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1.
Artículo en Inglés | MEDLINE | ID: mdl-38401103

RESUMEN

Objective: To investigate the relationship between apolipoprotein E (ApoE) gene polymorphism and cognitive impairment (PSCI) in patients after acute ischemic stroke (AIS). Methods: A total of 150 AIS patients were treated in Chengde Central Hospital from December 2022 to December 2023 and were selected and divided into a disorder group (n=88) and a normal group (n=62) according to the presence or absence of PSCI. Clinical data of patients in the two groups were collected, ApoE genotype and allele distribution of patients in the disabled group and the normal group were detected, Montreal Cognitive Assessment and Mini-Mental State Examination scores of patients with different ApoE gene subtypes were compared, and the risk factors of PSCI after AIS were analyzed by unconditional Logistic regression. Results: The proportion of patients with acute lesions (≥3.0 cm) and the degree of carotid artery stenosis (moderate, severe, complete occlusion) in the disorder group was higher than that in the normal group, and the National Institutes of Health Stroke Scale score was higher than that in the control group, with statistical significance (P < .05). There were significant differences in the genotype and allelic distribution of ApoE between the two groups (P < .05). In both groups, the highest genotype frequency of ApoE was the ε3/3 homozygous type, which was 47.73% (in the disorder group) and 72.58% (in the normal group) respectively. In contrast, there were no significant differences in the genotype frequencies of ε2/2, ε2/3, ε2/4 and ε4/4 alleles in the two groups (P > .05). This means that in both groups of patients, the frequency of the ApoE ε3/3 genotype was the highest, while the genotype frequencies of ε2/2, ε2/3, ε2/4 and ε4/4 alleles were not significant between the two groups. difference. The distribution differences of these genotypes and alleles may be related to aspects such as disease risk and physiological function, providing valuable information for in-depth exploration of the role of ApoE in patients. The genotype frequency of ε3/3 in the disorder group was lower than that in the normal group. The frequency of the ε3/4 genotype was higher than that of the normal group, and the difference was statistically significant (P < .05). In both groups, the highest allele frequency was ε3 (68.75% in the disorder group and 83.06% in the normal group), and there was no difference in the frequency of ε2 allele between the two groups (P > .05). The frequency of the ε3 allele in the disorder group was lower than that in the normal group, and the frequency of the ε4 allele was higher than that in the normal group, the difference was statistically significant (P < .05). In the patients with cognitive impairment after AIS (disorder group), the MOCA and MMSE scores of patients with different ApoE subtypes (ε2, ε3, ε4) were compared, and the differences among the three groups were statistically significant (P < .05). The MOCA and MMSE scores in the ε4 group were lower than those in the ε2 and ε3 groups. The difference was statistically significant (P < .05). Logistic regression analysis showed that the degree of carotid artery stenosis, NIHSS score, and ApoEε4 gene were independent risk factors for PSCI in patients with AIS (P < .05). Conclusion: APOE gene polymorphism is associated with cognitive impairment in post-AIS patients, and carrying the ApoE Epsilon 4 gene may be associated with PSCI in post-AIS patients.

2.
Small ; 19(3): e2205706, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36408820

RESUMEN

The restricted charge transfer and slow oxygen evolution reaction (OER) dynamics tremendously hamper the realistic implementation of SnS2 photoanodes for photoelectrochemical (PEC) water splitting. Here, a novel strategy is developed to construct interfacial NCuS bonds between NC skeletons and SnS2 (CuNC@SnS2 ) for efficient PEC water splitting. Compared with SnS2 , the PEC activity of CuNC@SnS2 photoelectrode is tremendously heightened, obtaining a current density of 3.40 mA cm2 at 1.23 VRHE with a negatively shifted onset potential of 0.04 VRHE , which is 6.54 times higher than that of SnS2 . The detailed experimental characterizations and theoretical calculation demonstrate that the interfacial NCuS bonds enhance the OER kinetic, reduce the surface overpotential, facilitate the separation of photon-generated carriers, and provide a fast transmission channel for electrons. This work presents a new approach for modulating charge transfer by interfacial bond design in heterojunction photoelectrodes toward promoting PEC performance and solar energy application.

