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OBJECTIVE: To investigate the role of ginsenoside Rd (GRd) in acute myeloid leukemia (AML) cell differentiation. METHODS: AML cells were treated with GRd (25, 50, 100 and 200 µg/mL), retinoic acid (RA, 0.1g/L) and PD98059 (20 mg/mL) for 72 h, cell survival was detected by methylthiazolyldiphenyl-tetrazolium bromide and colony formation assays, and cell cycle was detected by flow cytometry. Cell morphology and differentiation were observed by Wright-Giemsa staining, peroxidase chemical staining and cellular immunochemistry assay, respectively. The protein expression levels of GATA binding protein 1 (GATA-1), purine rich Box-1 (PU.1), phosphorylated-extracellular signal-related kinase (p-ERK), ERK, phosphorylated-glycogen synthase kinase-3ß (p-GSK3ß), GSK3ß and signal transducer and activator of transcription 1 (STAT1) were detected by Western blot. Thirty-six mice were randomly divided into 3 groups using a random number table: model control group (non-treated), GRd group [treated with 200 mg/(kg·d) GRd] and homoharringtonine (HTT) group [treated with 1 mg/(kg·d) HTT]. A tumor-bearing nude mouse model was established, and tumor weight and volume were recorded. Changes of subcutaneous tumor tissue were observed after hematoxylin and eosin staining. WT1 and GATA-1 expressions were detected by immunohistochemical staining. RESULTS: The cell survival was inhibited by GRd in a dose-dependent manner and GRd caused G0/G1 cell arrest (p<0.05). GRd treatment induced leukemia cell differentiation, showing increased expressions of peroxidase and specific proteins concerning erythrogenic or granulocytic differentiation (p<0.05). GRd treatment elicited upregulation of p-ERK, p-GSK-3ß and STAT1 expressions in cells, and reversed the effects of PD98059 on inhibiting the expressions of peroxidase, GATA-1 and PU.1 (P<0.05). After GRd treatment, tumor weight and volume of mice were decreased, and tumor cells underwent massive apoptosis and necrosis (P<0.05). WT1 level was decreased, and GATA-1 level was significantly increased in subcutaneous tumor tissues (P<0.05 or P<0.01). CONCLUSION: GRd might induce the differentiation of AML cells via regulating the ERK/GSK-3ß signaling pathway.
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An efficient Ni-catalyzed hydrodifluoroalkylation of unactivated alkenes with bromodifluoroacetate by using PhSiH3 as hydride source was developed. The transformation affords aliphatic difluorides with anti-Markovnikov regioselectivity. A wide range of highly remote alkenes, simple alkenes, drug molecules, commercially available CF2 precursors, and even nonfluorinated substrates are competent in this reaction under mild conditions, demonstrating the practicability of the strategy. Moreover, mechanistic studies indicated that the difluoroalkyl radical might be a key intermediate to this transformation.
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OBJECTIVE: To investigate the role of petroleum ether extract of Rhizoma Amorphophalli (SLG) in inhibiting proliferation and promoting apoptosis and differentiation of leukemia K562 cells. METHODS: K562 cells were processed by SLG and PD98059 which was the ERK signaling pathway blocker. Then cell vitality was tested by MTT. Cell apoptosis rate and positive percentage of antigen expression related with differentiation were detected by flow cytometry. The protein expression levels of ERK1/2 and pîERK1/2 were detected by Western blot. RESULTS: The proliferation activity of K562 was reduced by 50, 100, 200 mg/L SLG in a concentration dependent manner (r=0.9997). The apoptosis rate and positive expression rate of CD11b, CD14 and CD42b which were related with differentiation were raised by SLG, as well as the expression of pîERK1/2, while PD98059 could reverse the promoting effect of SLG on apoptosis and differentiation partially. CONCLUSION: SLG can inhibit the proliferation and promote apoptosis and differentiation of K562 cells through ERK signaling pathway.
