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1.
Molecules ; 29(9)2024 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-38731462

RESUMO

A novel and efficient method for functionalizing organosulfones has been established, utilizing a visible-light-driven intermolecular radical cascade cyclization of α-allyl-ß-ketosulfones. This process employs fac-Ir(ppy)3 as the photoredox catalyst and α-carbonyl alkyl bromide as the oxidizing agent. Via this approach, the substrates experience intermolecular addition of α-carbonyl alkyl radicals to the alkene bonds, initiating a sequence of C-C bond formations that culminate in the production of organosulfone derivatives. Notably, this technique features gentle reaction conditions and an exceptional compatibility with a wide array of functional groups, making it a versatile and valuable addition to the field of organic synthesis.

2.
Plant Physiol ; 189(2): 1021-1036, 2022 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-35234946

RESUMO

Soybean (Glycine max) is highly sensitive to photoperiod, which affects flowering time and plant architecture and thus limits the distribution range of elite soybean cultivars. The major maturity gene E1 confers the most prominent effect on photoperiod sensitivity, but its downstream signaling pathway remains largely unknown. Here, we confirm that the encoded E1 protein is a transcriptional repressor. The expression of seven GmMDE genes (Glycine max MADS-box genes downregulated by E1) was suppressed when E1 was overexpressed and promoted when E1 was knocked out through clustered regularly-interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-mediated mutagenesis. These GmMDEs exhibited similar tissue specificity and expression patterns, including in response to photoperiod, E1 expression, and E1 genotype. E1 repressed GmMDE promoter activity. Results for two GmMDEs showed that E1 epigenetically silences their expression by directly binding to their promoters to increase H3K27me3 levels. The overexpression of GmMDE06 promoted flowering and post-flowering termination of stem growth. The late flowering phenotype of E1-overexpressing soybean lines was reversed by the overexpression of GmMDE06, placing GmMDE06 downstream of E1. The overexpression of GmMDE06 increased the expression of the soybean FLOWERING LOCUS T orthologs GmFT2a and GmFT5a, leading to feedback upregulation of GmMDE, indicating that GmMDE and GmFT2a/GmFT5a form a positive regulatory feedback loop promoting flowering. GmMDE06 also promoted post-flowering termination of stem growth by repressing the expression of the shoot identity gene Dt1. The E1-GmMDEs-GmFT2a/5a-Dt1 signaling pathway illustrates how soybean responds to photoperiod by modulating flowering time and post-flowering stem termination.


Assuntos
Glycine max , Fotoperíodo , Florígeno/metabolismo , Flores/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Glycine max/metabolismo
3.
World J Gastrointest Surg ; 16(1): 215-227, 2024 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-38328319

RESUMO

BACKGROUND: Postoperative complications remain a paramount concern for surgeons and healthcare practitioners. AIM: To present a comprehensive analysis of the Estimation of Physiologic Ability and Surgical Stress (E-PASS) scoring system's efficacy in predicting postoperative complications following abdominal surgery. METHODS: A systematic search of published studies was conducted, yielding 17 studies with pertinent data. Parameters such as preoperative risk score (PRS), surgical stress score (SSS), comprehensive risk score (CRS), postoperative complications, postoperative mortality, and other clinical data were collected for meta-analysis. Forest plots were employed for continuous and binary variables, with χ2 tests assessing heterogeneity (P value). RESULTS: Patients experiencing complications after abdominal surgery exhibited significantly higher E-PASS scores compared to those without complications [mean difference and 95% confidence interval (CI) of PRS: 0.10 (0.05-0.15); SSS: 0.04 (0.001-0.08); CRS: 0.19 (0.07-0.31)]. Following the exclusion of low-quality studies, results remained valid with no discernible heterogeneity. Subgroup analysis indicated that variations in sample size and age may contribute to heterogeneity in CRS analysis. Binary variable meta-analysis demonstrated a correlation between high CRS and increased postoperative complication rates [odds ratio (OR) (95%CI): 3.01 (1.83-4.95)], with a significant association observed between high CRS and postoperative mortality [OR (95%CI): 15.49 (3.75-64.01)]. CONCLUSION: In summary, postoperative complications in abdominal surgery, as assessed by the E-PASS scoring system, are consistently linked to elevated PRS, SSS, and CRS scores. High CRS scores emerge as risk factors for heightened morbidity and mortality. This study establishes the accuracy of the E-PASS scoring system in predicting postoperative morbidity and mortality in abdominal surgery, underscoring its potential for widespread adoption in effective risk assessment.

