leukemia protein (PML)-associated SUMO processes is a new regulator of cardiac hypertrophy and heart failure. The present study aimed to explore the role of PML in cardiac fibroblasts activation. Here we found that PML is significantly upregulated in cardiac fibrotic tissue and activated cardiac fibroblasts treated with transforming growth factor-ß1 (TGF-ß1). Gain- and loss-of-function experiments showed that PML impacted cardiac fibroblasts activation after TGF-ß1 treatment. Further study demonstrated that p53 acts as the transcriptional regulator of PML, and participated in TGF-ß1 induced the increase of PML expression and PML nuclear bodies (PML-NBs) formation. Knockdown or pharmacological inhibition of p53 produced inhibitory effects on the activation of cardiac fibroblasts. We further found that PML also may stabilize p53 through inhibiting its ubiquitin-mediated proteasomal degradation in cardiac fibroblasts. Collectively, this study suggests that PML crosstalk with p53 regulates cardiac fibroblasts activation, which provides a novel therapeutic strategy for cardiac fibrosis.


Assuntos
Proteína da Leucemia Promielocítica , Fator de Crescimento Transformador beta1 , Proteína Supressora de Tumor p53 , Humanos , Fibroblastos/metabolismo , Fibrose , Coração , Fator de Crescimento Transformador beta1/farmacologia , Proteína Supressora de Tumor p53/metabolismo , Proteína da Leucemia Promielocítica/metabolismo
2.
BMJ Open ; 12(10): e063803, 2022 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-36198457

RESUMO

BACKGROUND: As one of the most common stroke sequelae, poststroke cognitive impairment significantly impacts 17.6%-83% of survivors, affecting their rehabilitation, daily living and quality of life. Improving cognitive abilities among patients in stroke recovery is therefore critical and urgent. Transcutaneous auricular vagus nerve stimulation (TAVNS) is a non-invasive, safe, cost-effective treatment with great potential for improving the cognitive function of poststroke patients. This clinical research will evaluate the effectiveness, and help elucidate the possible underlying mechanisms, of TAVNS for improving poststroke cognitive function. METHODS AND ANALYSIS: A single-centre, parallel-group, allocation concealment, assessor-blinded randomised controlled clinical trial. We will allocate 88 recruited participants to the TAVNS or sham group for an intervention that will run for 8 weeks, 5 days per week with twice daily sessions lasting 30 min each. Blood tests will be performed and questionnaires issued at baseline and 8-week and 12 week follow-ups. Primary outcomes will be changes in cognitive function scores. Secondary outcomes will be changes in activities of daily living, quality of life and serum oxidative stress indicators. ETHICS AND DISSEMINATION: The Ethics Committee of the First Affiliated Hospital of Hunan University of Chinese Medicine has approved the protocol (No. HN-LL-YJSLW-2022200). Findings will be published in peer-reviewed academic journals and presented at scientific conferences. TRIAL REGISTRATION NUMBER: ChiCTR2200057808.


Assuntos
Disfunção Cognitiva , Acidente Vascular Cerebral , Estimulação do Nervo Vago , Atividades Cotidianas , Disfunção Cognitiva/etiologia , Disfunção Cognitiva/terapia , Humanos , Qualidade de Vida , Ensaios Clínicos Controlados Aleatórios como Assunto , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/terapia , Estimulação do Nervo Vago/métodos
4.
Acta Pharmacol Sin ; 42(7): 1139-1149, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33318625

RESUMO

This study aimed to investigate the inhibitory effect of EM-2, a natural active monomer purified from Elephantopusmollis H.B.K., on the proliferation of human hepatocellular carcinoma cells and the molecular mechanism involved. The results from the MTT assay revealed that EM-2 significantly inhibited the proliferation of human hepatocellular carcinoma (HCC) cells in a dose-dependent manner but exhibited less cytotoxicity to the normal liver epithelial cell line LO2. EdU staining and colony formation assays further confirmed the inhibitory effect of EM-2 on the proliferation of Huh-7 hepatocellular carcinoma cells. According to the RNA sequencing and KEGG enrichment analysis results, EM-2 markedly activated the MAPK pathway in Huh-7 cells, and the results of Western blotting further indicated that EM-2 could activate the ERK and JNK pathways. Meanwhile, EM-2 induced apoptosis in a dose-dependent manner and G2/M phase arrest in Huh-7 cells, which could be partially reversed when treated with SP600125, a JNK inhibitor. Further study indicated that EM-2 induced endoplasmic reticulum stress and blocked autophagic flux in Huh-7 cells by inhibiting autophagy-induced lysosome maturation. Inhibition of autophagy by bafilomycin A1 could reduce cell viability and increase the sensitivity of Huh-7 cells to EM-2. In conclusion, our findings revealed that EM-2 not only promoted G2/M phase arrest and activated ER stress but also induced apoptosis by activating the JNK pathway and blocked autophagic flux by inhibiting autolysosome maturation in Huh-7 hepatocellular carcinoma cells. Therefore, EM-2 is a potential therapeutic drug with promising antitumor effects against hepatocellular carcinoma and fewer side effects.