3.
Chemistry ; 29(35): e202300616, 2023 Jun 22.
Artículo en Inglés | MEDLINE | ID: mdl-36994493

RESUMEN

The industrial ammonia synthesis process consumes a lot of energy and causes serious environmental pollution. As a sustainable approach for ammonia synthesis, photocatalytic nitrogen reduction employing water as the reducing agent has a lot of potential. A simple surfactant-assisted solvothermal method is used to synthesize g-C3 N4 nanotubes with flower-like spherical BiOBr grown inside and outside (BiOBr/g-C3 N4 , BC). The hollow tubular structure realizes the full use of visible light by the multi-scattering effect of light. Large surface areas and more active sites for N2 adsorption and activation are present in the distinctive spatially dispersed hierarchical structures. Particularly, the quick separation and transfer of electrons and holes are facilitated by the sandwich tubular heterojunctions and tight contact interface of BiOBr and g-C3 N4 . The maximal NH3 generation rate of the BiOBr/g-C3 N4 composite catalysts can reach 255.04 µmol⋅ g-1 ⋅ h-1 , and it is 13.9 and 5.8 times that of pure BiOBr and g-C3 N4 . This work provides a novel method for designing and constructing unique heterojunctions for efficient photocatalytic nitrogen fixation.


Asunto(s)
Amoníaco , Fijación del Nitrógeno , Adsorción , Electrones
4.
Mater Today (Kidlington) ; 31: 86-99, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32831620

RESUMEN

Magnetic iron oxide nanoparticles (MIONs) are among the first generation of nanomaterials that have advanced to clinic use. A broad range of biomedical techniques has been developed by combining the versatile nanomagnetism of MIONs with various forms of applied magnetic fields. MIONs can generate imaging contrast and provide mechanical/thermal energy in vivo in response to an external magnetic field, a special feature that distinguishes MIONs from other nanomaterials. These properties offer unique opportunities for nanomaterials engineering in biomedical research and clinical interventions. The past few decades have witnessed the evolution of the applications of MIONs from conventional drug delivery and hyperthermia to the regulation of molecular and cellular processes in the body. Here we review the most recent development in this field, including clinical studies of MIONs and the emerging techniques that may contribute to future innovation in medicine.

5.
Wei Sheng Yan Jiu ; 46(6): 981-985, 2017 Nov.
Artículo en Zh | MEDLINE | ID: mdl-29903211

RESUMEN

OBJECTIVE: Direct determination of 35 trace volatile organic compounds in air by portable gas chromatography mass spectrometry( GC-MS). METHODS: The mixed standard gas of different concentration levels was prepared with high purity nitrogen as diluent gas. The samples were injected into the GC-MS by a hand-held probe. Retention time and characteristic ion were used for qualitative analysis, and the internal standard method was usd for quantitation. RESULTS: Under the established experimental conditions, the 35 poisonous substances were separated and determined well. The relative standard deviation was 3. 2%-15. 1%. The average recovery was 75. 1%-121. 4%( n = 6). CONCLUSION: The portable GC-MS method can be used for qualitative and quantitative detection of volatile organic compounds in ambient air.


Asunto(s)
Cromatografía de Gases y Espectrometría de Masas/métodos , Compuestos Orgánicos Volátiles/análisis , Humanos
6.
Wei Sheng Yan Jiu ; 45(2): 268-72, 2016 Mar.
Artículo en Zh | MEDLINE | ID: mdl-27301228

RESUMEN

OBJECTIVE: To establish a portable gas chromatography (GC) method for the on-site rapid determination of epoxyethane, furan, dichloromethane, benzene, toluene in workplace. METHODS: By using dynamic dilutor, the volatile organic compounds (VOCs) standard gas was prepared and drawn to the capillary column of GC (PID) for separation and analysis. Based on the retention time and peak area, the VOCs was identified and quantified. RESULTS: At the conditions of 60 degrees C column temperature and 9psi pre-pressure, The five VOCs were well separated. Good linear ranges were obtained within the ranges of 0.45 - 90.72 mg/m3 for epoxyethane, 0.22 - 44.50 mg/m3 for furan, 2.19 - 437.4 mg/m3 for dichloromethane, 0.32 - 13.29 mg/m3 for benzene, 0.38 - 15.68 mg/m3 for toluene, respectively. The correlation coefficient was not less than 0.999. The detection limits were 0.03, 0.04, 0.2, 0.008 and 0.03 mg/m3, respectively. The RSD were 0.79% - 2.4%, 1.5% - 1.9%, 0.97% - 1.8%, 2.3% - 3.2% and 3.4% - 4.6%, respectively. The average recovery were 101.4% - 108.4% 85.49% - 94.98%, 97.78% - 106.2%, 99.29% - 103.5% and 95.54% - 101.7% respectively. By using the 7890A GC method, the relative tolerance for the same gas samples were 0.091% - 8.8% for epoxyethane, 3.8% - 6.7% for furan, 0.77% - 9.4% for dichloromethane, 0.24% - 1.6% for benzene, 0.31% - 8.0% for toluene. CONCLUSION: The method is portable, accurate, sensitive, rapid, and exhibits good separation and anti-interference ability. This method is suitable for the rapid detection of VOCs in workplace and also provides reference VOC detection method for the occupational health standard.