4.
Genes Cells ; 17(3): 205-17, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22251225

RESUMO

Multidrug resistance (MDR) is a major problem facing patients with cancer. Although Neutrophil gelatinase-associated lipocalin (NGAL) is highly expressed in various cancers, the possible role of NGAL in MDR is still obscure. In this article, we evaluated the effect of NGAL on Rh123 accumulation in cancer cells. NGAL was first down-regulated by short hairpin RNA-mediated interference. In correlation with the reduced NGAL expression, intracellular Rh123 accumulation was significantly decreased. We finally observed that inhibiting both of the ERK1/2 and p38 MAPK could seriously down-regulate NGAL expression and also decrease the intracellular accumulation of Rh123, indicating that NGAL-mediated Rh123 accumulation is regulated by the phosphorylation of ERK1/2 and p38 MAPK. Pretreatment of MDA-MB-231 with NGAL recombinant protein and antibody had significant effects on the intracellular accumulation of Rh123, whereas little effect was observed in K562 cells treated with the same method, suggesting that NGAL was involved in the regulation of Rh123 accumulation in these two types of cancers, although different pathways. Here we provide new evidence that directly shows the possibility of small chemical substances Rh123 intracellular accumulation that is regulated by NGAL. These results suggest the possibility of NGAL involvement in drug transportation and cancer MDR formation, and indicate the potential of NGAL in cancer therapy.


Assuntos
Proteínas de Fase Aguda/metabolismo , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Corantes Fluorescentes/metabolismo , Lipocalinas/metabolismo , Neoplasias/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Rodamina 123/metabolismo , Proteínas de Fase Aguda/genética , Transporte Biológico , Linhagem Celular Tumoral , Técnicas de Silenciamento de Genes , Humanos , Lipocalina-2 , Lipocalinas/genética , Proteínas Proto-Oncogênicas/genética
5.
Ultrastruct Pathol ; 37(2): 93-101, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23573889

RESUMO

OBJECTIVE: To describe characteristics of monocytes and histiocytes in the bone marrow of patients with a confirmed and suspected diagnosis of reactive histiocytosis. METHODS: 14 patients with a confident diagnosis of reactive histiocytosis or with a suspected diagnosis were inpatients at the Tianjin Blood Diseases Hospital between 2008 and 2012. Nucleated cells from bone marrow were observed by light microscopy - morphologically and immunohistochemically for histiocyte antigens - and ultrastructurally by transmission electron microscopy. RESULTS: Monocytes, atypical histiocytes, macrophages, hemophagocytes, reticular cells and dendritic cells were significantly increased in 9, 9, 5, 3, 3 and 2, respectively, of the 14 cases. Atypical histiocytes expressed some morphological characteristics of promonocytes. CONCLUSION: Monocytes, atypical histiocytes, macrophages, hemophagocytes, reticular cells and dendritic cells were increased in different relative degrees in patients with bone marrow reactive histiocytosis or suspected reactive histiocytosis. The increase in numbers of monocytes, atypical histiocytes and macrophages was a particularly significant feature. It is argued that atypical histiocytes with immature monocyte features might be precursors of hemophagocytes, reticular cells or dendritic cells.


Assuntos
Células da Medula Óssea/ultraestrutura , Medula Óssea/ultraestrutura , Histiócitos/ultraestrutura , Histiocitose de Células não Langerhans/patologia , Monócitos/ultraestrutura , Adolescente , Adulto , Idoso , Antígenos de Diferenciação/metabolismo , Medula Óssea/metabolismo , Células da Medula Óssea/metabolismo , Exame de Medula Óssea , Contagem de Células , Pré-Escolar , Células Dendríticas/metabolismo , Células Dendríticas/ultraestrutura , Feminino , Fibroblastos/metabolismo , Fibroblastos/ultraestrutura , Histiócitos/metabolismo , Humanos , Lactente , Masculino , Microscopia Eletrônica de Transmissão , Monócitos/metabolismo , Fagócitos/metabolismo , Fagócitos/ultraestrutura , Reticulócitos/metabolismo , Reticulócitos/ultraestrutura , Adulto Jovem
6.
J Biophotonics ; 16(11): e202300190, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37545092

RESUMO

The dichroic mirror (DM) is a key component in microscope. We found a ghost in the reflection channel of a dual-channel fluorescence microscope and studied the relationship between the ghost and the incidence angle θ into the DM. The DM emission surface reflection generated ghost if the θ is not 45 ° . We analyzed the distance and intensity relationship between the ghost and the primary image, which is θ -dependent and was demonstrated by imaging live cells and a stage micrometer. The ghost can be eliminated by placing the DM between objective and tube lens, but not between tube lens and detector, ensuring that the incident light into the DM is approximately parallel. Furthermore, the transmitted light of the DM is shifted towards a longer wavelength with increasing θ . Collectively, microscopists must carefully optimize the θ when designing a microscope to avoid the ghost.