Assuntos
Antineoplásicos/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Lactonas/farmacologia , Neoplasias Hepáticas/tratamento farmacológico , Sesquiterpenos/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Humanos , Lisossomos/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos
6.
Cardiovasc Ultrasound ; 17(1): 14, 2019 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-31325956

RESUMO

BACKGROUND: Postnatal outcome of fetuses with isolated single umbilical artery (SUA) is determined mainly by right ventricular function. Our study examined whether there are differences in right ventricular function during the gestation period of fetuses with isolated SUA compared to healthy fetuses. METHODS: A prospective study was conducted on 77 fetuses with isolated SUA and 77 gestational age-matched controls from 22 to 39 weeks. For gestational age grouping, the isolated SUA fetuses and the control fetuses were divided into the second trimester group (22-27 weeks; 43 fetuses) and third trimester group (28-39 weeks; 34 fetuses). The fetal tricuspid annular plane systolic excursion (f-TAPSE) M-mode was applied to the tricuspid annulus, parallel to the ventricular septum, and the amplitude of the resulting wave was assessed using spatiotemporal image correlation (STIC) M-mode. We investigated the possible changes to the STIC M-mode indices during the course of pregnancy in both the isolated SUA and control groups. The relationship between f-TAPSE and gestational age was analyzed. Additionally, the correlations between f-TAPSE and birth weight was analyzed, and the birth weight differences between the isolated SUA and control groups in the third trimester were analyzed according to postpartum results. RESULTS: There was a significant difference in f-TAPSE between isolated SUA and control group in the third trimester (P < 0.05). There were significant correlations between gestational age (GA) and f-TAPSE among control fetuses (R2 = 0.9049; P < 0.01). A significant, positive correlation between GA and f-TAPSE was also found with isolated SUA fetuses (R2 = 0.8108; P < 0.01). The prevalence of small-for-gestational-age (SGA) fetuses and of discordant birth weight fetuses was significantly higher in the isolated SUA group than in the control group. In univariate analysis, the presence of an isolated SUA was associated with lower birth weight (2940 g compared with 3260 g) and with higher prevalence of SGA (13.0% compared with 3.9%; P < 0.01). The correlations between the birth weight and f-TAPSE in the two groups were analyzed in the third trimester, and the correlation in the isolated SUA group was better than that of the control group (R2 was 0.623 and 0.463 in the isolated SUA group and the control group, respectively). CONCLUSIONS: Right ventricular function in isolated SUA is altered as early as in fetal third trimester. STIC M-mode can measure the right heart function of the fetus and may predict isolated SUA with SGA.


Assuntos
Ecocardiografia Doppler em Cores/métodos , Coração Fetal/diagnóstico por imagem , Ventrículos do Coração/fisiopatologia , Artéria Umbilical Única/diagnóstico , Ultrassonografia Pré-Natal/métodos , Artérias Umbilicais/diagnóstico por imagem , Função Ventricular Direita/fisiologia , Adulto , Feminino , Coração Fetal/fisiopatologia , Idade Gestacional , Ventrículos do Coração/diagnóstico por imagem , Ventrículos do Coração/embriologia , Humanos , Gravidez , Terceiro Trimestre da Gravidez , Estudos Prospectivos , Reprodutibilidade dos Testes , Artéria Umbilical Única/fisiopatologia , Artérias Umbilicais/anormalidades , Artérias Umbilicais/embriologia
7.
J Cancer Res Ther ; 14(Supplement): S1035-S1040, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30539842