Assuntos
Petróleo , Alcanos , Apoptose , Proliferação de Células , Humanos , Células K562 , Extratos Vegetais/farmacologiaRESUMO
OBJECTIVE: To investigate the potential efficacy of panaxadiol saponins component (PDS-C) in the treatment of aplastic anemia (AA) model mice. METHODS: Totally 70 mice were divided into 7 groups as follows: normal, model, low-, medium-, high-dose PDS-C (20, 40, 80 mg/kg, namely L-, M-, H-PDS-C), cyclosporine (40 mg/kg), and andriol (25 mg/kg) groups, respectively. An immune-mediated AA mouse model was established in BALB/c mice by exposing to 5.0 Gy total body irradiation at 1.0 Gy/min, and injecting with lymphocytes from DBA mice. On day 4 after establishment of AA model, all drugs were intragastrically administered daily for 15 days, respectively, while the mice in the normal and model groups were administered with saline solution. After treatment, the peripheral blood counts, bone marrow pathological examination, colony forming assay of bone marrow culture, T lymphocyte subpopulation analysis, as well as T-bet, GATA-3 and FoxP3 proteins were detected by flow cytometry and Western blot. RESULTS: The peripheral blood of white blood cell (WBC), platelet, neutrophil counts and hemoglobin (Hb) concentration were significantly decreased in the model group compared with the normal group (all P<0.01). In response to 3 dose PDS-C treatment, the WBC, platelet, neutrophil counts were significantly increased at a dose-dependent manner compared with the model group (all P<0.01). The myelosuppression status of AA was significantly reduced in M-, H-PDS-C groups, and hematopoietic cell quantity of bone marrow was more abundant than the model group. The colony numbers of myeloid, erythroid and megakaryocytic progenitor cells in the model group were less than those of the normal mice in bone marrow culture, while, PDS-C therapy enhanced proliferation of hematopoietic progenitor cells by significantly increasing colony numbers (all P<0.01). Furthermore, PDS-C therapy increased peripheral blood CD3+ and CD3+CD4+ cells and reduced CD3+CD8+ cells (P<0.05 or P<0.01). Meanwhile, PDS-C treatment at medium- and high doses groups also increased CD4+CD25+FoxP3+ cells, downregulated T-bet protein expression, and upregulated GATA-3 and FoxP3 protein expressions in spleen cells (P<0.05). CONCLUSION: PDS-C possesses dual activities, promoting proliferation hematopoietic progenitor cells and modulating T lymphocyte immune functions in the treatment of AA model mice.
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Anemia Aplástica/tratamento farmacológico , Ginsenosídeos/farmacologia , Hematopoese/efeitos dos fármacos , Panax , Saponinas/farmacologia , Linfócitos T/efeitos dos fármacos , Anemia Aplástica/sangue , Animais , Modelos Animais de Doenças , Camundongos , Camundongos Endogâmicos BALB CRESUMO
MicroRNA (miR)155 has a crucial role in various cellular functions, including differentiation of hematopoietic cells, immunization, inflammation and cardiovascular diseases. The present study aimed to investigate the roles and mechanisms of miR155 in treatmentresistant depression (TRD). A Cell Counting Kit8 assay and flow cytometry were performed to assess the cell viability and apoptosis of microglial cells, respectively. Western blotting and reverse transcriptionquantitative polymerase chain reaction assays were used to evaluate the associated protein and mRNA expression, respectively. The results revealed that miR155 reduced the cell viability of BV2 microglial cells, and miR155 enhanced the expression levels of proinflammatory cytokines in BV2 microglial cells. Furthermore, conditioned medium from miR155treated microglia decreased the cell viability of HT22 hippocampal cells. miR155treated microglia increased the apoptosis of neuronal hippocampal cells by modulating the expression levels of apoptosis regulator Bax, apoptosis regulator Bcl2, procaspase3 and cleavedcaspase3. The cell cycle distribution was disrupted by miR155treated microglia through induction of S phase arrest. Furthermore, the overexpression of suppressor of cytokine signaling 1 reversed the proapoptotic effect of activated microglia on hippocampal neuronal cells. In conclusion, the present results suggested that miR155 mediated the inflammatory injury in hippocampal neuronal cells by activating the microglial cells. The potential effects of miR155 on the activation of microglial cells suggest that miR155 may be an effective target for TRD therapies.