Assuntos
Microscopia de Fluorescência , Microscopia de Fluorescência/métodos
7.
Se Pu ; 41(10): 911-920, 2023 Oct.
Artigo em Zh | MEDLINE | ID: mdl-37875413

RESUMO

Neurotransmitters (NTs) are essential for intercellular communication and primarily include monoamine, amino acid, and cholinergic NTs. These molecules play important roles in the body's stress response, motor coordination, neuronal communication, and homeostatic functions. Previous studies have shown that abnormal changes in NT levels are associated with various neurological disorders. Therefore, the development of accurate analytical methods for NT detection will enhance the current understanding on complex neuropathophysiology by providing functional knowledge and techniques for early diagnosis, thereby facilitating the development of new therapeutic options for the related diseases. The solid phase microextraction (SPME) technique combines sample preparation, separation, and enrichment in a single step and is minimally invasive, low cost, solvent free, and high throughput. SPME has been successfully applied to the in vivo analysis of target analytes in animal, human, and plant tissues. The coating material plays a significant role in the development of in vivo SPME methods and must meet various analytical requirements, including a suitable geometry for the SPME device, high extraction capacity, excellent selectivity, and wide extraction coverage for the target analytes. Covalent organic frameworks (COFs) are porous crystalline polymers constructed from organic framework units through strong covalent bonds; these materials are characterized with a low density, large specific surface area, permanent porosity, excellent chemical/thermal stability, and easy functionalization.In this study, a sulfonic acid-functionalized COF material (COF-SO3H) with good crystallinity, excellent chemical/thermal stability, strong hydrophobicity, a uniform mesoporous structure, and narrow pore size distribution was prepared using 2,4,6-triformylphloroglucinol and 1,4-diamino-2-nitrobenzene as monomers. Then, the COF-SO3H was coated onto the surface of stainless-steel fibers and used for in vivo enrichment of NTs. The structural properties of COF-SO3H were characterized using various techniques, such as scanning electron microscopy (SEM), Fourier transform-infrared spectroscopy (FT-IR), and X-ray diffraction (XRD), all of which showed that COF-SO3H had a good crystalline structure and uniform mesopore distribution with a specific surface area of 46.17 m2/g. Compared with the SPME fibers of HLB, C18, MCX, amino, and PXC columns, the prepared COF-SO3H fibers showed better extraction efficiency for the target NTs. Next, the factors affecting SPME efficiency were optimized. The optimal desorption solvent was formic acid-methanol-water (0.5∶49.5∶50, v/v/v), and the optimal extraction and desorption times were 15 min. A method for the in vivo analysis of NTs in the brains of mice was established by combining the COF-SO3H fibers with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) under optimal conditions. The NTs were separated on an Acquity UPLC BEH-C18 analytical column (100 mm×2.1 mm, 1.7 µm) with 0.1% formic acid aqueous solution (A) and acetonitrile (B) as the mobile phases. The flow rate was set to 0.2 mL/min, and the gradient elution procedure was as follows: 0-4 min, 5%B-6%B; 4-7 min, 6%B-5%B; 7-11 min, 5%B. Under optimal conditions, the method showed good linearity (r2>0.99). The limits of quantification (S/N≥5) were in the range of 0.003-0.005 µg/mL and 3-5 µg/mL for monoamine and amino acid NTs, respectively, with RSDs of less than 20%. The method showed good precision (0.80%-9.70%) and accuracy (2.08%-17.72%), with absolute matrix effects in the range of 82.22%-117.92%. These values reflect the good purification and enrichment abilities of the proposed fibers for the target analytes. Finally, the established SPME method was combined with UPLC-MS/MS and successfully applied to quantify target NTs in the brains of mice. The proposed strategy provides a practical method for the in vivo detection and quantitative analysis of NTs and expands the applications of functionalized COF materials for the analysis of various targets.