RESUMO

AIM: Norcantharidin (NCTD) has been used as a clinical antineoplastic drug in China for several years, and diamminedichloroplatinum is a valuable clinical cancer chemotherapy agent. Here, we tried to investigate the effects of NCTD plus diamminedichloroplatinum on hepatic carcinoma in murine. MATERIALS AND METHODS: In vivo and in vitro investigations on anticancer effects of the two drugs were individually made. RESULT: In vitro, the combination of the two drugs resulted in apparent apoptosis and cell proliferation inhibitions of H22 cancer cells. Meanwhile, their coadministration in vivo produced significant suppressions of tumor growth and cancerometastasis. Further, CD31 immunohistochemistry and matrigel tube formation assay demonstrated that angiogenesis was inhibited by NCTD plus diamminedichloroplatinum in vivo and in vitro, respectively. CONCLUSION: Based on the findings, we concluded that NCTD plus diamminedichloroplatinum may have an additive anticancer efficacy because the two drugs work in different ways, and thus, their combination had inhibited cancer cell proliferations and tumor angiogenesis more effectively than either of the compounds alone.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Cisplatino/farmacologia , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Apoptose/efeitos dos fármacos , Compostos Bicíclicos Heterocíclicos com Pontes/uso terapêutico , Carcinoma Hepatocelular/secundário , Linhagem Celular Tumoral/transplante , Proliferação de Células/efeitos dos fármacos , Cisplatino/uso terapêutico , Modelos Animais de Doenças , Sinergismo Farmacológico , Feminino , Humanos , Neoplasias Hepáticas/patologia , Neoplasias Pulmonares/secundário , Camundongos , Camundongos Endogâmicos BALB C , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/patologia , Resultado do Tratamento
8.
Cancer Biother Radiopharm ; 33(1): 25-31, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29466034

RESUMO

Cancer vaccines mostly aim to induce cytotoxic T lymphocytes (CTLs) against tumors. An appropriate adjuvant is of fundamental importance for inducing cellular immune response. Since the antigen in particulate form is substantially more immunogenic than soluble form antigen, it is beneficial to interact with antigen-presenting cells membrane to induce robust CD8+ T cell activation following vaccination. Based on previous research, we designed an adjuvant formulation by combining Astragalus saponins, cholesterol, and liposome to incorporate antigen into a particulate delivery system, so as to enhance cellular immune response. Meanwhile, angiogenesis contributes to tumor growth and metastasis, and basic fibroblast growth factor (bFGF) is involved in tumor angiogenesis. Therefore, using lipo-saponins adjuvant formulation and a human recombinant bFGF antigen protein, we tried to induce bFGF-specific CTL response to inhibit tumor angiogenesis to achieve antitumor activity. After five immunizations, the lipo-saponins/bFGF complex elicited robust antibody response and markedly higher amount of interferon-γ in BALB/c mice, resulting in superior antitumor activities. Decreased microvessel density in CD31 immunohistochemistry and the lysis of vascular endothelial cells by the T lymphocytes from the immunized mice indicated that the immunity inhibited the angiogenesis of tumors and further led to the inhibition of tumors. Our data suggest that the approach to construct adjuvant formulation between liposome and Astragalus saponins appeared highly desirable, and that Astragalus saponins may be utilized as a valuable additive for enhancing the effectiveness of vaccines and stimulating an appropriate immune response that can benefit tumor therapy.


Assuntos
Adjuvantes Imunológicos/química , Vacinas Anticâncer/química , Lipossomos/química , Saponinas/química , Saponinas/imunologia , Sequência de Aminoácidos , Animais , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C
9.
Cancer Biol Ther ; 19(6): 518-524, 2018 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-29405828

RESUMO

Tumor growth and metastasis are closely related to angiogenesis. Basic fibroblast growth factor(bFGF) is an angiogenic factor, and up-regulated expression of bFGF plays a crucial role in the development and metastasis of melanoma. Therefore, in this study, we sought to achieve antitumor activity by immunity targeting bFGF which would inhibit tumor angiogenesis and simultaneously induce bFGF specific cytotoxic T lymphocytes to kill melanoma cells. A human bFGF protein was used as exogenous antigen, coupled with a saponin-liposome adjuvant formulation to enhance CTL response. The results showed that the immunity induced strong immune response and produced prominent anti-cancer activities. CD31 immunohistochemistry and alginate-encapsulated tumor cell assay displayed that tumor angiogenesis was effectively inhibited. Further, the higher production of IFN-γ and cytotoxic T lymphocyte killing assay suggested that the anti-cancer activities may mainly depend on cellular immune response, which could cause the inhibition of tumor angiogenesis and specific killing of tumor cells by bFGF-specific cytotoxic T lymphocytes. We concluded that immunotherapy targeting bFGF may be a prominent strategy for melanoma, and that the adjuvant formulation of saponin-liposome is very desirable in enhancing cytotoxic T lymphocytes response.