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Inflamação/etiologia , Inflamação/metabolismo , MicroRNAs/genética , Microglia/imunologia , Microglia/metabolismo , Células Piramidais/metabolismo , Animais , Apoptose/genética , Ciclo Celular/genética , Sobrevivência Celular/genética , Células Cultivadas , Citocinas/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Camundongos , Transdução de Sinais , Proteína 1 Supressora da Sinalização de Citocina/metabolismoRESUMO
OBJECTIVE: To investigate the potential efficacy of panaxadiol saponins component (PDS-C), a biologically active fraction isolated from total ginsenosides, to reverse chemotherapy-induced myelosuppression and pancytopenia caused by cyclophamide (CTX). METHODS: Mice with myelosuppression induced by CTX were treated with PDS-C at a low- (20 mg/kg), moderate- (40 mg/kg), or high-dose (80 mg/kg) for 7 consecutive days. The level of peripheral white blood cell (WBC), neutrophil (NEU) and platelet (PLT) were measured, the histopathology and colony formation were observed, the protein kinase and transcription factors in hematopoietic cells were determined by immunohistochemical staining and Western blot. RESULTS: In response to PDS-C therapy, the peripheral WBC, NEU and PLT counts of CTX-induced myelosuppressed mice were significantly increased in a dose-dependent manner. Similarly, bone marrow histopathology examination showed reversal of CTX-induced myelosuppression with increase in overall bone marrow cellularity and the number of hematopoietic cells (P<0.01). PDS-C also promoted proliferation of granulocytic and megakaryocyte progenitor cells in CTX-treated mice, as evidenced by significantly increase in colony formation units-granulocytes/monocytes and -megakaryocytes (P<0.01). The enhancement of hematopoiesis by PDS-C appears to be mediated by an intracellular signaling pathway, this was evidenced by the up-regulation of phosphorylated mitogen-activated protein kinase (p-MEK) and extracellular signal-regulated kinases (p-ERK), and receptor tyrosine kinase (C-kit) and globin transcription factor 1 (GATA-1) in hematopoietic cells of CTX-treated mice (P<0.05). CONCLUSIONS: PDS-C possesses hematopoietic growth factor-like activities that promote proliferation and also possibly differentiation of hematopoietic progenitor cells in myelosuppressed mice, probably mediated by a mechanism involving MEK and ERK protein kinases, and C-kit and GATA-1 transcription factors. PDS-C may potentially be a novel treatment of myelosuppression and pancytopenia caused by chemotherapy.
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Antineoplásicos/efeitos adversos , Ciclofosfamida/efeitos adversos , Ginsenosídeos/uso terapêutico , Hematopoese/efeitos dos fármacos , Células Mieloides/patologia , Panax/química , Pancitopenia/tratamento farmacológico , Saponinas/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fator de Transcrição GATA1/metabolismo , Ginsenosídeos/farmacologia , Camundongos , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Células Mieloides/efeitos dos fármacos , Pancitopenia/induzido quimicamente , Pancitopenia/patologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-kit/metabolismo , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND: Side-population (SP) cells, identified by their capacity to efflux Hoechst dye, are highly enriched for stem/progenitor cell activity. They are found in many mammalian tissues, including mouse heart. Studies suggest that cardiac SP (CSP) cells can be divided into SCA1+/CD31-, SCA1+/CD31+ and SCA1-/CD31- CSP subpopulations. SCA1+/CD31- were shown to be cardiac and endothelial stem/progenitors while SCA1+/CD31+ CSP cells are endothelial progenitors. SCA1-/CD31- CSP cells remain to be fully characterized. In this study, we characterized SCA1-/CD31- CSP cells in the adult mouse heart, and investigated their abilities to proliferate, differentiate and migrate in vitro and in vivo. METHODS AND RESULTS: Using fluorescence-activated cell sorting, reverse transcriptase/polymerase chain reaction, assays of cell proliferation, differentiation and migration, and a murine model of myocardial infarction we show that SCA1-/CD31- CSP cells are located in the heart mesenchyme and express genes characteristic of stem cells and endothelial progenitors. These cells were capable of proliferation, differentiation, migration and vascularization in vitro and in vivo. Following experimental myocardial infarction, the SCA1-/CD31- CSP cells migrated from non-infarcted areas to the infarcted region within the myocardium where they differentiated into endothelial cells forming vascular (tube-like) structures. We further demonstrated that the SDF-1α/CXCR4 pathway may play an important role in migration of these cells after myocardial infarction. CONCLUSIONS: Based on their gene expression profile, localization and ability to proliferate, differentiate, migrate and vascularize in vitro and in vivo, we conclude that SCA1-/CD31- CSP cells may serve as endothelial progenitor cells in the adult mouse heart.
