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Objective: To investigate the role of RNA m6A methylation in mediating cerebellar dysplasia through analyzing the phenotypes of the mouse cerebella and the expression of several key m6A regulators upon hypobaric hypoxia treatment. Methods: Five-day old C57/BL6 mice were exposed to hypobaric hypoxia for 9 days. The status of mouse cerebellar development was analyzed by comparing the body weights, brain weights and histological features. Immunostaining of cell-type-specific markers was performed to analyze the cerebellar morphology. Real-time PCR, Western blot and immunohistochemical staining were performed to detect the expression of key m6A regulators in the mouse cerebella. Results: Compared with the control, the body weights, brain weights and cerebellar volumes of hypobaric hypoxic mice were significantly reduced (P<0.01). The expression of specific markers in different cells, including NeuN (mature neuron), Calbindin-D28K (Purkinje cell) and GFAP (astrocyte), was decreased in hypobaric hypoxic mouse cerebella (P<0.01), accompanied with disorganized cellular structure. The expression of methyltransferase METTL3 was significantly down-regulated in the cerebella of hypobaric hypoxic mice (P<0.05). Conclusions: Hypobaric hypoxia stimulation causes mouse cerebellar dysplasia, with structural abnormalities in mature granular neurons, Purkinje cells and astrocytes. Expression of METTL3 is decreased in hypobaric hypoxic mice cerebellum compared with that of normobaric normoxic mice, suggesting that its mediated RNA m6A methylation may play an important role in hypobaric hypoxia-induced mouse cerebellar dysplasia.
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Calbindinas , Cerebelo , Proteínas de Unión al ADN , Hipoxia , Metiltransferasas , Ratones Endogámicos C57BL , Proteínas del Tejido Nervioso , Células de Purkinje , Animales , Ratones , Cerebelo/metabolismo , Hipoxia/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Proteínas del Tejido Nervioso/genética , Células de Purkinje/metabolismo , Células de Purkinje/patología , Calbindinas/metabolismo , Calbindinas/genética , Metiltransferasas/metabolismo , Metiltransferasas/genética , Proteína Ácida Fibrilar de la Glía/metabolismo , Proteína Ácida Fibrilar de la Glía/genética , Astrocitos/metabolismo , Regulación hacia Abajo , Metilación , Adenosina/metabolismo , Adenosina/análogos & derivados , Malformaciones del Sistema Nervioso/metabolismo , Malformaciones del Sistema Nervioso/genéticaRESUMEN
Granular cell tumor (GCT) is a relatively rare tumor that develops in soft tissues at various sites in the body, and GCT originating in the bronchus is rather rare. Here, we reported a case of primary GCT of the bronchial to improve the understanding of this disease.
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Tumor de Células Granulares , Humanos , Tumor de Células Granulares/patología , Bronquios/patologíaRESUMEN
Objective: To investigate the role and mechanism of tumor-derived mesenchymal stem cells in regulating the M2 polarization of macrophages within gastric cancer microenvironment. Methods: Gastric cancer tissues and the adjacent non-cancerous tissues were collected from patients underwent gastric cancer resection in the First People's Hospital of Lianyungang during 2018. In our study, THP-1-differentiated macrophages were co-cultured with gastric cancer-derived mesenchymal stem cells (GC-MSCs). Then, the M2 subtype-related gene, the markers expressed on cell surface and the cytokine profile were analyzed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), flow cytometry and Luminex liquid chip, respectively. The key