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1.
Cell Biol Int ; 2020 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-32339345

RESUMO

The dysregulation of proliferation and migration of vascular smooth muscle cells (VSMCs) contributes to atherosclerosis (AS) and accumulating reports indicate the crucial role of long noncoding RNA in AS. However, the role of small nucleolar RNA host gene 12 (SNHG12) in regulating the phenotypes of VSMCs and AS remains largely unknown. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to detect the expression levels of SNHG12 and miR-199a-5p in an in vivo AS model and VSMCs treated by oxidized low-density lipoprotein (ox-LDL). The proliferation ability, migration ability, and apoptosis of VSMCs were tested by cell counting kit-8, Transwell assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay, respectively. StarBase database was used to predict the binding sites between miR-199a-5p and SNHG12. The interaction between miR-199a-5p and SNHG12 was validated by qRT-PCR, western blot, and luciferase reporter assay. Western blot was used to examine the effects of SNHG12 and miR-199a-5p on the expression of hypoxia-inducible factor 1α (HIF-1α). We found that the expression level of SNHG12 was significantly increased in the animal model and VSMCs treated by ox-LDL. Knockdown of SNHG12 suppressed the proliferation and migration abilities of VSMCs, while overexpression of SNHG12 had the opposite effects. Mechanically, we validated that miR-199a-5p was a target of SNHG12, and the target gene of miR-199a-5p, HIF-1α could be indirectly and positively regulated by SNHG12. In conclusion, SHNG12 targeting miR-199a-5p/HIF-1α contributed to the pathophysiological process of AS by regulating the phenotypes of VSMCs, and could be a potential therapy target for this disease.

2.
J Glob Antimicrob Resist ; 20: 160-162, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31877398

RESUMO

OBJECTIVES: Recently, the Gram-negative bacterium Klebsiella oxytoca has been identified as an emerging pathogen. Here we report the draft genome of a 2,3-butanediol-producing strain, K. oxytoca CCTCC M207023, isolated from soil in Nanjing, China. The tetracycline-resistant phenotype and the high yield of 2,3-butanediol was demonstrated. METHODS: The draft genome of K. oxytoca CCTCC M207023 was determined using an Illumina NovaSeq™ 6000 next-generation DNA sequencing platform. Clean sequencing data were subsequently assembled using SOAPdenovo. RESULTS: The draft genome of K. oxytoca CCTCC M207023, comprising 5 658 144bp and with a GC content of 56.50%, was assembled into 5262 open-reading frames (ORFs). Antimicrobial resistance genes were also annotated. CONCLUSIONS: The draft genome sequence of K. oxytoca CCTCC M207023 reported here will be a reference for comparative analysis with the antimicrobial resistance mechanisms for the safety of 2,3-butanediol industrial production.

3.
Chin Med J (Engl) ; 132(21): 2601-2611, 2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31373906

RESUMO

BACKGROUND: In consideration of characteristics and functions, extra-cellular signal-regulated protein kinase 5 (ERK5) signaling pathway could be a new target for spinal cord injury (SCI) treatment. Our study aimed to evaluate the roles of ERK5 signaling pathway in secondary damage of SCI. METHODS: We randomly divided 70 healthy Wistar rats into five groups: ten in the blank group, 15 in the sham surgery + BIX02188 (sham + B) group, 15 in the sham surgery + dimethyl sulfoxide (DMSO; sham + D) group, 15 in the SCI + BIX02188 (SCI + B) group, and 15 in the SCI + DMSO (SCI + D) group. BIX02188 is a specific inhibitor of the ERK5 signaling pathway. SCI was induced by the application of vascular clips (with the force of 30 g) to the dura on T10 level, while rats in the sham surgery group underwent only T9-T11 laminectomy. BIX02188 or DMSO was intra-thecally injected at 1, 6, and 12 h after surgery or SCI. Spinal cord samples were taken for testing at 24 h after surgery or SCI. RESULTS: Expression of phosphorylated-ERK5 (p-ERK5) significantly increased after SCI. Application of BIX02188 indeed inhibited ERK5 signaling pathway and reduced the degree of spinal cord tissue injury, neutrophil infiltration and proinflammatory cytokine expression, nuclear factor-κB (NF-κB) activation and apoptosis (measured by TdT-mediated 2'-deoxyuridine 5'-triphosphate nick-end labeling, expression of Fas-ligand, BCL2-associated X [Bax], and B-cell lymphoma-2 [Bcl-2]). Double immunofluorescence revealed activation of ERK5 in neurons and microglia after SCI. CONCLUSION: ERK5 signaling pathway was activated in spinal neurons and microglia, contributing to secondary injury of SCI. Moreover, inhibition of ERK5 signaling pathway could alleviate the degree of SCI, which might be related to its regulation of infiltration of inflammatory cells and release of inflammatory cytokines, expression of NF-κB and cell apoptosis.