Assuntos
Vacinas Anticâncer/uso terapêutico , Fator 2 de Crescimento de Fibroblastos/imunologia , Imunoterapia/métodos , Melanoma/tratamento farmacológico , Animais , Vacinas Anticâncer/imunologia , Vacinas Anticâncer/farmacologia , Modelos Animais de Doenças , Feminino , Humanos , Melanoma/patologia , Camundongos
10.
Exp Mol Med ; 49(7): e357, 2017 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-28729638

RESUMO

Our study aims to investigate the roles that microRNA-214 (miR-214) plays in the epithelial mesenchymal transition (EMT) process and the development of interstitial cystitis (IC) in postmenopausal women by targeting Mitofusin 2 (Mfn2). IC bladder tissues and adjacent normal bladder tissues were collected from postmenopausal women. Immunohistochemistry (IHC) staining was conducted. The target relationship between miR-214 and Mfn2 was determined by a dual luciferase reporter gene assay. Adipose-derived mesenchymal stem cells (ADMSCs) were extracted from postmenopausal rats and assigned to the blank, mimics, miR-214 inhibitors, mimics negative control (NC), inhibitors NC, Mfn2 siRNA, miR-214 inhibitors and Mfn2 siRNA groups. Exosomes secreted by transfected ADMSCs were instilled into the bladders of postmenopausal rats. The expression of miR-214 and Mfn2 mRNA and EMT markers was assessed by qRT-PCR and western blotting. It was confirmed that Mfn2 was the target gene of miR-214 in IC. Compared with the normal bladder tissues, miR-214 decreased, but Mfn2 increased in IC bladder tissues. Compared with the blank group, the expression of miR-214 and the expression levels of N-cadherin, Fibronectin, Twist1, Snail and Vimentin mRNA and protein increased, whereas the expression levels of Mfn2, E-cadherin and ZO-1 mRNA and protein decreased in the miR-214 mimics and Mfn2 groups. The expression of MiR-214 and the expression levels of N-cadherin, Fibronectin, Twist1, Snail and Vimentin mRNA and protein decreased, whereas the expression levels of Mfn2, E-cadherin and ZO-1 mRNA and protein increased in the miR-214 inhibitors group. Our findings indicate that the inhibition of miR-214 promotes the EMT process and contributes to bladder wall fibrosis by up-regulating Mfn2, thus leading to the occurrence of IC in postmenopausal women.


Assuntos
Cistite Intersticial/metabolismo , Transição Epitelial-Mesenquimal , GTP Fosfo-Hidrolases/metabolismo , MicroRNAs/fisiologia , Proteínas Mitocondriais/metabolismo , Pós-Menopausa/metabolismo , Bexiga Urinária/patologia , Tecido Adiposo/citologia , Animais , Biomarcadores/análise , Cistite Intersticial/patologia , Modelos Animais de Doenças , Exossomos/metabolismo , Feminino , Fibrose , GTP Fosfo-Hidrolases/genética , Humanos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/antagonistas & inibidores , Proteínas Mitocondriais/genética , Inibidores da Síntese de Ácido Nucleico/farmacologia , Ratos , Ratos Sprague-Dawley , Regulação para Cima
11.
ACS Appl Mater Interfaces ; 9(31): 26234-26241, 2017 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-28726369

RESUMO

Owing to the high charge mobility and low processing temperature, ZnO is regarded as an ideal candidate for electron transport layer (ETL) material in thin-film solar cells. For the film preparation, the presently dominated sol-gel (SG) and hydrolysis-condensation (HC) methods show great potential; however, the effect of these two methods on the performance of the resulting devices has not been investigated in the same frame. In this study, the ZnO films made through SG and HC methods were applied in perovskite solar cells (Pero-SCs), and the performances of corresponding devices were compared under parallel conditions. We found that the surface morphologies and the conductivities of the films prepared by SG and HC methods showed great differences. The HC-ZnO films with higher conductivity led to relatively higher device performance, and the best power conversion efficiencie (PCE) of 12.9% was obtained; meanwhile, for Pero-SCs based on SG-ZnO, the best PCE achieved was 10.9%. The better device performance of Pero-SCs based on HC-ZnO should be attributed to the better charge extraction and transportation ability of HC-ZnO film. Moreover, to further enhance the performance of Pero-SCs, a thin layer of pristine C60 was introduced between HC-ZnO and perovskite layers. By doing so, the quality of perovskite films was improved, and the PCE was elevated to 14.1%. The preparation of HC-ZnO film involves relatively lower-temperature (maximum 100 °C) processing; the films showed better charge extraction and transportation properties and can be a more promising ETL material in Pero-SCs.