cytokines mediating the inducing effect of GC-MSCs on macrophage polarization into the M2 subtype were detected and screened by Luminex liquid chip, which were further confirmed by the neutralizing antibody test. The expressions of macrophage proteins involved in M2 polarization-related signaling pathways under the different co-culture conditions of GC-MSCs were detected by western blot. Results: In Mac+ GC-MSC-culture medium (CM) group, the expression levels of Ym-1 and Fizz-1 (1.53±0.32 and 13.22±1.05, respectively), which are markers for M2 subtype, were both significantly higher than those of Mac group (1.00±0.05 and 1.21±0.38, respectively, P<0.05). The level of iNOS in Mac+ GC-MSC-CM group (0.60±0.41) was significantly lower than that of Mac group (1.06±0.38, P=0.023). In Mac+ GC-MSC-Transwell (TW) group, the expression levels of Ym-1 and Fizz-1 (1.47±0.09 and 13.16±2.77, respectively) were both significantly higher than those of Mac group (1.00±0.05 and 1.21±0.38, respectively, P<0.05). The level of iNOS in Mac+ GC-MSC-CM group (0.56±0.03) was significantly lower than that of Mac group (1.06±0.38, P=0.026). The ratios of CD163(+) /CD204(+) cells in Mac+ GC-MSC-CM and Mac+ GC-MSC-TW groups (3.80% and 4.40%, respectively) were both remarkably higher than that of Mac group (0.60%, P<0.05). The expression levels of IL-10, IL-6, MCP-1 and VEGF in Mac+ GC-MSC-CM group were (592.60±87.52), (1 346.80±64.70), (11 256.00±29.03) and (1 463.90±66.67) pg/ml, respectively, which were significantly higher than those of Mac group [(41.03±2.59), (17.35±1.79), (5 213.30±523.71) and (267.12±12.06) pg/ml, respectively, P<0.05]. The levels of TNF-α, IP-10, RANTES and MIP-1α were (95.57±9.34), (410.48±40.68), (6 967.30±1.29) and (1 538.70±283.04) pg/ml, which were significantly lower than those of Mac group [(138.01±24.31, (1 298.60±310.50), (14 631.00±4.21) and (6 633.20±1.47) pg/ml, respectively, P<0.05]. The levels of IL-6 and IL-8 in GC-MSCs [(11 185.02±2.82) and (12 718.03±370.17) pg/ml, respectively] were both strikingly higher than those of MSCs from adjacent non-cancerous gastric cancer tissues [(270.71±59.38) and (106.04±32.84) pg/ml, repectively, P<0.05]. The ratios of CD86(+) cells in Mac+ IL-6-blocked-GC-MSC-CM and Mac+ IL-8-blocked-GC-MSC-CM groups (28.80% and 31.40%, respectively) were both higher than that of Mac+ GC-MSC-CM group (24.70%). Compared to Mac+ GC-MSC-CM group (13.70%), the ratios of CD204(+) cells in Mac+ IL-6-blocked-GC-MSC-CM and Mac+ IL-8-blocked-GC-MSC-CM groups (9.90% and 8.70%, separately) were reduced. The expression levels of p-JAK2 and p-STAT3, which are proteins of macrophage M2 polarization-related signaling pathway, in Mac+ GC-MSC-CM group (0.86±0.01 and 1.08±0.01, respectively) were significantly higher than those of Mac group (0.50±0.01 and 0.82±0.01, respectively, P<0.05). The expression levels of p-JAK2 in Mac+ IL-6-blocked-GC-MSC-CM group (0.47±0.02) were significantly lower those that of Mac+ GC-MSC-CM group (0.86±0.01, P<0.05). The expression levels of p-JAK2 and p-STAT3 in Mac+ IL-8-blocked-GC-MSC-CM group (0.50±0.01 and 0.85±0.01, respectively) were both significantly lower than those of Mac+ GC-MSC-CM group (0.86±0.01 and 1.08±0.01, P<0.05). The expression levels of p-JAK2 and p-STAT3 in Mac+ IL-6/IL-8-blocked-GC-MSC-CM group (0.37±0.01 and 0.65±0.01, respectively) were both significantly lower than those of Mac+ GC-MSC-CM group (0.86±0.01 and 1.08±0.01, P<0.05). Conclusion: GC-MSCs promote the activation of JAK2/STAT3 signaling pathway in macrophages via high secretions of IL-6 and IL-8, which subsequently induce the macrophage polarization into a pro-tumor M2 subtype within gastric cancer microenvironment.