4.
Appl Microbiol Biotechnol ; 103(11): 4313-4324, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31016357

RESUMO

In recent years, eukaryotic microorganisms have been widely applied to offer many solutions for everyday life and have come to play important roles in agriculture, food, health care, and the fine-chemicals industry. However, the complex genetic background and low homologous recombination efficiency have hampered the implementation of large-scale and high-throughput gene editing in many eukaryotic microorganisms. The low efficiency of homologous recombination (HR) not only makes the modification process labor-intensive but also completely precludes the application of many otherwise very useful genome editing techniques. Thus, increasing the efficiency of HR is clearly an enabling technology for basic research and gene editing in eukaryotic microorganisms. In this review, we summarize the current strategies for enhancing the efficiency of HR in eukaryotic microorganisms (particularly yeasts and filamentous fungi), list some small molecules and candidate genes associated with homologous and non-homologous recombination, and briefly discuss the further development prospects of these strategies.


Assuntos
Fungos/genética , Edição de Genes/métodos , Recombinação Homóloga , Engenharia Metabólica/métodos , Leveduras/genética
5.
Bioresour Technol ; 281: 449-456, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30846235

RESUMO

Terpenoids are a large class of natural compounds based on the C5 isoprene unit, with many biological effects such activity against cancer and allergies, while some also have an agreeable aroma. Consequently, they have received extensive attention in the food, pharmaceutical and cosmetic fields. With the identification and analysis of the underlying natural product synthesis pathways, current microbial-based metabolic engineering approaches have yielded new strategies for the production of highly valuable terpenoids. Yarrowia lipolytica is a non-conventional oleaginous yeast that is rapidly emerging as a valuable host for the production of terpenoids due to its own endogenous mevalonate pathway and high oil production capacity. This review aims to summarize the status and strategies of metabolic engineering for the heterologous synthesis of terpenoids in Y. lipolytica in recent years and proposes new methods aiming towards further improvement of terpenoid production.


Assuntos
Engenharia Metabólica/métodos , Terpenos/metabolismo , Yarrowia/metabolismo , Produtos Biológicos/metabolismo
7.
ACS Synth Biol ; 8(2): 445-454, 2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30616338

RESUMO

The filamentous fungus Fusarium fujikuroi is well-known for its production of natural plant growth hormones: a series of gibberellic acids (GAs). Some GAs, including GA1, GA3, GA4, and GA7, are biologically active and have been widely applied in agriculture. However, the low efficiency of traditional genetic tools limits the further research toward making this fungus more efficient and able to produce tailor-made GAs. Here, we established an efficient CRISPR/Cas9-based genome editing tool for F. fujikuroi. First, we compared three different nuclear localization signals (NLS) and selected an efficient NLS from histone H2B (HTBNLS) to enable the import of the Cas9 protein into the fungal nucleus. Then, different sgRNA expression strategies, both in vitro and different promoter-based in vivo strategies, were explored. The promoters of the U6 small nuclear RNA and 5S rRNA, which were identified in F. fujikuroi, had the highest editing efficiency. The 5S rRNA-promoter-driven genome editing efficiency reached up to 79.2%. What's more, multigene editing was also explored and showed good results. Finally, we used the developed genome editing tool to engineer the metabolic pathways responsible for the accumulation of a series GAs in the filamentous fungus F. fujikuroi, and successfully changed its GA product profile, from GA3 to tailor-made GA4 and GA7 mixtures. Since these mixtures are more efficient for agricultural use, especially for fruit growth, the developed strains will greatly improve industrial GA production.