Asunto(s)
Contaminantes Ocupacionales del Aire/análisis , Compuestos Orgánicos Volátiles/análisis , Lugar de Trabajo , Benceno/análisis , Cromatografía de Gases , Furanos/análisis , Cloruro de Metileno/análisis , Tolueno/análisis
7.
Nucleic Acids Res ; 41(19): e180, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23945944

RESUMEN

Safety and reliability of transgene integration in human genome continue to pose challenges for stem cell-based gene therapy. Here, we report a baculovirus-transcription activator-like effector nuclease system for AAVS1 locus-directed homologous recombination in human induced pluripotent stem cells (iPSCs). This viral system, when optimized in human U87 cells, provided a targeted integration efficiency of 95.21% in incorporating a Neo-eGFP cassette and was able to mediate integration of DNA insert up to 13.5 kb. In iPSCs, targeted integration with persistent transgene expression was achieved without compromising genomic stability. The modified iPSCs continued to express stem cell pluripotency markers and maintained the ability to differentiate into three germ lineages in derived embryoid bodies. Using a baculovirus-Cre/LoxP system in the iPSCs, the Neo-eGFP cassette at the AAVS1 locus could be replaced by a Hygro-mCherry cassette, demonstrating the feasibility of cassette exchange. Moreover, as assessed by measuring γ-H2AX expression levels, genome toxicity associated with chromosomal double-strand breaks was not detectable after transduction with moderate doses of baculoviral vectors expressing transcription activator-like effector nucleases. Given high targeted integration efficiency, flexibility in transgene exchange and low genome toxicity, our baculoviral transduction-based approach offers great potential and attractive option for precise genetic manipulation in human pluripotent stem cells.


Asunto(s)
Baculoviridae/genética , Proteínas de Unión al ADN/metabolismo , Desoxirribonucleasas de Localización Especificada Tipo II/metabolismo , Marcación de Gen/métodos , Células Madre Pluripotentes Inducidas/metabolismo , Transducción Genética , Transgenes , Línea Celular Tumoral , Reparación del ADN por Unión de Extremidades , Proteínas de Unión al ADN/genética , Desoxirribonucleasas de Localización Especificada Tipo II/genética , Vectores Genéticos , Células HEK293 , Humanos , Integrasas/metabolismo , Mutación , Proteínas Recombinantes de Fusión/metabolismo
8.
ACS Omega ; 9(26): 28866-28878, 2024 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-38973832

RESUMEN

To enable rapid and accurate point-of-care DNA detection, we have developed a single-step, amplification-free nucleic acid detection platform, a DNA substrate-mediated autocatalysis of CRISPR/Cas12a (DSAC). DSAC makes use of the trans-cleavage activity of Cas12a and target template-activated DNA substrate for dual signal amplifications. DSAC employs two distinct DNA substrate types: one that enhances signal amplification and the other that negatively modulates fluorescent signals. The positive inducer utilizes nicked- or loop-based DNA substrates to activate CRISPR/Cas12a, initiating trans-cleavage activity in a positive feedback loop, ultimately amplifying the fluorescent signals. The negative modulator, which involves competitor-based DNA substrates, competes with the probes for trans-cleaving, resulting in a signal decline in the presence of target DNA. These DNA substrate-based DSAC systems were adapted to fluorescence-based and paper-based lateral flow strip detection platforms. Our DSAC system accurately detected African swine fever virus (ASFV) in swine's blood samples at femtomolar sensitivity within 20 min. In contrast to the existing amplification-free CRISPR/Dx platforms, DSAC offers a cost-effective and straightforward detection method, requiring only the addition of a rationally designed DNA oligonucleotide. Notably, a common ASFV sequence-encoded DNA substrate can be directly applied to detect human nucleic acids through a dual crRNA targeting system. Consequently, our single-step DSAC system presents an alternative point-of-care diagnostic tool for the sensitive, accurate, and timely diagnosis of viral infections with potential applicability to human disease detection.