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Células Madre Mesenquimatosas , Neoplasias Gástricas , Humanos , Interleucina-6/genética , Interleucina-8/metabolismo , Interleucina-8/farmacología , Janus Quinasa 2/metabolismo , Macrófagos/metabolismo , Factor de Transcripción STAT3/metabolismo , Transducción de Señal , Neoplasias Gástricas/patología , Microambiente TumoralRESUMEN
Objective: To investigate the value of chromosomes 7 and 8 polysomy in circulating tumor cells (CTCs) for the diagnosis of non-small cell lung cancer, and the correlation of CTCs with clinical pathological characteristics and epidermal growth factor receptor (EGFR) mutations in cancer tissue. Methods: Fifty-seven patients with non-small cell lung cancer and 21 patients with benign lung diseases were enrolled at Beijing Chaoyang Hospital, Capital Medical University, Beijing, China from November 2017 to October 2020. Negative enrichment combined with immunofluorescence in situ hybridization (imFISH) was used to identify CTCs polysomy on chromosomes 7 and 8. EGFR mutations in 56 lung cancer patients was detected using ARMS-PCR. Results: CTCs were detected in 93.0% (53/57) of non-small cell lung cancers and 28.6% (6/21) benign lung lesions. The difference between lung cancer patients and the control cohort was statistically significant (P<0.01). Receive operator curve (ROC) analyses showed that, when the cut-off value was 1 cell/3.2 mL, Youden index had the highest sensitivity of 93.0% and specificity of 71.4% (AUC=0.906, 95%CI:0.833-0.980, P<0.01). The positive rate of CTCs in stage â ¢-â £ cancers was significantly higher than that in stage â -â ¡ (P=0.023). No significant correlation was observed between positive rate of CTCs or chromosome polysomy and age, gender, smoking status, pathologic types and EGFR mutation status. The number of CTCs in EGFR mutated group was higher than that in the non-mutated group (6.5±1.1 vs. 3.7±0.7, P=0.045). The detection rate for CTCs ≥5 in the EGFR mutated group was also higher than the EGFR non-mutated group (52.0% vs. 19.4%,P=0.010). Conclusion: Detection of CTCs with chromosomes 7 and 8 polysomy has potential value in auxiliary diagnosis of non-small cell lung cancer, and the number of CTCs is correlated to TNM stage and EGFR gene mutation status.
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Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Células Neoplásicas Circulantes , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Carcinoma de Pulmón de Células no Pequeñas/genética , China , Receptores ErbB/genética , Humanos , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , MutaciónRESUMEN
Major air pollutants include particulate matter, ozone, sulfur dioxide and nitrogen dioxide, etc. Recent posts have confirmed that air pollution has a variety of adverse health effects on people's health.For professional people, because of occupational hazards of these major atmospheric pollutants also exist in the workplace, is likely to suffer from the double hazards of occupational hazards and air pollutants in the workplace, if similar pollutants are present in the home, the daily exposure concentration of the occupational population may be significantly higher than that of the general population. Exposure limits and testing methods for major atmospheric pollutants (particulate matter or dust, ozone, sulfur dioxide and nitrogen dioxide) are set by relevant standards in workplace air, ambient air and indoor air. However, due to different places and different management departments, there are differences in the detection methods of the same indicators, which brings difficulties to estimate the total daily exposure level. The purpose of this paper is to discuss the "consistency" of the detection method of relevant pollutants in the air, in order to provide scientific basis for estimating the daily exposure level of pollutants in different populations.