Assuntos
Sistemas CRISPR-Cas/genética , Fungos/genética , Fungos/metabolismo , Fusarium/genética , Fusarium/metabolismo , Giberelinas/metabolismo , Edição de Genes/métodos , Genoma Fúngico/genética
8.
Medicine (Baltimore) ; 97(27): e11140, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29979380

RESUMO

This study aims to explore the influence of bone resorption of the spinous process after single-segment interspinous process device (IPD) implantation on the biomechanics of the lumbar spine.The 3D finite element model of the lumbar spine (L3-L5) was modified, and 2 models that simulated the presence and absence of bone resorption of the spinous process were developed using an IPD (Wallis). Its biomechanical effects, such as change in range of motion (ROM) and intervertebral disc and facet stress, were introduced at operative (L4/5) and adjacent (L3/4) levels.Compared with the INT model, the Wallis model and Wallis-BR model had similar ROMs in lateral flexion and rotation. However, the Wallis model had a lower L3-5 ROM in flexion (20.4% lower) and extension (26.4% lower), and L4-L5 ROM in flexion (74.1% lower) and extension (70.8% lower), while the overall ROM of the Wallis-BR model was greater than that of the Wallis model. The stress on the L3/L4 intervertebral disc and facets was similar for all 3 models. Compared with the INT model and Wallis-BR model, the stress on the L4/L5 intervertebral disc and facets under all movements significantly decreased in the Wallis model. The stress on the L5 process was greater than that on the L4 process in both the Wallis model and Wallis-BR model, and the load on the processes that underwent bone resorption was lower than that of the Wallis model.The function of the IPD slowly decreased with the occurrence of bone resorption of the interspinous process. This bone remodeling may be associated with high stress after IPD implantation.


Assuntos
Fenômenos Biomecânicos/fisiologia , Reabsorção Óssea/diagnóstico por imagem , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/cirurgia , Fusão Vertebral/instrumentação , Estenose Espinal/diagnóstico por imagem , Adulto , Remodelação Óssea/fisiologia , Reabsorção Óssea/fisiopatologia , Feminino , Análise de Elementos Finitos , Humanos , Disco Intervertebral/fisiologia , Vértebras Lombares/fisiopatologia , Imagem por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Amplitude de Movimento Articular/fisiologia , Fusão Vertebral/efeitos adversos , Estenose Espinal/cirurgia , Tomografia Computadorizada por Raios X , Resultado do Tratamento
9.
World Neurosurg ; 113: e70-e76, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29408574

RESUMO

OBJECTIVE: To assess the mid-long-term follow-up of the safety and efficacy of anterior cervical discectomy and fusion (ACDF), cervical artificial disc replacement (CADR) and hybrid surgery (HS) for bilevel cervical degenerative disc disease (cDDD). METHODS: 77 patients who underwent ACDF, HS, and CADR were retrospectively reviewed. Clinical effects were evaluated based on Neck Disability Index (NDI), Visual Analog Scale (VAS), and Japanese Orthopedic Association (JOA) scores and the Odom criteria. Radiographic outcomes were evaluated, including cervical range of motion (ROM), ROM in the operative and adjacent segments, incidence of degeneration in the adjacent segments (ASD), and heterotopic ossification (HO). RESULTS: NDI, VAS, and JOA scores significantly improved in all patients after surgery without significant differences between groups. The excellent-to-good ratio in the Odom scale was 28/30 for the HS group, 30/33 for the ACDF group, and 13/14 for the CADR group. No significant differences in clinical outcomes or complication were found between groups (P > 0.05). Furthermore, the HS and CADR groups had less decreased ROM in the cervical and operative segments and less compensatory ROM in adjacent segments (P < 0.05). By contrast, the ACDF group had decreased ROM in the cervical and operative segments and significantly increased ROM in adjacent segments (P < 0.05). Moreover, the incidence of ASD was higher in the ACDF group, but the difference was not statistically significant (P > 0.05). HO was found in 10 patients (33.3%) in the HS group and 5 patients (35.7%) in the CADR group. CONCLUSION: HS was superior to ACDF with regard to equivalent clinical outcomes in the mid-long-term follow-up. Furthermore, HS was superior in the maintenance of ROM and had less impact on its adjacent segments. The efficacy of HS is similar to that of CADR.