12.
Eur Spine J ; 25(6): 1945-52, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-25047654

RESUMO

PURPOSE: This prospective study analyzes clinical characteristics and outcomes of sacral extradural spinal meningeal cysts (SESMC) without spinal nerve root fibers (SNRF) undergoing neck transfixion. METHODS: Using the relationship between the cysts and SNRF, SESMCs were divided into two types: cysts with SNRF known as Tarlov cysts and cysts without. If the SESMCs were identified as those without SNRFs, the neck of the cyst was transfixed, ligated and the remaining cyst wall removed distal to the clip. The improved Japanese Orthopedic Association (IJOA) scoring system was used to evaluate preoperative and postoperative neurological functions of the patients. RESULTS: Twenty-seven patients were included in this study. The average age was 42.7 ± 11.93 years. The mean preoperative IJOA score was 17.5 ± 2.47, and postoperative IJOA score was 19.1 ± 1.41. The difference between preoperative and postoperative IJOA scores was statistically significant (t = -3.75, P = 0.001), with a significant improvement in neurological function after surgery. Among the improvements in neurological function, the most significant was bowel/bladder function (z = -2.33, P = 0.02). CONCLUSION: Most patients experienced significant improvement in their neurological function after surgery. The most significant area of neurological improvement was bowel/bladder dysfunction, however, preoperative stool or urine incontinence did not recover completely.


Assuntos
Procedimentos Neurocirúrgicos , Cistos de Tarlov/cirurgia , Adolescente , Adulto , Incontinência Fecal/etiologia , Incontinência Fecal/cirurgia , Feminino , Humanos , Hipestesia/etiologia , Hipestesia/cirurgia , Masculino , Pessoa de Meia-Idade , Debilidade Muscular/etiologia , Debilidade Muscular/cirurgia , Dor/etiologia , Dor/cirurgia , Prognóstico , Estudos Prospectivos , Disfunções Sexuais Fisiológicas/etiologia , Disfunções Sexuais Fisiológicas/cirurgia , Cistos de Tarlov/complicações , Transtornos Urinários/etiologia , Transtornos Urinários/cirurgia , Adulto Jovem
13.
Artigo em Inglês | MEDLINE | ID: mdl-26557854

RESUMO

Tet is a type of alkaloid extracted from Stephania tetrandra, and it has recently been demonstrated that Tet can protect against inflammation and free radical injury and inhibit the release of inflammatory mediators. The present study was designed to observe the protective effect of Tet on sodium taurocholate-induced severe acute pancreatitis (SAP). The rat model of SAP was induced by retrograde bile duct injection of sodium taurocholate and then treated with Verapamil and Tet. The results showed that Tet can reduce NF-κB activation in pancreas issue, inhibit the SAP cascade, and improve SAP through inducing pancreas acinar cell apoptosis and stabilizing intracellular calcium in the pancreas, thus mitigating the damage to the pancreas. Our study revealed that Tet may reduce systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndromes (MODS) to protect against damage, and these roles may be mediated through the NF-κB pathway to improve the proinflammatory/anti-inflammatory imbalance.

14.
Auris Nasus Larynx ; 42(5): 353-9, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25933584

RESUMO

OBJECTIVE: The pharyngocutaneous fistula (PCF) is the troublesome complication after total laryngectomy. Despite a large number of investigations having been performed, there is still controversy about which factors are most significant for PCF. The objective of the present meta-analysis was to analyze the potential risk factors for PCF after total laryngectomy. DATA SOURCES: Published English-language literature. REVIEW METHODS: PubMed, Ovid, Cochrane, and Web of Science databases were systematically searched using multiple search terms. Twenty-one studies with 3832 patients were identified. The quality of evidence was assessed by The National Institute for Health and Clinical Excellence. RESULTS: Sixteen studies involving 2598 patients were included for the meta-analysis. The results showed that, tumor subsite (RR=0.64, 95% CI 0.47-0.88, P<0.01), T stage (RR=0.70, 95% CI 0.51-0.96, P=0.03), previous radiotherapy (RR=0.62, 95% CI 0.46-0.84, P<0.01), postoperative hemoglobin <12.5g/L (RR=0.46, 95% CI 0.27-0.76, P<0.01), and surgical margin (RR=0.41, 95% CI 0.22-0.74, P<0.01) were the risk factors associated with the development of PCF. CONCLUSIONS: From the results of our study, several significant risk factors for PCF are identified. Methodologically high-quality comparative studies are needed for further evaluation.