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Contaminantes Atmosféricos/análisis , Contaminación del Aire Interior/análisis , Monitoreo del Ambiente/normas , Lugar de Trabajo , Polvo/análisis , Dióxido de Nitrógeno/análisis , Ozono/análisis , Material Particulado/análisis , Dióxido de Azufre/análisisRESUMEN
Objective: To establish a solvent desorption gas chromatography method for determination of cyclohexene in workplace air. Methods: Cyclohexene in the air of workplace was collected with carbon tube and desorbed by carbon disulfide. The target toxicant was separated with the GC column and analyzed with FID detector, identified by retention time, and quantified by peak area. Results: The linear range of cyclohexene in the air of workplace was 0.77~4 050.00 µg/ml, with a correlation coefficient of 0.9999. The limit of detection was 0.23 µg/ml. The lower limit of quantification was 0.77 µg/ml. The minimum detectable concentration was 0.15 mg/m(3) under1.5 L sampling volume and 1.0 ml extraction solution volume. The within-run precision of different cyclohexene concentrations was 0.62%~1.9% and the between-run precisions was 1.5%~3.5%; The average extraction efficiency was 96.4%; Penetration capacity (100 mg of carbon tube) was 29.4 mg; The average collection efficiency was 100%; The samples could be stored for 7 days at room temperature. When placed in 4 â refrigerator, the samples could be stored for 14 days. The potential coexistence of cyclohexane, hexane, benzene, toluene and ethylbenzene with cyclohexene in the air did not interfere with the results of determination. Conclusion: This method has high sensitivity, precision, accuracy and lower limit of detection and it is applicable for determination of cyclohexene in workplace air.
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Contaminantes Ocupacionales del Aire , Ciclohexenos , Lugar de Trabajo , Contaminantes Ocupacionales del Aire/análisis , Cromatografía de Gases , Ciclohexenos/análisis , SolventesRESUMEN
A fluid (a gas or a liquid) adsorbed in a porous material can behave very differently from its bulk counterpart. The advent of various synthesized materials with nanopores and their wide applications have provided strong impetus for studying fluids in confinement because our current understanding is still incomplete. From a large number of Monte Carlo simulations, we found a scaling relation that allows for connecting some thermodynamic properties (chemical potential, free energy per particle, and grand potential per particle) of a confined fluid to the bulk ones. Upon rescaling the adsorbed fluid density, the adsorption isotherms for many different confining environments collapse to the corresponding bulk curve. We also reveal the intimate connection of the reported scaling relation to Gibbs theory of inhomogeneous fluids and morphological thermodynamics. The advance in our understanding of confined fluids, gained from this study, also opens attractive perspectives for circumventing experimental difficulty for directly measuring some fluid thermodynamic properties in nanoporous materials.
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AIM: To identify the basic characteristics and gene expression profiles of supernumerary teeth derived stem cells (SNTSCs) and compare them with those of normal dental pulp stem cells (DPSCs). METHODOLOGY: Flow cytometry was conducted to identify the protein expression of stem cell markers. Cell proliferation, migration and differentiation abilities of both SNTSCs and DPSCs were determined by CCK8, transwell and differentiation assays, respectively. Gene expression profiles were studied by RNA sequencing analyses. After knocking down the expression of certain differential expression genes (DEGs), the function of DEGs was investigated by CCK8 and transwell assays. Statistical differences were determined using a two-tailed t-test and P values below 0.05 were considered significant. RESULTS: Supernumerary teeth derived stem cells were capable of differentiating into adipocyte, chondrocyte and osteoblast lineage cells, and compared to ordinary DPSCs, SNTSCs had a significantly higher cell proliferation rate (P < 0.01) and significantly lower migration rate (P < 0.01). RNA-seq results revealed the differential expression genes (DEGs) between SNTSCs and DPSCs. A principal component analysis (PCA) and cluster analysis revealed that the gene expression patterns of SNTSCs and DPSCs were different from each other. A total of 12 861 genes were differentially expressed at a significant P value (P ≤ 0.01), and 5292 of these increased in SNTSCs and 7569 decreased. Further study on the selected DEGs revealed that FUT11, FAM155A and BRD2 inhibited the cell proliferation rate of SNTSCs, and FUT11 and GLUD1 inhibited the cell migration rate, whilst FAM155A promoted the migration rate. CONCLUSIONS: The biological characteristics and gene expression profile of SNTSCs was revealed. The stem cell properties of SNTSCs were similar to normal DPSCs but they had a high cell proliferation rate and may have greater potential for cell differentiation.