Assuntos
Vértebras Cervicais/cirurgia , Discotomia/métodos , Degeneração do Disco Intervertebral/cirurgia , Fusão Vertebral/métodos , Substituição Total de Disco/métodos , Idoso , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/patologia , Progressão da Doença , Feminino , Seguimentos , Humanos , Degeneração do Disco Intervertebral/complicações , Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/patologia , Masculino , Pessoa de Meia-Idade , Cervicalgia/etiologia , Ossificação Heterotópica/epidemiologia , Ossificação Heterotópica/etiologia , Medição da Dor , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Próteses e Implantes , Amplitude de Movimento Articular , Estudos Retrospectivos , Índice de Gravidade de Doença , Resultado do Tratamento , Escala Visual Analógica
10.
Chin Med J (Engl) ; 130(21): 2535-2540, 2017 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-29067951

RESUMO

BACKGROUND: Thoracolumbar junction (TLJ) is the transitional area between the lower thoracic spine and the upper lumbar spine. Vertebral compression fractures and proximal junctional kyphosis following spine surgery often occur in this area. Therefore, the study of development and mechanisms of thoracolumbar junctional degeneration is important for planning surgical management. This study aimed to review radiological parameters of thoracolumbar junctional degenerative kyphosis (TLJDK) in patients with lumbar degenerative kyphosis and to analyze compensatory mechanisms of sagittal balance. METHODS: From January 2016 to March 2017, patients with lumbar degenerative kyphosis were enrolled in this radiographic study. Patients were divided into two groups according to thoracolumbar junctional angle (TLJA): the non-TLJDK (NTLJDK) group (TLJA <10°) and the TLJDK group (TLJA ≥10°). Complete spinopelvic radiographic parameters were analyzed and compared between two groups. Pearson or Spearman correlation coefficients and independent two-sample t-test or Mann-Whitney U-test were used. RESULTS: A total of 77 patients with symptomatic sagittal imbalance due to lumbar degenerative kyphosis were enrolled in this study. There were 34 patients in NTLJDK group (TLJA <10°) and 43 patients in TLJDK group (TLJA ≥10°). The median angle of lumbar lordosis (LL) in the NTLJDK or TLJDK groups was 23.40° (18.50°, 29.48°) or 19.50° (13.30°, 24.55°), respectively. The median TLJAs in all patients and both groups were -11.20° (-14.60°, -4.80°), -3.70° (-7.53°, -1.73°), and -14.30° (-17.45°, -13.00°), respectively. In the NTLJDK group, LL was correlated with thoracic kyphosis (TK; r = -0.400, P = 0.019), sacral slope (SS; r = 0.681, P < 0.001), and C7-sagittal vertical axis (r = -0.402, P = 0.018). In the TLJDK group, LL was correlated with TK (r = -0.345, P = 0.024), SS (r = 0.595, P < 0.001), and pelvic tilt (r = -0.363, P = 0.017). There were significant differences in LL, TLJA, TK, SS, and pelvic incidence (PI) between two groups. CONCLUSIONS: Although TLJDK is common in patients with lumbar degenerative kyphosis, it might be generated by special characteristics of morphology and biomechanics of the TLJ. To maintain sagittal balance, pelvis back tilt might be more important in patients with TLJDK, whereas thoracic curve changes might be more important in patients without TLJDK.


Assuntos
Cifose/diagnóstico por imagem , Fraturas da Coluna Vertebral/diagnóstico por imagem , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Cifose/patologia , Lordose/diagnóstico por imagem , Lordose/patologia , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/patologia , Masculino , Pessoa de Meia-Idade , Escoliose/diagnóstico por imagem , Escoliose/patologia , Fraturas da Coluna Vertebral/patologia , Vértebras Torácicas/diagnóstico por imagem , Vértebras Torácicas/patologia
11.
Oncotarget ; 7(48): 78985-78993, 2016 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-27738317