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Ataxina-1/fisiologia , Células Endoteliais/fisiologia , Infarto do Miocárdio/patologia , Miócitos Cardíacos/fisiologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/fisiologia , Células da Side Population/fisiologia , Animais , Técnicas de Cultura de Células , Diferenciação Celular , Movimento Celular , Proliferação de Células , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/etiologiaRESUMO
OBJECTIVE: To investigate the role and significance of T help cells 17(Th17) in pathogenesis of idiopathic thrombocytopenic purpura (ITP). METHODS: Peripheral blood samples from ITP patients and normal controls were examined for Th17 cell proportion by flow cytometry (FCM). Expression of IL-17, IL-23, IL-6 and TGF-ß1 in hematoplasma was detected by ELISA. The mRNA expression level of IL-17 and RORγt in peripheral blood mononuclear cells (PBMNC) from patients with ITP and normal controls were measured by RT-PCR technique, and expression levels of pSTAT3 and RORγt proteins were analyzed by Western-blot. RESULTS: Th17 cells in peripheral blood from patients with ITP was greatly increased when compared with normal control group (P<0.05). Expressions of IL-17, IL-23, IL-6 and TGF-ß1 in hematoplasma of ITP patients were all significantly higher than those in normal control group (all P<0.01). mRNA expression levels of IL-17 and RORγt in PBMNC from patients with ITP were much higher than those in normal controls (P<0.05). Protein expressions of pSTAT3 and RORγt in PBMNC of ITP patients were greatly increased as compared with those in control (P<0.05). CONCLUSION: Th17 cell subgroup may play a role in incidence and development of ITP, which may participate in the pathogenesis of ITP by increasing Th17 cell proportion and altering the expression level of Th17-related cytokines as well as regulatory and transcriptional factors.
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Leucócitos Mononucleares , Púrpura Trombocitopênica Idiopática , Contagem de Células , Citocinas , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares , RNA Mensageiro , Células Th17 , Fator de Crescimento Transformador beta1RESUMO
OBJECTIVE: To investigate the effects of panaxadiol saponins component (PDS-C) isolated from total saponins of panax ginseng on proliferation, differentiation and corresponding gene expression profile of megakaryocytes. METHODS: Bone marrow culture of colony forming assay of megakaryocytic progenitor cells (CFU-MK) was observed for the promoting proliferation mediated by PDS-C, and differentiation of megakaryocytic blasts caused by PDS-C was analyzed with flow cytometry in CHRF-288 and Meg-01 cells, as well as proliferation, differentiation-related genes expression profile and protein expression levels were detected by human gene expression microarray and western blot. RESULTS: In response to PDS-C 10, 20 and 50 mg/L, CFU-MK from 10 human bone marrow samples was increased by 28.9%±2.7%, 41.0%±3.2% and 40.5%±2.6% over untreated control, respectively (P <0.01, each). Flow cytometry analysis showed that PDS-C treated CHRF-288 cells and Meg-01 cells significantly increased in CD42b, CD41, TSP and CD36 positive ratio, respectively. PDS-C induced 29 genes up-regulated more than two-fold commonly in both cells detected by human expression microarray representing 4000 known genes. The protein expression levels of ZNF91, c-Fos, BTF3a, GATA-1, RGS2, NDRG2 and RUNX1 were increased with western blot in correspond to microarray results. CONCLUSION: PDS-C as an effective component for hematopoiesis, play the role to enhance proliferation and differentiation of megakaryocytes, also up-regulated expression of proliferation, differentiation-related genes and proteins in vitro.