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Diente Supernumerario , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Pulpa Dental , Humanos , Análisis de Secuencia de ARN , Células MadreRESUMEN
Objective: To investigate the expression and potential role of heterogeneous nuclear ribonucleo-protein A2B1 (HNRNPA2B1) in mouse cerebellar development and the significance of HNRNPA2B1 in human medulloblastoma. Methods: The data of HNRNPA2B1 RNA expression in mouse and human cerebella were obtained from databases. Western blot and immunohistochemical staining were performed to detect the protein level of HNRNPA2B1 in mouse cerebella at different ages. The expression level of HNRNPA2B1 in control human cerebellum and medulloblastoma was detected by immunohistochemical staining. m6A-IP-qPCR method was applied to confirm whether HNRNPA2B1 RNA in Daoy cells was modified with m6A.Western blot was used to detect the effect of MG132 treatment on the HNRNPA2B1 protein level in Daoy cells. Results: The level of HNRNPA2B1 protein in postnatal mouse cerebella was higher than that in adult mouse cerebella, with weak HNRNPA2B1 staining in external granular cells while strong staining in mature Purkinje cells and molecular layer. Compared with control normal human cerebella, the RNA expression level of HNRNPA2B1 increased in medulloblastoma, while immunohistochemical staining showed that the mean intensity of HNRNPA2B1 decreased in medulloblastoma. HNRNPA2B1 RNA in medulloblastoma and Daoy cells was modified by m6A. The HNRNPA2B1 protein level in Daoy cells increased upon MG132 treatment. Conclusions: HNRNPA2B1 is dynamically expressed during mouse cerebellar development. Compared with normal human cerebella, HNRNPA2B1 is significantly up-regulated at transcriptional level but obviously down-regulated at translational level in medulloblastoma. These results indicate that HNRNPA2B1 may be involved in cerebellar development process and medulloblastoma tumorigenesis. The m6A methylation in HNRNPA2B1 transcript and protein ubiquitin-proteasome pathway may account for the down-regulation of HNRNPA2B1 at protein level.
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Neoplasias Cerebelosas , Meduloblastoma , Animales , Línea Celular Tumoral , Cerebelo , Regulación hacia Abajo , Ribonucleoproteína Heterogénea-Nuclear Grupo A-B , Humanos , RatonesRESUMEN
Objective: To discuss the difference between pyrophosphoric acid method and infrared spectrophotometry for the determination of silica content in dust. Methods: The content of silica in the laboratory comparison samples organized by CDC Occupational Health Institute in China in 2018, and purchased quality control samples were determined by pyrophosphate method. Meanwhile, the samples were qualitatively and quantitatively analyzed by infrared spectrophotometry, and the results obtained by the two methods were compared. Results: Four samples (062C1ã062C2ãGDOHZKTG012-1ãGDOHZKTG012-2) were detected by pyrophosphate method and infrared spectrophotometry. The results of pyrophosphate method were 55.49%, 5.24%, 4.90% and 54.72%, respectively. The results of infrared spectrophotometry were 0.91%, 1.87%, 1.29% and 1.16% respectively. Conclusion: The content of silica in dust determined by pyrophosphate method is higher than that by infrared spectrophotometry.