Assuntos
Estruturas Metalorgânicas , Humanos , Animais , Camundongos , Cromatografia Líquida , Microextração em Fase Sólida , Espectroscopia de Infravermelho com Transformada de Fourier , Espectrometria de Massas em Tandem , Aminas , Aminoácidos , Encéfalo , Neurotransmissores , Extração em Fase Sólida , Cromatografia Líquida de Alta Pressão
8.
Genes Cells ; 16(4): 416-26, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21392185

RESUMO

The interaction between calcineurin B homologous protein 2 (CHP2) and Na(+) /H(+) exchanger 1 (NHE1), two membrane proteins, is essential for protecting cells from serum deprivation-induced death. Although four putative EF-hands in CHP2 had been predicted for years, Ca²(+) -binding activities of these motifs have not been tested yet, their role in this process remain poorly understood. To identify Ca²(+) -binding motifs required for the stable formation of CHP2/NHE1 complexes, we developed a mutagenesis-based assay in PS120 cells. We found that (45) Ca²(+) bond to two EF-hand motifs (EF3 and 4) of CHP2 proteins with high affinity. Complex formation between CHP2 and the CHP2 binding domain of NHE1 resulted in a marked increase in the Ca²(+) -binding affinity of CHP2. Co-immunoprecipitation and distribution of GFP-tagged CHP2-EF3m/4m also indicated that Ca²(+) affected the membrane location of CHP2 to interact with NHE1. The C-terminal region of CHP2 contains a nuclear export sequence (NES). When the six leucines of NES were mutated to alanines, the resulting CHP2 protein was predominantly localized to the nucleus. Furthermore, mutation of the NES resulted in enhanced proliferation and oncogenic potential of HeLa cells. Together, these results show that calcium and NES control the subcellular distribution of CHP2 and then distinctively regulate cell proliferation.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Proliferação de Células , Motivos EF Hand/genética , Sinais de Localização Nuclear/química , Trocadores de Sódio-Hidrogênio/metabolismo , Sequência de Aminoácidos , Proteínas de Ligação ao Cálcio/química , Linhagem Celular Transformada , Células HeLa/metabolismo , Humanos , Dados de Sequência Molecular , Sinais de Localização Nuclear/fisiologia , Alinhamento de Sequência , Trocador 1 de Sódio-Hidrogênio
9.
Front Plant Sci ; 13: 992755, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36352884

RESUMO

Drought is the abiotic factor that adversely affects plant growth, development survival, and crop productivity, posing a substantial threat to sustainable agriculture worldwide, especially in warm and dry areas. However, the extent of damage depends upon the crop growth stage, severity and frequency of the stress. In general, the reproductive growth phase is more sensitive to stresses causing a substantial loss in crop productivity. Saccharum spontaneum (L.) is the most variable wild relative of sugarcane with potential for use in sugarcane crop improvement programs. In the present study addresses the transcriptomic analysis of drought stress imposed by polyethylene glycol-6000 (PED-6000; w/v- 25%) on the root tip tissues of S. spontaneum GX83-10. The analysis of microarrays of drought-stressed roots was performed at 0 (CK), 2 (T2), 4 (T4), 8 (T8) and 24 h (T24). The analyzed data were compared with the gene function annotations of four major databases, such as Nr, KOG/COG, Swiss-Prot, and KEGG, and a total of 62,988 single-gene information was obtained. The differently expressed genes of 56237 (T4), 59319 (T8), and 58583 (T24), among which CK obtained the most significant number of expressed genes (35920) as compared to T24, with a total of 53683 trend genes. Gene ontology (GO) and KEGG analysis were performed on the 6 important trends, and a total of 598 significant GO IDs and 42 significantly enriched metabolic pathways. Furthermore, these findings also aid in the selection of novel genes and promoters that can be used to potentially produce crop plants with enhanced stress resistance efficiency for sustainable agriculture.