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Diferenciação Celular/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Perfilação da Expressão Gênica , Ginsenosídeos/farmacologia , Megacariócitos/citologia , Megacariócitos/metabolismo , Patentes como Assunto , Saponinas/farmacologia , Western Blotting , Células da Medula Óssea/citologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Citometria de Fluxo , Humanos , Megacariócitos/efeitos dos fármacos , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Fatores de Transcrição/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
OBJECTIVE: Bone metastasis was common in patients with malignant tumors. The purpose of this study was to investigate the serum bone-specific alkaline phosphatase (B-ALP) as a biomarker in the diagnosis of osseous metastases in patients with cancers. METHODS: We searched the databases of Pubmed, Cochrane Library, Medline, CNKI and Wanfang to screen the relevant articles about the serum B-ALP detection in the diagnosis of osseous metastases in patients with malignant carcinomas. The pooled sensitivity, specificity, summary receiver operating characteristic (SROC) curve were calculated by STATA12.0 software. RESULTS: Nineteen trials with 3 268 subjects were finally included in this study. The mean level of serum B-ALP was 41.50 ± 26.61 µg/L (216.90 ± 139.00U/L) in patients with osseous metastases and 14.49 ± 5.52 µg/L (103.30 ± 39.44 U/L) in patients without osseous metastases. The serum level of B-ALP was significant higher in the osseous metastases group than that in the control group (P < 0.05); The pooled sensitivity and specificity for diagnosis of osseous metastases were 0.74 with its 95% confidence interval (95% CI) of 0.62-0.83 and 0.80 (95% CI: 0.67-0.89), respectively. The area under the SRCO was 0.86 (95% CI: 0.83-0.89). CONCLUSION: Serum B-ALP can be a promising biomarker for detection of osseous metastases in patients with cancers.
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Fosfatase Alcalina/sangue , Biomarcadores Tumorais/sangue , Neoplasias Ósseas/sangue , Neoplasias Ósseas/diagnóstico , Carcinoma/sangue , Carcinoma/patologia , Metástase Neoplásica/diagnóstico , Neoplasias Ósseas/patologia , Osso e Ossos/patologia , Estudos de Casos e Controles , Humanos , Metástase Neoplásica/patologia , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To study the clinical significance of tidal breathing lung function test in 1-4 years old children with wheezing diseases. METHODS: A total of 141 1-4 years old children with wheezing diseases were enrolled as the observed groups (41 cases of asthma, 54 cases of asthmatic bronchitis, and 46 cases of bronchopneumonia). Thirty children without respiratory diseases were enrolled as the control group. All the recruits underwent tidal breathing lung function test. The observed groups underwent bronchial dilation test, and tidal breathing flow volume (TBFV) parameters were evaluated before and after bronchial dilation test. RESULTS: The observed groups showed obstructive ventilatory disorder (65%) according to the TBFV loop, and their ratio of time to peak tidal expiratory flow (TPTEF) to total expiratory time (TE) and ratio of volume to peak expiratory flow (VPEF) to total expiratory volume (VE) were significantly lower than in the control group (P<0.05). The asthma subgroup had significantly improved TPTEF/TE and VPEF/VE after bronchial dilation test (P<0.05). Taking an improvement rate of ≥ 15% either for TPTEF/TE or for VPEF/VE as an indicator of positive bronchial dilation test, the bronchial dilation test had a sensitivity of 47% and a specificity of 84% in diagnosing asthma in 1-4 years old children. The positive rate was 28% among the children in the asthma subgroup with an TPTEF/TE ratio of ≥ 23% before bronchial dilation test, versus 65% in those with an TPTEF/TE ratio of <23%. CONCLUSIONS: Obstructive ventilatory disorder is the main impairment of tidal breathing lung function in 1-4 years old children with wheezing diseases. Tidal breathing bronchial dilation test can reflect a reversal of airway obstruction to a certain extent. The sensitivity of bronchial dilation test for the diagnosis of asthma is not satisfactory in 1-4 years old children with wheezing diseases, but this test has a relatively high diagnostic value in children with severe airway obstruction.
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Bronquite/diagnóstico , Respiração , Testes de Função Respiratória/métodos , Sons Respiratórios/diagnóstico , Asma/diagnóstico , Asma/fisiopatologia , Bronquite/fisiopatologia , Broncopneumonia/diagnóstico , Broncopneumonia/fisiopatologia , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Sons Respiratórios/efeitos dos fármacos , Sons Respiratórios/fisiopatologiaRESUMO
OBJECTIVE: To investigate the effects of sodium copper chlorophyllin (SCC) on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells (MSCs) from mice with aplastic anemia. METHODS: A mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2×10(6) lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups: the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A (CsA); and an untreated model control group (model group); while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts (CFU-F) in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia. RESULTS: Peripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 (FOXP3(+)) T cells was increased in T cell-MSC cocultures, and the cytokine transforming growth factor ß1 was up-regulated in SCC-treated groups. CONCLUSION: The results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with aplastic anemia.