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Contaminantes Ocupacionales del Aire/análisis , Difosfatos , Polvo/análisis , Dióxido de Silicio/análisis , Espectrofotometría Infrarroja , ChinaRESUMEN
Objective: To establish a elution solution-liquid chromatography method for determination of p-Phenylene diamine (PPD) in workplace air. Methods: p-Phenylene diamine (PPD) in the air of workplace was collected with glass fiber filters coated with dilute sulfuric acid and extracted with an aqueous EDTA solution. The target toxicant was separated with the C(18) column and analyzed with UV detector, identified by retention time, and quantified by peak area. Results: The linear range of PPD in the air of workplace was 2.00~10.00 µg/ml, with a correlation coefficient of 0.999 96. The limit of detection was 0.07 µg/ml. The lower limit of quantification was 0.23 µg/ml. The minimum detectable concentration was 0.003 1 mg/m(3) under 45.0 L sampling volume and 2.0 ml extraction solution volume. The within-run precision of different PPD concentrations was 0.15%~2.3% and the between-run precisions was 1.4%~2.6%; The extraction efficiencies was 91.4%~95.4%; The average collection efficiencies was 96.6%; The samples could be stored for 7 days isolation of air. The potential coexistence of m-Phenylene diamine and o-Phenylene diamine with p-Phenylene diamine (PPD) in the air did not interfere with the results of determination. Conclusion: This method has high sensitivity, precision, accuracy and lower limit of detection and it is applicable for determination of p-Phenylene diamine (PPD) in workplace air.
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Contaminantes Ocupacionales del Aire/análisis , Fenilendiaminas/análisis , Cromatografía Liquida/métodos , Humanos , Límite de Detección , Lugar de TrabajoRESUMEN
Objective: To study the effect of p-phenylenediamine (PPD) on liver and kidney function in occupational exposed workers. Methods: Workers in a hair dye production enterprise which used p-phenylenediamine as a raw material for production were selected as the main research population. Then we conducted a questionnaire survey on the basic conditions of workers and conducted occupational health checkups on general health status, liver and kidney function. Occupational health examination assessment results were tested in Taizhou Cancer Hospital. All data was built using EpiData 3.1 software, and statistical analysis was performed using software SPSS 20.0. Results: The liver function indicators including direct bilirubin, prealbumin, total protein, and white protein, globulin, aspartate aminotransferase, glutamyl transpeptidase, and total bilirubin in the workers exposed to high concentration of PPD were at high normal values, and these indicators were significantly different from low PPD concentration group (P<0.05) . The serum creatinine and serum uric acid in the renal function index were significantly higher in workers exposed to PPD than in workers exposed to low concentrations and in the control group (P<0.05) . Conclusion: Occupational exposed to PPD may have a hazard to the workers' liver and kidney function. Long-term occupational exposure to PPD may lead to increased cumulative exposure of workers, which may cause potential chronic liver and kidney damage in occupationally exposed populations.
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Enfermedad Hepática Inducida por Sustancias y Drogas , Enfermedades Renales/inducido químicamente , Industria Manufacturera , Enfermedades Profesionales/inducido químicamente , Exposición Profesional/efectos adversos , Fenilendiaminas/toxicidad , Tinturas para el Cabello , Humanos , Riñón/efectos de los fármacos , Riñón/fisiopatología , Hígado/efectos de los fármacos , Hígado/fisiopatología , Exposición Profesional/estadística & datos numéricos , Factores de TiempoRESUMEN
Objective: To study the effects p-phenylenediamine (PPD) on lung function and health-related quality of life of occupational exposed workers. Methods: This study was based on data from a company that produce hair dye containing PPD in China. Workers who exposed to PPD were selected as the study group, and workers un-exposed to PPD were selected as the control group. Questionnaires on health-related quality of life of workers using the 36-item Short Form Health Survey (SF-36) . Occupational health examination assessment results were tested in Taizhou Cancer Hospital. The lung function test includes forced vital capacity (FVC) , forced expiratory volume in one second (FEV(1.0)) , and ratio of FEV(1.0) to FVC (FEV(1.0)/FVC) . Results: The difference in systolic blood pressure between the PPD exposed group and the control group was statistically significant (P<0.05) . FVC, FEV(1.0), and FEV(1.0)/FVC of the lung function indexes in the exposed group were lower than those in the control group (P<0.05) . In the health-related quality of life, body pain (P=0.002) , general health (P=0.029) , vitality (P=0.038) , and mental health (P=0.003) were lower in the exposed group than in the control group. Conclusion: Occupational exposed to PPD may induce hazard to the workers'lung function and may cause detrimental effect on workers' health-related quality of life.