10.
Front Plant Sci ; 13: 852886, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35371161

RESUMO

Sugarcane is a cash crop that plays an integral part in the sugar industry. The Sustainable Sugarcane Initiative (SSI) has been adopted globally, ensuring enough and aiming for more yield, helping increase disease-free sugarcane cultivation. Single-bud seeds could be the best approach for sugarcane cultivation. Indole-3-butyric acid (IBA) is a rooting agent utilized significantly in seedling propagation. Greenhouse experiment results discovered the significant growth promotion in sugarcane seedlings and accumulation of plant hormones at 100 ppm IBA. Next, we performed transcriptomic analysis of sugarcane buds using RNA sequencing and compared their gene expression during root development due to affect of IBA (100 ppm). A total of 113,475 unigenes were annotated with an average length of 836 bp (N50 = 1,536). The comparative RNA-seq study between the control (CK) and IBA-treated (T) buds showed significant differentially expressed unigenes (494 upregulated and 2086 downregulated). The IBA influenced major biological processes including metabolic process, the cellular process, and single-organism process. For cellular component category, cell, cell part, organelle, membrane, and organelle part were mainly affected. In addition, catalytic activity and binding were primarily affected in the molecular function categories. Furthermore, the expression of genes related to plant hormones and signaling pathways was analyzed by qRT-PCR, which was consistent with the RNA-seq expression profile. This study provides new insights into the IBA response to the bud sprouting in sugarcane based on RNA sequencing, and generated information could help further research on breeding improvement of sugarcane.

11.
Ultrastruct Pathol ; 35(4): 155-61, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21657817

RESUMO

To further understand the pathological characteristics of multiple organ involvement of the 2009 pandemic influenza A/H1N1 infection, tissues of bronchial mucosa, lung, myocardium, gastrocnemius, and liver from 3 patients with fatal A/H1N1 infections were investigated by light microscopy and transmission electron microscopy. In all 3 patients, bronchial mucosa showed necrotizing bronchiolitis, epithelial necrosis and desquamation, and squamous metaplasia, while lung consolidation or fibrosis was identified. Myocardium and gastrocnemius exhibited focal necrosis and fibrosis, surrounded by muscle cells showing features of cell damage. In liver, there was widespread fatty degeneration and necrosis, most often around the central lobular vein and portal area. Viral particles were found in all samples, frequently located in endothelium, epithelium, and muscle cells. The observations demonstrate that in fatal cases of A/H1N1 infection, viruses not only infect the respiratory system, but also engage in multiple organ invasions, causing pathologic changes.


Assuntos
Interações Hospedeiro-Patógeno , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/patologia , Insuficiência de Múltiplos Órgãos/patologia , Pandemias , Adulto , Idoso , Brônquios/patologia , Brônquios/virologia , Bronquiolite/patologia , Bronquiolite/virologia , China/epidemiologia , Fígado Gorduroso/patologia , Fígado Gorduroso/virologia , Fibrose/patologia , Fibrose/virologia , Coração/virologia , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/ultraestrutura , Influenza Humana/mortalidade , Influenza Humana/virologia , Pneumopatias/patologia , Pneumopatias/virologia , Masculino , Microscopia Eletrônica de Transmissão , Insuficiência de Múltiplos Órgãos/mortalidade , Insuficiência de Múltiplos Órgãos/virologia , Músculo Esquelético/patologia , Músculo Esquelético/virologia , Miocárdio/patologia , Necrose/patologia , Necrose/virologia , Mucosa Respiratória/ultraestrutura , Mucosa Respiratória/virologia , Taxa de Sobrevida
13.
Zhonghua Yi Xue Za Zhi ; 90(38): 2708-12, 2010 Oct 19.
Artigo em Zh | MEDLINE | ID: mdl-21162903