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Anemia Aplástica/terapia , Clorofilídeos/farmacologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Anemia Aplástica/sangue , Anemia Aplástica/patologia , Animais , Antraquinonas/metabolismo , Biomarcadores/metabolismo , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/patologia , Cálcio/metabolismo , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Feminino , Terapia de Imunossupressão , Contagem de Leucócitos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Osteoblastos/efeitos dos fármacos , Osteoblastos/patologia , Contagem de Plaquetas , Linfócitos T/efeitos dos fármacosRESUMO
OBJECTIVE: To investigate the effects of Panax notoginseng saponins (PNS) on the proliferation and differentiation in NIH3T3 cells. METHODS: NIH3T3 cells were treated by various concentrations of PNS 0, 0.05, 0.10, 0.20, and 0.40 g/L. The vitality and proliferation potential of cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, the alkaline phosphatase (ALP) activity was measured by p-nitrophenyl phosphate (pNPP) assay, and the mineralization formation ability was tested for the cellular differentiation toward osteoblast, as well as the expression level of phosphorylated extracellular signal-regulated kinase1/2(P-ERK1/2), extracellular signal-regulated kinase1/2 (ERK1/2) protein kinase was analyzed by Western blot with total cell lysate of NIH3T3 cells treated by PNS. RESULTS: Both MTT and pNPP assay showed that optical density (OD) values were increased in response to PNS treatment at a dose-dependent pattern. The mineralization formation ability was enhanced in PNS-treated NIH3T3 cells compared with untreated cells. Meanwhile, the expression level of P-ERK1/2 protein kinase was up-regulated in PNS-treated NIH3T3 cells, while, the expression level of ERK1/2 protein kinase revealed no obvious difference with or without PNS treated cells. CONCLUSION: PNS could pay a role to promote the proliferation and differentiation in NIH3T3 cells by means of up-regulation of P-ERK1/2 protein kinase.
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Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Panax notoginseng/química , Saponinas/farmacologia , Fosfatase Alcalina/metabolismo , Animais , Cálcio/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fibroblastos/enzimologia , Camundongos , Células NIH 3T3 , Osteocalcina/metabolismoRESUMO
OBJECTIVE: To study the prognosis of childhood asthma and the factors influencing asthmatic attacks and prognosis. METHODS: The medical data of 212 children with asthma who were followed up for more than 5 years were retrospectively studied. RESULTS: During the 5-year follow up, asthmatic attacks termination was found in 121 cases (57.1%) and asthma persistence was observed in 91 cases. Respiratory tract infections were found as the major factors inducing asthmatic attacks (71.7%), followed by inhaled allergens (17.0%).The children with asthma induced by respiratory tract infections had a higher remission rate of asthmatic attacks (61.2%) than those induced by allergens (41.7%) or exercises (26.3%). Three risk factors for asthma persistence were identified: concurrent allergic rhinitis and eczema, parental asthma and allergy-induced wheezing. CONCLUSIONS: The 5-year follow-up study demonstrated that asthmatic attacks stopped in the majority of children with asthma. Respiratory tract infections may be the major factors inducing acute asthma attacks. The children with asthma induced by respiratory infections may experience a better outcome. Atopic children or children with the genetic background of atopy are at high risks for the development of persistent asthma.
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Asma/etiologia , Asma/tratamento farmacológico , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Lactente , Masculino , Prognóstico , Infecções Respiratórias/complicações , Estudos Retrospectivos , Fatores de RiscoRESUMO
In this paper, we determine the mutant W203F structure of TFsß-glucanase, which contains aromatic residue Trp203 at the active site of the enzyme. Residue Trp203 is stacked with the glucose product of cellotriose. Further analysis reveals that two extra calcium ions and a Tris molecule bind to the mutant structure. A Tris molecule, bound to the catalytic residues of Glu56 and Glu60, was found at the position normally taken by substrate binding at the -1 subsite. In addition, a second Ca(2+) ion was found near the residues Phe152 and Glu154 on the protein's surface, and a third one near the active site residue Asp202. Kinetic experiments reveal that both Tris and imidazole are competitive inhibitors, while calcium is a noncompetitive inhibitor for TFsß-glucanase. The two types of enzymatic inhibition are first-time descriptions for the glycosyl hydrolase family 16.