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Pulmón/efectos de los fármacos , Exposición Profesional/efectos adversos , Fenilendiaminas/toxicidad , Calidad de Vida , Estudios de Casos y Controles , China , Volumen Espiratorio Forzado , Humanos , Pulmón/fisiopatología , Capacidad VitalRESUMEN
It appears to be a common sense to measure the crowdedness of a fluid system by the densities of the species constituting it. In the present work, we show that this ceases to be valid for confined fluids under some conditions. A quite thorough investigation is made for a hard sphere (HS) fluid adsorbed in a hard sphere matrix (a quench-annealed system) and its corresponding equilibrium binary mixture. When fluid particles are larger than matrix particles, the quench-annealed system can appear much more crowded than its corresponding equilibrium binary mixture, i.e., having a much higher fluid chemical potential, even when the density of each species is strictly the same in both systems, respectively. We believe that the insight gained from this study should be useful for the design of functionalized porous materials.
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Objective: To investigate the effect of curcumin on the apoptosis and autophagy of human gastric cancer cells with different degree of differentiation. Methods: Gastric cancer cell lines BGC-823 and MKN-28 were treated with curcumin at different concentrations. The effect of curcumin on cell proliferation was measured by MTT assay. Apoptosis was assessed by flow cytometry. Autophagy status was analyzed by acridine orange staining. The expression levels of apoptotic and autophagy-related proteins were detected by Western blot. Results: The cell viability of BGC-823 and MKN-28 was inhibited by curcumin in a time- and dose-dependent manner. At 48 h after treatment, the IC(50) value of BGC-823 (15.18 µmol/L) was close to that of MKN-28 (15.84 µmol/L), and the difference was not statistically significant (P=0.513). Meanwhile, flow cytometry showed that curcumin induced the apoptosis of gastric cancer cells in a dose-dependent manner. Western blot results showed that the expression of pro-apoptotic proteins bax, active-caspase-3 and active-caspase-9 was significantly increased in BGC-823 and MKN-28 cells, whereas that of the anti-apoptotic protein bcl-2 was strikingly reduced. In addition, the formation of acidic vesicular organelles in cytoplasm, conversion of LC3-â to LC3-â ¡ and increased levels of autophagy-related proteins Beclin1, Atg7 and Atg5-Atg12 were observed in curcumin-treated cells. Moreover, activation of PI3K/Akt/mTOR signaling pathway was also significantly suppressed after curcumin treatment. Blocking autophagy by adding the autophagy inhibitor 3-methyladenine (3-MA) significantly promoted the apoptotic cell death induced by curcumin. Conclusions: Curcumin induces apoptosis and protective autophagy in human gastric cancer cells in vitro. Curcumin combined with autophagy inhibitor may provide a more effective strategy for its clinical application.
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Antineoplásicos/farmacología , Apoptosis , Autofagia , Diferenciación Celular/efectos de los fármacos , Curcumina/farmacología , Neoplasias Gástricas/patología , Proteínas Reguladoras de la Apoptosis , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular , Humanos , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Transducción de Señal , Neoplasias Gástricas/metabolismo , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
Rice stripe virus (RSV) protein P3 is a suppressor of RNA silencing in plants. P3 has been shown by biomolecular fluorescence complementation assay to self-interact in planta but the regions responsible for homotypic interaction have not been determined. Here we analyzed the domains for the self-interaction of P3 by using yeast two-hybrid, co-immunoprecipitation and fluorescence experiments. The results showed that P3 was also able to interact with itself in yeast and insect cells. The domain responsible for P3-P3 interaction was mapped to amino acids 15-30 at the N-terminal region of P3. Furthermore, subcellular localization suggested that the homo-oligomerization was the prerequisite for P3 to form larger protein aggregates in the nucleus of insect cell.