RESUMO

OBJECTIVES: Estimate the epidermal growth factor receptor (EGFR) mutation prevalence in all non-small cell lung cancer (NSCLC) patients and patient subgroups. RESULTS: A total of 456 studies were included, reporting 30,466 patients with EGFR mutation among 115,815 NSCLC patients. The overall pooled prevalence for EGFR mutations was 32.3% (95% CI 30.9% to 33.7%), ranging from 38.4% (95% CI: 36.5% to 40.3%) in China to 14.1% (95% CI: 12.7% to 15.5%) in Europe. The pooled prevalence of EGFR mutation was higher in females (females vs. males: 43.7% vs. 24.0%; OR: 2.7, 95% CI: 2.5 to 2.9), non-smokers (non-smokers vs. past or current smokers: 49.3% vs. 21.5%; OR: 3.7, 95% CI: 3.4 to 4.0), and patients with adenocarcinoma (adenocarcinoma vs. non-adenocarcinoma: 38.0% vs. 11.7%; OR: 4.1, 95% CI: 3.6 to 4.8). MATERIALS AND METHODS: PubMed, EMBASE, and the Cochrane Library were searched to June 2013. Eligible studies reported EGFR mutation prevalence and the association with at least one of the following factors: gender, smoking status and histology. Random-effects models were used to pool EGFR mutation prevalence data. CONCLUSION: This study provides the exact prevalence of EGFR mutations in different countries and NSCLC patient subgroups.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Receptores ErbB/genética , Neoplasias Pulmonares/genética , Mutação , Grupo com Ancestrais do Continente Asiático/genética , China , Europa (Continente) , Grupo com Ancestrais do Continente Europeu/genética , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Taxa de Mutação , Prevalência , Fatores de Risco , Fumar/genética
12.
Biomed Environ Sci ; 27(5): 391-5, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24827722

RESUMO

The effect of sterilization methods on biological activity of fibronectin on the surface of biomaterials was elaborated in the present study. Sterile protein- modified biomaterials were fabricated by microfilter filtration and UV irradiation, respectively. UV irradiation altered the conformation of surface- adsorbed fibronectin and further affected the attachment, morphology and biological function of endothelial cells. However, microfilter filtration did not to change the normal conformation of fibronectin, or the proliferation and biological function of endothelial cells, indicating that microfilter filtration sterilization is the most suitable method for protein-substrate.


Assuntos
Fibronectinas/efeitos da radiação , Próteses e Implantes/microbiologia , Esterilização/métodos , Adesão Celular/efeitos da radiação , Filtração , Raios Ultravioleta
13.
Huan Jing Ke Xue ; 35(10): 3794-9, 2014 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-25693385

RESUMO

This paper studied the phtodegradation behavior, mechanism and security of propranolol (PRO) in water under ultraviolet irradiation using the high pressure mercury lamp. The photodegradation mechanism was verified by reactive oxygen species (ROS) quenching method, and the photoproducts security was evaluated by luminescent bacteria toxicity test. The results showed that the rate of photolysis (k) of PRO decreased with increasing initial concentration, and showed a significant negative correlation (r2 > 0.95). The increasing k value was also accompanied with higher pH of the solution, and the k values were 0.0953-0.267 min(-1) under pH 5-9. ROS quenching experiments showed that the PRO UV photolysis process included a triplet PRO (3PRO*) direct photolysis participation, and self-sensitized photolysis participation caused by hydroxyl radical (·OH) and singlet oxygen (1O2). Direct photolysis rate was greater than the rate of self-sensitized photolysis. The result of 1O2 steady concentration measured by FFA as the probe was consistent with the quenching method. The toxicity evaluation illustrated the formation of some intermediate photoproducts, which were more toxic than PRO.


Assuntos
Fotólise , Propranolol/química , Raios Ultravioleta , Bactérias , Radical Hidroxila/química , Medições Luminescentes , Propranolol/efeitos da radiação , Oxigênio Singlete/química , Soluções , Testes de Toxicidade , Água
14.
Asian Pac J Cancer Prev ; 14(10): 6151-8, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24289641

RESUMO

Radio frequency ablation (RFA) is an effective means of achieving local control of liver cancer. It is a particularly suitable mode of therapy for small and favorably located tumors. However, local progression rates are substantially higher for large tumors (>3.0 cm). In the current study, we report on a mathematical model based on geometric optimization to treat large liver tumors. A database of mathematical models relevant to the configuration of liver cancer was also established. The specific placement of electrodes and the frequency of ablation were also optimized. In addition, three types of liver cancer lesion were simulated by computer guidance incorporating mathematical models. This approach can be expected to provide a more effective and rationale mechanism for employing RFA in the therapy of hepatic carcinoma.