Assuntos
Fístula Cutânea/epidemiologia , Neoplasias Laríngeas/cirurgia , Laringectomia , Doenças Faríngeas/epidemiologia , Complicações Pós-Operatórias/epidemiologia , Hemoglobinas/metabolismo , Humanos , Neoplasias Laríngeas/patologia , Neoplasias Laríngeas/radioterapia , Laringe/patologia , Fatores de Risco
15.
J Biosci Bioeng ; 120(2): 210-7, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25622768

RESUMO

Mouse embryonic fibroblasts (MEFs) are commonly used as feeder cells for the generation of human induced pluripotent stem cells (hiPSCs). However, medical applications of cell derivatives of hiPSCs generated with a MEF feeder system run the risk of having xeno-factor contamination due to long-term cell culturing under an animal factor-containing environment. We developed a new method for the derivation of human fibroblast-like cells (FLCs) from a previously established hiPSC line in an FLC differentiation medium. The method was based on direct differentiation of hiPSCs seeded on Matrigel followed by expansion of differentiating cells on gelatin. Using inactivated FLCs as feeder layers, primary human foreskin fibroblasts were successfully reprogrammed into a state of pluripotency by Oct4, Sox2 Klf4, and c-Myc (OSKM) transcription factor genes, with a reprogramming efficiency under an optimized condition superior to that obtained on MEF feeder layers. Furthermore, the FLCs were more effective in supporting the growth of human pluripotent stem cells. The pluripotency and differentiation capability of the cells cultured on FLC feeder layers were well retained. Our results suggest that FLCs are a safe alternative to MEFs for hiPSC generation and expansion, especially in the clinical settings wherein hiPSC derivatives will be used for medical treatment.


Assuntos
Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células Alimentadoras/citologia , Fibroblastos/citologia , Células-Tronco Pluripotentes Induzidas/citologia , Animais , Proliferação de Células , Separação Celular , Células Cultivadas , Reprogramação Celular , Humanos , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/genética , Camundongos , Fator 3 de Transcrição de Octâmero/genética , Células-Tronco Pluripotentes/citologia , Proteínas Proto-Oncogênicas c-myc/genética , Fatores de Transcrição SOXB1/genética
16.
Dig Dis Sci ; 59(4): 778-86, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24162270

RESUMO

BACKGROUND: The early diagnosis of pancreas allograft dysfunction is crucial for the management and long-term survival of transplanted pancreases. We investigated whether intercellular adhesion molecular-1 (ICAM-1), Fas, and Fas ligand (FasL) can be used as novel biomarkers of acute pancreaticoduodenal allograft dysfunction in pigs. METHODS: Forty outbred landraces were randomly divided into three groups. In the control group (8 pigs), a sham operation was performed but no drugs were administered. In groups 1 and 2 (8 pairs each), pancreaticoduodenal transplantation was performed, with the latter administered immunosuppressive drugs and the former not administered drugs. The expression of ICAM-1, Fas, and FasL mRNA in the peripheral vein blood was assessed by flow cytometry and RT-PCR, pre-transplant and on days 1, 3, 5, and 7 after transplantation. Simultaneously, the levels of glucose, insulin, and glucagon in the serum of the recipients were evaluated. The allograft pancreas tissue was obtained to assess the pathological damage and the expression of Fas and FasL by immunohistochemistry. RESULTS: On the first 7 days after transplantation, ICAM-1, Fas, and FasL mRNA expression in the blood leukocytes of the recipient increased significantly in groups 1 and 2 compared with the control group (P < 0.01). However, the levels in group 2 were significantly lower than those in group 1 (P < 0.05). Interestingly, the FasL expression increased but the Fas expression decreased gradually in the graft pancreas tissue during the first week after transplantation in both groups 1 and 2 compared with the control group (P < 0.05). The levels of serous glucose, insulin, and glucagon in groups 1 and 2 obviously changed on day 1 after transplantation but returned to normal on day 2. The recipient's pancreas pathological sections did not exhibit any rejection changes on days 1 and 3 after transplantation but showed rejection damage on days 5 and 7. CONCLUSION: ICAM-1, Fas, and FasL were found to be sensitive biomarkers of acute pancreas allograft dysfunction after pancreaticoduodenal transplantation in pigs, and their monitoring could be used to evaluate the effectiveness of the immunosuppression therapy.