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Tenuivirus/metabolismo , Proteínas Virales/química , Proteínas Virales/metabolismo , Secuencia de Aminoácidos , Animales , Dimerización , Datos de Secuencia Molecular , Unión Proteica , Estructura Terciaria de Proteína , Spodoptera/virología , Tenuivirus/química , Tenuivirus/genética , Proteínas Virales/genéticaRESUMEN
We studied the association between aldehyde dehydrogenase 2 (ALDH2) polymorphism and coronary artery disease (CAD) and clarified the mechanisms underlying this association. We searched the ISI, Medline (Ovid), PubMed, CNKI, Wanfang, and Weipu Databases. Statistical analysis was performed using Revman 5.0 and Stata12.0 softwares. A total of 3305 cases and 5016 controls in 12 case-control studies were included in this meta-analysis. Variant A allele carriers showed a 48% increased risk of CAD compared with homozygote A allele [odds ratio (OR) = 1.48, 95% confidence interval (CI) = 1.18-1.87 for AA + AG vs GG]. In subgroup analysis by gender, significantly elevated risks were found in the mixed group (OR = 1.78, 95%CI = 1.42-2.22) but not in males (OR = 1.12, 95%CI = 0.79-1.57). In subgroup analysis by disease type, significant elevated risks were associated with A allele carriers in myocardial infarction [OR = 1.69, 95%CI = (1.05-2.71)], in coronary heart disease (OR = 1.36, 95%CI = 1.00-1.86), but not in coronary heart disease plus diabetes mellitus subjects (OR = 1.57, 95%CI = 0.58-4.29). Moreover, those with the GG genotype consumed significantly more alcohol than those with the AA/AG genotypes (standard mean deviation: 6.32 g, 95%CI = 2.09-10.55, P = 0.000). ALDH2 polymorphisms may be risk factors for CAD. Moreover, CAD patients with ALDH2 genotypes AG and AA consumed significantly less alcohol than those with GG. To further evaluate gene-gene and gene-environment interactions between ALDH2 polymorphisms and the risk of CAD, more studies with larger groups of patients are required.
Asunto(s)
Aldehído Deshidrogenasa Mitocondrial/genética , Enfermedad de la Arteria Coronaria/genética , Predisposición Genética a la Enfermedad , Polimorfismo Genético , Alelos , Femenino , Estudios de Asociación Genética , Genotipo , Humanos , Masculino , Oportunidad Relativa , Factores de RiesgoRESUMEN
Hemangioblastoma of the central nervous system occurs as sporadic tumors or as a part of von Hippel-Lindau (VHL) disease, an autosomal dominant hereditary tumor syndrome caused by a germline mutation in the VHL tumor suppressor gene. We screened a Chinese family with VHL for mutations in the VHL gene and evaluated a genetic test for diagnosing VHL disease and clinical screening of family members. DNA extracted from the peripheral blood of all live members and from tissue of deceased family members with VHL disease was amplified by polymerase chain reaction to 3 VHL gene exons. Mutations in the amplification products were compared against the Human Gene Mutation Database. The involvement of multiple organs among the kindred with VHL disease was confirmed by medical history and radiography. Of the 12 members of the 4-generation family, 5 were diagnosed with VHL disease. Patient age at the initial diagnosis was 26-36 years (mean = 31 years). The mean time was 15 (11-19 months) from symptom appearance to the first patient visit to the hospital. Sequence analysis revealed that the frameshift mutation 327del C (p.Gly39Alafs*26) in exon 1 affected all family members, but not the healthy individuals or 16 unrelated controls. Members without gene mutation showed no clinical manifestation of VHL disease. We detected a conserved novel frameshift mutation in the VHL gene of the family members that contributes to VHL. DNA analysis of VHL is advantageous for VHL diagnosis. We developed a quick and reliable method for VHL diagnosis.