9.
J Gene Med ; 15(10): 384-95, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24105820

RESUMEN

BACKGROUND: The AAVS1 locus is viewed as a 'safe harbor' for transgene insertion into human genome. In the present study, we report a new method for AAVS1 targeting in human-induced pluripotent stem cells (hiPSCs). METHODS: We have developed two baculoviral transduction systems: one to deliver zinc finger nuclease (ZFN) and a DNA donor template for site-specific gene insertion and another to mediate Cre recombinase-mediated cassette exchange system to replace the inserted transgene with a new transgene. RESULTS: Our ZFN system provided the targeted integration efficiency of a Neo-EGFP cassette of 93.8% in G418-selected, stable hiPSC colonies. Southern blotting analysis of 20 AASV1 targeted colonies revealed no random integration events. Among 24 colonies examined for mono- or biallelic AASV1 targeting, 25% of them were biallelically modified. The selected hiPSCs displayed persistent enhanced green fluorescent protein expression and continued the expression of stem cell pluripotency markers. The hiPSCs maintained the ability to differentiate into three germ lineages in derived embryoid bodies and transgene expression was retained in the differentiated cells. After pre-including the loxP-docking sites into the Neo-EGFP cassette, we demonstrated that a baculovirus-Cre/loxP system could be used to facilitate the replacement of the Neo-EGFP cassette with another transgene cassette at the AAVS1 locus. CONCLUSIONS: Given high targeting efficiency, stability in expression of inserted transgene and flexibility in transgene exchange, the approach reported in the present study holds potential for generating genetically-modified human pluripotent stem cells suitable for developmental biology research, drug development, regenerative medicine and gene therapy.


Asunto(s)
Baculoviridae/genética , Endonucleasas/genética , Expresión Génica , Vectores Genéticos/genética , Células Madre Pluripotentes Inducidas/metabolismo , Transgenes , Dedos de Zinc/genética , Endonucleasas/metabolismo , Orden Génico , Genes Reporteros , Sitios Genéticos , Recombinación Homóloga , Humanos , Mutagénesis Insercional
10.
RSC Adv ; 13(31): 21550-21557, 2023 Jul 12.
Artículo en Inglés | MEDLINE | ID: mdl-37469963

RESUMEN

A novel cyclic ion chromatography (IC) system was developed for the simultaneous determination of trace disinfection by-products (DBPs) in drinking water. Five DBPs (chlorite, bromate, chlorate, dichloroacetic acid, and trichloroacetic acid) were sensitively determined by large-volume direct injection, and the interferences of dominant inorganic anions present in water were eliminated online through the cyclic determination of the target analytes. Under optimized conditions, the obtained limits of detection (LODs) were in the range of 0.18-1.91 µg L-1 based on a signal-to-noise ratio (S/N) of 3 and an injection volume of 1.0 mL. The RSDs for peak area and retention time were in the range of 0.13-1.03% and 1.24-4.29%, respectively. Satisfactory recoveries between 92.3% and 106.4% were obtained by adding three concentration gradients of standards to the drinking water samples. The proposed method has advantages such as high sensitivity, facile automation, and no sample pretreatment, and might be a promising approach for routine analysis.

11.
Sci Total Environ ; 892: 164511, 2023 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-37257601

RESUMEN

Monoaromatic hydrocarbons (MACHs) are a ubiquitous category of volatile compounds found in various environmental media. Despite their prevalence, systematic studies of MACHs on a large regional scale are still lacking. Herein, a comprehensive investigation of the occurrence, seasonal variations, distribution characteristics, and health risks of MACHs was carried out by analyzing soil samples (372 surface soils and 96 soil columns) from 33 typical industrial parks in the Yangtze River Delta (YRD) region. MACHs were detected in all surface soil samples. BTEXS (benzene, toluene, ethylbenzene, xylene, and styrene) were the five predominant congeners with the highest detection frequencies (90.9 %-100 %), collectively accounting for >78.2 % of the total MACHs content. Higher residual levels of MACHs were observed in winter compared to summer (P < 0.01), with total concentrations of 24 MACHs ranging from 30.9 ng/g to 1536 ng/g (median: 135 ng/g) in winter and 16.3 ng/g to 931 ng/g (median: 87.9 ng/g) in summer. Soils collected from the northeast of Jiangsu Province and the southwest of Anhui Province exhibited relatively higher levels of MACHs. On the basis of principal component analysis, we proposed that industrial emissions and vehicle exhaust may be the main sources of MACHs contamination in the soils of YRD industrial parks. Vertically, the concentrations of total MACHs decreased with the soil depth. Soil organic matter (OM) content and the concentration of MACHs in the surface soil layer (0-15 cm) were significant factors influencing the vertical migration and distribution of MACHs (P < 0.05). It was verified that residual MACHs in the soils posed lower lifetime non-carcinogenic and carcinogenic risks to the inhabitants of the study area. The field study provides valuable evidence for the formulation of MACHs pollution control policies in the YRD region.