Assuntos
Biomarcadores/sangue , Proteína Ligante Fas/sangue , Rejeição de Enxerto/diagnóstico , Molécula 1 de Adesão Intercelular/sangue , Receptor fas/sangue , Aloenxertos , Animais , Duodeno/transplante , Glucagon/sangue , Rejeição de Enxerto/patologia , Insulina/sangue , Leucócitos/química , Pâncreas/patologia , Transplante de Pâncreas , Suínos
17.
Acta Biomater ; 9(8): 7679-90, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23669619

RESUMO

A series of amphiphilic 4- and 6-armed star triblock co-polymers poly(ε-caprolactone)-b-poly(2-(diethylamino)ethyl methacrylate)-b-poly(poly(ethylene glycol) methyl ether methacrylate) (4/6AS-PCL-b-PDEAEMA-b-PPEGMA) were developed by a combination of ring opening polymerization and continuous activators regenerated by electron transfer atom transfer radical polymerization. The critical micelle concentration values of the star co-polymers in aqueous solution were extremely low (2.2-4.0mgl(-1)), depending on the architecture of the co-polymers. The self-assembled blank and doxorubicin (DOX)-loaded three layer micelles were spherical in shape with an average size of 60-220nm determined by scanning electron microscopy and dynamic light scattering. The in vitro release behavior of DOX from the three layer micelles exhibited pH-dependent properties. The DOX release rate was significantly accelerated by decreasing the pH from 7.4 to 5.0, due to swelling of the micelles at lower pH values caused by the protonation of tertiary amine groups in DEAEMA in the middle layer of the micelles. The in vitro cytotoxicity of DOX-loaded micelles to HepG2 cells suggested that the 4/6AS-PCL-b-PDEAEMA-b-PPEGMA micelles could provide equivalent or even enhanced anticancer activity and bioavailability of DOX and thus a lower dosage is sufficient for the same therapeutic efficacy. The results demonstrate that the pH-sensitive multilayer micelles could have great potential application in delivering hydrophobic anticancer drugs for improved cancer therapy.


Assuntos
Sobrevivência Celular/efeitos dos fármacos , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/química , Doxorrubicina/administração & dosagem , Metacrilatos/química , Nanocápsulas/administração & dosagem , Nylons/química , Polietilenoglicóis/química , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Cristalização/métodos , Difusão , Doxorrubicina/química , Células Hep G2 , Humanos , Concentração de Íons de Hidrogênio , Teste de Materiais , Micelas , Nanocápsulas/química , Nanocápsulas/ultraestrutura , Tamanho da Partícula , Poliésteres , Ácidos Polimetacrílicos
18.
Inflammation ; 36(5): 1087-93, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23588345

RESUMO

The aim of this study was to explore the association between the single-nucleotide polymorphisms of interleukin-1 receptor-associated kinase-M (IRAK-M) gene and the susceptibility of sepsis. The allele frequency and genotype distribution of IRAK-M gene polymorphisms were assessed in 118 controls and 82 sepsis patients by semiquantitative polymerase chain reaction and restriction fragment length polymorphism (RFLP) analysis. The plasma levels of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were detected by enzyme-linked immunosorbent assay. Associations between IRAK-M polymorphisms and the susceptibility of sepsis were analyzed by Cox regression. Data were analyzed by the χ(2) test and the Student's t test, whenever appropriate. Statistical calculations were performed by using statistical package SPSS version 18.0. The genotype distribution of IRAK-M+22148 polymorphism significantly differed between the sepsis and control groups (P < 0.0001). The frequency of the G allele was remarkably more common in the sepsis group than that of the control group (P < 0.0001). However, the frequency of the A allele was significantly less common in the sepsis group than that of control group (P < 0.0001). Moreover, the plasma levels (in picograms per milliliter) of TNF-α and IL-6 in patients with G/G genotype were greatly higher than those with A/A genotype after lipopolysaccharide stimulation (P < 0.05). The genetic polymorphism of IRAK-M+22148 G>A is associated with the susceptibility of sepsis. The G/G genotype of IRAK-M increases the risk of developing sepsis, and the A/A genotype may play a protective role in the process of developing sepsis.