Assuntos
Cálcio/farmacologia , Glicosídeo Hidrolases/antagonistas & inibidores , Glicosídeo Hidrolases/química , Trometamina/farmacologia , Cálcio/química , Domínio Catalítico/genética , Cátions Bivalentes/química , Cátions Bivalentes/farmacologia , Cristalografia por Raios X , Fibrobacter/enzimologia , Glicosídeo Hidrolases/genética , Imidazóis/química , Imidazóis/farmacologia , Mutação , Paenibacillus/enzimologia , Trometamina/química , Triptofano/química , Triptofano/genéticaRESUMO
This study was aimed to explore the inhibition effect and mechanism of hedyotis diffusa wild injection (HDI) on leukemia cell line (HL-60) in vitro. The leukemia cell line HL-60 was used as target cells. The inhibitory effects of HDI on proliferation of HL-60 cells were observed by MTT assay. The positive rate of cell apoptosis and the surface marker of granulocytic differentiation (CD33 and CD15) were measured by flow cytometry. The expressions of anti-apoptosis related gene (survivin and bcl-2) were detected by RT-PCR. The results showed that the growth of HL-60 cells was inhibited by higher concentration of HDI (3.12 - 12.5 ml/L) and inhibited obviously in dose-dependent manner (p < 0.05 or p < 0.01), but not suppressed by low concentration of HDI (1.56 ml/L) in liquid culture system (p > 0.05). The FCM and DNA Ladder results showed that the phenomenon of typical apoptosis did not detected after HL-60 cells were treated with the different concentrations of HDI for 24, 48 and 72 hours respectively. After HL-60 cells were treated with HDI (1.56, 3.12, 6.25 and 12.5 ml/L) for one week, the expression level of CD15 surface marker was all enhanced obviously. When treated with HDI (6.25 ml/L) for 3 weeks, the expression levels of survivin and bcl-2 gene were also decreased obviously by 60% and 44% respectively. It is concluded that HDI can inhibit HL-60 cells in the presence of its higher concentrations. The mechanisms of HDI may induce HL-60 cells differentiation, and suppress the expression of anti-apoptosis related gene (survivin or bcl-2) to inhibit the growth of HL-60 cells.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Células HL-60 , Humanos , Proteínas Inibidoras de Apoptose/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , SurvivinaRESUMO
Based on the continuous observation on located trees and branches, the phenology and sexual reproductive process of Xanthoceras sorbifolia Bunge in an annual growth cycle in Wudan area were investigated, and the relationships between fruit growth process of X. sorbifolia and effective accumulative temperature as well as the quantitative dynamics of flowers and fruits were studied. The results showed that the phenology of X. sorbifolia had a significant linear correlation with effective accumulative temperature, and there was a time sequence in the male flower silk elongating and antheral unfolding. The fruits of X. sorbifolia experienced three times of dropping during their growth process. The pollination rate was 26.4%, flower dropping rate was 73.6%, and fruit dropping rate was 92.9%, suggesting that the dropping of flowers and fruits was very serious. The fruits had a quicker vertical growth from early June to mid June and a quicker horizontal growth in the last ten-day of June, with the highest ratio of vertical to horizontal length being 2.17. The individuals of X. sorbifolia had definite differences in their phenology, which was disadvantageous to the cross pollination among the individuals, and the enhancement of pollination rate.
Assuntos
Ecossistema , Flores/crescimento & desenvolvimento , Frutas/crescimento & desenvolvimento , Sapindaceae/crescimento & desenvolvimento , Sapindaceae/fisiologia , China , Polinização , TemperaturaRESUMO
OBJECTIVE: To observe the effects of endothelial cells from umbilical cord blood (UCB) on the amplification of human early hematopoietic cells from UCB in vitro. METHODS: Endothelial cells from UCB were cultured by the optimized medium of endothelial cells. There were 2 experiment groups: cytokines group (SCF+IL-3+IL-6+GM-CSF, CKs group) and noncontact group (endothelial cell layer with CKs without contacting the CD34+ cells group). CD34+ cells from UCB were isolated by MiniMACS. After the cells in the CKs group and the noncontact group were cultured for 7 days, the amplifying folds of early hematopoietic cells were assayed. RESULTS: Early hematopoietic cells from UCB were expanded in the CKs group or the noncontact group. The amplifying folds of the noncontact group on early hematopoietic cells were significantly more than those of the CKs group. CONCLUSION: The amplification effect of the noncontact group on early hematopoietic cells is superior to that of the CKs group.