Asunto(s)
Contaminantes del Suelo , Suelo , Monitoreo del Ambiente , Ríos , Estaciones del Año , Contaminantes del Suelo/análisis , China , Medición de Riesgo , Hidrocarburos/análisis , Tolueno/análisis
12.
Artículo en Zh | MEDLINE | ID: mdl-22804996

RESUMEN

OBJECTIVE: To establish a method to detect N-methylacetamide (NMAC) concentration in urine of workers occupationally exposed to NMAC with directly injecting the sample into capillary gas chromatography. METHODS: After frozen urine samples were isolated from precipitation by centrifugation, the aliquot of supernatant was pretreated by protein precipitation with dilution of methanol. The methanol supernatant was separated by Polyethylene Glycol (PEG) capillary columns and detected by nitrogen phosphorous detector (NPD). RESULTS: Good linearity was obtained in the concentration range of 1.0 ∼ 250 mg/L. The correlation coefficient was 1.0000. The minimum detection limit of NMAC in urine was 0.2 mg/L. The method recovery rates were 96.0% ∼ 99.4% at three different concentrations. The mean recovery rate was 97.8%. The relative standard deviations (RSD) of intra- and inter-day were between 1.5% ∼ 3.4%. CONCLUSION: The method was simple, rapid, selective and sensitive and was applicable to detect the urinary NMAC concentration for monitoring occupational exposure levels.


Asunto(s)
Acetamidas/orina , Cromatografía de Gases/métodos , Exposición Profesional/análisis , Monitoreo del Ambiente/métodos , Humanos
13.
Artículo en Zh | MEDLINE | ID: mdl-23257108

RESUMEN

OBJECTIVE: To establish Biological Limit Value (BLV) for N, N-dimethylacetamide (DMAC). METHOD: 201 workers in 3 spandex factories exposed to DMAC were recruited. Air samples were collected using personal air samplers, and urine samples from each works were collected at the end of shift at end of workweek. The urinary metabolite NMAC and air samples of DMAC were determined by gas chromatography (GC). Percentile and relative internal exposure (RIE) were analyzed and proposed a BLV for DMAC. RESULTS: The number of workers who exposure to DMAC below OELs were 133 (66.2%) among 201 workers monitored. Geometric mean (range) concentration of DMAC in air was 19.4 (0.40 ∼ 300.12) mg/m(3), and that of NMAC in urine was 23.7 (1.30 ∼ 189.42) mg/g Cr. A linear correlation was found between the personal air DMAC and creatinine-adjusted NMAC levels in urine collected at the end of shift at end of workweek (F = 188.872, R(2) = 0.487,P < 0.001). The relationship can be described by the equation Log (NMAC mg/g Cr) = 0.685 + 0.455 log (DMAC mg/m(3)). According to the equation the current China OELs value of 20 mg/m(3) would lead to a mean NMAC concentration of 18.92 mg/g Cr. The 90th percentile biomonitoring result below 20 mg/m(3) 8-hour TWA is 23.9 mg MMAC mg/g Cr, and that of NMAC in urine calculated by relative internal exposure (RIE) was 19.0 mg/g Cr. CONCLUSION: A BLV of 20 mg/g Cr NMAC in urine at the end of shift at end of workweek for DMAC was recommend by reference to official values from other countries.