Assuntos
Ablação por Cateter , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/cirurgia , Modelos Teóricos , Adulto , Idoso , Simulação por Computador , Eletrodos , Feminino , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Prognóstico , Tomografia Computadorizada por Raios X , Adulto Jovem
15.
Zhonghua Wai Ke Za Zhi ; 51(5): 426-31, 2013 May 01.
Artigo em Chinês | MEDLINE | ID: mdl-23958166

RESUMO

OBJECTIVE: To investigate the effects and apoptosis of intrathecal injection of Methylprednisolone Sodium Succinate (MPss) for acute spinal cord injury (SCI) in New Zealand rabbits. METHODS: Seventy-two healthy New Zealand rabbits were used for the procedure and were randomly divided into two groups: SCI group and SHAM group, which was both divided into 6 subgroups, such as the vehicle group, the MPss intrathecal injection groups (1.5 mg/kg, 3.0 mg/kg, 6.0 mg/kg group), the MPss intravenous injection group and the combined injection group. TARLOV score was tested daily to evaluate the motor function. The rabbits were sacrificed 7 days after the surgery and the thoracic spinal cord sections and the sacral sections where MPss was injected were harvested for HE and TUNEL staining. Two-Factors Repeated Measures analysis of variance for TARLOV scores tested at various times and One-Way ANOVA analysis of variance for data between groups were used. RESULT: Seven days after surgery in SCI group, there was no statistical difference between the TARLOV scores of intrathecal injection of MPss 3.0 mg/kg group, 6.0 mg/kg group and MPss intravenous injection group (P > 0.05), which were all better than the vehicle group (F = 4.762, P < 0.05). Referring to the lymphocyte infiltration at the injury site in SCI group, there was statistical difference between MPss intrathecal injection 6.0 mg/kg group (1.33 ± 0.21) and the vehicle group (2.67 ± 0.21) (F = 5.793, P < 0.05) and no statistical difference between intrathecal injection of MPss 6.0 mg/kg group and MPss intravenous injection group (P > 0.05). As for the lymphocyte infiltration at the intrathecal injection site in SHAM group, there was statistical difference between MPss intrathecal injection 6.0 mg/kg group (2.50 ± 0.55) and the vehicle group (0.50 ± 0.55) (F = 17.333, P < 0.05). TUNEL staining in SCI group showed statistical difference between MPss intrathecal injection 6.0 mg/kg group (6.3 ± 1.5) and the vehicle group (20.3 ± 2.2) (F = 71.279, P < 0.05). CONCLUSIONS: Intrathecal injection of MPss can improve the functional recovery of lower limb and decrease apoptosis of neuron cells,which can provide same effects as the traditional intravenous injection of MPss in New Zealand rabbits.


Assuntos
Hemissuccinato de Metilprednisolona/uso terapêutico , Traumatismos da Medula Espinal/tratamento farmacológico , Doença Aguda , Análise de Variância , Animais , Modelos Animais de Doenças , Injeções Espinhais , Masculino , Hemissuccinato de Metilprednisolona/administração & dosagem , Coelhos , Recuperação de Função Fisiológica
16.
PLoS One ; 6(12): e28491, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22164300

RESUMO

Anthracycline daunorubicin (DNR) is one of the major antitumor agents widely used in the treatment of myeloid leukemia. Unfortunately, the clinical efficacy of DNR was limited because of its cytotoxity at high dosage. As a novel cytoprotective mechanism for tumor cell to survive under unfavorable conditions, autophagy has been proposed to play a role in drug resistance of tumor cells. Whether DNR can activate to impair the sensitivity of cancer cells remains unknown. Here, we first report that DNR can induce a high level of autophagy, which was associated with the activation of extracellular signal-regulated kinase 1/2 (ERK1/2). Moreover, cell death induced by DNR was greatly enhanced after autophagy inhibition by the pharmacological inhibitor chloroquine (CQ) and siRNAs targeting Atg5 and Atg7, the most important components for the formation of autophagosome. In conclusion, we found that DNR can induce cytoprotective autophagy by activation of ERK in myeloid leukemia cells. Autophagy inhibition thus represents a promising approach to improve the efficacy of DNR in the treatment of patients with myeloid leukemia.