RESUMO

OBJECTIVE: To study the effects of drynaria total flavonoid on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) at different glucose concentrations. METHODS: BMSCs of SD rats were isolated, cultivated in vitro, and divided into 6 groups to be induced to differentiate into osteoblasts under different conditions: (1) low glucose control group, (2) high glucose control group, (3) low glucose classical induction group (sodium glycerophosphate+vitamin C+dexamethasone), (4) high glucose classical induction group (sodium glycerophosphate+vitamin C+dexamethasone), (5) low glucose+drynaria total flavonoid group, and (6) high glucose with drynaria total flavonoid group. Alkaline phosphate (ALP) test kit was used to examine the level of ALP. The ALP staining positive rate was examined with modified calcium cobalt method. Alizarin red staining was adopted to observe the number of calcium nodes. Immunohistochemistry was used to detect type I collagen level. Advanced glycosylation end products (AGEs) were tested by ELISA. RESULTS: The A value indicating the ALP activity, ALP staining positive rate, calcium node number, and type I collagen expression score of the low glucose+drynaria total flavonoid group were (0.439±0.024), 48.7%, (9.75±1.71) nodes/HP, and (2.21±0.07) respectively, all significantly higher than those of the sodium glycerophosphate+vitamin C+dexamethasone [(0.385±0.029), 35.0%, (6.25±0.96) nodes/HP, and (1.93±0.13) respectively, all P<0.05]. The A value, ALP staining positive rate, calcium node number, and type I collagen expression score of the high glucose with drynaria total flavonoid group were (0.352±0.022), 25.3%, (4.50±1.29)/HP, and (1.70±0.03) respectively, all significantly higher than those of the sodium glycerophosphate+vitamin C+dexamethasone [(0.139±0.013), 22.7%, (3.25±1.50)/HP, and (1.28±0.27) respectively, all P<0.05]. The AGE expression levels of the high glucose classical induction group and high glucose+drynaria total flavonoid group were both significantly higher than those of the low glucose classical induction group and low glucose+drynaria total flavonoid group (both P<0.05). There were no significant differences in the AGE level among the low glucose control, low glucose classical induction, and low glucose+drynaria total flavonoid groups (all P<0.05); and among the high glucose control, high glucose classical induction, and high glucose+drynaria total flavonoid groups (all P<0.05). However, the AGE levels of the high glucose groups were all significantly higher than those of the corresponding low glucose groups (all P<0.05). Glucose increased the AGE levels dose- and time-dependently. The concentrations of AGEs were significantly negatively correlated with the expression of type I collagen (r=-0.410, P<0.05). CONCLUSIONS: Drynaria total flavonoid promotes the osteogenic differentiation of BMSCs and relieves the inhibitory effect of osteogenic differentiation by glucose at high concentration. Thus drynaria total flavonoid may provide a potential therapy for diabetic osteoporosis.


Assuntos
Células da Medula Óssea/efeitos dos fármacos , Flavonoides/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Polypodiaceae/química , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Glucose/metabolismo , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Ratos , Ratos Sprague-Dawley
14.
Zhonghua Yu Fang Yi Xue Za Zhi ; 44(10): 923-7, 2010 Oct.
Artigo em Zh | MEDLINE | ID: mdl-21176525

RESUMO

OBJECTIVE: This study was to investigate the growth and proliferation characteristics of rat bone mesenchymal stem cells (BMSCs) isolated by the method of whole bone marrow culture and to explore the effect of cell inoculation density and incubation period on cell proliferation, with an aim to provide multipotential seed cells for preventing from degenerative disease. METHODS: Bone mesenchymal stem cells were isolated by the method of whole bone marrow culture and then cultured in vitro. The cell morphologic features were observed by inverted microscope. The cell surface antigens were identified by flow cytometry. The effect of cell inoculation density and culture period on cell growth and proliferation was explored by analyzing the characteristics of a ten-day cell growth curve in 96-well plates. RESULTS: Flow cytometry results showed the detection rates for CD29, CD34 and CD45 were 97.68% (7607/7788), 7.93% (661/8340) and 2.76% (215/7788) respectively, which was consistent with the expression characteristics of BMSCs surface antigens. BMSCs became uniform after three cell passages, existing in a typical shape of whirlpool or radial colony. The senescent cells started to appear at 7(th) passage, and more senescent cells were found at 10(th) passage. The growth curve for moderate inoculation density was typically S-shaped. Lag phase was found during the first two days, and logarithm growth phase was in the following three days. Plateau phase started from the 6(th) day and cell numbers decreased slightly from the 8(th) day. CONCLUSION: The whole bone marrow culture is an effective way to obtain BMSCs. A moderate inoculation density was more advantageous to cell proliferation, by which more seed cells could be obtained.


Assuntos
Células da Medula Óssea/citologia , Proliferação de Células , Células-Tronco Mesenquimais/citologia , Animais , Diferenciação Celular , Células Cultivadas , Citometria de Fluxo , Masculino , Ratos , Ratos Sprague-Dawley
15.
Ultrastruct Pathol ; 33(3): 136-40, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19479654

RESUMO

Primary and metastatic carcinomas have a reactive stroma characterized by many myofibroblasts. These cells have also been documented in nonepithelial malignancies, such as sarcomas, malignant melanoma, and lymphoid tumors but in generally far fewer numbers. In non-Hodgkin's lymphoma, Hodgkin's disease, and leukemia, myofibroblasts are rather rarely documented. In particular, there appear to be no reports of myofibroblasts in either primary bone-marrow/peripheral blood leukemia or secondary deposits of leukemia. In this paper, a case of a relapsed chronic myeloid leukemia appearing in an inguinal lymph node is described, containing many myofibroblasts. The case is detailed and presented with a discussion on the role of myofibroblasts in the progression of nonepithelial cancers.