Asunto(s)
Acetamidas/orina , Contaminantes Ocupacionales del Aire/análisis , Exposición Profesional/análisis , Acetamidas/análisis , Adulto , Cromatografía de Gases , Femenino , Humanos , Masculino , Valores Limites del Umbral
14.
Mol Ther Nucleic Acids ; 27: 1078-1091, 2022 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-35228901

RESUMEN

Genetic lineage tracing is indispensable to unraveling the origin, fate, and plasticity of cells. However, the intrinsic leakiness in the CreER-loxP system raises concerns on data interpretation. Here, we reported the generation of a novel dual inducible CreER-loxP system with superior labeling characteristics. This two-component system consists of membrane localized CreER (mCreER: CD8α-FRB-CS-CreER) and TEV protease (mTEVp: CD8α-FKBP-TEVp), which are fusion proteins incorporated with the chemically induced dimerization machinery. Rapamycin and tamoxifen induce sequential dimerization of FKBP and FRB, cleavage of CreER from the membrane, and translocation into the nucleus. The labeling leakiness in Ad293 cells reduced dramatically from more than 70% to less than 5%. This tight labeling feature depends largely on the association of mCreER with HSP90, which conceals the TEV protease cutting site between FRB and CreER and thus preventing uninduced cleavage of the membrane-tethering CreER. Membrane-bound CreER also diminished significantly cytotoxicity. Our studies showed mCreER under the control of the rat insulin promoter increased labeling specificity in MIN6 islet beta-cells. Viability and insulin secretion of MIN6 cells remained intact. Our results demonstrate that this novel system can provide more stringent temporal and spatial control of gene expression and will be useful in cell fate probing.

15.
Wei Sheng Yan Jiu ; 39(6): 770-3, 2010 Nov.
Artículo en Zh | MEDLINE | ID: mdl-21351652

RESUMEN

OBJECTIVE: To establish solvent desorption gas chromatographic method for determination of hydrogenated terphenyl in workplace air. METHODS: According to standards of methods for determination of toxic substances in workplace air, hydrogenated terphenyl in the air was first adsorbed by activated carbon, desorbed by carbon disulfide, and determined by gas chromatography, then quality was used with retention time and quantitation was used by the sums of peaks area. RESULTS: The method presented linear relationship within the ranges of 6-60 microg/ml. The relative standard deviations (RSD) were 1.3%-2.0% and the detection limits were 1.2 microg/ml. The desorption efficiencies were 89.3%-95.4%. Sampling efficiencies were 100% and the breakthrough volumes were beyond 6.7 mg in a 100 mg active carbon with glass wool. The samples in active carbon tubes could be stored for 7 days at room temperature and its repetition was good. Other solvents in the air such as coexisting diphenyl, ortho-, meta-, para-terphenyls had no interferences in this method. CONCLUSION: The method could meet with the requirements of Standards of methods for determination of toxic substances in workplace air and be feasible for determination of hydrogenated terphenyl in workplace air.


Asunto(s)
Contaminantes Ocupacionales del Aire/análisis , Monitoreo del Ambiente/métodos , Compuestos de Terfenilo/análisis , Lugar de Trabajo , Cromatografía de Gases/métodos
16.
Int J Artif Organs ; 32(4): 224-31, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19569030

RESUMEN

The objective of this study was to characterize the levels of free radicals in serum and antioxidase activity after microcapsules were implanted into the subcutaneous space of mice. cell viability was evaluated using ao/Eb staining. serum free radicals, malondialdehyde and superoxide dismutase levels were evaluated by colorimetry analysis. the mice were divided into three groups: saline injection group (n=15), empty microcapsules injected group (n=21), encapsulated cells injected group (n=21). cell viability and serum analysis were executed at 1, 4 and 7 days post-implantation. Hydrogen peroxide and malondialdehyde levels initially increased in the recipients of the empty microcapsules, before decreasing to the basal level. However, in mice receiving the encapsulated cells, the levels were higher at the end of study. nitric oxide and superoxide dismutase increased after the implantation of microcapsules with or without the bHK-21 cells, but were not changed in response to the saline injection. the viability of the encapsulated cells was high in vivo, although some microcapsules had broken by 7 days post-implantation. these results suggest that nitric oxide plays a role in the specific response to microcapsules. the levels of free radicals rapidly increased immediately following microcapsule transplantation, but they caused only slight cellular damage before the microencapsulated cells were exposed.