Assuntos
Apoptose , Autofagia , Daunorrubicina/farmacologia , Antibióticos Antineoplásicos/farmacologia , Antirreumáticos/farmacologia , Cloroquina/farmacologia , Sinergismo Farmacológico , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Humanos , Células K562 , Leucemia Mieloide/metabolismo , Microscopia Confocal/métodos , Microscopia Eletrônica de Transmissão/métodos , Neoplasias/metabolismo , RNA Interferente Pequeno/metabolismo , Transdução de Sinais
17.
Huan Jing Ke Xue ; 31(9): 2184-90, 2010 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-21072944

RESUMO

The bioflocculant ZS-7 was purified to homogeneity by ethanol precipitation, dialysis and gel permeation chromatography (GPC). About 0.98 g of the purified bioflocculant could be recovered from 1L of fermentation broth. The purified bioflocculant was identified as a glycoprotein consisting of polysaccharide (91.5%) and protein (8.4%), with an approximate molecular weight of 6.89 x 10(4). The major component of ZS-7 is an acid polysaccharide including uronic (16.4%), pyruvic (7.1%) and acetic acids (0.5%). It consists of galactose, glucose, mannose and rhamnose in a molar ratio of 142 :2.2 : 4.5 : 3.4. X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) and NMR spectrum of the bioflocculant indicate the presence of carboxyl, hydroxyl, amide, amino, methoxyl and sulfate groups.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Glicoproteínas/química , Glicoproteínas/isolamento & purificação , Manose/análise , Ramnose/análise , Floculação , Galactose/análise , Peso Molecular , Polissacarídeos/análise , Proteínas/análise
18.
Hepatogastroenterology ; 57(102-103): 1220-6, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21410062

RESUMO

BACKGROUND/AIMS: The metastasis of hepatic carcinoma is correlated with the body's immune status. T lymphocytes play a big part role in tumor immune. The aim of the present study is to investigate the inhibition effects of metastatsis in nude mice bearing hepatic carcinoma after T lymphocytes reconstitution. METHODOLOGY: An immune reconstitution model was established in nude mice. Then, 42 nude mice were distributed into 4 groups for T lymphocytes reconstitution. The lymph nodes of each group were obtained to investigate the tumor metastasis. And the secretion of cytokines and the apoptosis of tumor cells in each group were also detected. RESULTS: The ratio of CD3, CD4, CD8, CD4/CD8 in reconstituted groups were higher than controlled groups. The average time of tumor formation in Balb/c nu/nu mice was 7.7 +/- 0.6 days and in Balb/c mice was 11.5 +/- 1.3 days. After active T lymphocytes reconstitution, the extent lymph nodes metastasis in reconstitution groups was lower than control groups (p < 0.05), but proximal metastasis has no significant difference. The level of serum IL-10 in nude mice after immune reconstitution was significantly lower and VEGF was significantly higher than that of the control group (p < 0.05). Apoptosis of the hepatic carcinoma cells was increased significantly after immune reconstitution (p < 0.05). CONCLUSIONS: The metastasis of hepatic carcinoma can be inhibited by reconstitution of actived T lymphocytes in nude mice, which indicated that tumor metastasis can be affected by the immune status in host body.


Assuntos
Carcinoma Hepatocelular/secundário , Neoplasias Hepáticas/patologia , Ativação Linfocitária , Linfócitos T/imunologia , Animais , Carcinoma Hepatocelular/patologia , Citocinas/biossíntese , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Metástase Neoplásica/prevenção & controle
19.
Zhonghua Gan Zang Bing Za Zhi ; 16(1): 12-6, 2008 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-18226336