Assuntos
Transformação Celular Neoplásica/ultraestrutura , Fibroblastos/ultraestrutura , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Linfonodos/ultraestrutura , Miócitos de Músculo Liso/ultraestrutura , Progressão da Doença , Feminino , Humanos , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Organelas/ultraestrutura , Recidiva
16.
Ultrastruct Pathol ; 33(5): 236-42, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19895296

RESUMO

Severe malarial anemia causes considerable mortality and morbidity in endemic areas. Possible mechanisms underlying the anemia include lysis of parasitized and nonparasitized red cells as well as parasite product-mediated effects on erythropoiesis. The latter include suppression of erythropoiesis, dyserythropoiesis, and ineffective erythropoiesis. Present transmission electron microscope data in two cases of Pasmodium vivax malaria show a hitherto undescribed mechanism contributing to malarial anemia, namely, infection of erythroblasts by parasites and their subsequent degradation. No parasites were detected in the peripheral blood but parasites were found in the bone marrow. These findings emphasise the value of bone marrow examination in the diagnosis and eradication of malaria.


Assuntos
Anemia/parasitologia , Eritroblastos/parasitologia , Malária/parasitologia , Plasmodium vivax/fisiologia , Adulto , Anemia/patologia , Células da Medula Óssea/parasitologia , Células da Medula Óssea/patologia , Eritroblastos/patologia , Eritropoese/fisiologia , Feminino , Hemólise , Interações Hospedeiro-Parasita , Humanos , Malária/patologia , Masculino , Plasmodium vivax/isolamento & purificação , Plasmodium vivax/ultraestrutura
17.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 27(2): 311-317, 2019 Apr.
Artigo em Zh | MEDLINE | ID: mdl-30998131

RESUMO

OBJECTIVE: To screen and verify the differentially expressed genes related with aging of bone marrow mesenchymal stem cells (BM-MSCs) in acute myeloid leukemia (AML) patients by bioinformatics, so as to provide new molecular markers for the research and clinical treatment of AML. METHODS: The gene expression profiling chip related with BM-MSCs in AML patients in our hospital and the gene chip GSE84881 selected from NCBI database GEO were used for data analysis and exploration. The DAVID analysis software was used to perform gene ontology (GO) enrichment analysis and KEGG pathway enrichment analysis. Furthermore, the differentially expressed genes related with aging of BM-MSCs in AML patients were identified. Bone marrow samples were collected and MSCs were amplified in vitro, and RT-PCR was used to verify the differentially expressed genes, which should be further identified with senescence-associated ß-galactosidase staining and MTT cell proliferation assays. RESULTS: A total of 247 differentially expressed genes were screened out by bioinformatics methods, including genes of 132 up-regulated expression and 115 down-regulated expression. Six differentially expressed genes related with aging of BM-MSCs in AML patients were screened out, including the genes of up-regulated expression, COL3A1 (P<0.05), CRYAB (P<0.01), DCN (P<0.05), and the genes of down-regulated expression, including CCL2 (P<0.05), CTSC (P<0.01) and IL6 (P<0.05). These 6 differentially expressed genes were consistent with data from chip assays, and which was significantly correlated with aging of BM-MSCs in AML patients. Meanwhile, the positive rate of senescence-associated ß-galactosidase staining in BM-MSCs of AML patients was significantly different from that of healthy donors (P<0.01). MTT cell proliferation assay showed that BM-MSCs in AML patients had proliferative ability lower than the healthy donors' BM-MSCs. CONCLUSION: The data here suggest novel clues for the clinical research and treatment of BM-MSCs aging in AML patients.


Assuntos
Leucemia Mieloide Aguda , Células-Tronco Mesenquimais , Células da Medula Óssea , Proliferação de Células , Células Cultivadas , Biologia Computacional , Humanos
18.
Ultrastruct Pathol ; 31(5): 327-32, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17963181

RESUMO

There are few transmission electron microscopic studies on bone marrow biopsies of patients with hematological disease owing to the difficulty of overcoming the artifacts of decalcification. Following the fixation of bone marrow biopsies thoroughly before a mild decalcification procedure, ultrastructural studies were performed on 13 patients with varied hematological diseases. Notable features included blood cell disorganization, fibroblast activation, myofibroblast transformation, as well as accumulation of collagen and extracellular amorphous matrix. In addition, excessive blood cell death in leukemia, apoptosis, and macrophage phagocytosis in myelodysplastic syndrome and polycythemia vera, as well as degranulation of eosinophils and megakaryocytes in chronic idiopathic myelofibrosis were predominant, respectively. The observations suggest that polyclonal fibroblast proliferation and extracellular matrix accumulation may result from inflammation resulting from excessive cell death and active material release of blood cells in the bone marrow of patients with hematological disease.