Asunto(s)
Alginatos/administración & dosificación , Quitosano/administración & dosificación , Radicales Libres/sangre , Naranja de Acridina , Animales , Cápsulas/administración & dosificación , Células Cultivadas , Etidio , Colorantes Fluorescentes , Hipodermoclisis , Malondialdehído/sangre , Ratones , Óxido Nítrico/sangre , Superóxido Dismutasa/sangre
17.
Nat Biomed Eng ; 3(2): 126-136, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30944431

RESUMEN

The potential of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 9 (Cas9)-based therapeutic genome editing is hampered by difficulties in the control of the in vivo activity of CRISPR-Cas9. To minimize any genotoxicity, precise activation of CRISPR-Cas9 in the target tissue is desirable. Here, we show that, by complexing magnetic nanoparticles with recombinant baculoviral vectors (MNP-BVs), CRISPR-Cas9-mediated genome editing can be activated locally in vivo via a magnetic field. The baculoviral vector was chosen for in vivo gene delivery because of its large loading capacity and ability to locally overcome systemic inactivation by the complement system. We demonstrate that a locally applied magnetic field can enhance the cellular entry of MNP-BVs, thereby avoiding baculoviral vector inactivation and causing a transient transgene expression in the target tissue. Because baculoviral vectors are inactivated elsewhere, gene delivery and in vivo genome editing via MNP-BVs are tissue specific.


Asunto(s)
Proteína 9 Asociada a CRISPR/metabolismo , Sistemas CRISPR-Cas/genética , Edición Génica , Fenómenos Magnéticos , Nanopartículas de Magnetita/química , Animales , Baculoviridae/genética , Línea Celular Tumoral , Endocitosis , Femenino , Vectores Genéticos/metabolismo , Humanos , Hígado/metabolismo , Luciferasas/metabolismo , Campos Magnéticos , Nanopartículas de Magnetita/ultraestructura , Ratones Desnudos , Transgenes
19.
Methods Mol Biol ; 1498: 3-21, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-27709565

RESUMEN

The CRISPR/Cas9 system is a powerful tool for precision genome editing. The ability to accurately modify genomic DNA in situ with single nucleotide precision opens up new possibilities for not only basic research but also biotechnology applications and clinical translation. In this chapter, we outline the procedures for design, screening, and validation of CRISPR/Cas9 systems for targeted modification of coding sequences in the human genome and how to perform genome editing in induced pluripotent stem cells with high efficiency and specificity.


Asunto(s)
Sistemas CRISPR-Cas/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , Genoma Humano/genética , Células Madre Pluripotentes/fisiología , Edición de ARN/genética , Línea Celular , Edición Génica/métodos , Células HEK293 , Humanos
20.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 27(3): 321-4, 2005 Jun.
Artículo en Zh | MEDLINE | ID: mdl-16038268

RESUMEN

OBJECTIVE: To determine the physiological role of D-bifunctional protein (DBP) in bile acid biosynthesis through investigating the effect of increasing activity of DBP on bile acid biosynthesis. METHODS: Twenty male Wistar rats were divided into two groups: diethylhexyl phthalate (DEHP) group (n = 10) and control group (n = 10). Serum triglyceride, total cholesterol, hepatic DBP activity, and fecal bile acids were assayed. The mRNA levels of hepatic peroxisome proliferator-activated receptor alpha (PPARalpha), DBP, and cholesterol 7alpha-hydroxylase (CYP7A1) were detected by RT-PCR. RESULTS: Compared with control group, serum triglyceride level was decreased significantly and PPARalphamRNA level was increased significantly in DEHP group (P < 0.01). Together with a sharp induction of DBP mRNA expression and DBP activity in DEHP group (P < 0.01), the levels of CYP7A1 mRNA and fecal bile acids were significantly increased by 1.9 times and 1.6 times respectively compared to control group (P < 0.01). There was a significantly positive correlation between DBP mRNA level or DBP activity and CYP7A1 mRNA level (r = 0.89, P < 0.01; r = 0.95, P < 0.01). CONCLUSION: The up-regulation of DBP mRNA and activity in liver can result in the increase in CYP7A1 mRNA expression and bile acid biosynthesis, suggesting that DBP may be involved in bile acid biosynthesis together with CYP7A1.


Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/metabolismo , Ácidos y Sales Biliares/biosíntesis , Enoil-CoA Hidratasa/metabolismo , Hígado/metabolismo , Complejos Multienzimáticos/metabolismo , Animales , Colesterol 7-alfa-Hidroxilasa/análisis , Masculino , PPAR alfa/análisis , Proteína-2 Multifuncional Peroxisomal , ARN Mensajero/análisis , Distribución Aleatoria , Ratas , Ratas Wistar
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