Assuntos
Quinases Associadas a Receptores de Interleucina-1/genética , Interleucina-6/sangue , Sepse/genética , Fator de Necrose Tumoral alfa/sangue , Adulto , Idoso , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Mediadores da Inflamação/sangue , Lipopolissacarídeos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Sepse/microbiologia
19.
PLoS One ; 8(12): e83964, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24386317

RESUMO

This prospective study compares different clinical characteristics and outcomes of patients with two types of sacral extradural spinal meningeal cysts (SESMC) undergoing different means of surgical excision. Using the relationship between the cysts and spinal nerve roots fibers (SNRF) as seen under microscope, SESMCs were divided into two types: cysts with SNRF known as Tarlov cysts and cysts without. The surgical methods were tailored to the different types of SESMCs. The improved Japanese Orthopedic Association (IJOA) scoring system was used to evaluate preoperative and postoperative neurological function of the patients. Preoperative IJOA scores were 18.5 ± 1.73, and postoperative IJOA scores were 19.6 ± 0.78. The difference between preoperative and postoperative IJOA scores was statistically significant (t = -4.52, p = 0.0001), with a significant improvement in neurological function after surgery. Among the improvements in neurological functions, the most significant was sensation (z=-2.74, p=0.006), followed by bowel/bladder function (z=-2.50, p=0.01). There was a statistically significant association between the types of SESMC and the number (F=12.57, p=0.001) and maximum diameter (F=8.08, p=0.006) of the cysts. SESMC with SNRF are often multiple and small, while cysts without SNRF tend to be solitary and large. We advocate early surgical intervention for symptomatic SESMCs in view of significant clinical improvement postoperatively.


Assuntos
Sacro , Cistos de Tarlov/cirurgia , Adolescente , Adulto , Idoso , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Recuperação de Função Fisiológica , Nervos Espinhais/fisiopatologia , Cistos de Tarlov/diagnóstico , Cistos de Tarlov/fisiopatologia , Fatores de Tempo , Resultado do Tratamento , Cicatrização , Adulto Jovem
20.
Beijing Da Xue Xue Bao Yi Xue Ban ; 44(4): 599-601, 2012 Aug 18.
Artigo em Chinês | MEDLINE | ID: mdl-22898855

RESUMO

OBJECTIVE: To prospectively analyze the clinical features and characteristics of multi-segments intramedullary spinal cord tumors in adolescent patients. METHODS: In our study, 25 consecutive adolescent patients with multi-segments intramedullary spinal cord tumors were recruited, who underwent microsurgery for the tumor using a posterior approach and were hospitalized in Peking University Third Hospital within a period of 8 years. The tumor was exposed through dorsal myelotomy. Preoperative and postoperative neurological functions were scored using the improved Japanese orthopaedic association score system (IJOA) grading system. The functional outcome was defined as postoperative IJOA score minus preoperative IJOA score. All the patients were followed-up until Oct. 30, 2011. RESULTS: There were 15 male and 10 female adolescent patients younger than 25 years. Their mean age was (15.3±6.83) years. The most common initial symptom was sensory disturbance (including pain and/or numbness, 52%, 13/25), followed by motor disturbance (including limbs weakness and gait deterioration, 24%, 6/25), pain and motor disturbance (12%, 3/25), as well as fever, limbs deformities, and sphincter dysfunction, respectively. The preoperative IJOA scores of the patients were (14.4±3.38). The postoperative IJOA scores of the patients were (15.5±3.31). The most commonly involved location was the cervicothoracic segments (36%, 9/25), followed by the conus terminalis (24%, 6/25), the cervical region(16%, 4/25), the thoracic region (16%, 4/25), and the lumbus region (8%, 2/25). The average involved segments were (4.4±1.38). The most frequent tumors were neurodevelopmental tumors (including lipoma, epidermoid cyst and teratoma) (32%, 8/25), followed by astrocytomas (28%, 7/25), ependymomas (20%, 5/25), hemangioblastomas (12%, 3/25), and glioblastomas and schwannomas, respectively. CONCLUSION: In adolescent patients with multi-segments intramedullary spinal cord tumors, the most commonly involved locations are the cervicothoracic segments and the conus terminalis, while the most frequent tumors are neurodevelopmental tumors and astrocytomas. Good prognosis in adolescent patients is observed in a long-term follow-up.


Assuntos
Astrocitoma/cirurgia , Lipoma/cirurgia , Neoplasias da Medula Espinal/cirurgia , Adolescente , Vértebras Cervicais , Ependimoma/cirurgia , Feminino , Humanos , Vértebras Lombares , Masculino , Neoplasias da Medula Espinal/patologia , Vértebras Torácicas , Adulto Jovem
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