RESUMO

OBJECTIVE: We previously showed that introduction of a normal, neomycin-tagged human chromosome 8 reduced the metastatic capacity of C5F rat liver cancer cell line, which had high metastatic potential without affecting tumorigenicity, suggesting the presence of one or more metastasis suppressor genes encoded on human chromosome 8. We proceeded to define further the region harboring the metastasis suppressor gene(s) and to determine the random loss of human chromosome 8 by PCR amplification of sequence tag site (STS) markers. METHODS: The national Center for Biotechnology Information (NCBI) databases were used as references of the relative genetic distances of the STS markers. C5F genomic DNA and A9/neo8 genomic DNA were used as negative and positive controls for chromosome 8 amplification, respectively. Genomic DNA was isolated and quantified from cultured hybrid clones (A9/C5F-1 and A9/C5F-2 microcell hybrid clones served as metastasis-unsuppressed groups; A9/C5F-4, A9/C5F-8 and A9/C5F-10 microcell hybrid clones served as metastasis suppressed groups). STS-PCR products were separated by electrophoresis through 2% agarose gel. RESULTS: Metastasis-suppressed microcell hybrid clones (A9/C5F-4, A9/C5F-8 and A9/C5F-10) conserved STS markers between D8S542 --> D8S1973 (8p21.1-23.1). In contrast, metastasis-unsuppressed clones (A9/C5F-1 and A9/C5F-2) lacked several markers in this region. In attempts to refine the region retained in the microcell suppressed clones, more densely spaced STS markers in the human chromosome 8p21.1-23.1 were used. We found that the metastasis-suppressed clones retained 18cM region between D8S542 and D8S1973 (8P21.1-23.1), where as the metastasis-unsuppressed clones lacked the region. CONCLUSION: Our results suggest that a metastasis suppressor gene is located within the interval between D8S542 and D8S1973 on human chromosome 8p21.1-23.1.


Assuntos
Carcinoma Hepatocelular/genética , Cromossomos Humanos Par 8/genética , Genes Supressores de Tumor , Neoplasias Hepáticas/genética , Sitios de Sequências Rotuladas , Linhagem Celular , Linhagem Celular Tumoral , Mapeamento Cromossômico , Fibroblastos/citologia , Humanos , Hibridização in Situ Fluorescente , Metástase Neoplásica
20.
Zhonghua Yi Xue Za Zhi ; 87(29): 2050-3, 2007 Aug 07.
Artigo em Chinês | MEDLINE | ID: mdl-17925177

RESUMO

OBJECTIVE: To investigate the relation between the changes of dendritic cell (DC) function and down-regulation of beta-centractin in hepatocellular carcinoma. METHODS: DC derived from peripheral blood were cultured and then pulsed by lysates from hepatocarcinoma cells (HCC) with high, low, or none metastatic potential of the lines HCCLM6, MHCC97L, and Hep3B, and from normal human liver cell of the line Chang liver. DC not pulsed were used as control group. Three days later scanning electron microscopy and inverted microscopy were used to observe the morphology of the DC. Flow cytometry was used to observe the phenotype. The protein expression of beta-centractin was detected by Western blotting and immunocytochemistry. RESULTS: Scanning electron microscopy and inverted microscopy showed no change in the morphology of the DC pulsed by different antigens. The expression levels of HLA-DR, CD80, CD83, and CD86 of the 4 pulsed groups were all significantly higher than that of the un-pulsed group (all P < 0.05). The expression of CD86 of the DC + LM6 group was significantly lower than those of the DC + Chang, DC + Hep3B, and DC + 97L groups (all P < 0.05). The mixed lymphocyte reaction (MLR) levels of the 4 pulsed groups were all significantly higher than that of the control group (all P < 0.05). The MLR level of the DC + LM6 group was significantly lower than those of the DC + Chang, DC + Hep3B, and DC + 97L groups (all P < 0.05). Western blotting showed that beta-centractin was not expressed in the control DC and was expressed in the 3 pulsed DC groups, and the beta-centractin expression levels of the DC + Chang, DC + Hep3B, and DC + 97L groups were all higher than that of the DC + LM6 group. The results of immunocytochemistry were similar to that of Western blotting. CONCLUSION: The down-regulation of beta-centractin in the DC pulsed with high metastatic potential HVV cell lysates is associated DC dysfunction and may be one of the mechanisms of HCC immune escape.


Assuntos
Actinas/biossíntese , Extratos Celulares/farmacologia , Células Dendríticas/efeitos dos fármacos , Western Blotting , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular , Linhagem Celular Tumoral , Células Dendríticas/citologia , Células Dendríticas/metabolismo , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia
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