Assuntos
Células da Medula Óssea/ultraestrutura , Medula Óssea/ultraestrutura , Doenças Hematológicas/patologia , Microscopia Eletrônica de Transmissão/métodos , Adolescente , Adulto , Biópsia , Criança , Doença Crônica , Estruturas Citoplasmáticas/ultraestrutura , Matriz Extracelular/ultraestrutura , Feminino , Fibroblastos/ultraestrutura , Doenças Hematológicas/genética , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologia , Leucemia Plasmocitária/genética , Leucemia Plasmocitária/patologia , Masculino , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/genética , Síndromes Mielodisplásicas/patologia , Policitemia Vera/genética , Policitemia Vera/patologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Mielofibrose Primária/genética , Mielofibrose Primária/patologia
19.
Chin Med J (Engl) ; 119(18): 1536-42, 2006 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-16996007

RESUMO

BACKGROUND: Sam68 plays an important role as a multiple functional RNA binding nuclear protein in cell cycle progress, RNA usage, signal transduction, and tyrosine phosphorylation by Src during mitosis. However, its precise impact on these essential cellular functions remains unclear. The purpose of this study is to further elucidate Sam68 functions in RNA metabolism, signal transduction regulation of cell growth and cell proliferation in DT40 cell line. METHODS: By using gene targeting method, we isolated a mutation form of Sam68 in DT40 cells and described its effect on cell growth process and signal transduction. Southern, Northern, and Western blot, phosphorylation and flow-cytometric analyses were performed to investigate the Sam68 functions. RESULTS: A slower growth rate (2.1 hours growth elongation) and longer S phase (1.7 hours elongation) was observed in the Sam68 mutant cells. Serum depletion resulted in increased amounts of dead cells, and expansion of S phase in mutant cells. Upon B cell cross-linking, the maximal level of tyrosine phosphorylation on BLNK was observed to be significantly lower in mutant cells. CONCLUSIONS: The proline rich domain of Sam68 is involved in cell growth control by modulating the function of mRNAs in S phase or earlier and the functions as an adaptor molecule in B cell signal transduction pathways.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Linfócitos B/fisiologia , Processos de Crescimento Celular/fisiologia , Proteínas de Ligação a RNA/metabolismo , Transdução de Sinais/fisiologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/fisiologia , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Sítios de Ligação/genética , Western Blotting , Ciclo Celular/fisiologia , Morte Celular/fisiologia , Processos de Crescimento Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Meios de Cultura Livres de Soro/farmacologia , Mutação/genética , Fosforilação , Prolina/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/fisiologia , Receptores de Antígenos de Linfócitos B/imunologia , Receptores de Antígenos de Linfócitos B/fisiologia , Transdução de Sinais/efeitos dos fármacos , Tirosina/metabolismo
20.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 22(3): 671-4, 2014 Jun.
Artigo em Zh | MEDLINE | ID: mdl-24989274

RESUMO

The study was aimed to investigate the effect of CIAPIN1 gene on the proliferation of chronic myeloid leukemia (CML) cell line K562. The shRNA eukaryotic expression vector targeting CIAPIN1 gene was constructed and transfected into K562 cells. The inhibitory efficiency on K562 cells was detected by real-time PCR and Western blot; the proliferative activity of K562 cells was detected by MTT assay; the number and size of colonies were assessed by using colony-forming test; the tumorigenic potential was tested in vivo by using nude mice. The results indicated that as compared with control group, the CIAPIN1 gene expression statistically decreased; the proliferative activity of K562 cells in interference group was distinctly weakened; the number and size of colonies were significantly reduced; the tumorigenic potential was also lowered in vivo. It is concluded that inhibition of CIAPIN1 expression can inhibit K562 cell proliferation in vitro and in vivo.


Assuntos
Proliferação de Células , Peptídeos e Proteínas de Sinalização Intracelular/genética , Vetores Genéticos , Humanos , Células K562 , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , RNA Interferente Pequeno , Transfecção
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